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Detections of antimicrobial resistance phenotypes and extended-spectrum beta-lactamase (ESBL)- producing Salmonella spps and Escherichia coli O157:H7 in raw vegetables and fruits from open markets in Jimma town, Ethiopia and evaluation of hygiene and handling practices of vendors

One Health Outlook volume 7, Article number: 2 (2025) Cite this article

Abstract

Objectives

Despite of the health benefits of consumption of fresh vegetables and fruits, this product could be associated with food-borne bacterial pathogens, including infections with antibiotic-resistant strains especially in developing countries due to limited in knowledge, and hygienic practices. This study was conducted to provide evidence data on the rates of Salmonella spp. and E. coli O157:H7 contamination, the antimicrobial resistance profile, and extended-spectrum β-lactamase (ESBL)-producing strains in fresh vegetables and fruits sold in open-air markets at Jimma town, southwest Ethiopia. In addition, this study provided data on the hygiene and handling practices of vendors, which can help as impute to improve food safety and safeguard public health. A total of 242 salad samples were collected from three different kebeles and examined for the presence of Salmonella spp. and E. coli O157:H7 in the microbiology laboratory of Jimma University by using conventional microbiological techniques.

Results

Out of 242 samples tested, 12.8% (31/242) were contaminated with Salmonella spp. and E. coli O157. Of these, Salmonella spp. was detected in 10.7% (26/242) of the tested samples, whereas Escherichia coli O157:H7 was found in 2.1% (5/242) of samples. Fifty-three-point-8% of Salmonella spp. were resistant to ampicillin, 42.3% to co-trimoxazole, 46.2% to tetracycline, and 26.9% resistance was observed against each of ceftriaxone and cefotaxime. 40% of E. coli O157:H7 isolates were resistant against ampicillin, amoxicillin-clavulanic acid, and co-trimoxazole. Only one isolate was resistant to ceftriaxone and cefotaxime, and no resistance was observed against ceftazidime, gentamicin, ciprofloxacin, chloramphenicol, and meropenem. Four Salmonella spp. and one E. coli O157:H7 isolate with a total of 5/31 (16.1%) isolates were confirmed as the ESBL producers. Multidrug resistance (MDR) was detected in 23.1% of Salmonella and 20.0% of E. coli O157:H7. Hygienic and handling practices of vendors were poor, which could contribute to contamination of vegetables and fruits in the area.

Conclusions

Contamination of fresh salad vegetables with pathogenic bacteria could be a food safety concern in the study area. Hence, this finding suggests the need for attention by the concerned bodies to prevent the emergence and transmission of food-borne pathogens and antimicrobial-resistant strains through these food items in the study area.

Background

The World Health Organization (WHO) and Food and Agriculture Organization (FAO) promote the daily consumption of fruits and vegetables as part of a healthy diet due to their high nutritional value [1]. On the other hand, the increased consumption of fruits and vegetables in recent years has been found to be accompanied by an increase in the number of human infections and outbreaks, as these can serve as reservoirs for pathogens or opportunistic pathogens. Antimicrobial resistance (AMR) can be transferred to humans through food, and fresh produce can be an ideal vector as it is often consumed raw or minimally processed [2, 3].

The microbiological contamination of foods of plant origin (e.g., fruits, vegetables, lettuce) that are consumed raw or undercooked can be contaminated with spoilage or pathogenic bacteria at any stage of the food chain, from primary production to consumption, and is responsible for foodborne illnesses, especially in developing countries, including outbreaks of disease caused by antimicrobial-resistant bacteria [4, 5]. For instance, the WHO says unsafe food causes 600 million cases of foodborne diseases worldwide and 420, 000 deaths, and some 30% of foodborne deaths occur among children under five years of age [6]. The prevalence of Salmonella on lettuce in developing countries was 6.4%, and in developed countries it was 2.8%, and the prevalence of Escherichia coli O157:H7 on lettuce was 4.1% [7]. Microbial contamination of fruits and vegetables regularly occurs in plantation fields through contact with soil, dust, contaminated irrigation water, and the use of raw sewage or manure fertilizers [8, 9]. The process of transporting these food products from the farm to households and to vending sites also contributes to the contamination of these fruits and vegetables, thus posing a serious problem in food safety [8, 9].

In Ethiopia, as it is a low- income country, facing multiple food safety challenges due to a lack of infrastructure and basic pre-requisites for food safety, such as clean water and environment, washing facilities, which compounded by limited implementation of food safety regulations [10]. Among some of clinically important foodborne pathogenic bacteria such as Salmonella spp. and E. coli O157:H7, showing high levels of resistance to most of the antibiotics prescribed in the country [10, 11]. For instance, the prevalence of Salmonella spp. detected in vegetables ranged from 0 to 10% [10], and the average prevalence of E. coli O157:H7 and Salmonella-related diarrhea in children and adults was 15.3% and 7.2%, respectively, and some of the factors associated with the infection were consumption of undercooked meat, raw vegetables, and/or fruit juice [12, 13].

