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OCCURRENCE OF ANTIMICROBIAL RESIDUES IN BROILERS IN ENUGU METROPOLIS AND THE EFFECT OF TEMPERATURE ON THE CONCENTRATION OF OXYTETRACYCLINE RESIDUE
ABSTRACT
The high turn-over rate and quest for white meat have given more impetus to poultry production. The need to increase production and meet the demand for poultry meat has necessitated the use of veterinary drugs, especially antimicrobials, for therapeutic, prophylactic and growth promotion purposes in poultry farming. These drugs tend to accumulate in tissues and organs as residues. The presence of drug residues in tissues above maximum residue limit (MRL) becomes violative if withdrawal periods are not observed. The consumption of violative levels of antimicrobial residues could result in the development of antibiotic-resistant strains of microorganisms, allergic reaction in sensitised individuals, distortion of activities of the intestinal flora, carcinogenesis and mutagenesis. There is evidence of excessive prescription, overuse and abuse of antimicrobial drugs in veterinary practice in Nigeria, and legislation regarding drug use in veterinary practice is hardly enforced. It has become necessary to monitor the presence and level of antimicrobial residues in poultry in Nigeria using reliable screening tests. This is imperative since there is presently no established surveillance programme for detecting drug residues. Oxytetracycline (OTC) is the most widely used antimicrobial in poultry production. Although thorough cooking is part of our food culture, studies have shown that some drugs are heat stable, hence there is need to investigate the effects of heat and freezing on the stability of this drug in tissues. The study to: (i) assessed the occurrence of antimicrobial residues in broilers retailed in Enugu metropolis, (ii) evaluated the effects of cooking methods and freezing on OTC residue, (iii) compared the sensitivity of detection of OTC residues in tissues using Three Plate Test (TPT) and Premi® test. The study involved both a cross sectional survey of broilers retailed in Enugu metropolis and two experiments. The survey was an assay of antimicrobial residues in broiler meat and organs using qualitative screening methods (TPT and Premi® Test) and a quantification of tetracycline (TC) residues using specific enzyme-linked immunosorbent assay (ELISA) technique. The three major markets (Artisan, Gariki and Ogbete) in which broilers are retailed in Enugu metropolis, were used for the study. A total of 100 broilers were proportionately selected according to the sale capacity of each of the markets as follows: Artisan (40), Gariki (30) and Ogbete (30). The birds and retailers were selected using systematic random sampling technique. Muscle, liver, kidney and gizzard were collected from each of the 100 broilers for the survey. For the experiments, TPT, Premi® Test and ELISA were used to detect OTC residue in tissues of OTC treated birds and to evaluate the effects of various cooking methods (boiling, microwave grilling and roasting) and freezing time on the level of OTC residues. Fifty 5-week old broilers were raised for 3 weeks for the experiment. Four birds were sacrificed and their organs screened for residues. In the absence of residues, the remaining 46 birds were assigned into 2 equal groups (A and B). Group A birds were injected with long acting OTC at the dose of 20mg/kg body weight and group B birds were given OTC in drinking water at the dose of 4g/l for 5 days. Their organs were screened for OTC residue 24 hours after treatment. For each organ, meat juice was extracted by maceration and centrifugation. The API50 and polymerase chain reaction (PCR) were used to identify and characterize the Bacillus subtilis used for the TPT. Premi® test and ELISA were done following the manufacturer’s instructions. Graphpad Prism 5 statistical package was used to analyse the data generated. Chi-square was used to determine associations between occurrence of residues and organ types. One way analysis of variance was used to analyse other data as appropriate. Dunn’s multiple comparism was used for post hoc analysis. Three plate test inhibition zones were correlated to OTC concentration. Significance was accepted at p < 0.05. For the survey, TPT detected antimicrobial residues in 64% of broilers and the organ distribution were as follows: kidney (60%), liver (54%), gizzard (30%) and muscle (11%). Premi® test detected residues in 60% of broilers and distribution in specific organs were: kidney (49%), liver (25%) gizzard (22%) and muscle (14%). The ELISA detected tetracycline residues in 90% of broilers and detection in specific organs were: liver (96%), muscle (96%), kidney (88%) and gizzard (82%). The residue level was above MRL in 84% of liver, 100% of gizzard and 86% of muscle, whereas residues in 100% of positive kidney samples were below the MRL. For the experimental study, microwaving, boiling and roasting significantly (p < 0.05) reduced the inhibition zones produced by raw liver in TPT. There was a significant (p < 0.05) decrease in OTC residue concentration in roasted and boiled liver samples tested with ELISA. The three cooking methods had no significant (p > 0.05) effect on residue levels in muscle samples. Freezing had no significant (p > 0.05) effect on residue levels in muscle and liver samples. Three plate test had a higher sensitivity in detecting OTC residue than Premi® test in muscle and liver of birds treated by injection and those administered OTC in drinking water. There was a positive and significant correlation (r = 0.94; p < 0.05) between the TPT inhibition zones and OTC concentration.
