Banner
Current IssueEditorial BoardOnline First Articles
Current IssueEditorial BoardOnline First Articles
Asian Journal of
Dairy and Food Research
Print ISSN: 0971-4456
Online ISSN: 0976-0563
NASS Rating
5.44
SJR
0.176
CiteScore
0.357

Chief Editor:
Harjinder Singh
Massey Institute of Food Science and Technology, NEW ZEALAND
Frequency:Bi-Monthly
Indexing:
Elsevier (Scopus and Embase), AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical...

Full Research Article

High Frequency of Antibiotic-resistant Enterobacterales Contaminating Chicken Viscera in Nigeria: A Trigger for Foodborne Disease Outbreak

Ikechukwu Benjamin Moses1, Emmanuel Nnabuike Ugbo1, Aswin Rafif Khairullah2, Wiwiek Tyasningsih3, Mustofa Helmi Effendi4,*, Chidimma Ruth Chukwunwejim5, Ukpai Ekenem Grace6
1Department of Applied Microbiology, Faculty of Science, Ebonyi State University, Abakaliki, Nigeria.
2Research Center for Veterinary Science, National Research and Innovation Agency (BRIN), Bogor, West Java, Indonesia.
3Division of Veterinary Microbiology, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, East Java, Indonesia.
4Division of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, East Java, Indonesia.
5Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Nigeria.
6Department of Microbiology and Parasitology, David Umahi Federal University of Health Sciences, Uburu, Ebonyi State, Nigeria.

ABSTRACT

Background: Most of the Enterobacterales strains are non-pathogenic; however, there is increasing concern that some antibiotic-resistant strains of Shigella spp., E. coli, Klebsiella spp. and Salmonella spp. could spread from animals and their by-products to humans and cause different diseases. This study aimed to isolate, characterize and assess the antimicrobial resistance profiles of clinically important Enterobacterales isolated from chicken viscera samples in Nigeria.

Methods: A total of 200 chicken viscera (intestine, gizzard, liver and crop) samples obtained from various poultry slaughterhouse outlets were analyzed using standard microbiological procedures. Using the disc diffusion technique, the detected Enterobacterales isolates were tested for antibiotic susceptibility.

Result: Results showed that Salmonella spp., E. coli, Klebsiella spp. and Shigella spp. were members of the Enterobacterales recovered from the chicken viscera samples with frequency of 88 (44%), 72 (36%), 60 (30%) and 32 (16%) respectively. Additionally, the highest frequency of Enterobacterales isolation was from chicken intestine with a frequency of 92/252 (36.5%). Generally, all the recovered Enterobacterales were resistant (100%) to tetracycline and erythromycin, except for Klebsiella spp in which 16.7% resistance frequency was observed. Resistances were also observed with ampicillin (37.5%-100%), gentamicin (34.4%-66.7%) and ciprofloxacin (12.5%-83.3%). The average multiple antibiotic resistance index (MARI) value of the recovered isolates in this study was 0.7. This study has shown that chicken viscera parts are contaminated by multidrug-resistant (MDR) Enterobacterales such as Shigella spp., E. coli, Klebsiella spp. and Salmonella spp.

INTRODUCTION

Chicken meats, including those from laying hens and broiler chickens, are among the livestock products that are frequently eaten. Interestingly, poultry viscera parts are also consumed by humans in many developing countries (Castellanos et al., 2018; Effendi et al., 2022; Hidayatik et al., 2024), including Nigeria. When two or more people contract the same disease after consuming comparable food tainted with bacteria, it’s known as a foodborne outbreak (Yanestria et al., 2019). It has been determined to be one of the main reasons why people get sick and die. Foodborne illnesses are now a threat to and a barrier to global economic development, thus every nation spends a great deal of effort and money fighting them (Robertson et al., 2018; Tyasningsih et al., 2022).
       
Enterobacterales, a group of Gram-negative, facultative anaerobic and rod-shaped bacteria are well-known common inhabitants of the gastrointestinal tracts of humans, poultry and other animals. The majority of Enterobacterales strains are not harmful, but there is growing worry that certain strains of Shigella spp., E. coli, Klebsiella spp. and Salmonella spp., which are resistant to antibiotics, may transfer from animals and their waste products to humans and cause a variety of illnesses like hemolytic-uremic syndrome, enteritis, endocarditis, meningitis and urinary tract infections, all of which can have fatal consequences. (Rahman et al., 2020; Ievy et al., 2022). Among chicken, the Enterobacterales group has been linked to coli granuloma, cellulitis, omphalitis, yolk sac infection, swollen head syndrome and colibacillosis (Islam et al., 2021; Gymoese et al., 2017).
       
During the handling, transportation and slaughter procedures, equipment such contaminated tables, cutting supplies and packaging can cause cross-contamination of chicken products (Fagbamila et al., 2017). Poor hygiene standards are the primary cause of the rising burden of foodborne illnesses, particularly in Africa. Microbial pathogens that cause foodborne illnesses may be acquired by food handlers through poor personal hygiene and inappropriate handling of meat products in abattoirs (Kusnoto et al., 2024). In poultry farming, antibiotics are frequently utilized as prophylactics, treatments, or growth promoters (Roy et al., 2022). The overuse and misuse of antibiotics is a major factor in the creation and spread of antibiotic-resistant Enterobacterales, which can infect people through food or direct contact with ill animals (Founou et al., 2016; Wibisono et al., 2021).
       
