Occurrence and antimicrobial susceptibility profiles of Escherichia coli and Salmonella spp. from unbranded and branded yogurt in Addis Ababa, Ethiopia

Abstract

The rapid growth of the Ethiopian dairy supply chain has raised concerns regarding food-borne pathogens and antibiotic-resistant bacteria. Therefore, it is crucial to have reliable methods for identifying bacteria and testing their susceptibility to antimicrobials in order to address public health concerns. The Vitek 2XL compact system is emerging as a promising tool due to its user-friendly nature and faster reporting times for microbial identification and antimicrobial susceptibility testing (AST). However, its application in yogurt analysis has been limited so far. To the best of our knowledge, this study is the first to use the Vitek 2XL system to assess the occurrence and antimicrobial susceptibility profiles of Escherichia coli, Salmonella spp., and Staphylococcus aureus isolates from unbranded and branded yogurt sold in Addis Ababa, Ethiopia. The findings from this research provide valuable insights into yogurt safety and aim to improve food safety practices in the region.
A total of 40 yogurt samples, consisting of 20 branded and 20 unbranded were collected from retail markets. The samples were cultured using International Organization for Standardization reference analytical methods. The VITEK 2 XL Compact system (bioMérieux) was employed to confirm the presumptive colonies and perform antimicrobial sensitivity testing, following the manufacturer’s instructions.
The overall prevalence of E. coli and Salmonella spp. was 14/40 (35%) and 3/40 (7.5%), respectively. However, S. aureus was not detected in the examined yogurt samples 0/40 (0%). The antimicrobial susceptibility testing revealed that 7% to 28% of E. coli isolates were resistant to 13 different antibiotics. However, high susceptibility was observed to Gentamicin (100%), Amikacin (93%), Marbofloxacin (93%), and Pardofloxacin (93%). For Salmonella spp., all isolates were resistant to Ampicillin (100%), Pardofloxacin (100%), Doxycycline (100%), and Tetracycline (100%). In contrast, 67% of Salmonella spp. isolates were susceptible to Amoxicillin/Clavulanic acid (67%), Cefpodoxime (67%), Cefovecin (67%), Ceftiofur (67%), and Neomycin (67%).
The results indicate that the tested yogurt samples may pose a risk of foodborne illnesses due to the presence of pathogenic bacteria. Additionally, the findings highlight a public health concern related to potential antibiotic resistance.

Full text

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Discover Food
Research
Occurrence andantimicrobial susceptibility profiles ofEscherichia coli
andSalmonella spp. fromunbranded andbranded yogurt inAddis
Ababa, Ethiopia
MihretFrew1,2· TakeleWolkaro2,4· UlnaGalmessa3
Received: 25 October 2024 / Accepted: 25 April 2025
© The Author(s) 2025 OPEN
Abstract
Background The rapid growth of the Ethiopian dairy supply chain has raised concerns regarding food-borne pathogens
and antibiotic-resistant bacteria. Therefore, it is crucial to have reliable methods for identifying bacteria and testing their
susceptibility to antimicrobials in order to address public health concerns. The Vitek 2XL compact system is emerging as
a promising tool due to its user-friendly nature and faster reporting times for microbial identication and antimicrobial
susceptibility testing (AST). However, its application in yogurt analysis has been limited so far. To the best of our knowl-
edge, this study is the rst to use the Vitek 2XL system to assess the occurrence and antimicrobial susceptibility proles
of Escherichia coli, Salmonella spp., and Staphylococcus aureus isolates from unbranded and branded yogurt sold in
Addis Ababa, Ethiopia. The ndings from this research provide valuable insights into yogurt safety and aim to improve
food safety practices in the region.
Methods A total of 40 yogurt samples, consisting of 20 branded and 20 unbranded were collected from retail markets.
The samples were cultured using International Organization for Standardization reference analytical methods. The VITEK
2 XL Compact system (bioMérieux) was employed to conrm the presumptive colonies and perform antimicrobial sen-
sitivity testing, following the manufacturer’s instructions.