TABLE OF CONTENTS
Title Page
Declaration i
Certification ii
Dedication iii
Acknowledgement iv
Table of Contents v
List of Tables xii
List of Figures xiii
Appendices xv
Abstract xvii
CHAPTER ONE: INTRODUCTION
Background of Study 1
Statement of the Problem 4
Research Questions 7
Study Aim and Objectives 8
Significance of the Study 9
CHAPTER TWO: LITERATURE REVIEW
2.1 Antimicrobial Agents 10
2.1.1 Definition 10
2.1.2 Antibiotics 11
2.2 Antimicrobial use in Animals 11
2.3 Classification of Antimicrobial Agents for Veterinary Use 14
2.3.1 Aminoglycosides 15
2.3.2 Beta-lactam Antimicrobial Agents 15
2.3.3 Chloramphenicol 16
2.3.4 Quinolones (Fluoroquinolones) 16
2.3.5 Lincosamides 16
2.3.6 Macrolids 16
2.3.7 Nitrofurantoin 16
2.3.8 Tetracyclines 17
2.3.9 Sulphonamides 17
2.4 Antimicrobials Banned For Use in Food Animal Production 17
2.5 Drug Residues and Related Terms 18
2.5.1 Residues 18
2.5.2 Drug residues 18
2.6 Drug Approval and Safety Evaluation of Antimicrobial Residues 19
2.7 No Effect or Maximum no Adverse Effect 20
2.8 Acceptable Daily Intake (ADI) 21
2.9 Tolerance level or Maximum Residue Limit (MRL) 22
2.10 Withdrawal Time 25
2.10.1 Withdrawal Time for Veterinary Drugs 25
2.11 Antimicrobial use in Poultry 26
2.11.1 Antimicrobials as Growth Promoters 26
2.11.2 Antimicrobials as Coccidiostats in Poultry Production 37
2.11.3 Therapeutic Antimicrobial Use in Poultry 27
2.12 Causes of Violative Level of Antimicrobial Residues in Animal Tissues 27
2.13 Public Health Hazards and Harmful Effects of Antimicrobial Residues 29
2.13.1 Drug Allergy and Hypersensitive Reaction 29
2.13.2 Effects on Human Gut Microbiota 31
2.13.3 Development of Resistance to Antimicrobial Agents 32
2.13.4 Effect on Bone Marrow/Bone Marrow Depression 33
2.13.5 Carcinogenic Effect 34
2.13.6 Industrial Effect 34
2.13.7 Other Harmful Effects 35
2.14 Prevention and Control of Antimicrobial Residue
Occurrence in Foods of Animal Origin 35
2.14.1 Disease prevention 36
Proper Diagnosis and Antimicrobial Susceptibility Testing 36
Appropriate Use of Antimicrobials and Route of Administration 37
Appropriate Dosage Regimen 37
Monitoring and Surveillance of Antimicrobial Residue 38
