EJBR2020v10i1art1 ISSN 2449-8955 European Journal of Biological Research Research Article European Journal of Biological Research 2020; 10(1): 1-10 DOI: http://dx.doi.org/10.5281/zenodo.3630802 Pseudomonas species from cattle dung producing extended spectrum and metallo beta-lactamases Olutayo Israel Falodun*, Isaiah Baba Musa Department of Microbiology, University of Ibadan, Ibadan, Nigeria *Correspondence: Phone: +2348027342286; E-mail: falod2013@gmail.com Received: 10 November 2019; Revised submission: 12 December 2019; Accepted: 29 January 2020 http://www.journals.tmkarpinski.com/index.php/ejbr Copyright: © The Author(s) 2019. Licensee Joanna Bródka, Poland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/) ABSTRACT: Indiscriminate use of antibiotics in livestock contributes to emergence of antimicrobial resistance in pathogens co-habiting the gastro-intestinal tract of animals. This study was to determine the Extended Spectrum Beta-Lactamase (ESBL) and Metallo-Beta-Lactamase (MBL) production in Pseudomonas species from cattle fecal samples. Cattle dungs were collected from the University of Ibadan Cattle Ranch and the Pseudomonas species isolated using Pseudomonas Base Agar with Pseudomonas CN Selective Supplement were identified using standard tests. Phenotypic detection of ESBL and MBL was by double disk synergy test and Ethylene Di-amine Tetra Acetic Acid Combined Disk Test respectively. Antibiotics susceptibility tests was done using the disc diffusion technique against ten antibiotics. A total of 144 Pseudomonas species were isolated and identified as P. aeruginosa (71.5%), P. fluorescens (19.4%) and P. stutzeri (9.1%) and 19 (37.1%) produced ESBL including P. aeruginosa (15), P. fluorescens (2) and P. stutzeri (2) while, one (6.7%) ESBL P. aeruginosa produced MBL. All the ESBL producers were resistant to cefotaxime and trimethoprim; resistance of P. aeruginosa to ciprofloxacin was 93.3% and to ceftazidime was 80.0%, while it was 13.3% (colistin) and 6.7% (imipenem). The ESBL producing P. fluorescens were resistant to ceftazidime, ciprofloxacin and trimethoprim, likewise, the ESBL producing P. stutzeri showed resistance to gentamicin, ciprofloxacin and trimethoprim. The production of ESBL and MBL observed among the Pseudomonas species in this study with high level of resistance to some antibiotics portend public health risk, hence a need for caution in the use of antibiotics in animal husbandry. Keywords: Pseudomonas species; Cattle dung; ESBL; MBL; Antibiotic resistance. 1. INTRODUCTION In other to improve animal health and productivity especially in intensive reared species in agricultural industry, the use of antimicrobial agents is relied upon [1, 2]. The indiscriminate use of antimicrobial agents in livestock management either as food supplements or growth promoters has led to the emergence of Extended Spectrum Beta-lactamases (ESBLs) and Metallo-Beta-Lactamases (MBLs) producing bacteria including opportunistic pathogens such as Pseudomonas species of the gastrointestinal tract owing to mutations, selective pressure and the widespread of multi-drug resistance genes amongst bacteria [3]. The genus Pseudomonas is one of the most important members of the family Pseudomonadaceae which are Gram- negative bacilli, aerobic gamma-Proteobacteria with straight or sometimes marginally bent rod shape and one Falodun & Musa Beta-lactamases Pseudomonas in cattle 2 European Journal of Biological Research 2020; 10(1): 1-10 or more polar flagella [4]. The use of antibiotics in the livestock production chain is usually seen as important in continuing a consistent supply of healthy and substantial animals, leading to greater profitability and efficiency [5]. Interestingly, many of the antimicrobial agents such as tetracycline, penicillin and sulphonamides that are important in human health are also used in animal food production [6]. It has also been observed that the application of mass medication known as metaphylaxis and the use of broad-spectrum antibiotics in animal husbandry especially for nontherapeutic use is linked to resistance in people who live on and near farms and even the general population via the food chain [7]. Extended Spectrum Beta-lactamases (ESBLs) are plasmid mediated beta-lactamases that mediate resistance to Extended