To control infectious diseases, chemotherapy alone will not be enough; a concerted effort to limit and eliminate possible sources of infection is needed [14]. To address this, the periodic detection of infectious pathogens in food and related sources is a priority goal. Generating microbiological data of microbial contamination of vegetables and fruits is imperative to understand the burdens, guide the actions to prevent human health risks and other adverse consequences, and ensure produce safety on a sustainable scale. Therefore, this study was carried out to investigate the level of Salmonella spp. and E. coli O157:H7 contamination, antimicrobial resistance profiles, and extended-spectrum beta-lactamase (ESBL)-producing strains in fresh vegetables and fruits sold in open-air markets at Jimma town, southwest Ethiopia. Additionally, this study was conducted to provide evidences on the hygienic conditions and handling practices of vegetable and fruit vendors in a town. This preliminary data might help local authorities to achieve a better understanding of what may happen during the vegetables and fruits distribution from a farm grower to the market and to take further decisions to help both farmers and sellers for further consumer protection from developing diseases associated with the consumption of contaminated products with drug-resistant pathogens.

Materials and methods

Study design, area, and period

A descriptive cross-sectional study was carried out in three peri-urban areas (Hora Gibe, Bore, and Jiren markets) of Jimma town from first July to September 30, 2021. Jimma town is about 345 kilometers away from the capital city, Addis Ababa, in the southwest direction. The town is located at 7º 40’ North latitude and 36 º 5’ East longitudes, and the climate condition is relatively cool tropical monsoon climate with an average altitude of about 1780 m above sea level, a mean annual maximum temperature of 30 °C, and a mean annual minimum temperature of 14 °C. The annual rainfall ranges from 1138 mm to 1,690 mm. The fresh vegetables and fruits sold in these markets were brought from different agricultural areas found in the town and surrounding rural areas in a zone. As a result, diverse types of fruits and vegetables are frequently utilized as sources of food in the town.

Data collection and processing

Data on socio-demographic, hygienic and handling practices of vendors

Face-to-face interviews and visual inspections of local vendors were made using structured questionnaire and checklist to collect data on predisposing factors for bacterial contamination of vegetables and fruits in the study areas. Socio-demographic characteristics of vendors as well as the hygienic and handling-related factors associated with bacterial contamination of fresh fruit and vegetables were collected from randomly selected vendors of over 15 years of age, and from those the required samples (the selected fruit and vegetables) were obtained. In addition, hygienic and handling practices of vendors and the characteristics of vending sites were assessed using observational checklists and closed-ended questionnaires. The required number of samples were collected for around 12 weeks, from first July to September 30, 2021.

Data related to food safety knowledge

The street vendor awareness on contamination of fresh fruit and vegetables with pathogens, on personal and environmental hygiene, and on proper handling and cleaning was also assessed by face-to-face interviews and visual inspections of local vendors using a structured questionnaire. The questions have three possible answers: “yes”, “no” and “do not know”. Each “yes” answer was awarded one point, with the other answers being awarded 0 points. The total questions have a maximum of 100 points, where a score of less than 50 was considered to indicate a low level of food safety knowledge, 50–75 denoted a satisfactory level, and better than 75 was considered good.

Laboratory sample collection and transportation

A total of 242 samples containing four vegetable and two fruit types that are frequently consumed in the area were purchased weekly from randomly selected vendors in three selected markets. Variable numbers of samples: Lettuce (n = 42), Cabbage (n = 42), Green pepper (n = 42), Tomato (n = 40), Avocado (n = 38), and Mango (n = 38) were collected based on their abundance in markets. Nearly one sample of each vegetable and fruit type was collected per week per site for 12 weeks. Each sample was placed in a sterile polythene bag, properly labeled, and immediately transported to the Medical Microbiology Laboratory of Jimma University for bacteriological analysis. All samples were processed within 24 h after collection.

Microbiological analysis

Sample preparation and processing

We performed the microbiological analysis according to standard procedures. Briefly, 25 g of each vegetable sample and fruit sample was properly collected, trimmed, and transferred into a sterile sample bag containing 225 mL of Buffered Peptone Water (BPW) (CM1049) and incubated at 37 °C for 20 h to obtain a pre-enrichment homogenate. For selective enrichment of Salmonella spps., 0.1 mL and 1 mL of pre-enrichment culture were added to 10 mL of Rappaport Vassiliadis soya (RVS) broth and 10 mL of Muller Kauffmann tetrathionate-novobiocin (MKTTn) broth, and the plates were incubated at 41.50 °C and 37 °C for 24 h, respectively. Using a 10 µl microbiological loop, each selected enrichment homogenate sample was streaked on Xylose Lysine Deoxycholate (XLD) and Brilliance Salmonella Agar (OXOID, CM1092B + SR0194). The plates were allowed to solidify, inverted, and incubated at 37 °C for 24 h for colony formation. Salmonella spps. were isolated and identified based on distinctive morphological properties and using appropriate biochemical tests such as Triple Sugar Iron (TSI) agar test, urease test, and Sulphite-Indole-Motility (SIM) tests as recommended by EN ISO 6579-1:2017/Amd 1:2020 and as described by Hendriksen, R.S. (2003) [15, 16].