Results The overall prevalence of E.coli and Salmonella spp. was 14/40 (35%) and 3/40 (7.5%), respectively. However,
S. aureus was not detected in the examined yogurt samples 0/40 (0%). The antimicrobial susceptibility testing revealed
that 7% to 28% of E.coli isolates were resistant to 13 dierent antibiotics. However, high susceptibility was observed
to Gentamicin (100%), Amikacin (93%), Marbooxacin (93%), and Pardooxacin (93%). For Salmonella spp., all isolates
were resistant to Ampicillin (100%), Pardooxacin (100%), Doxycycline (100%), and Tetracycline (100%). In contrast, 67%
of Salmonella spp. isolates were susceptible to Amoxicillin/Clavulanic acid (67%), Cefpodoxime (67%), Cefovecin (67%),
Ceftiofur (67%), and Neomycin (67%).
Conclusions The results indicate that the tested yogurt samples may pose a risk of foodborne illnesses due to the pres-
ence of pathogenic bacteria. Additionally, the ndings highlight a public health concern related to potential antibiotic
resistance.
Keywords Antibiotic· E. coli· Salmonella spp.· Vitek 2XL· Yoghurt
* Takele Wolkaro, takelewolkaro@yahoo.com | 1Ministry ofAgriculture, AddisAbaba, Ethiopia. 2School ofAnimal andRange Sciences,
Haramaya University, P. O. Box138, DireDawa, Ethiopia. 3Ethiopian Institute ofAgricultural Research, Holeta Agricultural Research,
Ethiopia, Holeta, Oromia, Ethiopia. 4College ofAgriculture andEnvironmental Sciences, School ofAnimal andRange Sciences, Haramaya
University, P.O. Box138, DireDawa, Ethiopia.

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1 Introduction
In Ethiopia, food safety governance is characterized by fragmented regulations, with overlapping responsibilities among
regulatory bodies and inconsistencies in enforcement and implementation [1–3], and there is a lack of eective surveil-
lance systems for monitoring foodborne pathogens to ensure food safety [2, 3]. Yoghurt, a widely consumed fermented
dairy product [4], is a valuable source of probiotics [5]. However, proper processing is crucial to produce safe and high-
quality yoghurt [6, 7]. Even small amounts of contamination can compromise yoghurt quality and adversely aect
consumer health [6, 7]. Previous studies in Ethiopia have shown that yoghurt can be a reservoir for various pathogenic
microorganisms that pose public health risks [8–12], and the fermentation process does not eectively control these
pathogens [13]. Therefore, yoghurt prepared from milk contaminated with these pathogens can pose a potential health
hazard, even in the absence of organoleptic changes [13]. Relying solely on sensory evaluation for assessing milk safety
is also insucient for ensuring its safety for consumption [13].
Studies have shown that the overuse and misuse of antimicrobial agents in food animal production have signicantly
contributed to the emergence of multidrug-resistant bacteria in Ethiopia [8–12, 14–16]. This has raised serious concerns
about the prevalence of major foodborne pathogens and antimicrobial resistance patterns in the rapidly expanding
Ethiopian dairy supply chain [3]. As a result, the research community has prioritized this concern, emphasizing the need
for rapid and reliable methods for bacterial identication and antimicrobial susceptibility testing of dairy products
[17]. In this context, the VITEK 2 XL Compact system has garnered attention for its ease of use and improved reporting
times for microbial identication and susceptibility testing in various applications, including wastewater treatment plant
euents [18]. However, its application in yoghurt sample analysis has been limited. To our knowledge, this study is the
rst to utilize the VITEK 2 XL Compact system to evaluate the occurrence and antimicrobial susceptibility proles of
Escherichia coli, Salmonella spp., and Staphylococcus aureus isolates from unbranded and branded yoghurt sold in Addis
Ababa, Ethiopia. The ndings from this research provide valuable insights into yogurt safety and aim to improve food
safety practices in the region.
