the journal of bombay veterinary college · 2019. 3. 22. · sangram chavan, d.s. deshmukh, amol...

The Journal of Bombay Veterinary College(An Official Orgaon of Bombay Veterinary College Alumni Association)

VOL. 21 2014 NO. 2

CONTENTS

Comparative Studies on Phytase Supplementation on Growth and 1-3Production Performance in Ducks and GeeseArchana Jain

Histomorphometrical Study of Goat Oviduct Epithelium during 4-11Luteal Phase of Estrous CycleV.D. Shankhapal, O.N. Ladukar, R.S. Dalvi, N.C. Nandeshwar and A.P. Gawande

Efficiency of Tris-egg yolk-glucose and Skim Milk Extenders for Preservation 12-16of Canine Semen at Refrigeration TemperatureBarve N.S., Gulavane S.U., Chinchkar S.R., Dagli N.R. and Kumbhar U.B.

Histological and Micrometrical Observations of Various Components of the 17-39Oviduct Before and After Extirpation of Preen Gland in LayersPatil A.D., Dhande P.L., Ranade A.S. and Ghule P.M.

Determination of Phytate and Bioavailability of Iron from Two Varieties of Legumes 40-47(Bengal gram- Cicer arietinum and Kabulichana) and Effect of Processing on themKakar D.A., Udipi S.A. and Ghugre P.S.

Histomorphological Study on Laparoscopically Excised Ovaries in Dog (Canis domesticus) 48-53Aware N.N., Shankhapal V.D., Dhande P.L., Khandekar G.S. and Kolekar V.P.

Intra-operative Evaluation of Intestinal Viability in Cases of Obstruction in Dog 54-56Jadhav P.T., S.V. Upadhye, M.S. Dhakate and B.M. Gahlod

Blood Serum Biochemical Profile of WLH Layers Before and After Uropygealectomy 57-63Patil A.D., Dhande P.L., Deshmukh B.T., Bakshi S.A., Ranade, A.S. and Ingole S.D.

Constraints of Dhangar Sheep Pastoralists’ Community in Western Maharashtra State 64-66Ajit Pokharkar, S.T. Hande, M.N. Sawant, D.N. Kale and T.R. Nikam

Effect of Supplemental Chromium on Serum Cortisol and Growth Weight in 67-72Sirohi Goat KidsS.V. Sangekar, S.D. Ingole, S.V. Bharucha and A.S. Nagvekar

Micrometrical Study on Ventricle of Heart in Sheep (Ovis aries) 73-76Kolekar V.P., Patil A.D., Dhande P.L., Shankhapal V.D., Aware N.N. and Jadhav P.M.

VOL. 21 2014 NO. 2

Constraints in Adoption of Simple but Vital, Chaff Cutter Technology for 77-81Efficient Utilization of FodderSangram Chavan, D.S. Deshmukh, Amol Patil, G.R. Gangane and Manish Sawant

SHORT COMMUNICATION

Inguinal Hernia Associated with Cryptorchidism in a Pug 82-83S.V. Upadhye, N.P. Dakshinkar, G.R. Bhojne, A.P. Gawande, G.S. Khante,A.R. Ratnaparkhi and Nitya Nirody

Retrograde Intramedullary Pinning for Repair of Humerus Fracture in a Sheep 84-85M.S. Rajhans, D.U. Lokhande, G.S. Khandekar and S.V. Gaikwad

Dystokia due to Monster with Multiple Bone Deformities in a Deoni Cow 86-87U.B. Kumbhar

Cranial Neoplasia in a Dog 88-89S.V. Upadhye, S.B. Akhare, P.T. Jadhao, G.S. Khante, Shalaka Salvekar, S.S. Pitlawarand Nitya Nirody

Mastocytoma in a Dog - A Case Report 90-91M.S. Rajhans, G.S. Khandekar , D.U. Lokhande, S.V. Gaikwad and D.J. Patil

Anaesthetic and Surgical Management of Severe Traumatic Wound on Tail in 92Indian Palm Squirrel (Funambulus palmarum)Aakash Vedpathak, K.S. Chaudhari, D.U. Lokhande, G.S. Khandekar, S.D. Tripathi,Maneesh Tiwari, Niharika Sawant and, Tenzing Sherpa

Reconstruction of Buccal Mucosa in a Dog 93-94S.V. Upadhye, S.B. Akhare, M.S. Dhakate, G.S. Khante, A.P. Gawande and Shalaka Salvekar

Surgical Management of Bilateral Mandibular Fractures in Canine using Non-locking 95-96Reconstruction platesK.S. Chaudhari, S.D. Tripathi, Aakash Vedpathak, D.U. Lokhande, G.S. Khandekar,Maneesh Tiwari and Tenzing Sherpa

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J. Bombay Vet. College (2014) 21(2)

Comparative Studies on Phytase Supplementation on Growth andProduction Performance in Ducks and Geese

Archana JainProfessor, Department of Veterinary Physiology

College of Veterinary Science & A.H., Mhow (MP) INDIA - 453446

ABSTRACT

Phytate, the salt of phytic acid, is the main storage form of phosphorus in most of theseed plants renders the phosphorus relatively unavailable to mono-gastric animals. It alsoform insoluble salts by combining with di- and tri-valant cations such as Ca, Mg, Zn, Fe, andCu, which decrease the nutrient value of feed. A protein enzyme, Phytase hydrolyzes thephytate to inositol and phosphoric acid, making these ions and phosphorus available to theanimals, which are necessary for the growth of the animals and also enhances their absorptionfrom the gut. Very scanty reports are available on the effect of phytase supplementation inducks and geese on production performance in terms of higher weight gains and feed efficiency.Three groups of six ducks each and three groups of six geese each were randomly selectedfor the present experiment. Groups I and II were kept as control, and Groups III, IV, V and VIwere supplied with Phytase @ 500 IU and 1000 IU/Kg of feed for a period of six weeks. Theresults regarding the growth were very encouraging. In ducks and geese the body weight wassignificantly higher (P<0.01) in all supplemented groups as compared to the control. TheFeed Conversion Ratio was also good in Groups III, IV, V and VI as compared to control.The body weights were higher in ducks as compared to geese. From the above study it wasconcluded that supplemented phytase highly improved the body weight gain and feedconversion efficiency in ducks and geese, so it may be beneficial to supplement phytase infeed to improve the production performance in ducks and geese.

INTRODUCTION

The application of exogenous enzymes in poultrynutrition has been driven by accruing benefits in termsof improved dietary nutrient utilization and growthperformance. Researchers have clearly demonstratedthat supplementing poultry diets with appropriateenzyme activities increases energy and N retention inchickens as well as in other species of birds includingwaterfowls. The ability of the enzyme phytase tohydrolyze phytate P and make it available forutilization by non-ruminant animals has beendemonstrated by many scientists (Perney et al., 1993;Cromwell et al., 1995; Mohammed et al., 2010). Withthe production of phytase at an affordable price,animal producers are now beginning to use phytase

to improve nutrient efficiency and reduce phosphateand N wastes from animal manure; thus reducingpotential pollution of the environment. not muchresearch has been done with phytase in the diets ofducks. In general, the P requirement for broiler chicksand ducks decreases during the later growth phasefrom 3 to 7 wk of age. Therefore, the use of microbialphytase in finisher diets for ducks should provideenough P to meet growth performance withoutsupplementing diets with inorganic P. The availabilityand utilization of phytate P released by the action ofdietary phytase can be evaluated by responses ingrowth performance The present study wasundertaken to find out the effect of phytasesupplementation to ducks and geese.

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MATERIALS AND METHODS

Three groups of six duck each and three groupsof six geese each were randomly selected for thepresent experiment. Groups I and II are kept as control,and Groups III, IV, V and VI were supplied withgrower and finisher ration along with Phytase @ 500IU and 1000 IU/Kg of feed for a period of six weeksfor ducks and geese, respectively. Ducks and geesewere reared in pens with multi nozzled waterers. Thefloors were partially covered with plastic-coated wireover a water drain. The remaining floor space wascovered with litter and included hanging cylindricalfeeders. The temperature in the room was controlledby ventilation fans and thermostatically controlled gasbrooders. Ducks and geese were provided with regularpelleted starter diet until 3 wk of age, at which timethey were individually weighed and randomly dividedand ration given as per AOAC International, (2000).A basal finisher diet containing 0.18% available P(AP) was formulated with microbial phytase addedat three levels: 0, 500, and 1,000 units/kg of diet. Thesediets did not contain supplemental P from inorganicsources. Ducks and geese were fed from 2nd to 6th wkof age. Ducks and geese had free access to feed andwater under an optimum photoperiod. Group BW andfeed intake were measured weekly to assess growthresponse.

Statistical Analysis

Data were analyzed using standard method bySnedecor and Cochran (1994).

RESULTS AND DISCUSSION

Results showing the effect of phytase on growthperformance of ducks and geese are summarized inTable 1. There was a linear increase in feed intakeand a quadratic response for weight gain due tophytase level in the diet during the last week (Week6) of the study. However, the growth performance interms of weight gain was significantly(P<0.01)higher(1,518.55, 1,615.31, 1,207.22, and 1,353.89g,respectively, in all supplemented groups than controls(996.11 and 987.22 g) but the feed intake and feedefficiency were not significantly influenced by phytasein all the supplemented groups than the control groups.

These results suggested that if ducks and geese werefed ration fortified with microbial phytase wassufficient to meet the growth requirement for P forboth ducks and geese. The phytase supplement @1000IU/ Kg of feed showed better results in both ducksand geese, though the performance in ducks is betterthan geese.

The feed conversion efficiency and the feedintake were also higher in both the species groupsfortified with phytase, but they were statistically nonsignificant than the control group.

Though very scanty reports are availableregarding the effect of phytase on productionperformance in ducks and geese, the results of thepresent study for ducks and geese were in accordancewith a variety of studies, similar improvements ingrowth performance of ducks (Orban et al., 1999;Rodehutscord et al., 2003, 2006) Cromwell et al.,(1995) in monogastrics, Yang et al. (2009) in layingducks, and Jain (2005) in poultry for weight gain, feedconversion ratio and feed intake.

CONCLUSIONS

From the above study it is concluded that theeffect of phytase supplementation in feed of duck andgeese has a very good growth promoting effect alongwith lower feed conversion ratio. Feed consumptionper Kg weight gain was also less so it may be usedfor promoting the growth of the waterfowl, but theeffect is more prominent in ducks as compared togeese. So it is suggested that the phytase should besupplemented in the diets for better performance.

REFERENCES

AOAC International (2000). Official Methods ofAnalysis. AOAC International, Arlington,

Jain, A. (2005). Effect of phytase on productionperformance in layer birds. Indian Vet. Med. J. 29:191-192.

Mohammed Kh. A., M. A. Toson, H. H. M. Hassanien,M. A. H. Solima, El-Nagar, and Sanaa H.M (2010).Effects of phytase supplementation on performanceand egg quality of laying hens fed diets containingrice bran. Egypt. Poult. Sci. 3(3): 649-659.

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Orban, J. I., O. Adeola, and R. Stroshine (1999).Microbial phytase in finisher diets of White Pekinducks: Effects on growth performance, plasmaphosphorus concentration, and leg bonecharacteristics. Poult. Sci. 78: 366–377.

Rodehutscord, M., R. Hempel, and P. Wendt (2003).Response of growing Pekin ducks to supplementationof monobasic calcium phosphate to low-phosphorusdiets. Poult. Sci. 82: 309–319.

Rodehutscord, M., R. Hempel and P. Wendt (2006).Phytase effects on the efficiency of utilisation and

blood concentrations of phosphorus and calcium inPekin ducks. Br. Poult. Sci. 47: 311–321.pmid:16787855.

Snedecor, G. W. and W. G. Cochran (1994). StatisticalMethods 8th edn., The lowa State College Press. Inc.Amer. Iowa, U.S.A .950 p.

Yang, Z. B., Z. Y. Huang, J. P. Zhou, W. R. Yang, S.Z. Jiang and G. G. Zhang (2009). Effects of a newrecombinant phytase on performance and mineralutilization of laying ducks fed phosphorus-deficientdiets. J. Appl. Poult. Res. 18(2): 284-291.

Table. 1 Effect of phytase on growth performance in ducks and geese Groups/Parameters Weight gain(g) FCR Feed

intake(kg) Ducks

Con1 996.11±31.11 2.64±0.18 2.899 Phy1 1518.56±41.15* 2.28±0.11* 3.356* Phy2 1651.31±38.59** 2.25±0.12* 3.765**

Geese Con2 987.22±35.89 2.81±0.22 2.569 Phy1 1207.22±35.89** 2.31±0.16* 2.987 Phy2 1353.89±24.02** 2.27±0.18* 3.265* * Significant difference P<0.05 ** Significant difference P<0.01

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Histomorphometrical study of goat oviduct epithelium during luteal phase of estrous cycle

V. D. Shankhapal, O. N. Ladukar, R. S. Dalvi, N. C. Nandeshwar and A. P. GawandeDepartment of Veterinary Anatomy and Histology, Nagpur Veterinary College,

Seminary Hills, Nagpur - 440 006 (M.S.)

ABSTRACT

The histomorphometrical study was conducted on thirty oviducts of non descript adulthealthy goats . The tissue pieces of 3-5 mm thickness from infundibulum, ampulla and isthmusparts of the oviducts were cut apart and preserved in 10 per cent neutral buffered formalinfor histomorphological and histochemical studies. The stained sections of infundibulum,ampulla and isthmus parts of oviduct were observed under Nikon Eclipse 80i microscope.The primary mucosal folds of all parts of oviduct were found running longitudinally andpresented secondary and tertiary branching in infundibulum and ampulla. Different types ofcells like ciliated cells, secretory cells, slender cells and rounded cells were observed in theepithelium of all parts of oviduct. Intense PAS reaction was observed at the apical border ofthe epithelial cells in infundibulum and isthmus in the luteal phase. In the tunica muscularisof isthmus, mixed Combined Ab-PAS reaction was noted for both acid as well as neutralpolysaccharides.

Keywords: Oviduct, Histomorphometry, Goat, Luteal phase

INTRODUCTIONGoat remains an important factor in the national

economy and plays a significant role in animalhusbandry sector. This is because of their highprolificacy to recover flock size, much less housingrequirement and management problems with goats.The better understanding of reproductive tract in goatis essential to achieve greater success in conceptionrate and fertilization for having higher percentage ofkidding. The problem of fertility and sterility havealways attracted considerable interest which has beenmainly directed to the reproductive tract organs likeuterus, ovaries and oviduct. The oviduct has manyroles in the process of natural conception in goat. Themajor two being retrieval of the oocyte released bythe ruptured ovarian follicle and facilitation of finalmaturation of the gametes. The oviduct is an activesecretory organ which secrets oviductal fluidsynthesized by the oviductal epithelium. The oviductalfluid provides a necessary environment for variousreproductive processes. The oviduct epithelium playsa major role in providing a suitable environment for

oocyte maturation, sperm capacitation, and earlyembryonic development (Abe,1994, Hyde andBlack,1986).

In mammals, it has been reported that theoviductal epithelium undergoes morphologicalchanges, both depending on the portion and the phaseof the oestrous cycle (Abe et al., 1999, Yániz et al.,2000). The detailed morphological knowledge ofoviduct may help in improving the treatment of itsanatomical and physiological disturbances Thehistomorphological study of oviduct in particular willhelp to correlate the importance of regional variationsin the oviduct with the fertility. This information onregional variations in the oviduct would aid in betterunderstanding of the events of reproduction in goat.Hence, the aim of the present study is to examine theregional variations in the histoarchitecture of goatoviduct in luteal phase of estrous cycle.


The histological study was conducted on thirty

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oviducts of non descript adult healthy goats whichwas carried out at the Department of VeterinaryAnatomy, Histology and Embryology, NagpurVeterinary College, Nagpur. The reproductive tractsof goat were dissected out from the carcass afterslaughter of goats at local abattoir in Nagpur. Aftercleaning and washing in normal saline, thereproductive tracts were catergorized into luteal phaseon the basis of presence of corpus luteum as suggestedby Hafez (1987).The infundibulum, ampulla andisthmus parts of oviduct were excised after trimmingthe extraneous tissue from the oviduct. The tissuepieces of 3-5 mm thickness from infundibulum,ampulla and isthmus parts of the oviducts were cutapart and preserved in 10 per cent neutral bufferedformalin for histomorphological and histochemicalstudies. After fixation of tissue for 72 hours infixatives, tissue were subjected to dehydration in theascending grades of alcohol, cleared in xylene andinfiltrated with paraffin wax as per the method ofDrury and Wallington (1980). Tissue blocks wereprepared and 3 to 5 μm thick sections were cut withrotary microtome, mounted on clean albumenizedglass slides and dried on a hot plate at 45-50°C forthree hours. Sections were stained with Hematoxylinand Eosin (HE) for general histological structure(Singh and Sulochana, 1996), Toluidine blue forhistological structure (Drury and Wallington,1980),Masson’s trichrome for collagen, elastic andsmooth muscle fibers (Bancroft and Stevens,1982),Gomoris reticulin for reticular fibers (Drury andWallington, 1980),Van Gieson’s for collagen fibers(Drury and Wallington, 1980) and Verhoeff’s methodfor elastic fibers (Drury and Wallington, 1980).

To study the histochemical status of theinfundibulum, ampulla and isthmus parts of oviduct,the paraffin sections were stained with Periodic acid-Schiff (PAS) technique for glycogen (Bancroft andStevens, 1982),Combined Alcian blue-PAS techniquefor acid and neutral mucopolysachharides (Bancroftand Stevens,1982).The stained sections ofinfundibulum, ampulla and isthmus parts of oviductwere observed under Nikon Eclipse 80i microscopeand measurements parameters viz. Number of primaryfolds , Height of primary folds, Height of epithelium,Height of Secretory cells and Height of Ciliated cellsof infundibulum, ampulla and isthmus part of oviductwere carried out. The measurements of various

parameters of three parts of oviduct thus recorded werestatistically analyzed as per the standard proceduresdescribed by Snedecor and Cochran (1980).


The mean micrometrical values for differentparameters of infundibulum, ampulla and isthmusparts of goat oviduct during luteal phase of estrouscycle are presented in Table 1.The histologicalexamination revealed three layers in the infundibulum,ampulla and isthmus parts of oviduct from withinoutward viz. tunica mucosa, tunica muscularis andtunica serosa. The lamina propria and tunicasubmucosa were blended together because of theabsence of distinct muscularis mucosa layer. Thesefindings are in agreement with the observations madeby Dellmann and Eurell (1998) and Bacha and Bacha(2000) in domestic animals. In the tunica mucosa, theprimary mucosal folds were found runninglongitudinally throughout its length in each part ofgoat oviduct and presented secondary and tertiarybranching, particularly in infundibulum and ampullain luteal phase of estrous cycle (Fig. 1) whichcorroborates with the findings of Joshi et al. (1977)in ageing goat. However, Natrajan et al. (2003a) notedabsence of tertiary folds in luteal phase in theinfundibulum of buffalo oviduct. Primary andsecondary folds in the tunica mucosa were noted bySingh and Prakash (1990) in the ampulla of goat andAbdalla (1968) in the ampulla of sheep oviduct. Abe

Fig. 1 : Photomicrograph of infundibulum showinga. Longitudinal primary mucosal foldb. Secondary mucosal foldc. Tertiary mucosal fold (HE X 100)

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and Oikawa (1991) reported finger like mucosal foldsin the fimbriae and numerous elaborate branched foldsin the ampulla whereas shorter and less branchedlongitudinal folds in the isthmus in oviduct of goldenhamster.

Presence of few primary and secondary folds andabsence of tertiary folds was noticed in the mucosaof isthmus which corroborates with the observationmade by Bloom and Fawcett (1976) in human, Leesonet al. (1985) in monkey and Bacha and Bacha (2000)in domestic animals. However Abdalla (1968) notedlow mucosal folds which did not branch in the isthmusof sheep. It may be that the variation in the findingregarding type and arrangement of mucosal folds isdue to species variation. The present observation onanastomoses of mucosal folds occupying parts oflumen in infundibulum, ampulla and isthmuscorroborates with the finding reported by Joshi et al.(1977) in the oviduct of ageing goat. The number ofthe primary folds decreased from infundibulum toisthmus. This observation is in agreement with thefindings reported by Abdalla (1968) in sheep, Joshiet al. (1977) in goat, Singh and Prakash (1990) inbuffalo, Dellaman and Eurell (1998) in domesticanimals and Natarajan et al. (2003a) in buffalo.Significant difference was also observed in the numberof primary folds between the different parts of oviductwithin luteal phase.

The observation in the present study revealedvariable pattern with regard to height of primary foldamongst the three parts of goat oviduct .In luteal phase,the height of primary fold was significantly lower inisthmus than that in infundibulum and ampulla. Thefinding of maximum height of primary fold in theinfundibulum and minimum in isthmus is in agreementwith the observation made by Joshi et al. (1977) inageing goat and Rajput and Sharma (1997) in sheep.Non-significant difference in the height of epitheliumwas revealed between infundibulum and ampulla atluteal phase, whereas it was significantly lower in theinfundibulum and ampulla than in the isthmus in lutealphase.

The epithelial lining of infundibulum andampulla in goat oviduct was found mainlypseudostratified columnar. This finding of the presentstudy is supported by the observations of Joshi et al.(1977) in ageing goat, Rajput and Sharma (1997) insheep and Dellmann and Eurell (1998) in domestic

animals. The epithelium of isthmus was mainlypsedudostratified columnar which is in line with thefindings of Joshi et al. (1977) in ageing goat andRajput and Sharma (1997) in gaddi sheep. Joshi et al.(1977) reported that towards the isthmus, theepithelium decreased in height and gradually changedto simple columnar ciliated type with no secretorycells and domes. Simple columnar type of epitheliumwas also observed by Abdalla (1968) in the sheepisthmus, Singh and Prakash (1990) in goat oviductand Steinhauer et al. (2004) in bitch oviduct. Bloomand Fawcett (1976) opined that the epithelium mightsometime appears pseudostratified when cutobliquely.

Different types of cells were observed likeciliated cell, secretory cell, slender cell and roundedcell in the epithelium of all parts of oviduct (Fig. 2).These observations are in agreement with the findingsof Abdalla (1968) in sheep, Uhrin (1983) in cow,Singh and Prakash (1990) in goats, Dellman andEurell (1998) in different domestic animals andNatrajan et al. (2003a) in buffalo. Bloom and Fawcett(1976) stated that the two types of epithelial cells ofoviduct are probably different functional status of asingle cell type. In all parts of the oviduct, the secretorycells displayed characteristic features such as theirslender shape and densely stained cytoplasm withtoluidine blue than that of ciliated cell. Prominentcytoplasmic protrusions of secretory cells were foundextended in the lumen of infundibulum and ampulla.Similar observation was noted by Natrajan et al.

Fig. 2 : Photomicrograph of infundibulum showinga. Ciliated cellb. Secretory cellc. Rounded cell (Toluidine Blue X 1000)

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(2003a) in the ampulla of buffalo oviduct, Jiwakanonet al. (2005) in sow oviduct and Tienthai et al. (2008)in buffalo oviduct. Joshi et al. (1977) reported varyingamount of budging over the surface of secretory cellsgiving dome like appearances and the nucleic andcytoplasmic fragments being shedded off theepithelium which gave it the appearance ofpseudostratified columnar type. McDaniel et al.(1968) also observed nucleated protrusion free in thelumen most commonly in the upper ampulla whichthey refer to as cellular extrusion. Tienthai et al. (2008)stated that cytoplasmic protrusion of secretory cellsmight be a part of the process in which dead epithelialcells are eliminated. No cytoplasmic protrusion wasnoticed in the secretory cells of isthmus epitheliumwhich corroborates with the finding of McDaniel etal. (1968) in bovine oviduct and Steinhauer et al.(2004) in bitch. The observation of numeroussecretory cells in infundibulum and ampulla than inisthmus confirm the findings of Abdalla (1968) insheep and Joshi et al. (1977) in ageing goat. Thisfinding of the present study indicates more secretoryactivity in the infundibulum and ampulla parts ofoviduct in goat. Bloom and Fawcett (1976) reportedthat the secretion of the secretory cell may providethe ovum with nutritive material in some species.

The cytoplasm of ciliated cells was found lightlystained than that of secretory cells in all parts ofoviduct of goat. Numerous ciliated cells wereobserved than secretory cells in the epithelium of allparts of oviduct which corroborates with the findingof Rajput and Sharma (1997) in sheep. The ciliatedcells of isthmic epithelium were elongated in shapewhich corroborates with the finding of McDaniel etal. (1968) in bovine oviduct. They further noticed thatciliated cells in the ampulla varied in size from large,lightly stained, vacuolated cells to intensely stainedpeg cells. The height of ciliated cells of infundibulum,ampulla and isthmus parts of oviduct variedsignificantly. The height of ciliated cells wassignificantly more in the isthmus than in infundibulumand ampulla which might be due to increase in thecytoplasmic matrix of ciliated cells of isthmus.Contrary to this, Natarajan et al. (2003b) noted thatcell height declined gradually from infundibulum toisthmus in all reproductive phases in buffalo oviduct.Abe et al. (1999) revealed that the height of epithelialcells was reduced in all regions during the luteal phase

and this reduction of height was most dramatic in theciliated cells in the fimbriae and ampulla. They furthersuggested that the reduction in the height of ciliatedcells is responsible for the reduction in the number ofcilia on the surface of the goat oviductal epitheliumat the luteal phase. Significant difference in the heightof secretory cells was found amongst infundibulum,ampulla and isthmus at luteal phase. It may be thatthe high level of circulating progesterone hormone inluteal phase of estrous cycle reduces the secretoryactivity of secretory cells, which support the statementof McDaniel et al. (1968) reported that thecytoplasmic content of ciliated and secretory cells inboth the ampulla and the isthmus was increased inthe presence of estrogen and decreased in the presenceof progesterone. Leese et al. (2001) suggested thatchange in the cell morphology like hypertrophy,maturation and cell height indicate changes insynthetic activity that contributes to the variations inthe oviductal fluid production. The finding ofreduction in the height of the ciliated and secretorycells of all parts of oviduct in goat during luteal phaseof estrous cycle is in line with the observationsrecorded by Tienthai et al. (2008) in buffalo oviduct.They further reported that the changes of epithelialcell height found in buffalo oviduct could be mainlydependent on the presence of ciliated cells (ciliation)at follicular phase and loss of ciliated cells (deciliation)at the luteal phase especially in infundibulum andampulla .They suggested that regular cycle ofciliogenesis and deciliation by epithelial cells ofbuffalo oviduct depends on estrous cycle and reflectfunction in different regions.

In between ciliated and secretory cells, rareoccurrence of slender cells noticed in each part ofoviduct. The slender shape of these cells might bedue to release of secretory material from secretorycells. Similar cells were reported as ‘Peg Cells’ byJoshi et al. (1977) in ageing goat and Hafez (1987)in farm animals. Joshi et al. (1977) stated that thesecells might be representing the degenerated form ofnon-secretory cells. They further opined that thesecells might be in intermediate phases of transformationof secretory to non-secretory cells and vice versa.Whereas, Hafez (1987) stated that peg cells werepresumably depleted secretory cells. Hadek (1955)reported similar cells in sheep during metestrus andnamed as ‘Rod Cells’. They considered that these cells

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are derived from the extrusion of their nucleipresenting a degenerated form of the non-ciliated cells.McDaniel et al. (1968) reported intensely stained pegcells in the ampulla of bovine oviduct which did notpossess cilia in isthmus. Samuelson (2007) describedthat as oocytes are released during ovulation; the non-ciliated columnar cells become metabolically moreactive and typically increase in their height whichbeing sometimes referred to as peg cell. Verhage etal. (1973) opined that these peg cells appeared to be astage in the dedifferentiation of the secretory andciliated cells and therefore are not a specific cell type.The finding of rounded cells like lymphocytes andneutrophils were noticed at the base of epithelium ineach part of goat oviduct confirms the observationsrecorded by McDaniel et al. (1968) in cow oviduct,Joshi et al. (1977) in the oviduct of ageing goat andTienthai et al. (2008) in buffalo oviduct. Such cellswith a halo around noticed in the sow oviduct termedas ‘Clear Cell’ by Bal and Getty (1970) in sow oviduct.They further concluded that these cells within theepithelium and in the lamina propria along with theplasma cells may not only be performing trophicfunction, but could also be a readily available sourceof immunogenic response. Ellis et al. (1986) statedthat the oviduct of domestic animals has a mucosalsystem which is required for maintaining an asepticintraluminal fluid. Tienthai et al. (2008) opined thatthe presence of intraepithelial lymphocytes in thebuffalo oviduct might involve in the regulation of localimmune response and production of antigen specificantibodies. In isthmus, few rounded cells were noticedin the epithelium as compared to infundibulum andampulla. This finding of the present study support theobservation made by Jiwakanon et al.(2005) whonoted higher number of lymphocytes in thesubepithelial connective tissue of the infundibulumand ampulla than that in the isthmus of sow oviduct.They stated that the immune cell reaction had to below in the isthmus because both spermatozoa as storedbefore fertilization and the semi allogenic conceptuscould survive.

In all parts of goat oviduct, the lamina propriawas found merged with submucosa constitutingpropria-submucosa as no muscularis mucosae waspresent. This layer was made up of loose connectivetissue consisting dense collagen as well as reticularfibres (Fig. 3). Similar observations were recorded

by Dellman and Eurrell (1998) in domestic animals,Abdalla (1968) in sheep and Rajput and Sharma(1997) in sheep.Scanty infiltrated cells likelymphocytes, fibroblast and plasma cells wereobserved in the lamina propria of infundibulum,ampulla and isthmus(Fig. 4). Similar finding wasnoted by Natrajan et al. (2003a) in buffalo oviduct.DuBois et al. (1980) observed mast cells in the laminapropria in the mucosa of ampulla and isthmus ofoviduct while they observed lymphocytes to migratethrough the uterine tube epithelium in cow. Theyfurther opined that histamine produced by mast cellsmight play a role in the inhibition of cytotoxic activityof lymphocytes in the oviduct during estrus. Thepresent observation on the occurrence of few smooth

Fig. 3 : Photomicrograph of ampulla showing network ofcollagen fibers (arrowhead) (Van Gieson’s X 200)

Fig. 4 : Photomicrograph of infundibulum showinga. Lymphocyteb. Plasma cell c.Fibroblast in the stroma of mucosal fold(Toluidine Blue X 1000)

9


muscle cells in the propria-submucosa of each part ofoviduct is in line with the finding of Joshi et al. (1977)in ageing goat. These smooth muscle cells along withcollagen and reticular fibers may play role in oviductalmotility for transportation of gametes.

The observation regarding presence of innercircularly and outer longitudinally arranged bundlesof smooth muscle fibers in the tunica muscularis ofthree segments of goat oviduct is in accordance withthe findings reported by Rajput and Sharma (1997)in sheep and Lesson et al. (1985) in monkey. Whilecontrary to this, Singh and Prakash (1990) reportedorientation of few bundles of smooth muscle fibersin an oblique or longitudinal direction on the peripherybut no distinct outer longitudinal layer in the tunicamuscularis in goat, which believed to play animportant role in the transport of ovum by theircontractions in goat. Halbert et al. (1989) opined thatthe oscillation may serve as an additional mechanismof transport as cilia clearly provide sufficientpropulsive force to transport egg masses to the site offertilization despite the absence of muscular activityin rat oviduct. They further opined that undulatorymotion caused by oviductal muscle may facilitateremoval of the cumulus mass that surrounds the eggas the decrease in size may promote entry into thenarrower isthmic portion of oviduct and oscillationmay enhance the likelihood of successful contactbetween ova and sperm. Despite these possibilities,they suggested that muscle contractility in the ampullaof rat oviduct is not necessary for fertility. Dellmannand Eurell (1998) reported that the tunica muscularisconsisted chiefly of circular smooth muscle bundles,but isolated longitudinal and oblique bundles alsooccurred which gave off radial strands into themucosa. They stated that in the ampulla, ciliary activityis the primary force propelling the egg towards theisthmus, but in some species, muscle contractility isalso involved. The finding of thick and prominentmuscle layer in isthmus of goat oviduct corroborateswith observation of Dellmann and Eurell (1998) andBacha and Bacha (2000) in domestic animals.Dellmann and Eurell (1998) stated that in isthmusmuscle contractility is the primary force propellingthe zygote towards the uterus with ciliary activityinvolved in some species.

The tunica serosa was found to be made up ofloose connective tissue consisting of collage fibers

and few reticular as well as elastic fibers which waslined by flattened mesothelial cells. Similar findingswere noted by Singh and Prakash (1990) in goat andRajput and Sharma (1997) in sheep.

The finding of PAS positive reaction in theepithelium of all parts of goat oviduct is incorroboration with the observations reported byMcDaniel et al. (1968) in cow, Singh and Prakash(1990) in the ampulla of goat oviduct, Rajput andSharma (1997) in sheep and Gregoraszczuk et al.(2000) in pig. Steinbach and Smidt (1970) observedmaximum of PAS positive material during latefollicular phase and minimal of PAS positive materialduring the late luteal phase in the epithelium of sowoviduct. The presence of PAS positive material in theepithelium of goat oviduct may be as a source fortransportation of gametes, sperm maturation,fertilization and early embryo transport. Intense PASreaction was observed at the apical border of theepithelial cells in infundibulum and isthmus in theluteal phase which support the observation made byAbdalla (1968) in sheep oviduct. Gadegone et al.(1981) observed that glycogen in ciliated cells ofIndian goat oviduct undergoes no variation duringdifferent phases of estrous cycle and the intensity ofPAS reaction was less in isthmus region of oviduct.Gregoraszczuk et al. (2000) opined that glycogen inthe preovulatory stage might be functioning as asource of energy for ciliary movement and as anourishment for ovum. Abdalla (1968) notedsupranuclear PAS positive material inside the cellsduring proestrous and at the border of the epithelialcells in the cytoplasmic projections by late proestrousand the onset of metestrous. The secretory cellsshowed strong PAS positive activity in thesupranuclear cytoplasm and in the secretory bleb ofthe cells in the infundibulum and ampulla whereas inisthmus the secretory cells showed weak activity forPAS positive material. This finding corroborates withthe finding of Joshi et al. (1977) who also observedstrongly PAS positive secretory material in the goatoviduct, which reduced towards the isthmus. Theyreported intracellular PAS positive secretory materialat estrus and metestrus, which was found released intolumen in the early diestrus. Moderate to strong PASreaction was observed in the tunica muscularis forpresence of glycogen in infundibulum and isthmus.Hafez (1987) also reported higher amount of glycogen

10


in the oviductal musculature after ovulation. Heopined that the varying patterns of oviductalcontraction may be associated with cyclic changes inglycogen content of oviductal musculature.

Intense and weak combined Ab-PAS reactionwas noticed for presence of neutral and acidmucopolysaccharides respectively in the apicalepithelial border in infundibulum and ampulla.Contrary to this, Singh and Prakash (1990) noticedonly acid mucopolysaccharides in ampulla ofgoat,while Natarajan et al. (2003a) reported presenceof sulphated acid mucopolysaccharides in the oviductof buffalo. In isthmus, moderate combined Ab -PASreaction was observed for presence of acid as well asneutral mucopolysaccharides. Gregoraszczuk et al.(2000) noted that polysaccharides granules wereminimal or absent in the isthmus during midlutealphase in pig. They further reported that numerousgranules of polysaccharides aggregated in the isthmusepithelium after ovulation in pig could be the majorenergy source for embryogenesis when embryo travelsdown the oviduct during the early cleavage stage. Thefinding of moderate to strong AB-PAS reaction forthe presence of acid mucopolysaccharides in thelamina propria in each part of oviduct in both phasesof estrous cycle is in line with the observation madeby Joshi et al. (1977) in the oviduct of ageing goat.In the tunica muscularis of infundibulum and ampulla,moderate to strong Combined Ab-PAS activity wasnoted for neutral mucopolysaccharides. In the tunicamuscularis of isthmus, mixed Combined Ab-PASreaction was noted for both acid as well as neutralpolysaccharides (Fig. 5) . In the tunica serosa of

infundibulum and isthmus, strong Combined Ab-PASactivity was noted for neutral mucoploysaccharides.

REFERENCES

Abdalla, O. (1968). J. Anat., 102(2): 333-344.

Abe, H. (1994). Anat. Rec., 240: 77-85.

Abe, H. and T. Oikawa (1991). J. Anat., 175: 147–158.

Abe, H.; Onodera, M.; Sugawara, S.; Satoh, T. andH. Hoshi, (1999). J. Anat., 195(4): 515-521.

Bacha, William J. and Jr. Linda M. Bacha (2000).

Table 1. Mean micrometrical values for different parameters of infundibulum, ampulla and isthmus parts of goat oviduct during luteal phase of estrous cycle Sr No.