Spectrum Cephalosporins (ESCs) including ceftriaxone, ceftazidime and cefotaxime and the monobactam such as aztreonam but have no effect on cephamycins and carbapenems. The ESBLs hydrolyzes the oxyimino-cephalosporins by cleaving structural beta-lactam ring but are inhibited by beta lactamase inhibitors such as clavulanic acid, tazobactam and sulbactam [8]. Extended Spectrum Beta- lactamases have been increasingly reported to be produced by the members of family Enterobacteriaceae and by Pseudomonas species [9]. Metallo-Beta-Lactamases (MBLs) is enzymes capable of hydrolysing bicyclic beta-lactam antibiotics such as penicillins, cephalosporins and carbapenems with the exception of the monobactams. Metallo-Beta-Lactamases producing Pseudomonas species was first reported in Japan in 1991 and since then, there had been substantial increase in Pseudomonas producing MBLs worldwide [10]. The association of MBLs genes to mobile genetic elements has facilitated the dissemination of these enzymes among prevalent pathogens thereby, increasing the spread and outbreak of community and hospital acquired infection [10, 11]. This study was designed to determine the occurrence of ESBLs and MBLs production in Pseudomonas species isolated from cattle dung collected from the cattle ranch of the University of Ibadan, Nigeria and determine their antibiotic susceptibility pattern. 2. MATERIALS AND METHOD 2.1. Study site and sample collection The study site was the University of Ibadan Cattle Ranch located in Abadina end of the University. A total of thirty (30) cattle dung samples were collected from the Ranch. The sample bottles were labelled appropriately, placed in ice packs and immediately transported to the Microbiology laboratory of the University of Ibadan for processing. 2.2. Isolation and identification of the Pseudomonas species Pseudomonas species were isolated using the method of Kathiravan et al. [12]. Pseudomonas base agar (CM0559, Oxoid) supplemented with Pseudomonas C-N supplement (SR102, Oxoid), a selective media, prepared according to the manufacturers’ instruction was used for the isolation of the Pseudomonas spp. One ml of the serial diluents (10-1) of the samples was dispensed into appropriately labelled Petri dishes. Aseptically, Pseudomonas base agar cooled to about 45oC was dispensed into the aliquots of the samples in the Petri dishes and swirled gently, allowed to solidified and incubated at 37oC for 24-48 hours [12]. The isolates were characterized using standard morphological and biochemical tests including Gram staining, catalase, motility, oxidase, growth at 4oC, growth at 42oC and sugar fermentation tests including glucose, lactose, maltose, mannitol, sucrose, nitrate reduction, citrate [13]. Falodun & Musa Beta-lactamases Pseudomonas in cattle 3 European Journal of Biological Research 2020; 10(1): 1-10 2.3. Screening for potential Extended Spectrum Beta-Lactamases (ESBL) producing Pseudomonas species All the isolates were subjected to antibiotics susceptibility testing using Kirby-Bauer disk diffusion test. The antibiotics discs used were ceftazidime (30 μg), cefotaxime (30 μg) and cefepime (30 μg) purchased from Oxoid, UK. Pure distinct colonies of 18-24 hours old culture were inoculated into sterile test tubes containing normal saline and its turbidity was adjusted to 0.5 McFarland standards. Mueller Hinton agar plates were prepared according to manufacturer’s instructions and the standardized bacterial suspension was evenly inoculated on the surface of the agar plate by swabbing the entire surface of the agar. The antibiotics were placed on the culture plates with the aid of a sterile forceps and incubated at 37oC for 18-24 hours. The diameters of the zones of inhibitions were measured, recorded in mm and interpreted using Clinical Laboratory Standard Institute (CLSI) and those with reduced susceptibility were selected as potential ESBL- producers [14]. 