biochemical and histological studies of hamster (mesocricetus...

Middle East Journal of Applied Sciences ISSN 2077-4613

Volume : 08 | Issue :01 |Jan.-Mar.| 2018 Pages: 157-170

Corresponding Author: Abdel Rahman Bashtar, Zoology Department, Faculty of Science, Cairo University, Giza, Egypt.

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Biochemical and Histological studies of hamster (Mesocricetus auratus) infected with Schistosoma haematobium cercariae

Abdel Rahman Bashtar1, Fayez A. Bakry2, Hoda M. shoman3, Nadia F. Ibrahim3

and Entsar R. Farag3

1Zoology Department, Faculty of Science, Cairo University, Giza, Egypt. 2Medical malacology, Theodor Bilharz Research Institute (TBRI), Giza, Egypt. 3Zoology Department, Faculty of Science (for girls), Al-Azhar University, Cairo, Egypt and Biology Department, The University College of Khafji, Hafr albatin University, Saudi Arabia

Received: 18 Dec. 2017 / Accepted: 11 Feb. 2018 / Publication date: 25 Feb. 2018 ABSTRACT

The present work evaluated the effect of infection with Schistosoma haematobium cercariae on biochemical parasitological parameters and histological structure of serum and liver of hamster. The present results showed a significant decrease in the activity of transaminase enzymes (Aspartate amino transferase (AST) and Alanine amino transferase) in Serum and liver tissue of infected hamsters than that in the control hamsters. Electrophoretic analysis showed a remarkable deviation in the serum total protein of hamsters infected with S. haematobium(39.32%) in comparison with control non infected group. In addition, there was a remarkable deviation (40.44%) in total protein of liver homogenate of hamsters infected with S. haematobium in comparison with the control non infected group. There are a viable Bilharzial egg deposition, with related granulomas of chronic inflammatory cellular collection having a portal location in the liver of hamster infected with S. haematobium cercariae. On conclusion, S. haematobium causes serious histopathological changes on liver of hamsters as well as protein profile both of serum and liver homogenate.

Key words: Male hamsters, Schistosoma haematobium, cercariae. Electrophoretic, Histological and

Biochemical studies, analysis.

Introduction

Schistosomiais is prevalent in tropical and sub-tropical areas, especially in poor communities without access to safe drinking water and adequate sanitation (WHO, 2010). Data published by the World Health Organization (WHO) today show that almost 90 million individuals were treated for schistosomiasis in 2016, including 70.9 million school-age children and 18.3 million adults. (WHO, 2017). Of the 207 million people with schistosomiasis, 85% live in Africa (WHO, 2009). Water resource schemes for power generation and irrigation have resulted in a tremendous increase in the transmission and out breaks of schistosomiasis in several African countries (Sarkinfade et al., 2009]. Schistosomes are blood trematodes that are perfectly adapted to living in their intravascular habitat and to achieve this they have developed mechanisms enabling them to evade the immune and hemostatic responses of the host and to regulate endothelial cell function to favor their own survival (Eduardo, et. al., 2014) .Schistosomiasis, also known as bilharziasis, results from long-lived infection by multicellular intravascular parasites of one of five trematode species Schistosoma japonicum, Schistosoma mansoni, S. haematobium, Schistosoma intercalatum, or Schistosoma mekongi. Electrophoresis is the ability to separate a polypeptide of interest and to have an indication of its molecular size .It is very important in any study involving mixtures of proteins. The most relatively simple and powerful technique involves sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). In this method, separation of proteins based on their molecular size where, the SDS –protein complexes are sieved through a polyacrylamide gel matrix. The combination of SDS and sieving properties in the molecular sized pores of the gel matrix, leads to an exceedingly high resolution of separation that is unattainable with any other separation method based upon protein size (Rollinson and Simpson, 1987).Acute schistosomiasis has a significant impact on specific liver

Middle East J. Appl. Sci., 8(1): 157-170, 2018 ISSN 2077-4613

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functions and the alterations in specific protein isoforms and up regulation of unique proteins may be valuable as new markers of disease (Harvie et al., 2007).