Parameters Part of Oviduct Infundibulum Ampulla Isthmus

1 Number of primary folds** 38.83c ± 0.58 25.00b ± 0.59 16.83a ± 0.35

2 Height of primary folds** (µm) 385.33b ± 22.81 354.90b ± 21.64 168.30a ± 8.87

3 Height of epithelium** (µm) 19.66a ± 0.51 19.55a ± 0.65 25.21b ± 0.73

4 Height of ciliated cells** (µm) 17.81a ± 0.38 18.19a ± 0.50 25.23b ± 0.60

5 Height of secretary cells** (µm) 16.47a ± 0.59 19.97b ± 0.42 22.55c ± 0.63

Similar superscripts within the respective row indicates non significant differences, ** P<0.01

Fig. 5 : Photomicrograph of isthmus showing combinedAB-PAS reaction for acid and neutral mucopolysaccharidesa. Tunica Mucosab. Lamina propria-submucosac. Tunica muscularis (Combined AB-PAS X 200)

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Color Atlas of Veterinary Histology, 2nd Edn.,Lippincott Williams and Wilkins, A Wolters KluwarCompany, Maryland. p.p. 221-229.

Bal, H.S. and R. Getty (1970). Cited by Joshi et al.(1977).

Bancroft, J.D. and A. Stevens (1982). Theory andPractice of Histological Techniques, 2nd Edn., ChurchilLivingstone, Edinburgh, London, Melbourne and NewYork, pp. 110 - 114.

Bloom, William and Don W. Fawcett (1976). ATextbook of Histology 10th Edn., W. B. Saunders Co.,Philadelphia. pp. 881– 883.

Dellmann, H.D. and J. Eurell (1998). Textbook ofVeterinary Histology, 5th Edn., Lippincott Williamsand Wilkins, A Wolters lower Co. Maryland, pp. 255-257.

Drury , R.A.B. and E.A. Wallington (1980). Carleton’sHistological Technique,5th Edn., Oxford UniversityPress, New York, pp. 36-198.

Dubois, J.A.; R. J. Wordinger and J. F. Dickey (1980).Am. J. Vet. Res., 41 (5): 806 -809.

Ellis, W.A.; P.J. McParland; D.G. Bryson; A.B.Thiermann and J. Montgomery (1986). Cited byTenthai et al. (2008).

Gadegone, M.M., Sapkal V.M., and Sawhney, G.(1981). Zeitschrift fur mikroskopisch-anatomischeForschung, 95 (2): 230-8.

Gregoraszczuk, E.L., Cala M. and A.E. Witkowsk,(2000).Folia Biologica, 48 (3-4): 85-90.

Hadek, R. (1955). Cited by Joshi et al. (1977).

Hafez, E.S.E. (1987). Reproduction in Farm Animals,4th Edn., K.M. Varghese Co., Bombay, pp. 17-167.

Halbert, S.A., D.R. Becker and S.E. Szal (1989). Biol.Reprod., 40(6):1131-6.

Hyde, B.A. and Black, D.L. (1986). J. Reprod. Fert.78: 83-91.

Jiwakanon, J.; E. Persson, K. Kaeoket and A.M. Dalin(2005). Reproduction in Domestic Animals,

40(1): 28 – 39.

Joshi, C.L.; B.S. Nanda and R.P. Saigal (1977). Anat.Anz., 141(2): 163-73.

Leeson, C.R.; T.S. Leeson and A.A. Paparov (1985).Textbook of Histology” 5th Edn., W.B. Saunders Co.,Philadelphia, pp. 470-472.

Leese H.J., J.I. Tay, J. Reischl and S.J. Downing(2001). Reproduction, 121: 339-346.

McDaniel, H.W.; H. Scalzi and D.L. Black (1968). J.Dairy Sci., 51(5): 554 – 561.

Natrajan, T.; R.V. Prasad; K. Kakade and K.V. Jamuna(2003a). Ind. J. Anim. Sci., 73(5): 484-487.

Natrajan, T.; R.V. Prasad; K. Kakade and K.V. Jamuna(2003b). Ind. J. Anim. Sci., 73(5): 519-521.

Rajput, R. and D.N. Sharma (1997). Ind. Vet. J., 74:580-583.

Samuelson, Don A. (2007). Textbook of VeterinaryHistology, Saunders Elsevier, Missouri pp. 457-459.

Singh, G. K. and P. Prakash (1990). Indian Vet. J., 67:152 -154.

Singh, U.B. and S. Sulochana (1996). Handbook ofHistological and Histochemical Technique .1st Edn.,Premier Publishing House, Hyerabad, pp. 20-94.

Snedecor, G.W. and Cochran, W.G. (1980). StatisticalMethods. 6th Edn., Oxford and IBH Publishing House,Calcutta.

Steinbach J. and D. Smidt (1970). J. Anim. Sci., 30:573-577.

Steinhauer, N.; A. Boos and A.R. Gunzel-Apel (2004).Reprod. Domest. Anim. 39 (2):110-119.

Tienthai, P.; K. Sajjarengpong and M .Techakumphu(2008). Thai J. Vet. Med., 38(1): 27-38.

Verhage H.G., J.H. Jr Abel, W.J. Jr Tietz and M.D.Barrau (1973). Biol. Reprod., 9 (5): 460-74.

Yániz, J.L., López-Gatius, F., Santolaria, P. & JuneMullins, K. (2000). Anat. Rec., 260(3):268-78.

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Efficiency of tris-egg yolk-glucose and skim milk extenders for preservation of canine semenat refrigeration temperature

Barve N.S., Gulavane S.U., Chinchkar S.R., Dagli N.R. and Kumbhar U.B.Department of Animal Reproduction, Gynaecology and Obstetrics,

Bombay Veterinary College, Parel, Mumbai - 400 012.

ABSTRACT

Semen was collected from six large breed dogs twice at weekly interval. After collectionsemen was evaluated by macroscopic, microscopic and sperm function tests. Then semenwas dived in to two aliquots which were diluted with Tris egg yolk Glucose (TEYG) andSkimmed milk extenders (SME). Diluted semen was transported in thermos flask at 50C within40 min to one hrs to the laboratory for preservation at refrigeration temperature up to 24hours. Preserved semen was also evaluated at 0 and 24 hours of preservation with microscopicand sperm function tests. After 24 hours of preservation sperm motility, live sperm, HOSpositive sperm and intact acrosome sperm percentage dropped significantly in both dilutors;however motility and live sperm count were above 50 percent in TEYG. Also at 24 hoursintact acrosome sperm percent was significantly higher and abnormal sperm count wassignificantly lower in TEYG as compared to SME. It was concluded that TEYG was a betterdilutor for preservation of canine semen up to 24 hours at refrigeration temperature ascompared to SME.

Keywords: Tris egg yolk glucose, Skimmed milk extenders, Canine semen

INTRODUCTION

With progress of industrialization fromagriculture, change in social life from joint family tonuclear family, there is increased trend of keeping dogsas companion animal. There are some owners whowant to breed their pets so that their pets can have afamily and pet’s name will be carried forward intothe future. Sometimes, natural mating fails due tovarious factors involving both the male and the femaledogs. Due to failure of natural mating, many ownersand breeders want to go ahead with artificialinsemination (AI). In such cases, semen preservationfor short duration and artificial insemination isnecessary. Many a times the Veterinarians has tocollect canine semen at a place away from AIlaboratory like owners place or a clinic and itspreservation is a must to conserve the sperm motilitytill AI. Therefore, it was thought worthwhile tocompare preservation of canine semen at refrigeration

temperature (5°C) by TRIS-egg yolk-glucose andskim milk extenders carrying out the semen evaluationto predict the semen quality by evaluatingmacroscopic, microscopic characteristics and spermfunction tests after collection at owners’ place andtransported in dilutor at refrigeration temperature(5°C).


In present study six dogs of large breeds,weighing more than 20 kg were selected for the semencollection. Semen was collected twice aseptically andhygienically at an interval of one week from all sixdogs by digital manipulation method. Semen wascollected at private clinics or owners’ places, dilutedand then transported to the Department of ARGO,Bombay Veterinary College, within 40 minutes to 1hour, in 5°C water bath with ice pack in thermos flask.Immediately after collection, semen samples were

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evaluated for macroscopic tests, viz. volume, colour,consistency and density and microscopic tests, viz.mass activity, initial sperm motility, total spermconcentration, live sperm count and abnormal spermcount and sperm function tests viz. acrosomeintactness test derived by Giemsa staining and GelatinDigestion Test and Hypo-osmotic test (HOST). Semensamples were divided into two aliquots which werethen diluted in TRIS-Egg yolk-glucose extender(Group-I) and skim milk extender (Group-II). Semendilution rate was volume : volume. Immediately aftertransportation, initial sperm motility was observed toconfirm survival of sperm and the diluted sample wasplaced under refrigeration in water bath. Then thediluted semen were analysed at 24 hours formicroscopic and sperm function tests. The resultsobtained were then analysed as per Snedecor andCochran (1994).

RESULTS AND DISCUSSIONS

Six dogs were subjected to semen evaluationtwice with the minimum interval of one week. It wasobserved (table 1) that the volume of sperm-richfraction of neat semen as 271±0.21 ml with the rangeof 2.00 to 4.00 ml, which was thick, had density ofDDD and milky in colour in dogs above 20 kg bodyweight. The results also showed initial sperm motilitypercentage, live sperm percentage, percentage ofmorphologically abnormal spermatozoa and totalsperm concentration as 70.83 ± 3.79, 71.58 ± 3.24,10.67 ± 0.38 and 162.20 ± 6.26 millions/ml,respectively in neat semen. The result of the sperm

having intact plasma membrane percentage judgedby HOST, percentage of sperm having intact acrosomederived from Gelatin Digestion Test and Giemsastaining were 89.17 ± 1.32, 85.17 ± 0.69 and 91.83 ±0.72, respectively in neat semen. All the aboveparameters were in the normal range of neat caninesemen.

Semen was diluted in two extenders TRIS-Eggyolk-glucose extender and Skimmed milk extender.It was evaluated and the results of the same arepresented in table no.2.

From table 2 it was observed that at 24 hours,initial sperm motility percentage dropped from70.83±3.79 to 52.50 ± 5.12 and 48.33 ± 6.49 in TRIS-Egg yolk-glucose dilutor and in skim milk dilutorrespectively. Though statistically difference was notsignificant for both groups, initial motility was above50 percent in TRIS-Egg yolk glucose dilutor whichindicates that TRIS was better dilutor for semenpreservation compared to skim milk dilutor. Similarly,Rota et al. (1995) reported that egg yolk-TRISextender was superior to egg yolk-milk dilutor forpreservation of semen at 4°C. Similar observationson semen stored in TRIS-Egg yolk-glucose extenderwere made by Gill et al. (1970), Daiwadnya (1987)and Pinto et al. (1999) for 24 hours preservation. At24 hours, Province et al. (1984) reported spermmotility of 25 percent in TRIS dilutor and 15 percentin skim milk dilutor, Bouchard et al. (1990) reportedsperm motility of 39 percent in TRIS-Egg yolk-glucose dilutor, which is lower than the observationof the present study. However, higher observationswere reported by Rota et al. (1995), Rijsselaere et al.

Table 1 The pooled data regarding Macroscopic, microscopic and sperm function test of neat canine semen.

Sr.No Parameters Observations (n=06)

2 Volume of sperm rich fraction (ml) 2.71± 0.21 3 Colour Milky 4 Consistency Thick 5 Density DDD 6 Initial sperm motility percent 70.83±3.79 7 Live sperm percentage percent 71.58±3.24 8 Abnormal spermatozoa percent 10.67±0.38 9 Sperm concentration (million/ml) 162.20±6.26

10 HOS positive sperm percent 89.17±1.32 11 Intact acrosome sperm percent (Gelatin Digestion) 85.17±0.69 12 Intact acrosome sperm percent (Giemsa staning) 91.83±0.72

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(2002), Tsutsui et al. (2003), Ponglowhapan et al.(2004), Verstegan et al. (2005), Shahiduzzaman andLinde-Forsberg (2007), Michael et al. (2009), Kmentaet al. (2011), Bencharif et al. (2013) and Hori et al.(2014) in TRIS-Egg yolk-glucose dilutor. Similarobservations were made by Romagnoli (2002) forskim milk dilutor. Observations made by Bouchardet al. (1990), Rota et al. (1995) and Sánchez et al.(2006) were higher than the observations made forskim milk in the present study at 24 hours.

From table 2 it was noted that at 24 hours, livesperm percentage dropped from 71.58 ± 3.24 to 51.83± 4.13 and 48.83 ± 4.11 in TRIS-Egg yolk-glucosedilutor and in skim milk dilutor respectively. Thoughlive sperm percentage was slightly higher in TRIS-Egg yolk-glucose dilutor at 24 hours compared to skimmilk, it renders the sample unsuitable for artificialinsemination beyond 24 hours. Similar observationwas made by Daiwadnya (1987) and Ponglowhapanet al. (2004) for TRIS-Egg yolk-glucose dilutor.However, higher live percentage for TRIS-Egg yolk-glucose dilutor at 24 hours were reported by Kumi-Diaka and Badtram (1994), Rijsselaere et al. (2002),Tsutsui et al. (2003), Michael et al. (2009) and Kmentaet al. (2011). Observations for skim milk dilutor at 24hours in the present study were more than those madeby Sánchez et al. (2006).

From table 2 it was noted that at 24 hours, spermabnormality percentage significantly increased from10.76±0.38 to 12.50 ± 0.31 and 13.58 ± 0.29 in TRIS-Egg yolk-glucose dilutor and in skim milk dilutorrespectively. Similar observations were made by

Daiwadnya (1987), Rijsselaere et al. (2002) andMichael et al. (2009) for TRIS-Egg yolk-glucosedilutor. At 24 hours, Tsutsui et al. (2003) reported 8percent abnormal sperm of for TRIS-Egg yolk-glucosedilutor which was lower than that observed in presentstudy. Abnormal sperm of 28 percent was reportedby Kmenta et al. (2011) for TRIS-Egg yolk-glucosedilutor at 24 hours which was higher than theobservation of present study. For skim milk dilutor,the observations reported in the present study werelower than those reported by Sánchez et al. (2006) at24 hours.

From table 2 it was noted that at 24 hours, intactplasma membrane percentage significantly droppedfrom 89.17±1.32 to 85.25 ± 1.50 and 86.00 ± 1.50 inTRIS-Egg yolk-glucose dilutor and in skim milkdilutor respectively. Similar observation was made byRota et al. (1995) for TRIS-Egg yolk-glucose dilutor.However, lower intact plasma membrane percentagein TRIS-Egg yolk-glucose dilutor at 24 hours wereobserved by Ponglowhapan et al. (2004), Michael etal. (2009), Treulen et al. (2012) and Bencharif et al.(2013) for TRIS-Egg yolk dilutor. Similarobservations were made by Rota et al. (1995) for skimmilk dilutor. At 24 hours, Sánchez et al. (2006)observed 66.8 percent intact sperm plasma membranewhich was lower than that observed in the presentstudy for skim milk dilutor.

From table 2 it was noted that at 24 hours, intactacrosome percentages derived by Giemsa stainingsignificantly dropped from 91.83±0.72 to 89.83 ± 0.87and 88.92 ± 0.92 in TRIS-Egg yolk-glucose dilutor

Table no 2 Evaluation of semen extended in different dilutor

Time interval 0 hours 24 hour Parameters TRIS-Egg yolk-

glucose extender Skim milk extender

Sperm motility percentage 70.83±3.79a 52.50±5.12b 48.33±6.49b Live sperm count 71.58±3.24a 51.83±4.13b 48.83±4.11b

Abnormal sperm count 10.67±0.38 c 12.50±0.31b 13.58±0.29a HOS positive sperm percent 89.17±1.32a 85.25±1.50b 86.00±1.50b

Intact acrosome sperm percent (Giemsa staning)

91.83±0.72a 89.83±0.87b 88.92±0.92c

Intact acrosome sperm percent (Gelatin digestion test)

85.17±0.69a 83.42±0.70b 82.42±0.78c

In a raw the mean with common super script do not differ significantly

15


and in skim milk dilutor respectively. Similarobservation was made by Ponglowhapan et al. (2004)for TRIS-Egg yolk-glucose dilutor. At 24 hours, Rotaet al. (1995), Michael et al. (2009), Treulen et al.(2012) and Bencharif et al. (2013) reported lowerintact acrosome percentage in TRIS-Egg yolk-glucosedilutor than observed in present study. At 24 hours,Iguer-Ouada and Verstegen (2001) and Rijsselaere etal. (2002) observed intact acrosome percentage ofTRIS-Egg yolk-glucose dilutor which was higher thanthose observed in the present study. Lower intactacrosome percentage than that observed in the presentstudy for skim milk dilutor was reported by Rota etal. (1995) as 73% at 24 hours of preservation.

From table 2 it was noted that at 24 hours, intactacrosome percentages derived by Gelatin digestiontest significantly dropped from 85.17±0.69 to83.42±0.70 and 82.42±0.78 in TRIS-Egg yolk-glucose dilutor and in skim milk dilutor respectively.The data showed that intact acrosome percentage wassignificantly higher in TEYG as compared to SMEdilutor at 24 hours of preservation. This clearly statesthat TEYG preserves the acrosomes better than SME.When available literature was scanned, there was noreferences found to compare the acrosomal intactnessby Gelatin Digestion Test for canine semen post-preservation at 24, hours.

CONCLUSIONS

Though canine semen can be preserved in bothTRIS-Egg yolk-glucose and skim milk dilutor up to24 hours at refrigeration temperature; TRIS-Egg yolk-glucose dilutor is the better dilutor of the two forpreserving semen as it maintained initial motilityabove 50 % and live percentage above 50 %.,preserves intact acrosome and delays the degenerationof the sperm.

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Gill H.P., C.F. Kaufman, R.H. Foote and R.W. Kirk(1970). Artificial insemination of beagle bitches withfreshly collected, liquid-stored and frozen-storedsemen. American Journal of Veterinary Research 31:1807-1813.

Hori T., R. Yoshikuni, M. Kobayashi and E.Kawakami (2014). Effects of Storage Temperature andSemen Extender on Stored Canine Semen. J. Vet. Med.Sci. 76(2): 259-263.

Iguer-Ouada M., J. Verstegen (2001). Long-termpreservation of chilled canine semen: effect ofcommercial and laboratory prepared extenders.Theriogenology 55: 671-684.

Kmenta I., C. Strohmayer, F. Müller-Schlösser, S.Schäfer-Somi (2011). Effects of a lecithin and catalasecontaining semen extender and a second dilution withdifferent enhancing buffers on the quality of cold-stored canine spermatozoa. Theriogenology 75: 1095-1103.

Kumi-Diaka J. and Badtram G. (1994). Effect ofstorage on sperm membrane integrity and otherfunctional characteristics of canine spermatozoa: in-vitro bioassay for canine semen. Theriogenology 42:815-829.

Michael A.J., C. Alexopoulos, E.A. Pontiki, D.J.Hadjipavlou-Litina, P. Saratsis, H.N. Ververidis, C.M. Boscos (2009). Effect of antioxidantsupplementation in semen extenders on semen qualityand reactive oxygen species of chilled caninespermatozoa. Animal Reproduction Science 112: 119-135.

Pinto C., D. Paccamonti, B. Eilts (1999). Fertility inbitches artificially inseminated with extended, chilledsemen. Theriogenology 52(4): 609-616.

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Province C.A., R.P. Amann, B.W. Pickett, E.L. Squires(1984). Extenders for preservation of canine andequine spermatozoa at 5°C. Theriogenology 22(4):409-415.

Rijsselaere T., A. Soom, D.Maes et al. (2002). Effectof centrifugation on in-vitro survival of fresh dilutedcanine spermatozoa. Theriogenology 57 (6): 1669-1681.

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Rota A., B. Ström, C. Linde-Forsberg (1995). Effectsof seminal plasma and three extenders on caninesemen stored at 4°C. Theriogenology 44(6): 885-900.

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Snecedor G.W and Cochran W.G (1994). Statisticalmethods, 8th Edn. Affiliated East west press: 491.

Treulen F., R. Sanchez and J. Risopatrón (2012).Effects of seminal fluid fractions on plasma andacrosome membrane integrity and mitochondrialmembrane potential determined by flow cytometryin chilled canine spermatozoa. Reproduction inDomestic Animals 47 (6): 1043-1048.

Tsutsui T., T. Tezuka, Y. Mikasa, H. Sugisawa, N.Kirihara, T. Hori and E. Kawakami (2003). Artificialinsemination with Canine Semen Stored at a LowTemperature. J. Vet. Med. Sci. 65(3): 307-312.

Verstegen J., K. Onclin and M. Iguer-Ouada (2005).Long-term motility and fertility conservation ofchilled canine semen using egg yolk added Tris-glucose extender: in-vitro and in-vivo studies.Theriogenology 64: 720-733.

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Histological and micrometrical observations of various components of the oviduct before andafter extirpation of preen gland in layers.

Patil, A.D., Dhande, P.L., Ranade, A.S. and Ghule, P.M.Department of Veterinary Anatomy and Histology,


ABSTRACT

After extirpation of preen gland, histomorphological and micrometrical alterations inthe oviduct of young and spent hens were studied. Young hens aged 32 weeks (50 birds) andspent hens of 90 weeks (50 birds) were subdivided into three groups each consisting of startcontrol group (10 birds), end control group (20 birds) and preen gland extirpated group (20birds). Ten birds from each group were sacrificed. Histomorphological and the micrometricalchanges studied during the research work.

The numbers of mucosal fold, height of surface epithelium, goblet cells & thickness oftunica mucosa were increased from Group I to III and from IV to VI. The tunica submucosaof infundibulum, magnum and uterus increased in thickness and was thickest in preen glandextirpated Groups III and VI. The muscular layer was initially thin which increased in thicknessfrom Group I to III in young hens and from Group IV to VI in spent hens. The tunica serosawas thin in Group I and IV, which increased and found significantly thicker in Group III andVI.

Keywords: Preen gland, Extirpation, uropygealectomy, Hens, Oviduct, Tunica, Tubular glandand Histology.

INTRODUCTION

Preen or uropygeal gland is only one pear shapedholocrine, cutaneous, oil secreting gland situated atthe base of tail or coccygeal vertebra which is welldeveloped in aquatic birds. It is used in oiling thefeathers, which protect feathers from getting wet andis antifungal and antibacterial in activity. Birds spreadthis secretion all over the body by the process ofpreening which help in attracting the male and alsovitamin D synthesis. Extirpation of the uropygeal hasslight effect on increase in the weight gain (Ray etal., 1971).

France investigator Giraudeau (2010) establisheda new link between important avian self-maintenancebehavior and aspects of maternal health andreproduction. They suggested that higher yolkcarotenoid levels in eggs laid by preen-gland-restricted females may serve to boost health of

offspring that would hatch in a comparativelymicrobe-rich environment.

In male fowl, the effect of uropygealectomy wasstudied on physiological and reproductive traits ofbroiler breeder males. There is good evidence that,uropygealectomy could be used for support fertilityin broiler breeder males which suffer from age relateddecline in fertility (Ismail, 2005).

Though the histomorphology and the functionof the avian oviduct have been much described fordomestic hen in the literature, no work correlatingeggs production and histomorphology of the hen’soviduct after removal / extirpation of preen gland(uropygealectomy) were traced. The presentinvestigation is, therefore, aimed at the study ofhistomorphological and micrometrical aspects of thehen’s oviduct after removal of preen gland, in youngand old age.

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This study was carried out on histomorphologicalstructure and micrometry of the oviduct before andafter extirpation of the preen gland in relation to eggproduction in chicken (Gallus gallus domesticus). Theexperimental 100 White Leghorn chickens (layerbirds/ hens) were from two age group i.e. (A) YoungHen aged 32 wks old and (B) Spent Hen aged 90 wksold, comprising 50 birds in each group. The younghens were again subdivided into Group I - Startcontrol group (10 birds), Group II - End control group(20 birds) and Group III - preen gland extirpatedgroup (20 birds). The spent hens were againsubdivided into Group IV - Start control group (10birds), Group V - End control group (20 birds) andGroup VI - preen gland extirpated group (20 birds).

They were reared in cage housing system ofCalifornia type with two tiers and four rows in threepens. The standard procedure of scientific feeding andmanagement were followed throughout theexperiment. Sixteen hours of light and eight hours ofdarkness was provided to the birds throughout theexperimental period of six weeks.

Surgical excision of the uropygeal gland wasperformed under local anesthesia and with the use ofelectro-cautery and coagulant to maintain hemostasisand minimize damage to the retrices. A fusiformincision was made at base of the gland and the glandwas bluntly dissected. After uropygealectomy all thebirds were given 15 days rest periods to recover stressand to returns in normal production.

Preen gland were surgically removed from 40birds of Group-III and Group-VI. Afteruropygealectomy all the birds were given 15 days restperiods to recover stress and to returns in normalproduction. Ten birds from each groups weresacrificed for histological study and from each partof the oviduct, 3 – 5 mm dimension tissue pieces werecut across the centre and preserved in 10 % neutralbuffered formalin for 24 hours at room temperature.These tissue pieces were then treated with routinemethod of dehydration in ascending grades of alcohol(ethanol), cleared in xylene / cedar wood oil andembedded in paraffin wax of the low melting point asper Drury et al. (1967). The paraffin blocks weresectioned at 3 to 5 micron thickness.

The micrometrical observations were carried outfor the various components of the each part of theoviduct in all group before and after extirpation ofthe preen gland as per method of Culling (1969). Thestained tissue sections of various segments of theoviduct were micro-photographed.


Histology of the various parts of the oviduct:

The oviduct in fowl represents the entirereproductive duct. The left oviduct was consisted ofinfundibulum, magnum, isthmus, uterus and vagina,while the right oviduct was rudimentary. Theseobservations were in accordance with the reportedfindings of Drapper et al. (1971), Eurell and Frappier(2006) and Khokhlov and Kuznetcov (2007). Ghuleet al. (2010b) in quails and Saber et al. (2010). reportedthat each of this division had two morphologicalessential elements for egg formation and movementof the egg along the oviduct i.e. glandular epitheliallining which secrete all the parts of egg outside theyolk-filled oocyte and muscular layer, which supportthe oviduct and protect the eggs. The wall of theoviduct was composed of four basic layers viz., tunicamucosa, tunica submucosa, tunica muscularis andtunica serosa from within outwards.

Infundibulum

Tunica mucosa of infundibulum was pale incolour. The mucosal and submucosal folds werethrown into low longitudinal ridges in the funnelportion which became progressively higher and higherin tubular portion from Group I to VI. The mucosa-submucosa was thrown into tall primary folds andsecondary folds (Group III and VI). This differencemight be due to progressive development of themucosal folds due to effect of the extirpation of preengland. The secondary folds were prominent inposterior end of tubular part. The anterior or funnel-shaped part was lined by simple columnar epithelium.The posterior tubular part or chalaziferous region waslined by pseudo-stratified columnar ciliatedepithelium (Fig.1) all over the folds except at thebottom of these folds where simple columnarepithelium was present and contained few goblet cells

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(Fig. 2). The cell height was low in Group I and IVwhere as it was more in preen gland extirpated GroupIII and VI. At the base of these mucosal folds, thesurface epithelium was reflected upwards to take ‘U’turn or in folded into new mucosal fold which formeda narrow depression called pits or glandular grooves.These grooves represent the sperm-host gland of theinfundibulum (Figs. 3) which were lined by the largesecretory cells without cilia with rounded basal nuclei.Dense secretory material was seen inside thecytoplasm bellow the apical or luminal border of thecells. These findings are in accordance with Surface(1912) who mentioned that these pits are the glandulargrooves. These observations are in concurrence withthe observation of Aitkin and Johnson (1963), Wyburnet al. (1970) and Eurell and Frappier (2006).

The funnel shaped part of the infundibulumshowed short mucosal folds without active secretorycells or glands, except sperm-host gland (Fig. 3) whichwere similar with the findings of Mohammadpour andKeshtmandi (2008) in turkey and pigeon. Theglandular grooves form the sperm-host gland of theinfundibulum, were lined by the secretory epithelium.The sperm were stored inside the lumen of theseglands matched the observation of Bakst (1978).

In the present study, four kinds of epithelial cellswere seen or distinguished in the epithelium as below:1) Columnar ciliated epithelium which were

nonsecreting in nature and located in the surfaceepithelium over the mucosal folds (Fig. 1).

2) Non-ciliated mucus secreting or goblet cells

spread between columnar ciliated epithelial cellsin the surface epithelium (Fig. 2).

3) Large secretory cells at the base of these mucosalfolds, which lined the lumen of sperm-host gland(Fig. 3) and

4) Tubular gland cells seen in the tubular gland (Fig.4).

Aitkin and Johnston (1963) reported that, ciliaof columnar cells were tubular in shape having lengthand width as 3-4 µm and 0.2 µm, respectively. Presentfindings are similar to the description of Hodges(1974) and Eurell and Frappier (2006).

The secretory and tubular gland cells were seenin active stage of secretion with numerous secretoryblebs pouring their secretion into the lumen of theinfundibulum. The secretory blebs are more prominentin Group III and VI (Fig. 2, 3 and 4). This differencemight be due to progressive increase in the secretoryactivity of the surface epithelium due to the extirpationof preen gland. These cells secreted chalaziferouslayer over the yolk.

The mucosal folds suddenly changed at thejunction of infundibulum and magnum in height andthickness. The height of mucosal folds was noticedlow in infundibulum which immediately becomes 3-4 times higher in magnum.

Tunica submucosa consisted of loosely arrangedconnective tissue with collagenous and reticular fibers,plasma cells and irregular shaped cells. The bloodvessel were fine and numerous. The submucosa were

Fig. 1 : Microphotograph of infundibulum from gr-IIshowing A – nucleus of ciliated cells, B – surface epithelium,C – secretary blebs and D – blood vessel. (HE, 400X).

A

B

D C A

Fig. 2 : Microphotograph of goblet cells in infundibulumfrom gr-V showing A – rounded basal nucleus of non-ciliated goblet cell, B – secretary epithelium, C – secretaryblebs and D – blood vessel. (HE, 400X).

A B

C D

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seems to be extended into primary and secondarymucosal folds. Below the surface epithelium, thetubular glands were seen increases in numbers whichwere extending from glandular grooves and groupedin a linear fashion which represent tubular gland ofthe magnum. The submucosa was found thickest inGroup III and thinnest in Group V. The gland cellsshowed large infranuclear granular endoplasmicreticular space filled with homogeneous material oflow electron density responsible for storage ofsynthesized protein (Group VI). This difference mightbe due to progressive increase in the secretory activityof the pyramidal cells of the tubular gland due to effectof the extirpation of preen gland. These findings aresimilar to the description of Hodges (1974), Getty(1975) and Eurell and Frappier (2006).

Tunica musculosa were consisted ofprogressively increasing longitudinal smooth musclefiber bundles towards the tubular end. These findingsare similar to the description of Hodges (1974), Getty(1975) and Eurell and Frappier (2006) who stated thatscattered muscle bundles of longitudinal smoothmuscles. The thickest tunica muscularis was noticedin Group III and VI while thinnest was noticed inGroup I and V. The muscle fiber bundles werearranged into a thin, inner circular and a thick, outerlongitudinal layer.

Tunica serosa of infundibulum comprised ofvarying amount of loose connective tissue, the bloodvessels and nerves. The tunica serosa consisted of

mesothelial lining covering the connective tissue layerwhich was divisible into two layers. The inner layerwas close to tunica muscularis which was relativelydenser and rich in elastic fibers than the outer layer.

Magnum : Magnum was longest portion of theoviduct. Its wall was thick, folded and lined bypseudostratified columnar ciliated epithelium (Fig. 7).

In the tunica mucosa of magnum, the mucousmembrane was projected into longitudinal, thick spiralfolds. It was thick in Group III and V and thin in GroupI. The number of mucosal folds was more in Group Iand IV and less in Group III and VI. The number ofmucosal folds decreased in Group III and VI butincreased in mucosal height and subdivided intosecondary and tertiary folds. This difference mightbe due to progressive development of the mucosalfolds and increase in the secretory activity of thesurface epithelium due to the effect of the extirpationof preen gland. The mucosa was deeply folded andeach fold was broad and bifid. The epithelium of themagnum consisted of columnar ciliated cells andgoblet cells found one after the other (Fig. 7). Thesecells were observed tall in Group I, II, IV and V, andtallest in Group III and VI. This difference might bedue to progressive development of the mucosal foldsand increase in the secretory activity of the surfaceepithelium due to the effect of the extirpation of preen

Fig. 3 : Microphotograph of sperm-host gland of theinfundibulum from gr-III showing A – basal nucleus, B –surface epithelium, C – secretary blebs and D – Lumen ofSperm host gland. (HE, 400X).

A

B

C D

Fig. 4 : Microphotograph of infundibulum from gr-VIshowing A – basal nucleus, B – secretary pyramidalepithelium of tubular glands, C – secretary blebs and D –lumen of tubular glands, E - blood vessel and F - fibroblast.(HE, 400X)

A

B

C

E

D

F

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gland. The nuclei of columnar ciliated cells wererounded and located in center of the cells. The nucleiof goblet cells were flattened dorso-ventrally andlocated near basal border of the cells. The cytoplasmcontained full of larger granules might containingmucoid substance, which indicated presence of acidmucopolysaccharides demonstrated by strongAldehyde Fuschin- alcian blue activity. This differencemight be due to progressive increase in the secretoryactivity of the surface epithelium due to effect of theextirpation of preen gland. The mucosa of theinfundibulum and magnum were separated by anarrow, white, transparent epithelial band which werelined by simple columnar epithelium and devoid ofthe glands. This observation is in accordance with thefindings of Surface (1912), Richardson (1935),Hodges (1974) and Eurell and Frappier (2006),Giersberg (1922), Bradley (1928), Ozen et al. (2009)

Tunica submucosa showed loose connectivetissue with abundance of collagen fibers, bloodcapillaries and was packed with long, branched, coiledtubular gland. Tunica submucosa of the magnum wasthickest in Group III and VI than Group I and IV. Thecells of the glands were pyramidal in shape andcontained course granules containing protein granules(Fig. 5 & 7). This difference might be due toprogressive increase in the tubular gland size due tothe effect of the extirpation of preen gland. Thebranched convoluted tubular glands were extendedinto the submucosa due to greatest development inthe magnum in Group III and VI. This difference mightbe due to progressive increase in the tubular glandsize due to effect of the extirpation of preen gland.The glandular cells contained large eosinophilicgranules with small flattened basal nuclei. The ductsopened anywhere on the luminal surface but weredifficult to see in ordinary histological sections butcould be observed after their secretion has beenreleased. The cells of these glands were packed withtheir secretion. The lumen of the gland and the inter-glandular connective tissue was invisible immediatelybefore ovulation. However, the lumen and the outlinesof the individual glands were more easilydistinguished after discharge of the secretions. In lastfew centimeters of the magnum the folds and thetubular glands were much reduced and the glandularcells contained relatively abundant mucous. Thetubular glands were opened on the surface of the

mucosal folds and their openings were remainedhidden in between the ciliated columnar epithelial cells(Fig. 7). The lumen of these magnal tubes was narrowin albumen secreting phase and were wider inregression phase. During the albumen secreting phase,the secretory granules formed blebs at luminal surfaceof pyramidal cells which narrowed the lumen. Inregression phase, there were no secretory granules incell cytoplasm, therefore the lumen was wide.

These observations are in agreement with thereported findings of King and Mclelland (1975),Wyburn et al. (1973), Eurell and Frappier (2006),Ghule et al. (2010a).

Tunica muscularis layer or tunica musculosaconsisted of smooth muscle fibers arranged as innercircular and outer longitudinal layers. The muscularlayers were thicker and bundles were more clearlyarranged in definitive layers. These observations arein concurrence with the observation of Surface (1912),Eurell and Frappier (2006) and Ghule et al. (2010b).Romanoff and Romanoff (1949). Tunica serosa wasconsisted of very thin layer of loose connective tissueand covered by flat mesothelial cells. It was thinnestin Group I and IV, and thickest in Group III and VI.The increased in thickness was observed due toincreased collagenous fibers and adipose tissue. Thisdifference might be due to progressive increase in theage of birds along with their body size and due to theeffect of the extirpation of preen gland.

Isthmus :

The isthmus was a short, narrow, constricted partof the oviduct lies between the magnum and uterus.Its wall was consisted of tunica mucosa, submucosa,musculosa and serosa, from inside outwards. Similarobservations were reported by Hodges (1974) in fowl,Mehta et al. (2005), Eurell and Frappier (2006) andKanchana et al. (2006) in Guinea fowl and Saber etal. (2010) in ostrich.

