Protection of Chickens against Escherichia coli Infections by DNA Containing CpG Motifs

ABSTRACT Synthetic oligodeoxynucleotides (ODN) containing CpG motifs (CpG-ODN) have been shown to be effective immunoprotective agents in murine models for a variety of viral, intracellular bacterial, and protozoan infections. Until now, the use of CpG-ODN to protect against extracellular bacterial infections has not been reported. The objective of this study was to investigate the effect of CpG-ODN against cellulitis and colibacillosis in broiler chickens, using a well-established model. At 22 days of age, birds received CpG-ODN by either the subcutaneous or intramuscular route. Three days later, a virulent isolate of Escherichia coli was applied to a scratch site on the caudal abdominal skin. Birds were examined for 10 days after the E. coli challenge, and pathological and bacteriological assessments were conducted on all birds. The control group of birds receiving no CpG-ODN(2007) had a survival rate of 15%. In contrast, groups that received CpG-ODN(2007), by either subcutaneous or intramuscular injection, had significantly higher survival rates (P < 0.0001). Furthermore, the size of the cellulitis lesion was significantly smaller in groups that received CpG-ODN(2007) by the subcutaneous route (P < 0.01). A dose of as little as 3.16 μg of CpG-ODN(2007), delivered 3 days prior to challenge by either the subcutaneous or intramuscular route, significantly protected birds against E. coli infection (P < 0.01). This study demonstrates that CpG-ODN(2007) has both local and systemic protective effects in broiler chickens. This is the first time that CpG-ODN(2007) has been demonstrated to have an immunoprotective effect against an extracellular bacterial infection in any food animal species.

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Interleukin gene expression in broiler chickens infected by different Escherichia coli serotypes

Veterinary World ◽

10.14202/vetworld.2021.2727-2734 ◽

2021 ◽

pp. 2727-2734

Author(s):

Reham Elnagar ◽

Rasha Elkenany ◽

Gamal Younis

Keyword(s):

Gene Expression ◽

Escherichia Coli ◽

Broiler Chickens ◽

Quantitative Polymerase Chain Reaction ◽

Clinical Signs ◽

Negative Control Group ◽

Negative Control ◽

Control Group ◽

E Coli ◽

High Prevalence

Background and Aim: Escherichia coli is the cause of avian colibacillosis, a significant threat to the poultry industry and public health. Thus, this study investigated the prevalence of E. coli in diseased chicken broilers, pathological effects of these bacteria, and interleukin (IL) gene expression of different serotypes of E. coli (O78, O26, O44, and O55) on experimentally infected chickens. Materials and Methods: A total of 295 organ samples (liver, lungs, heart, and spleen) from 59 diseased broiler chickens were used for conventional identification of E. coli. Chickens were orally infected with one of the following E. coli serotypes (O78, O26, O44, or O55) and examined for clinical signs, mortality, macroscopic and microscopic lesions, and IL gene expression using real-time quantitative polymerase chain reaction. Results: E. coli was isolated from 53.2% of broiler chicken organs with a high prevalence in lungs (26.1%). The most prevalent serotypes were O78, O26, O44, O55, O157, and O127 prevalence of 27.8, 22.2, 16.7, 16.7, 5.6, and 5.6%, respectively. In the experimental design, five groups (G1-G5) of birds were established. G1 served as the negative control group, while G2-G5 were challenged orally with E. coli O78, O26, O55, or O44, respectively. Chickens infected with E. coli O78 or O26 showed significant clinical signs in comparison to the other infected birds. Mortality (13.3%) was only observed in birds infected with E. coli O78. Necropsy of dead birds after E. coli O78 infection showed pericarditis, enteritis, airsacculitis, and liver and lung congestion. More severe histopathological changes were observed in intestines, spleen, liver, and lung from chickens infected with either E. coli O78 or O26 than for birds infected with other serotypes. On the 2nd day post-infection, E. coli challenge, particularly with E. coli O78, displayed significantly upregulated levels of ileal IL-6 and IL-8, but ileal IL-10 level tended to be downregulated in comparison to the control group. Conclusion: This study assessed the application of cytokines as therapeutic agents against infectious diseases, particularly colibacillosis.