Antimicrobial Residues Detection and Identification 38
2.15.1 Test Matrix 39
2.16 Methods of Detection of Antimicrobial Residues in
Foods of Animal Origin 40
Microbiological Method 40
Immunochemical Methods 45
Chromatographic Methods 46
2.17. Effect of heat on Residue 47
CHAPTER THREE: GENERAL MATERIALS AND METHODS
3.1 Study Area 49
3.2 Study Design 51
3.2.1 Survey Studies 51
3.2.2 Experimental Studies 51
3.3 Sample Source, Population and Sampling Technique 51
3.3.1 Survey Studies 51
3.3.2 Experimental Study 54
3.4 Specimen Preparation 56
3.5 Data Presentation and Analysis 56
CHAPTER FOUR: ANTIMICROBIAL RESIDUES SURVEY IN COMMERCIAL BROILERS USING THREE PLATE TEST
4.1 Introduction 57
Method 59
Isolation of Bacillus subtilis 59
Identification 59
Molecular Characterization of Isolates 60
Antimicrobial Sensitivity Test 63
The Three Plate Test 64
Results 65
4.3.1 Isolation, Identification and Sensitivity of Bacillus subtilis 65
4.3.2 Sensitivity Test 70
4.3.3 Prevalence of Antimicrobial Residues in Broilers 72
4.3.4 Occurrence of Antimicrobial Residues in Broiler Meat and Organs 74
4.3.5 Organ Distribution of Antimicrobial Residues According to pH 76
4.4 Discussion 78
4.5 Conclusions 79
CHAPTER FIVE: ANTIMICROBIAL RESIDUES SURVEY INCOMMERCIALBROILERS USING PREMI® TEST
5.1 Introduction 80
5.2 Materials and method 81
5.2.1 Antibiotics Residues Detection 81
5.3 Results 82
5.3.1 Antibiotic Residues Detection in Commercial Broilers 83
5.3.2 Antibiotics Residues Detection in Sampled Organs 85
5.3.3 Detection of Antimicrobial Residues by TPT and Premi® Test 87
5.4 Discussion 89
5.5 Conclusions 90
CHAPTER SIX: DETECTION AND QUANTITATION OF TETRACYCLINE RESIDUES IN COMMERCIAL BROILERS
6.1 Introduction 91
Materials and methods 92
Organ Sample Collection And Preparation 92
Tetracycline Residues Detection 92
Results 94
Calculation of OTC Concentration from Optical Density (OD) 94
6.3.2 Organ Distribution of Tetracycline Residues 96
6.3.3 Comparison of Mean Values of the Organs with their MRL’s 98
6.4 Discussion 106
6.5 Conclusion 107
CHAPTER SEVEN: COMPARATIVE VALIDITY AND RELIABILITY OF TPT AND PREMI® TEST IN DETECTION OF OXYTETRACYCLINE RESIDUE USING ELISA AS GOLD STANDARD.