Tunica mucosa of the isthmus was lined bypseudo-stratified columnar ciliated cells (Fig.6) mixedwith goblet cells. The height of the epithelium wasincreased in preen gland extirpated Group III anddecreased in Group VI. Mucosal folds were leaf-likeand unbranched. This difference might be due toprogressive increase in the secretory activity of thesurface epithelium due to effect of the extirpation of

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preen gland. The arrangement of mucosal ridges inthe isthmus became almost longitudinal instead ofspiral and the layer of tubular glands within the ridgeswas similar in appearance to that of the magnumalthough it was not so well developed. The tubulargland cells were filled with secretory granules.Microvilli were prominent containing amorphousmaterial which were condensed into a membrane fiberin the lumen of a tubular gland and reached to theluminal surface of the oviduct responsible for theformation of the basic fiber of the shell membrane.

The lamina propria and tunica submucosa were filledwith distended branched tubular glands (Fig.6 & 8).The mucosal folds were longitudinally oriented, whitein colour and were less thick than magnal folds. Thefolds were found thick in Group II and VI. The numberof folds was less in Group II and more in Group VI.

Similar was the finding of Surface (1912).Bradley (1928), Richardson (1935), Draper et al.(1971), Hodges (1974), Bakst (1978), Eurell andFrappier (2006), Kanchana et al. (2006), and Ghuleet al. (2010b).

Fig. 5 : Microphotograph of magnum from gr-IV showingA – nucleus of the secretary epithelium, B – secretorysubstance in lumen and C – tubular glands. (HE, 400X).

A

B

C

Fig. 6 : Microphotograph of isthmus from gr-I showingA – nucleus of ciliated cells, B – secretary blebs of gobletcell, C –tubular gland cells, D – lumen and E – opening ofduct of tubular gland with released secretion. F – lumenof tubular gland. (HE, 400X)

A B C D

A

Fig. 7 : Microphotograph of magnum in albumen secretingphase from gr-V showing A – goblet cells loaded withmucin granules, B – Ciliated epithelium, C – pale stainingsecretary albumen in lumen and D – tubular glands full ofovalbumen. (HE, 400X)

Fig. 8 : Microphotograph of isthmus from gr-III showingA – nucleus, B – secretary epithelium, C – secretary blebsand D – lumen. (HE, 400X)

A

B

C

D E

F

C D

B

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Tunica submucosa was composed of looseconnective tissue with blood capillaries, nerves andtubular glands. The submucosa of the isthmus wasmainly occupied by the distended branched tubularglands and was thick in Group II and VI. The cells ofthe gland were pyramidal in shape with secretorygranules. These observations are in agreement withthe findings of King and Mclelland (1975), Kanchanaet al. (2006c) in Guinea fowl. Tunica muscularis ofisthmus was similar to the magnum but thicker inGroup III and VI. The tunica muscularis was madeup of inner circular and outer longitudinal smoothmuscle fibers. These findings are similar to thedescription of Eurell and Frappier (2006) Kanchanaet al. (2006) in Guinea fowl. Tunica serosa consistedof loose connective tissue and mesothelium. Theadipose cells were more in Group VI than Group III.

Uterus :

The uterus was the thickest and widest part ofthe oviduct, was grayish in colour and held in positionby ventral ligament.

Tunica mucosa of the uterus was lined by tallpseudo-stratified columnar ciliated epithelium (Fig.9). The height increased slowly from Group I to III inyoung hens and from Group IV to VI in spent hens.The number of mucosal fold was increased in Group

III and VI. This difference might be due to progressivedevelopment of the mucosal folds due to effect of theextirpation of preen gland. The mucosal folds werelong and branched and arranged longitudinally(Fig. 10). The height of the columnar epitheliumdecreases but the secretory granules in the columnarepithelial cells and the cilia on the apical cells of thecolumnar epithelium were preserved. The mucosaformed in deep straight crypts which was wellvascularized with supporting cores formed byconnective tissue. It was responsible for calcareousshell formation.

The lamina epithelialis was composed of ciliatedand secretory cells. The luminal surface of the uteruscomposed of tall longitudinal mucosal folds whichhave lamina epithelialis resembled with that of theprevious segments.

These observations are in agreement with theobservation of Timo (1969). Philip et al. (2005) andOzen et al. (2009). These findings are similar to thefindings of Saber et al. (2010), (Ghule et al., 2010a).

Tunica submucosa was thick and composed ofloose connective tissue with collagen fibers, arterioles,blood capillaries and contained long, branched, coiledtubular glands (Fig. 10). The cells of these glands werepyramidal in shape (Fig. 11) with scattered granulesand vacuole in the cytoplasm. Very fine collagenousand reticular fibers surrounded these tubular glands.

E

D

C

A

F

G H

I J

Fig. 9 : Microphotograph of uterine wall from gr.-VIshowing – A – pseudo-stratified ciliated epithelium, B –goblet cell, C – secretary blebs, D – BV, E – tubular glandepithelium. F – basement membrane. G – lymphocyte. H –fibroblast, I – collagen fibers and J – plasma cell. (HE,400X).

Fig. 10 : Microphotograph of developing tubular glands ofuterus from gr.-VI showing - A – mucosal fold 3060 µmin height, B – surface epithelium, C – uterine lumen,D – glandular epithelium and E – lamina propria. (HE,40X)

E

D C

B

A

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Under high power, the connective tissue showedplasma cells, fibroblast, lymphocytes, arteries andveins. The thickness was noticed more in Group IIand IV. These findings are similar to the descriptionof Ozen et al. (2009) and Ghule (2010b). During eggformation, these vacuoles regressed following theformation of extensive rough endoplasmic reticulumwhich was reverse also. The disintegrated materialsfound in the vacuoles were derived from the granules.

Tunica muscularis was made up of inner thickercircular and outer thinner longitudinal muscle fibersseparated by stratum vascularae - a loose connectivetissue layer which contained numerous blood vessels.The inner circular layer was thick at the junction withvagina and forms a sphincter. Tunica serosa was thinin Group I and IV which increased in thickness inGroup III and VI. It was thickest at the attachment ofinferior ligament of the oviduct. These observationsare in accordance with the reports of Hodges (1974),Getty (1975) and Eurell and Frappier (2006).

Vagina :

Vagina was the short, thick-walled part of theoviduct which was ‘S’ shaped.

Tunica mucosa was lined by pseudo-stratified

columnar ciliated epithelium and goblet cells. Thegoblet cells increased in numbers from Group I to VIwhile height of epithelial cells was more in Group IIIand V. The mucosal folds were longitudinally oriented,narrow, branched and irregular in shape. Thesemucosal folds were devoid of tubular glands. Theseincreased in height from Group I to Group VI. Thesperm-nest glands (Fig. 12) were numerous locatedin the mucous villi at the utero-vaginal junction, whichstore the sperms. These glands were lined by simplecolumnar epithelial cells. The cells of the tubule werenon-ciliated type with round or oval nucleus in theirbasal portion. The cytoplasm was finely or coarselyvacuolated due to lipid inclusions. Similar findingswere noted by Akira and Nakamura (2001), Thomas(2003), Kanchana et al. (2006), De Ocampo (2008),Ozen et al. (2009), and Ghule (2010b).

Tunica submucosa consisted of loose connectivetissue containing lymphocyte and granulocytes. It wasrecorded thickest in Group III and VI. The tunicamuscularis of the vagina showed inner thickcircularly oriented and outer longitudinal muscularlayer. The thickness of muscular layer was found morein Group III and VI than I and IV, respectively. Thisdifference might be due to the effect of the extirpationof preen gland. Tunica serosa showed diffused loose

Fig. 12 : Microphotograph of vagina from gr.-VI showing- A – mucosal folds about 3060 µm in height, B – spermhost gland and C – lumen of vagina. (HE, 40X)

B

A

C

F C

B

A

E

G

D

Fig. 11 : Microphotograph of tubular gland cells of uterusbefore egg shell formation from gr.-VI showing - A – basalnucleus of simple columnar cells or Type I cell, B – ciliaat apical border of secretary epithelium, C – lumen of thetubular gland, D – Secretory material from apical borderof Type II cell, E - artery, F – vein engorged with bloodand G - fibroblast. (HE, 400X).

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connective tissue cells and mesothelium. It wasthickest in all the parts of the oviduct. The higherthickness was recorded in Group III and VI. Similarobservations were reported by Hodges (1974), Getty(1975) and Eurell and Frappier (2006).

Micrometry of the various parts of the oviduct :

The micrometrical observations of differentparameters of oviduct of various groups of WLHlayers were recorded.

Micrometry of the infundibulum :

The statistical analyses of various parameters ofmicrometrical observations of infundibulum in variousgroups are given in Table No. 1.

Height of the epithelium : In young hens, theaverage height of the surface epithelium, at tubularportion of infundibulum, ranged from 16.84 ± 0.57,18.29 ± 0.97 and 20.98 ± 0.40 µm in Group I to III,respectively. In spent hens, the average height of thesurface or luminal epithelium, at tubular portion ofinfundibulum, ranged from 17.32 ± 1.28, 19.28 ± 0.57and 23.18 ± 0.98 µm in Group IV to VI, respectively.

The mean height of the epithelium in preen glandextirpated groups were recorded highest than theircontrols. This difference might be due to theprogressive increase in the secretory activity of thesurface epithelium and higher metabolic rate in preengland extirpated groups of spent hens and also theeffect of the extirpation of preen gland which relatedto increase in egg production. These observations arein agreement with the reported findings of Khokhlovand Kuznetcov (2007) and Khokhlov (2008). Themean differences in the height of the epithelium ininfundibulum of the oviduct from different groupswere statistically significant at 1 % level ofsignificance. Further, it can be concluded that,extirpation of the preen gland at age groups of 32 and90 weeks, increases the height of the epithelium.

Height of Villi : In young hens, the averageheight of the villi, at tubular portion of infundibulum,ranged from 911.70 ± 49.28, 926.10 ± 39.99 and1073.70 ± 46.79 µm in Group I to III, respectively. Inspent hens, the average height of the villi, at tubular

portion of infundibulum, ranged from 1276 ± 86.82,1326.60 ± 115.74 and 1659.60 ± 82.93 µm in GroupIV to VI, respectively.

The mean height of villi in preen gland extirpatedGroup III and VI recorded higher than start and endcontrol groups. This difference might be due toprogressive development of the mucosal folds and theeffect of the extirpation of preen gland. Khokhlov andKuznetcov (2007) noticed that, the height of themucosal folds/ villi at the age of 540 days in BLHwas 146.80 ± 16.20 µm. Khokhlov (2008) whoobserved that, the height of the mucosal folds whichwas 946.4 ± 51.3 ìm at the age of 210 days and146.8±16.2 ìm at the age of 540 days in hen.Mohammadpour and Keshtmandi (2008) in pigeonand turkey stated that, the length of the mucosal foldswas 500.47 ± 57.70 µm and 125.25 ± 28.80 µm,respectively.

The differences in the height of the villi ininfundibulum of the oviduct from different groupswere highly significant. Further, it can be concludedthat, extirpation of the preen gland at age groups of32 and 90 weeks, increases the height of the villi.

Number of villi : In young hens, the averagenumber of villi/mm2, at tubular portion ofinfundibulum, ranged from 4.08 ± 0.35, 4.24 ± 0.34and 4.08 ± 0.26 in Group I to III, respectively. In spenthens, the average number of villi/mm2, at tubularportion of infundibulum, ranged from 4.71 ± 0.33,5.02 ± 0.31 and 5.65 ± 0.48 in Group IV to VI,respectively.

The differences between the average values weresignificant at 5% level. The increase in number ofvilli might be due to increase in reproductive activityand also the extirpation of the preen gland therebyincreasing the surface area for secretion.

Number of Goblet cells : In young hens, theaverage number of goblet cells/mm2, at tubular portionof infundibulum, ranged from 57.31 ± 3.12, 61.23 ±2.82 and 75.83 ± 2.34 in Group I to III, respectively.In spent hens, the average number of goblet cells/mm2,at tubular portion of infundibulum, ranged from 66.41± 0.74, 73.63 ± 1.03 and 79.13 ± 0.94 in Group IV toVI, respectively.

The perusal of available review of literature onthe number of goblet cells in infundibulum was not

26


traceable for comparison. In the present study, thenumber of goblet cells in infundibulum wasprogressively increased with the advancement of ageand the highest mean number of goblet cells ininfundibulum was recorded in the Group III and VI.

The differences between the average values werehighly significant at 1 % level. This difference mightbe due to the effect of the extirpation of preen gland.The increased number of goblet cells might berequired to protect the mucosal surface of the oviductagainst mechanical irritation and propelling effectsof the increased muscular activity as reported bySturkie (1975).

This indicated that, extirpation of the preen glandat age groups of 32 and 90 weeks had favorable effectson number of goblet cells of the infundibulum.

Thickness of tunica mucosa : In young hens,the average thickness of tunica mucosa, at tubularportion of infundibulum, the range increased from1044.90 ± 69.55, 1027.80 ± 38.46 and 1126.80 ±48.69 µm in Group I to III, respectively. In spent hens,the average thickness of tunica mucosa, at tubularportion of infundibulum, the range increased from1300 ± 92.08, 1363.05 ± 115.68 and 1705.50 ± 83.28µm in Group IV to VI, respectively. The differencesbetween the average values were significant at 1%level. The thickness of mucosa was increased mainlybecause of increase in size and number of tubularglands and this difference might be due to the effectof the extirpation of preen gland.

Thickness of lamina propria : In young hens,the average thickness of lamina propria, at tubularportion of infundibulum, the range increased from19.80 ± 1.20, 79.20 ± 4.61 and 30.60 ± 1.47 µm inGroup I to III, respectively. In spent hens, the averagethickness of lamina propria, at tubular portion ofinfundibulum, the range increased from 33.30 ± 3.81,14.40 ± 1.99 and 23.40 ± 1.47 µm in Group IV to VI,respectively. The differences between the averagevalues were highly significant (p < 0.01).

Thickness of muscularis mucosae : In younghens, the average thickness of muscularis mucosae,at tubular portion of infundibulum, the range increasedfrom 21.60 ± 1.47, 36.60 ± 2.75 and 18 ± 1.90 µm inGroup I to III, respectively. In spent hens, the average

thickness of muscularis mucosae, at tubular portionof infundibulum, the range increased from 22.50 ±2.77, 23.40 ± 1.99 and 21.60 ± 1.47 µm in Group IVto VI, respectively. The differences between theaverage values were statistically significant at 1%level.

Thickness of tunica submucosa : In younghens, the average thickness of tunica submucosa, attubular portion of infundibulum, the range increasedfrom 53.10 ± 7.16, 32.40 ± 3.60 and 84.60 ± 4.07 µmin Group I to III, respectively. In spent hens, theaverage thickness of tunica submucosa, at tubularportion of infundibulum, the range increased from22.50 ± 1.50, 21.60 ± 1.47 and 26.10 ± 2.49 µm inGroup IV to VI, respectively. The differences betweenthe average values were significant at 1% level. Theincrease in thickness of tunica submucosa was due tocorresponding increase in the tubular glands and alsothe effect of the extirpation of preen gland.

Thickness of tunica muscularis : In younghens, the average thickness of tunica muscularis, attubular portion of infundibulum, the range increasedfrom 84.60 ± 7.37, 173.70 ± 4.85 and 199.80 ± 10.63µm in Group I to III, respectively. In spent hens, theaverage thickness of tunica muscularis, at tubularportion of infundibulum, the range increased from 225± 12.80, 207 ± 17.02 and 253.80 ± 30.26 µm in GroupIV to VI, respectively. The differences between theaverage values were highly significant (P<0.01). Agegroup comparison indicated a significant increase inthe thickness of tunica muscularis which was inaccordance with the increase in body size andphysiological activity related to more egg productiondue to the effect of the extirpation of preen gland.

Thickness of tunica serosa : In young hens, theaverage thickness of tunica serosa, at tubular portionof infundibulum, the range increased from 24.30 ±2.34, 173.70 ± 4.85 and 199.80 ± 10.63 µm in GroupI to III, respectively. In spent hens, the averagethickness of tunica serosa, at tubular portion ofinfundibulum, the range increased from 225 ± 12.80,207 ± 17.02 and 253.80 ± 30.26 µm in Group IV toVI, respectively. The differences between the averagevalues were significant at 1% level.

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Micrometry of the magnum :

The statistical analyses of various parameters ofmicrometrical observations of magnum in variousgroups are given in Table No. 2.

Height of the epithelium : In young hens, theaverage height of the surface or luminal epitheliumrange increased from 8.54 ± 0.41 µm, 21.96 ± 0.63µm and 25.62 ± 0.83 µm in Group I to III, respectively.In spent hens, the average height of the surface orluminal epithelium range increased from 18.79 ± 1.21µm, 18.54 ± 1.32 µm and 22.94 ± 1.10 µm in GroupIV to VI, respectively.

This observation is in accordance with thefindings of Richardson (1935), Wyburn et al. (1970),Sandoz (1971) and Sharma and Duda (1986),Khokhlov and Kuznetcov (2007).

The mean differences in the height of theepithelium in magnum of the oviduct from differentgroups were statistically significant at 1 % level ofsignificance. Further, it can be concluded that,extirpation of the preen gland at age groups of 32 and90 weeks, increases the height of the epithelium.

Height of Villi : In young hens, the averageheight of the villi of magnum ranged from 1154.70 ±78.05, 1305 ± 47.59 and 2135.70 ± 151.62 µm inGroup I to III, respectively. In spent hens, the averageheight of the villi, at tubular portion of magnum,ranged from 2618.10 ± 117.39, 2866.50 ± 172.30 and2853.90 ± 174.11 µm in Group IV to VI, respectively.The differences between the average values werehighly significant (p<0.01). These observations arein agreement with the reported findings of Romanoffand Romanoff (1949), Sharma and Duda (1986),Khokhlov and Kuznetcov (2007).

Number of villi : In young hens, the averagenumber of villi/mm2 of magnum ranged between 2.67± 0.24, 2.20 ± 0.26 and 2.04 ± 0.24 in Group-I to III,respectively. In spent hens, the average number ofvilli/mm2, ranged between 2.36 ± 0.26, 2.20 ± 0.26and 2.04 ± 0.24 in Group IV to VI, respectively. Thedifferences between the average values werestatistically non-significant. The decrease in numberof villi might be due to increase in secondary andtertiary mucosal folds due to the effect of the

extirpation of preen gland thereby increasing thesurface area for secretion. Khokhlov and Kuznetcov(2007) noticed that, the height of the mucosal folds/villi at the age of 540 days in BLH was 2057.10 ±50.60 µm.

Number of Goblet cells : In young hens, theaverage number of goblet cells/ mm2, ranged from44.75 ± 0.97, 52.91 ± 1.35 and 73.63 ± 1.99 in GroupI to III, respectively. In spent hens, the average numberof goblet cells/mm2, ranged from 58.72 ± 1.00, 63.43± 1.59 and 60.45 ± 1.85 in Group IV to VI,respectively. The differences between the averagevalues were statistically significant at 1% level. Theincreased number of goblet cells might be required toprotect the mucosal surface of the oviduct againstmechanical irritation and propelling effects of theincreased muscular activity as reported by Sturkie(1975) and Hodges (1974).

Thickness of tunica mucosa : In young hens,the average range of thickness of tunica mucosa inmagnum, was 1184.40 ± 80.28 µm, 1369.80 ± 44.90µm and 2157.30 ± 151.96 µm in Group I to III,respectively. In spent hens, the average range ofthickness of tunica mucosa was 2661.30 ± 11.64 µm,2925.90 ± 97.14 µm and 2902.50 ± 174.04 µm inGroup IV to VI, respectively. The differences betweenthe average values were highly significant. Thethickness of mucosa was increased mainly becauseof reduction in the numbers of villi and increasedsecondary and tertiary mucosal folds due to the effectof the extirpation of preen gland.

Thickness of lamina propria : In young hens,the average range of thickness of lamina propria inmagnum, ranged 12.60 ± 1.47, 30.60 ± 1.47 and 13.50± 1.50 µm in Group I to III, respectively. In spenthens, the average range of thickness of lamina propria,ranged from 17.10 ± 2.49, 21.60 ± 1.99 and 21.60 ±1.47 µm in Group IV to VI, respectively. Thedifferences between the average values weresignificant at 1% level.

Thickness of muscularis mucosae : In younghens, the average range of thickness of muscularismucosae, was 10.58 ± 1.20, 24.30 ± 2.70 and 11.70 ±1.37 µm in Group I to III, respectively. In spent hens,

28


the average range of thickness of muscularis mucosae,was 16.20 ± 1.80, 29.70 ± 1.37 and 22.50 ± 1.50 µmin Group IV to VI, respectively. The differencesbetween the average values were highly significant.

Thickness of tunica submucosa : In younghens, the average range of thickness of tunicasubmucosa, was 14.40 ± 1.47, 21.60 ± 1.47 and 26.10± 2.49 µm in Group I to III, respectively. In spenthens, the average range of thickness of tunicasubmucosa, increased from 18.90 ± 1.62, 24.30 ± 2.34and 32.40 ± 1.47 µm in Group IV to VI, respectively.The differences between the average values weresignificant at 1% level. The increase in thickness oftunica submucosa was the result of correspondingincrease in the tubular glands due to the effect of theextirpation of preen gland. These observations aresimilar with Surface (1912), Wyburn et al. (1970),Hodges(1974), King and Mclelland (1975), Gopinathand Hafeezuddin (1980), Sharma and Duda (1986)in Mallard and Thomas (2003).

Thickness of tunica muscularis : In younghens, the average range of thickness of tunicamuscularis was 34.20 ± 2.24, 70.10 ± 4.12 and 45 ±2.32 µm in Group I to III, respectively. In spent hens,the average range of thickness of tunica muscularis,increased from 133.20 ± 5.16, 128.70 ± 4.25 and117.90 ± 3.90 µm in Group IV to VI, respectively.The differences between the average values werehighly significant. Age group comparison indicated asignificant increase in the thickness of tunicamuscularis which was in accordance with the findingsof Sharma and Duda (1986).

Thickness of tunica serosa : In young hens, theaverage range of thickness of tunica serosa, was 16.20± 7.10, 24.30 ± 1.92 and 25.20 ± 2.24 µm in Group-Ito III, respectively. In spent hens, the average rangeof thickness of tunica serosa, was from 21.60 ± 1.74,22.50 ± 1.50 and 31.50 ± 1.50 µm in Group IV to VI,respectively. The differences between the averagevalues were significant at 1% level. Theseobservations are similar to those of Sharma and Duda(1986) reported in Mallard, the thickness of the serosawas 07.10 ± 0.02 µm.

Micrometry of the isthmus :

The statistical analyses of various parameters ofmicrometrical observations of isthmus in variousgroups are given in Table No. 3.

Height of the epithelium : In young hens, theaverage range of height of the luminal epithelium was17.81 ± 2.12, 34.40 ± 1.64 and 37.09 ± 0.61 µm inGroup I to III, respectively. In spent hens, the averagerange of height of the luminal epithelium was 33.67± 0.61, 29.77 ± 2.15 and 30.36 ± 1.16 µm in GroupIV to VI, respectively. The differences between theaverage values were highly significant. This differencemight be due to progressive increase in the secretoryactivity of the surface epithelium and the effect of theextirpation of preen gland. These observations are inaccordance with those by Wyburn et al. (1970),Sharma and Duda (1987), Khokhlov and Kuznetcov(2007).

Height of Villi : In young hens, the average rangeof height of the villi was 1048.50 ± 97.89, 1298 ±9.30 and 1213.20 ± 60.36 µm in Group I to III,respectively. In spent hens, the average range of heightof the villi was from 1544.40 ± 115.14, 1751.40 ±54.70 and 1799.10 ± 54.98 µm in Group IV to VI,respectively. The differences between the averagevalues were significant at 1% level.

Number of villi : In young hens, the averagerange of number of villi/mm2 of isthmus, was 5.34 ±0.26, 4.08 ± 0.26 and 5.18 ± 0.34 in Group I to III,respectively. In spent hens, the average range ofnumber of villi/mm2, was from 3.93 ± 0.26, 4.55 ±0.37 and 5.50 ± 0.26 in Group IV to VI, respectively.The differences between the average values werehighly significant. The increase in number of villimight be due to increase in reproductive activity withadvancement of age thereby increasing the surfacearea for secretion.

Number of Goblet cells : In young hens, theaverage range of number of goblet cells/mm2, was59.03 ± 2.25, 69.55 ± 1.39 and 82.27 ± 1.80 in GroupI to III, respectively. In spent hens, the average rangeof number of goblet cells/mm2, was from 47.89 ± 1.39,

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75.36 ± 5.15 and 79.29 ± 1.47 in Group IV to VI,respectively. The differences between the averagevalues were highly significant. The increased numberof goblet cells might be required to protect the mucosalsurface of the oviduct against mechanical irritationand propelling effects of the increased muscularactivity as reported by Sturkie (1975) and Hodges(1974).

Thickness of tunica mucosa : In young hens,the average range of thickness of tunica mucosa was1084.50 ± 97.19, 1349.10 ± 9.34 and 1254.60 ± 60.94µm in Group I to III, respectively. In spent hens, theaverage range of thickness of tunica mucosa was2661.30 ± 11.64, 2925.90 ± 97.14 and 2902.50 ±174.04 µm in Group IV to VI, respectively. Thedifferences between the average values weresignificant at 1% level. The thickness of mucosa wasincreased mainly because of increase in the numbersof villi.

Thickness of lamina propria : In young hens,the average range of thickness of lamina propria inisthmus, was 16.20 ± 2.24, 27 ± 1.90 and 19.80 ±1.20 µm in Group I to III, respectively. In spent hens,the average range of thickness of lamina propria, was21.60 ± 3.06, 21.60 ± 1.47 and 21.60 ± 1.47 µm inGroup IV to VI, respectively. The differences betweenthe average values were statistically significant.

Thickness of muscularis mucosae : In younghens, the average range of thickness of muscularismucosae, was 19.80 ± 1.80, 21.60 ± 1.99 and 20.70 ±1.37 µm in Group I to III, respectively. In spent hens,the average range of thickness of muscularis mucosae,was 19.80 ± 2.24, 24.30 ± 1.92 and 22.50 ± 1.50 µmin Group IV to VI, respectively. The differencesbetween the average values were statistically non-significant.

Thickness of tunica submucosa : In younghens, the average range of thickness of tunicasubmucosa, was 16.20 ± 2.94, 36 ± 1.90 and 21.60 ±1.47 µm in Group I to III, respectively. In spent hens,the average range of thickness of tunica submucosa,increased from 28.80 ± 2.24, 24.30 ± 1.92 and 36 ±2.23 µm in Group IV to VI, respectively. Thedifferences between the average values were highly

significant. The increase in thickness of tunicasubmucosa was due to corresponding increase in thetubular glands.

Thickness of tunica muscularis : In younghens, the average range of thickness of tunicamuscularis, was 173.70 ± 10.74, 256.50 ± 8.72 and656 ± 25.28 µm in Group I to III, respectively. Inspent hens, the average range of thickness of tunicamuscularis, was 352.80 ± 35.44, 336.60 ± 13.02 and478.80 ± 17.43 µm in Group IV to VI, respectively.The differences between the average values werestatistically significant at 1% level.

Thickness of tunica serosa : In young hens, theaverage range of thickness of tunica serosa, was from21.60 ± 1.47, 32.40 ± 4.07 and 29.70 ± 1.92 µm inGroup I to III, respectively. In spent hens, the averagerange of thickness of tunica serosa, was 71.10 ± 3.41,35.10 ± 3.41 and 45 ± 5.02 µm in Group IV to VI,respectively. The differences between the averagevalues were highly significant.

Micrometry of the uterus :

The statistical analyses of micrometricalparameters of micrometrical observations of uterusin various groups are given in Table No. 4.

Height of the epithelium : In young hens, theaverage range of height of the luminal epithelium wasfrom 8.78 ± 0.54, 18.30 ± 0.41 and 27.33 ± 0.61 µmin Group I to III, respectively. In spent hens, theaverage range of height of the luminal epithelium was17.08 ± 1.26, 25.38 ± 1.27 and 25.86 ± 1.55 µm inGroup IV to VI, respectively. The differences betweenthe average values were highly significant. Thisdifference might be due to progressive increase in thesecretory activity of the surface epithelium and alsothe effect of the extirpation of preen gland. Theseobservations are in agreement with the reportedfindings of Khokhlov and Kuznetcov (2007),Kanchana et al. (2006).

Height of Villi : In young hens, the average rangeof height of the villi, was 1754 ± 41.90, 1263 ± 31.10and 1330.20 ± 140.20 µm in Group I to III,respectively. In spent hens, the average range of height

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of the villi, was 2009.70 ± 185.29, 2039.40 ± 115.12and 1944.90 ± 50.12 µm in Group IV to VI,respectively. The differences between the averagevalues were significant at 1% level. Theseobservations are in agreement with the reportedfindings of Khokhlov and Kuznetcov (2007).

Number of villi : In young hens, the averagerange of number of villi/mm2, was 7.07 ± 0.26, 2.83± 0.21 and 5.34 ± 0.26 in Group-I to III, respectively.In spent hens, the average range of number of villi/mm2, was 3.14 ± 0.47, 4.24 ± 0.34 and 4.87 ± 0.37 inGroup-IV to VI, respectively. The differences betweenthe average values were highly significant. Theincrease in number of villi might be result of increasein reproductive activity might be due to effect of theextirpation of preen gland thereby increasing thesurface area for secretion.

Number of Goblet cells : In young hens, theaverage range of number of goblet cells/mm2, was47.57 ± 2.36, 53.85 ± 1.78 and 63.27 ± 1.72 in GroupI to III, respectively. In spent hens, the average rangeof number of goblet cells/mm2, was 58.88 ± 0.85,62.80 ± 0.74 and 66.10 ± 2.96 in Group IV to VI,respectively. The differences between the averagevalues were statistically significant at 1% level. Theincreased number of goblet cells might be required toprotect the mucosal surface of the oviduct againstmechanical irritation and propelling effects of theincreased muscular activity as reported by Sturkie(1975) and Hodges (1974).

Thickness of tunica mucosa : In young hens,the average range of thickness of tunica mucosa was1847.70 ± 77.65, 1313.10 ± 30.96 and 1386.90 ±139.31 µm in Group I to III, respectively. In spenthens, the average range of thickness of tunica mucosaincreased from 2052.90 ± 184.92, 2090.70 ± 115.33and 2001.60 ± 50.36 µm in Group IV to VI,respectively. The differences between the averagevalues were highly significant. The thickness ofmucosa was increased mainly because of reductionin the numbers of villi due to the effect of theextirpation of preen gland.

Thickness of lamina propria : In young hens,the average range of thickness of lamina propria, was

21.60 ± 1.47, 23.40 ± 1.99 and 32.40 ± 2.75 µm inGroup I to III, respectively. In spent hens, the averagerange of thickness of lamina propria, was 20.70 ± 1.37,27.00 ± 2.68 and 34.20 ± 2.62 µm in Group IV to VI,respectively. The differences between the averagevalues were highly significant.

Thickness of muscularis mucosae : In younghens, the average range of thickness of muscularismucosae, was 30.60 ± 2.40, 24.30 ± 1.92 and 24.30 ±1.92 µm in Group I to III, respectively. In spent hens,the average range of thickness of muscularis mucosae,was 22.50 ± 1.50, 24.30 ± 2.34 and 24.40 ± 1.99 µmin Group IV to VI, respectively. The differencesbetween the average values were statistically non-significant.

Thickness of tunica submucosa : In younghens, the average range of thickness of tunicasubmucosa, was 27.00 ± 2.32, 33.30 ± 2.70 and 278.80± 2.24 µm in Group I to III, respectively. In spenthens, the average range of thickness of tunicasubmucosa, was 31.50 ± 2.77, 22.50 ± 1.50 and 26.10± 2.10 µm in Group IV to VI, respectively. Thedifferences between the average values werestatistically significant. The increase in thickness oftunica submucosa was due to corresponding increasein the tubular glands.

Thickness of tunica muscularis : In younghens, the average range of thickness of tunicamuscularis, was 1591.20 ± 13.47, 1386.90 ± 103.86and 1286.10 ± 117.85 µm in Group I to III,respectively. In spent hens, the average range ofthickness of tunica muscularis, was 1094.40 ± 139.01,1390.50 ± 30.98 and 1387.80 ± 17.43 µm in GroupIV to VI, respectively. The differences between theaverage values were significant at 1% level.

Thickness of tunica serosa : In young hens, theaverage range of thickness of tunica serosa, was 36.00± 2.68, 42.30 ± 4.04 and 54.00 ± 2.32 µm in Group Ito III, respectively. In spent hens, the average rangeof thickness of tunica serosa, was 22.50 ± 1.50, 62.10± 7.99 and 95.90 ± 5.23 µm in Group IV to VI,respectively. The differences between the averagevalues were highly significant.

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Micrometry of the vagina:

The statistical analyses of micrometricalparameters of micrometrical observations of vaginain various groups are given in Table No. 5.

Height of the epithelium : In young hens, theaverage range of height of the luminal epithelium was11.22 ± 0.40, 16.10 ± 0.54 and 19.79 ± 0.73 µm inGroup I to III, respectively. In spent hens, the averagerange of height of the luminal epithelium was 23.42± 0.98, 22.94 ± 1.46 and 25.86 ± 0.98 µm in GroupIV to VI, respectively. The differences between theaverage values were highly significant.

Height of Villi : In young hens, the average rangeof height of the villi in vagina, was 711.90 ± 53.54,1949.40 ± 168.31 and 2808 ± 133.93 µm in Group Ito III, respectively. In spent hens, the average rangeof height of the villi, was 2084.40 ± 177.00, 1976.40± 120.08 and 2415.60 ± 193.89 µm in Group IV toVI, respectively. The differences between the averagevalues were significant at 1% level.

Number of villi : In young hens, the averagerange of number of villi/mm2, was 7.85 ± 0.33, 6.12± 0.37 and 7.54 ± 0.90 in Group I to III, respectively.In spent hens, the average range of number of villi/mm2, was 5.65 ± 0.42, 9.42 ± 0.41 and 11.30 ± 0.39in Group IV to VI, respectively. The differencesbetween the average values were highly significant.The increase in number of villi might be due toincrease in reproductive activity with advancementof age thereby increasing the surface area forsecretion.

Number of Goblet cells : In young hens, theaverage range of number of goblet cells/mm2, was54.64 ± 1.48, 59.66 ± 3.23 and 67.04 ± 1.10 in GroupI to III, respectively. In spent hens, the average rangeof number of goblet cells/mm2, was 60.45 ± 2.26,61.70 ± 1.24 and 77.40 ± 1.19 in Group IV to VI,respectively. The differences between the averagevalues were significant at 1% level. The increasednumber of goblet cells might be required to protectthe mucosal surface of the oviduct against mechanicalirritation and propelling effects of the increasedmuscular activity as reported by Sturkie (1975) andHodges (1974).

Thickness of tunica mucosa : In young hens,the average range of thickness of tunica mucosa invagina, was 762.30 ± 52.58, 2093.40 ± 177.24 and2260.80 ± 52.90 µm in Group I to III, respectively. Inspent hens, the average range of thickness of tunicamucosa was 2223.90 ± 120.71, 2160.00 ± 121.06 and2508.30 ± 188.83 µm in Group IV to VI, respectively.The differences between the average values werehighly significant. This difference might be due toprogressive development of the mucosal folds due toeffect of the extirpation of preen gland.

Thickness of lamina propria : In young hens,the average range of thickness of lamina propria, wasfrom 18.00 ± 1.90, 25.20 ± 2.24 and 39.60 ± 1.47 µmin Group I to III, respectively. In spent hens, theaverage range of thickness of lamina propria, was21.60 ± 1.47, 33.30 ± 2.70 and 25.20 ± 2.42 µm inGroup IV to VI, respectively. The differences betweenthe average values were statistically significant at 1%level.

Thickness of muscularis mucosae : In younghens, the average range of thickness of muscularismucosae, was 34.20 ± 1.80, 25.20 ± 2.24 and 27.90 ±2.49 µm in Group I to III, respectively. In spent hens,the average range of thickness of muscularis mucosae,was 63.00 ± 8.80, 80.10 ± 3.66 and 96.30 ± 8.17 µmin Group IV to VI, respectively. The differencesbetween the average values were highly significant.

Thickness of tunica submucosa : In younghens, the average range of thickness of tunicasubmucosa, was 22.50 ± 1.50, 23.40 ± 1.47 and 27.90± 2.10 µm in Group I to III, respectively. In spenthens, the average range of thickness of tunicasubmucosa was 23.40 ± 1.99, 33.30 ± 2.34 and 36.90± 2.10 µm in Group IV to VI, respectively. Thedifferences between the average values weresignificant at 1% level. The increase in thickness oftunica submucosa was result of correspondingincrease in the tubular glands might be due to effectof the extirpation of preen gland.