Escherichia coli O157: H7 detection

About 1 mL of pre-enrichment homogenate samples were added into a tube containing 9 mL of Modified Tryptone Soya Broth with Novobiocin (CM0989 + SR0181) and incubated for 18 h at 41.5 °C for selective enrichment [17, 18]. For selective isolation, 0.1 mL of selective enrichment samples were sub cultured on Cefixime-Tellurite Sorbitol MacConkey Agar (C-T SMAC) and on Cefixime-Rhamnose SMAC Agar (CR-SMAC) (CM1005 + SR0191) and incubated for 24 h at 37 °C. Non-sorbitol fermenter, transparent/colorless with a weak pale brownish E. coli O157:H7 suspect colonies were further confirmed with indole production tests and E. coli O157:H7 anti-sera latex agglutination test kits (DR0620M), following the manufacturer’s instructions. Escherichia coli O157 (CCUG 29889) and Escherichia coli (ATCC 25922) were used as positive and negative controls for reproducibility of Sorbitol MacConkey agar plates, respectively.

Antimicrobial susceptibility test

Antimicrobial susceptibility testing was conducted by the Kirby Bauer disk diffusion method according to the guidelines of clinical laboratory standards [19]. The media was prepared according to the instructions of the manufacturer. Briefly, bacterial suspensions were prepared in tubes containing 0.9% (w/v) phosphate-buffered saline with turbidity adjusted to 0.5 McFarland standard. Using a sterile cotton swab, bacterial suspension was streaked uniformly on the surface of Muller-Hinton agar. Antibiotic disks (from Oxoid (Basingstoke, England) including ciprofloxacillin (CIP-5 µg), Gentamycin (GM-10 µg), Ceftriaxone (CRO-30 µg), ceftazidime (CZT-30 µg), cefotaxime (CXT-30 µg), trimethoprim-sulfamethoxazole (SXT-1.25/23.75𝜇g), meropenem (MEM-10 µg), ampicillin (AMP-10 µg), amoxicillin-clavulanate (AMC-20/10µg) and chloramphenicol (CAF-30 µg) were placed on the surface of cultures. Finally, the diameter of the inhibition zone around the disks was measured after incubation of the plates at 37 °C for 24 h.

Phenotypic detections of ESBL production

The screening for ESBL producers was done by using a disc of ceftazidime (30 µg) and cefotaxime (30 µg). The zone of inhibition ≤ 22 mm for ceftazidime and ≤ 27 mm for cefotaxime was considered as potential ESBL producers as recommended by CLSI [19]. The combined disc test was used for phenotypic confirmation of ESBL producers. The combination of ceftazidime and cefotaxime alone and in combination with clavulanic acid (10 µg) were used for the confirmation of ESBL-producing isolates. An increase in the zone of inhibition ≥ 5 mm for either antimicrobial agent tested in combination with clavulanic acid versus its zone when tested alone was interpreted as positive for ESBL production. E. coli ATCC 25,922 and Salmonella Typhimurium ATCC 14,028 were used as reference strains for quality control. The strains were obtained from the Ethiopian Public Health Institute.

Data analysis and presentation

Data obtained from laboratory procedures were summarized and analyzed using a Microsoft Excel data sheet. Prevalence is expressed as the percent positive samples from the total samples tested. The numbers and proportions of positive samples and resistance isolates were presented using statistical tables.

Result

Socio-demographic characteristics and hygienic practices of vendors

During the visits, a total of 83 vegetable and fruit vendors were checked to see if they had a formal training certificate on food safety and on their vegetable and fruit handling practices by using observational checklists. About 71 (85.5%) of them were female, and 32 (38.6%) were 25–34 years old. Most vendors 23 (27.7%) didn’t complete their primary school, and none of them had any short course training on safe handling and processing of vegetables and fruits (Table 1).

In this study, only 30.0% of vegetable and fruit vendors had good knowledge on safety practices and sources of contamination. None of the vendors transported vegetables to the market or sold them under controlled temperature conditions. None of the vendors were washing and disinfecting their hands after handling money before handling vegetables again. About 36.1% of vendors placed their vegetables and fruits on dirty leaf materials, and 73.5% of them displayed openly without any protections during sale, which poses risk conditions for cross-contamination of fruits and vegetables. Washing of vegetables before display was practiced by 38.6%, among which 31.2% used stream water. In addition, 43.4% of them didn’t use separate packing materials for each type of fruit and vegetable. Moreover, most vegetable and fruit vendors indicated that inadequate supply of clean water and lack of clean, protected vendor sites posed a challenge towards maintaining hygiene (Table 2).