The evolution of antimicrobial resistance (AMR) is the most controversial topic affecting the health of people, animals and ecosystems in the twenty-first century (Urmi et al., 2021; Yanestria et al., 2022). Antimicrobial resistance constitutes one of the greatest challenges in modern medicine as it compromises and limits therapeutic options for the treatment of bacterial infections (Riwu et al., 2022; Hemamalini et al., 2025). Various mechanisms can lead to antimicrobial resistance in Enterobacterales, such as drug-expelling active efflux pumps, decreased cellular permeability, increased production of enzymes that modify or inactivate medicines and target mutations that result in modification (Ranjbar and Farahani, 2019). Investigating and understanding the contamination of poultry products by antibiotic-resistant Enterobacterales contributes to enhancing food safety as this helps in implementing measures to minimize the risk of foodborne illnesses associated with the consumption of contaminated poultry (Putri et al., 2023).
       
Although, there are some reports on bacterial contamination of food-producing animals and their by-products; it is imperative to keep track of the current spread of antibiotic-resistant bacterial pathogens in order to develop strong preventing measures that will curtail AMR spread in both veterinary and human medicine. Additionally, there is still paucity of publicly available information on the public health implications of using poultry viscera as human food without proper processing, especially in developing countries, including Nigeria. This study, was therefore designed to isolate, characterize and determine the frequency of antibiotic-resistant Enterobacterales contaminating chicken viscera sold in different poultry slaughterhouse outlets in Abakaliki, Southeastern Nigeria.

MATERIALS AND METHODS

Collection of samples
 
A total of 200 chicken viscera samples of intestine (50), gizzard (50), liver (50) and crop (50) were aseptically collected from broiler chickens in various poultry slaughterhouse outlets between July and November, 2023. All aseptically collected chicken viscera parts were firstly cut open with small sterile scalpel blade and thereafter, swabs of the incised viscera parts were taken using sterile cotton swab sticks. Collected swab samples were labelled and placed in sample bags and then transported on ice packs within one hour of collection to the Department of Applied Microbiology laboratory, Ebonyi State University, Abakaliki, Nigeria for bacteriological analysis.
 
Processing of samples and bacteriological analysis
 
Prior to being incubated for 18 to 24 hours at 37°C, collected swabs were first enriched in buffered peptone water (Oxoid, Hampshire, UK). Following the transfer of a loop full of the inoculated peptone water onto Salmonella-Shigella (SS) agar (used to isolate Salmonella spp. and Shigella spp.), MacConkey agar (used to isolate Klebsiella spp.) and Eosine Methylene Blue (EMB) agar (used to isolate E. coli) plates, the plates were incubated for 24 hours at 37°C. All culture media were purchased from Oxoid, Hampshire, UK. Colonies typical of Salmonella spp. (colourless colonies with dark centre on SS agar), Shigella spp. (smooth colourless colonies on SS agar), E. coli (greenish-metallic sheen on EMB agar) and Klebsiella spp. (mucoid pink-coloured colonies on MacConkey agar) were respectively sub-cultured through successive streaking in order to obtain pure colonies which were then subjected to further physiological and biochemical characterization such as Gram-staining, catalase, indole, oxidase, methyl red, citrate utilization, urease, Voges-Proskauer and nitrate reduction tests (Cheesbrough, 2006; Moses et al., 2018; Pradika et al., 2019; Widodo et al., 2022). Pure colonies of identified isolates were then preserved and stored on nutrient agar slant at 4°C for further tests.
 
Antimicrobial susceptibility testing
 
The Clinical and Laboratory Standards Institute’s (CLSI, 2022) criteria for the disc diffusion method on Mueller-Hinton agar were followed in order to determine the antimicrobial susceptibility tests. Mueller-Hinton (MH) agar (Oxoid, UK) was produced in compliance with the guidelines provided by the manufacturer. The medium was cooled, poured into plates and allowed to solidify before use. The test isolate’s broth culture, which was 18-24 hours old, was standardized to the 0.5 McFarland standard. The standardized inoculums were placed inside a sterile swab stick, which was then drained to remove any extra inoculum load. The inoculums were then spread out onto a Mueller-Hinton agar plate surface. The Mueller-Hinton agar plate that had been inoculated was closed and left to dry at room temperature for a short while. The following antimicrobials (Oxoid, UK) were tested against the test isolates: ciprofloxacin (5 µg), erythromycin (15 µg), gentamicin (10 µg), ampicillin (10 µg) and tetracycline (30 μg). Using sterile forceps, all antibiotic discs (Oxoid, UK) were meticulously placed on the Mueller-Hinton agar plates that had been infected. Following the inoculation, the MH agar plates were incubated for 18 to 24 hours at 37°C in an aerobic environment. After incubation, inhibition zone diameters were measured, recorded and categorized as resistant, intermediate, or susceptible using the CLSI standards (CLSI, 2022). In this investigation, isolates with intermediate resistance were categorized as “resistant”.
 
