scholarly journals Gut carriage of antimicrobial resistance genes in women exposed to small-scale poultry farms in rural Uganda: a feasibility study

2020 ◽  
Author(s):  
Ana A. Weil ◽  
Meti D. Debela ◽  
Daniel M. Muyanja ◽  
Bernard Kakuhikire ◽  
Charles Baguma ◽  
...  

ABSTRACTBackgroundAntibiotic use as growth promoters for livestock is presumed to be a major contributor to the acquisition of antimicrobial resistance (AMR) genes in humans, yet data evaluating AMR patterns in the setting of animal exposure are limited to observational studies that do not capture data from prior to livestock introduction.MethodsWe performed a feasibility study by recruiting a subset of women in a delayed-start randomized controlled trial of small-scale chicken farming in order to examine the prevalence of clinically-relevant AMR genes. Stool samples were obtained at baseline and one year from five intervention women who received chickens at the start of the study, six control women who did not receive chickens until the end of the study, and from chickens provided to the control group at the end of the study. Stool was screened for 87 clinically significant AMR genes using a commercially available qPCR array (Qiagen).ResultsChickens harbored 23 AMR genes from classes also found in humans as well as vancomycin and additional β-lactamase resistance genes. After one year of exposure to chickens, six new AMR genes were detected in controls and seven new AMR genes were detected in the intervention group. Women who had direct contact with the chickens sampled in the study had greater similarities in AMR resistance gene patterns to chickens than those who did not have direct contact with chickens sampled (p = 0.006). There was a trend towards increased similarity in AMR gene patterns with chickens at one year (p = 0.12).ConclusionsChickens and humans in this study harbored AMR genes from many antimicrobial classes at both baseline and follow up timepoints. Studies designed to evaluate human AMR genes in the setting of animal exposure should account for high baseline AMR rates, and consider collecting concomitant animal samples, human samples, and environmental samples over time to determine the directionality and source of AMR genes.Trial registration: ClinicalTrials.gov Identifier: NCT02619227

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S366-S366
Author(s):  
Meti D Debela ◽  
Daniel M Muyanja ◽  
Bernard Kakuhikire ◽  
Charles Baguma ◽  
David R Bangsberg ◽  
...  

Abstract Background Antibiotic use in livestock farming is thought to be a major contributor to the spread of antimicrobial resistance (AMR) genes in humans. However, quantitative data in this in this field are rare. To address this gap in the literature, we examined the prevalence of clinically important AMR genes before and after the introduction of chicken farming among women in rural Uganda. Methods We recruited a subset of women participating in a waitlist-randomized controlled trial of small-scale hybrid chicken farming in rural Uganda. Tetracycline is routinely administered to chicks during brooding. Stool samples before and one year after chicken introduction were obtained from six women randomized to the control arm, from five women randomized to the intervention arm, and from chickens. Microbial DNA was extracted from chicken and human stool and screened for 87 AMR genes using validated qPCR arrays (Qiagen). Results The median age was 35 years. At baseline, 10 of the women reported animal contact, most commonly goats (n = 8), free ranging village chickens (n = 7), cats (n = 4), and dogs (n = 4). During baseline testing of the women’s stool, we detected 18 genes conferring AMR to aminoglycosides, fluoroquinolones, macrolides, lincosamides, streptogramin B, Class A-C β-lactamases and tetracycline efflux pumps. Chickens harbored 23 AMR genes from the same classes as found in humans, and were also found to have vancomycin resistance genes (Van B and C) and Group D β-lactamases (OXA-58 and OXA-10). At one year, six new AMR genes emerged in controls, including one present in chickens; CTX-M-1, a Class A β-lactamase. In contrast, seven new AMR genes emerged in the intervention group, including four present in chickens: SHV, SHV(238G240E), (Class A β lactamases) and QnrS, QnrB-5 (fluoroquinolone resistance genes). Two AMR genes gained by both control and intervention groups were not present in chickens. Conclusion Women exposed to small-scale chicken farming acquired more AMR genes compared with unexposed participants. Chickens harbored many of the genes that emerged in humans. Introduction of antibiotic-treated animals may result in the transfer of AMR genes from animals to humans, even among humans exposed to a wide range of animals at baseline. Disclosures All authors: No reported disclosures.


