USE OF ORGANIC ACIDS AS AN ALTERNATIVE TO

ANTIBIOTICS ON GROWTH, MEAT YIELD AND

ECONOMIC FEASIBILITY IN REARING SONALI CHICKS

A THESIS

By

MD. ISMAIL HAQUE

FARHAD NOMAN

Examination Roll No. 10AHPSJD- 04M Registration No: 16874

Session: 1989-90

Semester: July- December, 2011

MASTER OF SCIENCE (M.S.)

IN

POULTRY SCIENCE

DEPARTMENT OF POULTRY SCIENCE

BANGLADESH AGRICULTURAL UNIVERSITY

MYMENSINGH

NOVEMBER, 2011

USE OF ORGANIC ACIDS AS AN ALTERNATIVE TO

ANTIBIOTICS ON GROWTH, MEAT YIELD AND

ECONOMIC FEASIBILITY IN REARING SONALI CHICKS

A THESIS

By

MD. ISMAIL HAQUE

A.K.M. FARHAD NOMAN

Examination Roll No. 10AHPSJD- 04M Registration No: 16874

Session: 1989-90

Semester: July- December, 2011

Submitted to the Department of Poultry Science

Bangladesh Agricultural University, Mymensingh

In Partial fulfillment of the requirements

For the degree of

MASTER OF SCIENCE (M. S.)

IN

POULTRY SCIENCE

DEPARTMENT OF POULTRY SCIENCE

BANGLADESH AGRICULTURAL UNIVERSITY

MYMENSINGH

NOVEMBER, 2011

USE OF ORGANIC ACIDS AS AN ALTERNATIVE TO

ANTIBIOTICS ON GROWTH, MEAT YIELD AND

ECONOMIC FEASIBILITY IN REARING SONALI CHICKS

A THESIS

By

MD. ISMAIL HAQUE

A.K.M. FARHAD NOMAN

Examination Roll No. 10AHPSJD- 04M Registration No: 16874

Session: 1989-90

Semester: July- December, 2011

Approved as to the style and content by

(Prof. Dr. M. A. R. Howlider)

Supervisor

( Dr. Fowzia Sultana) Co-supervisor

___________________________

Dr. Shubash Chandra Das Chairman

Examination committee

&

Head

Department of Poultry Science

Faculty of Animal Husbandry

Bangladesh Agricultural University

Mymensingh

NOVEMBER, 2011

ACKNOWLEDGEMENTS

First of all, the author would like to express his sincere gratitude to “Almighty Allahˮ, the

supreme ruler of the universe the greatest, the most gracious and most merciful Who has

created all in and outside of the world and enabled the author for the completion of the

research work and in the preparation this thesis.

The author wishes with pleasure to express his sincere grateful, heartfelt respect, kind

regards, profound sense of appreciation and indebtedness to his teacher and Supervisor

Professor Dr. M. A. R. Howlider, Professor, Department of Poultry Science, BAU

Mymensingh for his scholastic guidance, untiring assistance, responsible suppuration,

constructive criticism, precious suggestions, continuous encouragement and affectionate

feelings throughout the course of the research work and in the preparation of this

manuscript.

The author also desires and feel proud to express his profound appreciation, veritable

gratefulness and heartfelt thanks to his Co-supervisor, Dr. Fowzia Sultana, Associate

Professor, Department of Poultry Science, BAU Mymensingh for her valuable guidance,

magnanimous help and helpful advice and constant creative instructions in the way to

accomplish this thesis.

The author would like to extend gratefulness, sincere appreciation and respect to

honourable teacher Dr. Md. Shawkat Ali, Associate Professor, Department of Poultry

Science, BAU Mymensingh for valuable suggestions and spending his valuable time for

data analysis.

The author desire to express his best regards and immense indebtedness to his respected

teachers Professor Dr. S. D. Chowdhury, Professor Dr. Md. Ashraf Ali, Professor Dr.

Shafiuddin (Rtd.), Professor Dr. S. M. Bulbul (Rtd.), Associate Professor Dr. Shubash

Chabdra Das and Associate Professor Dr. Bazlur Rahman Mollah, Department of Poultry

Science, BAU Mymensingh for their generosity and affectionate feelings and constant

inspiration at the various stages of academic and in this research work.

The author gratefully admires to his classmates, friends and specially his younger brother

Md. Zohurul Islam who arranged all facilities in carrying out this research in his owned

poultry farm located at Upazilla-Godagari, District- Rajshahi.

The author would like to express his cordial gratefulness to the authority of National

Agricultural Technology Project, Department of Livestock Services, Bangladesh for giving

all types of financial support in carrying out this research work.

Finally the author express his indebtedness to his parents for their heartfelt prayer,

affections, supports, sacrifice, inspiration, encouragement and continuous blessings in the

long process of building my academic career which can never be filled.

The Author

USE OF ORGANIC ACIDS AS AN ALTERNATIVE TO

ANTIBIOTICS ON GROWTH, MEAT YIELD AND ECONOMIC

FEASIBILITY IN REARING SONALI CHICKS

M.S. in Poultry Science

Examination Roll No.: 10AHPSJD-04M

Registration No.: 16874

Session: 1989-90

Semester: July-December 2010

ABSTRACT The present study was carried out to compare the effects of using organic acids and

antibiotics on the growth, meat yield and economic feasibility in rearing popular dual purpose

sonali (RIR × Fayomi) chicks seeking an alternative to antibiotic. A total of 240 straight run

sonali chicks were used. The chicks were fed on antibiotics (A); 0%, 0.01% ciprofloxacin

(C), 0.01% enrofloxacin (E) and 0.05% doxycycline (D) with or without Hameco-pH (organic

acid mixture as per manufacturer`s recommendation. The dose of Hameco-pH (H)

was 0.1%.

A and H were supplied with drinking water. The basal diet fed ad libitum contained 2950 kcal

ME/kg, 20% crude protein, 4.5% fibre, 6% crude fat, 1% calcium, 0.45% available

phosphorus, 0.48% methionine and 1.1% lysine up to 56 days of age. It was found that the

live weight, feed intake and feed conversion (there was an exception only at 7 days of age) of

sonali chicks at different ages were not influenced (p>0.05) by A, H or their combinations. At

56 days of age, recorded live weight differed (p<0.01) between sexes. Males had 25.87%

higher live weight than that of their female counterparts. The results represented highest

dressing yield (p<0.05) was attained on C and control groups than that on other 2 antibiotics;

E and D. Dressing yield was not altered (p>0.05) by H supplementation. Total meat and

breast meat were highest (p<0.01) and similar (p>0.05) in groups fed on control and E than

that of C and D. So, addition of A in the diet was not necessary for improving total and breast

meat. H did not alter (p>0.05) total and breast meat. Edible portion depleted by 0.91%, 2.01%

and 2.38% on E, C and D supplementation respectively than that on control. Positive

apparent difference was not observed for adding A in control diet. On the other hand, though

insignificant (p>0.05), but H had a positive trend to increase the dressing yield, edible portion

and drumstick meat of sonali chicks. Among the treatment; control, H, C, E, D, HC, HE and

HD, cost of production and mortality obtained was lowest on dietary H. Finally, it was

concluded that addition of antibiotics in diet of sonali chicks may not be useful to increase

growth and meat yield. On the other hand, organic acids had positive trend on growth

performance, meat yield and survivality of sonali chicks. The results obtained on growth

performance, meat yield and profitability revealed that organic acid may be a human health

friendly, safe and cost effective substitute of antibiotic.

LIST OF CONTENTS

Chapter Title Page

ACKNOWLEDGEMENTS i

ABSTRACT iii

LIST OF CONTENTS iv

LIST OF TABLES ix

LIST OF FIGURES x

APPENDICES xii ABBREVIATIONS AND SYMBOLS xiii

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 5

2.1. Antibiotics 5

2.1.1. Classification of antibiotics 6

2.1.1.1. Ciprofloxacin 7

2.1.1.2. Enrofloxacin 7

2.1.1.3. Doxycycline 8

2.1.2. Mode of action of antibiotics 8

2.1.3. Harmful effect of antibiotics 9

2.1.4. Human health hazard of antibiotic growth

Promoter

10

2.1.5. Antibiotics as growth and its effects on

production performance and meat yield of

poultry

11

2.2. Organic acids 13

2.2.1. Organic acids in poultry diet 14

2.2.2. Mode of action of organic acids 15

2.2.3. Antibacterial effect of organic acids 17

LIST OF CONTENTS (CONTD.)

Chapter Title Page

2.2.4. Site of action of organic acids 18

2.2.5. Organic acids on growth performance 19

2.2.5.1. Organic acids and live weight 19

2.2.5.2. Organic acids and feed intake 20

2.2.5.3. Organic acids and feed conversion 21

2.2.5.4. Organic acids and survivality 22

2.2.5.5. Organic acids and meat yield 23

3 MATERIALS AND METHODS 25

3.1. Statement of research work 25

3.2. Venue of the experiment 25

3.3. Collection of experimental materials 25

3.4. Experimental chicks 26

3.5. Layout of the experiment 26

3.6. Chemical composition of basal diet 27

3.7. The technique used in providing antibiotics and

Hameco-pH in the drinking water

27

3.8. Management practice 28

3.8.1. Preparation of house 28

3.8.2. Litter 28

3.8.3. Light, temperature and ventilation 28

3.8.4. Feed and water management 28

3.8.5. Bio security 29

LIST OF CONTENTS (CONTD.)

Chapter Title Page

3.8.6. Vaccination 29

3.9. Record keeping and calculations 29

3.9.1. Live weight 29

3.9.2. Feed consumption and feed conversion 30

3.9.3. Production cost and profitability 30

3.10. Slaughtering and dressing of chicks 30

3.11. Statistical Analysis 30

4 RESULTS 38

4.1. Growth performance 38

4.1.1. Live weight 38

4.1.2. Feed intake 38

4.1.3. Feed conversion 38

4.1.4. Cost of production 43

4.2. Meat yield characteristics 43

4.2.1. Live weight 43

4.2.2. Dressing yield 43

4.2.3. Total meat yield 44

4.2.4. Breast meat 44

4.2.5. Dark meat 44

4.2.6. Thigh meat 44

4.2.7. Drumstick meat 44

4.2.8. Wing meat 45

4.2.9. Edible portion 45

LIST OF CONTENTS (CONTD.)

Chapter Title Page

4.2.10. Abdominal fat 45

4.2.11. Skin weight 45

4.2.12. Liver weight 45

4.2.13. Heart weight 46

4.2.14. Gizzard weight 46

4.2.15. Neck weight 46

4.2.16. Head weight 46

4.2.17. Giblet 46

4.2.18. Blood loss 47

4.2.19. Feather weight 47

4.2.20. Drumstick bone weight 47

4.2.21. Wing bone weight 47

4.2.22. Drumstick bone length 47

4.2.23. Thigh bone length 48

4.2.24. Wing bone length 48

5 DISCUSSION 54

5.1. Live weight 54

5.2. Feed intake 54

5.3. Feed conversion 54

5.4. Survivality 55

LIST OF CONTENTS (CONTD.)

Chapter Title Page

5.5. Meat yield characteristics 55

5.5.1. Dressing yield 55

5.5.2 Breast meat 56

5.5.3. Abdominal fat 56

5.5.4. Heart weight 56

5.5.5 Liver weight 56

5.5.6 Gizzard weight 57

5.6. Economic efficiency of production 57

6 SUMMARY AND CONCLUSION 58

REFERENCES 59

LIST OF TABLES

Table Title Page

1. List of organic acids and their properties 14

2. Chemical composition of Hameco-pH 25

3. Chemical composition of antibiotics 26

4. Layout showing the distribution of day-old straight

run Sonali chicks to the levels and replications of the

antibiotics and Hameco-pH

.

26

5. Chemical composition of the basal diet 27

6. Vaccination schedule followed for the chicks 29

7. Live weight of Sonali chicks fed on different dietary

antibiotics with or without Hameco-pH

at different

ages

39

7. (Contd.) Feed intake of Sonali chicks fed on antibiotic with or

without Hameco-pH

at different ages 40

7. (Contd.) Feed Conversion Ratio of Sonali chicks fed on different

dietary antibiotic with or without Hameco-pH

at different

ages

41

7. (Contd.) Production cost/kg live weight and mortality of sonali

chicks fed antibiotiss with or without Hameco-pH

at

different ages

42

8. Meat yield characteristics of Sonali chicks fed on

different dietary Antibiotics with or without Hameco-pH

(H)

49

LIST OF FIGURES

Figure Title Page

1. Showing day old sonali chicks 32

2. Hameco-pH and Antibiotics (Enrofloxaacin,

Ciprofloxacin and Doxycycline)

32

3. Application of medicines in the water 32

4. Sonali chicks housed in different pens 33

5. Measuring live weight of male sonali chick 33

6. Measuring live weight of female sonali chick

33

7. Slaughtering chicks by Halal method

34

8. Recorded blood loss 34

9. Recorded feather loss 34

10. Dissection of chick 34

LIST OF FIGURES (CONTD.)

Figure Title Page

11. coloured shank of sonali chicks

35

12. Head 35

13. Neck 35

14. Drumstick meat with bone

35

15. Thigh meat with bone 36

16. Wing meat with bone 36

17. Breast meat 36

18. Spleen 36

19. Heart 37

20. Gizzard 37

21. Liver 37

22. Back 37

LIST OF APPENDICES

Appendix Title Page

1. Meat yield characteristics of Sonali chicks fed on

different dietary antibiotics with or without Hameco-

pH

74

2. Recorded temperature and relative humidity during

the experimental period 78

ABBREVIATIONS AND SYMBOLS

Abbreviation : Full meanings

AM. : Ante meridian

ANOVA : Analysis of Variance

BAU : Bangladesh Agricultural University

BCRDV : Baby chick Ranikhet Disease Vaccine

cm : Centimeter

Cm2

: Square centimeter

Contd.

: Continued

Cr : Chromium

CRD : Completely Randomized Design

DANMAP : Danish integrated Antimicrobial resistance

Monitoring And research Programme

DNA : Deoxy Ribose Nucleic Acid

D2O : Deuterium Oxide

et al. : And others

e.g. : For example

FA : Fumeric acid

FC : Feed conversion

FCR. : Feed conversion ratio

Fig. : Figure

g : Gram

GIT : Gastro intestinal tract

H+

: Cation of Hydrogen

i.e. : That is

kcal : Kilo-calorie

ABBREVIATIONS AND SYMBOLS (CONTD)

Abbreviation : Full meanings

kg : Kilogram

L : Liter

LSD : Least significant difference

MAFF : Ministry of Agriculture Fisheries and

Livestock

ME : Metabolizable energy

MIC : Minimum inhibitory concentrate

ME : Metabolizable energy

mg : Milligram.

ml : Milliliter

MRLs : Maximum residue limits

NH3 : Ammonium

No. : Number

NO2 : Nitrous oxide

NRC : National Research Council

NS : Non-significant

°C : Degree Celsius

p : Probability

ppm : Parts per million

PM : Post meridian

SED : Standard Error Difference

LIST OF ABBREVIATIONS (CONTD.)

Abbreviation : Full meanings

Sq : Square

Tk : Taka

µg : Microgram

% : Percentage

& : And

@ : At the rate of

< : Less than

> : Greater than

+ : Plus

/ : Per

* : 5% level of significant

** : 1% level of significant

CHAPTER I

INTRODUCTION

Bangladesh is considered as one of the most appropriate countries in the world for

rearing poultry. The poultry industry plays a crucial role in economic growth and

simultaneously, creates numerous employment opportunities (Shamsuddoha and

Sohel, 2003). Regardless of religion and age almost all people are fond of chicken

meat. People of any age can take poultry meat without hesitation for less content of fat

compared to other meats. We have to increase the animal protein production to make

our people sound and healthy. Protein intake is recommended to be in the range of 0.8

to 1.6g/d per kg body weight for human (Anonymous, 1998) requires minimum

20.44kg protein per person (average 70kg body weight) per year. It indicates that it is

a crying need to increase the meat production according to the requirements. In

Bangladeshi food culture, people always try to find the indigenous (Desi) cock for its

tenderness and good taste. One of the reasons is that poultry meat is still compact to

heat, but in broiler meat some portions are separated from the bone, that‟s why this is

not suitable for making roast. Majority people like cockerels weighing about 650-

700g, so that they can economically make maximum four roasts of 120-130g.