Determination of multiple antibiotic resistance index (MARI)
 
Each isolate’s MARI value was determined using the disk diffusion method’s interpreted findings. MARI was computed by dividing the total number of antibiotics tested against the isolate (y) by the number of antibiotics to which the isolate shows resistance (x). It is expressed as MARI = x/y (Moses et al., 2020; Moses et al., 2022).
 
Statistical analysis
 
The statistical evaluation of categorical variables was done with the SPSS version 17.0 statistical software package (SPSS Inc., Chicago, USA) using one-way between subjects ANOVA tool. The p-value of less than 0.05 (pvalue < 0.05) was deemed statistically significant for the results.

RESULTS AND DISCUSSION

Results showed that Salmonella spp. (Fig 1), E. coli (Fig  2), Klebsiella spp. (Fig 3) and Shigella spp. (Fig 4) were members of the Enterobacterales recovered from the 200 chicken viscera samples with frequencies of 88 (44%), 72 (36%), 60 (30%) and 32 (16%) respectively (Table 1). Additionally, the highest frequency of Enterobacterales isolation was from chicken intestines with a frequency of 92/252 (36.5%). This was closely followed by the crop 77/252 (30.6%), liver 43/252 (17.1%) and gizzard 40/252 (15.9%) (Table 1).

Fig 1: Salmonella spp. (colourless colonies with dark centre) on SS agar.



​

Fig 2: E. coli (greenish-metallic sheen colonies) on EMB agar.



​

Fig 3: Klebsiella spp. (mucoid pink-coloured colonies) on MacConkey agar.



​

Fig 4: Shigella spp. (smooth opaque colourless colonies) on SS agar.



​

Table 1: Distribution of Enterobacterales isolated from different visceral parts of chicken.


       
A one-way between subjects ANOVA was conducted to determine if the isolation frequency of Enterobacterales recovery is dependent on the type and nature of chicken viscera parts. Statistical analysis results showed that there was no statistically significant difference in the frequency of recovered Enterobacterales between the crop, gizzard, intestine and liver parts of chicken viscera that were analyzed in this study [F (3, 12) = 1.142, p = 0.372].
       
A one-way between subjects ANOVA was also conducted to determine if there is a statistically significant difference in the frequency of each major strains of Enterobacterales recovered from chicken viscera. Results also showed that there was no statistically significant difference in the frequency of recovered E. coli, Klebsiella spp., Shigella spp. and Salmonella spp from the viscera parts of chicken [F (3, 12) = 0.934, p = 0.454].
       
Generally, all the recovered Enterobacterales (Salmonella spp., E. coli, Klebsiella spp. and Shigella spp.) were resistant (100%) to tetracycline and erythromycin, except for Klebsiella spp in which 16.7% resistance frequency was observed (Table 2). Resistances were also observed with ampicillin [(Shigella spp, 100%; E. coli, 37.5% and Salmonella spp., 37.5%)], gentamicin [(Klebsiella spp, 66.7%; Shigella spp., 34.4%)] and ciprofloxacin [(Klebsiella spp, 83.3%; Shigella spp, 34.4%; E. coli, 12.5% and Salmonella spp., 12.5%)]. The multiple antibiotic resistance index (MARI) value of the recovered isolates in this study ranged from 0.64-0.75 (Table 2).

​

Table 2: Antimicrobial susceptibility profiles of Enterobacterales isolated from different visceral parts of chickens.


       
A one-way between subjects ANOVA was also conducted to determine if there is a statistically significant difference in the resistance frequency of each major strains of Enterobacterales recovered from chicken viscera. From the results, no statistically significant difference was observed when the mean resistance frequencies of the recovered E. coli, Klebsiella spp., Shigella spp. and Salmonella spp. were compared [F (3, 16) = 0.340, p = 0.797].
       
Members of the Enterobacterales group such as E. coli, Klebsiella spp., Salmonella spp. and Shigella spp. are ubiquitous in nature and have been recognized as common inhabitants of the intestinal tracts of food-producing animals. Importantly, they are well-known foodborne pathogens associated with foodborne illnesses in humans and also the cause of mortality in commercial poultry and other food-producing animals (Ewers et al., 2004; Pradika et al., 2019; Widodo et al., 2022).
       
In this study, different strains of the Enterobacterales group, especially Salmonella spp., E. coli, Klebsiella spp. and Shigella spp. were recovered from 200 chicken viscera samples with isolation frequency of 88 (44%), 72 (36%), 60 (30%) and 32 (16%) respectively. Interestingly, the frequency of Enterobacterales isolation was highest in chicken intestines with a frequency of 92/252 (36.5%). This was closely followed by the crop 77/252 (30.6%), liver 43/252 (17.1%) and gizzard 40/252 (15.9%). Additionally, all the recovered Enterobacterales (Salmonella spp., E. coli, Klebsiella spp. and Shigella spp.) were generally resistant (100%) to tetracycline and erythromycin, except for Klebsiella spp in which 16.7% resistance frequency was observed. Resistances (12.5%-100%) were also observed to ciprofloxacin, gentamicin and ampicillin. Isolates were multidrug-resistant with a mean multiple antibiotic resistance index (MARI) value of 0.7.
       