2.4. Phenotypic detection of Extended Spectrum Beta-Lactamases (ESBL) producing Pseudomonas species using Double Disk Synergy Test (DDST) All the isolates that showed reduced susceptibility to ceftazidime (30 μg), cefotaxime (30 μg) and cefepime (30 μg) were selected for ESBL detection using double disk synergy test. This was done using disks of ceftazidime (30 μg), cefotaxime (30 μg) and cefepime (30 μg); which were placed adjacent to augmentin (amoxicillin-clavulanate 20 μg/10 μg) disk at the distance of 20 mm from it (centre to centre). The standardized bacterial test suspension was inoculated on Mueller Hinton agar plates by uniformly swabbing the entire surface of the agar plates and the inoculated plates were incubated for 18-24 hours at 37oC. Isolates producing ESBL were those with a clear cut indentation towards the amoxicillin-clavulanate disc [15]. 2.5. Screening for potential Metallo-Beta-Lactamase (MBL) producing Pseudomonas species Phenotypic screening of the isolates for MBL production was carried out by subjecting the isolates that produced ESBL to imipinem (30 μg) purchased from Oxoid (UK), using Kirby-Bauer disk diffusion test. Pure distinct colonies of 18-24 hours old culture of the isolates were inoculated into sterile test tubes containing normal saline and turbidity adjusted to 0.5 McFarland standards. The standardized bacterial suspension was evenly inoculated on the entire surface of Mueller Hinton agar plates by swabbing. The antibiotics were placed on the culture plates with the aid of a sterile forceps and incubated at 37oC for 18-24 hours. Isolates that showed inhibition zone diameter (IZD) of ≤ 23 mm were considered and suspected to produce MBL enzyme and these isolates were further tested using a phenotypic confirmation test [14]. 2.6. Phenotypic detection of Metallo-Beta-lactamase (MBL) producing Pseudomonas species The metallo-beta-lactamase production of the isolates was determined phenotypically using Ethylene Di-amine Tetra Acetic Acid (EDTA) Combined Disk Test (EDTA-CDT). One disk of imipenem (10 μg) and one with imipenem (10 μg) in combination with 0.5 M EDTA were placed at a distance of 20 mm, centre to centre, on Mueller Hinton agar plate inoculated with a bacterial suspension of 0.5 McFarland turbidity and incubated at 37oC for 18-24 hours. The MBL producers were those with zone of inhibition with a difference of 7 mm and above around imipenem disk containing EDTA compared to imipenem disk without EDTA [16]. Falodun & Musa Beta-lactamases Pseudomonas in cattle 4 European Journal of Biological Research 2020; 10(1): 1-10 2.7. Antibiotics susceptibility tests of the ESBL producing Pseudomonas species Antibiotics susceptibility test of the ESBL-producing Pseudomonas species were carried out using the standard disk diffusion method recommended by Clinical Laboratory Standard Institute against ceftazidime (30 μg), cefotaxime (30 μg), cefepime (30 μg), amoxicillin-clavulanate (20 μg/10 μg), gentamicin (10 μg), ciprofloxacin (5 μg), Imipenem (10 μg), colistin (10 μg), trimethoprim (5 μg) and aztreonam (30 μg). The susceptibility test was carried out using pure colonies of 18-24 hours old culture adjusted to 0.5 McFarland Standards. The culture suspension was inoculated unto the surface of Mueller Hinton agar plates using sterile swab sticks. The antibiotics discs were placed on the inoculated plates with the aid of a sterile forceps and incubated at 37oC for 18-24 hours. The zones of inhibition were measured and interpreted according to Clinical Laboratory Standard Institute [14]. 3. RESULTS The average mean value of the total heterotrophic bacteria count obtained from the cattle dung was 2.3×10-6 cfu/g, with the highest mean value of 2.7×10-6 cfu/g from paddock 4 and the least (1.8×10-6cfu/g) from paddock 1 (Table 1). A total of 144 Pseudomonas species were isolated including P. aeruginosa (71.5%), P. fluorescens (19.4%) and P. stutzeri (9.1%) (Table 2). Of the 144 Pseudomonas species, 19 (37.1%) were positive for ESBL production, comprising 15 (14.6%) P. aeruginosa, 2 (7.1%) P. fluorescens and 2 (15.4%) P. stutzeri (Table 2). In addition, only 1 (5.3%) Pseudomonas aeruginosa that produced ESBL also produced MBL (Table 2). Table 1. Total Heterotrophic Bacteria Count (THBC) of isolates from the cattle dung. Sum of THBC (×10-4CFU/g) Mean ± SD Paddock 1 544 181.3 ± 17.0 Paddock 2 734 244.7 ± 31.1 Paddock 3 790 263.3 ± 19.4 Paddock 4 819 273.0 ± 17.7 Paddock 5 576 192.0 ± 13.1 Paddock 6 723 241.0 ± 24.0 Paddock 7 796 265.3 ± 13.0 Paddock 8 703 234.2 ± 20.5 Paddock 9 759 253.0 ± 8.2 Paddock 10 550 183.3 ± 15.0 Total 6994 233.1 ± 16.1 Table 2. Occurrence of Extended Spectrum Beta-Lactamases (ESBLs) and Metallo Beta-Lactamases (MBL) producing Pseudomonas species from cattle dung. Isolates No. of tested isolates n (%) of positive ESBL n (%) of positive MBL P. aeruginosa 103 15 (14.6) 1 (6.7) P. fluorescens 28 2 (7.1) 0 (0) P. stutzeri 13 2 (15.4) 0 (0) Total 144 19 (13.2) 1 (5.3) The patterns of the antimicrobial resistance of the ESBL isolates showed that all the 19 (100%) isolates were resistant to trimethoprim and cefotaxime. Of the 15 P. aeruginosa that produced ESBL, 12 (80.0%) Falodun & Musa Beta-lactamases Pseudomonas in cattle 5 European Journal of Biological Research 2020; 10(1): 1-10 showed resistance to ceftazidime, 9 (60.0%) to gentamicin while, 1 (5.3%) and 2 (13.3%) were resistant to imipenem and colistin respectively. However, none of these isolates showed resistance to cefepime and aztreonam. Furthermore, the two P. fluorescens that produced ESBL also showed resistance to ciprofloxacin and ceftazidime. Similarly, the two P. stutzeri ESBL producers showed resistance to gentamicin and ciprofloxacin, but the two P. fluorescens and P. stutzeri were fully susceptible to cefepime, aztreonam, imipenem and colistin (Table 3). In addition, all the ESBL producers showed resistance to at least four different classes of antibiotics. Five (26.5%) of the isolates showed resistance to a combination of four antibiotics (CAZ-CTX-CIP-SXT) including four P. aeruginosa and one P. fluorescens while two isolates including one each of the P. aeruginosa and one P. fluorescens showed resistance to a combination of six (AMC-CAZ-CTX-CIP-GEN-SXT) antibiotics and one P. aeruginosa also showed resistance to eight (AMC-CAZ-CTX-CIP-GEN-CST-IPM- SXT) antibiotics (Table 4). Table 3. Antibiotics resistant pattern of the ESBL producing Pseudomonas species isolated from the cattle dung. Antibiotics P. aeruginosa n=15 (%) P. fluorescens n=2 (%) P. stutzeri n=2 (%) Amoxicillin-clavulanate 6 (40) 1 (50) 0 (0) Ceftazidime 12 (80) 2 (100) 1 (50) Cefotaxime 15 (100) 2 (100) 2 (100) Cefepime 0 (0) 0 (0) 0 (0) Aztreonam 0 (0) 0 (0) 0 (0) Imipenem 1 (6.7) 0 (0) 0 (0) Gentamicin 9 (60) 1 (50) 2 (100) Colistin 2 (13.3) 0 (0) 0 (0) Ciprofloxacin 14 (93.3) 2 (100) 2 (100) Trimethoprim 15 (100) 2 (100) 2 (100) Table 4. Antibiotypes of the Extended Spectrum Beta-Lactamases (ESBLs) producing Pseudomonas species from the cattle dung. Antibiotypes Pseudomonas aeruginosa n=15 Pseudomonas fluorescens n=2 Pseudomonas stutzeri n=2 Total n=19 CAZ-CTX-CIP-SXT 4 (26.7%) 1 (50%) - 5 (26.5%) CAZ-CTX-GEN-SXT 2 (13.3%) - - 2 (10.5%) AMC-CTX-CIP-SXT 1 (6.7%) - - 1 (5.3%) CTX-CIP-GEN-SXT 0 (0%) - 1 (50%) 1 (5.3%) CAZ-CTX-CIP-GEN-SXT 3 (20%) - - 3 (15.8%) AMC-CAZ-CTX-CIP-SXT 2 (13.3%) - - 2 (10.5%) CAZ-CTX-CIP-GEN-SXT 0 (0%) - 1 (50%) 1 (5.3%) AMC-CAZ-CTX-CIP-GEN-SXT 1 (6.7%) 1 (50%) - 2 (10.5%) AMC-CAZ-CTX-CIP-GEN-CST-SXT 1 (6.7%) - - 1 (5.3%) AMC-CAZ-CTX-CIP-GEN-CST-IPM-SXT 1 (6.7%) - - 1 (5.3%) Footnote: CAZ: Ceftazidime; CTX: Cefotaxime; FEP: Cefepime; AMC: Amoxicillin-clavulanate; CIP: Ciprofloxacin; CST: Colistin; GEN: Gentamicin; SXT: Trimethoprim; IMP: Imipenem. Falodun & Musa Beta-lactamases Pseudomonas in cattle 6 European Journal of Biological Research 2020; 10(1): 1-10 4. DISCUSSION The average mean value of the total heterotrophic bacterial count (THBC) (2.3×106 cfu/g) observed in this study is similar to the average mean value of 2.71×106 cfu/g reported from a previous study on cattle’s faecal sample from an abattoir in Gombe State, northern part of Nigeria [17]. The observed highest mean value (2.7×106 cfu/g) in this study is not in agreement with 8.65×107 cfu/g from cow dung in Cross River, a city in the southern part of Nigeria [18]. The disparity might be due to differences in the plating techniques, while pour plate technique was employ in this study, spread plate technique was used in the latter study. Similarly, the least mean value (1.8×106 cfu/g) from this study is lower compared to 2.29×108 cfu/ml reported from another study on cow dung in India [19]. The difference might be due to the geographical locations. The high THBC mean value obtained from the cattle dung in each Paddock revealed the presence of high microbial load and a similar report attributed this to rich microbial diversity of animal guts [20]. The prevalence P. aeruginosa (71.5%) in this study