UNIVERSITY FOR DEVELOPMENT STUDIES

FACULTY OF AGRICULTURE

THE PREVALENCE AND ANTIMICROBIAL RESISTANCE OF CAMPYLOBACTER

JEJUNI ON THE INTESTINES OF BEEF IN THE NORTHERN REGION

DORIS AFI SENAM BIBI (FST/0033/21)

NAKPAMBAL NILEMORCHA PATIENCE (FST/0082/20)

A PROJECT PROPOSAL SUBMITTED TO THE DEPARTMENT OF FOOD SCIENCE

AND TECHNOLOGY OF THE UNIVERSITY FOR DEVELOPMENT STUDIES IN

PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF A

BACHELOR OF SCIENCE IN FOOD SCIENCE

2024
Contents
1.0 INTRODUCTION....................................................................................................................3
1.1 BACKGROUND OF STUDY.......................................................................................................3
1.2 PROBLEM STATEMENT...........................................................................................................5
1.3 SIGNIFICANCE OF THE STUDY................................................................................................6
1.4 RESEARCH OBJECTIVES..........................................................................................................6
1.5 RESEARCH QUESTIONS..........................................................................................................7
1.6 MATERIALS AND METHODS...................................................................................................7
1.6.1 SAMPLE SIZE..................................................................................................................8
1.6.2 SAMPLE COLLECTION.....................................................................................................8
1.6.3 INOCULATION OF COLLECTED SAMPLES........................................................................9
1.6.4 ISOLATION AND IDENTIFICATION PROCEDURE..............................................................9
1.6.5 ANTIMICROBIAL SUSCEPTIBILITY TESTING.....................................................................9
1.7 QUESTIONNAIRE..................................................................................................................10
1.8 DATA ANALYSIS....................................................................................................................10
1.9 ETHICAL CONSIDERATIONS..................................................................................................10
References...........................................................................................................................................11
1.0 INTRODUCTION

1.1 BACKGROUND OF STUDY

Beef constitutes approximately 24% of domestic meat production and 13.6% of imported

meat products in Ghana, indicating a high patronage by consumers (Hughes et al., 2015). The

rich nutritive content of beef makes it susceptible to microbial contamination, since it

promotes growth and multiplication of microorganisms. Contaminated beefs are, thus,

rendered perishable, if not properly handled and preserved (Kebede & Getu, 2023; Yafetto et

al., 2019). Carcass contamination during slaughter is the major source of microbial

contamination. Carcass contamination practices include the use of unsterilized and

improperly cleaned knives and equipment, dressing of carcass on filthy slaughter floors and

hanging of meat in open places overnight (Olu-Taiwo et al., 2021). In most cases, personal

hygiene is highly compromised as people who are slaughtering animals rarely appear to wash

their body or hands and use their own clothes without protective clothing (Tanganyika et al.,

2017).

Microbial contamination of meat is a major cause of food poisoning and foodborne illnesses

worldwide (Wazir Shafi, 2021). An estimated 9.4 million illnesses are caused from foodborne

diseases by known microbes each year in the United States (Vieira, 2015). Bacterial

pathogens, including Campylobacter, Escherichia coli (E. coli) O157:H7, Staphylococcus

aureus, Salmonella, and Enterococci, are among the top five foodborne pathogens and

globally account for 230,000 deaths each year (Adjei et al., 2022; Ncoko & Jaja, 2020).

Moreover, protozoan parasites such as Toxoplasma gondii (T. gondii) and Cryptosporidium

parvum (C. parvum) are crucial foodborne pathogens associated with the intake of infected

raw or undercooked meat. These bacterial and protozoan pathogens are a public health
problem and negatively impact the economy in terms of loss of productivity, morbidity, and

healthcare cost (Adjei et al., 2022; Augendre et al., 2023).

As reported by the World Health Organization (W.H.O.), there are an estimated 96 million

cases of diarrheal illnesses worldwide, and campylobacter spp. Amount to one of the most

common causes of infectious gastroenteritis in people (Kirk et al., 2015). It is acknowledged

that the most frequent cause of human campylobacteriosis is exposure to tainted flesh. Due to

their presence in the gut flora of the majority of domestic beefs, campylobacter spp. are

transferred from the corpse to the poultry meat during the slaughter process (Igwaran &

Okoh, 2019). Human exposure to Campylobacter is known to be heightened by improper

handling and/or undercooking of beef (Chagneau et al., 2023). According to Dai et al.

(2020), Campylobacter sp. are the most often identified species in human cases; an

infectious dosage of these bacteria can be as low as a few hundred. Human symptoms include

fever, diarrhoea, malaise, and cramping in the abdomen (Nisar et al., 2017). According to

Nisar et al. (2017), the majority of Campylobacter infections in healthy individuals resolve

on their own and don't need antibiotic treatment. In cases of severe illness, macrolides and

fluoroquinolone antibiotics are regarded as first- and second-line treatments, respectively

(Deckert et al., 2013).

