RESISTANCE PROFILE OF LACTIC ACID BACTERIA ISOLATED FROM SOME SELECTED ANIMAL FAECES

ABSTRACT

This study evaluated the antibiotic resistance profile of lactic acid bacteria isolated from some selected animal faeces. A total of Twelve (12) faecal samples were collected from some pigs and poultry in Umudike, Ikwuano local government of Abia state and each sample was put into different polyethene bags. All samples were cultured using the spread plate method on MRS agar and incubated. A total of 14 Lactic acid bacteria comprising of 3 different species were isolated from the poultry faeces. The details of these lactic acid bacterial isolate comprises of  Lactobacillus  fermentum, Lactococcus  lactis and Lactobacillus reuteri. it was observed that Lactobacillus fermentum is the most frequently occurring isolates with a percentage occurrence of 8(57.1%), followed by Lactobacillus reuteri with a percentage occurrence of 4(28.6%), then Lactococcus lactis with a percentage occurrence of 2(14.3%). The antibiotic profile of the isolates reviewed that LAB regardless of species obtained from faecal samples showed high sensitivity to Levofloxacin (92.9%) followed by Ofloxacin (85.7%), Ciprofloxacin (78.6%) then, Gentamicin (71.4%) whereas the resistance profile of Lactic acid bacteria isolates to antibiotics reviewed that the Lactic acid bacteria isolates exhibited high rate of resistance to Augmentin and Ceftaxidime with percentage resistance of (100%), this is followed by Cefotaxime (92.9%), Erythromycin (85.7%), Cetriaxone (71.4%) and Azithromycin (71.4%). These results indicate that LAB may develop resistance against antibiotic that may result from the horizontal transferring of resistant gene to other microflora in the gut. The development of resistance to antibiotic can be attributed to long term usage of antibiotic as therapeutics and growth promoter. Thus, it is important and essential to advise the farmer a proper way of antibiotic use as therapeutic and growth promoter agents. Additionally, supplementation of unknown source of probiotic as feed additive needs to be monitored closely in animal feeding.

TABLE OF CONTENTS

Title                                                                                                  Page

Title Page                                                                                                                                 i

Certification                                                                                                                           iii

Dedication                                                                                                                              iv

Acknowledgement                                                                                                                  v

Table of Contents                                                                                                                   vi

List of Tables                                                                                                                          viii

Abstract                                                                                                                                  ix

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Aims and Objectives                                                                                                  2

CHAPTER TWO

2.0       Literature Review                                                                                                       3

2.1       Lactic Acid Bacteria                                                                                                   3

2.1.1    Classification and physiology of LAB                                                                       4

2.1.2    Antimicrobial mechanisms of LAB                                                                            5

2.1.2.1 Bacteriocins                                                                                                                6

2.1.3    LAB as food preservatives                                                                                         7

2.1.4    Application of the Inhibitory Activity of LAB to Pathogens                                     7

2.1.5    Safety and regulation of lactic acid bacteria                                                              9

2.2       Prevalence of Antibiotic Resistance in Lactic Acid Bacteria Isolated From the

Faeces of Broiler Chicken                                                                                          11

2.3       Characterization of Lactic Acid Bacteria Isolated From Poultry Farms                                     13

2.4       Antibiotic Resistance of Lactic Acid Bacteria from Canine Faeces                                    14

CHAPTER THREE

3.0       Materials and Methods                                                                                               16

3.1       Collection of Samples                                                                                                16

3.2       Sterilization of Materials                                                                                            16

3.3       Normal Saline Preparation                                                                                         16

3.4       Media Preparation                                                                                                      17

3.5       Isolation of Lactic Acid Bacteria                                                                               17

3.6       Sub-Culturing                                                                                                             17

3.7       Characterization and Identification of Lactic Acid Bacterial Isolates                        18

3.7.1    Gram Staining Techniques                                                                                         18

3.7.2    Biochemical Test                                                                                                        18

3.7.2.1 Motility test                                                                                                                18

3.7.2.2 Catalase test                                                                                                                19

3.7.2.3 Coagulase test                                                                                                             19

3.7.2.4 Methyl red test                                                                                                            19

3.7.2.5 Voges-proskaeur test                                                                                                  19