Table 1 Socio-demographic characteristics of vegetables and fruit vendors in peri urban kebeles of Jimma town, southwest Ethiopia, 2021
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Table 2 Hygiene and handling practices of vegetables and fruit vendors in peri urban kebeles of Jimma town, southwest Ethiopia, 2021
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Frequency of isolated bacteria

A total of 242 fresh vegetable and fruit samples were collected from local open markets and examined for bacterial contamination. Of these 242 samples tested, 12.8% (31/242) were contaminated with at least one pathogenic organism, Salmonella spp. and E. coli O157:H7. Among the vegetables taken from open markets, lettuce 21.4% (9/42) was the most frequently contaminated vegetable, followed by the green pepper 16.7% (7/42), and cabbage 14.3% (6/42). Ten-point five present (10.5%) of avocado and 5.3% (2/38) of mango were contaminated with the pathogenic organisms Salmonella spp. and E. coli O157:H7 (Table 3).

From the two pathogenic organisms recovered in this study, Salmonella spp. was detected in 10.7% (26/242) of the tested samples, whereas the overall detection rate of Escherichia coli O157:H7 was 2.1% (5/242). Salmonella spp. recovered in all types of samples, in which the detection rate was 21.4% (9/42) in lettuce samples, 16.7% (7/42) of green pepper, and 14.3% (6/242) of cabbage samples. Escherichia coli O157:H7 is detected in 2.8% (2/42) of sampled lettuce and in 2.4% (1/42) of each cabbage and green pepper samples, but not detected in sampled tomato and mango (Table 3).

Table 3 Distribution of Salmonella spps and E. Coli O157:H7 from fresh vegetable and fruit samples collected from peri urban kebeles of Jimma town, southwest Ethiopia
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Antimicrobial resistance profiles

In this study, 53.8% of Salmonella spps were resistant to ampicillin, 42.3% to co-trimoxazole, 46.2% to tetracycline, and 26.9% resistance was observed against ceftriaxone and cefotaxime. In addition, 23.1% and 19.2% of Salmonella isolates were resistant to ciprofloxacin and gentamicin, respectively. From the total of five E. coli O157:H7 isolates, 2 (40.0%) were resistant against ampicillin, amoxicillin-clavulanic acid, and cotrimoxazole. Only one isolate was resistant to ceftriaxone and cefotaxime, and no resistance was observed against ceftazidime, gentamicin, ciprofloxacin, chloramphenicol, and meropenem (Table 4).

Table 4 Antimicrobial resistance (AMR) profile of Salmonella spps and E. Coli O157:H7 isolated from fresh vegetable and fruit samples
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Multidrug resistance (MDR) profile of isolates

Out of the 26 Salmonella isolated, 6 (23.1%) that were isolated from salad showed multidrug resistance, and only one isolate of E. coli O157: H7 showed multidrug resistance (Table 5).

Table 5 Multidrug resistance (MDR) profile of isolates
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ESBL- producing isolates

A total of eight isolates (seven Salmonella and one E. coli O157:H7) fulfilled the screening criteria for ESBL production. Of these, four Salmonella and one E. coli O157:H7 isolate, with a total of 5/31 (16.1%) isolates, were confirmed as the ESBL producers (Table 6).

Table 6 Frequency of ESBL detection by combination disk-method
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Discussion

Vegetables and fruits are highly prone to contamination by microorganisms through contact with soil, water, and handling during harvest or after-harvest, and infections resulting from contamination of these products are serious health issues, particularly in underdeveloped nations like Ethiopia. In this study, the selected vegetable and fruit samples were taken from street vendors in three kebeles in Jimma town for the detection of pathogenic bacteria such as Salmonella spp. and E. coli 0157: H7 strain. Accordingly, a total of 12.8% of samples were contaminated with at least one of the pathogenic organisms, Salmonella spp. and/or E. coli O157:H7. Salmonella spps were detected in 10.7% (26/242) of the tested samples, whereas E. coli O157:H7 was detected in 2.1% (5/242) of the tested samples. The proportion of Salmonella positive samples in this study was lower than the previous study finding in different parts of Ethiopia: Mekelle (22%) [20], Fiche, Oromia (15%) [21], and Arba- Minch (13.3%) [22], and in Nepal (35.2%) [23], but higher than a finding in China (1% & 3%) [24, 25]. Similarly, the detection of E. coli O157:H7 in this study was 2.1%, which is slightly higher than a finding in Ethiopia, Addis Ababa (0.51%) [26], Nepal (1.4%) [23], and India (1.3%) [27]. The expected differences in the prevalence of these bacteria might be related to the difference in hygienic conditions of vendors, which could be risky for cross-contamination of fruit and vegetables from different contact surfaces during processing, transportation, and displaying at vendor sites. Poor handling, the sources, transportation, and storage temperature and period of the fruits and vegetables could contribute to the multiplication of bacteria [27]. In addition, different methods of detection, in sample size, the type of sample, and how and when it was collected could contribute to the observed variations.