While studies in high-income nations frequently report on the prevalence of Campylobacter

spp. contamination of beef and beef products and the resistance patterns of isolates, there is

comparatively little data available for the African continent (Paintsil et al., 2023; Ramatla et

al., 2022). The lowest estimated incidence of Campylobacter spp. in beef is 9.6% and could

be as high as 90% among African nations (Parry-Hanson Kunadu et al., 2020). There have

been a few studies conducted in Ghana on animal-derived meals, and the prevalence has been

reported to be 11%–18% in poultry carcasses (Dekker et al., 2019; Parry-Hanson Kunadu et

al., 2020) and 13%–36% in carcasses from sheep, cattle, goats, and pigs (Karikari et al.,
2017b). These studies, however, have failed to evaluate the profiles of antimicrobial

resistance in beef; reports have indicated that 75% and 85% of beef meat is resistant to

ciprofloxacin and tetracycline, respectively (Dekker et al., 2019). However, because

antimicrobials are so widely available, antimicrobial resistance remains a serious concern to

public health in Ghana. This current study This current study will determine the prevalence of

antimicrobial resistance of Campylobacter jejuni on the intestines of beef in the Northern

region.

antimicrobials are so widely available, antimicrobial resistance remains a serious concern to

public health in Ghana.

1.2 PROBLEM STATEMENT

The presence of Campylobacter jejuni in the intestines of beef presents a major public health

concern due to its prospects to cause foodborne illness. Notwithstanding, being less studied

compared to poultry-related infections, emerging evidence suggests that beef may serve as a

reservoir for Campylobacter jejuni, leading to contamination during processing and handling

stages. According to the Ghana Food and Drink Report, there is a growing middle class and

expat population in Ghana, which has significantly increase in the consumption of beef. As a

result, restaurants and other food outlets are offering alternative processing options, like

medium and rare cooked beef, which could raise the risk of foodborne illnesses when

biological hazards are present (Duah, 2022). Therefore, research is necessary to determine the

safety and quality of beef sold to customers. The quality and safety of beef can be

significantly impacted by the actions of cattle farmers, butchers, and retailers who handle

meat before it is consumed (Akparibo et al., 2021; Hughes et al., 2015).

Campylobacter species are recognized as major causes of infectious gastroenteritis, with

exposure often linked to improperly handled or undercooked beef (Domingues et al., 2012;

Lim et al., 2017). While studies in high-income nations extensively report on Campylobacter
contamination in beef, there is a noticeable gap in such data for the African continent,

including Ghana (Carron et al., 2018; Asuming-Bediako et al., 2019). Limited studies

conducted in Ghana indicate Campylobacter prevalence in beef (11%–18%). Furthermore,

the existing knowledge on antimicrobial resistance in Campylobacter species in Ghana is

insufficient. Reports suggest high resistance levels in meat (75% and 85% to ciprofloxacin

and tetracycline, respectively), posing a significant public health concern (Dekker et al.,

2019). ). However, the prevalence and patterns of antimicrobial resistance in Campylobacter

jejuni on beef intestines in the Northern region particularly in these selected districts; Tolon,

Sagnarigu, Kumbungu and Tamale metropolis at large remain largely unexplored.

Addressing this gap is essential for understanding the local dynamics of antimicrobial

resistance and informing targeted interventions to enhance food safety and public health in

the region.

1.3 SIGNIFICANCE OF THE STUDY

This study leads to notable significance for public health and food safety in the Northern

region by unveiling the dynamics of antimicrobial resistance in Campylobacter jejuni found

in beef intestines. Understanding the prevalence and resistance patterns contributes vital

insights into potential health risks associated with the consumption of contaminated meat. By

identifying risk factors and genetic mechanisms, the research aims to inform targeted

interventions in local livestock management practices. The assessment of public health

implications addresses the broader context, shedding light on the potential transmission of

resistant strains from food sources to humans. This knowledge is crucial for developing

evidence-based strategies to mitigate the spread of antimicrobial resistance, safeguarding

both local communities and global efforts in combatting infectious diseases.

1.4 RESEARCH OBJECTIVES

 1. To assess the prevalence and patterns of antimicrobial resistance in Campylobacter

jejuni isolated from beef intestines in the Northern Region

2. To determine the overall prevalence of Campylobacter jejuni in beef intestines by

bacteria culture and isolation

3. Characterize the antimicrobial resistance profile of Campylobacter isolates through

antimicrobial susceptibility testing

4. Assess Risk Factors Contributing to Antimicrobial Resistance using a structured

survey

1.5 RESEARCH QUESTIONS

1. What is the overall prevalence of Campylobacter jejuni in beef intestines?

2. How is the antimicrobial resistance of Campylobacter isolates characterized?

3. What are the risk factors contributing to antimicrobial resistance in Campylobacter

isolates?

