BACKGROUND: Unsafe meat handling in butcher shops exacerbates microbial contamination, contributing to widespread food-related infections, particularly in developing countries like Ethiopia. In Ethiopia, microbial contamination in the meat supply chain is pervasive due to limited knowledge, inadequate hygiene practices, insufficient training, poor enforcement of food safety standards, and the absence of advanced diagnostic tools and regular microbial testing. This study aimed to assess the microbial load of meat, its contact surfaces, and meat handling practices of butcher shop workers in Hawassa city, Ethiopia.
METHODS: A cross-sectional study was conducted from March to April 2023 among butcher shop workers in Hawassa city, Ethiopia. One hundred forty-four (144) swab samples were collected, and the microbial load was evaluated by serial dilution method and counted using standard procedures. A total of 52 workers were interviewed to assess meat handling practices. Descriptive statistics were used to analyze survey and laboratory data.
RESULTS: The findings of this study revealed that the levels of Total Plate Count, Enterobacteriaceae, and staphylococci from meat samples collected from the butchers were 6.14, 5.58, and 5.23, respectively. The levels of Total Plate Count, Enterobacteriaceae, and staphylococci in the swab samples collected from knives, chopping boards, and weighing boards were 5.66, 5.27, and 5.28; 5.61, 5.33, and 5.03; and 5.29, 4.97, and 4.91, respectively. In the butcher shops, 83% of the meat was exposed to flies without protection, and 64% of meat handling equipment was not disinfected.
CONCLUSIONS: The Total Plate Count, Enterobacteriaceae and staphylococci levels in meat samples and swab samples collected from knives, chopping boards, and weighing boards were above acceptable levels. These findings suggest the presence of potential pathogenic microorganisms, likely due to poor personal hygiene of the workers, inadequate cleaning and sterilization in the butcher shops, and cross-contamination from sources, including paper money handled, and skin contact.
Introduction
Meat is an animal tissue used as food commonly consumed as a source of protein in many parts of the world.1,2 However, it is also considered a high-risk product for food-borne diseases due to its favorable composition for microbial proliferation.3-5 Globally, nearly 1 in 10 people worldwide, or an estimated 600 million individuals, get sick from foodborne diseases each year, with 420 000 deaths resulting from these illnesses. Africa bears a significant burden of foodborne diseases, with an estimated 91 million cases and 137 000 deaths annually.6 The consumption of food of animal origin such as meat and fish, especially in their raw state, is generally regarded as a health risk unless safety principles are observed.7
Meat contamination can occur during various stages such as slaughter, transport, and purchase.2,8,9 When sold in butcher shops, further contamination can occur through contact with an unclean butcher shop environment, workers’ hands, and unhygienic equipment such as cutting tables, scales, and knives.10,11 Raw meat can harbor different potentially pathogenic microbes such as Enterobacteriaceae, staphylococci spp., Campylobacter spp., Yesinia Enterocolitica, Listeria monocytogenes, making the meat a potential risk to human health and spoilage.8,12-14
Eating raw or partially cooked meat is common in Ethiopia with a significant lack of food safety in butchers, which can be a potential source of pathogens that cause foodborne illness.9,15 Unhygienic meat handling practices, inadequate or weak food safety regulations, lack of financial resources to invest in safer equipment and working areas, and lack of training for food processors all lead to compromising meat quality in developing countries like Ethiopia.9,16,17 The production of high-quality food is achieved by implementing quality control measures by workers who are in contact with food handling practices. This includes having an appropriate level of education, having training in personal hygiene, and following good food safety practices.16 To prevent cross-contamination of food, it is essential to take all reasonable precautions. This involves using safe water for all activities, maintaining personal hygiene, and thoroughly cleaning handling equipment and the meat processing area.18