2 Material andmethod
2.1 Description ofstudy area
The study was conducted in Addis Ababa, the capital city of Ethiopia, from February to May 2022. Addis Ababa is located
in the central highlands at an altitude of 2350m above sea level, at 9° 1′ 48″ N and 38° 44′ 24″ E. The city has an average
annual temperature of 16.3°C and receives an average annual precipitation of 1,143mm. Relative humidity ranges from
70 to 80% during the rainy season and 40% to 50% during the dry season, with bimodal rainfall averaging 1800mm [19].
2.1.1 Sample collection
The processing method for unbranded yoghurt has been previously described [20]. Unbranded yoghurt is a fermented
liquid milk product widely appreciated and consumed by all age groups across Ethiopia. Its production typically relies
on a natural fermentation process without controlled conditions such as milk standardization, culture concentration,
viability, incubation temperature, and time [20]. In contrast, branded yoghurt is produced under pasteurized conditions,
making it safer for consumption. However, most dairy processing plants in the country source raw milk from smallholder
producers, where basic food safety standards are often lacking [5]. A total of 20 branded yoghurt samples, representing
5 dierent brands with 4 samples per brand, were purchased from supermarkets using purposive sampling techniques.
Simultaneously, 20 unbranded yoghurt samples were collected randomly from 20 dierent dairy shops. The samples
were taken from each new batch twice a month over a 3-month period, from February to May 2022. Branded yoghurt
samples (250ml) were purchased before their expiration date, as indicated on the packaging, at the time of collection
and analysis. These samples were kept under refrigeration (2°C to 8°C) in their original silver or glass containers until
testing. Additionally, 250ml of unbranded yoghurt samples were collected aseptically in sterile screw-capped bottles
from dairy shops. Strict aseptic techniques, including the use of sterile materials, aming, and ice boxes, were applied
during all sampling and handling procedures. The yoghurt samples were labelled and transported to the Animal Prod-
ucts, Veterinary Drugs, and Feed Quality Testing Centre (APVDFQTC) in Addis Ababa in iceboxes containing ice packs,

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maintaining a temperature of 4°C, within 3 to 4h of collection. For ethical reasons, the samples were coded anonymously
throughout the study. Due to funding and logistical constraints, only 40 yoghurt samples were collected.
2.2 Isolation andidentification ofEscherichia coli
All samples were subjected to microbiological analysis to detect E. coli according to ISO 1665:2003 [21] with some modi-
cations and Quinn etal. [22]. Approximately 10g of yogurt was pre-enriched in 90ml of sterile buered peptone water
(BPW) (Oxoid Ltd., Basingstoke, UK) to yield a 1:10 dilution and incubated at 37°C for 24h. A loop full of the incubated
culture was streaked onto MacConkey agar (Oxoid Ltd., Basingstoke, UK) and incubated at 37°C for 24h. A single pink
colony from the MacConkey agar plate was sub-cultured on Eosin Methylene Blue (EMB) agar (Oxoid Ltd., Basingstoke,
UK) and incubated at 37°C for 24h. The presumptive E. coli colonies (green metallic sheen) were transferred onto nutrient
agar (Oxoid Ltd., Basingstoke, UK) and incubated at 37°C for 24h. A pure isolated single colony from a nutrient agar plate
was conrmed using the automated Vitek 2XL compact system (BioMérieux, Marcy-l’Étoile, France) using Gram Nega-
tive (GN) cards with 47 biochemical tests following the manufacturer’s instructions (BioMérieux, Marcy-l’Étoile, France).
