Low Occurrence of Extended‐Spectrum β‐lactamase‐Producing Escherichia coli in Finnish Food‐Producing Animals

ESBL/AmpC‐producing Escherichia coli is increasingly isolated from humans and animals worldwide. The occurrence of ESBL/AmpC‐producing E. coli was studied in food‐producing animals in Finland, a country with a low and controlled use of antimicrobials in meat production chain. A total of 648 cattle, 531 pig, 495 broiler and 35 turkey faecal samples were collected from four Finnish slaughterhouses to determine the presence of extended‐spectrum β‐lactamase (ESBL/AmpC)‐producing E. coli. In addition, 260 broiler and 15 turkey samples were screened for carbapenemase‐producing E. coli. Susceptibility to different class of cephalosporins and meropenem was determined with disc diffusion tests according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Determination of ESBL/AmpC production was performed with a combination disc diffusion test according to the recommendations of the European Food Safety Authority (EFSA). Plasmidic bla_ESBL/AmpC genes were characterized by polymerase chain reaction and sequencing. A collection of isolates producing AmpC enzyme but not carrying plasmidic bla_AmpC was analysed by PCR and sequencing for possible chromosomal ampC promoter area mutations. Altogether ESBL/AmpC‐producing E. coli was recovered from five cattle (0.8%), eight pig (1.5%) and 40 broiler samples (8.1%). No ESBL/AmpC‐producing E. coli was found in turkey samples. Carbapenem resistance was not detected. Altogether ESBL/AmpC‐producing E. coli was found on 4 (2.0%), 3 (4.5%) and 14 (25%) cattle, pig and broiler farms, respectively. From cattle samples 3 (27%) bla_CTX‐M‐1 and from broiler samples 13 (33%) bla_CTX‐M‐1 and 22 (55%) bla_CMY‐2 gene‐carrying isolates were detected. In pigs, no plasmidic bla_ESBL/AmpC gene‐carrying isolates were found. In all analysed isolates, the same mutations in the promoter region of chromosomal ampC were detected. The results showed low occurrence of ESBL/AmpC‐producing E. coli in Finnish food‐producing animals. In pigs, plasmidic bla_ESBL/AmpC‐carrying E. coli was not detected at all.     

Prevalence of AmpC‐ and Extended‐Spectrum β‐Lactamase‐Harbouring Enterobacteriaceae in Faecal Flora of a Healthy Domestic Canine Population

In order to estimate the prevalence of AmpC‐ and ESBL β‐lactamase‐producing Enterobacteriaceae in the faecal flora of a healthy domestic canine population, faecal samples were obtained from healthy dogs receiving routine parasitology screening at the Ohio State University Veterinary Medical Center, between January 2013 and April 2013. Samples were screened for the presence of AmpC and ESBL β‐lactamase phenotypes, and the clinically important genotypes, bla_CMY and bla_CTX‐M, were confirmed via conventional PCR. Minimum inhibitory concentrations were determined for isolates and plasmids were characterized. Two hundred and twelve canine faecal samples were screened, of which 30 harboured isolates carrying the AmpC bla_CMY, representing 14.2% of the population (95% CI: 9.4–18.9%). Nine samples harboured isolates that carried the ESBL bla_CTX‐M, representing 4.2% of the population (95% CI: 1.5–7.0%). Isolates containing bla_CMY harboured multiple plasmid replicon types, while isolates containing bla_CTX‐M harboured few plasmid replicon types. Our results suggest that domestic dogs may serve as a reservoir for extended‐spectrum cephalosporin resistance genes for other domestic animal populations as well as for their human companions. This represents a potential veterinary and public health risk that warrants further investigation and continued surveillance to ascertain the nature and extent of the risk. The high level of diversity of plasmid content among isolates harbouring bla_CMY suggests broader dissemination relative to bla_CTX‐M isolates.     