Thickness of tunica muscularis : In younghens, the average range of thickness of tunicamuscularis, was 788.40 ± 31.36, 1358.00 ± 161.58and 1431.00 ± 211.64 µm in Group I to III,

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respectively. In spent hens, the average range ofthickness of tunica muscularis was 1242.00 ± 122.23,1527.30 ± 212.70m and 1670.40 ± 173.12 µm inGroup IV to VI, respectively. The differences betweenthe average values were highly significant.

Thickness of tunica serosa : In young hens, theaverage range of thickness of tunica serosa, was 77.40± 5.56, 116.10 ± 7.64 and 149.40 ± 5.88 µm in GroupI to III, respectively. In spent hens, the average rangeof thickness of tunica serosa, was 116.10 ± 7.03,356.40 ± 54.72 and 371.70 ± 17.29 µm in Group IVto VI, respectively. The differences between theaverage values were significant at 1% level.

CONCLUSIONS

The mucus membrane of infundibulum was palein colour and showed mucosal-submucosal folds,which were arranged longitudinally and becameprogressively higher towards tubular part of theoviduct. These folds were angular varied in size andheight in all the groups. The maximum average heightof the folds were highest in preen gland extirpatedGroup III and VI. The numbers of fold were increasedfrom Group I to III and from IV to VI. These primarymucosal folds were branched into secondary andtertiary folds, which increased the functional surfacearea of the oviduct.

The surface epithelium of all parts of oviductwas pseudostratified columnar ciliated in all thegroups, which became simple columnar ciliated cellalternate to goblet cells at the base of the folds. Theheight of the surface epithelium, number of gobletcells and thickness of tunica mucosa increasedsignificantly in preen gland extirpated group III andVI than their control Groups I, II, IV and V. Theluminal surface of the columnar cells showedmicrovilli. In young and spent hens, the thickness oflamina propria and muscularis mucosae, increasedfrom Group I to III and from Group IV to VI,respectively. It was highest in Groups III and VI. Thesperm host / nest glands were seen in all the groups.

The tunica submucosa of infundibulum, magnumand uterus increased in thickness and was thickest inpreen gland extirpated Groups III and VI. The collagenand reticular fibers in all the groups were coarse inthe submucosa than the other layers of the oviduct.

Elastic fibers were poorly developed in all the groups.Tunica muscularis consisted of an inner circular andouter longitudinal layer of smooth muscle fibersseparated by stratum vascularae in all regions of theuterus in all groups. The muscular layer was initiallythin which increased in thickness from Group I to IIIin young hens and from Group IV to VI in spent hens.The tunica serosa was thin in Group I and IV, whichincreased and found significantly thicker in GroupIII and VI.

The mucus membrane of magnum was brown incolour. The mucosal-submucosal fold orientedspirally, which suddenly increased in height and werebranched into secondary folds.

The glandular epithelium almost pyramidal inshape and their nuclei were located in the basal partof the cells. In the tubular glands, the goblet cells wereabsent. The lumen was narrow in albumen secretingphase and were wider in regression phase. Due topassing of albumen secreting phase, the secretorygranules formed blebs at luminal surface of pyramidalcells which narrowed the lumen. In regression phase,there were no secretory granules in cell cytoplasm,therefore the lumen was wide. The tunica musculariswas increased in thickness from Group I to III in younghens and from Group IV to VI in spent hens. Thetunica serosa was thin in Group I and IV, whichincreased and was significantly thicker in Group IIIand VI.

The mucus membrane of isthmus was lightbrown and showed circular band at the magnum-isthmus junction with out mucosal folds which weretubular in all the groups. The average height of thefolds were highest in preen gland extirpated GroupIII and VI. The numbers of folds were increased fromGroup I to III and from IV to VI. These primarymucosal folds gave rise secondary and tertiary folds.

The tunica submucosa varied in thickness indifferent part of the isthmus. The collagen and reticularfibers in all the groups were coarse in the submucosathan the other layers of the oviduct. Elastic fibers weredeveloped in all the groups. The muscular layer wasthick in Group III and VI. The tunica serosa wassignificantly thicker in Group II and VI.

The mucus membrane of uterus was dark brownin colour in all the groups. The mucosal-submucosalfolds were oriented longitudinally and were leaf-shaped. These folds varied in size and height which

33


showed Iry and IIry folds in all groups.The tunica muscularis increased in thickness in

different part of the uterus. The tunica serosa was thinin Group I and IV, which increased and wassignificantly thicker in preen gland extirpated GroupIII and VI.

All the micrometrical observations of the variousparameters of different parts of the oviduct increasedwith age and extirpation of the preen gland.

From the present results following conclusionswere drawn:● The mucosal folds were oriented longitudinally

throughout the oviduct except in magnum, whereit was spirally oriented.

● These folds were angular in infundibulum andleaf-shaped in uterus. The surface epithelium waspseudostratified in all the groups, which becamesimple columnar at the base of the folds.

● The sperm host glands were seen in all the groupsin infundibulum and vagina.

● In magnum, the tubular gland epithelium waspyramidal in shape and their nuclei were locatedat the basal part of the cells. Goblet cells wereabsent in the tubular glands.

● All micrometrical observations of the oviductincreased with age due to the extirpation of preengland.

REFERENCES

Aitken, R. N. C. and Johnston H. S. (1963). In “Thehistology of the fowl” Academic Press Inc. Ltd.24-28 Oval Road, London NW 1 Pp.: 354 – 356.

Akira, C. and Nakamura M. (2001). Acta Zoologica,82(4): 299 – 306.

Bakst, M. R. (1978). Poultry Sci. 57 (4): 1065 – 1069.

Bradley, O. C. (1928). J. Anat. 62(3): 339 – 345.

Culling, C. F. A. (1969). In “Handbook ofHistopathological technique (including museumtechnique).” 2nd Edn. Butter worth and Co.,Philadelphia and London Pp: 228 – 238 and 477 –479.

De Ocampo, G.D. (2008). Gross and microscopicanatomy of the reproductive organs of the female

Philippine native chicken (Gallus Gallus domesticus).Animal physiology – Reproduction.

Draper M. H., Davidson, M. F., Wyburn G. M. andJohnston H. S. (1971). The fine structure of the fibrousmembrane forming region of the isthmus of theoviduct of Gallus domesticus. Agricultural ResearchCouncil’s Poultry Research Centre, King’s Buildings,West Mains Road, Edinburgh EH9 3JS, and theDepartment of Anatomy, University of Glasgow,Glasgow, W. 2 Received November 24, 1971.

Drury, R. A. B., Wallington E. A. and Cameron, R.(1967). In “Carleton’s Histological technique” OxfordUniversity Press, London 5th Edn. Pp. 23 – 98.

Eurell, J. A. and Frappier B. L. (2006). In “ Dellmann’sTextbook of Veterinary Histology.” 7th edn. Williamsand Wilkins. Pp : 275 – 77.

Getty, R. (1975). In Sisson and Grossman’s “Theanatomy of the domestic Animals.” Vol. 2 WBSaunders Company, Philadelphia Pp.: 1945 – 1958.

Giraudeau, M., Czirják, G. A., Duval, C., Bretagnolle,V. and Eraud, C. (2010). Effect of RestrictedPreen-Gland Access on Maternal Self Maintenance andReproductive Investment in Mallards.

Giersberg, H. (1922). In “The histology of the fowl”Academic Press INC Ltd., Oval Road, London. Nw1,361: 24 – 28.

Ghule P. M., Gaikwad, S. G., Dhande, P. L., Lambate,S. B., Tiwari, S.S. and Ayana, R. (2010a). J. BombayVet. College 18 (1): 44 – 47.

Ghule P. M., Gaikwad, S.G., Dhande, P. L., Lambate,S. B., Patil, A.D., Tiwari, S. S. and Ayana, R. (2010b).Indian J. Vet. Anat. 22 (1): 40–42

Gopinath, S. and Hafeezuddin, M. (1980). J.Maharashtra Agri. Universities. 5 (2): 153 – 156.

Hodges, R. D. (1974). A Textbook of “The Histologyof the Fowl”. Academic press Inc., London: pp. 347 -386

Ismail, A. R. A. (2005). Effect of Iraqi method(uropyialactomy) on some physiological andreproductive traits of broiler breeder males. A

34


Ran

geM

ean

± SE

Ran

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ean

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Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

1H

eight

of t

he

epith

elium

(µm

)14

.64

- 19

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16.8

4 ±

0.57

d17

.08

- 21.

9618

.29

± 0.

97cd

19.5

2 - 2

1.96

20.9

8 ±

0.40

ab12

.20

- 21.

9617

.32

± 1.

28cd

17.0

8 - 2

1.96

19.2

8 ±

0.57

bc19

.52

- 29.

2823

.18

± 0.

98a

12.1

19.

87**

2H

eight

of V

illi (µ

m)

630

- 111

691

1.70

±

49.2

8c71

1.00

- 10

8092

6.10

±

39.9

9c67

5 - 1

188

1073

.70

± 46

.79c

765

- 175

512

96 ±

86

.82b

792

- 171

013

26.6

0 ±

115.

74b

1233

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7016

59.6

0 ±

82.9

3a18

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17.4

3 **

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umbe

r of v

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mm

23.

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6.28

4.08

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0.35

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±

0.34

3.14

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0.

26b

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284.

71 ±

0.

33ab

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855.

65 ±

0.

48a

24.5

23.

03*

4N

umbe

r of G

oblet

ce

lls /

mm

248

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57.3

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c47

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5061

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± 2.

82bc

67.5

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2.63

75.8

3 ±

2.34

a62

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- 70.

6566

.41

± 0.

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69.0

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7879

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± 0.

94a

8.95

18.4

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f tun

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muc

osa

(µm

)81

0 - 1

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1044

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27.8

0 ±

38.4

6d72

0 - 1

269

1126

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± 48

.69cd

783

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819

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713

63.0

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1278

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8a19

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11.2

1 **

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m)

18 -

2719

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63 -1

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61a

27 -

3630

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47bc

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33.3

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2723

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47cd

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18 -

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75a

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9.00

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22.5

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23.4

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3.9 *

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5432

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60c

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84.6

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22.5

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36.4

5 **

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m)

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117

84.6

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b16

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8625

3.80

±

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13.8

7 **

10Th

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ss o

f tu

nica

sero

sa (µ

m)

9.00

- 36

24.3

0 ±

2.34

a18

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30.6

0 ±

3.06

c36

- 90

63 ±

4.8

4c27

- 45

36.9

0 ±

2.10

c27

- 25

210

9.80

±

25.6

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614

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21.2

2 **

Tabl

e N

o. 1

: Sta

tistic

al a

naly

sis

of m

icro

met

rical

obs

erva

tions

of i

nfun

dibu

lum

of t

he o

vidu

ct o

f Whi

te L

egho

rn la

yers

(Gal

lus g

allu

s dom

estic

us)

Sr.

No.

Para

met

er

You

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ens

(32

wee

ks)

Spen

t hen

s (9

0 w

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)%

of

CV

F' V

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Star

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d co

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glan

d ex

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35


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Ran

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Ran

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Ran

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ean

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Ran

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ean

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Ran

geM

ean

± SE

1H

eight

of t

he

epith

elium

(µm

)7.

32 -

9.76

8.54

±

0.41

d19

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- 24.

4021

.96

± 0.

63b

21.9

6 - 2

9.28

25.6

2 ±

0.83

a9.

76 -

24.4

018

.79

± 1.

21c

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18.5

4 ±

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c17

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2822

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± 1.

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15.6

438

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2H

eight

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m)

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011

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78.0

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47.5

9c14

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2835

2135

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± 15

1.62

b18

90 -

2925

2618

.10

± 11

7.39

a16

20 -

3510

2866

.50

± 17

2.30

a16

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2853

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4.11

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7 **

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142.

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0.

261.

57 -

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2.04

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142.

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0.2

61.

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1.57

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433

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4N

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ce

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mm

239

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44.7

5 ±

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6652

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35d

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73.6

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3758

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54.9

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513

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2943

2661

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2574

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4 **

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153

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108

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108

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128.

70 ±

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135

117.

90 ±

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178.

87**

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24.3

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b18

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25.2

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21.6

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b18

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Tabl

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tistic

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naly

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met

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m o

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Leg

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ticus

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36


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Ran

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Ran

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Ran

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Ran

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817

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12c

24.4

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1.48

34.4

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31.7

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6.60

29.7

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6.60

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377

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51.4

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217

99.1

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7879

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1254

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1143

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8815

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3.87

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S

8Th

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21.6

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18 -

5432

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18 -

3629

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92bc

18 -

5471

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41bc

27 -

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ens

(32

wee

ks)

Para

met

er

Tabl

e N

o. 3

: Sta

tistic

al a

naly

sis

of m

icro

met

rical

obs

erva

tions

of i

sthm

us o

f the

ovi

duct

of W

hite

Leg

horn

laye

rs (G

allu

s gal

lus d

omes

ticus

)

Sr.

No.

Pree

n gl

and

extir

pate

d gr

oup-

VI

Star

t con

trol

gro

up

or G

roup

-IEn

d co

ntro

l gro

up o

r G

roup

-IIPr

een

glan

d ex

tirpa

ted

grou

p-III

St

art c

ontr

ol g

roup

or

Gro

up-IV

Spen

t hen

s (9

0 w

eeks

)

% o

f C

V

End

cont

rol g

roup

or

Gro

up-V

F' V

alue

NS

: Non

sign

ifica

nt,

*

: (P

d”

0.05

) an

d *

* : (

P d”

0.0

1)N

ote

: Mea

n w

ith a

t lea

st o

ne c

omm

on su

pers

crip

t do

not d

iffer

sign

ifica

ntly

37


Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

1H

eight

of t

he

epith

elium

(µm

)7.

32 -

12.2

08.

78 ±

0.

54c

17.0

8 - 1

9.52

18.3

0 ±

0.41

b24

.40

- 29.

2827

.33

± 0.

61a

9.76

- 21

.96

17.0

8 ±

1.26

b19

.52

- 31.

7225

.38

± 1.

27a

19.5

2 - 3

4.16

25.8

6 ±

1.55

a15

.54

50.1

2 **

2H

eight

of V

illi (µ

m)

1584

- 19

8017

54 ±

41

.90a

1170

- 14

0412

63.6

0 ±

31.1

0b40

5 - 1

755

1330

.20

± 14

0.20

b12

33 -

3060

2009

.70

± 18

5.29

a14

40 -

2664

2039

.40

± 11

5.12

a16

83 -

2160

1944

.90

± 50

.12a

19.4

210

.67 *

*

3N

umbe

r of v

illi /

mm

26.

28 -

7.85

7.07

±

0.26

a1.

57 -

4.71

2.83

±

0.21

d4.

71 -

6.28

5.34

±

0.26

b1.

57 -

6.28

3.14

±

0.47

d3.

14 -

6.28

4.24

±

0.34

c3.

14 -

6.28

4.87

±

0.37

bc23

.66

20.6

2 **

4N

umbe

r of G

oblet

ce

lls /

mm

231

.40

- 56

.52

47.5

7 ±

2.36

b47

.10

- 62.

8053

.85

± 1.

78b

54.9

5 - 7

0.65

63.2

7 ±

1.72

a54

.95

- 62.

8058

.88

± 0.

85a

59.6

6 - 6

5.94

62.8

0 ±

0.74

a58

.09

- 81.

6466

.10

± 2.

96a

16.3

98.

35**

5Th

ickne

ss o

f tun

ica

muc

osa

(µm

)16

38 -

2475

1847

.70

± 77

.65a

1134

- 14

4913

13.1

0 ±

30.9

6b46

8 - 1

791

1386

.90

± 13

9.31

b12

69 -

3096

2052

.90

± 18

4.92

a14

76 -

2718

2090

.70

± 11

5.33

a17

37 -

2214

2001

.60

± 50

.36a

19.5

59.

84**

6Th

ickne

ss o

f lam

ina

prop

ria (µ

m)

18 -

2721

.60

± 1.

47cd

18 -

3623

.40

± 1.

99cd

18 -

4532

.40

± 2.

75ab

18 -

2720

.70

± 1.

37d

9.00

- 36

27 ±

2.6

8bc18

- 45

34.2

0 ±

2.62

a23

.28

8.46

**

7Th

ickne

ss o

f m

uscu

laris

muc

osae

(

)

18 -

4530

.60

± 2.

4018

- 36

24.3

0 ±

1.92

18 -

3624

.30

± 1.

9218

- 27

22.5

0 ±

1.50

18 -

3624

.30

± 2.

3418

- 36

24.4

0 ±

1.99

25.9

61.

99N

S

8Th

ickne

ss o

f tun

ica

subm

ucos

a (µ

m)

18 -

3627

±

2.32

abc

18 -

4533

.30

± 2.

70a

18 -

3628

.80

± 2.

24ab

c18

- 45

31.5

0 ±

2.77

ab18

- 27

22.5

0 ±

1.50

c18

- 36

26.1

0 ±

2.10

bc25

.76

2.86

*

9Th

ickne

ss o

f tun

ica

mus

cular

is (µ

m)

1584

- 16

6515

91.2

0 ±

13.4

7a63

0 - 1

620

1386

.90

± 10

3.86

ab36

0 - 1

710

1286

.10

± 11

7.85

bc36

0 - 1

503

1094

.40

± 13

9.01

c11

61 -

1503

1390

.50

± 30

.98ab

1134

- 15

3013

87.8

0 ±

44.4

9ab19

.04

3.94

**

10Th

ickne

ss o

f tu

nica

sero

sa (µ

m)

18 -

4536

± 2

.68d

27 -

6342

.30

± 4.

04cd

45 -

6354

± 2

.32bc

18 -

27

22.5

0 ±

1.50

e36

- 12

662

.10

± 7.

99b

72 -

126

95.4

0 ±

5.23

a27

.35

31.6

8 **

End

cont

rol g

roup

or

Gro

up-II

Star

t con

trol

gro

up o

r G

roup

-IV

End

cont

rol g

roup

or

Gro

up-V

Pree

n gl

and

extir

pate

d gr

oup-

VI

Tabl

e N

o. 4

: Sta

tistic

al a

naly

sis

of m

icro

met

rical

obs

erva

tions

of u

teru

s of

the

ovid

uct

of W

hite

Leg

horn

laye

rs (G

allu

s gal

lus d

omes

ticus

)

Sr.

No.

F' V

alue

Star

t con

trol

gro

up

or G

roup

-IPa

ram

eter

You

ng h

ens

(32

wee

ks)

% o

f C

V

Spen

t hen

s (9

0 w

eeks

)Pr

een

glan

d ex

tirpa

ted

grou

p-III

NS

: Non

sign

ifica

nt,

*

: (P

d”

0.05

) an

d *

* : (

P d”

0.0

1)N

ote

: Mea

n w

ith a

t lea

st o

ne c

omm

on su

pers

crip

t do

not d

iffer

sign

ifica

ntly

38


Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

Ran

geM

ean

± SE

1H

eight

of t

he

epith

elium

(µm

)9.

76 -

12.2

011

.22

± 0.

40e

14.6

4 - 1

9.52

16.1

0 ±

0.54

d17

.08

- 24.

4018

.79

± 0.

73c

19.5

2 - 2

9.28

23.4

2 ±

0.98

ab19

.52

- 34.

1622

.94

± 1.

46b

21.9

6 - 3

1.72

25.8

6 ±

0.98

a9.

6467

.11 *

*

2H

eight

of V

illi (µ

m)

396

- 891

711.

90 ±

53

.54d

1242

- 28

8019

49.4

0 ±

168.

31c

1692

- 30

6028

08 ±

13

3.93

a10

44 -

2610

2084

.40

± 17

7bc13

95 -

2430

1976

.40

± 12

0.08

c14

40 -

2988

2415

.60

± 19

3.89

ab24

.221

.41 *

*

3N

umbe

r of v

illi /

mm

24.

71 -

7.85

7.85

±

0.33

cd4.

71 -

7.85

6.12

±

0.37

cd4.

71 -

420.

565.

97 ±

0.

39c

3.14

- 6.

285.

65 ±

0.

42d

7.85

-10.

999.

42 ±

0.

41b

9.42

- 12

.56

11.3

0 ±

0.39

a21

.45

16.4

6 **

4N

umbe

r of G

oblet

ce

lls /

mm

247

.10

- 61

.23

54.6

4 ±

1.48

d47

.10

- 81.

6459

.66

± 3.

23c

61.2

3 - 7

2.22

67.0

4 ±

1.10

b47

.10

- 70.

6560

.45

± 2.

26c

54.9

5 - 6

5.94

61

.70

± 1.

24c

70.6

5 - 8

3.21

77.4

0 ±

1.19

a8.

4721

.52 *

*

5Th

ickne

ss o

f tun

ica

muc

osa

(µm

)77

4 - 1

485

762.

30 ±

52

.58c

1296

- 29

1620

93.4

0 ±

177.

24b

2088

- 25

5622

60.8

0 ±

52.9

0ab16

65 -

2700

2223

.90

± 12

0.71

ab15

48 -

2646

2160

±

121.

06ab

1629

- 31

9525

08.3

0 ±

188.

83a

18.8

514

.96 *

*

6Th

ickne

ss o

f lam

ina

prop

ria (µ

m)

9.00

- 27

.00

18 ±

1.9

0d18

- 36

25.2

0 ±

2.24

c36

- 45

39.6

0 ±

1.47

a18

- 27

21.6

0 ±

1.47

cd18

- 45

33.3

0 ±

2.70

b18

-36

25.2

0 ±

2.24

c22

.25

17.2

5 **

7Th

ickne

ss o

f m

uscu

laris

muc

osae

(

)

27 -

4534

.20

± 1.

80d

18 -

3625

.20

± 2.

24d

18 -

3627

.90

± 2.

49d

27 -

9063

± 8

.80c

63 -

9980

.10

± 3.

66b

72 -1

4496

.30

± 8.

17a

30.2

532

.82 *

*

8Th

ickne

ss o

f tun

ica

subm

ucos

a (µ

m)

18 -

2722

.50

± 1.

50c

18 -

2723

.40

± 1.

47c

18 -

3627

.90

± 2.

10bc

18 -

2723

.40

± 1.

99c

27 -

4533

.30

± 2.

34ab

27 -

4536

.90

± 2.

10a

22.8

8.89

**

9Th

ickne

ss o

f tun

ica

mus

cular

is (µ

m)

666

- 918

788.

40 ±

31

.36b

620

- 186

313

58 ±

16

1.58

a18

9 - 2

034

1431

±

211.

64a

675

- 176

412

42 ±

12

2.23

a63

0 - 2

610

1527

.30

± 21

2.70

a81

0 - 2

520

1670

.40

± 17

3.12

a37

.13.

79**

10Th

ickne

ss o

f tu

nica

sero

sa (µ

m)

45 -

108

77.4

0 ±

5.56

c72

- 14

411

6.10

±

7.64

bc13

5 - 1

9814

9.40

±

5.88

b90

- 16

211

6.10

±

7.03

bc11

7 - 6

3035

6.40

±

54.7

2a17

0 - 4

5037

1.70

±

17.2

9a37

.88

30.4

5 **

Star

t con

trol

gro

up

or G

roup

-IEn

d co

ntro

l gro

up o

r G

roup

-II

Tabl

e N

o. 5

: Sta

tistic

al a

naly

sis

of m

icro

met

rical

obs

erva

tions

of v

agin

a of

the

ovid

uct

of W

hite

Leg

horn

laye

rs (G

allu

s gal

lus d

omes

ticus

)

Sr.

No.

Para

met

erF'

Val

ueEn

d co

ntro

l gro

up o

r G

roup

-VSt

art c

ontr

ol g

roup

or

Gro

up-IV

You

ng h

ens

(32

wee

ks)

Spen

t hen

s (9

0 w

eeks

)Pr

een

glan

d ex

tirpa

ted

grou

p-III

%

of

CV

Pree

n gl

and

extir

pate

d gr

oup-

VI

NS

: Non

sign

ifica

nt,

*

: (P

d”

0.05

) an

d *

* : (

P d”

0.0

1)N

ote

: Mea

n w

ith a

t lea

st o

ne c

omm

on su

pers

crip

t do

not d

iffer

sign

ifica

ntly

39


dissertation submitted to the Council of the Collegeof Agriculture at Baghdad.

Khokhlov, R. YU. and Kuznetcov, S. I. (2007).Morphogenesis of a Tunica Mucosa of Oviduct of theHens. Int J. Morpho. 25 (2) : 329 – 333.

Kanchana R., Dhote, B. S. and Singh, G. K. (2006).Histological Studies on Shell Gland of Guinea Fowl(Numida meleagris) Technical Bulletin of XXIConvention of IAVA : 21

King, A. S. and McLelland, J. (1975). In “Outlines ofAvian Anatomy” Bailliere Tindall, London Pp : 65 –73.

Mehta S., Prasad, G. and Sahay, P. N. (2005). IndianJ. Vet. Anat. 17 (1 and 2) : 24 – 27.

Mohammadpour, A. A. and Keshtmandi, M. (2008).World J. Zoology 3 (2) : 47 – 50.

Ozen, A. E. Ergun and Kurum, A. (2009). J. AnkaraÜniv. Vet. Fak Derg. 56 : 177– 181.

Ray S., Chaudhary, M. N., Bhattacharya, B. andGhosh, R. K. (1971). Indian Vet. J. 71 : 39 – 43.

Richardson, K. C. (1935). In “The histology of thefowl”. Academic Press INC Ltd., 24-28 Oval Road,London Nw1 Pp : 354, 355 and 362.

Romanoff, A. L. and Romanoff, A. J. (1949). In “Thehistology of the fowl”. Academic Press INC Ltd., 24-28 Oval Road, London Nw1 Pp : 359, 361 and 371.

Saber A. S., Emara, S. A. M. and AboSaeda, O. M.M. (2010). Light, Scanning and Transmission ElectronMicro-scopical Study on the Oviduct of the Ostrich(Struthio camelus). J. Vet. Anat. 2 (2): 79 – 89.

Sandoz, D. (1971). In “The histology of the fowl”.Academic Press INC Ltd., 24-28 Oval Road, LondonNw1 : 367.

Sharma, R. K. and Duda, P. L. (1986). Generalhistology vis-à-vis the secretory phenomenon inmagnum of common Mallard. Indian J. Poultry Sci..21(1) : 49 – 52.

Sturkie, P. D. (1975). Avian Physiology, Third edition,Pp – 302 – 371.

Surface, F. M. (1912). In “The histology of the fowl”.Academic Press INC Ltd., 24 – 28 Oval Road,London Nw1 Pp : 352, 362 and 371.

Thomas, C. (2003). Female Reproductive System II :Avians, Veterinary Histology, Laboratory Exercises,Macbeth Act V. Sc. viii, lines 8 – 17.

Timo, J. N. (1969). Electron microscope observationson the shell gland mucosa of calcium-deficient hens(Gallus domesticus) Anatomical Record 164 (2) : 127– 139.

Wyburn G. M., Johnston, H. S. , Draper, H and MaidaF. Davidson (1973). The ultra structure of the shellforming region of the oviduct and the developmentof the shell of Gallus domesticus. ExperimentalPhysiology 58 :143 – 151.

40


Determination of phytate and bioavailability of iron from two varieties of legumes(Bengal gram-Cicer arietinum and kabulichana) and effect of processing on them

Kakar D.A.1,*, Udipi S.A.2, Ghugre P.S.2

1Department of Animal Nutrition, Bombay Veterinary College, Parel, Mumbai - 400 012, India2Department of Postgraduate Studies and Research in Home Science, SNDT Women’s University,

Juhu, Mumbai - 400 049, India *E-mail : [email protected]

ABSTRACT

Phytate is a constituent of grains and legumes. It is known to decrease the bioavailabilityof zinc, iron and other trace elements in humans. However, some processing is reported toreduce their phytate content. An in vitro method was used for determination of phytate andavailability of non-heme iron from two varieties of bengal gram, ie., Cicer arietinum (blackvariety) and kabulichana (white variety). Effect of processing/cooking on phytate and ironavailability was also investigated. Phytate content was found to reduce significantly onsoaking, dehulling, fermentation of the legumes and steaming. However, in case of punjabichole, though soaking reduced the phytate content, the content increased on preparation.This could be due to the addition of tamarind in the preparation. Processing/cooking wasfound to increase total available, soluble and ionizable iron in case of soaked, steamed andfermented legumes. Preparation of punjabi chole, however, increased the total availableiron to a lesser extent and significantly reduced the soluble and ionizable iron content.

Keywords: Phytate, Non-heme iron, Dehulling, Soaking, Fermentation, Pressure cooking

INTRODUCTION

Iron deficiency anaemia is a wide spreadnutritional problem affecting all segments ofpopulation in general, and children, women andpregnant women in particular. In the latter group,prevalence is as high as 60-70%. Among the factorsthat lead to iron deficiency, low availability is one ofthe most important factors, particularly so in the caseof pure vegetarians due to the poor bioavailability ofiron caused by the inhibitors such as phytates, tanninsand fibre in plant foods. Legumes and pulses bindcations like iron and zinc into mineral phytates. Theseare complexes of low solubility and therefore reduceavailability of these essential minerals.

Phytic acid (myoinositol 1, 2, 3, 4, 5, 6-hexakisdihydrogen phosphate) occurs primarily as a salt ofmonovalent and divalent cations in discrete regionsof cereal grains, legumes, some roots and tubers.Phytate also interacts with enzymes such as trypsin,

pepsin, alpha-amylase and beta-galactosidase,resulting in a decrease of their activity. It appears fromthe limited available literature that fermentation,dehulling, soaking and cooking could improve theavailability of iron. These increases correlated withreduction in phytates. Bengal gram, Cicer arietinum,has 5-6 mg of iron but it also has phytates which hinderiron absorption. Bengal gram is widely consumed allover India by all communities, using different recipes.The legume is generally dehulled, soaked, fermentedor cooked without fermentation. Changes inavailability of iron from these sources resulting fromprocessing needs to be studied.

The most reliable method for determiningbioavailability of iron from diets is to measure ironabsorption in humans using the extrinsic method assuggested by Cook et al. (1972), Layriss et al. (1974)and Hallberg et al. (1974). Though these invivomethods are accurate, they are, however, timeconsuming and expensive. An invitro method, on the

41


other hand, would have several advantages for rapidscreening and testing of availability of iron from diets.As most of the dietary iron is absorbed from the smallintestine and not from the stomach, this method alsodoes not determine the true availability of iron. At thealkaline pH of the intestine, solubility of iron decreasesconsiderably and at this alkaline pH the proportion ofionizable iron will be very low.


Six samples each of bengal gram (black variety)and kabulichana (white variety) with their variousprocessing treatments were analysed in triplicate fortheir phytate content and total available, soluble andionizable iron content using invitro methods.

Treatments given to samples for analysis :

a) Sample treatment for dry samples: Dry samplesof bengal gram, bengal gram dal (broken bengalgram) and kabulichana were examined for theirphytate and total, soluble and ionizable ironcontent. For this, 250 g of each sample wasground to a fine powder passed through 0.5mmsieve and stored in airtight containers till furtherprocessing.

b) Sample treatment to study the effect of soaking:Samples of bengal gram (black variety),kabulichana and bengal gram dal were soakedin distilled water for 12 hrs at 300C. Seed towater ratio used was 1:5 w/v. The soaked seedswere washed and rinsed with distilled water. Thesample was then ground to fine paste in a mixer.

c) Sample treatment to study the effect of dehulling:Bengal gram dal with skin was soaked overnightas described above and then hulls were removedmanually. The sample was then ground to finepaste in a mixer.

d) Sample treatment to study the effect offermentation: Dhokla was prepared to see theeffect of fermentation. For this standardprocedure as described in “Cooking Manual” ofS.V.T. College of Home-Science, Mumbai, wasfollowed. The samples were analysed at threestages of preparation: unfermented mixture ofthe batter, fermented mixture (10 hrs

fermentation) and steamed (10 min.) dhoklas.e) Sample treatment to study the effect of ordinary

cooking: Kabulichana was soaked for 12 hrs, putin a vessel surrounded by water in a pressurecooker and cooked for 15 minutes. Noingredients were added.

f) Sample treatment to study the effect ofpreparation of punjabi chole: Chole was preparedas per standard procedure as described in“Cooking Manual” of S.V.T. College of Home-Science, Mumbai

Preparation of processed samples : All theprocessed samples were dried in the hot air ovenkept at 600C to a constant weight, ground in anelectric grinder using a 0.5 mm sieve size andpacked in airtight containers for further chemicalanalysis.

Analytical method :

A. Phytate estimation : The sample extract washeated with an acidic iron-III solution of a knowniron content. The decrease in iron (determinedcolorimetrically with 2,2’ – bipyridine) in thesupernatant was considered as a measure for thephytate content. The standard was calibrated withstandard phytate solution for each set of analyses.

(i) Phytate reference solution: The sodium salt ofphytic acid, type V, 97% purity, containing about15% water, obtained from Sigma (No. P-5756)was used without further purification. Stocksolutions were prepared containing 0.15 gsodium phytate in 100 ml distilled water. Asphytate is absent, these stock solutions are stable.The reference solutions were prepared bydiluting the stock solutions with HCl in a rangefrom 3 to 30 mcg/ml phytate phosphorus (about0.2-11.7 ml stock solution in 100 ml). The finalHCl concentration in the reference solutions was0.2 N.

(ii) Ferric solution: Ammonium iron (III) sulphate12H2O (Merck Art 3776) 0.2 g was dissolved in100 ml 2N HCl and made upto 1000 ml withdistilled water.

(iii) 2,2’ Bipyridine solution: Ten grams of2,2’bipyridine (Merck Art 3098) and 10 mlthioglycollic acid (Merck Art 700) were

42


dissolved in distilled water and made upto 1000ml.(These solutions are stable for several months atroom temperature).Procedure : The phytate present in the sampleswas extracted by treating 0.5 g of the raw legumepowder with 20 ml of 0.2 N HCl and shaking inan incubator at 370C, 200 rev/min for 2hrs. Theextract was centrifuged at 8000 x g for 10 min at300C to obtain a clear supernatant. The extract(0.5 ml) was pipetted into a test tube fitted witha ground glass stopper. Ferric solution (ii), 1 mlwas added. The tubes were stoppered and fixedwith a clip.The tubes were heated in a boilingwater bath for 30 min. Care was taken for thefirst 5 min that the tubes remain well stoppered.After cooling in ice water for 15 min, the tubeswere brought to room temperature. Once thetubes reached room temperature, 2 ml of solution(iii) was added to the test tube and the contentswere mixed. The absorbance was measured aftera defined time (0.5-1 min) because the bipyridinereacts with the iron phytate and therefore thecolor changes with time (Haug and Lantzch,1983).

B. Total available, Soluble and Ionizable IronEstimation : This method is based on the releaseof ionizable iron from foods subjected totreatments with pepsin-HCl at pH 1.35 andsubsequent adjustment made to pH 7.5simulating somewhat conditions prevailing in thestomach and the intestine, respectively. Fordetermination of non-heme iron, food wasextracted with pepsin-HCl at pH 1.35 andsubsequently the pH was adjusted to 7.5 andfiltered. Ionizable iron was determined at pH 7.5using the 2,2’-dipyridyl method. Soluble ironwas determined as described by Tennet andGreenman (1969) by digesting the filtrate withpotassium permanganate which was colorizedwith ascorbic acid and filtered and the iron inthe filtrate was determined by 2,2’-dipyridylmethod. Total iron was determined on dry ashedsamples as described by AOAC (1995) and ironin mineral solution was estimated by thiocyanatemethod as reported by Wong (1965).

Treatment of foods : Dry foods were powderedand passed through a sieve of 9.25 mm meshsize and the powder used for determiningavailable iron. Soaked and cooked legumes(using standard method) were homogenized in ablender with deionised water to a creamyconsistency.

Extraction of soluble and ionizable iron : Drysample (2 g) was mixed with 25 ml of pepsin-HCl (0.5% pepsin in 0.1 N HCl). The pH of themixture was adjusted to 1.35 with distilled HCland incubated in a 100 ml conical flask at 370Cin a metabolic shaker waterbath for 90 min. Atthe end of this incubation, the contents of theflask were centrifuged at 3000 rpm for 45 minand the supernatant filtered through WhatmanNo. 44 filter paper. Soluble and ionizable ironwere determined in aliquots of the filtrate at pH1.35. In another flask pH was adjusted to 7.5with NaOH and incubated at 370C for 90 min inmetabolic shaker waterbath. At the end of thisincubation period the contents of the flask werecentrifuged at 3000 rpm for 45 min and thesupernatant was filtered again and the filtrate wasused for the determination of soluble andionizable iron

Analytical method :

(1) Total available iron : The food sample (2 g) wasdry ashed as described by AOAC (1995) and totaliron in the mineral solution was estimated by thethiocyanate method as reported by Wong (1965),where to an aliquote of 6.5 ml, 1.0 ml of 30%H2SO4, 1.0 ml potassium persulfate solution and1.5 ml of 40% KCNS solution are added. Thered colour that develops is measured within 20min at 540 nm. (Note: KCNS should be addedjust before taking the readings).