Parasite transmission and the consequent risk of human infection are strongly linked to specific geographic locations, because the parasite goes through several developmental stages that must occur in fresh water, including a period of growth within particular species of intermediate host snails (King, 2009). Accumulation of S. haematobium eggs around the bladder and ureters leads to granuloma formation and fibrosis. In addition, calcification of dead eggs in the bladder wall results in rigidity of the bladder and subsequent increased pressure in ureters and kidneys. The bladder epithelium develops pseudopolyps, which can transform into squamous cell carcinoma in untreated patients (Knirsch, 2005). Spicher, et al.( 2004) mentioned that eggs of schistosome could either succeed in reaching the lumen of the organ (intestine or bladder) and leave the body with urine or feces or remain trapped in body tissues where they die and induce granuloma formation or even be transported in the blood flow toward other organs (including peritoneum) where they determine granulomas. Zampachova, et al.( 2006) mentioned that among the general histopathological changes, many lesions that correspond to an active chronic infection with a chiefly granulomatous reaction. At the same time, calcification, pseudotumorous polypoid formations, ulcerations, obliteration fibrous lesions, epithelial transformations such as hyperplasia, metaplasia and dysplasia were usually seen.

The present work aims to evaluate the effect of infection with S. haematobium cercariae on some biochemical and histological parameters in liver of hamster after infections.

Materials and Methods Experimental animals

Sixty six male hamsters were provided by lab/bred colony of similar age and weighing 100-120 g were selected for this study. Animals were kept in a controlled animal room at 24oС±2 and were allowed free access to diet and water during the study.

Cercariae

Schistosoma haematobium cercariae were obtained from Schistosome Biological Supply Center (SBSC) at Theodor Bilharz Research Institute (TBRI), Imbaba, Giza, Egypt. Infectivity of S. haematobium cercariae to hamster Body immersion technique for hamster:

Forty five hamsters were first immersed in warm water at 35 ˚C for 30 minutes to stimulate them to defecate and urinate, and to prepare their skin for cercarial penetration (Watson et. al., 1948). Glass bottle containers with capacious neck, 15 cm high, and of adequate size were filled with dechlorinated water at 30˚C to a height of 3-4 cm from the bottom (De witt, 1965). A sufficient amount of cercarial suspension containing 120 cercariae was withdrawn by means of a 5 ml glass pipette provided with an upper syringe attachment and introduced into each container. Hamsters were left in the bottles for one hour to ensure penetration of the maximum number of cercariae. Animals were then transferred carefully to their prepared cages. They were kept on a well-balanced and adequately nourishing diet to avoid malnutrition that would inhibit egg production (warren, 1970).Hamsters were sacrificed by decapitation. No resort to general anaesthesia or heparin was attempted because of the hepatic shift they produce. (Duvall and Dewitt, 1967).

Sacrifice and perfusion of hamster to detect adult worms

Twelve hamsters were first skinned out and their bodies were then washed with tap water to remove adherent hairs. Forty-five days after exposure to cercariae, hamsters in each group were sacrificed individually and dissected. The worm load in each hamster was carried out by perfusion


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(hepatic and intestinal) according to the method of (Smithers and Terry, 1965). The mean number of worms/mouse was considered as an indicator on the infectivity of cercariae in each experiment (Smithers and Terry, 1965).

Egg Developmental Stages Percentage (Oogram Pattern)

The oogram (percent eggs developmental stages) was studied by the method of Pelligrino and Goncalves, (1965). Three fragment (each 1 cm long, were cut off from the small intestine of the same 12 infected hamsters) used in the study of worm burden. Each fragment was examined microscopically and one hundred eggs were counted in each fragment. The different developmental stages of ova are investigated as follows:-The immature stage contains abnormal miracidia; the mature stage contains a fully developed miracidia. In addition, the dead ova were counted and recorded. The number of eggs/gram tissue (liver and intestine) of infected hamster was determined (Kamel, et al., 1977).

Biochemical studies Preparation of serum of hamster:

2-3 ml blood sample was taken immediately after scarification of hamsters in centrifuge tubes. The blood samples were centrifuged at 3000 Rpm for 10 min at + 4C˚.The obtained serum was preserved at -20 C˚ until used for biochemical studies.

Preservation of liver for biochemical studies

From each experimental animals a portion of 2.5 g was taken covered with aluminum foil and stored at -20oc until used for biochemical assays (enzyme determination and electrophoresis).The enzyme assay of liver functions was determined on portions weighing 0.34g from each liver, after homogenization in 2.5 ml of the specific solution to give 10% concentration and then used for determination. (Lowry et al., 1952).