2.3 Isolation andidentification ofStaphylococcus aureus
All samples were subjected to microbiological analysis to detect S. aureus according to ISO 6888-1:2021 [23] with some
modications. Approximately 10g of yogurt was pre-enriched in 90ml of sterile buered peptone water (BPW) (Oxoid
Ltd., Basingstoke, UK) to yield a 1:10 dilution and incubated at 37°C for 24h. A loop full of suspension was taken from
the incubated BPW culture, streaked on blood agar (Oxoid Ltd., Basingstoke, UK) and incubated at 37°C for 24h. A single
pink colony was taken from the incubated blood agar and sub-cultured on mannitol salt agar (MSA) (Oxoid Ltd., Basing-
stoke, UK) and incubated at 37°C for 24h. The presumptive S. aureus colonies (golden yellowish) were transferred onto
nutrient agar (Oxoid Ltd., Basingstoke, UK) and further incubated at 37°C for 24h. A pure isolated single colony from a
nutrient agar plate was conrmed using the automated Vitek 2XL compact system (BioMérieux, Marcy-l’Étoile, France)
with Gram-positive (GP) cards containing 43 biochemical tests, following the manufacturer’s instructions (BioMérieux,
Marcy-l’Étoile, France).
2.4 Isolation andidentification ofSalmonella species
All samples were subjected to microbiological analysis to detect Salmonella according to ISO 6579-1:2020 [24] with some
modications and Quinn etal. [22]. Approximately 25g of sample was pre-enriched in 225ml of buered peptone water
(BPW) (Oxoid Ltd., Basingstoke, UK) to yield a 1:10 dilution and incubated at 37°C for 24h. Then, the selective enrichment
media was prepared by mixing 1ml of the pre-enrichment culture with 10ml of Rappaport Vassiliadis (RVS) broth (Oxoid
Ltd., Basingstoke, UK) and incubated at 41.5°C for 24h. A loop full of culture from the selective enrichment broth was
transferred to selective agar media (xylose lysine deoxycholate agar (XLD) (Oxoid Ltd., Basingstoke, UK) and incubated
at 37°C for 24h. Presumptive Salmonella colonies (reddish or pink) were transferred onto nutrient agar (Oxoid Ltd.,
Basingstoke, UK) and further incubated at 37°C for 24h. A pure isolated single colony from a nutrient agar plate was
conrmed using the automated Vitek 2XL compact system (BioMérieux, Marcy-l’Étoile, France) with Gram Negative (GN)
cards containing 47 biochemical tests, following the manufacturer’s instructions (BioMérieux, Marcy-l’Étoile, France).
2.5 Antimicrobial susceptibility test (AST) withtheVitek 2XL compact system
The antimicrobial susceptibility test (AST) was conducted using the Vitek 2XL compact system, following the man-
ufacturer’s guidelines [25]. For this analysis, AST-GN96 (for E. coli) and AST-G79 (for Salmonella spp.) cards were
employed. These cards contained various antibiotics with specific minimum inhibitory concentration (MIC) ranges,
including Ampicillin (2–32µg/ml), Amoxicillin/Clavulanic acid (2–32µg/ml), Cephalexin (4–64µg/ml), Cefalotin
(4–64µg/ml), Cefpodoxime (0.25–8µg/ml), Cefovecin(0.12–8µg/ml), Ceftiofur (0.5–8µg/ml), Amikacin (4–64µg/
ml), Gentamicin (1–16µg/ml), Neomycin (8–128µg/ml), Enrofloxacin (0.5–8µg/ml), Marbofloxacin (0.5–4µg/mL),
Pardofloxacin (0.12–4µg/ml), Doxycycline (0.5–16µg/ml), Tetracycline (1–16µg/ml), Nitrofurantoin (16–256µg/ml),
Chloramphenicol (2–64µg/ml) and Trimethoprim/Sulfamethoxazole (20–320µg/ml). To prepare for testing, bacte-
rial suspensions of E. coli and Salmonella spp. were prepared in 3mL of sterile saline solution (0.45%–0.50% NaCl,
pH 6.5–7.0). The turbidity was adjusted using a DensiCHEK Plus meter to match the McFarland 0.5–0.63 standard.

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Each suspension was carefully mixed, and the corresponding AST-GN cards were assigned to E. coli and Salmonella
spp. by scanning them into the Vitek 2 FLEXprep data entry system. These cards were then loaded into the cassette
slot alongside the bacterial suspensions. The cassette was inserted into the Vitek 2XL system, which automatically
processed the samples. The system analyzed the susceptibility profiles based on its internal database and interpre-
tation algorithms, providing results within approximately 19h. The antimicrobial activity of each tested agent was
recorded in µg/mL, corresponding to the minimum inhibitory concentration (MIC) values according to Clinical and
Laboratory Standards Institute (CLSI) guidelines [26].