3.7.2.6 Indole test                                                                                                                   20

3.7.2.7 Citrate test                                                                                                                   20

3.7.2.8 Oxidase test                                                                                                                20

3.8       Antibiotic Resistance Testing                                                                                     21

CHAPTER FOUR

4.0       Results                                                                                                                        22

4.1       Total Lactic acid Bacteria counts of pig and bird fecal samples                              22

4.2       Morphological and Biochemical Characterization of Isolated Lactic acid

Bacteria                                                                                                                      22

4.3       Percentage Occurrence of the Lactic Acid Bacteria Isolates                                     22

4.4       Distribution of Lactic Acid Bacteria from the Samples                                             22

4.5       Antibiotics Profile of the Lactic Acid Bacterial Isolates                                           23

4.6       Resistance Profile of the Isolates                                                                               23

CHAPTER FIVE

5.0       Discussion Conclusion and Recommendation                                                           31

5.1       Discussion                                                                                                                   31

5.2       Conclusion                                                                                                                  33

5.3       Recommendation                                                                                                       33

References                                                                                                                  34

LIST OF TABLES

CHAPTER ONE

1.0       INTRODUCTION

Antibiotic are normally used to treat microbial diseases since 50 years ago. However, excessive use of antibiotic may lead to the development of antibiotic resistance in pathogenic bacteria. The report of antibiotic resistance is significantly increased due to the overuse and misuse of antibiotics, which has created an enormous selective pressure on the recessive bacteria. Antibiotic resistant bacteria have the ability to resist toward the actions of naturally occurring or synthetically produced compounds inimical to their survival (WHO, 2007).

Scott (2002) reported the identical resistance gene present in bacterial species isolated from different hosts. Antibiotic resistant may acquires through the in-vivo gene transfer between normal flora of gastrointestinal and antibiotic resistant pathogenic bacteria (Schjørring and Krogfelt, 2011). In fact, the antibiotic resistant pathogenic bacteria pose a great potential threat to human health, especially when the immunity system is not functioning well. Most of the developed countries have prohibited the use of antibiotic as animal growth promoter. Thus, various alternatives have been explored to replace antibiotic as growth promoter. One of the most common and popular alternatives is the application of probiotic as growth promoter in livestock animals.

In addition, the probiotic effects of postbiotic metabolites produced by probiotic strains have been shown in various animal species, such as rats, broilers, laying hens and post weaning piglets, suggesting they havegreat potential to be used as growth promoter for livestock animals.

Probiotic comprises of beneficial bacteria such as lactic acid bacteria (LAB). LAB is a group of gram-positive anaerobic bacteria which produce predominantly lactic acid from carbohydrate fermentation. Many farmers use probiotic extensively and those bacteria have potential to serve as a host of antibiotic resistance genes with a risk of transferring those genes into many LAB and other pathogens (Gueimonde et al., 2013).

The antibiotic resistant LAB has been detected by using DNA sequences which is responsible for antibiotic resistance traits. Egervärn et al. (2009) reported the emergence of antibiotic resistant in Lactobacillus reuteri and Lactobacillus plantarum. Moreover, Lactobacilli, Pediococci and Leuconostoc spp. have been reported to be highly resistant to vancomycin and some Lactobacilli have high resistance to bacitracin, cefoxitin, ciprofloxacin, fusidic acid, streptomycin, sulphadiazine, teicoplanin and vancomycin(Danielsen and Wind, 2003). Most of the reported LAB that resistant to antibiotics was isolated from food sources. These include the most commonly used probiotic species such as Lactobacillus casei, Lactobacillus acidophilus, L. reuteri, or Lactobacillus rhamnosus, among others, or the yogurt starter bacteria Lactobacillus delbrueckii(Ammoret al., 2008).

1.1       AIM AND OBJECTIVES

The aimof this study is to investigate the antibiotic resistance profile of Lactic Acid Bacteriaisolated from some selected animal feaces.

The objectives are;

1.     To isolate Lactic Acid Bacteria isolates from some selected animal feaces.

2.     To determine the percentage frequency distribution of Lactic Acid Bacteria isolates from some selected animal samples.

3.     To determine the antibiotic resistance profile of the Lactic Acid Bacterial isolates.

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