BIOCHEMICAL CHARACTERIZATION OF ESCHERICHIA COLI ISOLATES FROM POULTRY FARMS IN UMUAHIA

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ABSTRACT

Human and animal infections caused by Escherichia coli species remain a significant public health problem worldwide. The present study was conducted to determine the occurrence of Escherichia coli in two poultry farms in Umuahia, Abia State. A total of forty-eight (48) samples (litter, droppings, water and feed) were collected from the two poultry farms and investigated  for the presence of Escherichia coli. The specimens were inoculated into peptone water which serves as pre-enrichment media and incubated aerobically at 370C for 24 hours. The peptone water culture was subsequently inoculated into Eosin methylene blue agar (EMB) and incubated aerobically at 370C for 24 hours. The isolates from EMB were characterized by cultural and biochemical tests to identify Escherichia coli. A total of twenty-four(24) Escherichia coli isolates were obtained from the forty-eight(48) samples; litter 2(4.1%) feed 6(12.50%), droppings 9(18.75%) and water 7(14.5%). The distribution of Escherichia coli isolates in the poultry locations showed that Gochiez poultry farms in Umuagu-Ibeku (Umuahia North) had nine (9) Escherichia coli isolates while Oji integrated poultry farms in Ubakala (Umuahia South) had fifteen(15) isolates. The detection of Escherichia coli isolates in poultry farms in this study poses health hazards to poultry birds and humans.





TABLE OF CONTENTS

Title page i
Certification ii
Dedication iii
Acknowledgements iv
Table of Contents v
List of Tables vi
Abstract vii

CHAPTER ONE
1.0 INTRODUCTION 1
1.1 Poultry farming Systems 2
1.2 Biochemical characterization of E.coli 5
1.3 Aims and objectives 6

CHAPTER TWO
LITERATURE REVIEW
2.1 Escherichia coli 7
2.2 Type and morphology of E.coli 8
2.3 Metabolism of E.coli 8
2.4 Culture growth of E.coli 9
2.5 Biochemical tests and identification of E.coli 10

CHAPTER THREE
MATERIALS AND METHODS
3.1 Study Area 11
3.2 Specimen collection 11
3.3 Preparation of Media 11
3.4 Isolation of Organisms 11
3.5 Identification and characterization of Escherichia coli 12
3.6 Motility test 12
3.7 Gram staining test 12
3.8 Biochemical tests 13
3.8.1 Indole test 13
3.8.2 Citrate utilization test 13
3.8.3 Hydrogen sulphide test 14

CHAPTER FOUR
RESULTS 15

CHAPTER FIVE
5.1 DISCUSSION, CONCLUSION AND RECOMMENDATION
References





LIST OF TABLES

Table     Title           Page
  1     Colonial morphology and biochemical characteristics of isolates 16

  2   Isolation of Escherichia coli from different specimen types in the poultry farms studied  17

  3    Distribution of Escherichia coli isolates in the two poultry farms 18



                                                        


CHAPTER ONE

1.0 INTRODUCTION
The poultry is a significant economic force in the United States. The national agricultural statistics service estimates that the average value of poultry production is more than billion dollars per year. The demand for chicken has increased over the past 50 years from one million pounds in 2000, a 4-log10 increase in production. In Nigeria, poultry food is used as an alternative protein source due to their relative cheaper prices as compared to other sources such as cattle, goat, pig and sheep. (Sago et al., 2012).

Poultry provides globally important sources of animal protein and are amongst the most intensively reared of all livestock species. Diseases of poultry are therefore of major concern, both locally and on an international scale. Poultry production have been brought to the edge, because of the large numbers of infectious diseases outbreaks that strikes the poultry farm from time to time in the absence of good hygiene. It is also important to consider the concept of, and the need for, biosecurity or range of management procedures designed to protect livestock from infection. (Webster, 2010).

Escherichia coli is one of the common microbial Flora's of gastrointestinal tract of poultry and human beings including other animals but may become pathogenic to both. (Amigo et al., 2011).

According to Talaro et al. (2012) among the diseases caused by Escherichia coli, some are often severe and sometimes lethal infections such as menigitis, urinary tract infections, epidemic diarrhea of adults and children and yolk sac infections. Escherichia coli is the primary causative agent of cellulitis, septicemia and air sacculitis in poultry and the most significant bacterial pathogen of broiler chicken. During the past 2 decades, severe outbreaks of gastrointestinal illness have occurred by food borne pathogenic Escherichia coli especially 0157:H7.

1.1 POULTRY FARMING SYSTEM
Poultry farming is the process of raising domesticated birds such as chickens, ducks, turkeys and geese for the purpose of farming meat or eggs for food. Poultry are farmed in great numbers with chickens being the most numerous. More than 50 billion chickens are raised annually as a source of food, for both their meat and their eggs. Chickens raised for eggs are usually called layers while chickens raised for meat are often called broilers.(Defra 2011) In the US, the national organization overseeing poultry production is the Food and Drug Administration (FDA). In the UK, the national organisation is the Department for Environment, Food and Rural Affairs (Defra).