2.6 Statistical analysis
All collected data were recorded and managed using Microsoft Excel before being analyzed with SPSS version 21.
Descriptive statistics, including percentages and frequency distributions, were used to summarize bacterial isolate
occurrence and antimicrobial susceptibility. The susceptibility profiles were classified into resistant, intermediate,
or susceptible categories based on standard interpretations [26]. To assess the statistical significance of observed
differences, a chi-square (χ2) test was applied.
3 Results
3.1 The occurrence ofEscherichia coli, Salmonella spp. andStaphylococcus aureus
Among the 40 yogurt samples analyzed, Escherichia coli were detected in 14 samples (35%). When categorized by type,
9 out of 20 unbranded samples (45%) tested positive, compared to 5 out of 20 branded samples (25%). However, the dif-
ference between branded and unbranded yogurt samples was not statistically signicant (χ2 = 1.785, p = 0.185). Similarly,
Salmonella spp. was identied in 3 out of 40 yogurt samples (7.5%), with 2 positive cases (10%) in unbranded samples
and 1 case (5%) in branded samples. Again, the dierence was not statistically signicant (χ2 = 0.360, p = 0.548). In contrast,
Staphylococcus aureus was not detected in any of the 40 analyzed yogurt samples (0/40, 0%) (Table1).
3.2 Antimicrobial susceptibility pattern ofEscherichia coli andSalmonella spp.
Interestingly, the antimicrobial susceptibility test indicated that 7% to 28% of E. coli isolates were resistant to 13
antibiotics, while gentamicin, amikacin, marbofloxacine and pardofloxacin demonstrated susceptibility rates ranging
from 93 to 100%. Similarly, positive isolates of Salmonella spp. were 100% resistant to ampicillin, pardofloxacin, doxy-
cycline, and tetracycline, while 67% were susceptible to amoxicillin (clavulanic acid), cefpodoxime, cefovecin, ceftiofur
and neomycine (Table2).
Table 1 Occurrence of Escherichia coli, Salmonella spp. and Staphylococcus aureus from unbranded and branded yoghurt sold in Addis
Ababa, Ethiopia
Number outside the parenthesis indicate samples numbers while numbers in parenthesis are percentages
CI condence interval
Sample type Number tested E. coli Salmonella spp. S. aureus
Positive (95% CI) Negative Positive (95% CI) Negative Positive (95% CI) Negative
Unbranded yoghurt 20 9 (45%) (24.4–67.8) 11 (55%) 2 (10%) (1.2–31.7) 18 (90%) 0 (0–16.1) 20 (100%)
Branded yoghurt 20 5 (25%) (8.7–49.1) 15 (75%) 1 (5%) (0.1–24.9) 19 (95%) 0 (0–16.1) 20 (100%)
Total 40 14 (35%) (21.8–50.6) 26 (65%) 3 (7.5%) (1.6–20.4) 37 (92.5%) 0 (0–7.3) 100%
χ2 (p-value) 1.785 (0.185) 0.360 (0.548)

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4 Discussion
The detection of Escherichia coli in yoghurt samples in this study was comparable to findings by Aya etal. [27] in Ethio-
pia. In various studies, different prevalence rates of E. coli in yoghurt have been reported. For example, in Ethiopia,
Muluken etal. [28] found a prevalence of 0%, while Chaleshtori etal. [29] reported 10% in Iran. In Egypt, El-Ansary
[30] observed a prevalence of 12%, while in Ethiopia; Abebe etal. [8] recorded 13.9%. In another Ethiopian study,
Gugsa etal. [9] reported a prevalence of 22.9%. Similarly, in Egypt, Asfaw etal. [31] found a prevalence of 29.5%, while
Abike etal. [32] registered 23.5% in Ethiopia, and Bedasa etal. [10] reported 25.71% in Ethiopia. Conversely, studies
in other regions have recorded higher E. coli isolation rates from yoghurt. For instance, El-Malt etal. [33] in Nigeria
reported a prevalence of 44.8%, while Kandil etal. [34] observed a prevalence of 46.7% in Egypt. Awadallah etal.