7.1 Introduction 108
7.2 Materials and Methods 110
7.2.1 Experimental Study Design 110
7.2.2 Experimental Drug Administration 110
7.2.3 Testing for OTC Residue in Experimental Birds 111
7.3 Results 111
7.3.1 TPT Detection of Oxytetracycline Residues in Group A Birds 111
7.3.2 TPT Detection of Oxytetracycline Residues in Orally Treated Birds 112
7.3.3 ELISA Detection of Oxytetracycline Residues in Muscle Samples 112
7.3.4 ELISA Detection of Oxytetracycline Residues in Liver Samples 114
7.3.5 Comparing the Detection Ability of TPT and Premi® Test in
OTC Residues Detection in Muscles of Birds in Group A 116
7.3.6 Statistical Comparison of the Validity of TPT and Premi® Test
in OTC Residue Detection In Muscles of Group A Birds 118
7.3.7 Comparing the Detection Ability of TPT and Premi® Test in
OTC Residue Detection in Muscles of Birds in Group B 121
7.3.8 Statistical Comparison of the Validity of TPT and Premi® Test in
OTC Residue Detection in Muscles of Birds in Group B 123
7.3.9 Comparing the Detection Ability of TPT and Premi® Test in
OTC Residue Detection in Liver of Birds in Group A 126
7.3.10 Statistical comparison of the validity of TPT and Premi® Test in
OTC residue detection in Liver of birds in group A 128
7.3.11 Comparing the Detection Ability of TPT and Premi® Test in
OTC Residue Detection in Liver of birds in Group B 131
7.3.12 Statistical Comparison of the Validity of TPT and Premi® Test in
OTC Residue Detection in Liver of Birds in Group B 133
7.4 Discussion 137
7.5 Conclusion 139
CHAPTER EIGHT: EFFECT OF HEATING PROCESSES (COOKING METHODS) AND FREEZING ON OTC RESIDUES IN CHICKEN MEAT AND ORGANS
8.1 Introduction 141
8.2 Materials and Methods 142
8.2.1 Sample Preparation 142
8.2.2 Effect of Heat Treatment 142
8.2.3 Effect of Freezing on OTC Residue 143
8.3 Results 143
8.3.1 Effect of Cooking Methods on OTC Residue in Muscle Using TPT 143
8.3.2 Effect of Cooking Methods on OTC Residue in Liver using the TPT 151
8.3.3 Effect of Cooking Methods on OTC Concentration in Muscle Tissues 159
8.3.4 Effect of Cooking Methods on OTC Concentration in Liver Tissues 163
8.3.5 Effect of Freezing on OTC Concentration in Both Tissues 167
8.4 Discussion 171
8.5 Conclusion 172
CHAPTER NINE: GENERAL CONCLUSIONS AND RECOMMENDATIONS
Prevalence Study with Three Plate Test (TPT) and Premi® Test 173
Organ (matrix) Distribution of Antimicrobial Residues174
Comparative Study on the Sensitivity of TPT and Premi® 174
Test in OTC Detection in Broilers
Effect of Temperature on OTC Residues in Broiler Muscle and Liver 174
REFERENCES 176
APPENDICES 205
CHAPTER ONE
INTRODUCTION
1.1 Background of Study
There is a worldwide increase in the consumption of poultry products (meat and eggs). The consumption of chicken meat has trippled over the last quarter of a century (Jordan and Partisons, 1996). Poultry products constitute a major source of animal protein in Nigeria and a source of healthy meat worldwide because of its white meat constituent. Poultry production is an important source of livelihood for rural and urban dwellers in Nigeria, as it provides employment and income. It is the most commonly kept livestock accounting for up to 70% of livestock production (Amar-Klemesu and Maxwell, 2000). The FAO report of 1988 as cited by Nwanta et al. (2012) stated that Nigeria recorded the lowest animal protein intake with an average of 6g per head per day and estimated in 2012, that in an average Nigerian meal, animal protein contributes only 3% as against 12% recommended by WHO for healthy living.
To meet up with the high animal protein demand, there is wide application of veterinary drugs in commercial poultry designed to increase the production of poultry meat and eggs. Veterinary drugs are used primarily to prevent and control infectious and non-infectious poultry diseases and assist in combating stress occasioned by vaccination, debeaking and other management practices (Dafwang et al., 1987; Kabir et al., 2003). As additives to feed and drinking water, the drugs are used for improved performance in growth especially in poultry broiler production and to promote growth and increase egg production in layer farms (Choi & Ryu, 1997; Furusawa, 1999) etc. The remnants of these drugs may remain in tissues of the animals for some time and are termed residues.
The residues of these antimicrobial drugs are excreted in body fluids of food animals and tend to accumulate in tissues/organs and eggs (Droumev, 1983: Geersema et al., 1987). Although the use of these drugs in livestock production benefit producers and consumers alike, their indiscriminate use may result in the presence of residues of these drugs in meat and other animal food products (milk and eggs) at a violative level that may be harmful to man.