Enzyme determination by spectrophotometer:

All physiological parameters determined in this study were determined spectrophometrically, using reagent kits purchased from Bio-Merieux Company, France. Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) activities were determined according to the method of Reitman and Frankel, (1957). Acid and alkaline phosphatase (ALP) activities were determined according to King and King, (1954).

Electrophoretic analysis:

The protein profiles were analyzed by SDS-PAGE according to the procedure of Boswell et al. (1987). Electrophoresis on SDS-PAGE a lab gel was fixed in 505 ml methanol hydrated in distilled water and stained with Commassie for 15 min. The gel was washed with distilled water and soaked in the developer until bands appeared. High and low molecular weight standards (marker 116= beta gluctosidase,marker 97.4 =phosphorylase B ,marker 66.2=bovine serum albumin, marker 37.6 = carbonic anhydrase and marker 28.2= triose phosphate isomerase) were electrophoresed on the same gel to calculate the relative molecular weights of the examined antigens. The gel was dried at room temperature, photographed using Kodak Tri-X-pan films and the molecular weights and protein intensity were analyzed by using Gel Docu Advanced software program.

Histological studies:

Three groups (each of six hamsters) were infected with S.haematobium and six hamsters were used as a control group. After sacrification of animals, part of the liver from each animal was


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removed, fixed in Bruin's fluid for five hrs, then transferred to 70% alcohol. Further procedures included dehydration in 100% alcohol, clearing in xylol and paraffin embedding were followed. Four sections (5 microns in thickness) were taken from each liver sample, each section being at a distance of at least 500um from the proceeding one. Sections were stained with haematoxylin and eosin and were examined under a polarized light microscope.

Statistical analysis

Electrophoretic analysis was carried out using dendrogramatic analysis of the electrophoretic profile of hamster liver and serum homogenates (Dice, 1945). The obtained data were analyzed by the student"s t-test for comparing means of the experimental and control groups (Spiegel, 1981).

Results Infection of hamsters with Schistosoma haematobium cercariae

The infectivity of S. haematobium cercariae, their development to adult worms within hamsters, the number of ova per gram tissue and oogrampatterns were estimated.

Worm load in infected hamsters

The results showed that the mean number of worm pairs worms (44.6±5.18) was greater (p<0.001) than that of both male and female worms (Fig. 1). The mean number of male (12.1±2.23) worms was greater than those of female worms (3.1±0.99).

Mea

n n

um

ber

of w

orm

s/ha

mst

er

Fig.1: Worm load of Schistosoma haematobium in hamsters infected with cercariae

Developmental stages of ova (oogram pattern).

The mean number of mature stages of schistosome ova (13276.8/g tissue) is higher in the intestine of hamster than the immature stages (3688/g tissue) ova. Meanwhile, the mean number of dead ova being 1475.2 was lower in the intestine of hamster with highly significant difference than mature and immature ova (p<0.001) (Fig.2).


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% D

evel

opm

enta

l st

ages

of

ova

Fig. 2: Developmental stages of ova (oogram) in the small intestine of hamsters infected with

Schistosoma haematobium cercaria

Effect of infection with Schistosoma haematobium cercariae on biochemical parameters of serum and liver of hamster. Liver function enzymes

The results revealed that transaminases enzymes (liver function) were significantly reduced in Serum and liver tissue of infected hamster with S. haematobium (Fig. 3) than control normal hamster. The concentration of transaminase enzymes (Aspartate amino transferase (AST) and Alanine amino transferase) in Serum and liver tissue of infected hamster showed more significant decrease than in the control hamster, The percentage reduction was 29.5% for AST and 23.8% for ALT in serum and 15.89% for AST, and 11.2% for ALT in liver tissue of hamster.

The present results (Fig. 3) indicated that there were significant elevations in the level of acid phosphatase, 40. 46% in serum, and 40.46% in liver tissue of hamster infected with S. haematobium. The increase of alkaline phosphatase was 50.61% in serum and 52.2% in liver tissue of infected hamster.