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Effects of In-Feed Chlortetracycline Prophylaxis in Beef Cattle on Animal Health and Antimicrobial-Resistant Escherichia coli

Applied and Environmental Microbiology ◽

10.1128/aem.01928-16 ◽

2016 ◽

Vol 82 (24) ◽

pp. 7197-7204 ◽

Cited By ~ 21

Author(s):

Getahun E. Agga ◽

John W. Schmidt ◽

Terrance M. Arthur

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Human Health ◽

Bacterial Infections ◽

Animal Health ◽

Agricultural Practices ◽

Control Group ◽

Content Type ◽

E Coli ◽

The Impact

ABSTRACTConcerns have been raised that in-feed chlortetracycline (CTC) may increase antimicrobial resistance (AMR), specifically tetracycline-resistant (TETr)Escherichia coliand third-generation cephalosporin-resistant (3GCr)E. coli. We evaluated the impact of a 5-day in-feed CTC prophylaxis on animal health, TETrE. coli, and 3GCrE. coli. A control group of cattle (n= 150) received no CTC, while a CTC group (n= 150) received in-feed CTC (10 mg/lb of body weight/day) from the 5th to the 9th day after feedlot arrival. Over 25% (38/150) of the animals in the control group developed illnesses requiring therapeutic treatment with antimicrobials critically important to human medicine. Only two animals (1.3%) in the CTC group required such treatments. Fecal swab and pen surface occurrences of genericE. coli(isolated on media that did not contain antimicrobials of interest and were not isolated based on any specific resistance), TETrE. coli, and 3GCrE. coliwere determined on five sampling occasions: arrival at the feedlot, 5 days posttreatment (5 dpt), 27 dpt, 75 dpt, and 117 dpt. On 5 dpt, TETrE. coliconcentrations were higher for the CTC group than the control group (P< 0.01). On 27 dpt, 75 dpt, and 117 dpt, TETrE. coliconcentrations did not differ between groups. 3GCrE. colioccurrences did not differ between control and CTC groups on any sampling occasion. For both groups, generic, TETr, and 3GCrE. colioccurrences were highest on 75 dpt and 117 dpt, suggesting that factors other than in-feed CTC contributed more significantly to antimicrobial-resistantE. colioccurrence.IMPORTANCEThe occurrence of human bacterial infections resistant to antimicrobial therapy has been increasing. It has been postulated that antimicrobial resistance was inevitable, but the life span of the antimicrobial era has been prematurely compromised due to the misuse of antimicrobials in clinical and agricultural practices. Direct evidence relating the use of antimicrobials in livestock production to diminished human health outcomes due to antimicrobial resistance is lacking, and the U.S. Food and Drug Administration has taken an approach to maximize therapeutic efficacy and minimize the selection of resistant microorganisms through judicious use of antimicrobials. This study demonstrated that prophylactic in-feed treatment of chlortetracycline administered for 5 days to calves entering feedlots is judicious, as this therapy reduced animal morbidity, reduced the use of antimicrobials more critical to human health, and had no long-term impact on the occurrence of antimicrobial-resistantE. coli.

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Critical Roles of Spätzle5 in Antimicrobial Peptide Production Against Escherichia coli in Tenebrio molitor Malpighian Tubules

Frontiers in Immunology ◽

10.3389/fimmu.2021.760475 ◽

2021 ◽

Vol 12 ◽

Author(s):