According to the researchers and scientists, 74 percent of the world's poultry meat, and 68 percent of eggs are produced in ways that are described as 'intensive'.(Prafulla, R 2015) One alternative to intensive poultry farming is free-range farming using lower stocking densities. Poultry producers routinely use nationally approved medications, such as antibiotics, in feed or drinking water, to treat disease or to prevent disease outbreaks. Some FDA-approved medications are also approved for improved feed utilization.(Appleby M.C et al., 2010)

Commercial hens usually begin laying eggs at 16–20 weeks of age, although production gradually declines soon after from approximately 25 weeks of age. This means that in many countries, by approximately 72 weeks of age, flocks are considered economically unviable and are slaughtered after approximately 12 months of egg production,(Tactacan, G.B et al.,  2010) although chickens will naturally live for 6 or more years. In some countries, hens are force moulted to re-invigorate egg-laying.

Environmental conditions are often automatically controlled in egg-laying systems. For example, the duration of the light phase is initially increased to prompt the beginning of egg-laying at 16–20 weeks of age and then mimics summer day length which stimulates the hens to continue laying eggs all year round; normally, egg production occurs only in the warmer months. Some commercial breeds of hen can produce over 300 eggs a year. (Lambert T. 2018).

Free-range poultry farming allows chickens to roam freely for a period of the day, although they are usually confined in sheds at night to protect them from predators or kept indoors if the weather is particularly bad. In the UK, the Department for Environment, Food and Rural Affairs (Defra) states that a free-range chicken must have day-time access to open-air runs during at least half of its life. Unlike in the United States, this definition also applies to free-range egg-laying hens. The European Union regulates marketing standards for egg farming which specifies a minimum condition for free-range eggs that "hens have continuous daytime access to open-air runs, except in the case of temporary restrictions imposed by veterinary authorities".(Allison L. 2018) The RSPCA "Welfare standards for laying hens and pullets" indicates that the stocking rate must not exceed 1,000 birds per hectare (10 m2 per hen) of range available and a minimum area of overhead shade/shelter of 8 m2 per 1,000 hens must be provided.

Free-range farming of egg-laying hens is increasing its share of the market. Defra figures indicate that 45% of eggs produced in the UK throughout 2010 were free-range, 5% were produced in barn systems and 50% from cages. This compares with 41% being free-range in 2009 (Ranger 2011).

Suitable land requires adequate drainage to minimize worms and coccidial oocysts, suitable protection from prevailing winds, good ventilation, access and protection from predators. Excess heat, cold or damp can have a harmful effect on the animals and their productivity. (Deeb N et al., 2010). Free-range farmers have less control than farmers using cages in what food their chickens eat, which can lead to unreliable productivity, though supplementary feeding reduces this uncertainty. In some farms, the manure from free-range poultry can be used to benefit crops.

The benefits of free-range poultry farming for laying hens include opportunities for natural behaviours such as pecking, scratching, foraging and exercise outdoors. (Deeb N et al., 2010)

Both intensive and free-range farming have animal welfare concerns. Cannibalism, feather pecking and vent pecking can be common, prompting some farmers to use beak trimming as a preventative measure, although reducing stocking rates would eliminate these problems. Diseases can be common and the animals are vulnerable to predators. Barn systems have been found to have the worst bird welfare.(Sherwin C et al., 2010) In South-East Asia, a lack of disease control in free range farming has been associated with outbreaks of Avian influenza.(WSPA 2013)

Free-Run
Instead of keeping them in cages, free-run laying hens roam freely within an enclosed barn. This type of housing also provides enrichment for the hens, including nesting boxes and perches that are often located along the floor of the barn. Many believe that this type of housing is better for the bird than any caging system, but it has its disadvantages, too. Due to the increase in activity of the birds, dust levels tend to elevate and the air quality decreases. When air quality drops, so does production as this compromises the health and welfare of both birds and their caretakers. (David B et al., 2015)

1.2 BIOCHEMICAL CHARACTERIZATION OF Escherichia coli
Escherichia coli described by Theoder Escherichia in 1885, member of family Enterobacteraceae, a short Gram negative, non-spore forming and usually peritrichous and fimbriate bacillus. A capsule or microcapsule is often present and a few strains produced profuse polysaccharide slime. Its present name Escherichia coli were officially accepted in 1958 in honor of its discoverer (Kuhnert et al., 2010). E.coli is a facultative anaerobe that can be recovered from samples on general or selective media at 37°C under aerobic conditions; growth of these organisms has also been observed at 44°C. E.coli is catalase positive, oxidase negative, non-spore forming and rod shaped bacterium. When growing anaerobically there is an absolute requirement for fermentable carbohydrate. Glucose is fermented to give acid and gas. These organisms are able to utilize carbon and nitrogen sources for all their metabolic and energy needs. Almost all strains of E.coli are reported to produce acid from glucose, lactose, mannitol and arabinose but not from adonitol and inositol and acid from dulcitol, salicin and sucrose varies from different strains. Most strains of E.coli are non-pathogenic (Stender et al., 2010).

However, some strains differ from commensals in that they express virulence factors directly involved in pathogenesis there by causing disease. (Schroeder et al., 2012). E.coli cause disease in chickens collectively known as avian pathogenic E.coli i.e APEC (Kariuki et al., 2011).

1.3 AIMS AND OBJECTIVES
Isolation of bacteria from poultry and its environments.

To characterize and evaluate the different organisms isolated

To determine the biochemical characterization of the isolates.
 

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