The frequency of E. coli (20.8%) recovered from liver in our study is higher than the 13.6% reported by Sarba et al., (2019). Interestingly, just like we reported E. coli prevalence in other chicken visceral parts such as gizzard (13.9%), crop (20.8%) and intestine (44.4%), Sarba et al., (2019) also reported E. coli in different chicken internal organs/parts such as the kidney (6.3%), spleen (15.2%) and ovary (10.7%). Dashe et al., (2003) also found that the isolation rate of E. coli was 15.8% from the liver and 13% from the spleen, indicating that E. coli is primarily found in these organs.
       
The frequency of Klebsiella spp, (30%) in our study also agrees with the study of Wareth and Neubauer (2021) who reported the isolation of Klebsiella spp. from poultry carcass and internal organs; although with a higher isolation frequency of 45%.
       
Shigella spp. was the least prevalent Enterobacterales recovered from chicken viscera samples analysed in our study with a frequency of 16%. Other studies have also reported Shigella spp. in animal products with frequency ranging from 1.4%-7% (Elkenany et al., 2022; Ahmed and Shimamoto, 2014). Our study is also in concord with another study that reported Salmonella spp. in poultry products (Gad et al., 2018).
       
There was no statistically significant difference in the frequency of recovered Enterobacterales between the crop, gizzard, intestine and liver viscera parts of chicken that were analyzed in this study (p=0.372). Although more Enterobacterales pathogens were recovered from the chicken intestine and crop; the differences in the frequency of Enterobacterales isolation in the 4 chicken viscera parts that were analysed is likely due to chance and not because of the nature of the chicken viscera parts.  Additionally, there was no statistically significant difference in the frequency of recovered E. coli, Klebsiella spp., Shigella spp. and Salmonella spp from the viscera parts of chicken (p = 0.454). The recovery frequency of the Enterobacterales strains in this study is also likely due to chance.
       
The differences in isolation frequency of Enterobacterales in our study when compared with other studies might likely be due to differences in sample size, hygienic practices of the poultry product vendors and farmers, the sampling time/period and location. Possible sources of additional contamination of the chicken viscera might be the chicken itself, or from contamination during the processing (such as dressing, packaging, storage etc.), as good aseptic conditions were ensured from sample collection point to laboratory analysis during this study.
       
Antibiotic-resistant strains of the Enterobacterales group have also been reported to be common bacterial contaminants in retail poultry meat and other animal products (Anderson et al., 2019) with significant public health threat and economic burden. Antibiotic-resistant bacterial pathogens such as E. coli and methicillin-resistant Staphylococcus aureus (MRSA) have also been implicated in the contamination of other animal products, especially cow milk in Indonesia (Pradika et al., 2019; Widodo et al., 2022; Khairullah et al., 2022). A minimum of three distinct types of antibiotics were resistant to the isolated Enterobacterales in this investigation, indicating their multidrug resistance. Importantly, the recovered Enterobacterales (Salmonella spp., E. coli, Klebsiella spp. and Shigella spp.) exhibited resistance (12.5%-100%) to ciprofloxacin, gentamicin, ampicillin, erythromycin and tetracycline. Additionally, the average multiple antibiotics resistance index (MARI) value of the Enterobactaerales strains in this study ranged from 0.64-0.75 with a mean MARI value of 0.7; thus, depicting the misuse of antibiotics in our study area. The antimicrobial susceptibility testing results in this study showed varying patterns of responses to tested antibiotics by the different strains of the Enterobacterales group. Interestingly, some antimicrobials (especially ampicillin and gentamicin) that were completely active (100%) against all the recovered E. coli and Salmonella spp. isolates were ineffective against some other Enterobacterales strains of Klebsiella spp. and Shigella spp. which exhibited resistance frequencies of 66.7% and 34.4% respectively.
       
Similar patterns of antimicrobial resistance, including multidrug-resistant (MDR) strains that were observed in our study have also been reported in other studies. Sarba et al., (2019) reported high frequency of MDR E. coli recovered from chicken internal organs. In their study, resistances were noted for fluoroquinolones (100%), aminoglycosides (69.9-89%), beta-lactams (84.6%) and macrolides (75%). Our study also agrees with the work of Adenipekun et al., (2015) who also isolated MDR E. coli in chicken products. Anderson, (2019) reported that all the Shigella spp. isolates recovered from chicken products were resistant to beta-lactam antibiotics. Okoli et al., (2021) also reported that all the Salmonella and Shigella isolates recovered from chickens in their study were resistant to the β-lactam antibiotics. MDR Enterobacterales recovered from chicken products with similar antibiotic resistance patterns to the isolates in our study have also been reported in by other authors (Gu et al., 2015; Teimourpour et al., 2019; Ben et al., 2019; Elkenany et al., 2022; Shoja et al., 2023).
       