(2) Soluble and ionizable iron : Soluble iron wasdetermined as described by Tennet andGreenman (1969) by digesting the filtrate withpotassium permanganate which was colorizedwith ascorbic acid and filtered and iron in thefiltrate was determined by the alpha-alpha-dipyridyl method, where 0.6 ml of iron

43


containing solution was mixed with 1 ml of acidKMnO4 mixture (4 parts 3% w/v KMnO4 and 6parts N/2 HCl). It was then left at roomtemperature for 15 min. Ascorbic acid solution(0.4 ml 20%) was added and mixed. Iron wasthen estimated using method of Young and Hicks(1965) on Autoanalyser.

Ionizable iron : Free form of iron in the filtratewhich reacts with alpha-alpha-dipyridyl to yieldcolor retained after incubation of foods at pH1.35 and 7.5 was determined as described byAOAC (1995). Free form of iron corresponds toionizable iron (Shackleton and McCance, 1936).

Statistical analysis : The mean and standarddeviation of each parameter in each group werecalculated. Repeated measures analysis ofvariance (ANOVA) was carried out to assess thedifferences among various groups (Snedecor andCochran, 1994). All computations were carriedout using SPSS (version 11.5).

RESULTS AND DISCUSSSION

A. Phytate content :

a. Effect of soaking and dehulling on phytatecontent (Table -1 and 2) : Raw bengal gram dal(Cicer arietinum, broken with skin) was foundto contain 3.89 mcg/g of phytates. On soakingthe content was significantly (P< 0.01) reducedto 2.81 mcg/g. Dehulling the soaked grainsfurther reduced (P<0.01) the phytate content to1.44 mcg/g when compared with the soakedgrains. In case of Bengal gram whole the

reduction in phytate content was from 4.95 to1.98 mcg/g which was highly significant(P<0.01). Similarly, in case of kabulichana,phytate content decreased (P<0.01) from 2.96to 1.65 mcg/g.Duhan et al. (2002) observed similar effect onphytates due to soaking and dehulling. Theysubjected pigeon pea to various domesticprocessing methods like soaking and dehulling.The unprocessed seeds contained considerableamounts of phytic acid (857 mg/100 g). This wasreduced significantly (P<0.05) by 6-28% and30% in soaked and soaked-dehulled grains,respectively. Hotz and Gibson (2001) studied theeffect of soaking on maize flour and poundedmaize. They found that the treatment resulted in43 and 49% retention of phytate, respectively.

b. Effect of fermentation on phytate content(Table - 3) : Fermentation was found to reduce(P<0.01) the phytate content of dhokla batterfrom 1.93 in case of unfermented batter to 0.53mcg/g. and after steaming the content furtherreduced to 0.46 mcg/g. However, the differencein phytate content between fermented andsteamed dhoklas was found to be insignificant.Reddy and Pierson (1994) studied the effect offermentation and steaming on Indian foods likeidli, khaman and dhokla and found that 41.4, 58.2and 97.2% of the phytate gets hydrolysed,respectively, during natural fermentation.Rajalaxmi and Vanaja (1967) reported a smallerdecrease of 35% in phytate content of the Indianfermented foods, idli and khaman. Eka (1980)

Table 1. Phytates (mcg/gm) in Bengal gramwhole (Black Variety)Sample No. Dry Soaked

1 4.84 2.00

2 5.30 2.20

3 4.93 2.02

4 4.91 2.02

5 4.90 1.88

6 4.83 1.80

Table 2. Phytates (mcg/gm) in Bengal gram dalSample Dry with Soaking DehullingNo. skin with skin after

soaking1 4.00 3.18 1.20

2 4.00 2.60 1.87

3 3.82 2.60 1.10

4 4.02 2.84 1.80

5 3.82 2.84 1.58

6 3.70 2.84 1.10

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reported 39.2% reduction in phytate contentduring natural fermentation in the Africanfermented food, dawadawa.Antony and Chandra (1998) studied the effectof fermentation on antinutrient reduction andenhancement in availability of protein, starch andminerals in finger millet flour. They found thatphytate content decreased by 39% (P<0.05) by48 hrs. Phytate reduction has been reported inseveral fermented foods made from pearl millet(Dhanker and Chauhan, 1987), maize (Amoa andMuller, 1976), wheat (Gupta et al., 1992),sorghum (Marfo et al., 1990) and cereal-pulsemixtures (Chavan and Kadam, 1989).Hotz and Gibson (2001) assessed home-basedprocessing methods to reduce the phytatecontent of white maize. They found thatfermentation of cooked maize flour porridgeswith added germinated flour resulted in 54-86%retention of phytate compared to controls.Soaking maize flour or pounded maize anddecanting excess water resulted in 43 and 49%retention of phytate, respectively. Sripriya et al.(1996) found that germination and fermentationof finger millets reduced the phytate contentdecreased by 60%.Reddy and Pierson (1994) also studied the effectof fermentation on reduction in phytate inplant foods. In plant foods they found that innatural or pure mixed-culture fermentations byyeasts, molds and bacteria reduced the phytatein whole wheat breads upto 50% and in foodsmade from beans upto 95%. They observed thatthe highest rate of phytate hydrolysis occurredduring the proofing time; for brown and white

breads, baking and cooling further decreasedphytate content.Additionally, McKenzie-Parnell and Davies(1986) noticed little or no phytate hydrolysisin unleavened bran and whole meal breads. Lessthan 10% of the phytate was hydrolysed inunleavened whole meal bread. The maximumamount of phytate was hydrolysed in whitebreads. Hydrolysis of phytate was low whenwhole grain was fermented in the production ofkenkey, a sour fermented maize dumpling, andrabadi, (Amoa and Muller, 1976; Dhamker andChauhan, 1987). Hydrolysis of phytate occurredduring various stages of tempe preparation(Sutardi and Buckle, 1985; Paredes-Lapez andHarry, 1989; Nout and Rombouts, 1990). Garcia-Estipa et al. (1999) in their study on milled cerealproducts and breads found that loss of phytaterelative to unprocessed flours was between 20%for oat bread and 50% for white bread.

c. Effect of cooking on phytate content (Table -4): Soaking of kabulichana significantly(P<0.01) reduced phytate content from 2.95 to1.65 mcg/g. Steaming further reduced (P<0.01)phytates to 1.36 mcg/g which was significantlylower than the dry sample. However, preparationof punjabi chole increased (5.00 mcg/g) thephytate content significantly (P<0.01). Thiscould be due to addition of tamarind pulp whichwas added to impart sour taste and brown colorto the preparation.Reddy and Pierson (1994) also found that boilingand steaming results in a reduction in phytate

Table 3. Phytates (mcg/gm) in DhoklaSample Unfermented Fermented Steamed

No. batter batter dhokla1 1.84 0.48 0.56

2 2.24 1.00 0.70

3 1.40 0.30 0.20

4 2.00 0.70 0.70

5 2.00 0.40 0.40

6 2.08 0.30 0.20

Table 4. Phytates (mcg/gm) in Kabuli Chana(White Variety)

Sample Dry Soaked Steamed PunjabiNo. chole1 2.80 1.80 1.00 4.98

2 3.10 1.80 1.28 5.00

3 3.18 1.80 1.54 5.08

4 2.88 1.50 1.54 5.00

5 2.80 1.50 1.28 4.96

6 3.00 1.50 1.50 5.00

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in soyabeans but not in common beans. Agte etal. (1999) found that Indian vegetarian meals aresignificantly (P<0.01) higher in phytate contentthan African, Chinese and Western meals.Duhan et al. (2002) studied the effect of soakingand dehulling, ordinary as well as pressurecooking and germination on high yieldingcultivar of pigeon pea (Cajanus cajan). Theyfound that the high content of phytate (857 mg/100 g) in unprocessed seeds was reducedsignificantly (P<0.05) by 6-285, 30, 4-32, 4-36,35-45%, respectively, during soaking, soakingand dehulling, ordinary and pressure cooking andgermination.

B. Total, soluble and ionizable iron content(Table - 5) :

The total, soluble and ionizable iron at pH 1.35and 7.5 in the samples studied are given in theTable No. 5. At pH 1.35 the soluble iron wasslightly higher than the ionizable iron in all thesamples. When the pH was increased from 1.35to 7.5, both the ionizable and soluble irondecreased in all the samples, the decrease in theionizable iron, however, was of a greater

magnitude. The significant (P<0.01) reductionin soluble and ionizable iron at pH 7.5 ascompared to pH 1.35 may be due to reducedsolubility of iron at pH 7.5 which similates pHof intestine.It was observed that soaking and dehullingsignificantly (P<0.01) increased the totalavailable, soluble and ionizable iron content ofbengal gram (black variety) and chanadal at bothpH. Further comparison of treatment meansindicated that the increase was significantly(P<0.01) greater in case of dehulling.In case of dhokla, fermentation and steamingwere found to significantly (P<0.01) increasetotal available iron. Further comparison oftreatment means indicated that there was nostatistical difference in available iron betweenfermented and steamed dhokla. In case of solubleiron at pH 1.35 it was found that fermentationsignificantly (P<0.01) increased the content butsteaming reduced the content significantly(P<0.01). In case of ionizable iron, at same pH,it was found that fermentation and steaming hadsignificant (P<0.01) effect on it, with steamingshowing statistically higher (P<0.01) ionizableiron content as compared to fermented batter. At

Table 5. Total and percent soluble and ionizable iron content in raw and processed samples

Sr. Name of the Sample Total Iron pH 1.35 pH 7.5No. (mg %) Sol.Iron Ioni. Iron Sol.Iron Ioni.Iron1. Bengal gram (dry) 4.6 43.4 31.3 21.7 18.4

2. Bengal gram (soaked) 5.9 46.4 34.7 24.9 23.6

3. Chanadal (dry) 5.3 45.2 36.1 22.5 21.0

4. Chanadal (soaked) 6.5 49.7 40.5 23.4 22.5

5. Dhokla batter (unfermented) 6.4b 49.0b 41.0c 23.2c 22.1b

6. Dhokla batter (fermented) 6.8a 53.1a 49.7b 24.5a 22.9a

7. Dhokla batter (steamed) 6.7a 24.7c 52.3a 23.8b 21.0c

8. Kabuli chana (dry) 5.0c 38.2b 33.4b 20.3b 18.2b

9. Kabuli chana (soaked) 6.2a 47.6a 34.4a 25.7a 22.5a

10. Kabuli chana (steamed) 6.3a 47.8a 34.5a 25.9a 22.6a

11. Kabuli chana (punj. chole) 5.2b 35.2c 28.3c 19.2c 17.4c

* Values with same superscript in each row do not differ significantly (p<0.05)

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pH 7.5 soluble iron content significantly(P<0.01) increased due to fermentation andsteaming. However fermentation was found tohave greater effect on it. Fermentationsignificantly (P<0.01) increased the ionizableiron content at same pH whereas steaming wasfound to reduce it. Soaking and steaming ofkabuli chana were found to significantly(P<0.01) increase the total available iron thoughthe increase was found to be greater in case ofsteaming. In case of punjabi chole total availableiron increased but to a lesser extent compared tosoaking and steaming. Further, soaking andsteaming significantly (P<0.01) increased thesoluble and ionizable iron content at both pH,whereas preparation of punjabi cholesignificantly (P<0.01) reduced all the levels. Thissignificant reduction in the levels of iron couldbe due to presence of mineral binding agents(tannin) in tamarind which was used to givesourness to the product.Duhan et al. (2002) in their study on HClextractability of Ca, P and Fe as affected byvarious domestic processing and cookingmethods found that HCl extractability of dietaryessential minerals like total Ca, P and Fe wasenhanced significantly when the pigeon peaseeds were processed and cooked, possibly dueto reduction in phytate content, which is knownto chelate the minerals.Antony and Chandra (1998) in their study onthe effect of fermentation on antinutrientreduction and enhancement in mineralavailability in finger millet flour found that HClextractability of minerals increased significantlyand that of iron doubled. The extractability ofminerals (Ca, P, Fe and Zn) showed a significantnegative correlation (P<0.05) with the phytatelevels.Sripriya et al. (1996) found increase in HCl-extractable minerals of 47%. They observed thatCa, P and Fe extractability increased from 162,49.1, 0.34 mg/100 g to 181, 55.3, 0.49 mg/100 gon germination which further increased to 224,107 and 1.33 mg/100 g, respectively, onfermentation (24 hrs).Fermentation was more effective in increasingthe bioavailability of Ca, P and Fe. Iron

availability from finger millet improved from0.34 to 1.4 mg/100 g due to processing.Annapurani and Murthy (1985) in their study onthe effect of dehulling, milling, roasting andcooking on total and absolute iron of selectedtraditionally used pulses and found thepercentage total and absolute available iron (mg/100 g) content of dehulled and milled pulses –bengal gram (10, 156) increased whereas roastedand cooked bengal gram showed percentdecrease of total iron (7 and 23) and percentincrease of absolute available iron (124 and 92),respectively.Lee and Clydesdale (1979) have stated that foodprocessing such as germination, roasting,cooking and milling showed two or four foldincrease in biologically available iron. Satwadharet al. (1981) proved that the resultant significantincrease in absolute available iron (20 to 50%)is also due to decrease in polyphenols aftercooking.

CONCLUSIONA study was conducted to see the effect of

processing and cooking on phytate and availabilityof non-heme iron on two varieties of bengal gram –Cicer arietinum (black variety) and kabulichana(white variety) using invitro method. It was found thatphytate content reduced significantly when thelegumes were soaked, dehulled, fermented andpressure cooked but the phytate content increasedsignificantly when punjabi chole was prepared. Thiscould be due to addition of tamarind in the preparation.Soaking, dehulling and fermentation was also foundto increase total available iron, soluble and ionizableiron content of the legumes. However, preparation ofpunjabi chole did not seem to increase the same.

Further studies are suggested to see whetheraddition of pomegranate seeds or mango powder assouring agent is preferable to tamarind for improvingthe total, soluble and ionizable iron and reducing thephytate content.

REFERENCES

Agte V.V.; Tarwadi K.V. and Chiplonkar S.A. (1999)J of Food Composition and Analysis, 12:161-167.

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Amoa B. and Muller H.G. (1976) Cereal Chem,53:365-375.

Annapurani S. and Murthy N.K. (1985) The Ind J NutrDietet, 24:95-105.

Antony U. and Chandra T.S. (1998) J Agri FoodChem, 46:2578-2582.

AOAC (1995). Official methods of analysis. 16th edn.Association of Official Analytical Chemists,Washington DC.

Chavan J.K. and Kadam S.S. (1989) CRC Crit RevFood Sci Nutr, 28:349-400.

Cook J.D., Layriss M.; Martinez-Torres C.; WalkerR.; Monsen E. and Finch C.A.(1972) Clin Invest, 51:805.

Dhanker N. and Chauhan B.M. (1987) J Food Sci,52:828-829.

Duhan A.; Drago S.R. and Valencia M.E. (2002) J ofFood Sci, 7(8):3130-3134.

Eka O.U. (1980) Food Chemistry, 5: 303-308.

Garcia-Estipa R.M.; Guerra-Hernandez E. and Garcia-Villanova B. (1999) Food Research International,32:217-221.

Gupta M.; Khetarpaul N. and Chauhan B.M. (1992)Plant Foods Hum Nutr, 42:109-116.

Hallberg L.; Garby L.; Suwanik R. and Bjorn-Rasmussen E. (1974) Am J Clin Nutr, 27: 89.

Haug W., and Lantzch H.J. (1983) I Sci Food Agri,34:1423-1426.

Hotz C. and Gibson R.S. (2001) J Agri Food Chem,49:632- 698.

Layriss M.; Martinez-Torres C. and Gonzalez K.(1974) Am J Clin Nutr, 27: 152.

Lee K. and Clydesdale F.M. (1979) CRC Crit RevFood Sci Nutr, 11:117.

Marfo E.K.; Simpson B.K.S.; Idowu J. and Oke O.L.(1990) J Agri Food Chem, 38:1580-1585.

Mckenzie-Parnell J.M. and Davies N.T. (1986) FoodChem, 22:181-192.

Nout M.J.R. and Rombouts F.M. (1990) J ApplBacteriol, 69:609-633.

Paredes-Lopez O. and Harry G.I. (1989) L Food Sci,54:968-970.

Rajalaxmi R. and Vanaja K. (1967) Br J Nutr, 2:467-473.

Shackleton L. and McCance R.A. (1936) Biochem J,30:5.

Snedecor C.W. and Cochran (1994) Statisticalmethod, 6th Edition, Iowa State University Press,U.S.A.

Sripriya G.; Antony U. and Chandra T.S. (1996) FoodChem, 58(4):345-350.

Sutardi and Buckle K.A. (1985) J Food Sci, 50:260-263.

Tennet G.B. and Greenman D.A. (1969) J Clin Path,22:301-303.

Wong (1965) Quoted in Oser B.L. - Hawk’sphysiological chemistry 1965. 14th edn. McGrawHill Book Co. New York. p 1094-1095.

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Histomorphological Study on Laparoscopically Excised Ovaries in Dog (Canis domesticus)

Aware N.N., Shankhapal V.D., Dhande P.L., Khandekar G.S. and Kolekar V.P.Department of Veterinary Anatomy and Histology

Bombay Veterinary College, Parel, Mumbai - 400 012, India.

ABSTRACT

The present study was undertaken to study the gross anatomical and histological structureof ovaries in dog excised by laparoscopy as well as to evaluate the most feasible approachfor performing laparoscopic ovariectomy in dog. Ovaries of 24 dogs were removed bylaparoscopic ovariectomy from the clinical trials conducted in the department of VeterinarySurgery and Radiology, Bombay Veterinary College, Parel, Mumbai. The ovaries werecollected in 10% neutral buffer formalin solution for its histological study. The grossanatomical and histological study was carried out in the department of Veterinary Anatomyand Histology, Bombay Veterinary College, Parel, Mumbai. In laparoscopic view, the ovarieswere present towards the cranial end of the urinary bladder and dorsal to it enclosed in theovarian bursa. The ovaries were reddish pink in colour. The size of ovaries were variabledepending on the phase of estrus cycle. Numerous developing follicles at various stages ofdevelopment were present in the cortical stroma. The stroma of the ovary showed extensivehemorrhagic areas. The histochemical and histoenzymic activities varied between the differentcomponents of ovaries. The diameter of graafian follicle and the diameter of ovum in boththe ovaries were increased in the estrus stage. The laparoscopic ovariectomy was most feasiblewhen the dog were placed in 100 tredelenburg (head down) position in dorsal recumbency.

Keywords: Laparoscopy, Ovary, Dog, Histology

INTRODUCTION

The ovary constitutes to the structure of primaryimportance in the female reproductive system byrhythmically developing in the female germ cellsduring various stages of oestrous cycle and producingthe hormones which are essential for the sexual lifeof female. It performs both exocrine function asproduction of ova and endocrine function asproduction of ovarian hormones chiefly progesteroneand oestrogen (Priedkalns, 1993).The knowledge ofmorphological and histological structure of ovarieswill be useful for its physiological functions and alsofor the identification of critical periods of genitalorgan.

The intact female dogs are more prone to clinicalconditions like ovarian tumours and pyometra.

Behavioural changes and pseudo-pregnancy conditionalso arises in pet and stray dogs. So to treat suchclinical conditions, it is important to performovariectomy and ovariohysterectomy.

The advantages of laparoscopic surgery areminimal bleeding, decrease in post-operative stressand pain, faster recovery time, minimal scarring.Laparoscopic ovariectomy, laparoscopicovariohysterectomy and laparoscopic-assistedovariohysterectomy have been evaluated in bitchessince 1985 as neutering methods. (Wildt and Lawler,1985). Various laparoscopic surgical approach andstudy of anatomy of abdomen in llamas were carriedout. (Anderson et al., 1996).Single portal and twoportal laparoscopy for ovariectomy on dogs has beencarried out, both these methods being suitable forperforming laparoscopic ovariectomy in dogs. But

49


single portal access was more feasible as it allowedworking within the same plane of laparoscope, greatercontrol of instruments and obviating the need of asurgical assistant. (Kulkarni, 2012).Thus, a need arisesto study the most feasible approach of laparoscopicovariectomy, so as to reduce the operating time andto reduce the stress during surgery both for the patientas well as the operating surgeon. The knowledge ofmorphological and histological structure of ovarieswill be useful for its physiological functions and alsofor the identification of critical periods of genitalorgan. Hence, the present investigation is undertakento study the gross anatomical and histological structureof ovaries in dog excised by laparoscopy as well as toevaluate the most feasible approach for performinglaparoscopic ovariectomy in dog.


Ovaries of 24 dogs were removed bylaparoscopic ovariectomy from the clinical trialsconducted in the department of Veterinary Surgeryand Radiology, Bombay Veterinary College, Parel,Mumbai. The ovaries were immediately collected in10% neutral buffer formalin solution for itshistological study. The ovaries were categorizedaccording to proestrus, estrus, metestrus and diestrusphases of estrus cycle on the basis of ovarian structureThe gross anatomical and histological study wascarried out in the department of Veterinary Anatomyand Histology, Bombay Veterinary College, Parel,Mumbai.

The laparoscopic procedure for ovariectomyinvolved the placement of dog in 10 Ú tredelenburg(head down) position in dorsal recumbency. Thelaparoscope was inserted caudal to the umbilicus onventral midline incision. The two working port wereinserted through the right and left flank incisions,respectively, at equidistance from the laparoscope port(Fig.1). This provided a clearer, complete andthorough understanding of laparoscopic anatomy ofa dorsal recumbent animal. The broad ligament wasextended from the caudal border of the kidney andhad the uterine vessels. The ovarian ligaments andovaries were observed on elevation of the uterine horn.The bipolar electrocautery was used for coagulationand ligation during laparoscopy. By performinglaparoscopic ovariectomy, ovaries were collected for

further study.The gross anatomical observations of the ovaries

were recorded in respect of shape, size, color andlocation laparoscopically. For the histological study,ovaries were fixed in 10 % neutral buffered formalin.About 3 – 5 mm tissue pieces from different regionsof each ovary were taken. These tissue pieces werethen treated with routine method of dehydration inascending grades of alcohol (ethanol), cleared inxylene and embedded in paraffin wax as per methodof Singh and Sulochana (1996-97).The paraffin blockswere sectioned at 3 to 5 µ thickness. The preparedslides were stained with various stains to make thehistological observations as described by Singh andSulochana (1996-97).Slides were stained withMayer’s Hematoxylin and Eosin stain for generalhistological observation, Van Gieson’s stain forcollagen fibers, Weigert’s stain for elastic fiber,Wilder’s stain for reticular fiber and Mallory’s triplestain for connective tissue. Histochemical reactionswere observed for the study of correlative changes inthe carbohydrate, lipids and enzymes in the ovariesby using various staining techniques. Slides werestained with Periodic Acid Schiff reaction (pH 2.5)stain for mucopolysaccharides (CHO) (Culling,1969),PAS Alcian blue (pH 2.5) stain for neutralmucopolysaccharides (Culling, 1969),Best Carminemethod for glycogen (Luna, 1968), Gomori’smodified alkaline phosphate cobalt method for

Fig. 1 : Photograph showing measurement of distance fromA: laparoscope port to B: right working port and C: leftworking port

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alkaline phosphatase activity at pH 8.5 - 9.5 (Singhand Sulochana, 1996 - 97) and Gomori’s modifiedlead nitrate method for Acid phosphate activity at pH5 (Singh and Sulochana, 1996 - 97)

RESULT AND DISCUSSION

In laparoscopic view, both the ovaries werefound dorsal to the bladder enclosed in the ovarianbursa. (Fig. 2) The ovaries were found in theabdominal cavity. Most of the ovarian structures likecorpus luteum, primary follicle and graafian folliclewere not distinguishable. It may be due to use ofelectrocautery leading to sealing of blood vessels byfusion of the lumen of blood vessels and leading to abursting pressure. Similar findings were reported byMassarweh et al. (2006). During laparoscopic study,the ovaries, oviduct, uterine horn, broad ligament werefound in dorsally recumbent position.

During gross anatomical study, both the ovariesof dog were found oval in shape. These observationsare in accordance with Ghosh (2006) in dog. Theovaries were found reddish pink in colour. However,

Nimunkar (1999) noted light pink colour of ovariesin ewes. This might be due to species variation andmethod of excision of ovaries. The ovaries presentedtwo surfaces, two borders and two ends. The lateraland medial surfaces were convex and irregular inappearance. The anterior end was tubal end and

attached to the fimbria of the infundibulum. Theposterior end was narrow and rounded. The right ovaryand right oviduct as well as the broad ligament wereobserved with the right ovarian artery. The left ovaryand left oviduct were observed with the broad ligamentand the left ovarian artery. The ovaries were caudallyattached to the uterus by the broad ligament. Theovarian ligaments were observed on elevation of theuterine horns. These observations are in accordancewith Culp et al. (2009) in dog, Sali (2010) in awassiewes and Khandekar (2011) in dog.

Histologically, both the ovaries were divided intocortex and medulla in all phases of estrus cycle. Inthe medulla of both the ovaries the network of looseconnective tissue was seen along with blood vesselsin all phases of estrus cycle. Similar observations werenoted by Eurell and Frappier (2006) and Nimunkar(1999) in sheep. The cortex of both the ovaries wascomposed of germinal epithelium, tunica albugenia,stroma having primary follicles and interstitial glands,graafian follicle, corpus luteum and ovum. (Fig. 3).The stroma of the ovary showed extensivehemorrhagic areas. This might be due to use ofelectrocautery use for ligation during laparoscopy.Massarweh et al. (2006) reported that the bipolarelectrocautery leads to coagulation by fusion ofintimal layers of vascular structure leading toobliteration of blood vessels and a bursting pressure.

Fig. 3 : Microphotograph of left ovary of dog in estrus stage(arrow) showing graafian follicle and (arrowhead) ovum.(HE, 100X)

Fig. 2 : Photograph of position of uterine hornslaparoscopically. A- Body of uterus, B- horns of uterus, C-urinary bladder D-Ovary

51


Germinal epithelium was found to be outermostcovering of the ovary. It was composed of simplesquamous or cuboidal epithelium. Joshi et al. (1977)observed that the germinal epithelium of goat ovarywas lined by simple cuboidal cells. This observationis in accordance with the findings of Bacha and Bacha(2000) in dog and Joshi et al (1977) in goat. In thepresent study, there was no difference in the germinalepithelium between right and left ovary in all phasesof estrus cycle. Tunica albugenia was found beneaththe germinal epithelium. It was composed of denseconnective tissue. Tunica albugenia was disrupted bythe growth of ovarian follicles and corpus luteumduring different phases of estrus cycle. Thisobservation is in accordance with the findings ofPriedkalns (1993) in dog. No difference was observedin the tunica albugenia between right and left ovaryin all phases of estrus cycle.

Primary follicles were present in the stroma ofthe ovary beneath the tunica albugenia. The primaryfollicles were composed of primary oocyte surroundedby simple cuboidal epithelium of follicular cells. Theprimary follicles gave rise to the graafian follicles.This observation is in agreement with the observationsreported by Bacha and Bacha (2000) in dog andPriedkalns (1993) in dog. There was no differencefound in the structure of primary follicles betweenright and left ovary in all phases of estrus cycle.Graafian follicles were the mature follicles surroundedby a sheath of stromal cells called the theca externaand theca interna cells. The graafian follicle composedof a fluid-filled antrum and tertiary oocyte. The oocytewas surrounded by a multilayer of membranagranulosa cells called the cumulus oophorus. Similarobservations are reported by the Bacha and Bacha(2000) in dog and Priedkalns (1993) in dog .Therewas no difference observed in the graafian folliclesbetween right and left ovary in all phases of estruscycle. In both the ovaries, regressions of some follicleswere found. In these follicles, ovum was founddegenerated. The ovum was observed in the maturegraafian follicle. The ovum was surrounded by a thickglycoprotein layer, the zona pellucida. The coronaradiata, a layer of columnar cells, was surrounded thezona pellucida (Fig. 4). These observations are inagreement with the Priedkalns (1993). There was nodifference noted in the structure of ovum between rightand left ovary in all phases of estrus cycle. Corpus

luteum was formed by the multiplied andhypertrophied cells of theca interna and the granulosacells. Similar observations were reported by the Bachaand Bacha (2000) in dog and Budras et al. (2007) indog. There was no difference observed in the structureof corpus luteum between right and left ovary in all

phases of estrus cycle. In all phases of estrus cycle,prominent interstitial glands were seen in the stromaof both the ovaries. Similar observations were notedby Bacha and Bacha (2000) in dog and by Eurell andFrappier (2006) in dog.

In both the ovaries, a dense network of reticularfibers was observed throughout the stroma of theovary, in the interstitial glands and in the medulla. Inboth the ovaries, the network of collagen fibers wereobserved in the stroma and the theca layerssurrounding the follicles. In both the ovaries, collagenfibers were predominated in the tunica albuginea,stroma and the theca layers surrounding the folliclesof the ovaries (Fig. 5 & 6). In both the ovaries theelastic fibers were observed in the medulla, in the wallsof blood vessels and in the fibrous layer of the thecalayers surrounding the follicles in all phases of estruscycle.

Mucopolysaccharide activity was found intensein the theca externa layer surrounding the follicle andthe zona pellucida of the ovum and moderate activity

Fig. 4 : Microphotograph of ovary in dog showing A: coronaradiata, B: granulosa cells, C: ovum, zona pellucida (arrow)and nucleus (arrowhead). (HE, 400X)

52


was observed in stroma, whereas weak activity wasobserved in the cortex of the ovary. This indicatesthat the presence of mucopolysaccharide in differentcomponent of ovaries varies in concentration. Therewas no difference for histochemical and histoenzymic

in right and left ovaries. Similar findings wereobserved by Das and Ghosh (2011) in adult mice. Theactivity of the neutral mucopolysaccharide wasmoderate in stroma of the ovary whereas it was intensein the zona pellucida and in the theca externa, whichindicated that the concentration of neutralmucopolysaccharide more. The tunica albugenia andcortex of the ovary showed low activity of neutralmucopolysaccharide. There was no difference inhistochemical and histoenzymic reaction in right andleft ovaries. The glycogen particle was present weaklyin the tunica albuginea. The glycogen particle wasfound moderate in stroma. The alkaline phosphataseenzymes activity was intense in the theca externa cells.The alkaline phosphatase activity was not found thein tunica albuginea, and in the stroma. The acidphosphatase enzymes activity was not seen in ovaryat different phases of estrus cycle.

In conclusion, in laparoscopic view, the ovarieswere present towards the cranial end of the urinarybladder and dorsal to it enclosed in the ovarian bursa.Numerous developing follicles at various stages ofdevelopment were present in the cortical stroma. The

Fig. 5 : Microphotograph of right ovary of dog arrowshowing collagen fibers. (MTS stain, 100X)

Fig. 6 : Microphotograph of right ovary of dog arrowshowing PAS activity. (PAS stain, 100X)

stroma of the ovary showed extensive hemorrhagicareas. The histochemical and histoenzymic activitiesvaried between the different components of ovaries.The laparoscopic ovariectomy was most feasible when

the dog were placed in 100 tredelenburg (head down)position in dorsal recumbency. While the laparoscopewas inserted caudal to the umbilicus on ventral midlineincision and the two working port were insertedthrough the right and left flank incisions, respectively,at equidistance from the laparoscope port.

REFERENCES

Anderson D. E., E. M. Gaughan, A. N. Baird, H. C.Lin and D. G. Pugh (1996). J. Am. Vet. Med. Assoc.208 (1): 111 – 6.

Bacha, W. J. and L. M. Bacha (2000). Textbook ofColour Atlas of Vet. Histology 2ndEdn, Pp. 221 – 227.

Budras, K. D., P. H. McCarthy, W. Fricke, R. Richter,A. Horowtiz and R. Berg (2007). Textbook ‘Anatomyof the dog’ 5th Edn. Budras (ed.): 66-67.

Culling, C. F. A. (1969). Handbook ofHistopatological technique (including museumtechnique).” 2nd Edn. Butter Worth and Co.,

53


Philadelphia and London, Pp. 228 – 238 and 477 –479.

Culp W. T. N., P. D. Mayhem and D. C. Brown (2009).Veterinary surgery, 38: 811-817.

Das, R. S. and S. K. Ghosh (2011). Nepal Med. Coll.J. 13(2): 77 – 83.

Eurell, J. A. and Frappier, B. L. (2006). Dellmann’stextbook of Veterinary histology’, 6th Edn. Blackwellpublishing, USA, 256-262.

Ghosh, R.K. (2006). Primary Veterinary anatomy’4thEdn, 246-248.

Joshi C. L., B. S. Nanda and R. P. Saigal (1977). Anat.Anz. 141(1): 19-36.

Khandekar, G. S (2011). Ph.D. Thesis submitted toMaharashtra Animal and Fishery Science University.Nagpur.

Kulkarni, M. V. (2012). M. V. Sc. Thesis submitted toMaharashtra Animal and Fishery Science University.Nagpur.

Luna, L. G. (1968). Histological Staining Methods ofArmed Forces Institute of Pathology, McGraw Hill,New York, NY, USA, 3rd Edn.

Massarweh, N. N., N. Cosgriff and D. P. Slakey(2006). J. Am. Coll. Surg. 202(3): 520-530.

Nimunkar, S. N. (1999). M. V. Sc. Thesis submittedto Maharashtra Animal and Fishery ScienceUniversity. Nagpur.

Priedkalns, J. (1993). Textbook of Vet Histology” 4th

Edn. Dellmann, H. D., Lea and Febiger, Philadelphia;233-241.

Sali, K. M (2010). J. Vet. Anat., 3(2): 47-54.

Singh, U. B. and S. Sulochana (1996–97). Hand bookof Histological and Histochemical techniques”Premier Publishing House, Hyderabad 110 – 111.

Wildt, D. E. and D. F. Lawler (1985). Am. J. Vet. Res.46(4): 864 – 869.

54


Intra-operative evaluation of intestinal viability in cases of obstruction in dog

Jadhav P.T., S.V. Upadhye, M.S. Dhakate and B.M. Gahlod,Department of Veterinary, Surgery & radiology, Nagpur Veterinary College, Nagpur,

Maharashtra Animal & Fishery Sciences University, Nagpur.

ABSTRACT

The Intra-operative evaluation of intestinal viability in cases of obstruction wasundertaken on nine clinical cases of intestinal obstruction in dogs presented during November2009 to June 2010 at Teaching Veterinary Clinical Complex, Nagpur Veterinary College,Nagpur. In cases of intestinal obstructions which required enterectomy, during the procedure,the assessment of intestinal viability by fluroscein sodium method is carried out by injectingfluroscein sodium @ 25 mg/Kg body weight IV. Appearance of dye in the blood vessels andits spread all along the intestinal segment was examined under darkened Operation theatreunder ultraviolet light. Clinically, the viability of the intestine was decided on the basis ofcolour of affected intestinal segment, intestinal peristalsis, mesenteric pulsation. Out of thenine cases of intussusceptions, five cases showed viability of the segments and four caseswere apparently non-viable when judged by clinical criteria. However, on fluorescein dyestudies, seven cases were observed to have proper blood supply and only two cases revealedcompromised viability. No adverse reactions to fluorescein dye were observed intra- orpostoperatively.

Keywords: Intestinal viability, Dog, Fluroscein dye

INTRODUCTION

Surgical intervention for intestinal obstruction isvery often required in the pet practice. The obstructioncould be caused due to intussusceptions, volvulus,torsion, lodgment of foreign body, or even intraluminalneoplasia. Long standing cases of obstruction maysuffer with venous drainage and arterial occlusionleading to ischemia followed by necrosis of the bowel(Levitt and Bauer, 1992). An accurate and timelydiagnosis and surgical relief of the obstruction isnecessary. The success of surgery depends on intestinalsegment viability. Therefore intraoperative use offluroscein sodium dye is gaining popularity. Therefore,the work was planned to study the efficacy of thismodality in diagnosing the viability of the intestinalsegment.


The study was carried out on nine clinical cases

of intestinal obstruction in dogs presented duringNovember 2009 to June 2010 at Teaching VeterinaryClinical Complex, Nagpur Veterinary College, Nagpur.

In cases of intestinal obstructions which requiredenterectomy, during the procedure, viability of theintestine was decided on the basis of colour of affectedintestinal segment, intestinal peristalsis, mesentericpulsation and fluroscein sodium study.