The meat supply chain in Ethiopia faces significant challenges due to a substantial knowledge gap regarding microbial contamination factors. This gap encompasses inadequate understanding and implementation of best hygiene and sanitation practices at various supply chain stages, from slaughterhouses to retail markets. Inadequate training and awareness among workers, combined with insufficient monitoring and enforcement of food safety standards, exacerbate the risk of contamination. Additionally, the absence of advanced diagnostic tools and regular microbial testing further impairs the ability to detect and control contamination early in the supply chain.19 This knowledge gap is compounded by limited research on specific microbial risks prevalent in Ethiopia. The deficiency in localized studies and tailored intervention strategies means that potential sources of contamination and their mitigation are not well understood or effectively managed. Consequently, the meat supply chain remains vulnerable to outbreaks of foodborne illnesses, understanding public health and economic stability in Ethiopia, including in cities like Hawassa.20 Addressing these knowledge gaps through targeted research, education, and policy initiatives is crucial for enhancing the safety and quality of meat products. Additionally, it is important to determine the microbial load of meat and its contact surfaces to prevent contamination. While limited studies have been conducted on the microbiological characteristics of meat, contact surfaces, and the meat handling practices of butcher shop employees in other parts of Ethiopia, this study specifically aimed to assess the microbial load of beef and its contact surfaces, as well as the meat handling practices of butcher shop workers in Hawassa City, Ethiopia.
Methods and Materials
Description of the study area
The study was conducted from March to April 2023 in Hawassa city, Sidama National Regional State, Ethiopia, which is located 273 km from Addis Ababa, the capital city of Ethiopia. The city is situated on the shores of Lake Hawassa in the Great Rift Valley and has a latitude and longitude of 7°3′N 38°28′E/7.050°N 38.467°E and an elevation of 1708 m (Figure 1). The mean annual rainfall and temperature are 961 mm and 26°C, respectively. The population projection for Ethiopia in 2024 is approximately 129.7 million people, reflecting a growth rate of about 2.6% per year. Specifically, Hawassa city’s population in 2024 is projected to be around 133 097.21 The city administration has an area of 157.2 km2, divided into 8 sub-cities. According to the Hawassa City Administration Trade Register Department, there are 168 registered butcher shops in all sub-cities that actively provide services and are supplied by 1 abattoir. All registered butcher shops in the city are actively giving services. All the butcher shops in the city source their meat from abattoirs.
Study design and period
A cross-sectional study design was conducted from March to April 2023 to address the objectives of this study. Meat samples and swab samples were collected from knives, chopping boards, and weighing boards for laboratory analysis. In addition, the study included a questionnaire and observational survey to assess meat handling practices, the hygiene conditions of equipment, and the cleanliness of butcher shops.
Source and study population
The source population for this study was all butcher shops, workers from these butcher shops, and the equipment used in Hawassa city butcher shops. The study population included selected butcher workers and equipment in the selected butcher shops in Hawass city, Ethiopia. Meat handlers actively working in the selected butcher shop during the study period were included. Meat handlers who were seriously ill and unable to communicate during data collection were excluded from the study.
Sample size determination and sampling procedure
The sample size (n) was determined based on the total number of butcher shops in Hawassa city (N = 168) using a formula of a known population as given below.22
Where e = acceptable error (0.05), Z = standard variate at a 95% level of confidence (1.96) and SD = standard deviation of the sample mean obtained by the following formula.23
Where and σ = standard deviation of the population.
By considering the availability of resources the researchers adjusted the sample size using the formula cited in Nundy et al.24
Where nadj = adjusted sample size, N = the finite population size, and n = the sample size calculated in the equation.
Sample collection
A total of 144 swab samples were collected aseptically from 36 selected butcher shops to determine the microbial load. Thirty-six swab samples were collected from meat, and 108 (3 × 36) were collected from knives, cutting boards and weighing boards using sterile cotton swabs. Cotton-tipped swabs were soaked in a tube containing 10 ml of 0.1% peptone water solution and rubbed on the 50 cm2 area25 horizontally and vertically for 5 to 10 seconds then placed separately in sterile containers for microbiological analysis. All samples were labeled with necessary information, including the sampling date, sample code, and type. Subsequently, the samples were kept in an icebox and immediately transported to the Department of Environmental Health Laboratory, Hawassa University, for microbial analysis. The samples were stored in the laboratory at 4°C (refrigeration temperature) and processed within 24 hours.
