ABSTRACT
This study was done to determine isolates from five brand of fish feeds (Top, Eco, Alleraqua, Blue crown and Nsun). The samples were collected from Ndoro market, and Umuahia main market, with sterile containers. A total of 50 samples were collected from the two different markets in Umuahia, and was analysed in the laboratory by standard microbiological methods. A total of 38 bacteria (Staphylococcus aureus, E.coli, Bacillus substilis, Salmonella spp, Proteus mirabilis, Shigella spp and Klebsilla spp) were isolated from the fish feed samples. Results showed that Eco fish feed had the highest bacterial load of 7.2 x 105cfu/g, E.coli had the highest prevalence of 35% in fish feed, Results showed that the fish feed sold at Ndoro, Umuahia in Abia State has the highest occurrence of bacteria in them. The enzymatic activities of the feed in the results proved that the protease and lipase activities are essential for the utilization of protein and lipid in Shigella spp and E.coli ranging from 0.962 ± to 1.224 ± and 2.058 ± to 2.307 ± respectively. These corresponds to the high growth of the fishes. Amylase activity proved the need for the hydrolysis of carbohydrates respond to the level of dietary carbohydrate to starch - rich feed. Also in the study of the antibiotic test showing the presence of resistance strains in fish feed, it was observed that S.typhi isolates were resistant to tetracycline and erythromycin. The antibiotics (ciprofloxacin, norfloxacine, gentamycin, amoxil, streptomycin, erythromycin, ampiclox etc.) used in this study are commonly used clinically in Umuahia to treat infected humans. Feeds contaminated with pathogenic microbes such as Salmonella, should be decontaminated before use or destroyed out rightly.
TABLE OF CONTENTS
Title i
Dedication ii
Acknowledgement iii
Table of contents iv
List of tables vi
List of figures vii
Abstract viii
CHAPTER ONE
INTRODUCTION
1.1 Background of the Study 1
1.2 Statement of the Problem 2
1.3 Objectives of the Study 2
CHAPTER TWO
LITERATURE REVIEW
2.1 Definition of Fish Feed 3
2.1.1 Types of Fish Feed 3
2.2 Nutrition Value Fish Feed 6
2.2.1 Proteins and Amino Acids 6
2.2.2 Lipids 7
2.2.3 Carbohydrates 7
2.2.4 Vitamins and Minerals 7
2.2.5 Pigment 7
2.2.6 Binding Agents 8
2.2.7 Preservatives 8
2.2.8 Attractants 8
2.2.9 Other Feed stuffs 8
2.3 Feed Preparation 9
2.4 Bacteria associated with Fish Feed 10
2.5 Bacteria associated with farmed fish 11
2.6 Factors Contributing to Fish Contamination 13
2.7 Antimicrobial Resistance 14
CHAPTER THREE
MATERIALS AND METHODS
3.1 Sample Collection 16
3.2 Media Preparation 16
3.2.1 Enumeration and Isolation of Bacteria from Fish Feed Sample 16
3.2.2 Identification of Isolates 16
3.3 Gram Staining 17
3.4 Motility Test by Hanging Drop Method 17
3.5 Biochemical Tests 18
3.5.1 Catalase Test 18
3.5.2 Coagulase Test (Slide test) 18
3.5.3 Oxidase Test 18
3.5.4 Citrate Utilization Test 19
3.5.5 Indole Test 19
3.5.6 Voges-Proskauer Test 20
3.5.7 Methyl Red Test 20
3.6 Purification of Isolate 21
3.7 Qualitative Screening of Enzyme Producing Isolates 21
3.7.1 Purification of Enzyme Isolates 21
3.7.2 Production of Amylase Enzyme 21
3.7.3 Production of Lipase Enzyme 21
3.7.4 Screening of Isolates for the Pectinase Activity 22
3.8 Antimicrobial Susceptibility Tests 22
CHAPTER FOUR
RESULTS AND DISCUSSION
4.1 Results 24
4.2 Discussion 33
CHAPTER FIVE
CONCLUSION
5.1 Conclusion 37
REFERENCE
LIST OF TABLES
Table Title Page
4.1 Identification and Characterize of Bacterial Isolate from Fish Feed Samples 25
4.2 Mean Microbial Count from Fish Feed Sample 26
4.3 Distribution of Bacterial Isolates from Fish Feed Samples 29
4.4 Percentage occurrence of Bacterial isolates from fish feed samples 30
4.5 Enzymatic activities and zone of inhibitions 31
4.6 Antibiotic susceptibility tests of bacterial isolates from fish feed samples 32
CHAPTER ONE
1.1 INTRODUCTION
Fish feed may serve as a substrate for a wide variety of microorganisms (Mansong and Ubong, 2016). Some of the micro flora are adapted to the desiccated conditions in soil and are transferred by insects, dust and wind to similar niche where they are capable of degrading organic matter and/or exist in a survival state until the moisture is high enough for bacterial action. While other microorganisms, primary moulds, actively grow within stored seeds including the low amount of available moisture as substrates (Crump et al., 2002). The chemical and nutritional constituents of fish feeds are important for fish nutrition and growth, but are only part of the animal feed matrix. Feed may contain diverse micro flora that is acquired from multiple environmental sources, including dust, soil, water and insects. Feed materials may be inoculated at any time during growing, harvesting, processing, storage and dispersal of the feed. According to Maciorowskiet al., (2004), micro flora found in feed materials may come from a variety of ecological niches, which may include soil and gastrointestinal tracts, and have to adapt to the conditions found in fish feed and feed components in order to survive. The microbial diversity found in different feeds is dependent on the water activity, oxygen tension, pH and nutrient composition of the feed matrix. Micro flora can decrease grain value through nutritional changes, physical damage or the production of toxins deleterious to the health.
Different types of feeds are given to fish depending on the purpose, examples are Durante, Coppens, Aqua,Multi etc. The percentage of each ingredient in the feed varies depending on the type of feed. The ingredient include ground yellow corn, ground oats or barley, Alfata meat, fish meal, oyster shell, vitamin A, vitamin D and coccidiostat (which is given in form and at the level recommended by the manufacturer). The different percentage of each ingredient varies depending on the kind of feed. There could be the same percentage of a particular ingredient in different feeds as in the case of alfafa meal which constitutes 10% in all the feed types (Ubong, 2016).
1.2 Statement of the Problem
To maintain healthy fish, the feeds are kept fresh as much as possible at all times. The amounts of administered feeds are limited to the extent necessary to avoid wastage. Most fish feeds are prepared in dehydrated forms and because of this, there is the need to store them properly to avoid moisture uptake and damage by heat, in this way they can remain safe for a considerable period of time without losing their safeness and value. The feed should be stored in a suitable place where it will not be attacked by microorganisms, insects, rodents etc. Air tight storage of the various feeds is not advisable because offensive odour might result when there is prevention of outflow and inflow of air (Leonard, 1981). The condition of feed storage and handling could be a source of contamination. When feeds are un-hygienically handled and stored, there could be a build-up of microbial contaminants. This study was therefore aimed at isolation, identification, enzymatic potentials and antimicrobial susceptibility of bacterial associated to fish feed
1.3 Objectives of the Study
The main objective guiding this study is isolation, identification, enzymatic potentials and antimicrobial susceptibility of bacterial associated to fish feed.
While the specific objectives are:
1. To isolate and identify the bacterial associated with fish feed
2. To identify the enzymatic potential of bacteria associated with fish feed
3. Access the antimicrobial susceptibility of bacteria associated with fish feed.
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