The exteriorized intestinal segment suspected forischemia was examined under long wave ultravioletlamp. The segment with pink colour, having arterialpulsation and peristaltic movement was taken as viable,whereas, segment with blue colour, lacking pulsationand peristaltic movements were taken as non-viable.The assessment of intestinal viability by flurosceinsodium method is carried out by injecting flurosceinsodium (10 percent solution) @ 25 mg/Kg body weightslow IV. Immediately after administration of injectionfluroscein sodium1, the operation theatre was darkened.Appearance of dye in the blood vessels and its spreadall along the intestinal segment was considered as

55


viable segment and non-appearance of florescence inthe segment in question was considered as non-viablesegment.


Assessment of intestinal viability was carried outusing two different criteria viz. clinical assessmentand use of intravenous fluorescein dye sodium. Theclinical criteria for assessing the viability of intestinalloop were colour of the intestinal loop, arterialpulsation and peristaltic movements. Pink colour,having arterial pulsation and peristaltic movementswere considered as viable (Fig. 1). In case offluorescent study, the exteriorized segment of intestinewas examined under long wave ultraviolet light. Outof the nine cases of intussusceptions, five casesshowed viability of the segments and four cases wereapparently non-viable when judged by clinical criteria.However, on fluorescein dye studies, seven cases wereobserved to have proper blood supply (Fig. 2) andonly two cases revealed compromised viability (Fig.3). No adverse reactions to fluorescein dye wereobserved intra- or postoperatively.

Acute intestinal ischemia and infarction remainserious clinical problems despite early operativeintervention. Accurate intraoperative assessment ofintestinal viability is essential in determining the limitsof resection in patients with intestinal infarction or inpatients wherein intestinal resection is required dueto intestinal damage of varying extent. In most of thecases, surgeon considered the return of colour, arterial

pulsation and visible peristalsis as the major basesfor decision. However, clinical features of bowelviability such as color and peristalsis do not correlateuniformly with bowel survival, and as a result, severaltechniques have been developed to assess intestinalblood flow at the time of operation. A false-negativeresult leaves in situ nonviable bowel, which may leadto early perforation and late stricture formation. Thissituation may be recoverable with further surgicalintervention, however, on the other hand, a false-positive assessment of bowel viability results in theresection of potentially recoverable intestine, whichis lost forever and may represent a vital difference formorbidity-mortality and long-term nutrition (Horganand Gorey, 1992). The surgeon has to decide theviability of the intestinal segment that has receivedinsult as a result of obstruction due to intussusception.In the clinical criteria, the segment with pink colour,having arterial pulsation and peristaltic movement wastaken as viable, whereas, segment with blue colour,lacking pulsation and peristaltic movements weretaken as non-viable.

In the present investigation, the fluoresceinSodium dye was administered @ 25mg/Kg. bodyweight as suggested by Tank et al. (2002) who opinedthat the trend and the quality of the intestinalfluorescence provided by the 25mg/kg i/v dose of thedye was adequate for getting interpretable intestinalfluorescence in dogs. Out of the nine cases ofintussusceptions, five cases showed viability of thesegments and four cases were apparently non-viablewhen judged by clinical criteria. However, on

Fig. 1 : Viable segment of intestine Fig. 2 : Fluorescein sodium dye study showing viableintestinal segments

56


fluorescein dye studies, seven cases were observedto have proper blood supply and only two casesrevealed compromised viability. Thus fluoresceinsodium dye studies seemed to be more reliablediagnostic tool as compared to standard clinicalevaluation. The, excess vital intestinal segment whichotherwise could have been removed, was saved.Further, there was no adverse or anaphylactic reactionin any of the case investigated. The fluorescein dyetechnique was found to be sensitive, specific,predictive and significantly more accurate than thestandard clinical criteria for intraoperative predictionof the small intestinal viability in dogs. Thus, it wasopined that fluroscein technique is the method ofchoice for the prediction of small intestinal recoveryfollowing ischemic injury caused due to compromisedblood supply as a result of intussusception.

Bulkley et al (1981) also observed that thestandard clinical judgment was moderately accurate(89%) but it led to relatively high rate (46%) ofunnecessary bowel resection. The fluoresceinultraviolet fluorescence pattern was correct in all 54determinant bowel segments and proved moresensitive, specific, predictive and significantly moreaccurate overall than the standard clinical judgment.

Similar observations have been noted by Dyess et al.(1991), Holmes et al. (1993), Kumar et al (1999), andTank et al. (2003).

REFERENCES

Bulkley, G. B., G. Zuidema, S. Hamilton., C. O’mara,P. G. Klacsmann and S. D. Horn (1981). Intraoperativedetermination of small intestinal viability followingischemic injury. Ann. Surg. 193(5):628-635.

Dyess, D. L., B. W. Bruner., C. A. Donnell., J. J.Ferrara and R. W. Powell (1991). Intraoperativeevaluation of intestinal ischemia: A comparison ofmethods. South Med. J. 84(8):966-974.

Holmes, N. J., G. Cazi., M. T. Reddell., J. H. Gorman.,B. Fedorciw., J. L. Semmlow and R. E. Brolin (1993).Intraoperative assessment of bowel viability. J.Investigative Surg. 6(2):211-221.

Horgan, P. G. and T. F. Gorey (1992). Operativeassessment of intestinal viability. Surg. Clin. NorthAm. 72(1):143-155.

Kumar, D. D., K. Ameerjan and W. P. Archibald David(1999). Pulorus and ileum viability assessement indogs. . Indian J. Vet. Surg. 20(1):37-38.

Levitt, L. and M. S. Bauer (1992). Intussusception indogs and cats: A review of thirty-six cases. Can Vet.J., 33:660-664.

Tank, P. H., N. N. Balasubramaniam and W. P.Archibald David (2002). Effect of dosase offluorescein sodium on trend and quality of intestinalfliuorescein in dogs. Indian J. Vet. Surg. 23(1):47-48.

Tank, P. H., N. N. Balasubramanian and W. P.Archibald David (2003). Assessment of standardclinical criteria and fluorescein dye technique forintraoperative prediction of small intestinal viabilityin dogs. Indian J. Vet. Surg. 24(2):83-88.

Fig. 3 : Fluorescein sodium dye study showing viable andnon viable intestinal segments

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Blood serum biochemical profile of WLH layers before and after uropygealectomy.

Patil A. D, Dhande P.L, Deshmukh B. T, Bakshi S. A, Ranade A.S and Ingole S.D.Department of Veterinary Anatomy and Histology,

Bombay Veterinary college, Parel, Mumbai - 400 012.

ABSTRACT

After removal of preen gland, the biochemical constituents of the blood serum of WLHyoung and spent hens were studied. Young hens aged 32 weeks (50 birds) and spent hens of90 weeks (50 birds) were subdivided into three groups each consisting of start control group(10 birds), end control group (20 birds) and preen gland extirpated group (20 birds).The serum biochemical constituents like calcium, phosphorous, triglycerides and totalcholesterol were increased as the age advanced. Further, the concentration of thesebiochemical constituents was found higher in preen gland extirpated groups than in theirrespective controls.

Keywords: Serum biochemical constituents, total cholesterol, triglycerides, calcium,phosphorous and uropygealectomy.

INTRODUCTION

Uropygeal or preen gland is only one pear shapedholocrine, cutaneous, oil secreting gland situated atthe base of tail or coccygeal vertebra which is welldeveloped in aquatic birds. Many workers reportedthat the secretion of the preen gland is closely relatedto hydrophobic property, which would have anessential role in plumage waterproofing. It is alsopossible that the gland could play a role in plumagehygiene against micro flora and/or in supplying pro-vitamin D and repository and excretory function forseveral pesticides and pollutants.

Removal of the uropygeal gland(uropygealectomy) from chicks deprived of uropygeallipids which causes significant increase in growthwhen commercial chick-feed was fed ad libitum tothe operated and sham-operated chickens (Haahtia etal, 1964). Zhang et al. (2010) studied the uropygealgland-secreted alkanols contribution to olfactory sexsignals in budgerigars. The behavioral data robustlysuggested that the 3 alkanols synergistically createda female attractant odor or male pheromone in thebudgerigar and that bird uropygeal glands had broaderimplications in sexual behavior than previously

known. France investigator Giraudeau (2010)established a new link between important avian self-maintenance behavior and aspects of maternal healthand reproduction. They suggested that higher yolkcarotenoid levels in eggs laid by preen-gland-restricted females may serve to boost health ofoffspring that would hatch in a comparativelymicrobe-rich environment. According to the author’sbest knowledge, there have been a very limited workdone associated with studying the serum biochemicalconstituents in young hens and old hens after removalof preen gland in relation to egg production.


The present research work was carried out forestimation of serum biochemical constituents inchicken (gallus domesticus) before and afterextirpation of the preen gland in relation to eggproduction. The experimental 100 White Leghornlayers were divided into two age group i.e. (A) YoungHen aged 32 wks. old and (B) Spent Hen aged 90wks. old, comprising 50 birds in each group. Theyoung hens were again subdivided into Group I -Start control group (10 birds), Group II - End control

58


group (20 birds) and Group III - preen glandextirpated group (20 birds). The spent hens were againsubdivided into Group IV - Start control group (10birds), Group V - End control group (20 birds) andGroup VI - preen gland extirpated group (20 birds).

The standard procedure of scientific feeding andmanagement were followed throughout theexperiment. Sixteen hours of light and eight hours ofdarkness was provided to the birds throughout theexperimental period of six weeks.

At beginning of the experiment, 10 hens fromeach Group I and Group IV were sacrificed and bloodserum samples were collected. Preen gland of 20 birdswere surgically removed from each of Group-III andGroup-VI, simultaneously, Group II and Group IVacts as end control groups. Then at the end of everyweeks, pulled blood serum samples were collectedfrom Group II, III, V and VI, for six consecutive weeksand stored at – 20o C. The blood samples about 2 – 3ml, were collected aseptically from the jugular veinin separate sterile glass test tubes withoutanticoagulant in morning hours between 9 am and 11am. Equal volume of blood serum were taken fromfive blood samples and pooled them to make sufficientvolume of “Pulled blood serum”. The blood serumconstituents viz. serum total cholesterol, triglycerides,calcium and phosphorous were analyzed using anauto-analyzer-STAT FAX 2000.


The study was conducted to quantify andcompare the circulating levels of some serumbiochemical constituents, before and after removal ofpreen gland in relation to egg production in youngand spent White Leghorn hens/layers.

1. Total Cholesterol :The mean ± SE values of serum total

cholesterol in White Leghorn layers in differentgroups are presented in Table No. 1 and itsanalysis of variance in Table No. 2.

In young hens, the serum total cholesterolconcentration was 117.20 ± 3.17, 127.23 ± 6.87and 138.16 ± 10.28 mg/dl in Group I, II and III,respectively. In spent hens, the serum totalcholesterol concentration was 133.60 ± 28.05,136.89 ± 31.27 and 138.57 ± 34.66 mg/dl in

Group IV, V and VI, respectively.Higher level of total cholesterol

concentration was recorded in preen glandextirpated groups in both young and spent hensthan in start and end control groups. Thesefindings corroborates with the finding of Hassan(2010) in Japanese quail. The small differencesin serum total cholesterol concentration werestatistically non-significant.

The results of total cholesterol concentrationcould not be compared as no reports could betraced. The level of serum total cholesterolconcentration in young and spent hens recordedin the present study was within the ranges asreported by Sturkie (1975). Sturkie (1975)reported that, the concentration of serumcholesterol range was 83 to 132 mg/dl in aves.Hassan (2010) reported that the plasma totalcholesterol concentration increased withadvancement in age up to 30 weeks of age whichwas 302 mg/dl in Japanese quail. They foundhighly significant positive phenotypic correlationof plasma total cholesterol with age and numberof eggs. Bhatti et al. (2002) estimated serumcholesterol during pre-laying and layingconditions in Desi, Fiyoumi, Nick chick andCross (RIR X Fiyoumi) as 108.69, 116.75,115.00 and 143.33 mg/dl respectively. Silva etal. (2007) in HYBRO-PG broilers reported that,the serum cholesterol level was 140.16 ± 20.34,128.9 ± 16.39 and 129.42 ± 20.36 mg/dl in 21days, 35 days and 42 days old broiler,respectively. Montalti et al. (2006) however,reported that the removal of preen gland did notaffect serum total cholesterol concentration inrock pigeons.

2. Triglycerides :The mean ± SE values of serum triglycerides

in White Leghorn layers in different groups arepresented in Table No. 3 and its analysis ofvariance in Table No. 4.

In young hens, the serum triglycerideconcentration was 1130.00 ± 32.80, 1187.57 ±86.90 and 1208 ± 62.03 mg/dl in Group I, II andIII, respectively. In spent hens, the serumtriglyceride concentration was 1004.00 ± 40.42,1028.43 ± 75.41 and 1036.70 ± 60.52, mg/dl in

59


Table No. 1: The mean ± SE values of serum total cholesterol in White Leghorn layers

Biochemical

constituent

(mg/dl)

Young hens Spent hens

Group I Group II Group III Group IV Group V Group VI

Total

cholesterol

117.20 ±

3.17

127.23 ±

6.87

138.16 ±

10.28

133.60 ±

28.05

136.89 ±

31.27

138.57 ±

34.66

Table No. 2: Analysis of variance of the data of serum total cholesterol

Sources of variation DF MSS F’ Value

Groups 5 489.033 0.805NS

Weeks 6 215.599 0.351

Error 30 607.071 -

NS = Non-significant.

Table No. 3: The mean ± SE values of serum triglycerides in White Leghorn Layers

Biochemical

constituent

(mg/dl)

Young hens Spent hens C. D.

Value

5% Group I Group

II

Group

III

Group

IV

Group

V

Group

VI

Triglycerides 1130.00

± 32.80

1187.57

±86.90ab

1208 ±

62.03a

1004.00

± 40.42

1028.43

± 75.41c

1036.70

±60.52bc

159.14

Note: Those means with at least one common superscript do not differ significantly.

60


Group IV, V and VI, respectively. The differencein triglyceride concentration in preen glandextirpated and start control group of young andspent hens was non-significant. The highesttriglycerides concentration was observed in preengland extirpated group when compared with theircontrols. It could be attributed to highermetabolic rate which was related to more eggproduction in both groups. The difference inserum concentration in preen gland extirpatedgroup was significantly higher than that in groupsfrom I to VI.

The serum triglycerides concentration in endcontrol and preen gland extirpated groups wassignificantly higher (p < 0.05) than in end controland preen gland extirpated groups in spent hens.

The results of serum triglyceridesconcentration could not be compared as noprevious reports could be traced. Sturkie (1975)reported that, the concentration of serumtriglycerides was 59.7 % wt. of lipid. In thepresent study, higher serum triglyceridesconcentration was recorded in preen glandextirpated groups in both young and spent henscorroborates with the findings of Hassan (2010)in Japanese quails who reported that the plasmatriglycerides concentration increased withadvancement in age up to 30 weeks of age whichwas 652.00 mg/dl in 30 weeks of age. They foundhighly significant positive phenotypic correlationof plasma triglycerides with age and number ofeggs. Silva et al. (2007) in HYBRO-PG broilers

reported that, the serum triglycerides level was130.8 ± 28.03, 97.11 ± 25.16 and 132.52 ± 33.52mg/dl in 21 days, 35 days and 42 days old broiler,respectively. Montalti et al. (2006) reported thatthe removal of preen gland did not affect serumtriglycerides concentration in rock pigeons.

Thus, from the result, it can be concludedthat extirpation of the preen gland in both youngand spent hens, increased the serum triglyceridesconcentration.

3. Calcium :The mean ± SE values of serum calcium in

White Leghorn layers in different groups arepresented in Table No. 5 and its analysis ofvariance in Table No. 6.

In young hens, the mean serum calciumconcentration was 19.00 ± 1.04, 19.97 ± 0.28and 21.70 ± 1.45 mg/dl in Groups I, II and III,respectively. In spent hens, the serum calciumconcentration was 20.30 ± 0.44, 20.57 ± 0.77and 20.90 ± 0.52 mg/dl in Group IV, V and VI,respectively. The difference in calciumconcentration in preen gland extirpated (GroupIII) and end control (Group II) group wasstatistically significant. The calciumconcentrations in start control group of youngand spent hens was apparently lower than in endcontrol and preen gland extirpated groups.

The results of serum calcium concentrationcould not be compared as no previous reportscould be traced. The level of serum calcium

Table No. 4: Analysis of variance of the data in serum triglycerides


Groups 5 54002.809 2.542*

Weeks 6 8815.683 0.416

Error 30 21258.526 -

* = Significant at 5% level.

61


concentration in young and spent hens recordedin the present study was within the ranges asreported by Etches (1987) in laying hens. Hestated that, the plasma concentrations of Ca2+ aremaintained between 20.00 and 25.00 mg/100mlthroughout the ovulatory cycle and period ofshell formation. Similar was the finding ofRoepke and Hughes (1934) who reported that,the serum calcium level was 20.00 mg/dl inlaying hens. Celebi et al. (2005) studied theinfluence of dietary phosphorus level in plasmacalcium in 70 weeks old 192 White Lohmanlaying hens. They found 9.7 mg/dl of serumcalcium. However, Burkholder (1970) in red-

winged birds, stated that, the birds sampled fromthe field during the process of eggshellcalcification had an average serum calcium levelof 31.84 mg% with a range of 14.85 to 61.73mg%. Bhatti et al. (2002) reported mean serumcalcium levels were 4.887, 5.17, 5.96 and 6.56mg/dl in Desi, Fiyoumi, Cross (RIR X Fiyoumi)and Nick chick, respectively in egg layingcondition. Silva et al. (2007) in HYBRO-PGbroilers reported that, the serum calcium levelwas 9.54 ± 1.65, 11.06 ± 2.07 and 8.85 ± 1.35mg/dl in 21 days, 35 days and 42 days old broiler,respectively.

In the present study higher serum calcium

Table No. 5: The mean ± SE values of serum calcium in White Leghorn layers

Biochemical

constituent

(mg/dl)

Young hens Spent hens C. D.

Value

5% Group

I

Group

II

Group

III

Group

IV Group V

Group

VI

Calcium

19.40 ±

1.04 19.97 ±

0.28b

21.70

±1.45a

20.30 ±

0.44

20.57 ±

0.77abc

20.90

±0.52ab

1.25

Note: Those means with at least one common superscript do not differ significantly.

Table No. 6: Analysis of variance of the data of serum calcium


Groups 5 4.398 3.337*

Weeks 6 1.746 1.328

Error 30 1.313 -

*= Significant at 5% level.

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concentration was recorded in preen glandextirpated groups in both young and spent henswhich corroborates the very recent findings ofHassan (2010) in Japanese quails who reportedthat the plasma calcium concentration increasedwith advancement in age up to 30 weeks of age.The plasma calcium concentration was 31.99 mg/dl in 30 weeks of Japanese quail. They foundhighly significant positive phenotypic correlationof plasma calcium with age and number of eggs.However, Montalti et al. (2006) reported that theremoval of preen gland did not affect serumcalcium concentration in rock pigeons.

The higher serum calcium concentration inyoung than in spent hens could be attributed tohigher metabolic rate in young hens which wasrelated to more egg production. Sturkie (1975)reported that, the concentration of serum calciumrises from 10 mg% to 16-30 mg% during the 10days before a pullet starts to lay. A hen that lays250 eggs per year secretes a quantity of calcium,in the form of shells, corresponding to about 20times the calcium contents the calcium contentsof her entire body. During the last 15 hrs. of shellformation, calcium moves across the shell glandof the hen at the rate of 100 – 150 mg/hr.

Thus, from the results, it could be concludedthat extirpation of the preen gland in both youngand spent hens, increased the serum calciumconcentration.

4. Phosphorous :The mean ± S.E. values of serum

phosphorous in White Leghorn layers in differentgroups are presented in Table No. 7 and itsanalysis of variance in Table No. 8.

In young hens, the serum phosphorousconcentration was 6.20 ± 0.60, 7.35 ± 0.49, and7.43 ± 0.43 mg/dl in Group I, II and III,respectively. In spent hens, the serumphosphorous concentration was 7.50 ± 0.60, 8.53± 0.68 and 8.71 ± 0.80 mg/dl in Group IV, V andVI, respectively. In young and spent hens, theserum phosphorous concentration was higher inpreen gland extirpated group than in start andend control groups. The small difference in theserum phosphorous concentration, however, didnot approach statistical significance.

The serum phosphorous concentration wassignificantly higher (p < 0.05) in Group VI thanGroup II. In remaining groups, the concentrationdid not significantly differ and remained almostthe same.

The results of serum phosphorousconcentration could not be compared as noprevious reports could be traced. The level ofserum phosphorous concentration in young andspent hens recorded in the present study arewithin the ranges as reported by Roepke andHughes (1934) who stated that, the serumphosphorous level was 9.70 mg/dl in laying hens.Celebi et al. (2005) studied the influence ofdietary phosphorus level in 70 weeks old 192White Lohman laying and found 4.60 mg/dl ofserum phosphorus. Egg shell quality measuredby specific gravity was inversely related todietary phosphorus levels above 0.50 %. Bhattiet al. (2002) found serum phosphorous duringpre-laying and laying conditions in Desi,Fiyoumi, Nick chick and Cross (RIR X Fiyoumi)as 6.1, 8.05, 6.83 and 6.81 mg/dl respectively.Silva et al. (2007) in HYBRO-PG broilersreported that, the serum phosphorus level was6.23 ± 0.75, 6.81 ± 0.98 and 6.12 ± 0.83 mg/dlin 21 days, 35 days and 42 days old broiler,respectively.

In the present study, the serum phosphorousconcentrations increased in preen glandextirpated groups in both young and spent henswhich is in accordance with Hassan (2010) inJapanese quails who reported that the plasmaphosphorous concentration increased withadvancement in age up to 30 weeks of age whichwas 8.10 mg/dl in Japanese quail. They foundhighly significant positive phenotypic correlationof plasma phosphorous with age and number ofeggs. In present study, the increase inphosphorous concentration in spent hen mightbe due to old age as described by Hassan (2010).Montalti et al. (2006) however, reported that theremoval of preen gland did not affect serumphosphorous concentration in rock pigeons.

The higher serum calcium concentration inyoung than in spent hens could be attributed tohigher metabolic rate in young hens which wasrelated to more egg production.

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Thus, from the present results, it could beconcluded that extirpation of the preen gland inboth young and spent hens increased the serumphosphorous concentration.

The results of the present study indicated thatthe serum total cholesterol, triglycerides, calciumand phosphorous were increased as the ageadvances.

SUMMARY AND CONCLUSIONS

Higher level of serum total cholesterolconcentration was recorded in preen gland extirpatedgroups in both young and spent hens than theircontrols, the small differences in serum totalcholesterol concentration were statistically non-significant. The serum triglycerides concentration inend control and preen gland extirpated groups wassignificantly higher (p < 0.05) than in end control andpreen gland extirpated groups in spent hens. Inremaining groups, the concentration did notsignificantly differ and remained almost the same.

From the present results following conclusionswere drawn :

The serum biochemical constituents like calcium,phosphorous, triglycerides and total cholesterol wereincreased as the age advanced. Further, theconcentration of these biochemical constituents wasfound higher in preen gland extirpated groups than intheir respective controls.

REFERENCES

Bhatti B. M., Talat, T. and Sardar, R. (2002).Estimation of serum alkaline phosphatase, cholesterol,calcium and phosphorous during pre-laying and layingconditions in different strains of chickens. PakistanVet. J. 22 (2): 94 – 96.

Burkholder, T. (1970). Serum calcium levels of theRed-winged black bird (Agelaius phoeniceus). Site:http://digitalcommons.unl.edu/icwdmbirdcontrol/214.

Celebi S., Bolukbasi, S. C. and Utlu, N (2005). Theinfluence of dietary phosphorus level on plasma

calcium and phosphorus, egg shell calcium andphosphorus. Int. J. Poultry Sci., 7(4) : 497 – 499.

Etches, R. J. (1987). Calcium Logistics in the LayingHen. J. Nutr. 117: 619.

Giraudeau M., Czirják, G. A., Duval, C., Bretagnolle,V., and Eraud, C. (2010). Effect of Restricted Preen-Gland Access on Maternal Self Maintenance andReproductive Investment in Mallards.

Haahtia E., Lagerspetza, K., Nikkaria, T and Falesa,H. M. (1964). Lipids of the uropygeal gland of birds.Comparative Biochemistry and Physiology, 12(3):435 – 436.

Hassan, H. A. (2010). Variations in egg performanceand plasma constituents at different ages of femaleJapanese quail. Egypt. Poult. Sci. 30(2) : 565 – 581.

Montalti, D., Gutiérrez, A. M., Reboredo, G. R. andSalibián, A. (2006). Removal of the uropygeal glanddoes not affect serum lipids, cholesterol and calciumlevels in the rock pigeon Columba livia. Acta BiolHung. 57(3): 295 – 300.

Roepke, R. R. and Hughes, J. S. (1934). Phosphoruspartition in the blood serum of laying hens*www.jbc.org

Silva P. R.L., Freitas Neto, O. C., Laurentiz, A. C.,Junqueira, O. M. and Fagliari, J. J. (2007). Bloodserum components and serum protein test of Hybro-PG broilers of different ages. Rev. Bras. Cienc. Avic.9 (4).Site - http://dx.doi.org/10.1590/S1516-635X2007000400004.

Sturkie, P. D. (1975). Avian Physiology, Third edition,Pp – 302 – 371.

Zhang, J. W. Wei, J. H., Zhang and Yang, W. (2010).Uropygial Gland-Secreted Alkanols Contribute toOlfactory Sex Signals in Budgerigars. OxfordJournals, Life Sciences & Medicine, Chemical Senses,35 (5): 375-382.

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Constraints of Dhangar sheep pastoralists’ community in Western Maharashtra state.

Ajit Pokharkar, S.T. Hande*, M.N. Sawant, D.N. Kale and T.R. NikamDepartment of Veterinary and Animal Husbandry Extension, Bombay Veterinary College, Parel, Mumbai.

*Department of Livestock Production and Management, Bombay Veterinary College, Parel, Mumbai.

ABSTRACT

The present study was carried out to analyse the constraints faced by the Dhangar sheeppastoralists community in Western Maharashtra region. The study was conducted in twodistrict of western Maharashtra by personally interviewing 120 Dhangar tribe rearing sheep.Here majority of respondents having scarcity of fodder and water, incidences of differentdiseases, decreasing grazing land, non availability of credit/loan, lack of technical guidance,lack of proper training, lack of veterinary aid & high cost of medicines, poisoning throughherbicide, lack of superior male animal, non availability of labour, lack of transport facilitiessocial norms and taboos, lack of knowledge and these were found to be the major constraintsfaced by the respondents involved in sheep husbandry.

INTRODUCTION

In Indian context, pastoralists can be defined as“member of caste or ethnic group with a strongtraditional association with livestock-keeping, wherea substantial proportion of the group derive over 50per cent of household consumption from livestockproducts or their sale, and around 90 per cent of animalconsumption is from natural pasture or browse(Sharma et al. 2003). The state of Maharashtra alonehas 11.23 crore human population which is 9.29 percent of India. Dhangar is the traditionally semi-nomadic pastoral society primarily located in the stateof Maharashtra. Dhangars maintain variety oflivestock. In this context an attempt is made here tofind out the long term persistent obstacles faced byDhangar pastoralist in running their sheep rearingoccupation .The Schedule was developed whichincludes physical, technical, economical andoperational constraints. Constraints were rankedaccording to Garrett ranking method USAID (2010).

MATERIAL AND METHODS

Locale of the study : The present study wascarried out in the Maharashtra state. The state waspurposively selected due of the following reasons:

● Dhangar is the traditionally nomadic pastoralsociety primarily located in the state ofMaharashtra.

● Dhangar caste-cluster comprises of 23endogamous castes which are spread in all districtof Maharashtra. Total population of Dhangarnomadic tribe in Maharashtra is nine million.

● Maharashtra has 32.75 lakh sheep population andit ranks sixth in sheep population (Livestockcensus 2007, Department of animal husbandryMaharashtra).

● Maharashtra ranks third in Meat production &eighth in wool production amongst all states ofIndia (Source: Basic Animal HusbandryStatistics, 2006).

● Selection of districtMaharashtra state comprises 35 districts out of

these eight districts namely Pune, Satara, Sangli,Solapur, Kolhapur,Latur, Parbhani and Nanded arehome land of Dhanger nomadic tribe. Two districtsnamely Sangli and Satara district was selectedpurposely for the present study because more numberof Dhangar population inhabited in this district. Thesetribe used to keep sheep (Decani, Madgyal and Non-Descript), goat (Osmanabadi, Sangmneri and Non-Descript), along with these livestock they keep fewcattle and buffalo.

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Selection of the respondents :From each district two tehsils were selected

randomly and two villages from each tehsil wereselected randomly. In this way total eight villages wereselected from both the districts. From each village 15respondents were selected randomly which belongedto Dhangar tribe and having minimum one type oflivestock. They were interviewed with the help ofinterview schedule keeping in view the objectives ofthe study. Thus total respondents for the study were120 from the both districts of Maharashtra state.

Tools and techniques for data collection :

The basic instrument used for the study was theinterview schedule (Freeman et al. 2007). Thequestions were related to different constraints facedby the Dhangar Sheep pastoralists community .

Result and Discussion :

The Dhangar community faces much constraintin their day to day life some of major are listed intable 1.

From the table it is depicted that “scarcity offodder and clean drinking water” was ranked first (94

%). It was the most important perceived constraintsin livestock rearing. Since, it is not possible to keepanimals without feed and water, therefore respondentstrouble to maintain more no of animals. The secondimportant perceived constraint was incidences ofdifferent diseases (92.66 %). Prevention of diseasesand to maintain good health of animals requires manystrategies.

Lack of grazing land were perceived as third(91.33%) important constraint in livestock rearing.Dhangars totally depended on others grazing landfor their livestock. Due to decreasing grazing land orunavailability of grazing land the people avoid to keepmore number of animals as consequences decreasetheir income.

It can be concluded from the table that nonavailability of credit is the major constraint facedby the respondents, since majority of them were lesseducated & illiterates. Majority of the respondents aretaking a livestock health facility from privateveterinarian. So they did not get much exposure tothe government veterinary officer to locate theavailability of credit from various sources and variousschemes launched by government for sheep farmers(Krishna et al. 2004). This constraint of veterinaryaid should be tackled immediately to minimize the

Table 1. Constraints faced by Dhangar in sheep husbandry.

Sr. No Constraints Frequency(n) Percentage(%) rank1. Scarcity of fodder & water 113 94 I

2. Incidences of different diseases 110 92.66 II

3. Lack of grazing land 109 91.33 III

4. Non availability of credit and loan 102 85.33 IV

5. Lack of technical guidance. 95 79.33 V

6. Lack of proper training. 89 74.66 VI

7. Lack of veterinary aid &High cost of medicine. 79 66 VII

8. Poisoning through herbicide during grazing. 77 64.66 VIII

9. Lack of superior quality male animal 74 62.66 IX

10. Non availability of labor. 72 60.66 X

11. Lack of transport facilities. 71 59.33 XI

12. Social norms and taboos. 50 42 XII

13. Lack of knowledge. 46 38.66 XIII

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animal health risk. Some time in governmentveterinary dispensary, there is no availability ofimportant vaccines, artificial insemination facility onrequired time which leads to heavy loss in productionas well as health measures. So veterinary aid is alsoone of the major constraints faced by the respondents.

The constraints of Poisoning through herbicideduring grazing were ranked as eight (64.66 %) by therespondents. According to the respondents lack ofsuperior quality male animal for breeding purpose wasthe ninth important constraint (62.66%), which isimportant for increase the size of flock or number ofgood quality animals.

Non availability of labour is also one of theimportant constraints faced by respondents. As nearbycity has vigorously developed as an industrial area.People are getting a lot of opportunities, jobs withattractive payments in the industrial companies. Hencethey preferred these jobs instead of sheep husbandry.This leads to one of the important constraint for sheephusbandry practices.

The next difficulty expressed by the Dhangar wasthe lack of transport facilities. The sample villageswere remotely connected with bus facilities, but it isnot available on timely which resulted in difficulty intransporting their produce in time, causingdeterioration in quality and economic loss to therespondents.

Social norms and taboos were considered as thenext constraint faced by the Dhangar involved in sheephusbandry practices. They opined that age oldtraditions, social attitudes and scientific prejudices,myths and beliefs were the major constraints in theirparticipation in agricultural activities.

Conclusion and Implications of the study.

Scarcity of fodder and water, incidences ofdifferent diseases, decreasing grazing land, nonavailability of credit/loan, lack of technical guidance,lack of proper training, lack of veterinary aid & highcost of medicines, poisoning through herbicide, lackof superior male animal, non availability of labour,lack of transport facilities social norms and taboos &

lack of knowledge were found to be the majorconstraints faced by the respondents involved insheep husbandry.

The remedies and recommendations to overcomethe constraints faced by the respondents involved insheep rearing may include proper training should begiven to the respondents to improve knowledge &skill regarding sheep husbandry practices.Government should provide more funds to ruralfinancing institutions, voluntary agencies, NGOs forproviding loans and services to rural women.Government should also take efforts to providecrossbred animals, feed and medicines at reasonableprices to the Dhangar.

Scientists, experts should organize a campaignwith actual demonstration regarding the scientificpreparation & storage of feeds & fodder like haymaking, silage making. It will help in avoiding theproblem of scarcity of green fodder. Technologieswhich are labour saving, income generating andproductivity increasing should be popularized. Alsorain water harvesting programmes may be encouragedin the study area to solve the problem of water scarcity.More emphasis should be given on mobile veterinaryclinic to avoid inconvenient veterinary service.

REFERENCES

Freeman, A., Kaitibie, S., Moyo, S. and Perry B.(2007). Livestock, livelihoods and vulnerability inLesotho, Malawi and Zambia. ILRI and FAO STAT(2005)

Krishna, A., Kristjanson P., Radeny, M. and Nindo,W., (2004). Importance of livestock, Journal of HumanDevelopment 5: 211-226.

USAID, (2010). Dry land livestock developmentprogramme. http://kenya.usaid.gov/programs/economic-growth/412

Sharma, V., Kochler-Rollefson and Morton, J. (2003).Pastoralism in India: A Scoping Study, IIM,Ahemdabad, India. Page No.22-36.

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Effect of Supplemental Chromium on Serum Cortisol and Growth Weight in Sirohi Goat Kids

S.V. Sangekar*, S. D. Ingole, S. V. Bharucha and A. S. NagvekarDepartment of Veterinary Physiology, Bombay Veterinary College, Parel, Mumbai.*Department of Biochemistry, Institute of Science, Mumbai University, Mumbai.

ABSTRACT

Twelve apparently healthy Sirohi kids from Instructional Livestock Farm Complex,Bombay Veterinary College, Unit No.3, Aarey, Goregaon, Mumbai were utilized for the presentstudy. The kids were divided into 2 groups of 6 kids aged between 2-4 months each as control(without 4% chromium) and treatment (with 4% chromium) at the dose rate of 7.5mg/goat/day. On comparing cortisol concentration in goat kids supplemented without 4% chromiumand with 4% chromium during growth showed that the levels decreased significantly (P<0.05) on the day 7 after supplementation of 4% Cr and further it declined on the day 28,whereas there was no change in the levels in the kids supplemented without 4% Cr.The weightgain in goat kids of both the groups did not show significanr difference, but the kidssupplemented with 4% chromium showed numerical increase in weight throughout, where asthe weight remained stable in the kids supplemented without 4% chromium from day 14 today 28.

Keywords: Goats, Chromium, Cortisol, Growth

INTRODUCTION

A majority of studies on heat stress in livestockhave focused mainly on temperature and relativehumidity which may exert negative effects on farmanimal welfare, performance and health. Stress whichis defined as “abnormal pressure, strain, constrainingforce or influence” can occur in biological systems invarious forms from very mild dietary stresses to severephysical trauma or injury. Chronic stresses may alsoalter nutrient requirements. If dietary intake issuboptimal, a small stress-induced alteration may havesignificant effects on the signs and symptoms ofdeficiency.Thus there is an urgent need for detailedassessment for identifying appropriate options that canhelp livestock to adapt to such climate and overcomestress. One metal that is suboptimal in the diet andstrongly influenced by stress in humans and farmanimals is chromium (Cr).

Supplementation of Cr in livestock diets mayimprove animal metabolism and enhance productionperformance and the quality of animal products

(Spears, 1999). Trivalent chromium is essential fornormal carbohydrate, lipid and protein metabolism.The demand for Cr has been growing as a result offactors commonly referred to as stressors, especiallyduring different forms of nutritional, metabolic andphysical strain. It has been confirmed from studiesthat Cr is associated with the metabolism duringincreased physiological, pathological and nutritionalstress. Cr demand in humans and animals increasesduring periods of higher stress – e.g. fatigue, trauma,gestation and different forms of nutritional (high-carbohydrate diet), metabolic, physical, and emotionalstress as well as environmental effects (Anderson,1994). It has been shown Cr reduces the negativeeffects of environmental stress (Sahinet al., 2001;Mowat, 1994; Lien et al., 1999).