No statistically significant difference was observed when the mean resistance frequencies of the recovered Salmonella spp, Klebsiella spp., E. coli and Shigella spp. in this study were compared (p=0.797). Their different resistance frequency is likely due to chance. Results of this study further emphasizes the importance of conducting antimicrobial susceptibility testing, especially in cases of foodborne illnesses/diseases, in order to determine the most effective therapeutic antimicrobial options and also to curtail the increasing spread of antimicrobial resistance in both human and veterinary medicine which might result from wrong/indiscriminate antimicrobial prescription/usage (Yanestria et al., 2024).
       
The widespread occurrence of antimicrobial resistance within or between antibiotic classes in different studies may be the result of incorrect or careless antimicrobial usage in human, veterinary and agricultural medicine (Khairullah et al., 2024). Similar antibiotics are often used indiscriminately over extended periods of time, particularly in poultry farms, which may encourage the formation and spread of bacteria resistant to antibiotics (Riwu et al., 2024). This is why, today, antimicrobial resistance has become a serious growing global public health problem in both veterinary and human medicine (Li et al., 2021).
       
To reduce the risk of contamination by MDR Enterobaterales, improved hygiene and sanitation practices during poultry production and processing should be implemented. This may include improved sanitation of poultry housing and slaughtering facilities, as well as better hygiene practices by poultry workers. These measures could help reduce the risk of foodborne disease outbreak.

CONCLUSION

This study has shown that chicken viscera parts sold in poultry slaughterhouse outlets in Abakaliki, Nigeria are highly contaminated with multidrug-resistant (MDR) Enterobaterales. Among the recovered Enterobacterales, Salmonella spp. was the most prevalent (44%), followed by E. coli (36%), Klebsiella spp. (30%) and Shigella spp. (16%). Interestingly, the highest frequency of Enterobacterales isolation was from chicken intestines 92/252 (36.5%). Generally, all the recovered Enterobacterales were resistant (100%) to tetracycline and erythromycin, except for Klebsiella spp in which 16.7% resistance frequency was observed. Resistance was also observed with ampicillin (37.5%-100%), gentamicin (34.4%-66.7%) and ciprofloxacin (12.5%-83.3%). The detection of MDR Enterobacterales in chicken viscera as observed in our study calls for strong measures that would safeguard human health and also curb the increasing incidence and spread of superbugs in the food chain which could be accompanied by serious public health consequences, especially foodborne disease outbreaks. Additionally, public health advocacy such as sensitization of poultry farmers on good hygienic practices, prudent use of antibiotics and the continuous monitoring of poultry farms and other outlets where poultry products are sold to the public for MDR strains of Enterobacterales are very vital in the containment of foodborne diseases.

ACKNOWLEDGEMENT

The authors would like to acknowledge Ebonyi State University, Abakaliki Nigeria and Lembaga Penelitian dan Pengabdian Masyarakat Universitas Airlangga, Indonesia for their support. This study was partly supported by the International Research Consortium, Lembaga Penelitian dan Pengabdian Masyarakat, Universitas Airlangga, Surabaya, Indonesia Year 2024 with grant number: 171/UN3.LPPM/PT.01.03/2024.

CONFLICT OF INTEREST

All authors declare that they have no conflict of interest.

REFERENCES


  1. Adenipekun, E.O, Jackson, C.R., Oluwadun, A., Iwalokun, B.A., Frye, J.G., Barrett, J.B., Hiott, L.M., Woodley, T.A. (2015). Prevalence and antimicrobial resistance in Escherichia coli from food animals in Lagos, Nigeria. Microbial Drug Resistance. 21(3): 365-372. doi: 10.1089/mdr.2014.0222.

  2. Ahmed, A.M., Shimamoto, T. (2014). Isolation and molecular chara- cterization of Salmonella enterica, Escherichia coli O157:H7 and Shigella species from meat and dairy products in Egypt. International Journal of Food Microbiology. 168-169(1): 57-62. doi: 10.1016/j.ijfoodmicro.2013.10.014.

  3. Anderson, M., Smith, J., Johnson, L. (2019). High prevalence of multidrug-resistant Klebsiella species in clinical isolates. A major healthcare challenge. The Journal of Infectious Diseases. 67(3): 215-222. doi: 10.3390/antibiotics100 81007.

  4. Ben, Y., Fu, C., Hu, M., Liu, L., Wong, M.H., Zheng, C. (2019). Human health risk assessment of antibiotic resistance associated with antibiotic residues in the environment: A review. Environmental Research. 169(1): 483-493. doi: 10.1016/ j.envres.2018.11.040.

  5. Castellanos, L.R., van der Graaf-van Bloois, L., Donado-Godoy, P., León, M., Clavijo, V., Arévalo, A., Bernal, J.F., Mevius, D.J., Wagenaar, J.A., Zomer, A., Hordijk, J. (2018). Genomic characterization of extended-spectrum cephalosporin- resistant salmonella enterica in the colombian poultry Chain. Frontiers in Microbiology. 9(1): 11-97. doi: 10. 3389/fmicb.2018.02431.

  6. Cheesbrough, M. (2006). District Laboratory Practice in Tropical Countries (Part II), Cambridge University. 19-110. doi: 10. 1017/CBO9780511543470.