The presence of drug residues in foods of animal origin is one of the most important issues in food safety because of its public health implications. They pose potential allergic reactions in sensitized individuals and alteration of the human intestinal flora which ordinarily act in competition inhibition of colonization of pathogenic bacteria (Vollard & Classener, 1994: Nisha, 2008). A link has been noted between the use of antibiotics in food animals and the development of bacterial resistance to these drugs (Stark, 2000: Reig & Toldra, 2008) and perhaps, of greater public health relevance. Other noted pathological effects produced by antimicrobial residues in food include autoimmunity, carcinogenicity, mutagenicity, bone marrow toxicity (Pavlov et al., 2008: Nisha, 2008) etc. These documented effects and others have led to the ban of some of these drugs for use in food animals by regulatory agencies. Chloramphenicol was banned because of its involvement in bone marrow toxicity while furazolidone and nitrofurans were also restricted for use in food animals because of both carcinogenic and mutagenic effects they may cause.
In the developed countries, consumer awareness of the established and potential public health implications of antimicrobial residues in food animal products and the desire of producers to avoid litigation had led to the development of several biological and chemical tests to monitor the presence, type and level of antimicrobial residues in animal tissues and products (Pennycott, 1987; Oboegbulem and Fidelis, 1996). There are several programmes, organizations and agencies concerned with food safety and drug residues in foods of animal origin. The Residue Avoidance Programme (RAP) was initiated in 1981 by the Extension Service and Food Safety and Inspection Service (FSIS) of The United States Department of Agriculture (USDA) with the goal to prevent residue through educational programme directed at the livestock industry and people serving the industry. The Ministry of Agriculture, Fisheries and Food Standards Agency (FSA) also has a mapped out strategy for the testing of chemical and antimicrobial residues in foods of animal origin. In the major food exporting countries of the world such as the USA and Canada, the European Union has well developed abattoir – based programmes for the surveillance and monitoring of antibacterial residue in meat (Kindred and Hubbert, 1993).
Antimicrobial residues in animals are conventionally detected by microbiological tests which include Bacillus stearothermophillus Disc assay(BsDA), the European Four Plate Test (FPT), the German Three Plate Test (TPT), the Premi® test, and a number of other commercial kits. These tests which are essentially qualitative are based on bacterial inhibition and are used primarily as screening tools for presence of antimicrobial residues in meat, milk and eggs. Confirmation and quantification of specific antimicrobial residues are performed by using more sensitive chromatographic and/or immunochemical methods such as validated High Performance Liquid Chromatography (HPLC) (Popelka et al, 2005) and Enzyme Linked Immunosorbent assay (ELISA).
Most available information on drug residues in foods of animal origin is mostly related to the concentration of these drugs or their metabolites in raw samples. Since most of these foods are cooked before consumption, information on the effect of heat is required to give a more accurate estimate on the concentration of these residues the consumers may be exposed to. For years, many researchers have been interested in determining whether antibiotic residues can be destroyed or concentration reduced by different cooking procedures, pasteurization, or canning processes (Rose et al., 1995; Isidori et al., 2005; Hassani et al., 2008; Hseih et al, 2011). Traditionally, heat stabilities of antibiotics have been studied based on either the evaluation of the decrease in antimicrobial activity or by specific chromatographic analysis of change in concentration after heat treatments. Relatively few studies have been carried out using both microbiological and chemical analyses in evaluating the heat stability of veterinary drug residues (Franje et al., 2010). Moreover, whether or not the heating of these compounds and any structural changes generated result in altered genotoxicity that could contribute to the mutagenicity of bacteria remains unclear.
Tetracyclines rank among the antimicrobial substances most frequently used in the animal
food production (Schmidt & Rodrick, 2003). Tetracyclines display a wide spectrum of antimicrobial action: apart from a stronger action on the gram-positive bacteria and a weaker one on the gram-negative ones, they exercise action also on mycoplasmas, chlamydiae, rickettsias, spirochetes, actinomycetes, and some protozoa (Sundin, 2003). The sum of tetracycline action is bacteriostatic.