is not in agreement with the previously reported 30.0% obtained in a similar study in Ebonyi, Southeastern Nigeria [21]. Pseudomonas species had been predominantly reported to be found on plant and water bodies and the animals from the present study were allowed to freely graze on open field pasture (grasses) surrounding the ranch with their water source from a nearby river [22]. In addition, the prevalence (9.1%) of P. stutzeri obtained from the present study was a bit higher than the 2.1% recently reported from a study carried out on fishes in Uganda [23]. The reason for the differences might be the studied samples and the fact that the source of drinking water for the animals in the present study is a nearby river, and aquatic environment had been reported to be potential reservoirs for Pseudomonas species [24, 25]. Furthermore, the 37.1% ESBL producing Pseudomonas species in this study is comparably similar to the 38.9% from a recent study on Pseudomonas species isolated from selected rivers in Ibadan [26]. This finding is also similar to 37.8% ESBL production from a study in Bangladesh on human clinical samples [27]. However, a much lower occurrence (15.0%) was reported in a study carried out on Pseudomonas species from mixed human samples in Enugu, Nigeria [28]. The reason for this disparity could be due the number of isolates studied. While 144 isolates were used in this study, only 20 isolates were studied in the latter research. More so, animals from which samples were collected from the present study were pre-exposed to treatment with various antibiotics including the beta-lactams classes and previous report had attributed high prevalence of ESBL producing Pseudomonas species in animal husbandry to the wide misuse and abuse of antibiotics [29]. In addition, the prevalence (6.7%) of ESBL and MBL co-production among the P. aeruginosa is comparably similar to the 5.1% previously reported from another study carried out on P. aeruginosa isolated from human clinical samples in India [30]. However, this observation is slightly higher than the 2.8% and 3.3% reported from studies on P. aeruginosa on clinical samples in France and Abeokuta, Nigeria respectively [31, 32]; but lower compared to the 18.6% previously reported in a similar study in Abakaliki, Nigeria [21]. The coexistence of ESBLs and MBLs enzymes in a single P. aeruginosa poses a public health risk to mankind owing to the fact that these genes are reported to be plasmid encoded and could be transferred from one organisms to another within the guts, thus conferring resistance to antimicrobial agents such as aminoglycosides, macrolides, carbapenems and sulphamethoxazole [33, 34]. Furthermore, the total resistance of the ESBL-producing Pseudomonas species observed in this study to trimethoprim is in agreement with the report of studies on commensal Pseudomonas species from wastewater and freshwater Milieus in the Eastern Cape Province, South Africa and human clinical samples in Iran [25, 34]. However, this observation is not in agreement with the 38.0% resistance reported on similar Falodun & Musa Beta-lactamases Pseudomonas in cattle 7 European Journal of Biological Research 2020; 10(1): 1-10 isolates from camel in Egypt [35]. The discrepancy might be due to the differences in sampling source. In addition, the observed total resistance of ESBL-producing P. fluorescens and P. stutzeri and high (93.3%) resistance of P. aeruginosa to ciprofloxacin in this study contradict the report of other studies on abattoir wastewater in Ibadan, Nigeria and camels samples in Egypt from which none of the isolates and 33.3% Pseudomonas aeruginosa showed resistance to ciprofloxacin respectively [36, 37]. The reason for the high level of resistance in the present study may be due to the indiscriminate use of these classes of antibiotics in livestock management that might have led to selective pressure and development of resistance. It has also been revealed in previous report that the use of antibiotics as supplement in commercial feeds and as growth enhancers might have initiated resistance [38]. In addition, the mechanisms for resistance employ by P. aeruginosa to quinolones include: decreased in the amount of quinolones entering the cell because of the defect in the function of the porin channels