  7. Clinical Laboratory Standards Institute (CLSI). (2022). Performance Standards for Antimicrobial Susceptibility Testing. 32nd Edition, CLSI supplements M100Wayne. PA, USA, 2022; Available online: https://clsi.org/

  8. Dashe, Y., Raji, M., Abdu, P., Oladele, B. (2003). Distribution of aerobic bacteria in visceral organs of sick and apparently healthy chickens in Jos, Nigeria. International Research Journal of Microbiology. 4(3): 79-83. 

  9. Effendi, M.H., Faridah, H.D., Wibisono, F.M., Wibisono, F.J., Fatimah, Nisa, N., Ugbo, E.N. (2022). Detection of virulence factor encoding genes on Escherichia coli isolated from broiler chicken in Blitar District, Indonesia. Biodiversitas. 23(7): 3429-3436. doi: 10.13057/biodiv/d230716.

  10. Elkenany, R., Eltaysh, R., Elsayed, M., Abdel-Daim, M., Shata, R. (2022). Characterization of multi-resistant Shigella species isolated from raw cow milk and milk products. Journal of Veterinary Medical Science. 84(7): 890-897. doi: 10. 1292/jvms.22-0018.

  11. Ewers, C., Janben, T., Kiebling, S., Philipp, H.C., Wieler, L.H. (2004). Molecular epidemiology of avian pathogenic Escherichia coli (APEC) isolated from colisepticemia in poultry. Veterinary Microbiology. 104(1-2): 91-101.

  12. Fagbamila, I.O., Barco, L., Mancin, M., Kwaga, J., Ngulukun, S.S., Zavagnin, P., Lettini, A.A., Lorenzetto, M., Abdu, P.A., Kabir, J., Umoh, J., Ricci, A., Muhammad, M. (2017). Salmonella serovars and their distribution in Nigerian commercial chicken layer farms. PLoS One. 12(3): e0173097. doi: 10.1371/journal.pone.0173097.

  13. Founou, L.L., Founou, R.C., Essack, S.Y. (2016). Antibiotic resistance in the food chain: A developing country-perspective. Frontiers in Microbiology. 7(1): 1881.

  14. Gad, A.H., Abo-Shama, U.H., Harclerode, K.K., Fakhr, M.K. (2018). Prevalence, serotyping, molecular typing and antimicrobial resistance of Salmonella isolated from conventional and organic retail ground poultry. Frontiers in Microbiology. 91(1): 261-553.  

  15. Gu, B., Zhou, M., Ke, X. (2015). Comparison of resistance to third- generation cephalosporin’s in Shigella between Europe- America and Asia-Africa from 1998 to 2012. Epidemiology and Infection. 143(13): 2687-2699. doi: 10.1017/S0950 268814003446.

  16. Gymoese, P., Sørensen, G., Litrup, E., Olsen, J.E., Nielsen, E.M., Torpdahl, M. (2017). Investigation of outbreaks of Salmonella enterica Serovar typhimurium and its monophasic variants using whole-genome sequencing, Denmark. Emerging Infectious Diseases. 23(10): 1631-1639. doi: 10.3201/ eid2310.161248.

  17. Hemamalini, N., Shanmugam, S.A., Kathirvelpandian, A., Deepak, A., Kaliyamurthi, V., Suresh, E., Ezhilmathi, S. (2025). Prevalence, antimicrobial resistance and resistance gene cassettes detection in bacterial pathogens isolated from freshwater ornamental fishes. Indian Journal of Animal Research. 59(3): 467-475. doi: 10.18805/IJAR.B-4903.

  18. Hidayatik, N., Yuliani, G.A., Agustono, B., Kristianingtyas, L., Novianti, A.N., Hestianah, E.P., Khairullah, A.R. (2024). Effect of garlic extract (Allium sativum) on hematological and clinical chemistry parameters of laying Quail. Asian Journal of Dairy and Food Research. 43(4): 790-795. doi: 10.18805/ ajdfr.DRF-408.

  19. Ievy, S., Hoque, M.N., Islam, M.S. (2022). Genomic characteristics, virulence and antimicrobial resistance in avian pathogenic Escherichia coli MTR_BAU02 strain isolated from layer farm in Bangladesh. Journal of Global Antimicrobial Resistance. 30(1): 155-162. doi: 10.1016j.jgar.2022.06. 001.

  20. Islam, M.S., Nayeem, M.M., Sobur, M.A. (2021). Virulence determinants and multidrug resistance of Escherichia coli isolated from migratory birds. Antibiotics. 10(2): 190. doi: 10.3390/ antibiotics10020190.

  21. Khairullah, A.R., Rehman, S., Sudjarwo, S.A., Effendi, M.H., Ramandinianto, S.C., Gololodo, M.A., Widodo, A., Riwu, K.H.P., Kurniawati, D.A. (2022). Detection of mecA gene and methicillin- resistant Staphylococcus aureus (MRSA) isolated from milk and risk factors from farms in Probolinggo, Indonesia. F1000Research. 11(1): 722. doi: 10.12688/f1000research. 122225.1.

  22. Khairullah, A.R., Sudjarwo, S.A., Effendi, M.H., Kurniawan, S.C., Widodo, A., Silaen, O.S.M., Ramandinianto, S.C. (2024). Identification of Methicillin-resistant Staphylococcus aureus Isolated from Dairy Cow’s Milk in Tulungagung District, Indonesia. Asian Journal of Dairy and Food Research. 43(3): 411-417. doi: 10.18805/ajdfr.DRF-341.