Fig. 3: Percentage of activity of AST and ALT enzymes in serum and liver homogenate of hamster infected with Schistosoma haematobium

60

50 40 30

20 10

0 -10

-20 -30

AST ALT ALKP ACP

Enzyme

liver Serum

Per

centa

ge o

f activ

ity o

f enzy

me


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Electrophoretic analysis

The pattern of protein profile identified by SDS-PAGE electrophoresis for serum of hamster infected with S. haematobium was shown in Fig. 4 a. Fig. (4 b) showed that the protein profiles of serum of normal hamster (lane1) and serum of infected hamster 2 are composed of 11 protein bands. This profile was reduced to 9 bands in the serum of infected hamster 1. The molecular weights of these bands for serum of normal hamster ranged from 120.91 to 31.563 KDa. Those for infected hamster 1 ranged from 126.02 to 31.563KDa and serum of infected hamster 2 ranged from 120.91 to 31.563 KDa.

Data showed the appearance of bands in infected groups and disappearance of others in comparison with control group. The disappearing 4 bands are 38.271, 78.777, 105.54 and 109.58KDa, while 5 bands appeared in serum infected hamster 1and 2 .e.g. 45.269, 71.964, 87.443, 118.89and 126.02KDa for serum infected hamster 1 and 36.188, 50.334, 73.738, 84.747 and 98.797 for serum infected hamster 2 (Fig.4 a & b).

Also, two shared bands (31.563 and 34.638 KDa ) appeared in protein profile of control and serum infected hamster 1, while three shared bands (31.563 , 34.638 and 120.91KDa) appeared in protein profile of control and serum of infected hamster 2. The shared bands seemed not to be affected by infection in spite of the variation shown in their amount of protein. The present results showed qualitative and quantitative differences in protein expression and banding pattern between infected and control hamster.

The present result in Fig. (4 b) displayed dendrogramatic analysis of the electrophoretic profile of hamster serum showed a remarkable deviation (39.32%) in hamsters infected with Schistosoma haematobium in comparison with control non infected group. The pattern of protein profile identified by SDS-PAGE electrophoresis for liver homogenate of hamster infected with S. haematobium was shown in fig. (5). Protein profiles of liver of normal hamster (lane1) and liver infected hamster 1(lane 2) are composed of 10 protein bands. These profiles were reduced to 9 bands for liver of infected hamster 2 (fig. 5a). The molecular weights of these bands for liver of normal hamster ranged from 120.44 to 30.047 KDa. Those for liver infected hamster 1 ranged from 120.44 to 31.701 KDa and liver infected hamster 2 ranged from 123.76 to 33.889 KDa.

The present data (Fig.5a) showed the appearance of bands in infected groups and disappearance of others in comparison with the control group. The disappearing 5 bands are 30.047, 38.271, 78.777, 105.54 and 109.58 KDa, while 5 bands appeared in liver infected hamsters 1and 2 .e.g. 31.701, 34.224 , 43.312, 85.935and 111.44 KDa for liver infected hamster 1 and 35.135, 50.781, 69.263, 108.73 and 123.76 for liver infected hamster 2.

One shared band (120.44KDa ) appeared in protein profile of control and liver infected hamster 1 while three shared bands (33.047, 90.854 and 114.51 KDa) appear in protein profile of control and liver infected hamster 2. The shared bands seemed not to be affected by infection in spite of the variation shown in their amount of protein. The present results showed qualitative and quantitative differences in protein expression and banding pattern between infected and control hamsters.

The present result in (Fig. 5 b) displayed dendrogramatic analysis of the electrophoretic profile of hamster liver homogenate. The analysis showed a remarkable deviation (40.44%) in the liver homogenate of hamster infected with S.haematobium in comparison with control non infected group.


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Effect of infection with Schistosoma haematobium cercariae on histological structure of tissues of hamster

The present results showed that histologically, the liver of hamster is formed of homogeneous mass of parenchymal cells arranged in hepatic lobules, distinguished by their central vein and separated by poorly developed interlobular septae. The hepatic lobules are composed of irregular branched and interconnected hepatic strands that anastomose to form a network enclosing a system of tortuous blood sinusoids. The hepatic cells are large in size, polygonal in shape and possess well-defined cell boundaries and a large central vein. Liver of normal hamster (Pl.1Figs.a&b) showed preserved normal hepatic lobule with radial arrangement of hepatocytes around the central vein. No fibrosis, cirrhosis, dysplasia or neoplasia and or degeneration was recorded of hepatocytes.

On the contrary, the results showed viable bilharzial eggs deposition, with related granulomas of a chronic inflamatory cellular collection having a portal location in the liver of hamster infected with S. haematobium cercariae (Pl.1. Figs. c&d). Viable eggs appeared oval in longitudinal section or round showing the miracidial content in cross- section (Pl.2 Fig. a). It is rimmed by chronic granulomatous inflammatory exudates of lymphocytes, plasma cells and foamy histiocytes.