and various efflux systems in the bacterial membrane [39]. The 6.7% resistance to imipenem among the ESBL-producing P. aeruginosa in this study is in agreement with the 6.0% previously reported on P. aeruginosa isolated from human blood, wound, sputum, cerebral spinal fluid, stool, ear and eye swab in Tehran [40]. However, that none of the ESBL-producing P. fluorescens and P. stutzeri showed resistance to imipenem is not in agreement with 6.7% that was obtained in another study carried out on ESBL producing Pseudomonas species isolated from human wounds, pus, urine aural sputum, throat umbilicus and conjunctiva samples [27]. The low resistance to imipenem may be because it is not readily available for the treatment of animal infections. Because the carbapenems such as imipenem are considered as last resort for treatment, there is a need for continued surveillance and judicious use of these antibiotics especially in livestock management [21]. Moreover, the 13.3% resistance to colistin by ESBL-producing P. aeruginosa is a public health challenge because colistin is regarded as one of the drug of choice and last drug of resort for the treatment of infections caused by multidrug resistance P. aeruginosa. The implication of this is that infections caused by these organisms may be difficult to treat. Furthermore, the observation that none of the ESBL-producing P. fluorescens and P. stutzeri showed resistance to cefepime, aztreonam and colistin is in contrast with the report of Chen et al. [41] in a study where the ESBL producing P. aeruginosa isolated from clinical specimens in Chinese Teaching Hospital showed a higher resistance (52.4%) to both cefepime and aztreonam. The reason for the differences may be due to samples studied. The observation from this study that showed the ESBL-producing P. aeruginosa exhibiting multiple drug resistance to a combination of seven (7) different classes of antibiotics (AMC-CAZ-CTX-CIP-GEN- CST-IPM-SXT) is similar and comparable to a study carried on P. aeruginosa isolated from wastewater generated from an abattoir in Ibadan, Nigeria which showed resistance to a combination of 8 different classes of antibiotics (AMP-TET-CHL-CRO-OFX-CLX-STR-SXT) but higher than the one reported in another study on commensal Pseudomonas species from wastewater and fresh water milieus in South Africa which showed resistance to the combination of four (4) different classes of antibiotics (PG-OX-CD-RP) [25, 36]. The observed resistance in the strains of Pseudomonas species obtained from cattle is alarming as these animals could serve as potential reservoirs of these resistant strains and could be deposited into the environment in the form feces/urine often used as manure on crop produce. These bacterial strains could be transmitted directly and indirectly to human because cattle meat are frequently consumed in Nigeria as part of diet in a roasted or cooked form and thus, posing serious health threat when such meat product harboring these pathogens are not properly cooked [22]. The resistance patterns of ESBL-producing Pseudomonas species against the various antibiotics tested in the present study showed that all the isolates obtained from cattle dung were multidrug resistant as they showed resistance to a combination of three or more different classes of antibiotics. The ability of Falodun & Musa Beta-lactamases Pseudomonas in cattle 8 European Journal of Biological Research 2020; 10(1): 1-10 Pseudomonas species to acquire and harbour various resistance determinants allows only limited classes of antibiotics for effective treatment of infection caused by it. 5. CONCLUSION The ESBL and MBL producing Pseudomonas species in this study showed high level of resistance to some commonly available antibiotics especially the beta-lactams. The emergence and spread of these bacteria in cattle might be as a result of the indiscriminate use of these antibiotics and intrinsic resistance properties which could portend public health risk to mankind and the environment. Hence, the use of antibiotics in animal husbandry should be regulated. Authors’ Contributions: OIF designed the study and protocol, supervised the study, managed literature search, data acquisition and analysis, revised the manuscript. 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