[35] reported an exceptionally high prevalence of 88.0% in Egypt. The differences may be attributed to variations in
Table 2 Antimicrobial
susceptibility pattern of E. coli
and Salmonella spp. isolates
from unbranded and branded
yoghurt sold in Addis Ababa,
Ethiopia
Number outside the parenthesis indicate samples numbers while numbers in parenthesis are percentages
Antimicrobials Species Susceptible Intermediate Resistant
Ampicillin E. coli 11 (79%) – 3 (21%)
Salmonella spp. – – 3 (100%)
Amoxicillin/Clavulanic acid E. coli 12 (86%) 1 (7%) 1 (7%)
Salmonella spp. 2 (67%) – 1 (33%)
Cephalexin E. coli 12(86%) – 2 (14%)
Salmonella spp. 1 (33%) – 2 (67%)
Cefalotin E. coli 11 (79%) 1 (7%) 2 (14%)
Salmonella spp. 1 (33%) – 2 (67%)
Cefpodoxime E. coli 12 (86%) 1 (7%) 1 (7%)
Salmonella spp. 2 (67%) 1 (33%) –
Cefovecin E. coli 12 (86%) 2 (14%) –
Salmonella spp. 2 (67%) 1 (33%) –
Ceftiofur E. coli 12 (86%) 2 (14%) –
Salmonella spp. 1 (33%) 2 (67%) –
Amikacin E. coli 13 (93%) 1 (7%) –
Salmonella spp. 1 (33%) – 2 (67%)
Gentamicin E. coli 14 (100%) – –
Salmonella spp. 1 (33%) – 2 (67%)
Neomycine E. coli 12 (86%) 2 (14%) –
Salmonella spp. 2 (67%) 1 (33%) –
Enoroxacin E. coli 12 (86%) 1 (7%) 1 (7%)
Salmonella spp. – 1 (33%) 2 (67%)
Marbooxacine E. coli 13 (93%) – 1 (7%)
Salmonella spp. 1 (33%) 1 (33%) 1 (33%)
Pardooxacin E. coli 13 (93%) – 1 (7%)
Salmonella spp. – – 3 (100%)
Doxycycline E. coli 10 (72%) 2 (14%) 2 (14%)
Salmonella spp. – – 3 (100%)
Tetracycline E. coli 10 (72%) – 4 (28%)
Salmonella spp. – – 3 (100%)
Nitrofurantoin E. coli 12 (86%) – 2 (14%)
Salmonella spp. 1 (33%) – 2 (67%)
Chloramphenicol E. coli 10 (72%) 1 (7%) 3 (21%)
Salmonella spp. 1 (33%) – 2 (67%)
Trimethoprim/sulfamethoxazole E. coli 11 (79%) – 3 (21%)
Salmonella spp. 1 (33%) – 2 (67%)

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sampling procedures and detection methods employed. The increased prevalence of E. coli in yoghurt across vari-
ous studies may be attributed to factors such as the initial microbial load of the milk prior to processing, the use of
mastitic milk [36], inadequate sanitation of processing equipment, contamination from poor handling practices, or
the use of unhygienic packaging materials [37]. Additionally, storage conditions, including temperature and duration
from production to sampling, may also play a role in the microbial load [38]. Previous research has indicated that
E. coli infections from dairy products, including yoghurt and cheese made from raw milk, tend to be more severe,
resulting in higher hospitalization rates and more frequent instances of hemolytic uremic syndrome, with children
being more commonly affected compared to infections from beef consumption [22, 39].