Practically, it is seen in the market that for a festival or usual consumption people

have been buying most of the cockerels. The demand of cockerels is bigger than that

of production. Local chicks could not meet the demand of the people in an

overpopulated country where about 142 million people living in an area of 143,999

square kilometers (Bangladesh Bureau of Statistics, March, 2011). For the necessity of

time Fayomi and Sonali (Rhode Island Red × Fayomi) growing straight run chicks

have been taking their place beside the indigenous chicks for their adaptability and

acceptability under the climatic conditions of Bangladesh (Anisuzzaman and Wahid,

1988). Moreover, they have tenderness and good taste as liked as Desi chicks. On

another observation, sonali crossbred stated as more suitable chicks for meat and egg

production to rear in rural areas for higher adaptability and disease resistance (Ali,

1989). Crossbred progenies were superior to purebred in growth rate, meat quality,

body weight and feed conversion compared to that of respective purebreds (Dubrynia,

1958; Masic and Khalifah, 1965). Crossbred Sonali might have a higher growth rate,

viability and meat yield because a certain level of hybrid vigor could be expected as

their parents Rhode Island Red (RIR) and Fayomi are 2 different breeds. RIR is bigger

sized chicks than that of Fayomi. The breed that will show better performance could

be recommended besides the native cockerels to partially fulfill the demand of meat.

In northern region of Bangladesh, sonali is more popular for meat production.

Highlighted concern over antibiotic resistance, natural alternatives; probiotic (Fuller,

1989) and some organic acid (Chaveerach et al., 2004) or combination of them are

used in the diets assume to be their positive effect on health and growth of broilers.

There are several types of organic acids; citric acid, acetic acid, lactic acid, fumeric

acid, malic acid, ascorbic acid etc. and also their different combinations are used in

poultry diets (Callsen, 1999). They have a specific antibacterial effect at a low pH and

may help to reduce overall bacterial numbers or modify bacterial species distribution

in the gut and increase nutritive value to the diet and thus improved their health.

Health of the gut is one of the major factors governing the performance of poultry.

The economics of poultry production (Samik et al., 2007) and the profile of intestinal

microflora play an important role in gut health (Dhawale, 2005). Dietary organic acids

and their salts are able to inhibit growth of microorganisms in the food (Nursey,

1997). Consequently, preserve the microbial balance in the GI tract. In addition, by

modifying intestinal pH organic acids also improve the solubility of the feed

ingredients, digestion and absorption of the nutrients (Vogt et al., 1981; Patten and

Waldroup, 1988; Owings et al., 1990 and Skinner et al., 1991). These discrepancies

would be related to the source, the amount of organic acids used, location,

environment and the composition of diets (Jensen and Chang, 1976; Gama et al.,

2000). Antibiotic growth promoters and antibiotic resistance are closely related. The

increased concern on potential for antibiotic resistant strains of bacteria has compelled

the researchers to utilize the other non therapeutic alternatives like enzyme, probiotic,

prebiotic, herb, essential oil, immune stimulant and organic acid as feed additives in

poultry production. Organic acids are not antibiotics but, if used correctly along with

nutritional, managerial and bio security measures, they can prove powerful in

maintaining the health of the GI-tract of poultry, on improving their zootechnical

performances. For antimicrobial effect, organic acids result in inhibition of intestinal

bacteria leading to reduced bacterial competition with the host for available nutrients

and diminution in the level of toxic bacterial metabolites resulted of lessened bacterial

fermentation improved protein and energy digestibility. The increased villous height

in the small intestines induced by organic acids increases the absorptive intestinal

surface, facilitates the nutrient absorption and growth performance. They decrease the

pH value in different segments of GI tract conducive to growth of favorable bacteria

simultaneously, hampering the growth of pathogenic bacteria grow at relatively higher

pH. The acid anion has been shown to complex with calcium, phosphorus, magnesium

and zinc improved digestibility of these minerals. Reduced gastric pH following

organic acid feeding increased pepsin activity and the peptides arising from pepsin

proteolysis trigger the release of hormones, including gastrin and cholecystokinin

regulate digestion and absorption of proteins.

Maximum dependency or imitational use of antibiotics as growth promoter and as

therapeutic in poultry is one of the important issue. Because, using antibiotics to

establish a beneficial condition in the GI tract has became disadvantage as well. These

days, antibiotic resistance (Bates et al., 1994 and Roy et al., 2002) and entering the

chemical drugs to human food chain are seriously considered. The rate of increase the

resistance to antibiotics (Neu, 1994) is a major public health hazard throughout the

world (Bengmark, 1998). This absurd use of antibiotics is not only developing an

increasing in microbial resistance to antibiotics but also causes of hypersensitivity

reaction or even cancer by the presence of residues in meat (Menter, 2002). Poultry

farmers are indiscriminately using different antibiotics for getting increased

performance and survivability. For this reason, the poultry sector is continuously

searching new feed additives in order to improve performance and health of chicks

and to avoid the undesirable effect of antibiotics.

Some researchers are suggesting the use of organic acids as a cheaper and safe

alternative to antibiotics, but limited studies have been done in Bangladesh to compare

the effects of using organic acids and antibiotics on the growth, meat yield and

economic feasibility of rearing popular dual purposes sonali chicks.

Keeping above information in mind, present research was aimed with the following

objectives:

1. To compare growth and meat yield of sonali chicks on diet with

antibiotic and organic acid supplementation.

2. To assess whether organic acid could be an alternative to antibiotic in

rearing dual purpose chicks.

CHAPTER II

REVIEW OF LITERATURE

Prevention of diseases and enhancement of growth are critical factors in modern

poultry production. Keeping these thinking in mind, poultry farmers are

indiscriminately using different antibiotics. But continuous and unnecessary use of

antibiotics cause antibiotic resistance and residual existence (Waldroup et al., 2003) in

poultry products, is the major health concern now a day. A considerable effort has

been made by poultry scientists to produce safe meat from poultry through the use of

different additives in diet or drinking water. Consumers around the world have major

concerns on safe food. Organic acids are now being used as most effective and

alternate additive of antibiotics to poultry diet considering safety of products and

human health. The present review outlines the summarization of published

information to evaluate the effects of organic acids.

2.1 Antibiotics

The word “antibiotic” comes from the Greek anti meaning 'against' and bios meaning

'life'. Antibiotics are also known as antibacterials, and they are drugs used to treat

infections caused by bacteria. The term antibiotic, meaning "against life," was

introduced by the French bacteriologist Vuillemin as a descriptive name of the

phenomenon exhibited by these early antibacterial drugs (Calderon et al., 2007 and

Calderon et al., 1974). According to Taber‟s Medical Dictionary “Antibiotics are

natural or synthetic substances that destroy microorganisms or inhibit their growth.”

According to 1943 edition of Webster‟s Dictionary “a substance produced by or a

semi synthetic substance derived from a microorganism and able in dilute solution to

inhibit or kill another microorganism.” Antibiosis was first described in 1877 in

bacteria when Louis Pasteur and Robert Koch observed that an airborne bacillus could

inhibit the growth of Bacillus anthracis (Landsberg, 1949). The term "antibiotic" was

coined by Waksman in 1942 to describe any substance produced by a microorganism

that is antagonistic to the growth of other microorganisms in high dilution (Waksman,

1947)

2.1.1 Classification of antibiotics

Antibiotics are commonly classified based on:

1. Chemical structure/biosynthetic origin:

a. Natural

b. Semi-synthetic and

c. Synthetic

2. Their mechanism of action.

3. According to their biological effect on microorganisms:

Most antibacterial antibiotics target bacterial functions or growth processes

(Calderon et al., 2007). In this classification antibiotics are divided into two broad

groups:

a. Bactericidal (kill bacteria): these are 3 types:

i) Those target the bacterial cell wall; penicillins and cephalosporins,

ii) Those target the Cell membrane; polymixins,

iii) Those Interfere with essential bacterial enzymes; quinolones and

sulfonamides.

b. Bacteriostatic agents; slow down or stall bacterial:

Those that target protein synthesis, aminoglycosides, macrolides, and

tetracyclines, are usually bacteriostatic (Finberg et al., 2004)

4. Spectrum of activity: This categorization is based on their target specificity.

i) Narrow spectrum; antibiotics target specific types of bacteria; Gram

negative or Gram positive bacteria and

ii) Broad spectrum antibiotics, affect a wide range of bacteria.

Following a 40 year hiatus in discovering new classes of antibacterial compounds, 3

new classes of antibiotics have been brought into clinical use. These new antibiotics

are: Cyclic lipopeptides; daptomycin, Glycylcyclines; tigecycline and Oxazolidinones

linezolid (Cunha, 2009).

Although there are well over 100 antibiotics, the majority come from only a few types

of drugs. These are the main classes of antibiotics: Penicillins; penicillin and

amoxicillin, Cephalosporins; cephalexin, Macrolides; erythromycin, clarithromycin,

and azithromycin, Fluoroquinolones; ciprofloxacin, levofloxacin and Enrofloxacin,

Sulfonamides; co-trimoxazole and trimethoprim. Tetracyclines; tetracycline and

doxycycline and Aminoglycosides; gentamicin and tobramycin.

2.1.1.1 Ciprofloxacin

Ciprofloxacin; a new fluoroquinolone highly active against many diverse

microorganisms. At concentrations of <1 μg/ml active against most gram negative

bacteria; Enterobacteriaceae, Haemophilus, Neisseria, and other Pasteurellaceae,

Vibrionaceae, and various species of Pseudomonas and Acinetobacter. Most

staphylococci, including strains resistant to methicillin, are also susceptible to

ciprofloxacin. Streptococci are not highly susceptible to ciprofloxacin, and obligate

anaerobes are generally resistant to this and other quinolones. Ciprofloxacin;

quinolones, inhibits DNA gyrase. Bactericidal effects are not completely reversible by

inhibitors of protein or RNA synthesis. Other quinolones and ciprofloxacin may have

multiple lethal effects. Resistance is less readily selected in vitro by ciprofloxacin than

by nalidixic acid, and single-step mutants usually remain susceptible to clinically

achievable concentrations. Resistance mediated by mutations in genes altering DNA

gyrase and expression of outer membrane proteins described for ciprofloxacin and

other quinolones. The antimicrobial spectrum and potency of ciprofloxacin, coupled

with its rapid bactericidal effects, make this fluoroquinolone a promising new

antimicrobial agent.

2.1.1.2 Enrofloxacin

A synthetic chemotherapeutic agent from the class of the fluoroquinolone carboxylic

acid derivatives. It has antibacterial activity against a broad spectrum; Gram negative

and Gram positive bacteria. Its mechanism of action is not thoroughly understood and

believed to act by inhibiting bacterial DNA gyrase, thereby prevent DNA supercoiling

and DNA synthesis. It is effective against: Pseudomonas aeruginosa, Klebsiella,

Escherichia coli, Enterobacter, Campylobacter, Shigella, Salmonella, Aeromonas,

Haemophilus, Proteus, Yersinia, Serratia, Vibrio, Brucella, Chlamydia trachomatis,

Staphylococcus, Mycoplasma and Mycobacterium. It has Variable activity against:

Streptococcus and ineffective against anaerobes.

2.1.1.3 Doxycycline

The tetracycline antibiotics group commonly used to treat a variety of infections.

Doxycycline, a semisynthetic tetracycline invented and clinically developed in the

early 1960s by Pfizer Inc.

2.1.2 Mode of action of antibiotics

Antibacterial action generally follows some of the mechanisms; inhibition or

regulation of enzymes involved in the synthesis of cell wall, nucleic acid synthesis and

repair, or protein biosynthesis. Antibiotics target the cell functioning of rapidly

dividing cells.

a. Inhibition of cell wall synthesis

Bacterial cell contains a peptidoglycan cell wall in addition to the normal inner plasma

membrane, surrounding the cellular contents. In other words, bacterial cells resemble

the primitive plant cell structure. In addition to these, gram negative bacteria also have

outer lipid bilayer. Some of the antibacterial compound interferes with the cell wall

synthesis by weakening the peptidoglycan structures in bacterial cell wall, by this

integrity of bacterial cell wall structure weakens and eventually disrupts.

b. Inhibition of nucleic acid synthesis

Antibacterial compounds interfere in the synthesis of nucleic acid of bacterial cells.

The compound quinonoles interfere with synthesis of DNA molecule by inhibiting

activity of enzyme topoisomerase. This enzyme is involved in the DNA replication.

The second generation quinolones; levofloxacin, norfloxacin and ciprofloxacin can be

used against both Gram positive and Gram negative bacteria. These compounds

specifically inhibit the bacterial topoisomease II.

c. Inhibition of protein synthesis

Some antibiotic compounds inhibit bacterial cell multiplication by inhibiting protein

synthesis. Protein synthesis is a multi step process. Majority of antibiotics inhibit the

process that occurs in 30S 0r 50S subunit of 70S bacterial ribosome. This in turn

inhibit the protein biosynthesis. Most of the antibiotics inhibits the formation of 30S

initiation complex or altogether inhibits the formation of 70S ribosome by the 30S and

50S ribosome subunits or they inhibit assembling of amino acids into a polypeptide

chain. Tetracyclines; doxycycline, block protein synthesis by preventing the binding

of aminoacyl tRNA in 30S ribosome subunit. These compounds block protein

synthesis in both prokaryotic and eukaryotic systems.

2.1.3 Harmful effect of antibiotics

Antimicrobial drug resistance is the main harmful effect of antibiotics. The principles

and definitions are stated below

1. Clinical resistance

Clinical resistance to an antimicrobial agent occurs when the MIC (Minimum

inhibitory concentration) of the drug for a particular strain of bacteria exceeds that

which is capable of being achieved with safety in vivo. Resistance to an antimicrobial

can arise:

By mutation in the gene that determines sensitivity/resistance to the agent

By acquisition of extra chromosomal DNA (plasmid) carrying a resistance

gene.

Resistance that appears after introduction of an antimicrobial agent into the

environment usually results from a selective process; the antibiotic selects for survival

of those strains possessing a resistance gene. Resistance can develop in a single step

or it can result from the accumulation of multiple mutations.

2. Cross resistance

Cross resistance implies that a single mechanism confers resistance to multiple

antimicrobial agents. Multiple resistance implies that multiple mechanisms are

involved. Cross resistance is commonly seen with closely related antimicrobial agents

and multiple resistances are seen with unrelated antimicrobial agents.

2.1.4 Human health hazard of antibiotic growth promoter

Antibiottics are often used to promote weight gain and control diseases in poultry.

More than 70% of the antibiotics used in Bangladesh are given to poultry in the

absence of infectious diseases (Mellon et al, 2001). This practice associated with

emergence of antibacterial resistant strains; Salmonella spp., Campylobacter spp.,

Escherichia coli, and Enterococcus spp. Regular and prolong administration of

antibiotics in any poultry body causes harmful effects on the physiology of that

poultry by creating antibiotic resistance as well as on human by consumption of

poultry products observed residual effects of antibiotics. Recent evidence from

scientists around the world shows that the link between the use of antibiotic growth

promoters in food animals and antimicrobial resistance is increasing (Bogaard, 1998;

Bogaard et al., 1997; Bogaard & Stobberingh, 2000; Caprioli et al., 2000; Kolár et al.,

2002). The use of growth promoting antibiotics is being placed under more pressure as

consumers increasingly fear that their use in feed rations of productive livestock leads

to the formation of resistance against bacteria pathogenic to human. Antibiotics

resistance, the world‟s most pressing public health hazards. It can cause danger and

sufferings for people who have common infections that once were easily treatable

with antibiotics. When antibiotics resistant organisms developed in a body, it requires

more expensive treatments and cause immense sufferings. Some resistant infections

can cause death. These antibiotic resistant bacteria quickly spread to family members,

school mates and co-workers threatening the community with a new strain of

infectious disease that is more difficult to cure and more expensive to treat. Up to 40%

or more of the antibiotic dose may be excreted, especially for antibiotics given at

therapeutic doses (Boxall et al., 2004). This is true for both human and animal.

