A RESEARCH PROPOSAL ON ANTIBIOGRAM OF CAMPYLOBACTER JEJUNI ISOLATED FROM FROZEN CHICKEN AND MEAT PRODUCTS

MADONNA UNIVERSITY,

ELELE, RIVERS STATE,

NIGERIA.

ANTIBIOGRAM OF CAMPYLOBACTER JEJUNI ISOLATED FROM FROZEN CHICKEN AND MEAT PRODUCTS SOLD IN SELECTED MARKETS IN OWERRI.

BY

AZAGBAEKWE OBIAJURU BENEDICT.

MB/12/069

A RESEARCH PROPOSAL SUBMITTED TO THE DEPARTMENT OF MICROBIOLOGY, FACULTY OF NATURAL SCIENCES, MADONNA UNIVERSITY IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF A BACHELOR OF SCIENCES (B.Sc.) IN MICROBIOLOGY.

SUPERVISIOR: DR WESLEY BRAIDE

MAY, 2016

CONTENTS

INTRODUCTION

AIM

OBJECTIVES

MATERIALS AND METHODS

STATISTICAL ANALYSIS

REFERENCE

  

INTRODUCTION

The availability, safety and portability of food materials are of global concern (WHO, 2013). The quality of food can be assessed by physical characteristics such as colour, flavour, taste, texture, etc. However, microbial quality of food to a large extent cannot be ascertained by mere physical means. Food substances that appear physically/organoleptically satisfactory may yet be highly contaminated with food borne pathogenic organisms. Routine check of food products for microbial quality is therefore highly required. Campylobacter jejuni is one of the food borne pathogens with a world leading impact as a causative or etiological agent of diarrheal disease known as Campylobacteriosis, being a leading cause of zoonotic food borne illness in industrialized countries and the second or third cause in developing countries (Fernandez, 2011; WHO, 2013). Transmission to human is usually by consumption of food substances of animal origin, handling of raw chicken and other poultry meat or offal and direct contact with animals or their exudates or faeces (Fernandez, 2011). Campylobacteriosis is often self-limiting and does not require antimicrobial treatment. However, in some cases such as septicaemia or other invasive forms of the disease, characterized by severe and prolonged enteritis as well as in immunocompromised or very young patients, antibacterial therapy may be needed. Macrolides (erythromycin) and quinolones, including fluoroquinolones (ciprofloxacin, nalidixic acid), are usually used in the treatment of Campylobacter infections (Wieczorek et al., 2012). 

Problem statement

Campylobacter jejuni is a leading cause of diarrhea called Campylobacteriosis, there is paucity of information about the pathogen in the Southeastern Nigeria. Report on increased resistance of Campylobacter sp to antibiotics calls for regular survey on its antibiogram and the need to screen for alternative sources of therapy. This project addresses the incidence of the pathogen in the south eastern Nigeria, its antibiogram and possible phytotherapy using Imo state as a case study.

AIM

The present study is aimed to determine the effect of antibiotics resistance pattern of Campylobacter jejuni in southeastern Nigeria. 

OBJECTIVES    

·        To study the effect of Campylobacter jejuni on frozen chicken and meat product.

·        To determine the risk of Campylobacter jejuni on frozen chicken and meat product.

·        To present a review of occupational exposure to Campylobacter jejuni

·        To determine antimicrobial susceptibility of Campylobacter jejuni 

Scope of the project

1.     Meat of different kinds (frozen chicken and beef) and from different locations in Owerri will be analyzed in this study and targeting up to 200 isolates of Campylobacter sp.

2.     The isolates identified as Campylobacter jejuni will be subjected to antimicrobial sensitivity testing using standard Gram negative and positive antibiotics discs from Oxoid to obtain antibiogram.

MATERIALS AND METHODS

Study Area

The study will be done primarily in a laboratory.

Description of sample

Chicken and beef are animal parts processed unhygienically in the market due to transportation and contaminated environment.

Sample collection

A total of 20 samples each of chicken, and beef will be obtained in Ekenuwa market in Ihiagwa, Imo State. Samples will be purchased directly from the butchers into a transparent polyethylene bag. They will be collected randomly and transported to the laboratory for microbiological analysis.

Isolation of test organisms                                                              

Pure cultures of different test organisms shall be obtained from meat samples, ten folds serial dilutions shall be made up to 10^-6 and approximately, 0.1ml aliquot of dilutions will be surface plated in duplicates on a suitable medium. The plates shall be incubated at 37^oC for 24hrs. Following incubation, the plates shall be examined and colonies characterized using standard methods.

Microbiological examination of samples

Washing, chopping and blending of sample will be carried out, extract the water into a Campy medium and will be incubated at 37^oC for 24-48hrs. The plates will be observed for the production of white colonies which characterizes the Campylobacter sp on Campy Agar.

Characterization and identification of bacterial isolates

Pure cultures of bacterial isolates shall be identified on the basis of their colonial morphology, microscope and biochemical characteristics. Isolates shall be identified with reference standard bacteriological manual.

Media to be used

Campylobacter Agar.

Mueller-Hinton agar

Antimicrobial Activity   

The testing of the bacterial cultures for the inhibitory effect of Standard Oxoid antibiotics on Campylobacter jejuni shall be performed using the disc diffusion method. Active cell suspensions of the test organisms standardized with equivalent of 0.5 McFarland shall be spread uniformly with the help of a sterile swab stick on Mueller-Hinton agar. Each experiment shall be done in triplicates and zone of inhibition measured in milliliter diameter shall be recorded and standard deviation calculated. Negative control shall be done using sterile distilled water in place of the Campylobacter jejuni. Plates shall be incubated at 37^oC for 24 hours before being examined for zone of inhibition

REFERENCE

Aarestrup, F.M. and Engberg, J. (2001). Antimicrobial resistance of thermophilic Campylobacter. Vet. Res.  32 (3-4): 311–321.

Abrahams, C.A., Agbodaze, D., Nakano, T., Afari, E.A. and Longmatey, H.E. (1990). Prevalence and antibiogram of Campylobacter jejuni in domestic animals in rural Ghana. Arch. Environ. Health 5(1):59-62.

Akwuobu, C. A.,Oboegbulem, S. I. and Ofukwu, R. A. (2010). Characterization and antibiogram of local isolates of Campylobacter species from chicken in Nsukka area, Southeast Nigeria. American-Eurasian Journal of Sustainable Agriculture 4(2): 117-121.

Angelovski, L., Sekulovski, P., Jankuloski, D., Ratkova, M., Prodanov, M., Kostova, S. (2011). Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolates from broiler flocks. Mac. Vet. Rev. 34 (1): 15 – 18.

Black, A.P., Kirk, M.D. and Millard, G. (2006). Campylobacter outbreak due to chicken consumption at an Australian Capital Territory restaurant. CDI 2006, 30: 373–377.

Wieczorek, K., Szewczyk, R. and Osek, J. (2012). Prevalence, antimicrobial resistance, and molecular characterization of Campylobacter jejuni and C. coli isolated from retail raw meat in Poland. Veterinarni Medicina 57 (6): 293–299