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PI. 1: a, b): Section in Iiver of normaI hamster showing preserved normaI hepatic structure with radiaI arrangement of hepatocytes around the centraI vein and separated by bIood sinusoids. H=hepatocytes, CV=centraI vein, BS= bIood sinusoids. C & d): S. in Iiver of infected hamster with Schistosoma haematobium cercariae showing viabIe BiIharziaI eggs deposition, with reIated granuIomas of a chronic infIamatory ceIIuIar coIIection having a portaI Iocation. G= granuIoma, BE= BiIharziaI egg, lC= infIammatory ceIIs, NS = necrotic spaces. (c. H&E x100 ) (d. H&E x300 ).

Pl. 2: a): Section of a viablel bilharzial egg infestation with a retraction artifact, shinny shell & miracideal content surrounded by a large number of inflammatory cells. DH=degenerated hepatocytes, NS=necrotic spaces, BE= bilharzial egg, IC= inflammatory cell, MC= miracidial content, TS= terminal spine b): S. in liver of hamster showing a multinucleated histiocytic inflammatory giant cell with cytoplasmic engulfed foreign bilharzial pigment. BP= Bilharzial pigment, MC= multinucleated cell (H & E x 400). C): An inflammatory giant cell, engulfing a recent viable bilharzial egg. BE= Bilharzial egg, MC=multinucleate cell, NS= necrotic spaces (H&E x400).


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PI. 3: a): Section in Iiver of hamster showing a cross section of S. hematobium worm with surrounding 2ry induced peri−portaI fibrosis. F=fibrocytes, W=worm, NS=necrotic spaces (H&E x 400). b) S. in Iiver of hamster showing diIated vascuIar spaces (centraI vein) of portaI circuIation, CV= centraI vein, no cirrhosis, dyspIasia or neopIasia (H&E x200).

PI. 4: a & b): Diffuse, tri−zonaI mixed both micro & macro−vesicuIar steatosis “fatty change”, but no cirrhosis, dyspIasia or neopIasia, S=steatosis, CV= centraI vein (a. H & E x 200 & b. H & Ex 100). C): Low power of peri−portaI fibrosis with an associated oId granuIoma & pigment deposits, no cirrhosis, dyspIasia or neopIasia, BP= BiIharziaI pigment (H&E x200).

Infection of hamster with S.haematobium cercariae caused multinucleated histolytic

inflammatory giant cell with cytoplasmic engulfed foreign bilharzial pigment and a recent viable bilharzial egg (Pl.2 Fig. b&c).The present results in (Pl.3: a) showed a cross section of S. hematobium worm with surrounding 2ry induced peri-portal fibrosis Pl.3:b, a dilated vascular spaces (central vein) of portal circulation, due to backpressure 2ry to peri-portal fibrous obliteration but no cirrhosis, dysplasia or neoplasia. Infection of hamster with S. haematobium cercariae caused liver fatty changes (Pl.4:a&b) but no cirrhosis, dysplasia or neoplasia. Pl.4:c, showed peri-portal fibrosis with an associated old granuloma & pigment deposits.

Discussion

Infectivity of S. haematobium cercariae to hamster showed that the mean number of paired worms was greater than those of both male and female single worms. This agrees with the findings of Ritchie, et al. (1974) who recorded that infectivity of S. mansoni cercariae was inhibited after treatment with 100 ppm of bis tri-nbutyltin oxide for 5 min. The same findings were observed by


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Viyanant, et al. (1982) who used sublethal concentrations of copper sulphate and tributyltin fluoride, while Gawish, (1997) who used sublethal concentrations of niclosamide.

In the present work, the mean number of mature stages of Schistosome ova was higher in intestine of hamster than immature stages. Meanwhile, the number of dead ova was lower in intestine of hamster with highly significant difference than mature and immature ova.These results agree with the observations of El-Ghayeb, (1970) who stated that exposure of S. mansoni cercariae to 0.25 ppm of copper sulphate for 15-60 min markedly decreased the number of recovered worms per infected mouse and the number of ova/g tissue of liver. The reduction in the number of ova was explained as the few specimens of cercariae that survived after the treatment with molluscicides could infect the exposed mice and developed to adult worms laid low number of ova (WHO, 1963). This may be due to the disturbance in their physiological activities as molluscicide affects the respiratory enzymes, which are essential factors in the physiological processes of cercariae and adult worms. Also, in this work, the number of mature ova in the experimental groups was lower significantly in groups of mice infected with the Schistosome cercariae shed from Biomphalaria snails exposed to Earth Tec.