Extended‐spectrum beta‐lactamase‐producing Escherichia coli in common vampire bats Desmodus rotundus and livestock in Peru

Antibiotic resistance mediated by bacterial production of extended‐spectrum beta‐lactamase (ESBL) is a global threat to public health. ESBL resistance is most commonly hospital‐acquired; however, infections acquired outside of hospital settings have raised concerns over the role of livestock and wildlife in the zoonotic spread of ESBL‐producing bacteria. Only limited data are available on the circulation of ESBL‐producing bacteria in animals. Here, we report ESBL‐producing Escherichia coli in wild common vampire bats Desmodus rotundus and livestock near Lima, Peru. Molecular analyses revealed that most of this resistance resulted from the expression of bla_CTX‐M‐15 genes carried by plasmids, which are disseminating worldwide in hospital settings and have also been observed in healthy children of Peru. Multilocus sequence typing showed a diverse pool of E. coli strains carrying this resistance that were not always host species‐specific, suggesting sharing of strains between species or infection from a common source. This study shows widespread ESBL resistance in wild and domestic animals, supporting animal communities as a potential source of resistance. Future work is needed to elucidate the role of bats in the dissemination of antibiotic‐resistant strains of public health importance and to understand the origin of the observed resistance.     

Diversity of Plasmids and Antimicrobial Resistance Genes in Multidrug‐Resistant Escherichia coli Isolated from Healthy Companion Animals

The presence and transfer of antimicrobial resistance genes from commensal bacteria in companion animals to more pathogenic bacteria may contribute to dissemination of antimicrobial resistance. The purpose of this study was to determine antimicrobial resistance gene content and the presence of genetic elements in antimicrobial resistant Escherichia coli from healthy companion animals. In our previous study, from May to August, 2007, healthy companion animals (155 dogs and 121 cats) from three veterinary clinics in the Athens, GA, USA area were sampled and multidrug‐resistant E. coli (n = 36; MDR, resistance to ≥2 antimicrobial classes) were obtained. Of the 25 different plasmid replicon types tested by PCR, at least one plasmid replicon type was detected in 94% (34/36) of the MDR E. coli; four isolates contained as many as five different plasmid replicons. Nine replicon types (FIA, FIB, FII, I2, A/C, U, P, I1 and HI2) were identified with FIB, FII, I2 as the most common pattern. The presence of class I integrons (intI) was detected in 61% (22/36) of the isolates with eight isolates containing aminoglycoside‐ and/or trimethoprim‐resistance genes in the variable cassette region of intI. Microarray analysis of a subset of the MDR E. coli (n = 9) identified the presence of genes conferring resistance to aminoglycosides (aac, aad, aph and strA/B), β‐lactams (ampC, cmy, tem and vim), chloramphenicol (cat), sulfonamides (sulI and sulII), tetracycline [tet(A), tet(B), tet(C), tet(D) and regulator, tetR] and trimethoprim (dfrA). Antimicrobial resistance to eight antimicrobials (ampicillin, cefoxitin, ceftiofur, amoxicillin/clavulanic acid, streptomycin, gentamicin, sulfisoxazole and trimethoprim‐sulfamethoxazole) and five plasmid replicons (FIA, FIB, FII, I1 and I2) were transferred via conjugation. The presence of antimicrobial resistance genes, intI and transferable plasmid replicons indicate that E. coli from companion animals may play an important role in the dissemination of antimicrobial resistance, particularly to human hosts during contact.     

A Cross‐Sectional Study Examining the Prevalence and Risk Factors for Anti‐Microbial‐Resistant Generic Escherichia coli in Domestic Dogs that Frequent Dog Parks in Three Cities in South‐Western Ontario,Canada