  23. Kusnoto, Yuniarti, W.M., Putri, P.D., Aryaloka, S., Khairullah, A.R., Kurniawan, S.C., Atma, C.D. (2024). Molecular identification of Ascaridia galli in chickens from traditional markets in Surabaya, Indonesia . Asian Journal of Dairy and Food Research. 43(3): 523-528. doi: 10.18805/ajdfr.DRF-379.

  24. Li, J., Fan, Q., Mao, C., Jin, M., Yi, L., Wang, Y. (2021). Effect of biofilm formation on the Escherichia coli drug resistance of isolates from pigs in Central China. Indian Journal of Animal Research. 55(6): 697-703. doi: 10.18805/IJAR.B-1304.

  25. Moses, I.B., Esimone, C.O., Iroha, I.R., Rubin, J.E., Sniatynsky, M.K., Ribeiro, Á.C.D.S., Santos, F.F., da Silva, R.C., Gales, A.C. (2022). Antibiotypes and high frequency of toxin genes in methicillin-resistant Staphylococcus pseudintermedius from nares of dogs and dog guardians in Nigeria. Comparative Immunology, Microbiology and Infectious Diseases. 89(1): 101870. doi: 10.1016/j.cimid.2022.101870.

  26. Moses, I.B., Esimone, C.O., Iroha, I.R., Ugbo, E.N., Orji, J.O., Nwuzo, A.C., Orji, J.O., Nwakaeze, E.A., Agumah, N.B., Emioye, A.A., Ukpai, E.G. and Ogene, L.N. (2020). First report on the antibiotic resistance profiles and virulence genes of Staphylococcus pseudintermedius colonizing dogs and humans in Abakaliki, South-East Nigeria. Research Journal of Microbiology. 15(1): 22-34. doi: 10.3923/jm.2020.22. 34.

  27. Moses, I.B., Ugbo, E.N., Odah, E.E., Iroha, I.R., Agumah, N.B., Ukpai, E.G., Eluu, S.C., Uzoeto, H.O., Okamkpa, C.T., Okata-Nwali, D. (2018). Antibiogram and phenotypic characterization of E. coli Isolated from Nigeria’s paper currencies obtained from butchers in ebonyi State. Archives of Clinical Microbiology. 9(4): 1-5. doi: 10.4172/1989-8436.100085.

  28. Okoli, M.F., Ugwu, M.C., Ezejiegu, K.C. (2021). Antibiotic susceptibility profile of Salmonella spp. and Shigella spp. Isolated from commercial frozen chicken sold in three markets within awka metropolis. Clinical Immunology Research. 5(2): 1-4. 

  29. Pradika, A.Y., Chusniati, S., Purnama, M.T.E., Effendi, M.H., Yudhana, A., Wibawati, P.A. (2019). Total Test of Escherichia coli on fresh cow milk at dairy farmer cooperative (KPSP) Karyo Ngremboko Purwoharjo Banyuwangi. Journal Medik Veteriner. 2(1): 1-6. doi: 10.20473/jmv.vol2.iss1. 2019.1-6.

  30. Putri, M.F.R., Kendek, I.A., Wibisono, F.J., Effendi, M.H., Rahardjo, D., Tyasningsih, W., Ugbo, E.N. (2023). Molecular detection of iron gene on multidrug resistant avian fecal Escherichia coli isolated from broiler on traditional markets, Surabaya, Indonesia. Biodiversitas. 24(12): 6454-6460. doi: 10. 13057/biodiv/d241207.

  31. Rahman, M.M., Husna, A., Elshabrawy, H.A., Alam, J., Runa, N.Y., Badruzzaman, A.T.M., Banu, N.A., Al Mamun, M., Paul, B., Das, S. (2020). Isolation and molecular characterization of multidrug-resistant Escherichia coli from chicken meat. Scientific Reports. 10(1): 21999. doi: 10.1007/s13205- 024-03950-7.

  32. Ranjbar, R., Farahani, A. (2019). Shigella: antibiotic-resistance mechanisms and new horizons for treatment. Infection and Drug Resistance. 12(1): 3137-3167. 

  33. Riwu, K.H.P., Effendi, M.H., Rantam, F.A., Khairullah, A.R., Kurniawan, S.C., Kurniawan, A., Moses, I.B., Hasib, A., Widodo, A., Yanestria, S.M., Silaen, O.S.M., Shehzad, A. (2024). Molecular detection of blaTEM gene for encoding extended spectrum beta-lactamase (ESBL) on Escherichia coli isolated from deer feces in Indonesia. Journal of Advanced Veterinary Research. 14(4): 722-726.

  34. Riwu, K.H.P., Effendi, M.H., Rantam, F.A., Khairullah, A.R., Widodo, A. (2022). A review: Virulence factors of Klebsiella pneumonia as emerging infection on the food chain. Veterinary World. 15(9): 2172-2179. doi: 10.14202/vet world.2022.2172-2179.