PLoS ONE ◽  
2020 ◽  
Vol 15 (6) ◽  
pp. e0229699
Author(s):  
Ana A. Weil ◽  
Meti D. Debela ◽  
Daniel M. Muyanja ◽  
Bernard Kakuhikire ◽  
Charles Baguma ◽  
...  

PLoS ONE ◽  
2021 ◽  
Vol 16 (5) ◽  
pp. e0252261
Author(s):  
Ana A. Weil ◽  
Meti D. Debela ◽  
Daniel M. Muyanja ◽  
Bernard Kakuhikire ◽  
Charles Baguma ◽  
...  

2021 ◽  
Author(s):  
Jenna M Swarthout ◽  
Erica R Fuhrmeister ◽  
Latifah Hamzah ◽  
Angela Harris ◽  
Mir A. Ahmed ◽  
...  

Background Low- and middle-income countries (LMICs) bear the largest mortality burden due to antimicrobial-resistant infections. Small-scale animal production and free-roaming domestic animals are common in many LMICs, yet data on zoonotic exchange of gut bacteria and antimicrobial resistance genes (ARGs) in low-income communities are sparse. Differences between rural and urban communities in population density, antibiotic use, and cohabitation with animals likely influence the frequency of transmission of gut bacterial communities and ARGs between humans and animals. Here, we determined the similarity in gut microbiomes, using 16S rRNA gene amplicon sequencing, and resistomes, using long-read metagenomics, between humans, chickens, and goats in rural compared to urban Bangladesh. Results Gut microbiomes were more similar between humans and chickens in rural (where cohabitation is more common) compared to urban areas, but there was no difference for humans and goats. Urbanicity did not impact the similarity of human and animal resistomes; however, ARG abundance was higher in urban animals compared to rural animals. We identified substantial overlap of ARG alleles in humans and animals in both settings. Humans and chickens had more overlapping ARG alleles than humans and goats. All fecal hosts carried ARGs on contigs classified as potentially pathogenic bacteria, including Escherichia coli, Campylobacter jejuni, Clostridiodes difficile, and Klebsiella pneumoniae. Conclusions While the development of antimicrobial resistance in animal gut microbiomes and subsequent transmission to humans has been demonstrated in intensive farming environments and high-income countries, evidence of zoonotic exchange of antimicrobial resistance in LMIC communities is lacking. This research provides genomic evidence of overlap of antimicrobial resistance genes between humans and animals, especially in urban communities, and highlights chickens as important reservoirs of antimicrobial resistance. Chicken and human gut microbiomes were more similar in rural Bangladesh, where cohabitation is more common. Incorporation of long-read metagenomics enabled characterization of bacterial hosts of resistance genes, which has not been possible in previous culture-independent studies using only short-read sequencing. These findings highlight the importance of developing strategies for combatting antimicrobial resistance that account for chickens being reservoirs of ARGs in community environments, especially in urban areas.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
R. V. Pereira ◽  
C. Foditsch ◽  
J. D. Siler ◽  
S. C. Dulièpre ◽  
C. Altier ◽  
...  

Abstract The objective of this study was to evaluate the longitudinal effect of enrofloxacin or tulathromycin use in calves at high risk of bovine respiratory disease (BRD) on antimicrobial resistance genes and mutation in quinolone resistance-determining regions (QRDR) in fecal E. coli. Calves at high risk of developing BRD were randomly enrolled in one of three groups receiving: (1) enrofloxacin (ENR; n = 22); (2) tulathromycin (TUL; n = 24); or (3) no treatment (CTL; n = 21). Fecal samples were collected at enrollment and at 7, 28, and 56 days after beginning treatment, cultured for Escherichiacoli (EC) and DNA extracted. Isolates were screened for cephalosporin, quinolone and tetracycline resistance genes using PCR. QRDR screening was conducted using Sanger sequencing. The only resistance genes detected were aac(6′)Ib-cr (n = 13), bla-CTX-M (n = 51), bla-TEM (n = 117), tetA (n = 142) and tetB (n = 101). A significantly higher detection of gyrA mutated at position 248 at time points 7 (OR = 11.5; P value = 0.03) and 28 (OR = 9.0; P value = 0.05) was observed in the ENR group when compared to calves in the control group. Our findings support a better understanding of the potential impacts from the use of enrofloxacin in calves on the selection and persistence of resistance.