Adverse effects on human health after the therapeutic use of tetracyclines are well known. Tetracyclines should not be used by children up to the age of 6–8 years or by pregnant women because of the risk of developing secondary tooth discoloration. Other chronic effects include nephrotoxicity, hepatotoxicity, skin hyperpigmentation in the sun exposed areas, hypersensitivity reactions. Tetracyclines have also been reported to cause hypouricemia, hypokalemia, proximal and distal renal tubular acidosis (Goldfrank et al., 2002).
1.2 Statement of the Problem
In Nigeria, as in most African countries there is excessive prescription, overuse and abuse of antimicrobial drugs in veterinary practice and human medicine. This problem is compounded by largely unrestricted availability of antimicrobial drugs and the practice of self-medication by both poultry farmers and human patients. Unauthorized and unprofessional exposure of poultry to veterinary drugs without adherence to recommended dose and/or withdrawal time ensures the accumulation of violative residues in meat and eggs.
In Enugu State, poultry products are about the cheapest source of animal protein, unlike what is obtainable in the North where there is high availability of beef and fresh milk. In the state, the fear of consuming antimicrobial contaminated milk is not much of a problem because already processed liquid or powdered milk is being used. Instead the bother lies on the poultry meat readily available to us. Poultry production is a common agricultural practice in Enugu State. There is so much reliance on veterinary drugs in the management of poultry farming, being a major source of protein and does not require much space to manage; individuals go into rearing birds to meet up with the growing human population and demand for white meat. This much reliance on drugs in poultry industry in Nigeria creates conditions that allow for development of drug resistant strains of organisms thereby encouraging the use of multiple antibiotics in the treatment of one disease condition. Potentially, this practice could give rise to accumulation of these drugs in poultry meat and eggs. In an earlier survey in the study area, violative levels of antimicrobial residues were detected in table eggs sampled from the commercial poultry farms and retail outlets. (Ezenduka et al., 2011).
In 1971, the British Government implemented The Swann committee recommendations on the use of antimicrobiasl in animal production and thus banned the use of antimicrobials meant for therapeutics for growth promotion following legistlation for use of growth promoters. Although there is legislation regarding drug use in veterinary practice in Nigeria, it is hardly if ever enforced. A good example is the ban of furazolidone and nitrofurans for use in food animals by WHO and even NAFDAC because of both carcinogenic and mutagenic effects, the drug is very much in use in this country, particularly in poultry production.
Few studies have been done in Nigeria regarding residues of veterinary drugs, some have been reported in slaughtered chicken and commercial eggs in the East (Oboegbulam and Fidelis, 1996; Ezenduka et al., 2011) and in the North by Kabir et al. (2003), Fagbamila et al. (2012) and Mbodi et al. (2014). The occurrence of residue of oxytetracycline, a commonly used antibiotic in Nigeria, was demonstrated in meat and eggs of chicken fed recommended levels of oxytetracycline in the west by Dipeolu & Alonge (2001) and Dipeolu & Dada (2005).
In the developed countries, several programmes have been mapped out with respect to food safety and drug residues in foods of animal origin. The Residue Avoidance Programme (RAP) was initiated in 1981 by the Extension Service and Food Safety and Inspection Service (FSIS) of The United States Department of Agriculture (USDA) with the goal to prevent residue through educational programme directed at the livestock industry and people serving the industry. The Ministry of Agriculture, Fisheries and Food Standards Agency (FSA) also has a mapped out strategy for the testing of chemical and antimicrobial residues in foods of animal origin. In the major food exporting countries of the world such as the USA and Canada, the European Union has well developed abattoir-based programmes for the surveillance and monitoring of antibacterial residue in meat (Kindred and Hubbert, 1993). However, in Nigeria there is no national programme in place for monitoring drug residues in food animals in farms and abattoirs.