However, different classes of antibiotics are more or less metabolized. Antibiotics are

excreted in urine and feces either unchanged or metabolized in the form of the

conjugated, oxidized or hydrolyzed products.

Indiscriminate use of antibiotics in all cases have widespread residual effects on edible

tissues. The use of antibiotics only in specific conditions is justified because the roll of

microbial agents is mainly to kill the rapidly dividing invading cells. There are

numerous publications showed evidence for reduction in the number of zoonotic

organisms in chicken fed antibiotic, and publications also showed increased shedding

and colonization rates of zoonotic organisms. In Europe, the DANMAP 97 report

(Bager, 1998) was the first and the most influential showing a linkage between

antibiotic growth promoter use in food animal and antibiotic resistance in pathogenic,

zoonotic, and indicator bacteria in food and human. In France, a similar approach with

cattle has been published (Martel et al., 1995). There is growing evidence that the use

of sub therapeutic antibiotic growth promoters is associated with increased anti

microbial resistance and increased risk to human health (MAFF, 1998). Sub

therapeutic usage of antibiotic growth promoters is a problem because many of the

multiple antibiotic resistant strains of bacteria are capable of passing resistance factors

to unrelated bacteria. Resistance develops when a bacterium survives exposure to an

antibiotic that normally kill the bacterial population. Usually, a mutation occurs

allowing the bacterium to survive the antibiotic exposure. Additionally, we know that

antibiotic resistance develop when there is transformation, transduction or conjugation

within a population of bacteria that allows transference of DNA leading to plasmid

formation.

2.1.5 Antibiotics as growth promoter and its effects on production

Performance and meat yield of poultry

The term „Growth Promoter‟ used for years to describe the use of sub therapeutic

levels of antibiotics to improve growth performance. It is an inappropriate term to

describe the use of antibiotic because they did not promote growth as doe‟s anabolic

hormones; growth hormone or estrogen like compounds. The general public confuses

this term with the use of anabolic hormones. The poultry industry does not use

anabolic hormones as do the swine and cattle industries. Instead of calling them

„Growth Promoters‟, they should be called „Growth Perimeter‟s because they allow

the animal to express its genetic potential for growth without compromise.

The mechanism by which antibacterial agents improve growth is not known, but

several ideas have been proposed:

(i) Nutrients are efficiently absorbed for a thinner mucous membrane small

intestinal epithelium of the gut (Boyd et al., 1967; Eyssen et al., 1963; Ford

et al., 1971; Kawai, 1980; Kawai et al., 1978; Siddons et al., 1972 and Yolton

et al., 1976)

(ii) Nutrients are spared since competing microorganisms are reduced (Eyssen, 1962,

Monson et al., 1954)

(iii) Microorganisms responsible for subclinical infections are reduced or eliminated

(Barnes et al., 1978, Eyssen, et al., 1963, Eyssen et al., 1963, Eyssen et al., 1967

and Sieburth et al., 1951)

(iv) Production of growth depressing toxins or metabolites by intestinal microflora is

reduced (Dang et al., 1960 and Huhtanen et al., 1965).

Although each proposal has merit when viewed in the light of a particular criterion,

none explains the entire growth phenomenon.

NADA (2000) reported that chlortetracycline hydrochloride is used as an aid in

stimulating growth rate and improve feed efficiency in chickens and turkeys. It has

been reported by Palic et al. (1998) that broiler diet treated with sacox and flavomycin

showed superior performance; body weight, feed convertion and mortally of poultry.

Mudric et al. (1992) used linomycin at 0 or 4mg/kg feed of broiler and concluded that

linomycin increased body weight and feed conversion efficiency. The weight gains of

chicks given linomycin and flavomycin were 4.3 and 6.4% heavier (p<0.05). While

feed conversion (FC) was better by 2.3 and 3.6% with avoparcin and lincomycin

respectively. Griffin (1979) used procaine penicillin (50mg/kg), zinc bacitracin

(25mg/kg) or nitrovin (20mg/kg) singly or in combinations. The performance of

broiler cockerels had higher in treated group than control. Giriprasad et al. (1990)

observed that the use of antibiotics in the diet resulted in less feed cost per unit body

weight of cockerels. Muzaffar et al. (2003) observed liver and abdominal fat of

broilers were not affected (p>0.05) by organic acids (0.2% genex) treatment along

with antibiotic and probiotic. Fairchild et al. (2001) reported that dietary

supplementation Bio-mos and flavomycin improved poults body weight. Mondal et al.

(1994) found the highest body weight gain for addition of flavomycin followed by

other groups. Jamroz et al. (1992) noted the 4.3 and 6.4% heaver body weight of

broiler given diet with flavomycin and lincomycin in relation to control. Izat et al.

(1990) reported diet treated with bambermycin (flavomycin, 2.2ppm) in female

broilers resulted increased dressing yield and breast and as percentage of postchill

weight in relation to those control. Garcia et al. (2002) supplemented diet apramycin

(100 ppm) and organic acidmixture (50% formic acid+50% propionic acid) at 0.0, 0.1

and 0.2% level and found that supplementation of apramycin and organic acids (0.1%)

alone increased live weight but combined did not result in a cumulative effect. It is

relevant that the extent of growth improvement elicited by sub-therapeutic levels of

growth-permittant antibiotics has not diminished substantially over years of use

(Coates et al., 1959; Hays, 1978; Heth et al., 1962). This supports the suggestion that

an infectious agent is probably not responsible for reduced growth of conventional

chicken. Other studies (Cook et al., 1954; Eyssen et al., 1962; Sieburth et al., 1954)

have failed to demonstrate changes at the genus level in the microbiological

composition of the intestinal tract when antibiotics are fed. These results strongly

implicate a role for the normal autochthonous microflora in the growth depression of

conventional animas.

2.2 Organic Acids

The acids that originate naturally called organic acids. Organic acids act as non-

therapeutic additives in poultry diets. There are several types of organic acids; acetic

acid, citric acid, lactic acid, formic acid, sorbic acid, ascorbic acid, propionic acid,

fumeric acid, malic acid etc. There are some advantages of organic acids, like acids

and its salts have long been used to preserve feed and feed stuffs and to increase the

shelf life of fermented feeds for its antimicrobial and anti fungal efficiency. In

addition to these it also lowers the pH. Organic acids also have a strong bacteriostatic

action. Organic acids now a day is being used as feed premix in poultry diets because

these acids enhance bird‟s immunity, improve response to vaccination (by raising titer

level), protein and mineral utilization, balance gut microflora, increased feed intake as

well as conversion, control diarrhea and increase productivity and decrease mortality.

Organic acids are considered to be any organic carboxylic acid including fatty acids

and amino acids, of the general structure R-COOH

Table 1. List of organic acids and their properties

Acid Chemical name Formula

Formic Formic Acid HCOOH

Acetic Acetic Acid CH3COOH

Propionic 2-Propanoic Acid CH3CH2COOH

Butyric Butanoic Acid CH3CH2CH2COOH

Lactic 2-Hydroxypropanoic Acid CH3CH(OH)COOH

Sorbic 2,4-Hexandienoic Acid CH3CH:CHCH:CHCOOH

Fumaric 2-Butenedioic Acid COOHCH:CHCOOH

HMB 2-Hydroxy-4-Methylthio Butanoic

Acid

CH3SCH3CH2CH(OH)COOH

Malic Hydroxybutanedioic Acid COOHCH2CH(OH)COOH

Tartaric 2,3-Dihydroxy-Butanedioic Acid COOHCH(OH)CH(OH)COOH

Citric 2-Hydroxy-1,2,3-

Propanetricarboxylic Acid

COOHCH2C(OH)(COOH)CH2COOH

2.2.1 Organic acids in poultry diet

High levels of production and efficient feed conversion are the need of the modern

poultry industry which to a certain extent could be achieved by the use of specific

feed additives. Antibiotic feed additives as growth promoters have long been

supplemented to poultry feed to stabilize the intestinal microbial flora, improve the

general performance, and prevent some specific intestinal pathology (Truscott et al.,

1977; Miles et al., 1984; Waldroup et al., 1995). However, because of the growing

concern over the transmission and proliferation of resistant bacteria via food chain,

the European Union (EU) in 2006 banned antibiotic growth promoters to be used as

additives in animal nutrition. The antibiotic growth promoters have been under

scrutiny for many years and have been removed from the market in many countries

(Waldroup et al., 2000). Antibiotic growth promoters and antibiotic resistance are

closely related. The increased concern on the potential for antibiotic resistant strains

of bacteria has compelled the researchers to explore the utility of other non

therapeutic alternatives; enzymes, probiotics, prebiotics, herbs, essential oils,

immune stimulants and organic acids as feed additives in poultry production. The

focus of alternative strategies has been to prevent proliferation of pathogenic bacteria

and modulation of indigenous bacteria so that the health, immune status and

performance are improved (Ravindran, 2006). The organic acids associated with

specific antimicrobial activity are short chain acids; C1-C7 and are either simple,

mono carboxylic acids; formic, acetic, propionic and butyric acids, or carboxylic

acids bearing a hydroxyl group (alpha carbon); lactic, malic, tartaric and citric acids.

Salts of some of acids have been shown to perform benefits. Other acids; sorbic and

fumaric acid are short chain carboxylic acids containing double bonds, have been

observed to possess antifungal activity (Dibner and Richards, 2005). Generally, 2

types of acidifiers are used; feed acidifiers and gut acidifiers. Feed acidifiers are pure

organic acids added to the feed by direct spraying. However, pure organic acids

corrode the GI tract of poultry, and are, also, difficult to handle. Gut acidifiers are

organic acid salts; ammonium di-propionate, potassium di-formate, sodium formate,

calcium propionate, calcium lactate, ammonium formate etc., and have little or no

corrosive effect on the gastrointestinal tract of poultry (Paul et al., 2007). Organic

acids are not antibiotics but, if used correctly along with nutritional, managerial and

bio security measures, they can be a powerful tool in maintaining the health of the

GI tract of poultry improving their zootechnical performances (Abdel-Fattah SA,

2008). If applied correctly, organic acids work in poultry, not only as a growth

promoter but also as a meaningful tool for controlling all enteric bacteria; both

pathogenic and non pathogenic (Naidu, 2000 and Wolfenden et al., 2007).

2.2.2 Mode of action of organic acids

There are different aspects of mode of action of organic acids. Beneficial effects of

organic acids are properly related to factors other than their direct antimicrobial

action. There antimicrobial activities have been reviewed by Cherrington et al., 1991)

and Russell (1992). The acids break down the DNA structure in the bacteria cell

nucleus and as a result, the bacteria cell cannot longer divide or may even die. Organic

acids are also used successfully in poultry feed to prevent digestive disorders; a result

of exaggard growth. Antimicrobial activity of organic acids examined was marginal or

absent and in the near neutral pH

region, caproic and caprylic acid reduced the

concentration of viable cells in rabbits (Skrivanova and Marounek, 2007). Ivanov

(2001) observed that organic acids reduced microbial counts. The treatment of the

contaminated litter with 5% citric acid , 4% tartaric acid and 1.5% saliayic acid

created on acid with pH under 5.0 and thus reduced the microbial counts to 2.2^ 103

.e.g. E. coli, Salmonela spp, Listeria monocytogenes, Clostidium perfringens .While

others are not; bifidobacteria and Lactobacillus spp. In poultry, the environment of the

crop with respect to microbial composition and pH

seems to be very important in

relation to the resistance to pathogens. High amount of lactobacilli and low pH

in the

crop have sown to decrease the occurrence of Salmonella in the crop ( Hinton et al.,

2000). The antibacterial activity increases with decreasing pH

value. Organic acids are

lipid soluble in the undissociated form, in which they are able to enter the microbial

cell and decrease the intracellular pH. This influence microbial metabolism, inhibiting

the action of important microbial enzymes and forces the bacterial cell to use energy

to release protons, leading to an intracellular accumulation of acid anions. The acid

anions seem to be very important regarding the antibacterial effect of organic acids

and their salts. Several investigations have shown a strong bacterial effect of organic

acids without decreasing the pH

–value in the GI tract. This is an agree with

observations that the strongest effect of organic acids with respect to digesta pH

and

antimicrobial activity are found in the stomach and the small intestine. Antibiotics

(.1% flavomycin) and organic acids (genex 0.2%) treatments decrease gram negative

bacteria counts in comparison with the basal diet (Gunal et al., 2006). Generally,

lactic acid bacteria are able to grow at relatively low pH, which means that they are

more resistant to organic acids than other bacterial species, e.g. E coli. An explanation

for this may be that gram positive bacteria have a high intracellular potassium

concentration, which provides a counteraction for the acid anions (Russell and Diez-

Gonzalez, 1998). Thus, the antibacterial effects of organic acids work through:

Modification of internal pH

Inhibition of fundamental metabolic functions

Accumulation of toxic anions

Disruption of the cellular membrane

2.2.3 Antimicrobial effect of organic acids

The undissociated organic acid pass through the cell membrane of the bacteria and

dissociate to form H+ ions which lower the pH of bacterial cell, causing the organism

to use its energy, trying to restore the normal balance. Whereas, RCOO- anions

produced from the acid can disrupt DNA, hampering protein synthesis and putting the

organism in stress. As a result the organism cannot multiply rapidly and decrease in

number (Nursey, 1997). The antibacterial effect of organic acids has been reported by

many researchers. Sofos et al. (1985) reported that the broilers on sorbic acid-

containing feed had lower coliform counts in the duodenum, lower yeast and mold

counts in the caeca, and higher bacteroides counts in the caeca. Naidu (2000),

Fushimi et al. (2001), Gunes et al. (2001), Gornowicz and Dziadek (2002),

Wolfenden et al. (2007) and Abd El-Hakim et al. (2009) concluded that organic acids

could be used in poultry as a growth promoter and a meaningful tool of controlling

intrinsic pathogenic bacteria (E. coli and Salmonella). Humphrey and Lanning (1988)

observed 0.5% formic acid resulted in reduction in the isolation rate of Salmonella

from laying hens. That also, a reduced in the incidence of infection in newly hatched

chicks. Use of organic acid mixture containing formic and propionic acid decreased

the Salmonella and lactic acid producing bacteria counts in hen‟s crop (Thompson and

Hinton, 1997). Alp et al. (1999) reported that inclusion of antibiotic and an organic

acid mixture containing lactic, fumaric, propionic, citric and formic acid separately or

in combination reduced the Enterobacteriacae count in the ileum of broilers. Ramarao

et al. (2004) observed that the total bacterial; Coliform, and Escherichia coli counts in

crop and caecal contents were lower in broilers fed gut acidifier and opined that gut

acidifier can safely replace antibacterial compounds in broiler diets with beneficial

effects on the intestinal bacterial colonization and resistance to E. coli challenge.

Moharrery and Mahzonieh (2005) described that malic acid potentiality reduced E.

coli population in the intestines of broilers. Thirumeignanam et al. (2006) reported a

decreased in total bacterial load with increase in Lactobacilli load as a result of dietary

acidification. Organic acids mixture at the level of 0.2% in the diet of broilers

decreased total bacterial and gram negative bacteria compared to the basal diet (Gunal

et al., 2006). Fumaric and sorbic acid lowered the numbers of lactic acid bacteria and

Coliforms in the ileum and caeca (Pirgozliev et al., 2007). For antimicrobial effect,

organic acids result in inhibition of intestinal bacteria leading to the reduced bacterial

competition with the host for available nutrients and diminution in the level of toxic

bacterial metabolites as a result of lessened bacterial fermentation resulting in the

improvement of protein and energy digestibility; thereby ameliorate the performance

of bird.

2.2.4 Site of action of organic acids

Organic acid exert their antimicrobial action both in feed and in the gastrointestinal

tract. Following dietary intake organic acids are only recovered from the proximal part

of the gastrointestinal tract. This is an agreement with observations that the strongest

effects of organic acids with respect to digesta pH and antimicrobial activity are found

in the stomach and the small intestine. In poultry, pathogenic bacteria; salmonella,

enter the GI tract via the crop. The environment of the crop with respect to microbial

composition and pH

seems to be very important in relation to the resistance of

pathogens. Higher amounts of Lactobacilli and lower pH

in the crop have shown to

decrease the occurrence of salmonella in the crop (Hinton et al., 2000). The

antimicrobial effect of dietary organic acids in chicken is believed to take place

mainly in the crop and gizzard. Following addition of a combination of formic and

propionic acid, high concentration of these acids could only be recovered from crop

and gizzard (Thomsom and Hinton, 1997). A study on the metabolism of dietary

added propionic acid reveals that only little dietary propionic acid reaches the lower

digestive tract and the caeca (Hume et al., 1993).