PLoS ONE ◽  
2020 ◽  
Vol 15 (6) ◽  
pp. e0234127 ◽  
Author(s):  
Ana Paula Christoff ◽  
Aline Fernanda Rodrigues Sereia ◽  
Giuliano Netto Flores Cruz ◽  
Daniela Carolina de Bastiani ◽  
Vanessa Leitner Silva ◽  
...  

Microbiology ◽  
2020 ◽  
Vol 166 (8) ◽  
pp. 727-734 ◽  
Author(s):  
Ana Luisa Andrade-Oliveira ◽  
Ciro César Rossi ◽  
Thaysa Souza-Silva ◽  
Marcia Giambiagi-deMarval

Staphylococcus nepalensis is a commensal bacterium from the oral microbiota of domestic cats, with a still obscure clinical importance. In this work, we analysed the ability of feline strains of S. nepalensis to transfer antimicrobial resistance genes to Staphylococcus aureus isolated from humans through plasmids. To this end, we first analysed all publicly available genomes from cat staphylococci using computational methods to build a pan-resistome. Genes that encode resistance to erythromycin, gentamicin, mupirocin and tetracycline, common to human and cat staphylococci and previously described to be located in mobile genetic elements, were chosen for the next analyses. We studied 15 strains of S. nepalensis , which were shown to be genetically different by GTG5-PCR. As observed by disc diffusion, resistance to tetracycline was widespread (80 %), followed by resistance to erythromycin (40 %), gentamicin (27 %) and mupirocin (7 %). The strains were positive for several antimicrobial resistance genes and more than half of them harboured plasmids. The loss of plasmids and resistance genes in some strains were induced by stress with SDS. Through conjugation experiments, we observed that these plasmids can be transferred to S. aureus , thus increasing its potential to resist drug therapy. Our findings show that S. nepalensis , an underestimated inhabitant of the cat microbiota, can be a reservoir of antimicrobial resistance genes for S. aureus and, like many other staphylococci, be an overlooked and silent threat to their animal hosts and humans living with them.


2021 ◽  
Vol 70 (11) ◽  
Author(s):  
Lii-Tzu Wu ◽  
Xin-Xia Wu ◽  
Se-Chin Ke ◽  
Yi-Pei Lin ◽  
Ying-Chen Wu ◽  
...  

Introduction. Antimicrobial resistance associated with animal hosts is easily transmitted to humans either by direct contact with resistant organisms or by transferring resistance genes into human pathogens. Gap statement. There are limited studies on antimicrobial resistance genes and genetic elements of multidrug-resistant (MDR) Escherichia coli in veterinary hospitals in Taiwan. Aim. The aim of this study was to investigate antimicrobial resistance genes in multidrug-resistant Escherichia coli from animals. Methodology. Between January 2014 and August 2015, 95 multidrug-resistant Escherichia coli isolates were obtained from pigs (n=66), avians (n=18), and other animals (n=11) in a veterinary hospital in Taiwan. Susceptibility testing to 24 antimicrobial agents of 14 antimicrobial classes was performed. Antimicrobial resistance genes, integrons, and insertion sequences were analysed by polymerase chain reaction and nucleotide sequencing. Pulsed-field gel electrophoresis (PFGE), and multi-locus sequence typing were used to explore the clonal relatedness of the study isolates. Results. Different antimicrobial resistance genes found in these isolates were associated with resistance to β-lactams, tetracycline, phenicols, sulfonamides, and aminoglycosides. Fifty-five of 95 E. coli isolates (55/95, 57.9 %) were not susceptible to extended-spectrum cephalosporins, and bla CTX-M-55 (11/55, 20.0 %) and bla CMY-2 (40/55, 72.7 %) were the most common extended-spectrum β-lactamase (ESBL) and AmpC genes, respectively. Both bla CTX-M and bla CMY-2 were present on conjugative plasmids that contained the insertion sequence ISEcp1 upstream of the bla genes. Plasmid-mediated FOX-3 β-lactamase-producing E. coli was first identified in Taiwan. Forty isolates (40/95, 42 %) with class 1 integrons showed seven resistance phenotypes. Genotyping of 95 E. coli isolates revealed 91 different XbaI pulsotypes and 52 different sequence types. PFGE analysis revealed no clonal outbreaks in our study isolates. Conclusion. This study showed a high diversity of antimicrobial resistance genes and genotypes among MDR E. coli isolated from diseased livestock in Taiwan. To our knowledge, this is the first report of plasmid-mediated ESBL in FOX-3 β-lactamase-producing E. coli isolates in Taiwan. MDR E. coli isolates from animal origins may contaminate the environment, resulting in public health concerns, indicating that MDR isolates from animals need to be continuously investigated.