Anti‐microbial resistance can threaten health by limiting treatment options and increasing the risk of hospitalization and severity of infection. Companion animals can shed anti‐microbial‐resistant bacteria that may result in the exposure of other dogs and humans to anti‐microbial‐resistant genes. The prevalence of anti‐microbial‐resistant generic Escherichia coli in the faeces of dogs that visited dog parks in south‐western Ontario was examined and risk factors for shedding anti‐microbial‐resistant generic E. coli identified. From May to August 2009, canine faecal samples were collected at ten dog parks in three cities in south‐western Ontario, Canada. Owners completed a questionnaire related to pet characteristics and management factors including recent treatment with antibiotics. Faecal samples were collected from 251 dogs, and 189 surveys were completed. Generic E. coli was isolated from 237 of the faecal samples, and up to three isolates per sample were tested for anti‐microbial susceptibility. Eighty‐nine percent of isolates were pan‐susceptible; 82.3% of dogs shed isolates that were pan‐susceptible. Multiclass resistance was detected in 7.2% of the isolates from 10.1% of the dogs. Based on multilevel multivariable logistic regression, a risk factor for the shedding of generic E. coli resistant to ampicillin was attending dog day care. Risk factors for the shedding of E. coli resistant to at least one anti‐microbial included attending dog day care and being a large mixed breed dog, whereas consumption of commercial dry and home cooked diets was protective factor. In a multilevel multivariable model for the shedding of multiclass‐resistant E. coli, exposure to compost and being a large mixed breed dog were risk factors, while consumption of a commercial dry diet was a sparing factor. Pet dogs are a potential reservoir of anti‐microbial‐resistant generic E. coli; some dog characteristics and management factors are associated with the prevalence of anti‐microbial‐resistant generic E. coli in dogs.     

Low prevalence of zoonotic multidrug‐resistant bacteria in veterinarians in a country with prudent use of antimicrobials in animals

The occurrence of multidrug‐resistant zoonotic bacteria in animals has been increasing worldwide. Working in close contact with livestock increases the risk of carriage of these bacteria. We investigated the occurrence of extended‐spectrum beta‐lactamase (ESBL) and plasmidic AmpC beta‐lactamase producing Enterobacteriaceae (ESBL/pAmpC‐PE) and livestock‐associated methicillin‐resistant Staphylococcus aureus (LA‐MRSA) in Finnish veterinarians (n = 320). In addition to microbiological samples, background information was collected. Bacterial whole genome sequencing was performed to deduce sequence types (STs), spa types and resistance genes of the isolates. In total, 3.0% (9/297) of the veterinarians carried ESBL producing Escherichia coli, with one ESBL producing E. coli isolate producing also AmpC. Seven different STs, sequences of several different plasmid groups as well as several different bla_ESBL/pAmpC genes existed in different combinations. No carbapenemase or colistin resistance genes were detected. MRSA was detected in 0.3% (1/320) of the samples. The strain belonged to LA‐MRSA clonal complex (CC) 398 (ST398, spa type 011, lacking Panton‐Valentine leukocidin genes). In conclusion, this study shows low carriage of multidrug‐resistant zoonotic bacteria in Finnish veterinarians. However, finding LA‐MRSA for the first time in a sample from a veterinarian in a country with prudent use of animal antimicrobials and regarding the recent rise of LA‐MRSA on Finnish pig farms, a strong recommendation to protect people working in close contact with animals carrying LA‐MRSA CC398 is given. Further studies are needed to explain why the prevalence of LA‐MRSA in veterinarians is lower in Finland than in other European countries.     

Extended‐Spectrum β‐lactam Resistance in the Enteric Flora of Patients at a Tertiary Care Medical Centre