2.2.5 Organic acids on growth performance

2.2.5.1 Organic acids and live weight

Islam et al. (2008) suggested that fumaric acid (FA) may promote growth of broilers.

They showed 1.25% dietary FA had higher (p<0.05) weight gain. Higher body weight

gain was obtained for the supplementation organic acid has been reported (Denli et

al., 2003). Christian et al. (2004) observed that organic acid blend (3kg inclusion rate

per ton of feed) increased the growth of broilers under controlled conditions. The body

weight of broilers at 6 week of age was higher (p<0.05) in the groups fed diet

containing organic acid at 1kg/ton or 1.5kg/ton (Thirumeignanam et al., 2006). Paul

et al. (2007) reported that ammonium formate or calcium propionate at the level of

3g/kg feed increased the live weight at 21 day in broilers. Nezhad et al. (2007)

reported increased growth of broilers for supplementation of citric acid (0.0, 2.5% and

5.0%) with microbial phytase. Moghadam et al. (2006) administrated dietary citric

acid (0.0, 1.5% and 3.0%) and phosphorus (0.3, 0.035 and 0.4%) in broiler for a

period of 2 weeks (from 8 to 21 days) and observed increased live weight. Shen-

Guifang et al. (2005) used 0.3, 0.5 and 0.7% dietary citric acid in yellow chicken and

0.3% citric acid gave highest growth. Live weight was increased (p<0.05) by

supplementation of citric acid in Ross x Ross and Hampshire x Columbian chicks (

Rafacz-Livingston et al., 2005). The effects of supplemental organic acids and

chromium (Cr) were studied to ascertain on production and carcass traits of broilers

(Samanta et al., 2008). They concluded that, instead of individual supplementation, a

combination of Cr and organic acids may improve the production of broilers. Zhang et

al. (2005) used citric acid, fumeric and malic acid and found the mixture to support

maximum live weight in broilers. Formic acid (5,000ppm and 10,000ppm in the diet

of chicken improved (p<0.05) growth (Garcia et al., 2007). Ivanov (2005) recorded

increased live weight of broilers using lactic acid bacteria (3%), citric acid (0.7%) and

baker‟s yeast (1%). Addition of citric acid and ascorbic acids in broilers increased live

weight (Afsharmanesh and Pourreza, 2005). They also reported higher live weight by

18% with citric acid along with ascorbic acid, phytase and vitamin D3. Chitra et al.

(2004) without specifying dose reported ascorbic acid with probiotic in broilers

increased live weight. Maiorka et al. (2004) used citric acid along with fumeric, lactic

and ascorbic acids and found increased growth performance of broilers. Andrys et al.

(2003) documented highest live weight in Ross 208 male and female broilers received

the acidifier FA-30 (citric acid and phosphoric acid). Nudiens (2002) supplementing

acidifier (10 kg/ton feed) in broilers found increased live weight. Kahraman and

Bostan (1998) without specifying dose used Acid Lac Dry; citric, lactic, fumeric,

propionic and fumeric acid and/or zinc bacitracin in broilers. They reported that Acid

Dry with zinc bacitracin (p<0.05) increased live weight at 3 weeks of age. Garcia et

al. (2002) supplemented diet apramycin (100 ppm) and organic acidmixture (50%

formic acid+50% propionic acid) at 0.0, 0.1 and 0.2% level and found that

supplementation of apramycin and organic acids (0.1%) alone increased live weight

but combined did not result in a cumulative effect. Patten and Waldroup (1988)

reported that addition of 0.5 or 1.0% fumaric acid improved (p<0.01) body weights of

broilers. Skinner et al. (1991) reported that addition of 0.125% fumaric acid (p<0.05)

improved 49-day body weight of females and average weight gain of both sexes.

Kassim and Norziha (1995) reported that additive of acetic acid to diet (400 or 600

mg/kg) increased live weight. Vieira et al. (2008) reported improved body weight on

diets supplemented with a blend of organic acids (40% lactic, 7% acetic, 5%

phosphoric and 1% butyric). Owens et al. (2008) reported 12 % increase in total live

weight gain and about 9 % improvement in gain feed ratio with diets supplemented

with dietary organic acids. Improvement in live body weight by organic acid

supplementation (containing acetic acid, citric acid and lactic acid, each at 1.5 and 3.0

% in the diet) was also observed by Abdel-Fattah et al. (2008). Mazanowski et al.

(1981) noted that the addition of citric acid solution for broiler decreased live weight.

Pinchasov and Elmalich (2000) used dietary propionic and acetic acid and observed

decreased live weight with the inoculation of the acids.

2.2.5.2 Organic acids and feed intake

Moghadam et al. (2006) administrated dietary citric acid (0.0, 1.5% and 3.0%) and

phosphorus (0.3, 035 and 0.4%) in broiler for a period of two weeks (from 8 to 21

days) and observed increased feed intake. Higher feed intake for organic acid

supplementation has been observed (Denli et al., 2003). Atapattu and Nelligaswatta

(2005) noted 2% dietary citric acid increased feed intake in broilers. Zhang et al.

(2005) used citric acid along with fumeric and malic acid without specifying dose and

found the increased feed intake in broilers. Nezhad et al. (2007) used citric acid (0.0,

2.5 and 5.0%) along with microbial phytase and observed no significant effect on feed

intake of broilers.

2.2.5.3 Organic acids and feed conversion

Islam et al. (2008) showed 1.25% FA group had better (p<0.05) feed conversion (FC)

than that of groups received 5.0 and 7.5% fatty acid. Higher feed intake for organic

acid supplementation has been reported (Denli et al., 2003). The better FC noticed in

the group containing organic acid at 1 kg/ton (Thirumeignanam et al., 2006). Formic

acid (5,000ppm and 10,000ppm in the diet of chicken improved (p<0.05) FC (Garcia

et al., 2007).Vieira et al. (2008) reported improved FC with diets supplemented with

a blend of organic acids (40% lactic, 7% acetic, 5% phosphoric and 1% butyric).

Improvement in FC by organic acid supplementation (containing acetic acid, citric

acid and lactic acid, each at 1.5 and 3.0 % in the diet) was also observed by Abdel-

Fattah et al. (2008). Paul et al. (2007) reported that ammonium formate or calcium

propionate at the level of 3g/kg feed increased FC at day 21 in broilers. Afsharmanesh

and Pourreza (2005) showed citric acid and ascorbic acid, phytase, low-P (3.15g/kg),

low-Ca (7.9g/kg) and vitamin D3 increased FC in broilers. Nezhad et al. (2007) used

citric acid (0.0, 2.5 and 5.0%) and microbial phytase and observed increased FC in

broilers. Zhang et al. (2005) used citric acid and fumeric and malic acid without

specifying dose and reported the mixture to support higher FC in broilers. Arefin

(2002) without specifying dose mentioned nutrilac in chicken to improve FC. Chitra et

al. (2004) without specifying dose reported higher FC for ascorbic acid with probiotic

in broilers. Mazanowski et al. (1981) reported that the addition of citric acid solution

to the starting and finishing feeds of broiler increased FC. Kahraman and Bostan

(1998) without specifying dose used Acid Lac Dry; citric, lactic, fumeric, propionic

and fumeric acid and/or zinc bacitracin in broilers. They reported that best FC

obtained in group fed organic acid combination + zinc bacitracin. Nudiens (2002)

supplementing acidifier (10kg/ton feed) in broilers found increased FC. Shen-HuiFang

et al. (2005) using 0.3, 0.5 and 0.7% dietary citric acid in Yellow chicken observed

increased FC. Celik et al. (2003) found increased FC in broiler by supplementing

propionic acid, fumeric acid, citric acid and sorbic acid containing acidifier

(0.5kg/ton). Izat et al. (1998) without specifying dose reported buffered propionic acid

as an alternative to antibiotics improved FC of broiler. Kassim and Norziha (1995)

reported that additive of acetic acid to diet (400 or 600mg/kg) increased FC. Waldroup

(1988) reported that addition of 0.5 or 1.0% fumaric acid did not (p>0.05) influence

FC. Garcia et al. (2000) supplemented apramycin (100ppm) and organic acid mixture

(50% formic acid+50% propionic acid) at 0.0, 0.1 and 0.2% levels and found

decreased FC. Skinner et al. (1991) reported that addition of 0.125% fumaric acid had

no effect on FC. Citric acid (0.0. 0.5, 3.0%) had no significant effects on FC in

broilers (Moghadam et al.; 2006). Atapattu and Nelligaswatta (2005) reported FC was

not affected by the inclusion of 2 levels (1 and 2%) of dietary citric acid. Andrys et

al. (2003) reported that broilers ROSS 208 treated with acidifier FA-30 (citric acid

and phosphoric acid) did not affect FC.

2.2.5.4 Organic acids and survivability

Shen-HuiFang et al. (2005) using 0.3, 0.5 and 0.7% dietary citric acid in Yellow

chicken observed highest survivability in group fed 0.3% citric acid. Zhang et al.

(2005) used citric acid and fumeric and malic acid without specifying dose and found

increased survivability in broilers. Nudiens (2002) supplementing acidifier (10kg/ton

feed) in broilers found increased survivability. Chitra et al. (2004) without specifying

dose reported higher survivability for ascorbic acid with probiotic in broilers. Arefin

(2002) without specifying dose mentioned nutrilac in chicken to increase

survivability.

2.2.5.5 Organic acids and meat yield characteristics

Atapattu and Nelligaswatta (2005) used 2 levels (1 and 2%) of dietary citric acid in

broilers and observed dietary citric acid increased dressing yield. Chitra et al. (2004)

without specifying dose reported higher dressing yield fortified with ascorbic acid in

broilers. Izat et al.(1998) without specifying dose reported buffered propionic acid as

an alternative to antibiotics can be enhanced carcass quality. Nezhad et al. (2007) used

citric acid (0.0, 2.5 and 5.0%) along with microbial phytase and observed no effect on

dressing yield. Muzaffar et al. (2003) observed liver and abdominal fat of broilers

were not altered (p>0.05) by organic acids (0.2% genex), antibiotic and probiotic.

Kahraman and Bostan (1998) without specifying dose used Acid Lac Dry; citric,

lactic, fumeric, propionic and fumeric acid and/or zinc bacitracin in broilers reported

that dressing yield, liver and heart yield were not affected. The relative percentage of

abdominal fat was (p<0.05) reduced with the addition of organic acid particularly, 3%

citric acid compared to the control group (Abdel-Fattah et al., 2008). They also

reported no difference (p>0.05) among the treatments with respective to dressing

yield, liver and heart weight. Garcia et al. (2000) supplemented apramycin (100ppm)

and organic acid mixture (50% formic acid+50% propionic acid) at 0.0, 0.1 and 0.2%

levels and found decreased dressing yield at 0.1% organic acid mixture. The effects of

supplemental organic acids and chromium (Cr) were studied to ascertain on

production and carcass traits of broilers (Samanta et al., 2008). They concluded that,

instead of individual supplementation, a combination of Cr and organic acids may

improve the carcass traits of broilers more effectively presumably because of an

additive effect. Supplementation of acidifier during grower phase exhibited increased

dressing yield, leg quarters and breast meat yield in broiler (Daskiran et al., 2004). It

was concluded by waldroup et al. (1995) that feeding organic acids (lactic acid,

fumeric acid, formic acid or citric acid) to broiler is not a reliable means of controlling

carcass contamination by Salmonella. The beneficial effect of organic acids on the GI

tract was observed by Denli et al (2003) and reported organic acids to result in

remarkable increase in the intestinal weight and length of broiler chicken. Abdel-

Fattah et al. (2008) reported that the addition of any level and source of organic acids

increased feed digestion and absorption as a result of increased small intestine density

which is an indication of the intestinal villi dimension. Paul et al. (2007) reported that

the propionate, formate and lactate supplementation improved duodenal villus height.

Similar results were observed by Garcia et al. (2007) who found improved villus

height with formic acid and also greater crypt depth but villus surface area was not

influenced. The increased villus height in the small intestines has been related to a

higher absorptive intestinal surface (Loddi et al., 2004) which facilitates the nutrient

absorption and hence, has a direct impact on growth performance.

CHAPTER III

MATERIALS AND METHODS

3.1 Statement of research work

Sonali chicks were reared from day old to 8 weeks of age offered 4 antibiotics and 2

Hameco-pH

for meat purpose from 31 July to 24 September. The purpose was to

investigate the effect of organic acids as an alternative to antibiotics on growth, meat

yield and economic feasibility of rearing of sonali chicks.

3.2 Venue of the experiment

The experiment conducted at at a private poultry farm located at Godagari upazilla

under Rajshahi district.

3.3 Collection of experimental materials

Antibiotics; Ciprofloxacin, Enrofloxacin and Doxycycline with or without Organic

acids mixture (Trade name Hameco-pH

marketed by Square pharmaceuticals Limited)

were collected from local market of Bangladesh. Except doxycycline other

experimental materials were in liquid form. Doxycycline was in powder form. The

chemical composition of antibiotics and Hameco-pH are given in Table 3 and Table 2.

Table 2 Chemical composition of Hameco-pH

Chemical composition

Amount(%)

Acetic acid 14.00

Ascorbic acid 1.00

Citric acid 2.00

Lactic acid 2.00

Formic acid 15.00

Propionic acid 7.00

Sorbic acid 2.5o

Yeast extract 2.00

Ammonium format 24.00

Ammonium propionate 7.00

Propylene glycol 5.00

Water 18.50

Table 3 Chemical composition of antibiotics

Trade name Manufacturer Chemical composition

CIPRO-A VET The ACME Laboratories

Ltd.

Ciprofloxacin 10%

Enro-10 Navana Pharmaceuticals

Ltd.

Enofloxacin-10% w/v

Doxy-vet The ACME Laboratories

Ltd.

Doxycycline 100%

3.4 Experimental chicks

A total of 240 as hatched day old sonali chicks were collected from a private hatchery

(Shovon Hacthery, Baliadanga, Rajshahi). Chicks were almost uniform with regard

to body weight.

3.5 Layout of the experiment

The layout of the experiment is shown in the Table 4.There were three replications in

each dietary treatment i.e. total number of replicates was 24. Ten chicks in a pen

constituted each replicate.

Table 4 Layout showing the distribution of day-old straight run Sonali

(RIR × Fayomi) chicks to the levels and replications of the

antibiotics and Hameco-pH

.

Diet Replication Total

1 2 3 Control (without supplementation of Hemeco-p

H and

Antibiotics)

10 10 10 30

0.1% Hameco-pH + 0% Antibiotics 10 10 10 30

0% Hameco-pH +0.01% Ciprofloxacin 10 10 10 30

0% Hameco-pH +0.01% Enrofloxacin 10 10 10 30

0% Hameco-pH +0.05% Doxycycline 10 10 10 30

0.1% Hameco-pH +0.01% Ciprofloxacin 10 10 10 30

0.1% Hameco-pH +0.01% Enrofloxacin 10 10 10 30

0.1% Hameco-pH +0.05% Doxycycline 10 10 10 30

Total 80 80 80 240

3.6 Chemical composition of basal diet

All chicks were fed on a commercial feed (Kazi broiler starter) containing following

composition as declared by Kazi Farms Limited, Dhaka.