Sample preparation
Twenty-five grams (25 g) of the meat were weighed and transferred to a stomacher bag aseptically. The samples were placed in test tubes containing 9 ml of peptone water and mixed for 2 minutes by vortexing. After that, further serial dilutions were performed using sterile peptone water. Each tube containing samples (10 ml of the 0.1% saline water) was vortexed to ensure a mixture of the samples. To prepare the first 10-fold dilution (10−1), 1 ml of the homogenized sample, which included meat or swabs, was transferred into 9 ml of the diluent. This process was repeated by transferring 1 ml from the 10−1dilution into another 9 ml of diluent to create a 10−2 dilution, and again by transferring 1 ml from the 10−2 dilution into a fresh 9 ml of diluent to prepare the 10−3 dilution. From each of these 3 dilutions (10−1, 10−2, and 10−3), a 0.1 ml aliquot was taken and plated onto various media types to culture bacteria for microbial counts. The samples were plated in triplicate for each dilution, resulting in 3 plates per dilution, and a total of 9 plates across all dilutions.18,26 Average counts from the three 10-fold dilutions were reported as an estimation of the microbial load.
Total Plate Count
A series of sample dilutions were prepared, typically involving serial dilutions for various concentrations. Each dilution (0.1 ml) was pipetted onto sterile Petri dishes. Molten Plate Count agar, cooled at approximately 45°C, then added to the dishes. The mixture was gently swirled to ensure even distribution and allowed to solidify at room temperature. Once solidified, the plates were inverted to prevent condensation from affecting the colonies and incubated at 35°C to 37°C for 24 to 48 hours. After incubation, colonies on the plates were counted using a colony counter.27
Staphylococci count
For culturing and identifying staphylococci on Mannitol Salt Aga, begin by preparing serial dilutions of the sample. Each (0.1 ml) dilution was spread onto the surface of pre-poured and dried Mannitol Salt Agar plates using a sterile glass rod and then incubated at 35°C to 37°C for 24 to 48 hours. After incubation, yellow colonies, which indicate the presence of staphylococci due to mannitol fermentation were counted and recorded as staphylococci using a colony counter.27
Enterobacteriaceae count
Each dilution (0.1 ml) was spread onto the surface of pre-poured and dried MacConkey Agar plates using a sterile glass rod. The samples were incubated at 35°C to 37°C for 24 to 48 hours. After incubation, all reddish purple or pink colonies, which indicate the presence of Enterobacteriaceae were counted and recorded using a colony counter.27
Determination of microbial load
Appropriate plates containing distinct microbial colonies were selected and counted using a colony. The microbial load was determined using the standard formula as follows.28
where N = total number of bacteria (CFU) per milliliter of the sample, n = average number of bacterial colonies, from different dilutions in a Petri dish that contained 30 to 300 colonies, s = volume of sample for plating, and d = dilution factor of the specimen/food sample.
Questionnaire and observational survey
The survey was conducted using questionnaires and visual observations. Semi-structured questionnaires were prepared and filled out by meat handlers in the butcher shops to assess meat handling practices. In total, 52 meat handlers from 36 selected butcher shops were interviewed about their meat handling practices. The number of workers interviewed varied: some butcher shops had 2 workers interviewed, while others had only one, depending on their availability during data collection. The questionnaire addresses sociodemographic characteristics and training on meat handling hygiene. The hygiene and cleanliness of the butcher shops were assessed by direct observation using a checklist. The checklist consists of items to check whether the butchers are properly washing their hands, wearing appropriate overcoats, cleaning the overcoats, wearing hair cover, wearing masks, wearing jewelry, handling money experience, checking the presence of skin rash, keeping fingernails short, presence of standing water with soap, and others. Additionally, the layout of the butcher shop, general cleanliness, the presence of standing water with soap, and the way meat was placed in the shop were directly observed using the checklist.