The dissemination of Enterobacteriaceae expressing resistance to extended‐spectrum cephalosporins, which are therapeutically used in both human and veterinary medicine, is of critical concern. The normal commensal flora of food animals may serve as an important reservoir for the zoonotic food‐borne transmission of Enterobacteriaceae harbouring β‐lactam resistance. We hypothesized that the predominant AmpC and ESBL genes reported in US livestock and fresh retail meat products, bla_CMY‐2 and bla_CTX‐M, would also be predominant in human enteric flora. We recovered enteric flora from a convenience sample of patients included in a large tertiary medical centre's Clostridium difficile surveillance programme to screen for and estimate the frequency of carriage of AmpC and ESBL resistance genes. In‐ and outpatient diarrhoeic submissions (n = 692) received for C. difficile testing at the medical centre's clinical diagnostic laboratory from July to December, 2013, were included. Aliquoted to a transport swab, each submission was inoculated to MacConkey broth with cefotaxime, incubated at 37°C and then inoculated to MacConkey agars supplemented with cefoxitin and cefepime to select for the AmpC and ESBL phenotypes, with bla_CMY and bla_CTX‐M genotypes confirmed by PCR and sequencing. From the 692 diarrhoeic submissions, our selective culture yielded 184 isolates (26.6%) with reduced susceptibility to cefotaxime. Of these, 46 (6.7%) samples harboured commensal isolates carrying the AmpC bla_CMY. Another 21 (3.0%) samples produced isolates harbouring the ESBL bla_CTX‐M: 19 carrying CTX‐M‐15 and 2 with CTX‐M‐27. Our results indicate that β‐lactam resistance genes likely acquired through zoonotic food‐borne transmission are present in the enteric flora of this hospital‐associated population at lower levels than reported in livestock and fresh food products.     

Antimicrobial Resistance in Wildlife: Implications for Public Health

The emergence and spread of antimicrobial‐resistant (AMR) bacteria in natural environments is a major concern with serious implications for human and animal health. The aim of this study was to determine the prevalence of AMR Escherichia coli (E. coli) in wild birds and mammalian species. Thirty faecal samples were collected from each of the following wildlife species: herring gulls (Larus argentatus), black‐headed gulls (Larus ridibundus), lesser black‐back gulls (Larus fuscus), hybrid deer species (Cervus elaphus x Cervus nippon) and twenty‐six from starlings (Sturnus vulgaris). A total of 115 E. coli isolates were isolated from 81 of 146 samples. Confirmed E. coli isolates were tested for their susceptibility to seven antimicrobial agents by disc diffusion. In total, 5.4% (8/146) of samples exhibited multidrug‐resistant phenotypes. The phylogenetic group and AMR‐encoding genes of all multidrug resistance isolates were determined by PCR. Tetracycline‐, ampicillin‐ and streptomycin‐resistant isolates were the most common resistant phenotypes. The following genes were identified in E. coli: bla_TEM, strA, tet(A) and tet(B). Plasmids were identified in all samples that exhibited multidrug‐resistant phenotypes. This study indicates that wild birds and mammals may function as important host reservoirs and potential vectors for the spread of resistant bacteria and genetic determinants of AMR.     

Faecal carriage of extended spectrum beta-lactamase (ESBL) and New Delhi metallo beta-lactamase(NDM) producing Escherichia coli between piglets and pig farmworkers

A cross-sectional study on five organized pig farms was conducted to assess the faecal carriage of ESBL and blaNDM carbapenemase-producing E. coli in piglets and pig farmworkers. Faecal samples from piglets (n = 155) and pig farmworkers (n = 21) were processed for isolation and characterization of E. coli. A total of 124 E. coli isolates from piglets and 21 E. coli isolates pig farmworkers were recovered and screening for ESBL production showed that 44.4 % (55/124) of the isolates from piglets and 42.9 % (9/21) of the isolates from farmworkers were ESBL positive. The ESBL positive isolates from piglets and farmworkers harbored blaCTX-M and also co-harbored other beta-lactams, sulphonamide, quinolone and tetracycline resistance genes. Diarrhoeic (50%, 49/98) and crossbred piglets (52.7%, 39/74) harbored a significantly higher number of ESBL producing isolates than non-diarrhoeic (23.1 %, 6/26) and purebred piglets (32%, 16/50) (p < 0.05). Piglets and pig farmworkers harbored nine and two carbapenem-resistant isolates, respectively. Interestingly, two isolates from piglets and one isolate from farmworkers harbored the blaNDM gene. The blaNDM positive E. coli isolated from piglets and farmworkers of the same farm revealed similar antibacterial resistance patterns, resistant genes, sequence (ST-167) and plasmid type (IncX3). In India, carbapenems are not used in food animal treatment, hence carbapenem resistant E. coli in piglets possibly originated from the human contact or common environment and is of public health importance.     