Table 5 Chemical composition of the basal diet

Nutrient Composition Amount

ME (k cal/kg) 2950

Maximum Moisture (%) 12

Crude Protein (%) 21

Fibre (%) 4.5

Crude fat (%) 6

Calcium (%) 1

Available Phosphorus (%) 0.45

Methionine (%) 0.48

Lysine (%) 1.1

Tryptophan (%) 0.19

Cystine (%) 0.4

Arginine (%) 1.20

3.7 The technique used in providing antibiotics and Hameco-p

H in the

drinking water

Antibiotics and Hameco-pH

were supplied with water. Chicks were offered 4

antibiotics (with or without Hameco-pH)

with control clean drinking water. The

purpose was to observe the interaction of antibiotics and organic acids on the growth

performance, meat yield and economic feasibility in rearing sonali chicks under farm

conditions. One group of water was kept control i.e. without supplementation of

antibiotics or organic acids. The other dietary treatments were 0.1% Hameco-pH + 0%

Antibiotics, 0% Hameco-pH + .01% Ciprofloxacin, 0% Hameco-p

H + 0.01%

Enrofloxacin, 0% Hameco-pH

+ 0.05% Doxycycline, 0.1% Hameco-pH + 0.01%

Ciprofloxacin , 0.1% Hameco-pH +.1% Enrofloxacin and 0.1% Hameco-p

H +0.05%

Doxycycline.

3.8 Management practice

The experimental chicks were exposed to similar care and management in all dietary

groups throughout the study period.

3.8.1 Preparation of house

Total of 24 pens were used for this trial having a floor space of .90m

2 per chick.

3.8.2 Litter

Rice hask was used as litter at a depth of 5cm for all the chicks.

3.8.3 Light, temperature and ventilation

During the whole experimental period all the chicks were exposed to a continuous

photo period of 23 hours and a dark period of 1 hour per day using both natural and

artificial light in an open sided house. For control of temperature and light a 60 watt

electric bulbs were used for each pen. Required temperature was tried to adjust

observing the movement of the chicks and with the help of a thermometer during first

3 weeks of age. As the experiment was conducted in summer season, most of the day

the ambient temperature was higher than the requirements (Appendix Table 2). For

this reason, last 5 weeks chicks were reared on natural temperature. Optimum

ventilation was maintained by allowing adequate air movement for preventing

dampness and minimizes harmful gasses (NH3, D2O, NO2 etc.) production.

3.8.4 Feed and water management

For the first 3 days feeds was given on news paper simultaneously a little amount of

feed was supplied in the feeder, so that the chicks were habituated to eat in the feeder.

Water was supplied in a round waterer. One feeder and one waterer were provided

each replication/pen (10 chicks).The feeders were thoroughly washed and cleaned at

the end of each week and waterers were washed twice daily.

3.8.5 Bio security

Strict bio security program was maintained during the experimental period.

3.8.6 Vaccination

The chicks were vaccinated against infectious Bursal Disease (Gumboro) and for

Newcastle Disease. Vaccines were administrated as per recommendation of

manufacturer. The vaccination schedule followed during the experimental is given in

Table 6.

Table 6 Vaccination schedule followed for the chicks

Age of

chicks

(Days)

Disease Name of

vaccine

Route of

administration and

dose

2 Newcastle Disease BCRDV One drop in each eye

12 Gumboro Izovac-3 One drop in one eye

20 Gumboro Izovac-3 One drop in one eye

21 Newcastle Disease BCRDV One drop in each eye

3.9 Record keeping and calculations

The following parameters were recorded during the eight weeks of experimental

period.

3.9.1 Live weight

Records were kept on initial live weight and live weight at 7, 14, 21, 28, 35, 42, 48

and 56 days of age for each replication. The average body weight gain of chicks in

each replication was calculated by deducting initial body weight from final body

weight.

3.9.2 Feed consumption and feed conversion

Feed consumption was recorded by deducting refusal from the supplied feed at the

end of each week per replication. Feed intake and feed conversion up to different ages

were calculated by following formula:

(a) Feed intake per week (g/chick) =Feed supplied in a week (g) - Feed weigh back in a week (g)

. Number of birds

(b) Feed conversion ratio (FCR) = Feed intake

Weight gain

3.9.3 Production cost and profitability

The production cost per chicks and per kg live weight for each treatment group was

calculated for comparison. The production cost per chick and kg live weight for each

dietary group was calculated based on the market price of the feed, cost of chicks,

antibiotics, Hameco-pH

and management cost. Management cost includes labor,

vitamins, electricity, litter and depreciation cost. Profitability per chick and kg live

weight was calculated for comparison among the dietary groups.

3.10 Slaughtering and dressing of chicks

At the end of experimental 56 days of age; one male and one female representing

average of the pen were randomly selected to have meat yield characteristics. Each

chick was slaughtered, bled, scalded, defeathered, eviscerated, dressed and dissected

and meat were stripped out of carcass to establish the meat yield (Jones, 1984).

3.11 Statistical Analysis

All growth variable included recorded and calculated were for a 4 (antibiotics) x 2

(Hameco-pH) factorial experiment with multiple observations (replications) in a

Completely Randomized Design (CRD). Analyses of variance (ANOVA) were

performed to partition variances into antibiotic (A), Hameco-pH (H), AH and error to

compare data for growth performance. All meat yield data were converted to

percentage of live weight of respective individuals prior to statistical analysis. The

meat yield data were for an A, H, sex (S), HA, AS, HS, AHS and error. ANOVA was

performed for comparison of data. Significant differences were isolated by calculating

Least Significant Difference (LSD). Genstate Computer Package was used for

statistical analysis.

Fig. 1 Showing day old sonali chicks

Fig. 2 Hameco-pH

and Antibiotics

(Enrofloxacin, Ciprofloxacin and

Doxycycline)

Fig. 3 Application of medicines in

the water

Fig. 4 Sonali chicks housed in different pens

Fig. 5 Measuring live weight of

male sonali chick

Fig. 6 Measuring live weight of

female sonali chick

Fig. 8 Recorded blood loss

Fig. 9 Recorded feather loss Fig. 10 Dissection of carcass

Fig. 7 Slaughtering chicks by Halal

method

Fig. 11 Coloured shank of

sonali chicks

A= Yellow, B= Black, C= White

D= Reddish

Fig. 12 Head

Fig. 13 Neck

Fig. 14 Drumstick meat with

bone

A B

C D

Fig. 15 Thigh meat with bone Fig. 16 Wing meat with bone

Fig. 17 Breast meat Fig. 18 Spleen

Fig. 21 Liver Fig. 22 Back

Fig. 19 Heart Fig. 20 Gizzard

CHAPTER IV

RESULTS

The results of the effects of exogenous antibiotics; 0% (control), 0.01% ciprofloxacin,

0.01% enrofloxacin, 0.05% doxycycline with or without organic acids; 0,1% Hameco-

pH

on productive performance, meat yield and cost of production are stated in this

chapter.

4.1 Growth performance

4.1.1 Live weight

The live weight of sonali chicks on different dietary antibiotics with or without

Hameco-pH

are shown in the Table 7. The data showed that the differences of live

weight against dietary antibiotic, Hameco-pH

and their combinations obtained could

not be explained by the application of those treatments (P>0.05).

4.1.2 Feed intake

No differences (p>0.05) in feed intake obtained which could be explained by dietary

antibiotics, Hameco-pH and their combinations.

4.1.3 Feed conversion

Feed conversion was highest on dietary ciprofloxacin, intermediate on control and

doxycycline and lowest on dietary enrofloxacin only at 7 days of age. At all other

ages, combinations of dietary antibiotics and its interaction with Hameco-pH

did not

make (p>0.05) any difference in feed conversion.

Table 7 Live weight (g) of Sonali chicks fed on different dietary antibiotics; 0%, 0.01% ciprofloxacin (C),

0.01% enrofloxacin (E), 0.05% doxycycline (D) with or without Hameco-pH

at different ages Age (day) Hameco-p

H

(H)

Antibiotic (A) SED and Significance+

0% C E D Mean H A HA

Initial (day

old) live

weight (g)

0% 28.70 28.23 28.47 28.40 28.45 0.225NS 0.319 NS 0.451 NS

0.1% 28.47 28.33 28.27 28.77 28.46

Mean 28.58 28.28 28.37 28.58 28.45

7 0% 63.93 63.63 61.70 61.90 62.79 0.824 NS 1.165 NS 1.647 NS

0.1% 63.40 64.50 62.87 64.43 63.80

Mean 63.67 64.07 62.28 63.17 63.30

14 0% 123.40 121.00 116.70 119.30 120.10 2.110 NS 2.990 NS 4.220 NS

0.1% 119.30 123.80 117.10 124.80 121.20

Mean 121.30 122.40 116.90 122.00 120.70

21 0% 204.00 192.50 192.00 196.60 196.30 4.540 NS 6.430 NS 9.090 NS

0.1% 196.10 203.10 187.70 207.60 198.60

Mean 200.10 197.80 189.80 202.10 197.40

28 0% 285.60 276.50 269.70 278.60 277.60 6.010 NS 8.500 NS 12.010 NS

0.1% 283.30 277.40 270.80 291.80 280.80

Mean 284.50 277.00 270.30 285.20 279.20

35 0% 386.90 368.00 361.70 378.00 373.70 8.870 NS 12.540 NS 17.730 NS

0.1% 378.90 375.60 368.20 390.50 378.30

Mean 382.90 371.80 365.00 384.30 376.00

42 0% 498.60 487.20 476.30 499.20 490.30 12.130 NS 17.150 NS 24.250 NS

0.1% 504.10 487.90 480.20 507.60 494.90

Mean 501.30 487.50 478.30 503.40 492.60

49 0% 615.20 599.00 579.50 616.50 602.60 14.970 NS 21.180 NS 29.950 NS

0.1% 624.10 607.70 591.80 622.90 611.60

Mean 619.60 603.40 585.70 619.70 607.10

56 0% 732.00 726.80 690.20 740.00 722.30 19.030 NS 26.910 NS 38.060 NS

0.1% 740.60 696.40 713.30 744.70 723.80

Mean 736.30 711.60 701.70 742.40 723.00 +NS, P>0.05; all SED`s are against 16 error degrees of freedom

Table 7 (contd.) Feed intake (g) of Sonali chicks fed on antibiotics; 0%, 0.01% ciprofloxacin (C), 0.01% enrofloxacin

(E),

0.05% doxycycline (D) with or without Hameco-pH

at different ages Age (day) Hameco-p

H

(H)

Antibiotic (A) SED and Significance+

0% C E D Mean H A HA

7 0% 52.27 48.67 51.67 50.47 50.77 1.148 NS 1.623 NS 2.296 NS

0.1% 51.15 52.10 51.97 52.10 51.83

Mean 51.71 50.38 51.82 51.28 51.30

14 0% 109.97 104.57 101.17 106.33 105.51 1.878 NS 2.657 NS 3.757 NS

0.1% 107.33 106.47 102.20 109.23 106.31

Mean 108.65 105.52 101.68 107.78 105.91

21 0% 154.50 144.70 147.70 149.60 149.10 3.04 NS 4.310 NS 6.090 NS

0.1% 148.10 152.50 144.20 152.10 149.20

Mean 151.30 148.60 145.90 150.80 149.20

28 0% 209.40 189.70 182.00 192.40 193.40 6.860 NS 9.690 NS 13.710 NS

0.1% 198.30 185.30 182.20 194.60 190.10

Mean 203.90 187.50 182.10 193.50 191.70

35 0% 219.20 213.80 227.70 228.60 222.30 7.920 NS 11.200 NS 15.830 NS

0.1% 231.70 227.70 231.10 255.00 236.40

Mean 225.50 220.80 229.40 241.80 229.40

42 0% 273.10 282.00 273.10 290.50 279.70 8.110 NS 11.460 NS 16.210 NS

0.1% 295.90 286.20 281.70 288.10 288.00

Mean 284.50 284.10 277.40 289.30 283.80

49 0% 311.20 304.30 292.30 298.70 301.60 7.940 NS 11.220 NS 15.870 NS

0.1% 319.10 311.60 305.80 324.40 315.20

Mean 315.10 308.00 299.00 311.50 308.40

56 0% 388.80 373.80 371.60 381.50 378.90 15.490 NS 21.910 NS 30.990 NS

0.1% 396.40 364.00 356.90 362.90 370.00

Mean 392.60 368.90 364.20 372.20 374.50

Cumulative

feed intake

0 - 56

0% 1719.00 1661.00 1647.00 1698.00 1681.00 35.600 NS 50.400 NS 71.300 NS

0.1% 1748.00 1686.00 1656.00 1738.00 1707.00

Mean 1733.00 1674.00 1652.00 1718.00 1694.00 +NS, P>0.05; all SED`s are against 16 error degrees of freedom

Table 7 (contd.) Feed Conversion Ratio (FCR) of Sonali chicks fed on different dietary antibiotics; 0%, 0.01% ciprofloxacin

(C), 0.01% enrofloxacin (E), 0.05% doxycycline (D) with or without Hameco-pH

at different ages Age in

week

Hameco-pH

(H)

Antibiotic (A) SED and Significance+

0% C E D Mean H A HA

7 0% 1.49 1.37 1.56 1.51 1.48 0.023 NS 0.033* 0.046 NS

0.1% 1.46 1.44 1.50 1.46 1.47

Mean 1.48 1.41 1.53 1.48 1.47

14 0% 1.85 1.83 1.84 1.86 1.84 0.037 NS 0.053 NS 0.075 NS

0.1% 1.93 1.80 1.89 1.81 1.86

Mean 1.89 1.81 1.87 1.83 1.85

21 0% 1.92 2.04 1.97 1.94 1.97 0.055 NS 0.078 NS 0.110 NS

0.1% 1.93 1.93 2.06 1.84 1.94

Mean 1.93 1.99 2.01 1.89 1.95

28 0% 2.58 2.27 2.35 2.35 2.39 0.105 NS 0.149 NS 0.211 NS

0.1% 2.78 2.53 2.20 2.32 2.33

Mean 2.43 2.40 2.27 2.33 2.36

35 0% 2.20 2.36 2.47 2.23 2.33 0.104 NS 0.147 NS 0.208 NS

0.1% 2.42 2.33 2.38 2.60 2.43

Mean 2.31 2.34 2.43 2.45 2.38

42 0% 2.46 2.38 2.38 2.42 2.41 0.087 NS 0.122 NS 0.173 NS

0.1% 2.36 2.56 2.52 2.51 2.49

Mean 2.41 2.47 2.45 2.47 2.45

49 0% 2.68 2.72 2.84 2.55 2.70 0.059 NS 0.083 NS 0.117 NS

0.1% 2.67 2.61 2.74 2.83 2.71

Mean 2.67 2.66 2.79 2.69 2.70

56 0% 3.33 2.93 3.37 3.11 3.18 0.551 NS 0.779 NS 1.101 NS

0.1% 3.41 5.32 2.95 3.04 3.68

Mean 3.37 4.13 3.16 3.08 3.43

Averag

e FCR

(0 - 56)

0% 2.45 2.38 2.49 2.39 2.43 0.040 NS 0.056 NS 0.080 NS

0.1% 2.46 2.54 2.42 2.43 2.46

Mean 2.45 2.46 2.46 2.41 2.44 +NS, P>0.05; *, P<0.05; all SED`s are against 16 error degrees of freedom

Table 7 (contd.) Production cost/kg live weight (in Taka) and mortality of sonali chicks fed antibiotics; 0%,

0.01%ciprofloxacin (C),

0.01%enrofloxacin (E), 0.05%doxycycline (D) with or without Hameco-pH

at different ages

Parameters Dietary treatments

0% H

C E D HC HE HD

*Cost (Tk )

/kg feed

38 39.60 45.90 45.60 43.40 47.50 47.20

45.00

Feed Cost (Tk)

/bird

69.96 69.26 78.89 79.05 76.24 82.84 83.67 82.07

Total Cost (Tk)

/chick

109.01 105.71 116.59 116.75 113.94 120.54 122.72 119.77

Cost (Tk)

/ kg live weight

148.92

141.77 160.40 168.83 153.97 173.08 172.12 160.82

Mortality (%) 6.66% 0% 3.33% 3.33% 3.33% 3.33% 6.66% 3.33% *, Including cost of Hameco-p

H/Antibiotic / Hameco-p

H and Antibiotic supplemented with basal diet

4.1.4 Cost of production

Production cost of sonali chicks in this study are presented in Table 7. Spending on

feed, chick, vitamins, vaccine, labour, electricity, Hameco-pH

and antibiotics were

constituted cost/chick and cost/kg live weight. The total cost of production on

different dietary antibiotics and Hameco-pH

were lowest on Hameco-pH supplemented

diet followed by control, doxycicline, ciprofloxacin, Hameco-pH with doxycicline,

enrofloxacin and Hameco-pH with enrofloxacin and highest on Hameco-p

H with

ciprofloxacin. It appeared that the total cost in different combinations of antibiotics

and Hameco-pH were mainly determined by the feed cost. Though, total cost was

almost the functions of spending on feed, it was markedly influenced by the mortality

of chicks. That was the main reason for getting lowest cost/kg live weight of chicks on

Hameco-pH

supplemented diet.