The present results showed that viable bilharzial eggs deposition, with related granulomas of a chronic inflamatory cellular collection having a portal location in the liver of hamster infected with S. haematobium cercariae. Eggs appeared oval in longitudinal section or rounded in shape on cross-section. This agree with Rollino et al. (2008) who claimed that egg production of S. haematobium starts 4-6 weeks after infection and continues for the life of the worm. Eggs pass from the lumen of blood vessels into the adjacent tissues. Spicher et al. (2004) mentioned that eggs of schistosome can either succeed in reaching the lumen of the organ (intestine) and leave the body with urine or feces or remain trapped in the body tissues, where they die and induce granuloma formation or even be transported in the blood flow toward the other organs(including peritoneum) where they determine granulomas.

The data obtained showed that Infection of hamster with S.haematobium cercariae caused a multinucleated histolytic inflammatory giant cell with cytoplasmic engulfed foreign bilharzial pigment. Also, shows an inflammatory giant cell, engulfing a recent viable bilharzial egg. These observations agree with Rollino et al. (2008) who observed that in chronic schistosomiasis, tissue injury is mediated by egg-induced granulomas and there is a subsequent appearance of fibrosis. Both Th1 and Th2 cytokines are involved in the granulomatous response. But granuloma growth and maintenance are largely associated with Th2 cytokines, down -regulated by interleukin 12.Enzymes and antigens released from eggs activate sensitization of the host lymphocytes, which migrate to areas of egg deposition and recruit other cell types, such as macrophages, eosinophils, and fibroblasts. The size of these granulomas and the resulting fibrosis lead to most of the chronic fibro-obstructive lesions in the schistosomiasis. Also, Michael and Anthony, (1998) mentioned that in granulomatous reactions, the eggs induction leads to fibrosis. This in turn may lead to portal hypertension or urinogenital dysfunction, depending on the parasite species. The disease symptoms are therefore attributable to immunopathology. The granuloma is a T- cell dependent, delayed –type hypersensitivity reaction comparable to that seen in other chronic granulomatous conditions, except that eosinophils as well as lymphocytes and macrophages are abundant in the lesions.

Infection of hamster with S. haematobium cercariae caused peri-portal fibrosis with an associated old granuloma & pigment deposits and dilated vascular spaces (central vein) of portal circulation, due to backpressure 2ry to peri-portal fibrous obliteration. This agrees with Chiti et al. (1999) who reported large numbers of schistosome worm pairs and eggs in the hepatic veins.

Furthermore, no cirrhosis, dysplasia or neoplasia were observed. This may be due to fibrosis, which leads to elevation of the portal pressure that causes dilation of the portal veins (Morgan et al., 2005). Moreover, Zampachova (2006) announced that among the general histopathological changes seen were lesions that correspond to an active chronic infection with a chiefly granulomatous reaction which mostly followed by calcification, pseudotumorous polypoid formations, ulcerations, obliteration fibrous lesions and epithelial transformation such as hyperplasia, metaplasia and dysplasia follow later. On the other hands, Neves et al. (2006) mentioned that Schistosoma mansoni infection with high-fat diet promotes intensive quantitative changes in hepatic structure, contributing to an increase of hepatic regeneration. They observed that only liver of fatty mice exhibited some degree of steatosis.


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Transaminases [aspartate aminotransferase (AST) and alanine aminotransferase (ALT)] represent an important link between carbohydrate and amino acid metabolic pathways (Christic and Michelson, 1975). Also, these enzymes are considered as good sensitive tools for detection of any variations in the physiological process of the living organisms (Tolba et al., 1997).

Concerning AST and ALT enzyme activities, significant reduction was observed in the liver tissue of infected hamsters. The decrease observed in AST and ALT attributed to the hepatocellular damage resulted from egg deposition where the transaminases level showed an intimate relationship to cell necrosis and / or increased cell membrane permeability that led to the discharge of the enzyme to the blood stream (El-Aasar et al., 1989 and El-Shazly et al., 2001). The decrease in the transaminases level providing additional support for the side effect of the S. mansoni infection on mitochondria of the hepatic cells as it is the subcellular localization of transaminases (Mansour et. al., 1982).