The detection of Salmonella and E. coli O157:H7 in 25 gm of foods means that it is risky for consumption. Vegetables grown in soil fertilized by animal manure have a greater chance to be contaminated with E. coli O157: H7 [28, 29]. E. coli O157: H7 may survive in the soil from 7 to 25 weeks depending on soil types, humidity level, and temperature. Our study revealed that relatively higher proportions of lettuce samples were positive for Salmonella spp., followed by green pepper, cabbage, and avocado. In addition, although E. coli O157:H7 was also isolated from certain types of fresh vegetables and fruit with relatively low prevalence, this microorganism can cause illness in consumers. This may be potentially hazardous to consumers, especially for individuals with reduced immunity like patients with HIV/AIDS, pregnant women, young children, and old people.

It is well documented facts that a lack of education and training on food safety may contribute to unhygienic practices such as improper handling, processing, and display of fruit and vegetables at the vendor area [30, 31]. In this study, none of the vegetable and fruit vendors have a formal education certificate and/or short course training on food safety, which could be risky for cross-contamination of the fruits and vegetables with Salmonella spp. and E. coli O157:H7. This condition is serious in our situation because of the widespread practice of raw vegetable and fruit consumption throughout the country. Therefore, the range of activities should be carried out with the appropriate training on knowledge and hygienic practices of fruit and vegetable handlers.

Fruits and vegetables have been identified as vehicles for the transmission of pathogenic antimicrobial-resistant (AMR) microorganisms [32, 33]. In this study, the antimicrobial resistance pattern of isolates was checked with different classes of antibiotics. Accordingly, a total of 8 (6.6%) Salmonella spp. and E. coli O157:H7 isolates were resistant to two or more classes of antibiotics. More than half (53.8%) of Salmonella spps were resistant to ampicillin, 42.3% to co-trimoxazole, and 46.2% to tetracycline. Relatively lower rate, 26.9% of Salmonella spps were resistant to each of ceftriaxone and cefotaxime, 19.2% to gentamycin, and 23.1% to each of ciprofloxacin and chloramphenicol. The best efficacies of the above antibiotics were also reported in previous study findings in Ethiopia with different resistance rates [20, 34,35,36]. In contrast, the higher resistance rate of Salmonella spp. against tetracycline, ampicillin, cefotaxime, amoxicillin-clavulanic acid, ciprofloxacin, and gentamycin was reported in Ethiopia (21, 38), China [25], and India [38], in which the rate of resistance varies from 28.8 to 100%. The relatively high rate of resistance against ampicillin, tetracycline, and co-trimoxazole observed in this study might be related to inappropriate and excessive use of these antibiotics in humans and livestock. This study gives insight into the presence of antimicrobial-resistant pathogens in retailed and home-grown fruits and vegetables in our settings.

In the current study, 23.1% of the salmonella and 20.0% of the E. coli O157:H7 isolates were multi-drug resistant (MDR), which was also reported in the previous finding in Ethiopia (50.0%) [37], and Nepal (13.7%) [23]. The occurrence of drug and multidrug resistance isolates in these food sources means that they could be transmitted to consumers through the food chain, especially in our situation in which unhygienic practices are very prevalent. The frequency of ESBL producers for Salmonella was 15.4% and E. coli O157:H7 was 20.0% in the present study. It was also reported in a previous study in Nepal in which the prevalence of ESBL producers for Salmonella was 7.6% and 13.8% for E. coli isolates [23]. A similar study was conducted in Ethiopia and reported that 25.0% of ESBL producers Salmonella spp. were detected in vegetables [37]. The emergence of ESBL producers might also be related to inappropriate and excessive use of beta-lactam antibiotics.

Limitation of the study

The sample size for the study participants was not calculated. Instead, we used a time interval from first July to September 30, 2021, to get the required number of samples and those vendors from whom the required samples were collected during study period were included in the interview. The findings of this study may be reduced because we included some pathogenic bacteria from selected vegetables and fruits, making it impossible to conclude the true burden of the pathogenic bacteria in the study area. For more informative findings to indicate the specific antibiotic-resistant gene, a molecular technique was not conducted due to resource problems. However, the findings are still significant because the detection of these pathogenic bacteria in raw vegetables and fruits is concerning.

Conclusion

This study indicated that the safety aspect of fresh vegetable salads and fruit sold in Jimma town is unacceptable from a microbiological point of view. The presence of drug- and multi-drug-resistant Salmonella spp. and E. coli O157:H7 isolates was also a concerning problem. In addition, poor hygienic practices of vendors and unhygienic vending sites were observed, which may have great implications for cross-contaminations of vegetables and fruits. Therefore, provision of training and regular inspections to improve the knowledge and practice of vegetable and fruit vendors at roadside and retail shops about safe handling and distribution may have great implications in the prevention and control of the transmission of foodborne infections that might be caused by antibiotic-resistant strains.

Data availability

All the data supporting our findings were incorporated within the manuscript.