2021 ◽  
Vol 70 (7) ◽  
Author(s):  
Suleepon Poomchuchit ◽  
Anusak Kerdsin ◽  
Peechanika Chopjitt ◽  
Parichart Boueroy ◽  
Rujirat Hatrongjit ◽  
...  

Introduction. The emergence and spread of non-typhoidal Salmonella enterica (NTS) serovars resistant to fluoroquinolones and third- and higher-generation cephalosporins is a matter of great concern. Antimicrobial-resistant NTS is increasingly being discovered in humans, animals, food animals, food products, and agricultural environments. Pigs are considered a major reservoir of antimicrobial-resistant Salmonella spp. Hypothesis/Gap Statement. Fluoroquinolone-resistant Salmonella spp. warrant further surveillance and characterization for a better understanding of the bacteria isolated from animals. Aim. NTS isolated from pork from slaughterhouses across Thailand were characterized in terms of their serovars; resistance to fluoroquinolones, third-generation cephalosporins, and carbapenems; and antimicrobial resistance genes. Methodology. A total of 387 NTS isolates, collected from slaughtered pigs in ten provinces across Thailand between 2014 and 2015, were characterized based on their serovars, antimicrobial resistance genes, and susceptibility to fluoroquinolones, third-generation cephalosporins, and carbapenems. Results. Among all NTS isolates, S. enterica serovar Rissen was predominant. Antimicrobial resistance was exhibited in 93/387 isolates (24 %). Although 24 (6.2 %) isolates were susceptible to all the tested antimicrobials, they were found to possess β-lactamase genes, such as bla TEM, bla SHV, or bla CTX-M. Mobilized colistin-resistant genes (mcr) and resistance to colistin were not observed in any tested isolate. Carbapenem resistance was detected in ten isolates (10.7 %); however, bla KPC, bla NDM, bla OXA-48-like, and bla IMP were not present. Among the 93 antimicrobial-resistant isolates, 87.1 % showed fluoroquinolone resistance with the quinolone resistance gene (qnrS) combined with topoisomerase genes parC (T57S) or gyrA (S83E/Y and D124E/G) substitutions, or topoisomerase gene substitutions alone. Conclusion. We found high fluoroquinolone resistance rates among the NTS isolates from pigs from slaughterhouses. The fluoroquinolone resistance mechanism in NTS was associated with the combination of qnrS and substitutions in gyrA, parC, or both. To prevent the transmission of antimicrobial-resistant NTS between animals and humans, continuous monitoring, surveillance, and regulation of Salmonella in the pork supply chain are pivotal.


2021 ◽  
Vol 9 (4) ◽  
pp. 707
Author(s):  
J. Christopher Noone ◽  
Fabienne Antunes Ferreira ◽  
Hege Vangstein Aamot

Our culture-independent nanopore shotgun metagenomic sequencing protocol on biopsies has the potential for same-day diagnostics of orthopaedic implant-associated infections (OIAI). As OIAI are frequently caused by Staphylococcus aureus, we included S. aureus genotyping and virulence gene detection to exploit the protocol to its fullest. The aim was to evaluate S. aureus genotyping, virulence and antimicrobial resistance genes detection using the shotgun metagenomic sequencing protocol. This proof of concept study included six patients with S. aureus-associated OIAI at Akershus University Hospital, Norway. Five tissue biopsies from each patient were divided in two: (1) conventional microbiological diagnostics and genotyping, and whole genome sequencing (WGS) of S. aureus isolates; (2) shotgun metagenomic sequencing of DNA from the biopsies. Consensus sequences were analysed using spaTyper, MLST, VirulenceFinder, and ResFinder from the Center for Genomic Epidemiology (CGE). MLST was also compared using krocus. All spa-types, one CGE and four krocus MLST results matched Sanger sequencing results. Virulence gene detection matched between WGS and shotgun metagenomic sequencing. ResFinder results corresponded to resistance phenotype. S. aureus spa-typing, and identification of virulence and antimicrobial resistance genes are possible using our shotgun metagenomics protocol. MLST requires further optimization. The protocol has potential application to other species and infection types.


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