The identication of isolated Salmonella spp. presented in Table1 showed that Salmonella spp. was detected in 3(7.5%)
out of 40 examined yoghurt samples. This result aligns with ndings from Mufandaedza etal. [40], who reported a preva-
lence of 15% in Zimbabwe. Conversely, higher prevalence rates were found by Abike etal. [32], with 44.8% prevalence
in Nigeria. In contrast, studies by Tesfaw etal. [41] and Lukásová etal. [42] reported the absence of Salmonella spp. in
yoghurt samples from Ethiopia and the Czech Republic, respectively. These dierences may be due to the inhibitory eect
that yoghurt cultures have on Salmonella spp., with inhibition rates ranging from 92.5% to 99.8% [43]. As per Ethiopian
regulations, yoghurt must be free of pathogenic microorganisms and adhere to microbiological limits, including the
absence of Salmonella [44]. The presence of Salmonella in food is generally recognized as a potential health hazard [14],
and thus, its occurrence in yoghurt poses a signicant public health risk [12].
Unbranded yoghurt is a fermented milk product widely consumed by all age groups throughout Ethiopia. Its produc-
tion generally involves a natural fermentation process, often without controlled conditions such as standardized milk,
culture concentration, viability, incubation temperature, and time [20]. The presence of E. coli and Salmonella spp. in
unbranded yoghurt samples suggests poor sanitary practices, which may result from the use of substandard raw milk
and the absence of a preheating step [45, 46]. In contrast, branded yoghurt is produced under pasteurized conditions,
ensuring a higher level of safety. However, many dairy processing plants in Ethiopia source raw milk from smallholder
producers, where fundamental food safety standards are often lacking [5]. The detection of E. coli and Salmonella spp.
in branded yoghurt samples suggests potential contamination after the processing stage [47, 48]. Thus, it is crucial to
prevent contamination during and after processing, and to ensure the quality of milk used in yoghurt production [49, 50].
S. aureus was not detected in examined yoghurt samples (Tables1). These ndings support reports from [51–53]. The
absence of S. aureus detection in this study may be due to factors such as the pH of yoghurt, its fat content, the pres-
ence of lactic acid, and inhibitory substances produced by yoghurt cultures, all of which limit or prevent the growth of S.
aureus in yoghurt [54]. In addition, these microorganisms were suppressed after a few days of storage of the fermented
product [55].
The antimicrobial susceptibility testing revealed a wide range of susceptibility and resistance patterns in the E. coli
isolates against the tested antimicrobial agents (Table2). Tetracycline resistance was the most frequently observed, with
4 isolates (28%) exhibiting resistance, which was higher compared to other antibiotics in this study. The resistance pat-
tern to tetracycline observed in the current study is similar to the ndings of Tadesse etal. (central Ethiopia) [56], who
reported a prevalence of 27.7%, but exceeds the results of Bekele etal. (Ethiopia) [57], who found a prevalence of 5.1%.
However, it is lower than the resistance levels reported by Gugsa etal. (Ethiopia) [9] with 41.7%, Messele etal. (Ethiopia)
[58] with 47.5%, Hailu (Ethiopia) [59] with 53.3%, Ababu etal. (Ethiopia) [60] with 60%, Ahmed and Shimamoto (Ethiopia)
[61] with 70%, and Disassa etal. (Ethiopia) [62] with 81.8%. In contrast, E. coli isolates exhibited high susceptibility to
amikacin, marbooxacin, pardooxacin, cefovecin, ceftiofur, neomycin, and enrooxacin with susceptibility rates ranging
from 86 to 93%. This nding is corroborated with reports from Kibret and Abera (central Ethiopia) [63] who found sus-
ceptibility rates in E. coli of 90%. Notably, all E. coli isolates were susceptible to gentamicin, which is commonly used, and
is consistent with the nding of Ababu etal. (Ethiopia) [60], who reported 100% susceptibility. This is higher than the
ndings of Kibret and Abera (Ethiopia) [63] who reported 81%, Gugsa etal. (Ethiopia) [9] who reported 91.7%, Bedasa
etal. (Ethiopia) [10] who reported 82.5%, and Hailu (Ethiopia) [59] who reported 81.82%.