Data quality control
Training was provided to data collectors on the objective of the study, its importance, and confidentiality of information. Data collection was supervised to ensure the completeness and reliability of the gathered data throughout the data collection process. All media used for microbiological analysis were checked for expiration dates and prepared and sterilized according to their respective manufacturer’s instructions. Composite swabs were used on both the eat and its contact surfaces. Trained experts conducted the microbiological analysis of these samples.
Statistical analysis
Data were analyzed using Statistical Package for Social Sciences (SPSS) version 20.29 Descriptive statistics such as mean, standard deviation, frequency and percentage were used for presentation and summarization of data. All microbial counts were converted to log10 CFU/cm2 values and reported as means ± standard deviation. Descriptive statistics were used for survey and laboratory data while t-tests and One-Way Analysis of Variance were used to assess the mean significance (P < .05) in microbial loads from the sample groups. Further analysis of the post-hoc multiple comparison test was computed to assess where the significant differences existed.
Ethics approval and consent to participate
Ethical approval and clearance were obtained from the Institutional Review Board of Hawassa University, College of Medicine and Health Sciences and a support letter was obtained from the Hawassa city Administration Health Department. The study objective was explained to participant students and the participation was voluntary and confidential. A written and signed informed consent form was used to ensure willingness to participate in the study and the confidentiality of the data given. On top of that, informed consent was also obtained from the owners of the butcher shops. Written consent was obtained from owners/managers and workers of the butcher shops after assuring the aim of the study, data collection procedure, confidentiality issue of the obtained information, the rights of participants, and the risks and benefits of participating in the study.
Results
Sociodemographic information on butcher shop workers
In the present study, a total of 52 workers were interviewed. More than half (64%) of them attended only primary school, while 25% attended secondary school, 8% above secondary school, and 48% were illiterate. Regarding work experience, they had a minimum of 1 year and a maximum of 13 years of experience with a relatively higher number of workers (42%) having 5 to 10 years of work experience. In terms of meat safety training and health certificates, only 42% of meat handlers took training, and 38% of the workers had no health certificate. Most respondents (90%) reported that they were supervised by health professionals about their practice while the remaining 10% were never supervised (Table 1).
Table 1.
Sociodemographic information of butcher shop workers in Hawassa city, Ethiopia.
Based on visual assessment, only 27% of meat handlers wore overcoats (with visible dirt). Similarly, 69% and 94 of them did not wear hair cover and masks respectively. More than half (62%) of the workers handled money while processing meat, and 13% of the workers had visible skin lesions. During the study period, most butcher shops (85%) had no standing water even if 77% of the workers reported that they washed their hands properly (Table 2).
Table 2.
Meat handling practices related survey in butcher shops in Hawassa city, Ethiopia.
Based on the observation, majority of the butcher shops’ equipment (77%) was not sterilized. The building premises of most of the butcher shops were not smooth (concrete), waterproof and light in color, and dirty like spider webs and dust were observed on the wall and hook. About 86% of shops chop and display meat on tables without cover, making it easily exposed to flies because they lack proper fly-controlling mechanisms. Regarding the handling of the equipment, 81% were found to be in poor hygienic condition. Debris and unusable meat pieces after trimming were not well separated but left around the chopping board, exposing them to flies. Approximately 77%, 63%, and 71% of butcher shop equipment were not sterilized, disinfected and had no set cleaning schedule, respectively (Table 2).
Microbial loads of meat and contact surfaces
The microbiological analysis of meat samples has indicated the mean microbial count was 6.14, 5.58, and 5.23 log10 CFU/cm2 for Total Plate Count, Enterobacteriaceae and staphylococci, respectively. The mean microbial count from all contact surfaces were 5.52, 5.19, and 5.08 log10 CFU/cm2 for Total Plate Count, Enterobacteriaceae, and staphylococci respectively. There was a significant difference in the load of each microbial group tested in meat swabs and samples collected from the meat contact surfaces (Table 3).