A number of authors confirm decreasedsensitivity to stress in Cr supplemented animalsthrough a reduced concentration of cortisol in blood(Chang and Mowat, 1992; Moonsie-Shageer andMowat, 1993; Mowat et al., 1993; Pechovaet al.,2002).The effect of Cr supplementation on the

68


intensity of growth has been studied especially in pigsand cattle. In cattle, a positive effect of Crsupplementation on weight gain has been recordedby Chang and Mowat (1992), Moonsie-Shageer andMowat (1993) and Kegleyet al. (1997). Despite thefact that the results have been ambiguous, most authorsagree that Cr supplementation during periods ofincreased stress has a positive effect on weight gain.

The data show that a positive effect of chromiumsupplementation in feeding rations has beendemonstrated in only 40–50% of experiments,depending on the animal category and the effect ofchromium supplementation monitored. Chromium hasthe benefit of high safety, verified by the fact that theexperiments revealed no negative effect of trivalentchromium on the animal’s health status.

Since Cr supplementation has shown intenseeffect in animals, but less studied in goats duringgrowth, this research is taken to study theeffects ofdietary Cr and weight gain in goat kids.

MATERIALS AND METHOD

Twelve apparently healthy Sirohi kids fromInstructional Livestock Farm Complex, BombayVeterinary College, Unit No.3, Aarey, Goregaon,Mumbai were utilized for the present study. The kidswere divided into 2 groups of 6 kids aged between 2-4 months each as control (without 4% chromium) andtreatment (with 4% chromium).

The goats were maintained under uniformstandard conditions of feeding and management. Theanimals were kept in collective pens with asbestosroof, under natural daylight and temperatureconditions. The goat kids were let loose for grazingand exercise from 7:00 am to 11:30 am. Then at12:00pm, they were each provided with goat feed forkids, procured from Godrej Agrovet Ltd, Mumbai,along with green roughages. Clean drinking water wasavailable. Both the groups were fed with the abovesaid feed, 3-4% of body weight twice a day, while thetreatment group was supplemented along with 4% Crat the dose rate of 7.5mg/goat/day. During the periodof experiment the average Temperature HumidityIndex (THI) was calculated, using the dry bulb andwet bulb temperature of the pens which was 30Ú and28ÚC respectively. The THI was 82.36%.

Blood samples (8-10ml) were collected

aseptically from the jugular vein in the separate glassvacutainers without anticoagulant. Samples will becollected in the morning hours between 8.00 a.m. and10 a.m. Clear serum will be separated bycentrifugation and will be stored at -20ÚC. Thequantitation of cortisol was done using RIA kitsprocured from Immunotech, A Beckman CoulterCompany, Immunotech SAS – 130 av. De Lattre deTassigny – B.P.177 – 13276, Marseille Cedex 9,France.

RESULT AND DISCUSSION

CortisolThe results of serum cortisol are presented in

table 1 and in figure 1 for Sirohi goat kidssupplemented without 4% Cr and supplemented with4% Cr.

The difference in serum cortisol concentrationduring the experiment in kids supplemented without4% Cr were statistically non-significant with minorfluctuation. In contrast Depew et al. (1998) reporteddecrease in cortisol concentration in young dairycalves during growth.

The difference in the serum cortisolconcentration during the experiment in kidssupplemented with 4% Cr were statistically significant(P < 0.05).

The results of the present study are in agreementwith Haldaret al. (2007) in Black Bengal goats. Theyreported a significant decrease in the serum cortisolin those supplemented with Cr, compared with thegroup supplemented without Cr. The role ofsupplemental Cr in reducing cortisol level inruminants is known (Chang and Mowat, 1992;Moonsie-Shageer and Mowat 1993; Mowatet al. 1993and Haldaret al. 2006). Most of the findings showingreduction in serum cortisol concentration due to Crsupplementation are involved in stress factors liketransportation or vaccination. In the present study thestress may be due to environment temperature wherein the Temperature Humidity Index (THI) was above70, which indicates little discomfort to the animals.Beside cortisol having its relation to stress, it also hasits role as immunosuppressant and anti-inflammatory(Borgs and Mallard, 1998). This role of cortisol on itsreduction after Cr supplementation may help in

69


enhancing the performance of the goat kids.Similarly results were also reported by Chang

and Mowat (1992) and Moonsie-Shageer and Mowat(1993) and Depew et al. (1998) in growing feedercalves supplemented with Cr which decreased cortisollevels. According to them stress increases serum

glucocorticoid levels. Glucocorticoids promotehyperglycemia by stimulating gluconeogenesisreducing glucose utilization (Bennetet al 1998;Brockman, 1986). Moreover, cortisol also inhibits theproduction and action of cytokines and antibodieslymphocyte function and leukocyte population (Roth

Table 1. Serum Cortisol Concentration (nmol/l) during growth in Sirohi goat kids supplemented without 4% Cr and with 4% Cr.

DAYS CONTROL (Without 4% chromium)

TREATMENT (With 4% chromium)

0 3.73 ± 0.23 2.85a ± 0.51

7 3.48 ± 0.27 1.42b ± 0.31

14 3.31 ± 0.31 0.37c ± 0.18

21 3.50 ± 0.31 0.20c ± 0.02

28 3.72 ± 0.29 0.17c ± 0.02

3.73 3.48

3.31 3.5

3.72

2.85

1.42

0.37 0.2 0.17

0

0.5

1

1.5

2

2.5

3

3.5

4

0 7 14 21 28

Cor

tisol

Con

cent

ratio

n (n

mol

/l)

Days

controltreatment

Fig. 1 : Serum cortisol concentration (nmol/l) during growth in Sirohi goat kids supplemented with and without 4% Cr.

70


and Kaeberle, 1982; Muncket al., 1984). Thusdecrease in cortisol will enhance overall production.

On comparing the cortisol concentration in boththe group, it was observed that the levels decreasedsignificantly in the goat kids supplemented with 4%Cr, whereas there was no change in the levels in thekids supplemented without 4% Cr.

Body WeightThe results of the body weight are presented in

the table 2 and in figure 2 for Sirohi goatssupplemented without 4% Cr and supplemented with4% Cr.

The difference in average body weight gainduring the experiment in kids supplemented without4% Cr were statistically non-significant butnumerically there was an increase in the body weighton day 7 and 14 and further it remained almostconstant up to day 28 with minor fluctuations.

The difference in average body weight gainduring the experiment in kids supplemented with 4%Cr were statistically non-significant but numericallythere was an increase in the body weight throughouton day 7, 14, 21 and 28, respectively from 5.51kg to8.05 kg.

On comparing both the group, though there wasnon-significant increase in body weight, but the gainin kids supplemented with 4% Cr was morethroughout, while in kids supplemented without 4%Cr remained almost constant after 14 days.

The results of the present study are in agreementwith Haldaret al. (2007) in Black Bengal goats. Theyalso reported non-significant increase in body weightin Cr supplemented kids and in control group. But oncomparing the control and treatment group, theyreported similar increase in body weight, which is incontrast to present study, where there was numericalincrease in the weight gain till day 28 in treatment(supplemented with Cr) group, while the control(supplemented without Cr) group showed increase inweight gain up to day 14 and then remained almostconstant.

Further the findings of the present study tell usthat the average body weight gain throughout in Crsupplemented kids corroborates with reports instressed feeder calves (Moonsie-Shageer and Mowat,1993) and growing feeder calves (Chang and Mowat,1992). According to them this increased weight gainscould be due to Cr action via cortisol. The decreasedcortisol may cause improved immune response sinceglucocorticoids are known to inhibit natural killeractivity, lymphocyte activating factors and T-cellgrowth factor production.

Thus it can be concluded that on supplementing4% Cr to the goat kids, the levels of cortisol decreaseswhich is an indicator of decrease in stress. Thisdecrease helps in increase in the feed intake and bodyweight of the animal, thus improving the overallperformance.

Table 2. Average Weight Gain (kg) during growth in Sirohi goat kids supplemented without 4% Cr and with 4% Cr.

DAYS CONTROL

(Without 4% chromium) TREATMENT

(With 4% chromium)

0 7.96 ± 0.46 5.51 ± 0.86

7 8.72 ± 0.58 6.15 ± 0.88

14 9.11 ± 0.81 6.64 ± 0.93

21 9.70 ± 0.96 7.35 ± 1.15

28 9.52 ± 0.96 8.05 ± 1.34

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Fig. 2 : Average weight gain (kg) during growth in Sirohi goat kids supplemented with and without 4% Cr.

REFERENCES

Anderson, R. A. (1994). Stress effects on chromiumnutrition of humans and farm animals. In: Proceedingsof Alltech’s 10th Annual Symposium, Biotechnologyin the Feed Industry, Lyons P., Jacques K. A. (eds.),Nottingham University Press, UK, 267–274.

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Borgs, P. and Mallard B. A. (1998). Immune-endocrine interaction in agricultural species:chromium and its effect on health and performance.Domestic Animal Endocrinology, 15: 431-438.

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Chang, X. and D. N. Mowat (1992). Supplementalchromium for stressed and growing feeder calves. J.Anim Sci. 70:559-565.

Depew, C. L., L. D. Bunting, J. M. Fernandez, D. L.Thompson, Jr. and R. W. Adkinson (1998).Performance and metabolic responses of young dairycalves fed diets supplemented with chromiumtripicolinate. J Dairy Sci. 81:2916–2923.

Haldar S., T.K. Ghosh, M.C. Pakhira and De K (2006)Effects of incremental dietary chromium (Cr3+) ongrowth, hormone concentrations and glucoseclearance in growing goats (Capra hircus). Journal ofAgricultural Science, Cambridge. 144: 269-280.

Haldar, S., S. Samanta, R. Banarjee, B. Sharma andT. K. Ghosh (2007). Glucose tolerance and serumconcentrations of hormones and metabolites in goats(Capra hircus) fed diets supplemented with inorganicand organic chromium salts. Animal. 1: 347–356

Kegley, E.B., J. W. Spears and T. T. Brown (1997).Effect of shipping and chromium supplementation onperformance, immune response, and disease resistanceof steers. Journal of Animal Science. 75: 1956–1964.

Lien, T. F., Y. M. Horng and K. H Yang (1999).Performance, serum characteristics, carcass traits andlipid metabolism of broilers as affected by supplement

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of chromium picolinate. Br. Poult. Sci. 40: 357–363.

Moonsie-Shageer, S and D. N.Mowat (1993). Effectof level of supplemental chromium on performance,serum constituents, and immune status of stressedfeeder calves. J Anim Sci.71:232-238.

Mowat D. N. (1994). Organic chromium: a newnutrient for stressed animals. In: Proceedings ofAlltech’s 10th Annual Symposium, Biotechnology inthe Feed Industry, Lyons P., Jacques K. A. (eds.),Nottingham University Press, UK.275–282.

Mowat D.N., X. Chang and W. Z. Yang (1993).Chelated chromium for stressed feeder calves.Canadian Journal of Animal Science. 73: 49–55.

Munck, A., P. M. Guyre and N. J.Holbrook. (1984).Physiological functions of glucocorticoids in stressand their relation to pharmacological actions.Endocrinol. Rev. 525.

Pechova A., L. Pavlata andJ. Illek (2002). Metaboliceffects of chromium administration to dairy cows inthe period of stress. Czech Journal of Animal Science.47: 1–7.

Roth, J. A and M. L. Kaeberle (1982). The effect ofglucocorticoids on the bovine immune system. J. Am.Ved. Med. Assoc. 180:230.

Sahin, K. O. Kucuk, N. Sahin, and O. Ozbey (2001).Effects of dietary chromium picolinatesupplementation on egg production, egg quality andserum concentrations of insulin, corticosterone andsome metabolites of Japanese quails. Nutr. Res. 21:1315-1322.

Spears, J. (1999). Reevaluation of the metabolicessentiality of the minerals.Asian Australasian Journalof Animal Sciences. 12: 1002-1008.

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Micrometrical study on ventricle of heart in sheep (Ovis aries)

Kolekar V. P, Patil A. D, Dhande P. L, Shankhapal V. D, Aware N. N. and Jadhav P. M.Department of Veterinary Anatomy and Histology


ABSTRACT

The present micrometrical study was conducted on the ventricle of heart of six male andsix female apparently healthy sheep (Ovis aries). The average values of upper, middle andlower thickness of myocardium and pericardium were statistically highly significant andother all parameter were nonsignificant. The average values of micrometrical parameters ofventricle of heart were higher in male sheep than female sheep.

Keywords: Micrometry, Heart, Ventricle, Sheep.

INTRODUCTION

The heart pumps the blood continuously throughthe blood vessels by its rhythmic contraction. Theheart of sheep is principally composed of cardiacmuscle and connective tissue. The ventricles form thebulk of the heart. The knowledge of micrometricalstudy of sheep heart will be useful for understandingphysiological function as well as for diagnosticpurpose and post mortem examination (Getty, 1975).The present study is directed towards anatomicalinvestigation of heart of indigenous breeds of sheepas there is dearth of literature on micrometrical studyon ventricle of heart of the indigenous breeds of sheepin country in general and Maharashtra State inparticular.


Micrometrical observations were carried out onfollowing parameters of the right and left ventricle ofthe heart in sheep as per method of Culling (1969) bycalculating the average of 3 to 4 fields from eachprepared slides.

1. Thickness of endocardium (ìm)2. Thickness of myocardium (ìm)3. Thickness of pericardium (ìm)4. Distance between subsequent ‘z’ band (ìm)Statistical analysis was carried out as per the

method of Snedecor and Cochran (1994) for variousmicrometrical parameters of the right and left ventricleof sheep.

RESULTS AND DISCUSSIONThe results of micrometrical study of different

components of ventricle of heart are presented in tableno.1.

Thickness of endocardium (μm) :

The results revealed that the average thicknessof endocardium of the male heart in the region of upperright ventricle 32.61 ± 3.97 μm, similarly the averagethickness of endocardium of the female heart in theregion of upper right ventricle was 28.98 ± 3.62 μm .The average thickness of endocardium of the maleheart in the region of upper left ventricle was 28.98 ±3.62 μm whereas the average thickness ofendocardium of the female heart in the region of upperleft ventricle was 39.87 ± 3.64 μm. The averagethickness of endocardium of the male heart in theregion of middle right was of 36.25 ± 4.60 μm,similarly The average the thickness of endocardiumof the female heart in the region of middle rightventricle was 32.61 ± 3.97 μm . The average thicknessof endocardium of the male heart in the region ofmiddle left ventricle was 43.52± 3.99 μm .Theaverage thickness of endocardium of the female heart

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in the region of middle left ventricle was 43.52± 3.99μm . The average thickness of endocardium of themale heart in the region of lower right ventricle was34.44 ± 5.20 μm. Similarly the average thickness ofendocardium of the female heart in the region of lowerright ventricle was 32.61 ± 3.97 μm . The averagethickness of endocardium of the male heart in theregion of lower left ventricle was 43.52 ± 3.99 μm,whereas the average thickness of endocardium of thefemale heart in the region of lower left ventricle was38.06 ± 4.67 μm. These observations were statisticallynon significant between male and female sheep. Theseobservations were in accordance with the observationsreported by Masuda et al. (1989) in Canines.

Thickness of myocardium (μm) :

The average thickness of myocardium of themale heart in the region of upper right ventricle was5416.88 ± 86.13 μm, whereas the average thicknessof myocardium of the female heart in the region ofupper right ventricle was 5036.43 ± 114.58 μm. Theaverage thickness of myocardium of the male heartin the region of upper left ventricle was 6521.83 ±204.34 μm. Similarly The average thickness ofmyocardium of the female heart in the region of upperleft ventricle was 5344.42± 51.88 μm .the averagethickness of myocardium of the male heart in theregion of middle right ventricle was 5652.4 ± 39.69μm, whereas the average thickness of myocardiumof the female heart in the region of middle rightventricle was 5108.9± 143.11 μm . The averagethickness of myocardium of the male heart in theregion of middle left ventricle was 6231.8 ± 117.86μm Similarly The average thickness of myocardiumof the female heart in the region of middle leftventricle was 5471.23 ± 382.62 μm . The averagethickness of myocardium of the male heart in theregion of lower right ventricle was 5507.47 ± 72.47μm, whereas the average thickness of myocardiumof the female heart in the region of lower rightventricle was 5072.67± 136.59 μm.The averagethickness of myocardium of the male heart in theregion of lower left ventricle was 6503.88 ± 208.93μm. The average thickness of myocardium of thefemale heart in the region of lower left ventricle was5598.05 ± 104.07 μm. These observations were found

statistically highly significant between male andfemale. These observations in the present study werein accordance with the observations noted by Kawelet al. (2012) in human. The difference between theaverage values was statistically significant at 1% andat the same time the mean with at least one commonsuperscript do not differ significantly.

Thickness of pericardium (μm) :

The average thickness of pericardium of the maleheart in the region of upper right Ventricle was1190.23 ± 4.66 μm. Similarly, the average thicknessof pericardium of the female heart in the region ofupper right ventricle was 1188.43 ± 5.39 μm .Theaverage thickness of pericardium of the male heart inthe region of upper left was 1425.73 ± 5.19 μm,whereas the average thickness of pericardium of thefemale heart in the region of upper left was 1333.37±13.61 μm .The average thickness of pericardium ofthe male heart in the region of middle right ventriclewas 1188.45 ± 4.61μm . The average thickness ofpericardium of the female heart in the region of middleright ventricle was 1193.88 ± 5.22 μm .The averagethickness of pericardium of the male heart in the regionof middle left ventricle was 1422.12 ± 5.18 μm,whereas the average thickness of pericardium of thefemale heart in the region of middle left ventricle was1307.26 ± 34.2 μm. The average thickness ofpericardium of the male heart in the region of lowerright ventricle was 1192.03 ± 5.37 μm. Similarly, theaverage thickness of pericardium of the female heartin the region of lower right 1193.88 ± 5.20 μm. Theaverage thickness of pericardium of the male heart inthe region of lower left ventricle was 1421.98 ± 5 .18μm, whereas the average thickness of pericardium ofthe female heart in the region of lower left was1336.98 ± 3.95 μm .These observations werestatistically highly significant between male andfemale sheep. These observations in the present studywere in accordance with the finding reported byHiggins et al. (1986) in human. The differencebetween the average values was statisticallysignificant at 1% and at the same time the mean withat least one common superscript do not differsignificantly.

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Distance between subsequent ‘z’ band (μm) :

The average distance between subsequent ‘z’band of the male heart in the region of upper rightventricle was 4.12 ± 0.82 μm Similarly, the averagedistance between subsequent ‘z’ band of the femaleheart in the region of upper right ventricle was 4.53 ±0.76 μm. The average distance between subsequent‘z’ band of the male heart in the region of upper leftventricle was 6.18 ± 1.39μm, whereas the averagedistance between subsequent ‘z’ band of the femaleheart in the region of upper left was 6.18 ± 1.06 μm .The average distance between subsequent ‘z’ band ofthe male heart in the region of middle right ventriclewas 4.94 ± 0.90 μm. The average distance betweensubsequent ‘z’ band of the female heart in the regionof middle right was 4.53 ± 0.76 μm . The averagedistance between subsequent ‘z’ band of the male heartin the region of middle left Ventricle was 7.41 ±0.90μm The average distance between subsequent ‘z’band of the female heart in the region of upper leftventricle was 6.18 ± 1.06 μm. The average distancebetween subsequent ‘z’ band of the male heart in theregion of lower right ventricle was 4.12 ± 0.82 μm,whereas the average distance between subsequent ‘z’band of the female heart in the region of lower rightventricle was 4.53 ± 0.76 μm .The average distancebetween subsequent ‘z’ band of the male heart in theregion of lower left ventricle was 7.41 ± 0.90μm,similarly the average distance between subsequent ‘z’band of the female heart in the region of lower leftVentricle was 5.35 ± 1.18 μm. These observationswere statistically nonsignificant between male and

female sheep. These observations in the present studywere not in accordance with the observations notedby Robinson et al. (1983) in Mammal.

REFERENCES

Culling, C. F. A. (1969) In “Handbook ofHistopatological technique (including museumtechnique).” 2nd Edn. Pp. 228 – 238 and 477 – 479.Butter worth and Co., Philadelphia and London.

Getty, R. (1975) In “Sisson and Grossman’s ‘TheAnatomy of Domestic Animals” 5th edn. W.B SaundersCompany, Philadelphia.

Higgins C. B., D. Tscholakoff and U. Sechtem (1986)American Journal of Roentgenology.147: 239-244.

Kawel N., E. B. Turkbey , J.J. Carr, J. Eng, A. S.Gomes, W. G. Hundley, C. Johnson , S. C. Masri, M.R. Prince, R. J. van der Geest, J. A. Lima and D. A.Bluemke (2012) Circ Cardiovasc Imaging.5(4):500-8

Masuda H., K. Kawamura, K. Tohda, T. Shozawa,M. Sageshima and M. Honm (1989) . Acta PatholJpn. 39(2):111-20.

Robinson T. F., L. Cohen-Gould and S. M. Factor(1983) A Journal of Technical Methods and Pathology.49(4):482-498.

Snedecor, G. W. and W. G. Cochran (1994) In“Statistical methods” 8th Edn. Oxford and IBHPublishing House, Calcutta.

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Table - 1 : Micrometrical Analysis of different components of ventricle of heart in male &female sheep (Ovis aries) Sr. Parameter Region Side Sex Minimum Maximum Average ± CV F' Value

1 Thickness of endocardium (μm)

Upper Right

Male 21.74 43.48 32.61 ± 3.97

24.83 1.991NS Female 21.74 43.48 28.98 ± 3.62

Left Male 21.74 54.48 38.06 ± 4.67 Female 32.60 54.48 39.87 ± 3.64

Middle Right

Male 21.74 54.48 36.25 ± 4.60

27.054 1.605NS Female 21.74 43.48 32.61 ± 3.97

Left Male 32.60 54.48 43.52 ± 3.99 Female 32.60 54.48 43.52 ± 3.99

Lower Right

Male 21.74 54.48 34.44 ± 5.20

30.383 1.094NS Female 21.74 43.48 32.61 ± 3.97

Left Male 32.60 54.48 43.52 ± 3.99 Female 21.74 54.48 38.06 ± 4.67

2

Thickness of myocardium (μm)

Upper Right

Male 5217.60 5761.10 5416.88b ± 86 13

6.162 21.373** Female 4782.80 5435.00 5036.43b ±

114 58Left

Male 5869.80 7391.60 6521.83a ± 204 34Female 5217.60 5543.70 5344.42b ± 51 88

Middle Right

Male 5543.70 5761.10 5652.4b ± 39 69

4.024 25.811** Female 4782.80 5761.10 5108.9c ±

143 11Left

Male 5869.80 6522.00 6231.8a ± 117 86Female 5217.60 5761.10 5471.23b±82.62

Lower Right

Male 5326.30 5761.10 5507.47b ± 72 47

6.132 17.609** Female 4782.80 5652.40 5072.67c ±

136 59Left

Male 5869.80 7391.60 6503.88a ± 208 93Female 5108.90 5761.10 5598.05b ± 104 07

3 Thickness of pericardium (μm)

Upper Right

Male 1173.90 1206.50 1190.23b ± 4 66

19.279 3.912* Female 1173.90 1206.50 1188.43ab ±

5 39Left

Male 1402.20 1434.80 1425.73a ± 5 19Female 1326.14 1347.80 1333.37a ± 3 61

Middle Right

Male 1173.90 1206.70 1188.45c ± 4 61

3.313 41.010** Female 1173.90 1206.57 1193.88c ±

5 22Left

Male 1402.20 1434.80 1422.12a ± 5 18Female 1137.00 1374.80 1307.26b ± 34 23

Lower Right

Male 1173.90 1206.50 1192.03c ± 5 37

0.912 550.638**

Female 1173.90 1206.57 1193.88c ± 5 20

Left Male 1402.20 1434.80 1421.98a ±

5 18Female 1326.14 1347.80 1336.98b ± 3 95

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Constraints in Adoption of Simple but Vital, Chaff Cutter Technology forEfficient Utilization of Fodder

Sangram Chavan1*, D.S. Deshmukh 1Amol Patil 1 G.R. Gangane 2 Manish Sawant 3

1Department of Veterinary and Animal Husbandry Extension COVAS, Parbhani.2Department of Veterinary Pathology, COVAS, Parbhani.

3Department of Veterinary and Animal Husbandry Extension, Bombay Veterinary College.*Corresponding author e-mail : [email protected]

ABSTRACT

The present study was aimed at finding out the level of constraints faced in adoption ofchaff cutter technology and to identify gap between the recommended technology and its useat farm level. The eight villages of Beed, Latur and Parbhani district of Maharashtra wereselected purposively for the present study by personally interviewing 120 dairy farmers. Theconstraints were categorized in to infrastructural, financial, market related, technical andmanagmental. Referring infrastructure, the respondents expressed lack of awareness oftechnology (82.50%) and subsidy scheme (73.33%) as major constraint. About financialaspects, all of the dairy farmers expressed high cost of machine (100%), followed by highelectricity consumption (95.83%) as the major constraint. All the farmers (100%) expressedno demonstration or trial facility while purchasing followed by no authentic marketinformation of supply (77.50%) about market constraints. Regarding technical constraints,(84.17%) respondents expressed difficulty in regular sharpening of blades followed by(77.50%) poor knowledge of feeding chaffed fodder. As regards managmental constraints,all respondent expressed irregular electricity supply as a major constraints followed byunavailability of regular fodder (83.33%). However, financial support from government orother service providers should be continued with awareness campaign for the poor farmers.Being a proven technology, chaffing needs to be popularized, especially in those areas whereit is not yet fully practiced.

Keywords: Chaffing, Constraints, Dairy Farmers, Feed utilization, fodder

INTRODUCTION

Implementation of any improved scientifictechnology in practical field in all aspects dependson the adoption behaviour of an individual who wantsto implement it. As per the Rogers’s Diffusion ofInnovations (2003), any adopter accepts technologywhen it fulfils the requirement of situation. In animalhusbandry practices, technological gap is a majorproblem in the efforts of increasing milk productionwith minimum feeding cost. Fodder production is thefoundation of a livestock production system while

proper feeding is imperative for achieving high andsustained productivity. Fodder production and itsproper utilization depend on the cropping pattern,climate, socioeconomic conditions, type of livestockand feeding pattern. India with 2.29 % of the landarea of world, maintains nearly 11% of livestock.Majority of livestock are reared on crop residues withlittle supplementation of grains, bran, oil cakes etc.Crop residues and their by-products constitute majoringredients (40%) in daily ration followed by greenfodder (26%), concentrates (3%) and remaining comesthrough grazing. Quantity and quality of feed supply

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remains the major limiting factor to improve livestockproductivity. The livestock population is expected togrow at the rate of 0.55% in the coming years, andthe population is likely to be around 781 million by2050. Though India is among the leading producersof milk, productivity of our animals is 20-60% lowerthan the global average due to improper fodderutilization and due to the lack of scientific feeding ofanimals.

During the last four decades, a number of animal-nutrition-based technologies and practices have beendeveloped with varying degrees of success. Sometechnologies have produced profound beneficialeffects and have been widely used; while others haveshown potential on research stations but have not beentaken up by farmers. Hence there is need to develop atechnology that is suitable for use under localconditions and which is also affordable to farmers.To meet current level of livestock production and theirannual population growth, strategies are needed todesign suitable models for effective and properutilization of fodder in livestock diets. This can beachieved through selection of mechanizational chaffcutter technology which is a key to proper utilizationof fodder, to avoid wastage of fodder and way to caterthe challenges of fodder scarcity. As per reference ofLivestock census 2012, number of mechanical chaffcutter used in livestock sector is India (12534633),Maharashtra (41954) and Marathwada (3076).

General trend in rural area is to offer fodderwithout chopping where as in case of sorghum andmaize they cut it by sickles into large pieces (50 cmlength). In this kind of prevalent practice, wastage ofthe fodder is very high. In order to reduce the wastageof feed resources, the chaffing of fodder waspromoted. Chaff cutter is an agricultural mechanicaldevice which cuts the thick stemmed and un-chewablematerial into small pieces which becomes edible andpalatable to the animals. It reduces selective feeding,increases the intake of animals, improves thedigestibility and conserves energy that they have touse in mastication. It has been estimated that usingchaff cutters, farmers can save 15 to 20% of wastageof fodder. As wastage is reduced, surplus fodderavailable can be fed to more number of livestock.When fodder cultivation was promoted and chaffcutters were introduced in Mahboobnagar district ofAndhra Pradesh, fodder wastage was reduced up to

30 per cent. It appears that use of chaff cutters andchaffing has positive associations with green foddercultivation (Misra et al., 2007). Recognising theimportance of chaff cutters, the Government of Indiain its scheme entitled “Assistance for introduction ofhand- and power-driven chaff cutters” contributes 75per cent and the farmer 25 per cent of the cost of thechaff cutter. Assistance to the tune of five hundredmillion Indian rupees is expected to be given tofarmers to further spread of this simple technology.

Keeping the above problems in view, the presentstudy was taken up with the specific objectives toidentify constraints faced in adoption of chaff cuttertechnology of the said study area by considering theexpectation of rural dairy farmer, cost of production,physiological need of animal, acceptance level oftechnology and role of feeding chaffed fodder etc.

METHODOLOGY

The present study was conducted in Beed, Laturand Parbhani district of Maharashtra state. From allthe 24 villages i.e. 02 villages of four talukas of each03 district were selected randomly. A list of dairyfarmers who were using chaff cutter technology fordairy farming since last three years and havingminimum five dairy animals was prepared and 05respondents were selected purposively from eachvillage. Therefore total size for this study was 120dairy farmers. A pretested semi structured interviewschedule was used to collect the data by personaldialogue method. The questions were related todifferent constraints faced by the farmers whileadoption of simple but worth chaff cutter technology.The data collected through interview was analysedusing statistical tools like the mean, standarddeviation, frequency and percentage for makingsimple comparison.


Constraints imply the problems or difficultiesfaced by dairy farmers with regard to adoption of chaffcutter technology in their dairy enterprise. (Table 1.)

Infrastructural constraints: The majorproblem, faced by the respondent were lack ofawareness of technology use (82.50%) and subsidy

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Table 1 : Distribution of respondents according to the constraints faced while adoption of chaff cuttertechnology

(N=120)

Constraints Frequency Percentage Rank

Infrastructural constraintsInadequate knowledge of technology 68 56.67 IIILack of awareness of technology use 99 82.50 ILess awareness about subsidy scheme 88 73.33 IINeeds based training on safety handling 50 41.67 VIrregular supply of machine 62 51.67 IV

Economic constraintsHigh cost of machine 120 100 IHurdles in getting credit 69 57.5 IIIMore cost of spoiled loose parts 43 35.83 VConsultation charges of local technician 35 29.17 VIMore electricity consumption 115 95.83 IIHigh transportation cost 44 36.67 IV

Technical constraints No technical assistance & guidance 63 52.50 IIIInadequate knowledge of feeding chaffed fodder 93 77.50 IIUnavailability of loose part in local market 43 35.83 VDifficulty in regular sharpening of blades 101 84.17 IUnskilled technician 57 47.50 IV

Market related constraintsIrregularity in prices of fodder 61 50.83 VINo authentic price and supply information 93 77.50 IIOn cash purchasing of machine 49 40.83 VIILess resale value in market 89 74.17 IVNo demonstration/trial facility while purchasing 120 100 INo extra belt, stand and blades with machine 66 55.00 VLess warranty period 45 37.50 VIIINo after sales services 90 75.00 III

Managmental constraintsIrregular electricity supply 120 100 IShortage of man power to take care of machine 73 60.83 VNeed of regular servicing of machine 55 45.83 VIIIAdoptability of chaffed fodder by animal 59 49.17 VIINo availability of fodder round the year 100 83.33 IINeed of shelter for machine protection 59 56.67 VIImmobility of machine 33 65.00 IVTime consuming 44 73.33 III

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scheme (73.33%) followed by inadequate knowledgeof technology (56.67%), irregular supply of machine(51.67%) and need based training on safety handling(41.67%).This might be because of the fact that mostof dairy farmers were satisfied with technology butduring adoption stage , queries regarding technicalknowledge and subsidy scheme raised in their mindmight not be satisfied by the extension worker.

Economic constraints : 100 per cent farmersreported high cost of machine followed by highelectricity consumption (95.83%) as a majorconstraints in dairy farming, while some of the farmersreported constraint to hurdles in getting credit due tounnecessary and irrelevant stipulations of conditions(57.5%), high transportation cost (36.67%), high costof spoiled loose parts (35.83%), high consultationcharges of local technician (29.17%). The probablereason might be that in present study herd size offarmer was medium, but still they have to invest hugeamount without loan for machine purchasing, alsomechanizational small capacity machine was notavailable in market.

Market related constraints : All the farmersexpressed no trial facility while purchasing and noauthentic market information of supply (77.50%) asa major constraints while some expressed high pricesof machine (75%), lack of after sales services (17%),less resale value in market (74.17%), no extra belt,stand and blades with machine(55%), irregularity insupply and prices of fodder (50.83%), on cashpurchasing of machine (40.83%), less warranty period(37.50%) . Probable reason could be that the price,supply and demand of fodder might be controlled bymany factors which are not in control of dairy farmer.Lack of coordination between farmers and extensionworkers and involvement of commercial agenciesmake the technology more expensive for farmers.

Technical constraints : The majority ofrespondents expressed difficulty in regular sharpeningof blades (84.17%) and inadequate knowledge offeeding chaffed fodder (77.50%) followed by notechnical assistance & guidance (52.50%), unskilledtechnician (47.50%), unavailability of spoiled, loosepart in local market (35.83%) as a constraint.

Managmental constraints : 100 per centrespondent reported no constant electricity supplyfollowed by unavailability of fodder round the year(83.33% while some reported constraint to the factorof time consuming(73.33%), immobility ofmachine(65%), shortage of man power to take careof machine(60.83%), need of shelter for machineprotection (56.67%), adoptability of chaffed fodderby animal (49.17 %), irregular visits of localmechanic, need of regular servicing of machine(45.83%). Similar findings were previously reportedby Sagari (2001), Misra et al., 2007, Rathod (2011).The use of chaff cutters and chaffing has positiveassociations with green fodder cultivation but inMarathwada region of Maharashtra, due to low rainfall condition; cultivation of green fodders is notwidespread. Hence there is a need to guide the dairyfarmers about fodder cultivation.

CONCLUSION

From the above results, it can be concluded thatlack of awareness of technology use with subsidyscheme, inadequate knowledge and irregular supplyof machine was major constraints. High cost ofmachine followed by inadequate money and lack ofloan facility with high electricity consumption werea major constraints regarding financial aspect. No trialfacility while purchasing of machine and no authenticmarket information of supply, prices of machine werethe problems faced by the respondents in the market.The majority of respondent opined difficulty in regularsharpening of blades, poor knowledge of feedingchaffed fodder and unavailability of regular fodderwith irregular electricity supply as major constraints.

Based on the study following recommendationsare made.1. To increase rate adoption, technology should be

commercially diffused by assessing its perceivedattributes and production factor (man, money andmaterial, labour) with increase in incentives fromthe government agencies and cooperatives whichwill defiantly give impetus to its adoption.

2. In developing countries like India, to enhancelivestock productivity, fodder cultivation andchaffing technology should be encouraged andpromoted by organizing extension awareness

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program by demonstrating use of chaff cuttersand its advantages together so that this simple,but vital technology become popularized at a fastpace.

3. Farmer-scientist-extension-worker interactions,participatory technology appraisal methods,exposure visits and need based training programwhich are often quite effective, should bepromoted.

4. The technology needs to be refined further,technically and made simpler for the farmer sothat larger farmers may come forward and adoptthis technology.

REFERENCES

Chander M. (2011) Chaff cutters and fodder chaffing:A simple technology worth Adoption. Successes andfailures with animal nutrition practices andtechnologies in developing countries. FAO Electronic

Conference, 1-30 September 2010, Rome, Italy,Proceedings 11 (133-36).

Pareek P. (2014) A Study on Market DevelopmentAssistance to Samalkha (Haryana) Chaff CutterIndustrial Cluster To Enter In Gujarat Market. PacificBusiness Revi International 6: 10-12.Misra, A.K, Rama Rao, C.A. Subrahmanyam, K.V.,Vijay Sankar, B.M., Shivarudrappa, B. &Ramakrishna, Y.S. (2007) Strategies for livestockdevelopment in rain fed agro-ecosystem of India.Livestock Res. Rural Develop. 19:83-85.