References

  1. World Health Organization. Promoting Fruit and Vegetable Consumption around the World. 2003. Available online: https://www.who.int/dietphysicalactivity/fruit/en/ (accessed on 28 November 2020).

  2. Berg G, Erlacher A, Smalla K, Krause R. Vegetable microbiomes: is there a connection among opportunistic infections, human health and our ‘gut feeling’? Microb Biotechnol. 2014;7(6):487–95. https://doiorg.publicaciones.saludcastillayleon.es/10.1111/1751-7915.12159

    Article  PubMed  PubMed Central  Google Scholar 

  3. FAO&WHO. Prevention and control of microbiological hazards in fresh fruits and vegetables – parts 1 & 2: General principles. Meeting Rep Microbiol Risk Assess Ser 42. 2023. https://doiorg.publicaciones.saludcastillayleon.es/10.4060/cc8490en. Rome.

    Article  Google Scholar 

  4. Hassan SA, Altalhi AD, Gherbawy YA, ElDeeb BA. Bacterial load of fresh vegetables and their resistance to the currently used antibiotics in Saudi Arabia. Foodborne Pathog Dis. 2011;8(9):1011–8.

    Article  PubMed  Google Scholar 

  5. Hölzel CS, Tetens JL, Schwaiger K. Unraveling the role of vegetables in spreading antimicrobial-resistant Bacteria: a need for quantitative risk Assessment. Foodborne Pathog Dis. 2018;15:671–88.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Estimating the burden. of foodborne diseases: a practical handbook for countries. Geneva.

  7. World Health Organization. 2021. Licence: CC BY-NC-SA 3.0 IGO.

  8. De Oliveira Elias S, Noronha TB, Tondo EC. Salmonella spp. and Escherichia coli O157:H7 prevalence and levels on letauce: A systematic review and meta-analysis. Food Microbiol. 2019;84:103217. https://doiorg.publicaciones.saludcastillayleon.es/10.1016/j.fm.2019.05.001

    Article  PubMed  Google Scholar 

  9. Gashaye D, Yildiz F. Wastewater-irrigated urban vegetable farming in Ethiopia: a review on their potential contamination and health effects. Cogent Food Agric. 2020;6(1). https://doiorg.publicaciones.saludcastillayleon.es/10.1080/23311932.2020.1772629

  10. Hölzel CS, Tetens JL, Schwaiger K. Unraveling the role of vegetables in spreading antimicrobial-resistant Bacteria: a need for quantitative risk Assessment. Foodborne Pathog Dis. 2018;15(11):671–88. https://doiorg.publicaciones.saludcastillayleon.es/10.1089/fpd.2018.2501

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Gazu L, Alonso S, Mutua F, Roesel K, Lindahl JF, Amenu K, Maximiano Sousa F, Ulrich P, Guadu T, Dione M, Ilboudo G, Grace D. Foodborne disease hazards and burden in Ethiopia: a systematic literature review, 1990–2019. Front Sustainable Food Syst. 2023;7:1058977. https://doiorg.publicaciones.saludcastillayleon.es/10.3389/fsufs.2023.1058977

    Article  Google Scholar 

  12. Asfaw T, Genetu D, Shenkute D, Shenkutie TT, Amare YE, Yitayew B. Foodborne pathogens and Antimicrobial Resistance in Ethiopia: an urgent call for action on one health. Infect Drug Resist. 2022;15:5265–74. https://doiorg.publicaciones.saludcastillayleon.es/10.2147/IDR.S375043

    Article  PubMed  PubMed Central  Google Scholar 

  13. Getaneh DK, Hordofa LO, Ayana DA, Tessema TS, Regassa LD. Prevalence of Escherichia coli O157:H7 and associated factors in under-five children in Eastern Ethiopia. PLoS ONE. 2021;16(1):e0246024. https://doiorg.publicaciones.saludcastillayleon.es/10.1371/journal.pone.0246024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Tadesse G. Prevalence of human salmonellosis in Ethiopia: a systematic review and meta-analysis. BMC Infect Dis. 2014;14:88. https://doiorg.publicaciones.saludcastillayleon.es/10.1186/1471-2334-14-88

    Article  PubMed  PubMed Central  Google Scholar 

  15. ISO 6579- Microbiology of the food. chain — Horizontal method for the detection, enumeration and serotyping of Salmonella — Part 1: Detection of Salmonella spp. — Amendment 1: Broader range of incubation temperatures, amendment to the status of Annex D, and correction of the composition of MSRV and SC (ISO 6579-1:2017/Amd 1:2020). https://standards.iteh.ai/catalog/standards/sist/22e687ba-56d5-4cda-bf00-4c87a6f5abdd/iso-6579-1-2017-amd-1-2020.

  16. Hendriksen RS. (2003) Laboratory Protocols Level 1: Training Course Isolation of Salmonella. A Global Salmonella Surveillance and Laboratory. Support Project of the World Health Organization, 4th Edition, WHO, Geneva.