Maryam Ali Mohammadie Kojour ◽

Tariku Tesfaye Edosa ◽

Ho Am Jang ◽

Maryam Keshavarz ◽

Yong Hun Jo ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Peptide ◽

Bacterial Infections ◽

Tenebrio Molitor ◽

Larval Survival ◽

Malpighian Tubules ◽

Control Group ◽

Mrna Levels ◽

E Coli ◽

Pathway Activation

The dimeric cytokine ligand Spätzle (Spz) is responsible for Toll pathway activation and antimicrobial peptide (AMP) production upon pathogen challenge in Tenebrio molitor. Here, we indicated that TmSpz5 has a functional role in response to bacterial infections. We showed that the highest expression of TmSpz5 is induced by Candida albicans. However, TmSpz5 knockdown reduced larval survival against Escherichia coli and Staphylococcus aureus. To evaluate the molecular mechanism underlying the observed survival differences, the role of TmSpz5 in AMP production was examined by RNA interference and microbial injection. T. molitor AMPs that are active against Gram-negative and -positive bacteria, including Tmtenecins, Tmattacins, Tmcoleoptericins, Tmtaumatin-like-proteins, and Tmcecropin-2, were significantly downregulated by TmSpz-5 RNAi in the Malpighian tubules (MTs) following a challenge with E. coli and S. aureus. However, upon infection with C. albicans the mRNA levels of most AMPs in the dsTmSpz5-injected group were similar to those in the control groups. Likewise, the expression of the transcription factors NF-κB, TmDorX2, and TmRelish were noticeably suppressed in the MTs of TmSpz5-silenced larvae. Moreover, E. coli-infected TmSpz5 knockdown larvae showed decreased antimicrobial activity in the MTs and hindgut compared with the control group. These results demonstrate that TmSpz5 has a defined role in T. molitor innate immunity by regulating AMP expression in MTs in response to E. coli.

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Evaluation of commercially available aroA delated gene E. coli O78 vaccine in commercial broiler chickens under Middle East simulating field conditions

Scientific Reports ◽

10.1038/s41598-021-81523-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Hussein M. Galal ◽

M. I. Abdrabou ◽

Ahmed H. I. Faraag ◽

C. K. Mah ◽

Azza M. Tawfek

Keyword(s):

Middle East ◽

Broiler Chickens ◽

Bacterial Infections ◽

Body Weight Gain ◽

Broiler Chicks ◽

E Coli ◽

Reduced Body ◽

Antibiotics Use ◽

Commercial Broiler ◽

And Performance

AbstractThe broiler industry in the Middle East (ME) faces many challenges related to bacterial infections, including M. gallisepticum, M. synoviae, E. coli, and other gram-negative bacteria, exacerbated by various errors in the brooding process. Antibiotics use in the first three days of life, such as Linco-Spectin 100 SP, tilmicosin, enrofloxacin, tylosin, colistin, and doxycycline, is the trend in the market to control such challenges. This study aimed to evaluate the efficacy of the newly introduced aroA E. coli vaccine (Poulvac E. coli) and its ability to reduce over-reliance on the heavy use of antibiotics in the ME. The study was conducted on 160 broiler chicks, divided into eight even groups. Each group was treated differently in terms of antibiotic therapy and ages at the time of Poulvac E. coli administration and the challenge of virulent avian pathogenic E. coli (APEC), serotype O78. Spray application of Poulvac E. coli at seven days of age plus Linco-Spectin 100 SP during the first three days provided the best results for zero mortality after challenge with APEC, while Poulvac E. coli at seven days with enrofloxacin during the early three days resulted in 10% mortality. Poulvac E. coli hatchery vaccination protected birds against mortality but reduced body weight gain compared to the 7-day group vaccinated with Linco-Spectin 100 SP during the first three days. Poulvac E. coli given on day one or day seven did not affect the immune response to concurrent respiratory viral vaccines and, in some cases, improved response. This study shows that Poulvac E. coli at seven days of age, together with Linco-Spectin 100 during the first three days, has produced the best results in terms of protection and performance in the ME high presence of avian pathogenic E. coli field challenge.

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Three-Year Trend in Escherichia coli Antimicrobial Resistance among Children’s Urine Cultures in an Italian Metropolitan Area

Children ◽

10.3390/children8070597 ◽

2021 ◽

Vol 8 (7) ◽

pp. 597

Author(s):

Luca Pierantoni ◽

Laura Andreozzi ◽

Simone Ambretti ◽

Arianna Dondi ◽

Carlotta Biagi ◽

...