Microbiological screening methods are preferably used for monitoring the presence of antimicrobial residues in large numbers of slaughter animals. Principally, two types of microbiological (plate and tube) tests are available. The plate tests were earlier and routinely used to detect different antimicrobials (Heitzman, 1994; Okerman et al., 2001; Jabbar, 2004). The tube tests are more exoti because of the fast growing properties of the organism at elevated temperature, it is possible to obtain an analysis result within a few hours with the test (Pikkemat et al., 2009). The use of spores instead of vegetative cells allows prolonged storage and enables commercial distribution.
Since tetracyclines are the most widely used antimicrobials in poultry production, it became necessary to gain more insight on the ability of the more convenient tube (Premi®) to detect Oxytetracycline residue in poultry tissues.
Heat treatment of food is a primary and important method for rendering edible animal products safe for human consumption. Literature abounds with the value of heat treatment in inactivating pathogenic microorganisms. Similarly, freezing is commonly done to inhibit or retard microbial growth in foods while retaining their natural constituents. There is limited information on various temperature based treatments, persistence, levels and activities of residues in edible foods and products.
There is urgent need for extensive surveillance of veterinary drug residue in poultry meat and egg in Southeastern Nigeria, where poultry production is a primary form of agriculture. In many developed countries, the public outcry against much use of antimicrobial drugs in food animals is based on the increased awareness of the public health hazards of residues of these drugs. Because there is no national programme for routine monitoring of veterinary drug residues in Nigeria and because public awareness of the adverse effects of the drug residue is low, this study became necessary to address these problems in the study area.
1.3 Research Questions
Do commercial poultry (broiler) meat and organs sold at Enugu metropolis contain antimicrobial residues, and at what proportion?
What is the prevalence of tetracycline residues and do the concentrations in different organs differ from their maximum residue limit?
Are there differences in the concentration of antimicrobial residues in chicken muscle, gizzard, liver and kidney.
What are the effects of different cooking methods (boiling, microwaving and roasting) on the concentration of oxytetracycline residue in broiler muscle and Liver?
Does freezing affect the concentration of oxytetracycline residues?
What is the validity and reliability of the two antimicrobial residue screening tests (FPT and Premi test) in OTC residue detection?
1.4 Study Aim and Objectives
Based on the questions raised above, these studies are designed to achieve the main aim of determining the occurrence of antimicrobial residues in broiler meat (muscle, gizzard, liver and kidney) in Enugu State, and the effect of freezing and cooking methods on the concentration of oxytetracycline residue. The following are the specific objectives:
To screen for the presence and determine the prevalence of antimicrobial residues in broiler meat and organs (Muscle, gizzard, liver and kidney) using the German Three Plate Test and the Premi® test kit.
To establish the relative distribution of antimicrobial residues in the muscle, gizzard, liver aznd kidney.
To determine the prevalence and relative concentration of tetracycline residues in broiler meat and organs (muscle, gizzard, liver and kidney) using Enzyme Linked Immunosorbent Assay (ELISA) immunoassay quantitative method.
To compare the validity and reliability of the microbiological methods (TPT and Premi® Test) in OTC residue detection using ELISA as gold standard.
To determine the effect of cooking methods (boiling, microwaving and roasting) on oxytetracycline residues concentration in broiler meat and organs
To determine the effect of freezing time on oxytetracycline residues concentration in broiler meat and organs
1.5 Significance of the Study.
The findings will add to baseline information on existing data on the prevalence, types and levels of veterinary drug residues in the poultry industry.
This study will provide further evidence, requested or public health education/awareness amongst stake holders as a first and necessary step towards reducing excessive application of ntimicrobials and other veterinary drugs in animal production.
The information from the study will serve as a tool for veterinarians, NAFDAC and the government in decision making and policy formation geared towards designing appropriate intervention to checkmate the occurrence of antimicrobial residues by routine screening and monitoring of food of animal origin.
The result will also educate stakeholders on the best method to process poultry meat to reduce exposure to violative level of antimicrobial residues.
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