Interestingly, all Salmonella spp. isolated in the current study were 100% resistant to ampicillin and tetracycline. This
nding is corroborated with reports by Fikirte etal. (Ethiopia) [12], who observed 100% resistance to these commonly
used antimicrobials, Addis etal. (Ethiopia) [15], who reported a prevalence of 95% resistance, and Geletu etal. (Ethiopia)
[63], who also found high resistance levels (88%) to ampicillin and tetracycline. The observed resistance of Salmonella spp.
to ampicillin and tetracycline may be attributing to the strains’ acquired ability to produce β-lactamase enzymes, which
can degrade the chemical structure of these antimicrobial agents [64]. Additionally, all Salmonella spp. isolates in the
current study showed high resistance to pardooxacin and doxycycline, which is greater than ndings reported by Bayeh
etal. (Ethiopia) [65] who observed 67% resistance to pardooxacin and Bayleyegn etal. (Ethiopia) [66] who reported 72%

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resistance to doxycycline. The eectiveness of gentamycin against Salmonella spp. isolates in this study was 33%, which
is lower than the 73.3%, 75%, and 100% reported by [15, 41, 67], respectively. However, a higher level of resistance to
gentamicin (75.6%) was reported in Gondar, Northwest Ethiopia by [68] which is contrary to the ndings in this study.
Studies conducted elsewhere in Ethiopia have indicated that the increase in antimicrobial-resistant bacterial strains
may be due to the irrational use of antimicrobials and inappropriate prescription and dispensing practices in both the
public veterinary and private health sectors of the country [12]. The scientic literature indicates that the presence of
multidrug-resistant bacterial pathogens in ruminants, and consequently in dairy products, could lead to drug-resistant
infections in humans [8–12]. To date, there are no published studies focusing on the occurrence and antimicrobial sus-
ceptibility proles of E. coli, Salmonella spp. and S. aureus isolates in yoghurt produced and commercialized in Ethiopia
with 2XL compact systems (bioMérieux) making broad comparisons of the results obtained dicult. The current study
addresses this gap.
5 Limitations ofthestudy
This study has limitations regarding the size, which may aect the accuracy of the estimates and reduce the strength
of the conclusions drawn in this study. Therefore, the present ndings encourage further investigations using a larger
sample size to enhance the reliability of the results.
6 Conclusion
The results from this study suggest that the yoghurt samples tested could pose a risk of foodborne illnesses due to the
presence of pathogenic bacteria and raise public health concerns related to antibiotic resistance. Therefore, it is important
to pay close attention to safety precaution and hygienic practice during yoghurt production, handling and marketing.
Additionally, the abuse and indiscriminate use of antibiotics in animals should be addressed.
Acknowledgements The authors would like to thank the Ministry of Ethiopia Agriculture and the Institute of Livestock Development for their
nancial support.
Author contributions "M.F. contributed to the writing of the research proposal and data collection. T.W. prepared the study design, data
analysis and interpretation, and manuscript writing. U.G. contributed to writing and editing the research proposal. All authors reviewed and
agreed on the nal draft of the manuscript. "
Funding This study was funded by Ethiopian meat and dairy Industry development Institute, Ethiopia.
Data availability All relevant data are within the paper. Raw data are available with the corresponding author upon request and also we are
trying to upload Science DB soon.
Declarations
Ethics approval and consent to participate There was no involvement of animals or humans for sample collection, as this study was conducted
on yoghurt samples obtained from ready-to-sell products in supermarkets and dairy shops. However, verbal consent was obtained from super-
markets and dairy shops owners after explaining the purpose and importance of the study prior to the start of data collection. Participation
of the supermarkets and dairy shops owners was entirely voluntary.
Consent for publication Not applicable.
Competing interests The authors declare no competing interests.
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which
permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to
the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modied the licensed material. You
do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party
material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If
material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds
the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:// creat iveco
mmons. org/ licen ses/ by- nc- nd/4. 0/.

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