4.2 Meat yield characteristics

Effect of antibiotics, organic acids; Hameco-pH

and their combinations on meat yield

parameters of Sonali chicks during the period of 8 weeks of age is presented in

Table 8.

4.2.1 Live weight

Live weight recorded during slaughter was (Table 8) similar and higher (p<0.05)

(p>0.05) on diets control and doxycycline than those on diets ciprofloxacin and

enrofloxacin. Live weight was lower (p<0.05) and similar (p>0.05) between

enrofloxacin and ciprofloxacin diets. Males had 25.87% higher live weight than that

of their females counter parts. There was no difference (p>0.05) of Hameco-pH

and

their combinations with antibiotics on live weight of Sonali chicks.

4.2.2 Dressing yield

Chicks offered diets control and ciprofloxacin resulted similar (p<0.05) and higher

(p<0.01) dressing yield than those on received enrofloxacin and doxycycline with no

difference between lower (p>0.05) yielding enrofloxacin and doxycycline. Interaction

of antibiotics with Hameco-pH

and sex did not influence dressed yield of Sonali

chicks.

4.2.3 Total meat yield

Total meat was similar (p>0.05) and higher (p<0.01) on enrofloxacin and control

diets, where it was similar (p>0.05) and lower (p>0.05) on ciprofloxacin and

doxycycline supplemented diets. Total meat was 0.45% higher (p<0.05) in males than

that in females.

4.2.4 Breast meat

Breast meat obtained (Table 8) was similar (p>0.05) and higher (p<0.01) on

enrofloxacin and control diet than those on ciprofloxacin and doxycycline. There was

no difference between ciprofloxacin and doxycycline groups (p<0.01). Females had

0.39% higher breast meat than that of males (p<0.05). It was higher in females reared

on no supplementation of Hameco-pH. Hameco-p

H tended to reduce (p>0.05) breast

meat of Sonali chicks.

4.2.5 Dark meat

Regardless of antibiotics and Hameco-pH, males of Sonali chicks had 0.674% higher

(p<0.01) dark meat than that in females.

4.2.6 Thigh meat

Thigh meat did not differ for antibiotics, Hameco-pH, sex and their interaction

(p>0.05).

4.2.7 Drumstick meat

Difference of drumstick meat attributed to antibiotics (p<0.01) was higher in Sonali

chicks received Hameco-pH

than that on control. Above facts impressed that

antibiotics along with Hameco-pH

increased drumstick meat yield in both sexes.

4.2.8 Wing meat

Dietary antibiotics tended (p>0.05) to be depressed wing meat yield. Wing meat yield

were reduced by 0.297%, 0.31% and 0.755% for the addition of ciprofloxacin,

enrofloxacin and doxycycline to control diet respectively.

4.2.9 Edible portion

Edible portion depleted (Table 8) by 0.91%, 2.01% and 2.38% on enrofloxacin,

ciprofloxacin and doxycycline supplementation respectively than that on control.

Apparent edible portion had a tendency to be declined for adding antibiotics in control

diet. On the other hand, though insignificant (p>0.05), but Hameco-pH

had positive

effect on edible portion.

4.2.10 Abdominal fat

Abdominal fat yield obtained (Table 8) was similar (p>0.05) and higher (p<0.05) on

enrofloxacin and control diet than those on doxycycline and ciprofloxacin. However,

it was similar (p>0.05) and lower (p>0.05) on doxycycline and ciprofloxacin

supplementation. Females had 0.302% higher (p<0.01) abdominal fat than that of

males.

4.2.11 Skin weight

Skin weight was higher (p<0.05) and similar (p>0.05) on control and enrofloxacin

diets than those on doxycycline and ciprofloxacin diets. Skin weight had lower

(p<0.05) and similar (p<0.05) on doxycycline and ciprofloxacin. Skin weight was

0.78% higher in chicks offered Hameco-pH than that on control. Sex did not affect

(p>0.05) on skin weight of Sonali chicks.

4.2.12 Liver weight

Heavier (p<0.05) liver were obtained (Table 8) on enrofloxacin and doxycycline than

those on control and ciprofloxacin diets. There was no variation of liver weight which

could be explained by the antibiotic supplementation. Hameco-pH, sex and their

interactions did not alter (p>0.05) proportionate liver weight.

4.2.13 Heart weight

Antibiotics and Hameco-pH had no effect (p>0.05) on heart weight. Irrespective of

antibiotics and Hameco-pH, male chicks had a tendency to yield 0.09% higher heart

weight than that in females.

4.2.14 Gizzard weight

Heavier (p<0.05) and similar gizzard were obtained (Table 8) on enrofloxacin,

doxycycline and control diets than that on ciprofloxacin diet. Hameco-pH, sex and

their interactions did not alter proportionate liver weight (p>0.05).

4.2.15 Neck weight

There were no effects on neck weight observed for diets; antibiotics, Hameco-pH and

their combinations. Males had 0.13% higher (p<0.05) neck weight than that on

females.

4.2.16 Head weight

Diets with Hameco-pH did not alter (p>0.05) head weight of Sonali chicks. Data

(Table 8) indicated that head weight were decreased on ciprofloxacin and doxycycline

diets with no difference and higher (p<0.01) values were recorded for control and

enrofloxacin diets. Males had 0.367% heavier (p<0.01) head than that of females. Sex

did not interact with antibiotics and Hamec0-pH (p<0.05) to alter head weight.

4.2.17 Giblet

Giblet weight was higher (p<0.01) and similar (p>0.05) on enrofloxacin and control

diets than those on ciprofloxacin and doxycycline diets. Giblet weight was lower

(p<0.01) and similar (p>0.05) on ciprofloxacin and doxycycline. Chicks offered

Hameco-pH had 0.51% higher (p<0.01) giblet weight than that on control.

4.2.18 Blood loss

Irrespective of antibiotics and Hamec0-p

H female chicks had a tendency to yield

0.054% higher (p<0.05) blood loss than that on males. Antibiotics, Hameco-pH and

their combinations had no tendency (p>0.05) for alteration of blood weight.

4.2.19 Feather weight

Feather weight was increased (Table 8) by 0.56%, 0.59% and 0.62% on enrofloxacin,

doxycycline and ciprofloxacin supplementation respectively than that on control

(p<0.05). A negative apparent differences (p<0.01) were observed for adding

Hameco-pH in control diet. Irrespective of antibiotics and Hamec0-p

H female chicks

had a tendency to yield 0.78% higher (p<0.01) feather weight than that on males.

4.2.20 Drumstick bone weight

Irrespective of antibiotics and Hamec0-pH male chicks had a tendency to yield 0.234%

higher (p<0.05) feather than that on females. Diets with antibiotics, Hameco-pH

and

their combinations had no effect (p>0.05) for changing drumstick bone weight of

Sonali chicks.

4.2.21 Wing bone weight

Wing bone weight were not altered for antibiotics, Hameco-pH, sex and their

interaction (p>0.05).

4.2.22 Drumstick bone length

Males had 0.598% higher (p<0.01) drumstick bone length than that on females. Diets

on interaction of Hameco-pH

with antibiotics had a tendency to decrease drumstick

bone length than that on control.

4.2.23 Thigh bone length

Irrespective of antibiotics and Hamec0-pH male chicks had a tendency to yield 0.28%

higher thigh bone length (p<0.05) than that of females. Diets with antibiotics,

Hameco-pH

and their combinations had no effect (p>0.05) for changing thigh bone

length of Sonali chicks.

4.2.24 Wing bone length

Males had 0.507% higher (p<0.01) wing bone length than that on females. Wing bone

length depleted (Table 8) by 0.279%, 0.483% and 0.50% on enrofloxacin, doxycycline

and ciprofloxacin supplementation respectively than that on control.

Table 8 Meat yield characteristics of Sonali chicks fed on different dietary Antibiotics; 0%, 0.01% Ciprofloxacin (C),

0.01% Enrofloxacin (E) and 0.01% Doxycycline (D) with or without Hameco-pH

(H) Variable

(%)

Hameco-pH

(H)

Sex

(S)

Antibiotic (A) SED and significance+

0% C E D Mean H S A HS HA SA HSA

Live

weight

(g)

0% M 827.30 798.70 763.70 781.30 792.70 0.012NS

0.012** 0.017* 0.017NS

0.024NS

0.024NS

0.035NS

0% F 663.30 644.70 592.70 622.30 630.80

Mean 745.30 721.70 678.20 701.80 711.70

0.1% M 843.30 782.30 769.30 856.70 812.90

0.1% F 647.30 650.30 636.70 644.70 644.80

Mean 745.30 716.30 703.00 750.70 728.80

Dressing

yield

0% M 62.11 62.23 60.63 59.95 61.23 0.302NS

0.302NS

0.426** 0.426NS

0.603NS

0.603NS

0.853NS

0% F 61.71 61.24 61.95 61.03 61.48

Mean 61.91 61.91 61.29 60.49 61.35

0.1% M 62.79 61.83 60.88 60.69 61.55

0.1% F 62.71 61.42 60.82 60.75 61.43

Mean 62.75 62.75 60.85 60.72 61.49

Total

meat

yield

0% M 14.57 14.45 14.75 13.55 14.33 0.219 NS

0.219 NS

0.309** 0.309 NS

0.438 NS

0.438 NS

0.619 NS

0% F 14.14 12.92 14.47 14.00 13.88

Mean 14.35 14.35 14.61 13.77 14.11

0.1% M 14.26 13.89 14.99 13.61 14.19

0.1% F 13.74 13.51 14.35 13.33 13.73

Mean 14.00 14.00 14.67 13.7 13.96

Breast

meat

0% M 10.11 10.09 9.96 8.60 9.69 0.177 NS

0.177* 0.250** 0.250 NS

0.354 NS

0.354 NS

0.500*

0% F 10.66 8.89 10.84 10.11 10.13

Mean 10.38 10.38 10.40 9.36 9.91

0.1% M 9.88 9.13 9.79 8.94 9.44

0.1% F 9.78 9.66 10.38 9.30 9.78

Mean 9.83 9.83 10.08 9.12 9.61

Dark

meat

0% M 9.52 9.41 9.77 9.25 9.49 0.178NS

0.178** 0.252NS

0.252NS

0.356 NS

0.356 NS

0.503 NS

0% F 8.81 8.47 9.05 8.94 8.82

Mean 9.16 9.16 9.41 9.09 9.15

0.1% M 9.32 9.32 10.10 9.14 9.47

0.1% F 8.86 8.68 9.16 8.69 8.85

Mean 9.09 9.09 9.63 8.91 9.16 +NS, P>0.05; *, P<0.05; **, P<0.01; all SED`s are against 32 error degrees of freedom

Table 8 (Contd) Meat yield characteristics of Sonali chicks fed on different dietary Antibiotics; 0%, 0.01% Ciprofloxacin (C),

0.01% Enrofloxacin (E) and 0.01% Doxycycline (D) with or without Hameco-pH

(H) Variable

(%)

Hameco-pH

(H)

Sex

(S)

Antibiotic (A) SED and significance+

0% C E D Mean H S A HS HA SA HSA

Breast:

Dark meat

ratio

0% M 0.53 0.54 0.51 0.47 0.51 0.012 NS

0.012**

0.017 NS

0.017NS

0.024 NS

0.024 NS

0.034 NS

0% F 0.61 0.53 0.60 0.57 0.57

Mean 0.57 0.53 0.56 0.52 0.54

0.1% M 0.53 0.49 0.49 0.49 0.50

0.1% F 0.56 0.56 0.57 0.54 0.55

Mean 0.54 0.52 0.53 0.51 0.53

Thigh meat

0% M 7.93 7.79 8.12 7.75 7.9 0.307NS

0.307 NS

0.435 NS

0.435 NS

0.615 NS

0.615 NS

0.869 NS

0% F 7.56 7.47 8.30 7.60 7.73

Mean 7.75 7.63 8.21 7.67 7.82

0.1% M 7.72 6.12 7.80 7.62 7.32

0.1% F 7.62 6.40 7.24 7.24 7.12

Mean 6.67 6.26 7.52 7.43 7.22

Drumstick

meat

0% M 6.29 6.17 6.37 6.22 6.26 0.095 NS

0.095**

0.134**

0.134 NS

0.190** 0.190 NS

0.269 NS

0% F 5.12 5.27 5.09 5.37 5.21

Mean 5.71 5.72 5.73 5.80 5.74

0.1% M 5.37 6.65 6.24 6.93 6.30

0.1% F 4.84 5.54 5.76 5.99 5.53

Mean 5.11 6.10 6.00 6.46 5.92

Wing meat 0% M 4.02 3.66 3.84 2.99 3.63 0.111 NS

0.111* 0.157** 0.157NS

0.221NS

0.221NS

0.313 NS

0% F 3.72 2.87 3.49 3.32 3.35

Mean 3.87 3.27 3.67 3.16 3.49

0.1% M 4.04 4.19 3.90 3.26 3.85

0.1% F 4.02 3.89 3.33 3.09 3.59

Mean 4.03 4.04 3.62 3.18 3.72

Edible

portion

0% M 78.36 76.68 75.97 74.96 76.49 0.349 NS

0.349 NS

0.494** 0.494 NS

0.698 NS

0.698 NS

0.987 NS

0% F 77.80 75.30 77.51 76.37 76.75

Mean 78.08 75.99 76.74 75.67 76.62

0.1% M 78.78 76.50 78.21 76.48 77.49

0.1% F 78.11 76.54 77.69 75.75 77.02

Mean 78.45 76.52 77.95 76.11 77.26 +NS, P>0.05; *, P<0.05; **, P<0.01; all SED`s are against 32 error degrees of freedom

Table 8 (Contd) Meat yield characteristics of Sonali chicks fed on different dietary Antibiotics; 0%, 0.01% Ciprofloxacin (C),

0.01% Enrofloxacin (E) and 0.01% Doxycycline (D) with or without Hameco-pH

(H) Variable

(%)

Hameco-pH

(H)

Sex

(S)

Antibiotic (A) SED and significance+

0% C E D Mean H S A HS HA SA HSA Abdominal

fat

0% M 0.84 0.75 0.74 0.85 0.79 0.107 NS 0.107** 0.151* 0.151 NS 0.213NS 0.213 NS 0.301 NS

0% F 1.31 0.78 1.09 1.44 1.15

Mean 1.07 0.76 0.92 1.15 0.97

0.1% M 0.76 0.93 1.69 0.82 1.05

0.1% F 1.65 0.82 1.53 1.19 1.30

Mean 1.21 0.88 1.61 1.00 1.17

Skin weight 0% M 6.88 6.57 5.97 7.33 6.69 0.294* 0.294 NS 0.415* 0.415 NS 0.587 NS 0.587 NS 0.830 NS

0% F 9.46 6.50 7.26 7.28 7.62

Mean 8.17 6.53 6.62 7.30 7.16

0.1% M 8.44 7.17 8.75 7.00 7.84

0.1% F 8.68 7.81 8.14 7.55 8.04

Mean 8.56 7.49 8.44 7.28 7.94

Liver weight 0% M 1.87 1.98 2.19 1.84 1.97 0.065 NS 0.065 NS 0.092* 0.092 NS 0.130 NS 0.130 NS 0.184 NS

0% F 2.01 1.92 2.08 2.30 2.08

Mean 1.94 1.95 2.13 2.07 2.02

0.1% M 1.97 1.96 2.47 2.07 2.12

0.1% F 1.90 2.15 2.10 2.02 2.04

Mean 1.94 2.05 2.28 2.04 2.08

Neck weight

0% M 2.94 2.56 2.58 2.61 2.67 0.060 NS 0.060* 0.081 NS 0.085 NS 0.120 NS 0.120* 0.169 NS

0% F 2.26 2.38 2.70 2.73 2.52

Mean 2.60 2.47 2.64 2.67 2.60

0.1% M 2.57 2.43 2.73 2.60 2.58

0.1% F 2.37 2.40 2.69 2.43 2.47

Mean 2.47 2.42 2.71 2.52 2.53

Heart weight 0% M 0.74 0.54 0.61 0.42 0.58 0.037 NS 0.037* 0.052 NS 0.052 NS 0.074 NS 0.074 NS 0.104 NS