1.6 MATERIALS AND METHODS

The study will be conducted in some selected Districts and Tamale metropolis of the

Northern region of Ghana. It is located in the central part of the Region and shares boundaries

with the Sagnarigu District to the west and north, Mion District to the east, East Gonja to the

south and Central Gonja to the south-west (Fig. 1). The Metropolis has a total estimated land

size of 646.90180sqkm (GSS, 2014). Geographically, the Metropolis lies between latitude

9º16 and 9º 34 North and longitudes 0º 36 and 0º 57 West (GSS, 2014). The Ghana Statistical

Service’s 2021 report divulges that the Tamale metropolis has an estimated population of

374,744. This population is segregated into male and female populations of 185,051 (49.4%)

and 189,693 (50.6%) respectively (GSS, 2021).

Figure 1 Administrative Map of Tamale
Source: Ghana Statistical Service (2014)
1.6.1 Sample size
The research will be conducted in some selected districts and the tamale metropolis of the

Northern Region. The selected districts are: Tolon, Kumbungu, Sagnarigu and Tamale

metropolis at large. Tamale, the capital of the Region is the third largest city in

Ghana and located 600 km north of Accra, the capital of the country.

1.6.2 Sample collection

Beef samples will be obtained randomly from vendors, producers and processors in the study

area. All samples will be collected and stored using airtight containers and kept on ice pack to

minimize exposure to external conditions and potential contamination during transportation

to the laboratory. Collected samples will be legibly labelled.

1.6.3 Inoculation of collected samples
Culture media will be prepared according to the manufacturer’s instructions. Samples will be

inoculated on nutrients and Campylobacter blood free selective agar.

1.6.4 Isolation and identification procedure

Each swab will be streaked directly onto plates of Campylobacter Blood Free agar (Oxoid)

which will be supplemented with CCDA Selective Supplement (Oxoid) containing

cefeoperazone (32mg/l) and amphotericin B (10mg/l).

1.6.5 Antimicrobial susceptibility testing

Laboratory analysis of the collected beef samples will be carried out at the University for

Development Studies Spanish Laboratory. Antimicrobial susceptibility testing will be carried

out to measure the antimicrobial resistance activity of the Campylobacter strains to the

selected antimicrobial agents. This will be carried out in the Microbiology laboratory of the

University for Development Studies Spanish Laboratory. Accordingly, the Campylobacter

strains will be isolated from the beef samples using appropriate microbiological techniques.

Subsequently, a sub-culture of a pure culture of the isolated bacterial strains will be prepared

on a suitable agar medium. The Mueller-Hinton agar plates will later be inoculated with the

standard bacterial suspension using sterile swabs. Finally, the antibiotic discs for the selected

antibiotics will be inoculated on the agar plate and incubated at 42°C for 24-48 hours. The

zones of inhibition of the antibiotic discs will be measured. The following antibiotics will be

evaluated in this study for antimicrobial resistance of Campylobacter spp.: Tetracycline,

Chloramphenicol, Kanamycin, Erythromycin, Gentamicin and Ampicillin.

1.7 QUESTIONNAIRE
Additionally, a well-structured questionnaire will be used to obtain relevant information to

food safety awareness from retailers. The main areas of assessment will be beef processing,

storage, selling conditions, personal and environmental hygiene, and waste disposal as well

as hygienic handling of used equipment such as knives and water.

1.8 DATA ANALYSIS

Microsoft Excel will be used for data handling, organization, cleaning and coding.

Quantitative data will be analyzed using the Statistical Product for Service Solution (SPSS)

version 26. Categorical variables will be analyzed using percentages and frequencies.

Descriptive statistics will also be analyzed using measures of centrality (e.g., standard

deviation, mean, standard error, etc.). Fischer’s exact chi-square and one-way ANOVA will

be used to respectively determine any statistical differences between categorical and

continuous variables. Statistical significance will be set at 0.05.

1.9 ETHICAL CONSIDERATIONS

Ethical clearance will be obtained from the Research Ethics Committee of the University for

Development Studies prior to the start of the study. Permission and clearances will

subsequently be sought from any other relevant authority or bodies, if required, beforehand.

The informed autonomous consent of participants will be obtained before they partake in the

study. Data obtained from the study will be solely used for academic purposes. The identities

of the participants will be kept highly confidential and will not be divulged to a third party

without their permission

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