In vitro assessment of chloramphenicol and florfenicol as second‐line antimicrobial agents in dogs

The aim of this study was to evaluate the potential of chloramphenicol and florfenicol as second‐line antimicrobial agents for treatment of infections caused by methicillin‐resistant Staphyococcus pseudintermedius (MRSP) and extended‐spectrum β‐lactamase (ESBL)‐producing Escherichia coli in dogs, through a systematic in vitro assessment of the pharmacodynamic properties of the two drugs. Minimum inhibitory concentrations (MIC) and phenicol resistance genes were determined for 169 S. pseudintermedius and 167 E. coli isolates. Minimum bactericidal concentrations (MBC), time‐killing kinetics, and postantibiotic effect (PAE) of both agents against wild‐type isolates of each species were assessed. For S. pseudintermedius, the chloramphenicol MIC₉₀ was 32 μg/mL. No florfenicol resistance was detected in this species (MIC_90 = 4 μg/mL). The MIC₉₀ of both agents against E. coli was 8 μg/mL. Resistance genes found were cat_pC221 in S. pseudintermedius and catA1 and/or floR in E. coli. The phenicols displayed a time‐dependent, mainly, bacteriostatic effect on both species. Prolonged PAEs were observed for S. pseudintermedius, and no PAEs were detected for E. coli. More research into determination of PK/PD targets of efficacy is needed to further assess the clinical use of chloramphenicol and florfenicol as second‐line agents in dogs, optimize dosage regimens, and set up species‐specific clinical break points.     

High‐Level Ciprofloxacin‐Resistant Campylobacter jejuni Isolates Circulating in Humans and Animals in Incheon,Republic of Korea

Campylobacter jejuni is one of the major causative pathogens of outbreaks or sporadic cases of diarrhoeal diseases worldwide. In this study, we compared the phenotypic and genetic characteristics of C. jejuni isolates of human and food‐producing animal origins in Korea and examined the genetic relatedness between these two groups of isolates. Regardless of isolation source, all C. jejuni isolates harboured four virulence genes, cadF, cdtB, ciaB and racR, whereas the wlaN and virB11 genes were more frequently observed in human isolates. Antimicrobial susceptibility testing showed that the majority of C. jejuni isolates displayed high‐level resistance to fluoroquinolone (95.2%) or tetracycline (76.2%) antibiotics, and 12.4% of isolates exhibited multidrug resistance (more than three classes of antibiotics tested). Pulsed‐field gel electrophoresis (PFGE) of all Campylobacter isolates revealed 51 different SmaI‐PFGE patterns and six major clusters containing both human and animal isolates. These results indicate that genetically diverse strains of C. jejuni with antimicrobial drug‐resistance and virulence properties have prevailed in Incheon. Nevertheless, some particular populations continue to circulate within the community, providing the evidence for an epidemiological link of C. jejuni infections between humans and food‐producing animals. Therefore, the continued monitoring and surveillance of C. jejuni isolates of human and food‐producing animal origins are required for public health and food safety.     

Antibiotic resistant Escherichia coli in southeastern Australian pig herds and implications for surveillance