In the present study, Acid and alkaline phosphatases (ALP) show significant elevation in infected hamsters. Higher levels of acid and alkaline phosphatases (ALP) in tissues was observed by El-Aasar, et al. (1989) and Abdel-Rahman et al. (1993). They attributed it to the irritation of liver cells by toxins or metabolic products of growing schistosomoules of adult worms and eggs or due to the increased loss of the intracellular enzymes by diffusion through the cell membrane that appear to act as a stimulus to the synthesis of more enzyme.

SDS-PAGE of whole cell proteins is a useful technique for identification of isolates complex. The present results indicated that infection of hamster with S. haematobium had qualitative and quantitative effect on the protein patterns of serum and liver tissues of infected hamster. The electrophoretic pattern of the native proteins revealed difference in the number and molecular weight of protein bands compared to the control hamsters. These differences indicated that infection of hamster with S. haematobium caused intensive effects that induced fractionation of the native protein. This agree with Bakry et al. (2007), who observed two characteristic bands (140.82KD&14.42KD) in the electrophoretic patterns of tissue proteins from B. truncatus snails infected with E.recurvatum at intervals of two and four weeks post infection, but there is only one band (100.9KD) characteristic for B. truncatus snails infected with S. haematobium snails.Also, EL-Dafrawy, et. al., (2006) showed qualitative and quantitative differences in the protein expression and banding patterns between non-infected and infected B.alexandrina and B.truncatus snails. However, in Biomphalaria snails, the molecular weight of the plasma proteins found to be ranged between 10 and 450 KDa, this may be due to, all strains show protein patterns, although minor inter-and intrastrain differences occur. Accordingly, the fractionation of native proteins into bands different from that of the control may be attributed similar to changes occurred in DNA of the treated snails (El- Sayed, 2006). The data obtained showed that Infection of hamster with S. haematobium cercariae caused a multinucleated histolytic inflammatory giant cell with cytoplasmic engulfed foreign bilharzial pigment. Also, shows an inflammatory giant cell, engulfing a recent viable bilharzial egg. These observations agree with Rollino et al. (2008) who observed that in chronic schistosomiasis, tissue injury is mediated by egg-induced granulomas and there is a subsequent appearance of fibrosis. Both Th1 and Th2 cytokines are involved in the granulomatous response. But granuloma growth and maintenance are largely associated with Th2 cytokines, down -regulated by interleukin 12.Enzymes and antigens released from eggs activate sensitization of the host lymphocytes, which migrate to areas of egg deposition and recruit other cell types, such as macrophages, eosinophils, and fibroblasts. The size of these granulomas and the resulting fibrosis lead to most of the chronic fibro-obstructive lesions in the schistosomiasis. Also, Michael and Anthony, (1998) mentioned that in granulomatous reactions, the eggs induction leads to fibrosis. This in turn may lead to portal hypertension or urinogenital dysfunction, depending on the parasite species. The disease symptoms are therefore attributable to immunopathology. The granuloma is a T- cell dependent, delayed –type hypersensitivity reaction comparable to that seen in other chronic granulomatous conditions, except that eosinophils as well as lymphocytes and macrophages are abundant in the lesions.

Infection of hamster with S.haematobium cercariae caused peri-portal fibrosis with an associated old granuloma & pigment deposits and dilated vascular spaces (central vein) of portal circulation, due to backpressure 2ry to peri-portal fibrous obliteration. This agrees with Chiti et al. (1999) who reported large numbers of schistosome worm pairs and eggs in the hepatic veins.


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Furthermore, no cirrhosis, dysplasia or neoplasia were observed. This may be due to fibrosis, which leads to elevation of the portal pressure that causes dilation of the portal veins (Morgan, et. al., 2005). Moreover, Zampachova (2006) announced that among the general histopathological changes seen were lesions that correspond to an active chronic infection with a chiefly granulomatous reaction which mostly followed by calcification, pseudotumorous polypoid formations, ulcerations, obliteration fibrous lesions and epithelial transformation such as hyperplasia,metaplasia and dysplasia follow later. On the other hands, Neves et al. (2006) mentioned that Schistosoma mansoni infection with high-fat diet promotes intensive quantitative changes in hepatic structure, contributing to an increase of hepatic regeneration. They observed that only liver of fatty mice exhibited some degree of steatosis.

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