Table 3.
Microbial load (log10 CFU/cm2) of meat and contact surfaces of butcher shops in Hawassa city, Ethiopia.
The mean Total Plate Count from knives, chopping boards, and weighing boards were 5.66, 5.61, and 5.29 log10 CFU/cm2, respectively. For Enterobacteriaceae and staphylococci, the mean load from knives, chopping boards, and weighing boards were 5.27, 5.33, and 4.97 log10 CFU/cm2, and 5.28, 5.03, and 4.91 log10 CFU/cm2, respectively. For individual sample points, a lower mean was recorded from the weighing boards, and there was a significant mean difference among sample groups for all microbial groups. Based on the post-hoc test, the significant difference in the mean loads of Total Plate Count and Enterobacteriaceae was found on weighing boards while the mean load of staphylococci was found on knives (Table 3).
Discussion
The present study has revealed that most of the respondents (64%) had only completed primary school. This educational background has significant implications for their meat handling practices. Similar findings were reported in a study conducted in Debre Berhan, Ethiopia, where 68% of respondents had only elementary school.26 These results highlight the need for targeted food safety programs to improve meat handling practices among individuals with limited formal education. In terms of meat safety training and personal hygiene, more than half of the respondents (56%) had not received any training. Previous studies found that most meat processors had not received training on meat handling and personal hygiene.4,30 Educating food handlers on basic principles of personal hygiene and food hygiene plays a crucial role in ensuring the delivery of safe food products to consumers.18
The present study revealed that workers have varying levels of meat processing experience, ranging from 1 to 13 years. Notably, the literature indicates that meat handlers with longer work experience tend to exhibit better food hygiene practices.16,30 These insights underscore the importance of experience in promoting safe meat handling practices. In this study, most workers (58%) reported not receiving any training on meat safety handling. The lack of training is common in developing countries, where butcher shop workers often neglect hygiene practices in meat processing, contributing to microbial contamination.18 This finding emphasizes the need for comprehensive training programs to enhance meat safety training to enhance meat handling practices, thereby reducing the risk of contamination.
According to the observations, only 27% of meat handlers who wore white overcoats had clean overcoats, and more than half (69%) did not wear a hair cover. This finding is supported by the study conducted in Addis Ababa city in which 75% of the respondents did not wear a hair cover.31 In addition, a study conducted in Jigjiga city found that only 38% of workers wore a clean overcoat17 and in the study in Mekelle city, 63% of meat workers were not wearing a hairnet.4 In this study, almost all workers (94%) did not wear a mask, which is consistent with a previous study that noted 99% of the respondents were not wearing masks.17
The present study found that more than half (62%) of meat sellers handled money (without a cashier) when serving meat. The study conducted in Adama town, Ethiopia, reported that 57% of meat sellers handled money during meat processing.30 However, it contrasts with the study done in Mekelle city which showed that 92% of workers handled money when processing meat,30 and the study in Nairobi and Isiolo counties found that more than 90% of respondents handled money.32
In developing countries, most transactions are conducted with paper money, which may be contaminated with many microorganisms.33 This study also revealed that 87% of the workers in the butcher shops had a skin rash. This finding is consistent with the previous studies which reported the presence of skin rashes among workers in butcher shops.16,31 Skin rashes among butcher shop workers not only raise health concerns due to potential discomfort and complications but also pose risks to hygiene and food safety as they can serve as entry points for pathogens, including the risk of meat contamination. To mitigate these issues, it is crucial to implement measures such as providing personal protective equipment and promoting good hygiene practices among workers.