0% F 0.60 0.52 0.56 0.48 0.54

Mean 0.67 0.53 0.59 0.45 0.56

0.1% M 0.55 0.51 0.78 0.55 0.60

0.1% F 0.52 0.46 0.42 0.47 0.47

Mean 0.53 0.49 0.60 0.51 0.53 +NS, P>0.05; *, P<0.05; **, P<0.01; all SED`s are against 32 error degrees of freedom

Table 8 (Contd) Meat yield characteristics of Sonali chicks fed on different dietary Antibiotics; 0%, 0.01% Ciprofloxacin (C),

0.01% Enrofloxacin (E) and 0.01% Doxycycline (D) with or without Hameco-pH

(H) Variable

(%)

Hameco-pH

(H)

Sex

(S)

Antibiotic (A) SED and significance+

0% C E D Mean H S A HS HA SA HSA Head weight 0% M 4.52 3.87 4.45 4.22 4.27 0.104 NS 0.104** 0.147** 0.147 NS 0.208 NS 0.208 NS 0.294*

0% F 4.06 3.72 3.85 3.74 3.84

Mean 4.29 3.80 4.15 3.98 4.05

0.1% M 4.41 3.96 4.36 4.51 4.31

0.1% F 3.71 3.74 4.95 3.62 4.00

Mean 4.06 3.85 4.66 4.06 4.16

Giblet 0% M 16.25 14.45 15.34 15.01 15.26 0.175** 0.175 NS 0.248** 0.248 NS 0.351** 0.351 NS 0.496 NS

0% F 16.09 14.06 15.57 15.34 15.26

Mean 16.16 14.25 15.46 15.18 15.26

0.1% M 15.99 14.66 17.33 15.80 15.94

0.1% F 15.40 15.12 16.87 15.00 15.60

Mean 15.69 14.89 17.10 15.37 15.77

Blood

Weight

0% M 2.92 4.34 3.32 3.12 3.43 0.225 NS 0.225* 0.318 NS 0.318 NS 0.449 NS 0.449 NS 0.635 NS

0% F 4.00 4.43 3.07 4.11 3.91

Mean 3.46 4.39 3.20 3.62 3.67

0.1% M 3.23 3.45 3.20 3.69 3.39

0.1% F 4.14 3.91 3.84 4.12 4.00

Mean 3.69 3.68 3.52 3.90 3.70

Feather

weight

0% M 7.52 8.387 8.03 8.88 8.21 0.149** 0.149** 0.211* 0.211 NS 0.298** 0.298** 0.421*

0% F 8.53 10.20 9.57 8.07 9.09

Mean 8.02 9.29 8.80 8.48 8.65

0.1% M 7.35 6.95 7.84 7.98 7.53

0.1% F 7.86 8.21 8.05 8.68 8.20

Mean 7.61 7.58 7.94 8.33 7.87

Drumstick

bone weight

0% M 2.26 2.46 2.44 2.65 2.46 0.092 NS 0.092* 0.130 NS 0.130 NS 0.184 NS 0.184 NS 0.261 NS

0% F 2.51 2.06 2.46 2.23 2.32

Mean 2.38 2.27 2.45 2.44 2.39

0.1% M 2.61 2.38 2.86 2.41 2.57

0.1% F 2.37 2.35 2.19 2.06 2.24

Mean 2.49 2.37 2.53 2.24 2.40 +NS, P>0.05; *, P<0.05; **, P<0.01; all SED`s are against 32 error degrees of freedom

Table 8 (Contd) Meat yield characteristics of Sonali chicks fed on different dietary Antibiotics; 0%, 0.01% Ciprofloxacin

(C),

0.01% Enrofloxacin (E) and 0.01% Doxycycline (D) with or without Hameco-pH

(H) Variable

(%)

Hameco-pH

(H)

Sex

(S)

Antibiotic (A) SED and significance+

0% C E D Mean H S A HS HA SA HSA

Wing bone

weight

0% M 2.51 2.32 2.54 2.46 2.46 0.099 NS

0.099 NS

0.141 NS

0.141 NS

0.199 NS

0.199 NS

0.282 NS

0% F 2.11 2.07 2.47 2.86 2.38

Mean 2.31 2.20 2.50 2.66 2.42

0.1% M 2.68 2.64 3.12 2.41 2.71

0.1% F 2.67 2.56 2.51 2.27 2.50

Mean 2.68 2.60 2.81 2.34 2.61

Drumstick

bone length

(cm)

0% M 10.03 10.60 10.00 9.90 10.13 0.104 NS

0.104** 0.147 NS

0.147 NS

0.209* 0.209 NS

0.295*

0% F 10.17 9.27 9.68 9.27 9.60

Mean 10.10 9.93 9.84 9.58 9.87

0.1% M 10.43 9.92 9.90 10.73 10.25

0.1% F 9.67 9.65 9.33 9.70 9.59

Mean 10.05 9.78 9.62 10.22 9.92

Thigh bone

length (cm)

0% M 6.87 7.33 7.23 7.07 7.13 0.105 NS

0.105* 0.148 NS

0.148 NS

0.209 NS

0.209 NS

0.297 NS

0% F 6.93 6.53 6.82 6.87 6.79

Mean 6.90 6.93 7.03 6.97 6.96

0.1% M 7.37 6.97 7.03 7.00 7.10

0.1% F 6.93 7.02 6.97 6.57 6.87

Mean 7.15 7.00 7.00 6.78 6.98

Wing bone

length (cm)

0% M 13.47 12.50 12.83 12.80 12.90 0.136 NS

0.136** 0.193* 0.193 NS

0.272 NS

0.272 NS

0.385 NS

0% F 12.67 12.07 12.45 12.40 12.40

Mean 13.07 12.29 12.64 12.60 12.65

0.1% M 13.20 12.97 13.00 12.73 12.98

0.1% F 12.67 12.47 12.60 12.13 12.47

Mean 12.93 12.72 12.80 12.43 12.72 +NS, P>0.05; *, P<0.05; **, P<0.01; all SED`s are against 32 error degrees of freedom

CHAPTER V

DISCUSSION

5.1 Live weight

Similar live weight (p>0.05) of Sonali chicks on different dietary antibiotics with or

without Hameco-pH

(mixture of organic acids) in drinking water recorded agree with

Rahman et al. (2007); Mazanowski et al. (1981); Pinchasov and Elmalich (2000).

They concluded that the addition of organic acids at different levels did not alter the

live weight. The non significant differences on the supplementation of antibiotics,

Hameco-pH

and their combinations on live weight obtained in this study contradict

the findings of Islam et al. ( 2007); Denli et al. (2003); Thirumeignanam et al. 2006);

Paul et al. (2007); Nezhad et al. (2007); Moghadam et al. (2006); Shen-GuiFang et al.

(2005); Rafacz-Livingston et al. (2005); Zhang et al. (2005); Garcia et al. (2007);

Ivanov (2005); Afsharmanesh and Pourreza (2005); Chitra et al. (2004); Maiorka et

al. (2004); Andrys et al. (2003; Nudiens et al. (2002); Kahraman and Bostan (1998);

Patten and Waldroup (1988); Skinner et al., (1991); Kassim and Norziha (1995);

Vieira et al. (2008); Owens et al. (2008) and Palic et al. (1998); Mudric et al. (1992);

Griffin (1979); Fairchild et al. (2001) and Mondal et al. (1994). The variation resulted

might have been for the differences for genetic background, age, type, dose and

combination of organic acid and antibiotic.

5.2 Feed intake

Similar (p>0.05) feed intake on Hameco-pH noted was in support of the Nezhad et al.

(2007) and contradict Moghadam et al. (2006); Denli et al. (2003); Atapattu and

Nelligaswatta (2005) and Zhang et al. (2005).

5.3 Feed conversion

Increased feed conversion (FC) at 7 days of age resulted (p<0.05) for

supplementation of antibiotic implies that efficient absorption of nutrients through the

thinner mucous membrane of intestinal epithelium of the gut and reducing

microorganisms responsible for the competing nutrients and subclinical infection of

chick with consequently higher FC on control, ciprofloxacin and doxycycline. Non

significant improvement observed for dietary supplementation of enrofloxacin and

Hameco-pH. Positive effect (p<0.05) of antibiotic on FC catalogued is in harmony

with the result of Palic et al. (1998); Mudric et al,(1992); Kawai et al. (1980).

Insignificant (p>0.05) improvement of FC on Hameco-pH

observed (Table 7) disagree

with the findings of several authors (Islam et al., 2008; Denli et al., 2003;

Thirumeignanam et al., 2006; Vieira et al., 2008; Abdel-Fattah et al., 2008; Paul et

al., 2007; Afsharmanesh and Pourreza, 2005; Nezhad et al., 2007; Arefin, 2002;

Chitra et al., 2004; Mazanowski et al., 1981; Kahraman and Bostan, 1998; Nudiens,

2002; Shen-HuiFang et al., 2005; Celik et al., 2003 and Izat et al., 1998). All of them

stated improved FC on different levels and types of organic acids supplementation in

chicken. Insignificant (p>0.05) changes of FC of Sonali chicks on Hameco-pH

supplemented diets coincide with Skinner et al. (1991); Moghadam et al. (2006);

Atapattu and Nelligaswatta (2005) and Andrys et al. (2003).

5.4 Survivality

Relatively highest survivality obtained (Table 7) for supplementation of Hameco-pH

implies that organic acids act as antimicrobial by reducing the pathogenic organisms

responsible for various infectious diseases with consequent increased survivality. This

result contradict Shen-HuiFang et al. (2005); Zhang et al. (2005); Nudiens, (2002);

Chitra et al. (2004) and Arefin, (2002).

5.5 Meat yield characteristics

5.5.1 Dressing yield

Higher (p<0.01) dressing yield on control diet than that of antibiotic diet obtained

contradict Izat et al. (1990). They found that antibiotic; flavomycin at a rate of 2.2ppm

increased (p<0.05) dressing yield in broilers. Insignificant (p>0.05) differences of

dressing yield on dietary organic acid coincide with Kahraman and Bostan (1998) and

Abdel-Fattah et al. (2008). Dressing yield catalogued in this study contradict Atapattu

and Nelligaswatta (2005); Chitra et al. (2004); and Daskiran et al. (2004). They

showed that dietary supplementation of organic acids in broilers increased dressing

yield. The variation might be due to genetic background, age, type, dose and

combination of organic acids and antibiotics and other environments.

5.5.2 Breast meat

Similar (p>0.05) and higher (p<0.05) breast meat obtained (Table 8) on control and

ciprofloxacin supplemented diets indicated that antibiotic had no effect for improving

breast meat which contradict Izat et al. (1990). They found marked (p<0.05) effect of

dietary antibiotic (flavomycin) at a rate of 2.2ppm increased breast meat in broilers.

Hameco-pH tended to reduce (p<0.05) breast meat contradict Daskiran et al. (2004).

They exhibited increased breast meat yield in broilers on diet supplemented with

acidifier.

5.5.3 Abdominal fat

Abdominal fat measured in the present study was similar and higher (p<0.05) on

enrofloxacin and control diets viewed that antibiotic had no effect to alter abdominal

fat. The result coincides with Muzaffar et al. (2003). Non significant but increasing

trend of abdominal fat on dietary Hameco-pH contradict Abdel-Fattahet al. (2008).

They gave an idea that organic acid dietary regimens had tended to reduce (p<0.05)

abdominal fat compared with the control group in broilers.

5.5.4 Heart weight

Similar (p>0.05) heart weight found on dietary antibiotics. Hameco-pH and their

combinations in the present study. The result coincides with Abdel-Fattah et al.

(2008).

5.5.5 Liver weight

Heavier liver obtained on dietary enrofloxacin and doxycycline contradict Muzaffr et

al. (2003). They observed that liver weight was not altered by dietary antibiotic along

with organic acid. Their results also coincide with Kahraman and Bostan (1998) and

Abdel-Fattah et al. (2008).

5.5.6 Gizzard weight

Heaver (p>0.05) and similar gizzard yield found on enrofloxacin, doxycycline and

control diets than that on ciprofloxacin diet indicated that antibiotic did not alter

gizzard size. Hameco-pH could not influence (p>0.05) on gizzard size was supported

by Islam (2005). He conducted an experiment with fumeric acid on straight run

broilers and found similar (p>0.05) gizzard weight.

5.6 Economic efficiency of production

It was viewed from the obtained (Table 7) data that the total cost of production

(Tk)/kg live weight) was lowest on Hameco-pH

supplemented diet. The results

revealed the possibility for increasing economic efficiency by organic acids

supplementation on chick diet. The greatest economic efficiency on Hameco-pH

supplementation obtained about 5.05% in comparison with the control for low cost of

organic acid and highest survivality. While adding of enrofloxacin, ciprofloxacin and

doxycycline in diet increased production cost by 13.37%, 7.70% and 3.39%

respectively on comparison with the control. Greatest economic efficiency on

Hameco-pH supplementation was consistent with the findings of Soltan (2008). He

found organic acids increased economic efficiency of meat production.

CHAPTER VI

SUMMERY AND CONCLUSION

An experiment was carried out with 240 as hatched Sonali chicks at a private poultry

farm located at Godagari upazilla under Rajshahi district. The aim of the study was to

draw conclusions that organic acids (trade name Hameco-pH) might be an alternative

to antibiotics on growth performance, meat yield characteristics and economic

feasibility in rearing Sonali chicks. Chicks were fed ad libitum on a basal broiler

starter diet containing 2950 kcal ME/kg, 21% crude protein, 4.5% fibre, 6% crude fat,

1% calcium, 0.45% available phosphorus, 0.48% methionine and 1.1% lysine. The

basal diet was supplied with antibiotic (A); 0%, 0.01% ciprofloxacin (C), 0.01%

enrofloxacin (E) and 0.05% doxycycline (D) with or without Hameco-pH in drinking

water that formed 8 treatments combinations; control, 0.1% Hameco-pH (H), C, E, D,

HC, HE and HD. From the study, it was showed that productive performance; live

weight, feed intake and feed conversion did not enhance for the supplementation of

antibiotics. Moreover, production cost (cost/kg live weight) recorded on antibiotics

supplementation diets was higher than that on control for addition of extra expenditure

on antibiotics. Recorded all vital meat yield parameters; dressing yield, total meat

yield, breast meat, drumstick meat, giblet did not alter on dietary antibiotics

supplementation. On the other hand, organic acids did not mark able influence

(p>0.05) on growth performance and meat yield, but it had a tendency to increase

some growth, meat yield and profit making parameters were tended to be increased

(p>0.05). Its antimicrobial like activities tended to increase survivality resulted in

lowest production cost. The use of antibiotic was considered by some farmers as

inevitable to combat illness and productivity. But the results of this experiment

showed that there were no marked effect of antibiotics and Hameco-pH on growth,

meat yield and profitability. So, addition of antibiotics in the diet incurred additional

cost without giving any benefit. Therefore, it was concluded that Hameco-pH may be

a health friendly, safe and cost effective substitute of antibiotic. In future, various

doses of organic acid mixture and their doses are needed to be comparing to have the

most appropriate combinations and doses.