Keyword(s):

Escherichia Coli ◽

Metropolitan Area ◽

Bacterial Infections ◽

Asymptomatic Bacteriuria ◽

Multidrug Resistant ◽

Resistance Rate ◽

First Line ◽

E Coli ◽

First Line Therapy ◽

Tract Infections

Urinary tract infections (UTIs) are among the most common bacterial infections in children, and Escherichia coli is the main pathogen responsible. Several guidelines, including the recently updated Italian guidelines, recommend amoxicillin-clavulanic acid (AMC) as a first-line antibiotic therapy in children with febrile UTIs. Given the current increasing rates of antibiotic resistance worldwide, this study aimed to investigate the three-year trend in the resistance rate of E. coli isolated from pediatric urine cultures (UCs) in a metropolitan area of northern Italy. We conducted a retrospective review of E. coli-positive, non-repetitive UCs collected in children aged from 1 month to 14 years, regardless of a diagnosis of UTI, catheter colonization, urine contamination, or asymptomatic bacteriuria. During the study period, the rate of resistance to AMC significantly increased from 17.6% to 40.2% (p < 0.001). Ciprofloxacin doubled its resistance rate from 9.1% to 16.3% (p = 0.007). The prevalence of multidrug-resistant E. coli rose from 3.9% to 9.2% (p = 0.015). The rate of resistance to other considered antibiotics remained stable, as did the prevalence of extended spectrum beta-lactamases and extensively resistant E. coli among isolates. These findings call into question the use of AMC as a first-line therapy for pediatric UTIs in our population, despite the indications of recent Italian guidelines.

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Impact of On-Farm Interventions against CTX-Resistant Escherichia coli on the Contamination of Carcasses before and during an Experimental Slaughter

Antibiotics ◽

10.3390/antibiotics10030228 ◽

2021 ◽

Vol 10 (3) ◽

pp. 228

Author(s):

Michaela Projahn ◽

Jana Sachsenroeder ◽

Guido Correia-Carreira ◽

Evelyne Becker ◽

Annett Martin ◽

...

Keyword(s):

Escherichia Coli ◽

Broiler Chickens ◽

Stocking Density ◽

Resistant Bacteria ◽

Control Group ◽

Cross Contamination ◽

High Prevalence ◽

On Farm ◽

The Impact ◽

Slow Growing

Cefotaxime (CTX)-resistant Enterobacteriaceae are still an ongoing challenge in human and veterinary health. High prevalence of these resistant bacteria is detected in broiler chickens and the prevention of their dissemination along the production pyramid is of major concern. The impact of certain on-farm interventions on the external bacterial contamination of broiler chickens, as well as their influence on single processing steps and (cross-) contamination, have not yet been evaluated. Therefore, we investigated breast skin swab samples of broiler chickens before and during slaughter at an experimental slaughter facility. Broiler chickens were previously challenged with CTX-resistant Escherichia coli strains in a seeder-bird model and subjected to none (control group (CG)) or four different on-farm interventions: drinking water supplementation based on organic acids (DW), slow growing breed Rowan × Ranger (RR), reduced stocking density (25 kg/sqm) and competitive exclusion with Enterobacteriales strain IHIT36098(CE). Chickens of RR, 25 kg/sqm, and CE showed significant reductions of the external contamination compared to CG. The evaluation of a visual scoring system indicated that wet and dirty broiler chickens are more likely a vehicle for the dissemination of CTX-resistant and total Enterobacteriaceae into the slaughterhouses and contribute to higher rates of (cross-) contamination during processing.

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Mechanistic insights into synergy between nalidixic acid and tetracycline against clinical isolates of Acinetobacter baumannii and Escherichia coli

Communications Biology ◽

10.1038/s42003-021-02074-5 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Amit Gaurav ◽

Varsha Gupta ◽

Sandeep K. Shrivastava ◽

Ranjana Pathania

Keyword(s):