  17. EN ISO 16654.:2001- Microbiology of food and animal feeding stuffs - horizontal method for the detection of Escherichia coli O157 - amendment 1: annex B: result of interlaboratory studies (ISO 16654:2001/Amd 1:2017). 2017-03.0-4.

  18. Hendriksen RS. (2003) Laboratory Protocols Level 4: Training Course Isolation and identification of enterohaemorrhagic Escherichia coli O157. A Global Salmonella Surveillance and Laboratory Support Project of the World Health Organization, 1st Edition, WHO, Geneva.

  19. CLSI. Performance standards for Antimicrobial susceptibility testing. 28th ed. PA: Clinical and Laboratory Standards Institute; 2018. CLSI supplement M100.

    Google Scholar 

  20. Berihu T, Gebremariam G, Weldu Y, et al. Prevalence, antimicrobial susceptibility test and associated factors of Salmonella and Shigella in ready-to-eat fruit juices and salads in Mekelle, northern Ethiopia. BMC Infect Dis. 2024;24:191. https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12879-024-09066-w

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Degaga B, Sebsibe I, Belete T, Asmamaw A. Microbial Quality and Safety of raw vegetables of Fiche Town, Oromia, Ethiopia. J Environ Public Health. 2022;2022. https://doiorg.publicaciones.saludcastillayleon.es/10.1155/2022/2556858

  22. Alemu G, Mama M, Siraj M. Bacterial contamination of vegetables sold in Arba Minch Town, Southern Ethiopia. BMC Res Notes. 2018;11:775. https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13104-018-3889-1

    Article  PubMed  PubMed Central  Google Scholar 

  23. Sapkota S, Adhikari S, Pandey A, et al. Multi-drug resistant extended-spectrum beta-lactamase producing E. Coli and Salmonella on raw vegetable salads served at hotels and restaurants in Bharatpur, Nepal. BMC Res Notes. 2019;12:516. https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13104-019-4557-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Cao C, Zhao W, Lü Z, Mo Y, Hu W, Sun S, Cheng H, et al. Microbiological analysis and characterization of Salmonella and ciprofloxacin-resistant Escherichia coli isolates recovered from retail fresh vegetables in Shaanxi Province, China. Int J Food Microbiol. 2023;387:110053. https://doiorg.publicaciones.saludcastillayleon.es/10.1016/j.ijfoodmicro.2022.110053

    Article  CAS  PubMed  Google Scholar 

  25. Qian M, Xu D, Wang J, Zaeim D, Han J, Qu D. Isolation, antimicrobial resistance and virulence characterization of Salmonella Spp. From fresh foods in retail markets in Hangzhou, China. PLoS ONE. 2023;18(10):e0292621. https://doiorg.publicaciones.saludcastillayleon.es/10.1371/journal.pone.0292621

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Haile AF, Alonso S, Berhe N, Bekele Atoma T, Boyaka PN, Grace D. Escherichia coli O157:H7 in Retail Lettuce (Lactuca sativa) in Addis Ababa City: Magnitude of Contamination and Antimicrobial susceptibility pattern. Front Microbiol. 2021;12:694506. https://doiorg.publicaciones.saludcastillayleon.es/10.3389/fmicb.2021.694506

    Article  PubMed  PubMed Central  Google Scholar 

  27. Mritunjay SK, Kumar V. Microbial quality, safety, and pathogen detection by using quantitative PCR of raw salad vegetables sold in Dhanbad city, India. J Food Protect. 2016;80(1):121–6.

    Article  Google Scholar 

  28. Liu D, Cui Y, Walcott R, Chen J. Fate of Salmonella enterica and Enterohemorrhagic Escherichia coli cells artificially internalized into Vegetable Seeds during Germination. Appl Environ Microbiol. 2018;84(1). https://doiorg.publicaciones.saludcastillayleon.es/10.1128/AEM.01888-17

  29. Islam M, Morgan J, Doyle MP, Phatak SC, Millner P, Jiang X. Fate of Salmonella enterica Serovar Typhimurium on carrots and radishes grown in fields treated with contaminated manure composts or irrigation water. Appl Environ Microbiol. 2004;70(4):2497–502.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Boakye MK, Tornyi JM, Dzubey I, Adzoyi P, Ayimah JC, Boakye DS, Wiafe ED. Assessment of food hygiene and safety knowledge, attitude, and practices of fruit and vegetable vendors in the Ho central market. Ghana Heliyon. 2023;9(9):e19579. https://doiorg.publicaciones.saludcastillayleon.es/10.1016/j.heliyon.2023.e19579

    Article  CAS  PubMed  Google Scholar 

  31. Ma L, Chen H, Yan H, et al. Food safety knowledge, attitudes, and behavior of street food vendors and consumers in Handan, a third-tier city in China. BMC Public Health. 2019;19:1128. https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12889-019-7475-9