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Appendix Table 1 Meat yield (g) characteristics of Sonali chicks fed on dietary antibiotics; 0%, 0.01%

ciprofloxacin (C), 0.01% enrofloxacin (E), 0.05% doxycycline (D) with or without Hameco-pH

Sex Hameco-pH

Antibiotic Repli LVT BLW FLW HD SHWT NCWT M 0% 0% 1 798 780 713 37 38 23 M 0% 0% 2 892 861 800 39 36 27 M 0% 0% 3 792 768 710 36 35 23 F 0% 0% 1 630 610 561 24 22 15 F 0% 0% 2 667 635 573 28 24 14 F 0% 0% 3 693 665 606 29 25 16 M 0.1% 0% 1 880 860 796 44 40 22 M 0.1% 0% 2 835 809 747 37 37 22 M 0.1% 0% 3 815 780 720 31 36 21 F 0.1% 0% 1 664 645 585 23 21 16 F 0.1% 0% 2 648 622 571 27 23 15 F 0.1% 0% 3 630 595 553 22 23 15 M 0% C 1 697 661 597 26 24 19 M 0% C 2 820 790 728 33 36 20 M 0% C 3 879 842 768 34 35 22 F 0% C 1 608 585 520 22 23 15 F 0% C 2 648 615 552 25 28 15 F 0% C 3 678 648 579 25 28 16 M 0.1% C 1 818 790 734 34 34 20 M 0.1% C 2 752 726 674 30 33 18 M 0.1% C 3 777 750 695 29 35 19 F 0.1% C 1 688 665 611 27 27 18 F 0.1% C 2 619 595 544 23 24 15 F 0.1% C 3 644 615 560 23 24 14 M 0% E 1 812 785 720 36 36 21 M 0% E 2 738 710 648 37 34 20 M 0% E 3 741 720 663 29 31 18 F 0% E 1 628 609 549 24 26 17 F 0% E 2 542 520 467 24 23 15 F 0% E 3 608 595 538 20 24 16 M 0.1% E 1 777 752 691 34 35 21 M 0.1% E 2 761 740 680 32 37 22 M 0.1% E 3 770 742 682 35 31 20 F 0.1% E 1 632 608 557 31 23 17 F 0.1% E 2 688 664 607 30 25 23 F 0.1% E 3 590 565 519 33 22 12 M 0% D 1 731 707 640 30 30 20 M 0% D 2 814 790 720 35 34 20 M 0% D 3 799 774 703 34 33 21 F 0% D 1 590 560 515 20 23 16 F 0% D 2 666 645 588 27 27 18 F 0% D 3 611 586 537 23 24 17 M 0.1% D 1 857 825 757 39 39 22 M 0.1% D 2 870 830 762 43 35 25 M 0.1% D 3 843 820 751 34 42 20 F 0.1% D 1 609 584 531 22 24 15 F 0.1% D 2 675 640 580 25 30 17 F 0.1% D 3 650 630 575 23 22 15 Repli, Replication; LVT, Live weight; BLW, Blood loss weight; FLW, Feather less weight; HD, Head weight; SHWT,

Shank weight; NCWT, Neck weight;

Appendix Table 1(contd.) Meat yield (g) characteristics of Sonali chicks fed on dietary antibiotics; 0%,

0.01% ciprofloxacin (C), 0.01% enrofloxacin (E), 0.05% doxycycline (D) with or without Hameco-pH

Sex Hameco-p

H Antibiotic Repli HRT LIV GIZ SKIN ABFAT BREAST

M 0% 0% 1 8 18 21 20 4 44 M 0% 0% 2 4 13 20 37 10 41 M 0% 0% 3 6 15 19 29 7 40 F 0% 0% 1 3 13 15 34 8 34 F 0% 0% 2 4 13 12 25 10 35 F 0% 0% 3 5 14 15 35 8 37 M 0.1% 0% 1 5 19 15 41 3 43 M 0.1% 0% 2 5 16 17 35 6 42 M 0.1% 0% 3 4 15 19 31 10 40 F 0.1% 0% 1 3 15 17 23 9 34 F 0.1% 0% 2 4 12 14 28 11 32 F 0.1% 0% 3 3 10 12 33 12 29 M 0% C 1 4 16 13 21 5 34 M 0% C 2 4 15 17 30 6 43 M 0% C 3 5 16 17 28 7 44 F 0% C 1 3 13 11 17 5 27 F 0% C 2 3 11 14 24 4 29 F 0% C 3 4 13 13 22 6 30 M 0.1% C 1 4 16 15 27 9 34 M 0.1% C 2 4 15 15 28 7 34 M 0.1% C 3 4 15 16 29 6 39 F 0.1% C 1 3 16 14 25 5 30 F 0.1% C 2 3 13 13 25 6 30 F 0.1% C 3 3 13 15 26 5 34 M 0% E 1 5 17 18 20 6 40 M 0% E 2 4 15 18 27 3 38 M 0% E 3 5 18 15 21 8 36 F 0% E 1 4 13 15 23 9 34 F 0% E 2 3 11 12 16 1 32 F 0% E 3 3 13 16 26 10 30 M 0.1% E 1 6 19 17 34 13 38 M 0.1% E 2 8 19 17 30 10 37 M 0.1% E 3 4 19 16 37 16 38 F 0.1% E 1 3 13 15 26 10 33 F 0.1% E 2 3 14 19 30 16 35 F 0.1% E 3 2 13 11 22 4 31 M 0% D 1 2 14 15 26 6 30 M 0% D 2 4 15 16 31 7 37 M 0% D 3 4 14 16 29 7 34 F 0% D 1 2 13 12 21 8 33 F 0% D 2 4 16 15 25 10 30 F 0% D 3 3 14 13 22 9 31 M 0.1% D 1 5 18 20 30 7 38 M 0.1% D 2 4 15 18 33 5 42 M 0.1% D 3 5 20 22 27 9 35 F 0.1% D 1 3 12 14 23 7 28 F 0.1% D 2 2 11 17 24 10 33 F 0.1% D 3 4 16 14 26 6 29

Repli, Replication; HRT, Heart weight; LIV, Liver weight; GIZ, Gizzard weight; SKIN, Skin weight; ABFAT, Abdominal

fat; BREAST, Breast weight

Appendix Table 1(contd.) Meat yield (g) characteristics of Sonali chicks fed on dietary antibiotics; 0%,

0.01% ciprofloxacin (C), 0.01% enrofloxacin (E), 0.05% doxycycline (D) with or without Hameco-pH

Sex Hameco-

pH

Antibiotic Repli THIGH THIGHB DRUM DRUMB WING

M WINGB

M 0% 0% 1 36 7 25 9 15 11 M 0% 0% 2 31 5 28 10 19 10 M 0% 0% 3 31 6 25 9 16 10 F 0% 0% 1 27 6 16 7 12 7 F 0% 0% 2 22 4 17 9 12 7 F 0% 0% 3 26 6 18 9 13 7 M 0.1% 0% 1 38 8 24 12 16 13 M 0.1% 0% 2 32 7 22 11 17 11 M 0.1% 0% 3 28 8 22 10 18 10 F 0.1% 0% 1 22 6 16 8 12 10 F 0.1% 0% 2 32 6 16 8 14 9 F 0.1% 0% 3 20 5 15 7 13 7 M 0% C 1 29 5 21 7 12 7 M 0% C 2 30 8 26 13 16 11 M 0% C 3 34 7 27 10 16 10 F 0% C 1 26 4 15 5 6 7 F 0% C 2 21 6 18 8 12 6 F 0% C 3 25 5 18 7 10 7 M 0.1% C 1 13 8 26 11 15 11 M 0.1% C 2 29 7 25 9 16 10 M 0.1% C 3 29 7 27 8 18 10 F 0.1% C 1 17 7 18 9 14 9 F 0.1% C 2 23 6 17 7 12 8 F 0.1% C 3 22 6 19 7 12 8 M 0% E 1 33 8 26 10 16 9 M 0% E 2 26 8 23 8 14 11 M 0% E 3 34 7 24 10 14 9 F 0% E 1 26 5 16 8 11 8 F 0% E 2 21 4 16 6 10 6 F 0% E 3 27 5 13 8 10 8 M 0.1% E 1 30 9 24 11 15 12 M 0.1% E 2 30 10 26 12 17 14 M 0.1% E 3 30 8 22 10 13 10 F 0.1% E 1 23 6 18 7 10 8 F 0.1% E 2 24 8 20 8 13 9 F 0.1% E 3 22 5 17 6 9 7 M 0% D 1 27 7 22 10 12 7 M 0% D 2 33 6 26 10 12 12 M 0% D 3 31 6 25 11 11 10 F 0% D 1 22 5 17 5 10 6 F 0% D 2 26 9 17 9 11 12 F 0% D 3 23 7 16 7 10 9 M 0.1% D 1 33 6 30 10 14 10 M 0.1% D 2 35 6 28 11 16 11 M 0.1% D 3 30 6 31 10 12 10 F 0.1% D 1 22 6 18 6 9 7 F 0.1% D 2 24 6 20 9 12 8 F 0.1% D 3 24 7 20 5 9 7

Repli, Replication; THIGH, Thigh meat weight; THIGHB, Thigh bone weight; DRUM, Drumstick bone weight;

WINGM, Wing meat weight; WINGB, Wing bone weight;

Appendix Table 1(contd.) Meat yield (g) characteristics of Sonali chicks fed on dietary antibiotics; 0%,

0.01% ciprofloxacin (C), 0.01% enrofloxacin (E), 0.05% doxycycline (D) with or without Hameco-pH

Sex Hameco-pH

Antibiotic Repli TRIM BACKW ALTW ALTL THIGHB

L DRUMBL M 0% 0% 1 3 70 24 117 7 10 M 0% 0% 2 4 98 25 110 7 10.5 M 0% 0% 3 3 78 23 113 6.6 9.6 F 0% 0% 1 5 52 18 113 7 10 F 0% 0% 2 3 72 21 118 6.5 10 F 0% 0% 3 4 68 22 116 7.3 10.5 M 0.1% 0% 1 7 70 28 130 7.8 10.5 M 0.1% 0% 2 6 78 26 114 7.3 10.3 M 0.1% 0% 3 6 89 25 134 7 10.5 F 0.1% 0% 1 5 64 21 113 6.8 9.8 F 0.1% 0% 2 4 61 20 111 7 9.7 F 0.1% 0% 3 3 58 18 109 7 9.5 M 0% C 1 6 60 21 127 6.6 9.8 M 0% C 2 3 86 15 105 7.5 11 M 0% C 3 5 84 20 129 7.9 11 F 0% C 1 3 58 16 117 6.2 9.3 F 0% C 2 5 66 21 120 6.5 9 F 0% C 3 5 67 20 128 6.9 9.5 M 0.1% C 1 10 66 21 125 7 10 M 0.1% C 2 7 65 22 120 6.9 9.5 M 0.1% C 3 3 72 20 127 7 10.25 F 0.1% C 1 7 57 20 104 7 9.5 F 0.1% C 2 5 57 16 112 6.8 9.2 F 0.1% C 3 3 65 18 114 7.25 10.25 M 0% E 1 5 67 20 106 7.4 10.3 M 0% E 2 2 68 18 100 7 9.5 M 0% E 3 7 55 19 110 7.3 10.2 F 0% E 1 5 50 18 115 7.2 10 F 0% E 2 2 49 17 113 6.25 9.25 F 0% E 3 4 42 21 128 7 9.8 M 0.1% E 1 9 77 25 141 7.1 9.9 M 0.1% E 2 10 82 24 132 7 10 M 0.1% E 3 7 70 24 150 7 9.8 F 0.1% E 1 6 56 21 121 6.9 9.3 F 0.1% E 2 7 62 24 116 7 9.5 F 0.1% E 3 6 51 19 129.5 7 9.2 M 0% D 1 4 70 19 132 7.5 9.8 M 0% D 2 8 59 17 124 6.5 10 M 0% D 3 6 67 19 130 7.2 9.9 F 0% D 1 4 57 16 101 6.5 9 F 0% D 2 6 47 13 105 7.2 9.5 F 0% D 3 5 51 14 101 6.9 9.3 M 0.1% D 1 2 70 25 130 7 10.7 M 0.1% D 2 3 88 21 117 7.5 11 M 0.1% D 3 1 68 28 147 6.5 10.5 F 0.1% D 1 3 48 14 105 6.2 9.1 F 0.1% D 2 2 61 14 110 7 10 F 0.1% D 3 5 44 17 118 6.5 10

Repli, Replication; TRIM, Trim meat weight; BACKW, Back weight; ALTW, Alimentary tract weight; ALTL, Alimentary

tract length; THIGHBL, Thigh bone length; DRUMBL, Drumstick bone length;

Appendix Table 2 Recorded temperature (°C) and relative humidity (%) during

the

experimental period Date Age

(day)

6 A.M 12 P.M 6 P.M 12 A.M

Temperature

(0C)

Humidit

y

(%)

Temperature

(0C)

Humidit

y

(%)

Temperature

(0C)

Humidit

y

(%)

Temperature

(0C)

Humidit

y

(%)

31/07/11 1 24 85 25.6 88 26 86 23 84

01/08/11 2 30 87 35 79 33 87 28 83

02/08/11 3 32 80 34 78 29 81 33 75

03/08/11 4 31 79 35 81 33 77 31.5 69

04/08/11 5 29 90 30 86 31 89 29.5 91

05/08/11 6 29.8 85 31.4 83 30 86 30 82

06/08/11 7 28 87 30.5 89 32 84 26 89

07/08/11 8 30 83 30.8 92 29.9 88 27 85

08/08/11 9 32 84 31.3 88 31 90 30 81

09/08/11 10 29 87 29.7 86 29 83 30 80

10/08/11 11 28 88 28 94 27.8 95 29 88

11/08/11 12 27 97 27.6 96 25 91 28.5 93

12/08/11 13 29.6 91 29 87 28.7 92 28 89

13/08/11 14 30 86 28.9 82 30.5 79 29.8 78

14/08/11 15 31.5 84 31.5 97 31 98 32.5 82

15/08/11 16 32 92 31 94 29 89 30.5 86

16/08/11 17 25 95 30 90 28 94 26 90

17/08/11 18 26 88 33 92 27.8 93 32 76

18/08/11 19 30.2 82 27 96 30 94 28 89

19/08/11 20 32 97 33 89 34 88 29 79

20/08/11 21 31 75 32 79 29 81 28 85

21/08/11 22 32 90 34.5 83 33 86 31 88

22/08/11 23 33.3 81 31.4 81 34 82 30.5 87

23/08/11 24 32.8 82 29.3 85 31.6 81 30 81

24/08/11 25 30 79 34 87 31 89 30 74

25/08/11 26 29 89 31 91 32 93 27 94

26/08/11 27 28 88 32 89 28.5 86 28 82

27/08/11 28 29.9 91 30.5 88 27.7 92 28.5 90

Appendix Table 2 (contd.) Recorded temperature (°C) and relative humidity (%)

during the experimental period Date Age

(day)

6 A.M 12 P.M 6 P.M 12 A.M

Temperature

(0C)

Humidit

y

(%)

Temperature

(0C)

Humidit

y

(%)

Temperature

(0C)

Humidit

y

(%)

Temperature

(0C)

Humidit

y

(%)

28/08/11 29 32 82 34.5 89 34 91 30 85

29/08/11 30 29 79 32 87 30 83 31 79

30/08/11 31 31 81 35 88 33 81 32 89

31/08/11 32 29 75 36 79 32 74 27 81

01/09/11 33 33 80 33 82 31 81 29 73

02/09/11 34 28 81 35.5 81 34 77 26 87

03/09/11 35 33 82 36 84 35 85 28 80

04/09/11 36 27 76 31 80 29 82 27 76

05/09/11 37 30 85 33.8 83 30.5 80 29 81

06/09/11 38 26 82 30 82 28 83 25 78

07/09/11 39 31 80 33.5 84 31 85 27 74

08/09/11 40 29.6 87 32 91 30 93 30 86

09/09/11 41 29.5 84 29.9 89 27 81 28 80

10/09/11 42 28 90 31.8 93 29 92 28 77

11/09/11 43 34 89 35 92 35 78 32 91

12/09/11 44 28 74 35 82 33 77 31 79

13/09/11 45 34 79 32 79 32 83 29 82

14/09/11 46 31 85 33 78 30 86 28 81

15/09/11 47 30 80 34.3 85 29 80 27 78

16/09/11 48 33 95 35 87 34 88 30.5 92

17/09/11 49 27 92 29 95 31 90 25 89

18/09/11 50 31 91 32.4 96 33 94 30 93

19/09/11 51 29.5 79 34 94 31.6 87 28 94

20/09/11 52 32 90 36 93 34 91 31 90

21/09/11 53 33 92 35 91 32 89 29 90

22/09/11 54 30 87 31 89 26 85 25 82

23/03/11 55 29 88 33 88 33 75 31 85

24/09/11 56 31 84 32 84 30 88 30 86