To investigate public health implications of antibiotics to control post‐weaning scours, we surveyed 22 commercial pig herds in southeastern Australia. Fifty faecal samples per herd were collected from pre‐ and post‐weaned piglets. Presumptive Escherichia coli isolates were confirmed by MALDI‐TOF MS. Isolates (n = 325) were screened for susceptibility to 19 veterinary antibiotics using MIC broth microdilution. All 325 E. coli isolates underwent further testing against 27 antibiotics used in human medicine and were screened for ETEC adhesin and enterotoxin genes (F4 (K88), F5 (K99), F6 (987P), F18, F41, STa, STb, Stx2e and LT) by multiplex PCR. Isolates identified as phenotypically resistant to third‐generation cephalosporin (3GC) and aminoglycoside antibiotics were screened by multiplex PCR/reverse line blot to detect common β‐lactam and aminoglycosides resistance genes, confirmed by sequencing. Twenty (6.1%) of the E. coli isolates were resistant to 3GC antibiotics and 24 (7.4%) to the aminoglycoside antibiotic gentamicin. Genetic analysis revealed six different extended spectrum β‐lactamase (ESBL) genes (blaCTX‐M‐1, ‐14, ‐15, ‐27, blaSHV‐12 and blaCMY‐2‐like genes), four of which have not been previously reported in Australian pigs. Critically, the prevalence of 3GC resistance was higher in non‐pathogenic (non‐ETEC) isolates and those from clinically normal (non‐diarrhoeal) samples. This highlights the importance of non‐ETECE. coli as reservoirs of antimicrobial resistance genes in piglet pens. Antimicrobial resistance surveillance in pig production focused on diagnostic specimens from clinically‐affected animals might be potentially misleading. We recommend that surveillance for emerging antimicrobial resistance such as to 3GC antibiotics should include clinically healthy pigs.     

Confirmation of a Campylobacteriosis Outbreak Associated with Chicken Liver Pâté Using PFGE and WGS

In May 2012, an outbreak of campylobacteriosis occurred in southern Sweden at a wedding reception affecting 44 persons. A total of 17 cases were notified (13 were culture positive for Campylobacter spp.). Epidemiological investigation suspected chicken liver pâté as the source of infection. The liver pâté had been deliberately undercooked, lightly fried to keep the right texture and mixed with spices. Campylobacter isolates from six cases as well as three Campylobacter isolates from chicken flocks previously raised by the producer delivering the liver were subtyped using pulsed‐field gel electrophoresis and whole‐genome sequencing. Indistinguishable PFGE profiles were identified among five human and one chicken C. jejuni isolates as well among the two C. coli isolates, one from a human case and one from a chicken. WGS supported the PFGE findings; the six C. jejuni isolates belonged to one cluster. All these six isolates were of MLST type ST 50 (ST‐CC 21). This study highlights the importance of a combination of strict biosecurity at the flock‐level as well as adequate cooking of chicken liver to prevent transmission of Campylobacter to humans.     

Preliminary insights into the occurrence of similar clones of extended‐spectrum beta‐lactamase‐producing bacteria in humans,animals and the environment in Tanzania: A systematic review and meta‐analysis between 2005 and 2016

The emergence and spread of extended‐spectrum beta‐lactamase producing Enterobacteriaceae (ESBL‐PE) are complex and of the public health concern across the globe. This review aimed at assessing the ESBL‐PE clones circulating in humans, animals and the environment to provide evidence‐based insights for combating ESBL‐PE using One Health approach. Systematic search from Medline/PubMed, Google Scholar and African Journals Online was carried out and retrieved nine eligible articles (of 131) based on phenotypic and genotypic detection of ESBL‐PE between 2005 and 2016 in Tanzania. Analysis was performed using STATA 11.0 software to delineate the prevalence of ESBL‐PE, phenotypic resistance profiles and clones circulating in the three interfaces. The overall prevalence of ESBL‐PE in the three interfaces was 22.6% (95% CI: 21.1–24.2) with the predominance of Escherichia coli (E. coli) strains (51.6%). The majority of ESBL‐PE were resistant to the commonly used antimicrobials such as trimethoprim–sulfamethoxazole and tetracycline/doxycycline, 38%–55% were resistant to ciprofloxacin and all were sensitive to meropenem/imipenem. ESBL‐PE infections were more associated with deaths compared to non‐ESBL‐PE infections. Strikingly, E. coli ST38, ST131 and ST2852 were found to intersect variably across the three interfaces. The predominant allele, bla_{CTX‐M‐15,} was found mostly in the conjugative IncF plasmids connoting transmission potential. The high prevalence of ESBL‐PE and shared clones across the three interfaces, including the global E. coli ST131 clone, indicates wide and inter‐compartmental spread that calls for One Health genomic‐driven studies to track the resistome flow.     