Escherichia Coli ◽

Acinetobacter Baumannii ◽

Nalidixic Acid ◽

Bacterial Infections ◽

Clinical Isolates ◽

Hospital Environment ◽

Infection Model ◽

Drug Resistant ◽

E Coli

AbstractThe increasing prevalence of antimicrobial resistance has become a global health problem. Acinetobacter baumannii is an important nosocomial pathogen due to its capacity to persist in the hospital environment. It has a high mortality rate and few treatment options. Antibiotic combinations can help to fight multi-drug resistant (MDR) bacterial infections, but they are rarely used in the clinics and mostly unexplored. The interaction between bacteriostatic and bactericidal antibiotics are mostly reported as antagonism based on the results obtained in the susceptible model laboratory strain Escherichia coli. However, in the present study, we report a synergistic interaction between nalidixic acid and tetracycline against clinical multi-drug resistant A. baumannii and E. coli. Here we provide mechanistic insight into this dichotomy. The synergistic combination was studied by checkerboard assay and time-kill curve analysis. We also elucidate the mechanism behind this synergy using several techniques such as fluorescence spectroscopy, flow cytometry, fluorescence microscopy, morphometric analysis, and real-time polymerase chain reaction. Nalidixic acid and tetracycline combination displayed synergy against most of the MDR clinical isolates of A. baumannii and E. coli but not against susceptible isolates. Finally, we demonstrate that this combination is also effective in vivo in an A. baumannii/Caenorhabditis elegans infection model (p < 0.001)

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Impact of Feed Supplementation with Antimicrobial Agents on Growth Performance of Broiler Chickens, Clostridium perfringens and Enterococcus Counts, and Antibiotic Resistance Phenotypes and Distribution of Antimicrobial Resistance Determinants in Escherichia coli Isolates

Applied and Environmental Microbiology ◽

10.1128/aem.01086-07 ◽

2007 ◽

Vol 73 (20) ◽

pp. 6566-6576 ◽

Cited By ~ 103

Author(s):

Moussa S. Diarra ◽

Fred G. Silversides ◽

Fatoumata Diarrassouba ◽

Jane Pritchard ◽

Luke Masson ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Broiler Chickens ◽

Antimicrobial Agents ◽

Growth Promoters ◽

Feed Supplementation ◽

E Coli ◽

Resistance Determinants ◽

Class 1

ABSTRACT The effects of feed supplementation with the approved antimicrobial agents bambermycin, penicillin, salinomycin, and bacitracin or a combination of salinomycin plus bacitracin were evaluated for the incidence and distribution of antibiotic resistance in 197 commensal Escherichia coli isolates from broiler chickens over 35 days. All isolates showed some degree of multiple antibiotic resistance. Resistance to tetracycline (68.5%), amoxicillin (61.4%), ceftiofur (51.3%), spectinomycin (47.2%), and sulfonamides (42%) was most frequent. The levels of resistance to streptomycin, chloramphenicol, and gentamicin were 33.5, 35.5, and 25.3%, respectively. The overall resistance levels decreased from day 7 to day 35 (P < 0.001). Comparing treatments, the levels of resistance to ceftiofur, spectinomycin, and gentamicin (except for resistance to bacitracin treatment) were significantly higher in isolates from chickens receiving feed supplemented with salinomycin than from the other feeds (P < 0.001). Using a DNA microarray analysis capable of detecting commonly found antimicrobial resistance genes, we characterized 104 tetracycline-resistant E. coli isolates from 7- to 28-day-old chickens fed different growth promoters. Results showed a decrease in the incidence of isolates harboring tet(B), bla TEM, sulI, and aadA and class 1 integron from days 7 to 35 (P < 0.01). Of the 84 tetracycline-ceftiofur-resistant E. coli isolates, 76 (90.5%) were positive for bla CMY-2. The proportions of isolates positive for sulI, aadA, and integron class 1 were significantly higher in salinomycin-treated chickens than in the control or other treatment groups (P < 0.05). These data demonstrate that multiantibiotic-resistant E. coli isolates can be found in broiler chickens regardless of the antimicrobial growth promoters used. However, the phenotype and the distribution of resistance determinants in E. coli can be modulated by feed supplementation with some of the antimicrobial agents used in broiler chicken production.

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Alteration of the phagocytosis and antimicrobial defense of Octodonta nipae (Coleoptera: Chrysomelidae) pupae to Escherichia coli following parasitism by Tetrastichus brontispae (Hymenoptera: Eulophidae)

Bulletin of Entomological Research ◽

10.1017/s0007485318000780 ◽

2018 ◽

Vol 109 (2) ◽

pp. 248-256

Author(s):

E. Meng ◽

J. Li ◽

B. Tang ◽

Y. Hu ◽

T. Qiao ◽

...