Misra, A.K. (2010). Analysis of potentials andproblems of dairy production in rain fed agro-ecosystem of India. Indian J. Anim. Sci., 80 (11):1126–1133.

Rathod P. (2011). Socio-personal profile andconstraints of dairy farmers, Karnataka J. Agric. Sci.,24 (4): 619-621.

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J. Bombay Vet. College (2014) 21(2) Short Communication

Inguinal hernia associated with cryptorchidism in a Pug

S.V. Upadhye, N.P. Dakshinkar, G.R. Bhojne, A.P. Gawande, G.S. Khante, A.R. Ratnaparkhi and Nitya NirodyTeaching Veterinary Clinical Complex, Nagpur Veterinary College, Nagpur

Maharashtra Animal & Fishery Sciences University, Nagpur.

Cryptorchidism is the condition in which eitheror both the testicles fail to descend into the scrotumafter birth. In dogs, this condition has been reportedmore commonly in toy breeds than in the larger breeds(Pendergrass and Hayes, 1975). Inguinal hernias areprotrusions of organs or tissues through the inguinalcanal adjacent to the vaginal process (Fossum, 2007).Such hernias occur as a result of a defect in theinguinal canal allowing the abdominal contents toprotrude through it (Smeak, 2003). The commoncontents of inguinal hernia are omentum, fat, ovary,uterus, small intestine, colon, bladder and spleen(Bellenger, 1996). The delay in descent of testiclesinto the scrotum causes the inguinal canal to remainwide open for a longer duration. Cryptorchidism isthereby commonly associated with congenital inguinalhernia.

A male Pug aged about 10 months was presentedto TVCC, Nagpur Veterinary College with a slowprogressive swelling at left inguinal region (Fig.1).On physical examination, the swelling was found tobe non-painful and irreducible. A thoroughexamination revealed that the dog was monorchid.Differential diagnosis included neoplasia of the

retained testicle or irreducible inguinal hernia. Theage of the patient was significantly lower than themean age of susceptibility to testicular neoplasia.Ultrasonography was performed to identify thestructure involved in the inguinal swelling. Omentumwas observed along with the borders of the hernialsac and testicular structure. Based on the history,physical examination and ultrasonography, thecondition was diagnosed as a congenital inguinalhernia and surgical correction was advised due to thepossibility of incarceration of intestinal loop withinthe hernia in future.

The dog was prepared for surgery and the leftinguinal region along with the entire ventral abdomenwas prepared for aseptic surgery in routine manner.Premedicants were administered and the dog wassedated with Xylazine at 1mg/kg body weight.Dissociative anesthesia was induced and maintainedwith Ketamine at 5mg/kg body weight and Diazepamat 0.5mg/kg body weight. An elliptical incision wastaken over the hernia through the skin andsubcutaneous tissue. The hernial sac was exteriorizedby blunt dissection. After failed attempt to reduce thehernia manually, the hernial sac was incised carefullyto expose the hernial contents. Omentum was foundwithin the hernial sac along with the undescendedtestis and spermatic cord. This testis was noted to besmaller and atrophied (Fig.2). The undescendedtesticle was self-ligated and removed through theinguinal incision. The inguinal canal was enlarged andthe omentum replaced into the abdominal cavity afterdissecting the adhesions. The hernial ring was closedwith No. 0 Vicryl. The subcutaneous tissue wassutured using No. 1-0 Vicryl and the skin incision wassutured using nylon. The descended testicle wasremoved through a prescrotal incision. Postoperativetherapy included Ceftriaxone @ 20mg/kg body weightfor five days and Meloxicam @ 0.5 mg/kg bodyweight for three days. The dog recovered uneventfullyand no recurrence of hernia was reported. Sutures wereremoved on 10th day postoperatively.

Fig. 1 : Swelling at the left inguinal canal

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DISCUSSION

Unilateral cryptorchidism shows higherincidence than bilateral cryptorchidism in dogs withthe right testicle being more commonly undescended.Also, the undescended testis is more commonlylocated in the abdomen than in the inguinal canal(Boothe, 2003). However, in this case, the left testiclewas noted to be undescended and located in theinguinal canal. Inguinal hernias are broadly classifiedas congenital and acquired hernias. Congenitalinguinal hernia occurs more commonly in male dogs(Smeak, 2003). Inguinal hernia and cryptorchidismhave been reported to be inheritable traits (Burke andReynolds, 1993). Also, the risk of testicular neoplasiain a cryptorchid dog is upto 13.6 times higher than ina normal dog (Hayes and Pendergrass, 1976). In viewof these reports, castration was performed duringherniorrhaphy in this case to prevent furthertransmission of the inheritable traits and as aprophylactic measure against testicular neoplasia. Thecommon complications of inguinal hernia repair

Fig. 2 : Omentum and atrophied testicle as the hernia content

include hematoma or seroma formation, incisionalinfection, wound dehiscence, hernia recurrence,peritonitis, sepsis and death (Pratschke, 2002).However, the patient in this case recovereduneventfully with no complications.

REFERENCES

Bellenger, C. R. (1996). Inguinal and scrotalherniation in 61 dogs. Aust. Vet. Prac. 26: 58-59.

Booth, H. W. (2003). Testes and Epididymis. InTextbook of Small Animal Surgery 3rd edn. Slatter,D. W. B. Saunders, Philadelphia, pp 1524-1527.

Burke, T. J. and H.A. Reynolds (1993). The testis. Intextbook Disease Mechanisms in Small AnimalSurgery by Bojrab M. J., 2nd edn. Lea & Febiger,Philadelphia, p 545.

Fossum, T. W. (2007). Surgery of the abdominal cavityin Textbook Small Animal Surgery by Fossum, T. W.3rd ed. Mosby Elsevier, Philadelphia, pp: 317-338.

Hayes H. M. and T.W. Pendergrass (1976). Caninetesticular tumours: epidemiological features of 410dogs. International Journal of Cancer 18(4): 482-487.

Pendergrass, T. W. and H.M. Hayes Jr. (1975).Cryptorchidism and related defects in dogs:epidemiologic comparisons with man. Teratology.12(1): 51-55.

Pratschke, K. (2002). Management of hernias andruptures in small animals. In Pract. 24: 570-581.

Smeak, D. D. (2003). Abdominal hernias. In Textbookof Small Animal Surgery, by Slatter D. 3rd edn.Saunders, Philadelphia. Pp. 452-455.

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A five year old sheep was presented to theDepartment of Veterinary Surgery and Radiology,

Bombay Veterinary College with a history of anautomobile accident leading to lameness of leftforelimb. Lateral and anterio-posterior radiographrevealed complete oblique fracture of mid shaft ofleft humerus. It was decided to fix the fracture byintramedullary pinning. The patient was anaesthetizedwith Ketamine hydrochloride @ 22mg/kg bodyweight IM and Xylazine hydrochloride @ 0.2mg/kgbody weight IM and fluid therapy was given till theend of surgical procedure.

The patient was prepared for routine asepticsurgery. The shaft was approached from anterio-lateralaspect by a skin incision on a line joining the lateraltuberosity and the epicondyle. A Steinmann pin of7mm in diameter was introduced in a retrogrademanner by use of a chuck followed by reduction offracture. Further, the Steinmann pin was retractedpartially from the proximal extremity and inserted innormograde manner through proximal extremity ofthe bone. It crossed the fractured portion and reachedupto the distal extremity. An ancillary cerclage wiringwas done at the fracture site (Fig.1). The soft tissuealong with periosteum was closed in a simple

Retrograde intramedullary pinning for repair of humerus fracture in a sheep

M. S. Rajhans, D. U. Lokhande, G. S. Khandekar and S. V. GaikwadDepartment of Veterinary Surgery and Radiology


continuous fashion using 1-0 chromic catgut. The skinwound was sutured in a routine manner. Ceftriaxonesodium @ 10mg/kg body weight intramuscularly oncedaily was administered for 7 post-operative days.Flunixin meglumine @ 1.1mg/kg body weight IM wasadministered once daily for 3 post-operative days.

On sixth post-operative day, the patient startedbearing partial weight on affected limb. By 20th post-operative day, complete weight bearing was noted onthe affected limb. The skin sutures were removed on10th post-operative day. The functional limb usageby the end of 45th post-operative day was good.

Chawla et al. (1983) conductedhistomorphological studies of tibial fracture healingin sheep, immobilized with external coaptation,intramedullary pining and plating. They demonstratedthat bony union of fragments was achieved within 9

Fig. 1 : Stabilization of fractured fragments with Steinmannintramedullary pin and cerclage wiring at fracture site

Fig. 2 : Intramedullary pinning in complete oblique fractureof mid shaft of humerus

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weeks in pinning and plating, with little calcification.They concluded that fixation occurred more rapidlyin intramedullary pining than in plating. St-Jean et al.(1992) used intramedullary pin with cerclage wiresand external fixator for the repair of a comminuted,spiral oblique, proximal diaphyseal femoral fracturein a 7-day-old calf. They reported good healing at thefracture site on 45 days after surgery and the calf hadnormal gait eight months after the surgery. Awatif etal. (2006) concluded that Kirschner intramedullarypinning can be used successfully in immobilizationof femoral fracture in goats. There is no apparentdifference in results of oblique or transverse fracturehealing. Avasthi et al. (2012) reported a completeoblique fracture of humerus in a goat which wasimmobilized and fixed by intramedullary pinningalong with wiring. The goat recovered uneventfullyin due course of time.

In present case, intramedullary pinning was goodresulting into early ambulation of limb withprogressive fracture healing.

REFERENCES

Avasthi, H. A., Patel, J. B. and Patel, P. B. (2012).Surgical management of mid-shaft humerus fracturewith intramedullary pinning in a goat. Intas Polivet.13(2): 380-381.

Awatif, M. E, Ali, O. S. and Shnain, H. M. (2006).Kirschner Intramedullary Pinning for FemoralFracture in Caprine. Surgery Journal. 1(2-4): 75-77.

Chawla, S. K., I. S. Chandna, K. C. Bhatia and A. P.Singh (1983). Histomorphological studies of tibialfracture healing following internal and externalimmobilization in sheep. HAU J. Res., 13 (1):11-18(Vet.Bull 1984, Abstr.224).

St-Jean G., R. M. De Bowes, A. M. Rashmir and T. J.Engelken (1992). Repair of a proximal femoralfracture in a calf using intramedullary pinning,cerclage wire, and external fixation, J. Am. Vet. Med.Assoc., 200: 1701- 03.

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Monster is an individual having multipleanomalies and involving many organs and systemsof body. The causes of anomalies are not alwaysknown, most of them have their origins within thefirst few weeks after fertilization. However, some maydevelop later in pregnancy. Many monsters even havetheir inception in the germplasm. These are hereditaryMendelian dominant or recessive genes responsiblefor monstrocities. Others are due to environmentalinfluences, such as exposure to teratogenic drugs orviral infection during pregnancy (Roberts, 1971). Thisreport deals with unrecorded deformities in a cowfoetus.

Case History and Clinical Observations

A five year old Deoni cow in her second gestationwas presented at Reproductive Disorder ManagementCell, Teaching Veterinary Clinical Complex,Veterinary College, Udgir with history of completionof gestation period, continuous labor pains,restlessness, and straining since last 12 hours. TheChorioallontoic sac was rupture 6 hours before thecase was presented. The cow was dull, depressed, andexhausted due to severe straining. One hind limb wasprotruding from the vulva. Thorough Gynaeco-clinicalexamination was carried out to diagnose the cause ofDystokia. The foetus was in posterior longitudinalpresentation and dorso-lumbar position. Foetalmovements and other reflexes were absent. The deadfoetus was removed with Forced Traction andobstetrical mutations, after ample lubrication whichwas abnormal (Fig. 1 and 2).

Before manipulation, the animal was treated withDextrose 5% @ 3000ml I/V, cortisone and antibiotics.Epidural anaesthesia was performed with 2 %lignocaine hydrochloride. Birth canal was lubricatedwith carboxy methylcellulose sodium @ 20 gm perlit. in luke-warm water. The foetus was repelled and

Dystokia due to monster with multiple bone deformities in a Deoni cow

U. B. KumbharReproductive Disorder Management Cell

Department of Animal Reproduction, Gynaecology and ObstetricsBombay Veterinary College Parel Mumbai - 400 012.

Corresponding author E mail : [email protected]

corrected by following all aseptic precautions. Withthe help of obstetrical snare forced traction was appliedand a dead foetus was delivered.

The necropsy findings of foetus revealed welldeveloped brain, ribs were absent, spinal cord absencewith no optimum development of bones of vertebralcolumn. Right foreleg was ill developed (micromelia).Radius ulna (elbow joint) and metacarpal bones werealso ill developed (perodactyly), knee joint of rightside was not properly developed and appeared shortas compared to that of the knee joint of left side(micromelia). Right hind leg was having defectiveposture (micromelia). Tail and anus were properlydeveloped and all visceral organs were ill developed.

Treatment and Discussion

The fetus revealed contractures and ankylosis ofall the joints with variable degree of flexion andextension (Figure 1) and hence termed asarthrogryposis. Mahajan et al., (2006) also describeda case of arthrogryposis in a buffalo calf causing

Fig. 1 : Dead foetus removed with forced traction

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dystocia. In the present case the flexion of jointsresulted in the dystocia. Such types of monsters aregenerally considered to be due to a simple, autosomalrecessive defects with some modifiers (Roberts, 1971).Monster with multiple bone deformities is thoughuncommon have been reported by (Sastry, 1983) sameas this monster in which ribs were absent, there wasno proper development of bones of vertebral columnand spinal cord was absent. Right foreleg was illdeveloped (micromelia). Radius ulna (elbow joint) andmetacarpal bones ill developed (perodactyly), kneejoint of right side was not properly developed and

appeared short as compared to that of the knee jointof left side (micromelia). Right hind leg was havingdefective posture (micromelia). Tail and anus wereproperly developed and all internal organs were ill

formed. The fixation of the joints may have been dueto lack of extensibility of the muscles, ligaments oratrophy resulting from neuropathy (Tyagi and Singh,1996). Non-hereditary causes of muscularcontractures have also been reported (Roberts, 1971).

REFERENCE

Mahajan, A. Sathya Singh, A. P. and Prabhakar, S.(2006). A case of arthrogryposis in a buffalo calfcausing dystocia. Indian J. Anim. Reprod., 27: 86-87.

Roberts, S. J. (1971). Veterinary Obstetrics andGenital Diseases, 2nd Edition CBS Publishers andDistributors, Delhi. pp. 51, 70-73, 277-278.

Sastry G. A. (1983). Veterinary pathology, 6th EditionCBS Publishers and Distributors, Delhi. pp 462.

Tyagi, R.P.S. and Singh, J. (1996). Ruminant Surgery.CBS Publishers. New Delhi, pp. 200

Fig. 2 : Monster with multiple bone deformines removedwith forced traction

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Cranial Neoplasia in a Dog

S.V. Upadhye, S.B. Akhare, P.T. Jadhao, G.S. Khante, Shalaka Salvekar, S.S. Pitlawar and Nitya NirodyTeaching Veterinary Clinical Complex, Nagpur Veterinary College, Nagpur

Maharashtra Animal & Fishery Sciences University, Nagpur

Neoplasia of the skeletal system is lesscommonly encountered in dogs in comparison to thatof skin, genital, gastrointestinal and hematopoieticsystems (Hardy et al, 1967). Long bones are the mostcommon sites of primary bone neoplasia in dogs(Brodey and Riser, 1969). Within the axial skeleton,neoplasia of the cranium is a rare occurrence withhigher incidences of mandibular and maxillaryneoplasia (Heyman et al, 1992).

A male German shepherd cross breed dog aged14 years was presented at Teaching Veterinary ClinicalComplex, Nagpur Veterinary College with abnormalslow growing swelling over the right side of theforehead (Fig.1). The owner reported normalorientation and activity. On physical examination, ahard painless mass was palpable extending from overthe frontal, temporal and occipital cranial bones.Neurological examination was performed to assessthe cranial nerve reflexes. No deficits were noted inthe neurological examination. Radiography wasundertaken to identify the structures involved. A rightlateral radiograph of the skull revealed periostealelevation with a radio-opaque mass proliferating fromthe temporal and occipital bones (Fig.2). A lateralthoracic radiograph did not reveal any metastaseswithin the thoracic cavity. Further ultrasoundexamination did not reveal any metastatic lesions in

either spleen or hepatic parenchyma. Based on thehistory, clinical examination and the “popcorn ball”appearance of the lesion on radiograph (Boston,2010), the mass was tentatively diagnosed asMultilobular Osteochondrosarcoma (MLO). The lackof clinical symptoms was consistent with the findingsof other researchers who reported that MLO involvingthe skull may not always be accompanied byneurological signs or deficits (Lipsitz et al, 2001).

The options of surgical excision andchemotherapy were suggested to the owner. However,based on the risks explained pertaining to eachprotocol, the owner refused treatment due to lack ofobvious distress to the animal and economicconstraints. Further, the case was lost to follow up.

DISCUSSION

On account of the poor prognosis and high riskinvolved, treatment of neoplasia involving cranialbones is rarely initiated. Aggressive surgical excisionhas been shown to improve mean survival time fromdiagnosis (Dernell et al, 1998). MRI and CT imagingprior to surgery for assessing cortical collateralcirculation helps to gauge the prognosis in cases withpossibility of bilateral disruption of transverse venoussinuses following tumour resection (Gallegos et al,2008). The defect formed by the partial craniectomycan be covered by the temporalis muscle when there

Fig. 1 : Hard swelling over the skull Fig. 2 : Lateral radiograph showing the osteogenic tumour

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is no or minimal invasion of this muscle by the tumour.More advanced techniques include cranioplastyfollowing en bloc resection. Utilization of autologousbone dust has been reported to obtain a complete bonycovering of the skull defect (Shehadi, 1970).Cranioplasty can also be performed by utilizing boneallograft or polymethylmethacrylate implant(Moissonnier et al, 1997). Radiation therapy with orwithout chemotherapy is an alternative to surgicalexcision. In this case, the extent of local spread of theneoplasia was large and thereby, the feasibility ofsurgical excision was doubtful. Economic constraintsprevented the utilization of CT and MRI as prognosticindicators for surgical intervention. Ownercompliance is also an important part of decidingtherapeutic protocol for such conditions.

REFERENCES

Boston, S. E. (2010). Craniectomy and orbitectomyin dogs and cats. Can Vet J. 51(5): 537–540.

Brodey, R. S. and W.H. Riser (1969). Canineosteosarcoma: A clinico-pathologic study of 194 cases.Clin Orthop 62:54.

Dernell, W. S.; R.C. Straw; M.F. Cooper; B.E. Powers;S.M. LaRue and S.J. Withrow (1998). Multilobular

osteochondrosarcoma in 39 dogs: 1979-1993. J AmAnim Hosp Assoc, 34: 11-18.

Gallegos, J.; T. Schwarz and J.F. McAnulty (2008).Massive midline occipitotemporal resection of theskull for treatment of multilobularosteochondrosarcoma in two dogs. J Am Vet MedAssoc. 233(5):752-7.

Hardy, W. D. Jr., R.S. Brodey and W.H. Riser (1967).Osteosarcoma of the canine skull. J Am Radiol Soc8:5

Heyman, S. J.; D.L. Deifenderfer; M.H. Goldschmidtand C.D. Newton (1992). Canine axial skeletalosteosarcoma. A retrospective study of 116 cases(1986 to 1989) Vet Surg.; 21: 304–310.

Lipsitz, D.; R.E. Levitski, and W.L. Berry (2001).Magnetic resonance imaging features of multilobularosteochondrosarcoma in 3 dogs. Vet RadiolUltrasound, 42: 14-19.

Moissonnier P.; Devauchelle, P. and Delisle, F. (1997).Cranioplasty after en bloc resection of calvarialchondroma rodens in two dogs. J Small Anim Pract,38: 358-363.

Shehadi, S. I. (1970) Skull reconstruction with bonedust. British Journal of Plastic Surgery, 23: 227-234.

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Mastocytoma in a dog - A case report

M.S. Rajhans, G.S. Khandekar , D.U. Lokhande, S.V. Gaikwad and D.J. PatilDepartment of Surgery and Radiology, Bombay Veterinary College, Parel, Mumbai - 400 012

An eight year old Great Dane, male dog waspresented to the department of Surgery and Radiology,Bombay Veterinary College with the history of a largenodular growth in the left forelimb paw in theinterdigital space which was increased in size over aperiod of weeks to that extend that the dog was unableto bear weight on that limb and hence was limping.On clinical examination, there was ulceration andoedematous swelling. The growth was multilobate,medium sized and irregular in shape which wasobliterating the interdigital space and covered by skinexternally (Fig. 1). Hence, surgical intervention wasplanned for. The complete blood counts were alsofound within normal range.

The surgical site was prepared aseptically. Theanimal was premedicated with atropine sulphate @0.04 mg/kg subcutaneously followed bytriflupromazine hydrochloride @ 1 mg/kg bodyweight intravenously. General anaesthesia wasinduced with thiopentone sodium @ 10 mg/kgintravenously. Anaesthesia was maintained with 1/3to 1/2 of the calculated dose as and when required.An elliptical incision was taken at the base of tumorand the soft tissue growth was separated slowly from

the surrounding healthy tissue (Fig. 2). The bleedingwas arrested by ligating the major vessels withchromic catgut no.0 and the skin was sutured in asimple interrupted manner using black braded silkno.0. The growth was preserved in 10% formalinsolution and was sectioned for histopathologicalexamination.

Postoperatively, cefotaxime sodium was given@ 20 mg/kg b.wt. i.v., twice a day for 5 days and Inj.Meloxicam @ 0.2 mg/kg b.wt. s.c., once a day wasalso given daily for 3 days. A protective pressure pawbandage was also applied and the sutures wereremoved on the 10th post-operative day.

Histopathological examination of the tissuerevealed, moderate accumulation of round, oval topolygonal cells of medium size in dermis. The cellsshowed variable amount of faint eosinophiliccytoplasm with indistinct cell boundaries. The nucleiwere centrally placed, round or oval with distinct

Fig.1 : Multilobate nodular growth obliterating theinterdigital space

Fig. 2 : Separation of soft tissue growth

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nuclear membrane. The chromatin granules weremedium to coarse and usually prominent towardsperiphery of the nucleus, nucleoli were inconspicuous.The cells lacked any specific architectural pattern andwere spread loosely in discrete clusters. Connectivetissue fibrosis and laid down collage formed majorpart of the mass. The histopathological features weresuggestive of mastocytoma (Fig. 3).

Misdop (2004) reported that mast cell tumors(MCTs) are highly invasive and metastatic and arethe most frequent round cell tumors in dogs,

Fig. 3 : Histopathological examination suggestive ofmastocytoma

comprising 16–21% of all cutaneous tumorsdiagnosed and are the most common skin tumors indogs representing about 7% to 21% of all dog skintumors and 11% to 27% of malignant skin tumors inthis species. Welle et al. (2008) concluded that canineMCTs have variable biologic behaviors, ranging fromsolitary benign masses that can be cured with surgeryalone to systemic and potentially fatal metastaticdisease and are always considered potentiallymalignant, but their true metastatic potential is notentirely known. MCTs are most often gradedhistologically by the scheme described by Patnaik etal. (1984). Tumours of grade I are circumscribed,mainly dermal in location, and consist of well-differentiated mast cells with prominentmetachromatic cytoplasmic granules. Poor cellulardifferentiation, aggressive growth pattern, moderateto high mitotic activity, mitotic atypia and sparse toabsent cytoplasmic granulation are the major featuresof grade III tumours. Grade II MCTs constitute anintermediary form between the two other grades, butthey tend to infiltrate more deeply than grade Ineoplasms. MCTs are routinely diagnosed by cytologyand histopathology.

REFERENCE

Misdorp, W. (2004). Vet. Quarterly 26:156-169.

Patnaik, A. K., Ehler, W.Y. and MacEwen, E.G.(1984). Vet. Pathol. 21: 469-474

Welle, M. M., Rohre Bley, C., Howard, J. andRufenacht, S. (2008). Vet. Dermatol. 19: 321-339.

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Anaesthetic and Surgical Management of Severe Traumatic Wound on the Tail in IndianPalm Squirrel (Funambulus Palmarum).

Aakash Vedpathak, K.S. Chaudhari, D.U. Lokhande, G.S. Khandekar, S.D. Tripathi,Maneesh Tiwari, Niharika Sawant and Tenzing Sherpa

Department of Surgery & Radiology,Bombay Veterinary College, Parel, Mumbai - 400 012.

The Indian Palm Squirrel (Funambuluspalmarum) also known as three-striped palm squirrelis endemic to southern India and Sri Lanka (Molur etal. 2005). Wild small mammals such as squirrels areoften brought into veterinary clinics with traumaticwounds. The proper restraint of these animals isessential for adequate and complete treatment. Wildsquirrels are usually timid and resist manual handling.

Due to severe traumatic injuries to the tail, bloodsupply and nerve supply to the part which is distalfrom the wound is hampered. This may lead toischemia and necrosis of a portion of the tail. In suchsituation it is often recommended the tail of the animalbe amputated at a higher level to allow quicker healingand prevent spread of localized infection and necrosis.

Case history and Observations

An adult female Indian palm squirrel(Funambulus palmarum), weighing 100 gms waspresented to the surgical ward of Bai SakarbaiDinshaw Petit Hospital for Animals, Parel, with ahistory of a traumatic wound at the tail atapproximately 1/3rd length of the tail. On physicalexamination it was found that the part of the tail distalto the wound was cold to touch, showed no movementand the animal did not show any pain reflex whenpinched at that area. Part of the coccygeal vertebraeat that area were exposed. Hence, based on clinicalexamination it was decided that the part of the taildistal to the wound was not viable and the decision toamputate the tail at a higher level was taken.

Treatment and discussion

The squirrel was restrained using a cloth (Arenz,1997 & Lin, 2007) so that the thigh region wasexposed. Anaesthesia was induced with Inj. Ketamine@ 75 mg/kg and xylazine @10 mg/kg given intramuscularly (Olson, 1986) in the thigh muscles usingan insulin syringe with a 25 gauge needle. The site ofoperation was prepared aseptically. The animal was

placed in sternal recumbency. The ring blocktechnique was used to infiltrate 2% lignocaine at anarea proximal to the operative site and a tourniquetwas tied at the base of the tail. U shaped incisionswere taken on the ventral and dorsal aspect of the tailat a level proximal to the wound. The muscles weredissected and the distal portion of the tail wasamputated. Minor bleeding was controlled usingelectro-cautery. The skin was sutured routinely usingsimple interrupted sutures. The animal wasadministered amoxicillin orally @ 25mg/kg bodyweight postoperatively. The animal made anuneventful recovery from anaesthesia. The surgicalwound healed 10 days postoperatively and animal wasdischarged.

SUMMARY

A case of anaesthetic management and tailamputation in an Indian palm squirrel (Funambuluspalmarum) has been reported.

REFERENCES

Arenz C.L. (1997). Handling of fox squirrels:ketamine hydrochloride versus simple restraint.Wildlife society bulletin, 25(1): 107-109.

Hui-Chu Lin, Dissociative Anesthetics (2007). In:Thurmon JC, Tranquilli WJ, Benson GJ, eds. Lumb& Jones Veterinary Anesthesia and Analgesia. 4th ed.Baltimore: Lippincott Williams & Wilkins: 301-353.

Molur S. et al (2005). Status of South Asian Non-volant Small Mammals: Conservation Assessment andManagement Plan (C.A.M.P.) Workshop Report ZooOutreach Organisation/ CBSG-South Asia,Coimbatore, India, 618pp.

Olson ME, McCabe K. (1986). Anesthesia in theRichardson’s ground squirrel: Comparison ofketamine, ketamine and xylazine, droperidol andfentanyl, and sodium pentobarbital. J Am Vet MedAssoc 189:1035.

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Reconstruction of buccal mucosa in a dog

S.V. Upadhye, S.B. Akhare, M.S. Dhakate, G.S. Khante, A.P. Gawande and Shalaka SalvekarTeaching Veterinary Clinical Complex, Nagpur Veterinary College, Nagpur

Maharashtra Animal & Fishery Sciences University, Nagpur

Injuries of the oral cavities are common in canineowing to their scavenging, sniffing and biting habits.They are likely to take up foreign bodies and inanimateobjects due to curiosity. The foreign objects such assharp objects or explosives, if accidentally chewedby them may lead to serious consequences. A case ofsuch buccal injury and its surgical management isreported.

A Pakistani Bully dog of eighteen months agewas presented to TVCC, Nagpur Veterinary Collegewith a day old traumatic oral injury on account ofexplosion of a crude bomb accidentally chewed bythe dog on the farm. The explosion of crude bombresulted in severe injury and profuse haemorrhagefrom the oral cavity. The hemorrhage was controlledby application of ice and pressure packs by the ownerfollowing the trauma. Preliminary clinicalexamination of the oral cavity revealed an irregular Yshaped vertical wound over the right upper labia alongwith extensive swelling, submucosal damage andischemia of the labia and cheek.

The dog was sedated with intramuscularadministration of Xylazine at 1mg/kg body weightand dissociative anesthesia was induced withKetamine (5mg/kg body weight) and Diazepam(0.5mg/kg body weight) intravenously. A thoroughphysical examination of the oral cavity revealed severetrauma to the buccal mucosa along with intensecongestion, ischemia and oedema (Fig. 1). Theexamination also included right lateral, open mouth

and dorsoventral radiographs to assess the damage tothe maxilla and mandible. No fractures were identifiedon examination of the radiographs.

The buccal cavity was lavaged thoroughly withwarm sterile isotonic saline. The ischemic and necroticareas of the buccal mucosa were identified andresected. Reconstruction of the viable buccal mucosawas planned to close the large defect. The alveolarmucosa which was separated from the gingiva at thelevel of third and fourth premolar was opposed andsutured to it using 3-0 chromic catgut. The defect inthe buccal mucosa was corrected in inverted Y-patternwith simple interrupted sutures using 3-0 chromiccatgut. The upper labial skin defect was closed bysimple interrupted sutures using monofilament nylon(Fig. 2). An Elizabeth collar was advised to be usedto prevent self mutilation of sutures. The dog wasadvised to be kept off feed for five days followed bygradual introduction of soft diet. Intravenous fluidtherapy was administered for 7 days along withparenteral Ceftriaxone. Meloxicam was administeredparenterally for 3 days.

The dog showed recovery with reduction inoedematous labial swelling and return to normalactivity. However, disruption of external upper labialskin sutures occurred by self mutilation on secondpostoperative day. The dog was sedated andanesthetized following the same protocol. Thedisrupted sutures were removed and resuturing wascarried out in similar pattern. This was followed by

Fig. 1 : Severe injury to oral mucosa following crude bombexplosion

Fig. 2 : Reconstructed oral mucosa

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uneventful recovery with return to normal feedingwithin 10 days.

Reconstructive surgery of oral cavity primarilyaims at restoration of function and adequate cosmeticoutcome. Primary closure of oral mucosal defects isfeasible on account of the rich blood supply to theregion and the resistance to infection provided by theantimicrobial activity of saliva (Radlinsky, 2013). Oralmucosa is preferably apposed and sutured witheverting suture pattern to counter the inward curlingof the edges and to bring about apposition ofsubmucosa to submucosa to enhance healing. Verticalmattress suture is the most preferred suture patternfor oral mucosa due to the eversion of the edges and

its minimum impact on circulation at wound edges(Pavletic, 2010). However, due to paucity of mucosalflap after resection of the necrotic tissue, verticalmattress sutures could not be applied in this case.However, the methodology adopted here in this caseresulted into good outcome.

REFERENCESPavletic M. M. (2010). In Atlas of Small AnimalWound Management and Reconstructive Surgery, 3rd

ed. Wiley-Blackwell, p512.

Radlinsky M.G. (2013) In Small Animal Surgery, 4th

ed. Elsevier Mosby, p387-389.

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Mumbai is a metropolitan city with a largepopulation of stray dogs. Automobile accidents arecommon and usually involve trauma to the soft tissueand many a times also involving the skeleton. Facialtrauma involving the maxilla and mandibles usuallyrenders such animals incapable of taking food or liquidand may lead to starvation and even death if not treatedpromptly. The use of miniplates and screws has beenrecommended for repair of fractures of the mandibleinvolving the body of the mandible and the angle ofthe mandible (Boudrieau, et al., 1996).

Case history and Observations

A male, non descript dog weighing 15 kilogramswas presented to the Bai Sakarbai Dinshaw PetitHospital for Animals, Parel with a history of anautomobile accident. On clinical examination severalwounds were seen on the animals head. The animalwas found to be severely dehydrated. Mild purulentdischarge was seen at the wounds. Halitosis withmucoid foul smelling saliva was seen in the oral cavity.The dog was unable to close its jaws. On palpation itwas discovered that the dog had suffered fractures onthe mandibular body on both right and left mandibles.Radiography revealed the presence of an obliquefracture on the right mandible at the horizontal bodyof the mandible. The fracture of the left mandibleinvolved the angle of the horizontal and vertical ramusof the mandible. Based on radiography and locationof the fractures it was decided to repair the fractureusing non-locking reconstruction plates at bothfracture sites.

Treatment and Discussion

The animal was initially stabilized with promptfluid therapy using dextrose 10% and Ringers lactatedsolution. Inj. Cefotaxim @ 20 mg/kg i/v was initiated.The surgery was scheduled after 2 days of initial

Surgical management of bilateral mandibular fractures in canine usingnon-locking reconstruction plates

K.S. Chaudhari, S.D. Tripathi, Aakash Vedpathak, D.U. Lokhande, G.S. Khandekar,Maneesh Tiwari, Tenzing Sherpa

Department of Veterinary Surgery and Radiology, Bombay Veterinary College.

presentation and the dog was admitted into the inhouse facility of the hospital. Prior to surgery the dogwas fasted for 12 hours. Inj. Triflupromazine @ 1 mg/kg was given as preanaesthetic and dog waspremedicated with Inj. Atropine sulphate @ 0.02 mg/kg. The oral cavity was flushed and cleaned with dilutebetadine solution and site was prepared aseptically.Induction of anaesthesia was done using 5%thiopentone sodium @ 12 mg/kg i/v and maintenancewas done using 5% Thiopentone administeredintravenously to effect. The fractured fragments wererepositioned into normal anatomical position so as toallow normal occlusion of the mandibular andmaxillary teeth. On the right side the plate was fixedin a horizontal orientation along the length of thehorizontal ramus using screws of 2.5 mm diameter.Drilling of the mandible was done using a manualhand drill and care was taken to avoid the roots of themandibular teeth. The fracture of the left mandiblewas visualized better by taking an incision on the angleof the mouth. The plate was fixed in an oblique mannerto involve both the horizontal and vertical rami foradequate support. The plates were covered withgingival tissue. The oral cavity was thoroughlycleaned with dilute povidone iodine solution. Inj.Meloxicam @ 0.2 mg/kg s/c was given for painmanagement. The dog recovered from anaesthesia

Fig. 1 : Placement of plate and screws intra operatively priorto closure of gingival tissue

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uneventfully. The muzzle of the dog was bandaged toavoid movement for 24 post surgery. Clinicalexamination 2 days post operatively showed normalocclusion of the jaw. The dog was able to drink liquidvoluntarily and was also able to vocalize normally.Solid food was withheld for 7 days postoperatively.On 8th day after surgery animal was able to eat solidfood. The case was discharged 10 days postoperatively. The case was examined 4 weeks and 8weeks post surgery. The dog showed no discomfortor complication. Mild halitosis was observed by theowner which was treated with cleaning of oral cavityand brushing of teeth twice a week.

Thus the fixation of miniplates was found to bea suitable technique for repair of mandibular fractureinvolving the horizontal body or the angle of themandible.

REFERENCES

Boudrieau, R.J. (2012). Maxillofacial fracture repairusing miniplates and screws. In: Oral andmaxillofacial surgery in dogs and cats: 293-308.

Boudrieau, R.J. and Kudisch, M. (1996). Miniplatefixation for repair of mandibular and maxillaryfractures in 15 dogs and 3 cats. Vet Surg; 25: 277–91.

Boudrieau, R.J., Tidwell, A.S. and Ullman, S.H.(1994). Correction of mandibular nonunion andmalocclusion by plate fixation and autogenous corticalbone grafts in two dogs. J Am Vet Med Assoc; 204:744–50.

Freitag, V. and Landau, H. (1996). Healing of dentateor edentulous mandibular fractures treated with rigidor semirigid plate fixation—an experimental study indogs. J Cranio maxillo facial Surg. 24 (2): 83-87.

Fig. 2 : VD radiograph of skull showing proper alignmentof fracture fragments post operatively.

Fig. 3 : Lateral radiograph of skull showing proper alignmentof fracture fragments post operatively.

the journal of bombay veterinary college · 2019. 3. 22. · sangram chavan, d.s. deshmukh, amol...

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