This study found that 81% of meat contact surfaces and carcasses were unprotected and easily exposed to flies in 77% of butcher shops, where equipment was unsterilized, indicating unhygienic conditions. In addition, the study revealed that 63% of the butcher shop equipment was disinfected, and 72% did not have a cleaning schedule. This finding is supported by a study conducted on butcher shops in Gondor town, which reported that the hygiene condition was poor, with 77% and 83% of the meat contact surfaces and the carcasses unprotected and easily exposed to flies.11 The finding of this study is supported by previous findings, which showed that 79% of meat shop respondents did not use disinfectant to clean utensils,17 and that 100% of meat shops did not disinfect their shops and 95% did not sterilize their knives and other equipments.12
Regular cleaning of equipment in butcher shops and maintaining personal hygiene are essential parts of good hygiene practices. All places, surfaces and equipment that come into contact with meat must be washed and disinfected before and after daily work.34 The floor of the work area should be waterproof, light color, smooth, easily washable and cleanable.35 However, in the present study, most butcher shops failed to comply with these principles. Therefore, there is a possible risk of cross-contamination during meat processing that can lead to health risks to consumers.
According to the European Safety Authority, the limit for Total Plate Count should be less than 5 log10 CFU/cm2, Enterobacteriaceae less than 2 log10 CFU/cm2, and staphylococci less than 2 log10 CFU/ cm2 for beef carcasses.28 The findings of this study revealed that the levels of Total Plate Count, Enterobacteriaceae, and staphylococci from meat samples collected from the butchers were 6.14, 5.58, and 5.23, respectively, which were above the acceptable level. The levels of Total Plate Count, Enterobacteriaceae, and staphylococci in the swab samples collected from knives, chopping boards, and weighing boards were 5.66, 5.27, and 5.28; 5.61, 5.33, and 5.03; and 5.29, 4.97, and 4.91, respectively. These findings also indicate that the microbial loads of each microbe tested in swab samples collected from knives, chopping boards, and weighing boards were above acceptable levels. The higher microbial loads from meat samples and the contact surfaces indicate the potential presence of different pathogenic microorganisms. This could be attributed to unsafe meat handling practices, such as poor personal hygiene of the workers, inadequate cleaning, or absence of sterilization in the butcher shops, as well as handling of paper money and cross-contamination from skin and utilities in the study area.
Conclusions and Recommendations
The Total Plate Count, Enterobacteriaceae and staphylococci levels in meat samples and swab samples collected from knives, chopping boards, and balance boards were above acceptable levels. These findings indicate the potential presence of many pathogenic microorganisms which can lead to low-quality meat and serve as a potential source of foodborne infections. The study revealed that butcher shops with poor hygiene practices and facilities had significantly higher microbial loads on meat and contact surfaces. To address these issues, local authorities including Hawassa City Municipality, Sidama Region Public Health Institute, Ethiopian Food and Drug Authority, and Hawassa City Administration Trade Register Department focus on the implementation of hygiene principles to ensure the meat is safe for the consumers. Efforts should be made to enhance public awareness by introducing meat safety training courses, effective inspection procedures, and disseminating information.
Limitations of the Study
The microbes selected for assessing microbial contamination in the meat supply chain provide valuable information about general microbial load and hygiene conditions. However, it doesn’t cover all potential pathogens that could pose significant health risks. This limitation highlights the need for a more comprehensive selection of microbes to enhance the robustness and comprehension of the study.
Acknowledgements
The authors acknowledge Hawassa University, College of Medicine and Health Sciences for the provision of ethical clearance. The authors are also grateful to the Hawassa City Administration Trade Register and Health Departments, butcher shop managers, study participants, and data collectors for their kind cooperation during data collection.
This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License ( https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages ( https://us.sagepub.com/en-us/nam/open-access-at-sage).
Author Contributions
ZK, BKO, and DD contributed to the conception and design of the study, conducted the study, analyzed and interpreted the data, and drafted the manuscript. AE participated in reviewing the manuscript. All authors read the draft manuscript, improved and approved the final manuscript.
Availability of Data and Materials
Based upon reasonable request to the correspondent author, supporting data and material of this study are readily available.