Keyword(s):

Escherichia Coli ◽

Immune Response ◽

Immune Responses ◽

Mrna Expression ◽

Bactericidal Activity ◽

Bacterial Infections ◽

Expression Level ◽

Primary Immune Response ◽

Mrna Expression Level ◽

E Coli

AbstractAlthough parasites and microbial pathogens are both detrimental to insects, little information is currently available on the mechanism involved in how parasitized hosts balance their immune responses to defend against microbial infections. We addressed this in the present study by comparing the immune response between unparasitized and parasitized pupae of the chrysomelid beetle, Octodonta nipae (Maulik), to Escherichia coli invasion. In an in vivo survival assay, a markedly reduced number of E. coli colony-forming units per microliter was detected in parasitized pupae at 12 and 24 h post-parasitism, together with decreased phagocytosis and enhanced bactericidal activity at 12 h post-parasitism. The effects that parasitism had on the mRNA expression level of selected antimicrobial peptides (AMPs) of O. nipae pupae showed that nearly all transcripts of AMPs examined were highly upregulated during the early and late parasitism stages except defensin 2B, whose mRNA expression level was downregulated at 24 h post-parasitism. Further elucidation on the main maternal fluids responsible for alteration of the primary immune response against E. coli showed that ovarian fluid increased phagocytosis at 48 h post-injection. These results indicated that the enhanced degradation of E. coli in parasitized pupae resulted mainly from the elevated bactericidal activity without observing the increased transcripts of target AMPs. This study contributes to a better understanding of the mechanisms involved in the immune responses of a parasitized host to bacterial infections.

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Antimicrobial Resistance of Escherichia coli and Salmonella isolated from Raw Retail Broiler Chickens in Zambia

10.21203/rs.2.14062/v1 ◽

2019 ◽

Author(s):

Elizabeth Muligisa Muonga ◽

Geoffrey Mainda ◽

Mercy Mukuma ◽

Geoffrey Kwenda ◽

Bernard Hang'ombe ◽

...

Keyword(s):

Public Health ◽

Escherichia Coli ◽

Antimicrobial Resistance ◽

Broiler Chickens ◽

Target Genes ◽

Health Concern ◽

Poultry Production ◽

Public Health Concern ◽

E Coli ◽

Open Markets

Abstract Background Antimicrobial resistance (AMR) of foodborne pathogens is of public health concern, especially in developing countries like Zambia. This study was undertaken to determine the resistance profiles of Escherichia coli ( E. coli ) and Salmonella isolated from dressed broiler chickens purchased from open markets and supermarkets in Zambia.Results A total of 189 E. coli and five Salmonella isolates were isolated. Identification and confirmation of the isolates was done using Analytical Profile Index (API 20E) (Biomerieux ® ) and 16S rRNA sequencing. Antimicrobial susceptibility tests (AST) were performed using the Kirby Bauer disk diffusion technique using a panel of 10 different antibiotics and multiplex PCR was used to determine the presence of three target genes encoding for resistance: tetA, Sul1 and CTXM. AST results were entered and analyzed in WHONET 2018 software. A total of 189 E. coli and five Salmonella isolates were identified. Among the E. coli isolates, Tetracycline recorded the highest resistance of 79.4%, followed by Ampicillin 51.9%, Trimethoprim/Sulfamethoxazole 49.7%, Nalidixic Acid 24.3%, Chloramphenicol 16.4%, Cefotaxime 16.4%, Ciprofloxacin 10.1%, Colistin 7.4%, Amoxicillin/Clavulanic acid 6.9%, and Imipenem 1.1%. Two of the five Salmonella isolates were resistant to at least one antibiotic. Forty- seven (45.2%) of the isolates possessed at least one of the targeted resistance genes.Conclusion This study has demonstrated the presence of AMR E. coli and Salmonella on raw broiler chickens from both open markets and supermarkets. Such resistance is of public health concern and measures need to be put in place to regulate the use of these antimicrobials in poultry production.

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