Investigations of the Distribution and Persistence of Salmonella and Ciprofloxacin‐Resistant Escherichia coli in Turkey Hatcheries in the UK

This study aimed at gaining information on the presence of Salmonella in UK turkey hatcheries and possible epidemiological links between breeding farms, hatcheries and finishing farms. The presence of ciprofloxacin‐resistant E. coli in hatchery samples, as well as in faecal samples from farms, and trends in occurrence of resistance were also investigated. Over a 2 year‐period, four British turkey hatcheries were visited and intensively sampled for the presence of Salmonella and ciprofloxacin‐resistant E. coli. In two hatcheries, a link could be demonstrated between the presence of certain Salmonella serovars in the hatcheries and on breeding and finishing farms. Within the hatcheries, serovars linked to breeding farms were found more frequently in the poult processing and dispatch areas, whereas serovars identified as ‘resident hatchery contaminants’ were predominantly found inside the hatcher cabinets. Ciprofloxacin‐resistant isolates of S. Senftenberg were identified in one hatchery, which coincided with enrofloxacin treatment of some of the breeding flocks. Ciprofloxacin‐resistant E. coli was found in two hatcheries, and the majority of these isolates showed multidrug resistance.     

Spread of extended-spectrum beta-lactamase producing Escherichia coli isolates in Swedish broilers mediated by an incI plasmid carrying blaCTX-M-1

Background

The already high and increasing occurrence of extended-spectrum beta-lactamases (ESBL) producing Escherichia coli in European broiler populations is of concern due to the fact that third and fourth generation cephalosporins are deemed critically important in human medicine. In Sweden 34% of the broilers carry ESBL/pAmpC producing E. coli in their gut, despite the absence of a known selection pressure such as antimicrobial usages. The aim of the current study was to characterise a selection of E. coli strains carrying the bla_CTX-M-1, to determine if the spread was due to a specific clone.

Findings

Ten isolates carrying bla_CTX-M-1 from Swedish broilers belonged to eight different multi-locus sequence types with three isolates belonging to ST155. The ST155 isolates were identical as assessed by PFGE. The bla_CTX-M-1 was in all isolates carried on a plasmid of replicon type incI, which also transferred resistance to tetracycline and sulfamethoxazole.

Conclusion

The occurrence of ESBL-producing E. coli in the Swedish broilers is not due to the emergence of a single clone, but rather the spread of a specific incI plasmid carrying bla_CTX-M-1.     

The occurrence of Salmonella,extended‐spectrum β‐lactamase producing Escherichia coli and carbapenem resistant non‐fermenting Gram‐negative bacteria in a backyard poultry flock environment