    Article  PubMed  PubMed Central  Google Scholar 

  32. Olanbiwoninu A, Awotundun T, Olayiwola J, Somorin Y. Antimicrobial-resistant pathogens in fruits and vegetables from retail and home gardens. Sustainable Microbiol. 2024;1(1). https://doiorg.publicaciones.saludcastillayleon.es/10.1093/sumbio/qvad002

  33. Tenea GN, Reyes P, Molina D, Ortega C. Pathogenic microorganisms linked to Fresh fruits and juices purchased at low-cost markets in Ecuador, potential carriers of Antibiotic Resistance. Antibiotics. 2023;12(2):236. https://doiorg.publicaciones.saludcastillayleon.es/10.3390/antibiotics12020236

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Eromo T, Tassew H, Daka D, Kibru G. Bacteriological quality of street foods and antimicrobial resistance of isolates in Hawassa, Ethiopia. Ethiop J Health Sci. 2016;26(6):533–42.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Kibret M, Tadesse M. The bacteriological safety and antimicrobial susceptibility of bacteria isolated from street-vended white lupin (Lupinus albus) in Bahir Dar, Ethiopia. Ethiop J Health Sci. 2013;23(1):19–26.

    PubMed  PubMed Central  Google Scholar 

  36. Alelign D, Yihune M, Bekele M, Oumer Y, Beyene K, Atnafu K. Bacteriological quality and Antimicrobial resistant patterns of Foodborne pathogens isolated from commonly Vended Street foods in Arba Minch Town, Southern Ethiopia. Infect Drug Resist. 2023;16:2883–99. https://doiorg.publicaciones.saludcastillayleon.es/10.2147/IDR.S411162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Asfaw T, Genetu D, Shenkute D, Shenkutie TT, Yitayew B. Commonly consumed vegetables as a potential source of Multidrug-resistant and β-Lactamase-producing Bacteria in Debre Berhan Town, Ethiopia. Infect Drug Resist. 2023;16:3693–705. https://doiorg.publicaciones.saludcastillayleon.es/10.2147/IDR.S412126

    Article  PubMed  PubMed Central  Google Scholar 

  38. Sharma N, Singh K, Toor D, Pai SS, Chakraborty R, Khan KM. Antibiotic Resistance in microbes from Street Fruit drinks and Hygiene Behavior of the vendors in Delhi, India. Int J Environ Res Public Health. 2020;17:4829. https://doiorg.publicaciones.saludcastillayleon.es/10.3390/ijerph17134829

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We would like to thanks Jimma University for giving permission of the laboratory setup with necessary supplies. We also thank all the data collectors.

Funding

No funding was allocated for this study.

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Authors and Affiliations

  1. School of Medical Laboratory Sciences, Institute of Health, Jimma University, Jimma, Ethiopia

    Ahmed Zeynudin, Teshome Degefa, Tariku Belay, Abdulhakim Abamecha & Mengistu Abayneh

  2. Gumay Woreda Health Office, Jimma, Ethiopia

    Jiru Batu Mumicha

  3. Department of Oncology, Institute of Health, Jimma University, Jimma, Ethiopia

    Abdusemed Husen

  4. Oda Hulle Primary Hospital, Jimma, Ethiopia

    Jafer Yasin

  5. Division of Infectious Diseases and Tropical Medicine, University Hospital, Ludwig- Maximilians-Universität (LMU) Munich, Munich, Germany

    Andreas Wieser

  6. Department of Bacteriology, Max von Pettenkofer-Institute (LMU), Munich, Germany

    Andreas Wieser

  7. German Center for Infection Research (DZIF), Munich, Germany

    Andreas Wieser

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  1. Ahmed Zeynudin
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  9. Mengistu Abayneh
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Contributions

AZ, TD, TB, JBM, AH, JY, AA, AW and MA were participated in the study design, and were responsible for recruitment, sampling and for the laboratory analyses. AZ and MA analyzed the data and drafted the manuscript. All the authors have contributed to the manuscript and approved the final version.

Corresponding author

Correspondence to Mengistu Abayneh.

Ethics declarations

Ethical approval and consent to participate

Prior to the study implementation, ethical clearance was obtained from Institutional Ethics Board (IRB) of the Institute of Health, Jimma University, (Ref No: IHRPGn /357/2021). Participation was voluntary. Written informed consent was obtained from the sellers/vendors to participate in the study. The questionnaire was anonymous; therefore, any document as a written informed consent that might reveal the identity of the subjects was asked.

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Not applicable.

Competing interests

The authors declare that they have no competing interest.

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Zeynudin, A., Degefa, T., Belay, T. et al. Detections of antimicrobial resistance phenotypes and extended-spectrum beta-lactamase (ESBL)- producing Salmonella spps and Escherichia coli O157:H7 in raw vegetables and fruits from open markets in Jimma town, Ethiopia and evaluation of hygiene and handling practices of vendors. One Health Outlook 7, 2 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s42522-024-00125-5

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Keywords

One Health Outlook

ISSN: 2524-4655