  35. Robertson, L.J., Torgerson, P.R., van der Giessen, J. (2018). Food- borne parasitic diseases in Europe: Social cost-benefit analyses of interventions. Trends in Parasitology. 34(11): 919-923. doi: 10.1016/j.pt.2018.05.007.

  36. Roy, K., Islam, M.S., Paul, A. (2022). Molecular detection and anti- biotyping of multi-drug resistant Enterococcus faecium from healthy broiler chickens in Bangladesh. Veterinary Medicine and Science. 8(1): 200-210. doi: 10.1002/vms3. 669.

  37. Sarba, E.J., Kelbesa, K.A., Bayu, M.D., Gebremedhin, E.Z., Borena B.M., Teshale, A.B. (2019). Identification and antimicrobial susceptibility profile of Escherichia coli isolated from backyard chicken in and around ambo, Central Ethiopia. BMC Veterinary Research. 15(1): 85. doi: 10.1186/s12917- 019-1830-z.

  38. Shoja, S., Ghasemi, S., Dastranj, M., Shamseddin, J., Ebrahimi, N., Alizade, H., Farahani, A. (2023). Characterization of genotypes and antimicrobial resistance profiles of clinical isolates of Shigella from patients in the southern region of Iran. European Journal of Medical Research. 28(1): 611. doi: 10.1186/s40001-023-01570-0.

  39. Teimourpour, R., Babapour, B., Esmaelizad, M., Arzanlou, M., Peeri- Doghaheh, H. (2019). Molecular characterization of quinolone resistant Shigella spp. isolates from patients in Ardabil, Iran. Iranian Journal of Microbiology. 11(6): 496. 

  40. Tyasningsih, W., Ramandinianto, S.C., Ansharieta, R., Witaningrum, A.M., Permatasari, D.A., Wardhana, D.K., Effendi, M.H., Ugbo, E.N. (2022). Prevalence and antibiotic resistance of Staphylococcus aureus and Escherichia coli isolated from raw milk in East Java, Indonesia. Veterinary World. 15(8): 2021-2028. doi: 10.14202/vetworld.2022.2021- 2028.

  41. Urmi, M.R., Ansari, W.K., Islam, M.S., Sobur, M.A, Rahman, M., Rahman, M.T. (2021). Antibiotic resistance patterns of Staphylococcus spp. isolated from fast foods sold in different restaurants of Mymensingh, Bangladesh. Journal of Advanced Veterinary and Animal Research. 8(2): 274-281. doi: 10. 5455/javar.2021.h512.

  42. Wareth, G., Neubauer, H. (2021). The Animal-foods-environment interface of Klebsiella pneumoniae in Germany: An observational study on pathogenicity, resistance development and the current situation. Veterinary Research. 52(1): 16. doi: 10.1186/s13567-020-00875-w.

  43. Wibisono, F.J., Sumiarto, B., Untari, T., Effendi, M.H., Permatasari, D.A., Witaningrum, A.M. (2021). Molecular identification of ctx gene of extended spectrum betalactamases (ESBL) producing Escherichia coli on layer chicken in Blitar, Indonesia. Journal of Animal and Plant Sciences. 31(4): 954-959. doi: 10.36899/JAPS.2021.4.0289.

  44. Widodo, A., Lamid, M., Effendi, M.H., Khairullah, A.R., Riwu, K.H.P., Yustinasari, L.R., Kurniawan, S.C., Ansori, A.N.M., Silaen, O.S.M., Dameanti, F.N.A.E.P. (2022). Antibiotic sensitivity profile of multidrug-resistant (MDR) Escherichia coli isolated from dairy cow’s milk in Probolinggo, Indonesia. Biodiversitas. 23(10): 4971-4976. doi: 10.13057/biodiv/ d231002.

  45. Yanestria, S.M., Dameanti, F.N.A.E.P., Musayannah, B.G., Pratama, J.W.A., Witaningrum, A.M., Effendi, M.H., Ugbo, E.N. (2022). Antibiotic resistance pattern of Extended-Spectrum â- Lactamase (ESBL) producing Escherichia coli isolated from broiler farm environment in Pasuruan district, Indonesia. Biodiversitas. 23(9): 4460-4465. doi: 10.13057/biodiv/ d230911.

  46. Yanestria, S.M., Effendi, M.H., Tyasningsih, W., Moses, I.B., Khairullah, A.R., Kurniawan, S.C., Dameanti, F.N.A.E.P., Ikaratri, R., Pratama, J.W.A., Sigit, M., Hasib, A., Silaen, O.S.M. (2024). Antimicrobial resistance patterns and genes of Campylobacter jejuni isolated from chickens in Pasuruan, Indonesia. Open Veterinary Journal. 14(3): 759-768. doi: 10.5455/ OVJ.2024.v14.i3.2.

  47. Yanestria, S.M., Rahmaniar, R.P., Wibisono, F.J., Effendi, M.H. (2019). Detection of invA gene of Salmonella from milkfish (Chanos chanos) at Sidoarjo wet fish market, Indonesia, using polymerase chain reaction technique. Veterinary World. 12(1): 170-175. doi: 10.14202/vetworld.2019.170-175.
Reviewed By

Download Article
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)