Increase in the number of small‐scale backyard poultry flocks in the USA has substantially increased human‐to‐live poultry contact, leading to increased public health risks of the transmission of multi‐drug resistant (MDR) zoonotic and food‐borne bacteria. The objective of this study was to detect the occurrence of Salmonella and MDR Gram‐negative bacteria (GNB) in the backyard poultry flock environment. A total of 34 backyard poultry flocks in Washington State (WA) were sampled. From each flock, one composite coop sample and three drag swabs from nest floor, waterer‐feeder, and a random site with visible faecal smearing, respectively, were collected. The samples were processed for isolation of Salmonella and other fermenting and non‐fermenting GNB under ceftiofur selection. Each isolate was identified to species level using MALDI‐TOFF and tested for resistance against 16 antibiotics belonging to eight antibiotic classes. Salmonella serovar 1,4,[5],12:i:‐ was isolated from one (3%) out of 34 flocks. Additionally, a total of 133 ceftiofur resistant (Cef^R) GNB including Escherichia coli (53), Acinetobacter spp. (45), Pseudomonas spp. (22), Achromobacter spp. (8), Bordetella trematum (1), Hafnia alvei (1), Ochrobactrum intermedium (1), Raoultella ornithinolytica (1), and Stenotrophomonas maltophilia (1) were isolated. Of these, 110 (82%) isolates displayed MDR. Each flock was found positive for the presence of one or more Cef^R GNB. Several MDR E. coli (n = 15) were identified as extended‐spectrum β‐lactamase (ESBL) positive. Carbapenem resistance was detected in non‐fermenting GNB including Acinetobacter spp. (n = 20), Pseudomonas spp. (n = 11) and Stenotrophomonas maltophila (n = 1). ESBL positive E. coli and carbapenem resistant non‐fermenting GNB are widespread in the backyard poultry flock environment in WA State. These GNB are known to cause opportunistic infections, especially in immunocompromised hosts. Better understanding of the ecology and epidemiology of these GNB in the backyard poultry flock settings is needed to identify potential risks of transmission to people in proximity.     

High level of multidrug-resistant Escherichia coli in young dairy calves in southern Vietnam

This study investigated the occurrence of antimicrobial-resistant Escherichia coli in dairy calves in southern Vietnam. Fecal samples were taken directly from the rectum of 84 calves from 41 smallholder dairy farms, when newborn and at 14 days of age for isolation of E. coli. Escherichia coli strains were isolated from 144 of the 168 fecal samples tested. Of the 144 E. coli isolates, 40% were found to be susceptible to all 12 antimicrobial drugs tested and 53% of the E. coli isolates were resistant to at least three antimicrobials. Calves were colonized with antimicrobial-resistant E. coli already on the day of birth. Resistance to tetracycline was most common, followed by resistance to sulfamethoxazole, ampicillin, trimethoprim, and ciprofloxacin. Four isolates carried a gene encoding for extended-spectrum cephalosporinases (ESC), and these genes belonged to bla_CTX-M group 1 (2 isolates), bla_CTX-M group 9 (1 isolate), and bla_CMY-2 (1 isolate). Thirty-three isolates had a plasmid-mediated quinolone resistance (PMQR) phenotype, and 30 of these carried the qnrS gene. These results are of importance for management routines of dairy cattle to prevent the spread of antimicrobial resistance.

     

A repeated cross‐sectional study of the epidemiology of Campylobacter and antimicrobial resistant Enterobacteriaceae in free‐living Canada geese in Guelph,Ontario, Canada

From May through October 2016, we conducted a repeated cross‐sectional study examining the effects of temporal, spatial, flock and demographic factors (i.e. juvenile vs. adult) on the prevalence of Campylobacter and antimicrobial resistant Enterobacteriaceae among 344 fresh faecal samples collected from Canada geese (Branta canadensis) from four locations where birds nested in Guelph, Ontario, Canada. The overall prevalence of Campylobacter among all fresh faecal samples was 9.3% and was greatest in the fall when these birds became more mobile following the nesting season. Based on 40 gene comparative genomic fingerprinting (CGF40), the increase in prevalence noted in the fall was matched by an increase in the number of unique CGF40 subtypes identified. Resistance to colistin was detected most commonly, in 6% of Escherichia coli isolates, and was highest in the late summer months. All colistin‐resistant isolates were negative for the mcr‐1 to mcr‐5 genes; a chromosomal resistance mechanism (PmrB) was identified in all of these isolates. The prevalence of samples with E. coli exhibiting multi‐class resistance or extended spectrum beta‐lactamase was low (i.e. <2% of samples). The intra‐class correlation coefficients, estimated from the variance components of multilevel logistic regression models, indicated that the shedding of Campylobacter and antimicrobial resistant E. coli among geese within a flock (i.e. birds collected from the same site on the same day) was moderately correlated. Spatial, temporal, and spatiotemporal clusters identified using the spatial scan statistic, largely supported the findings from our multi‐level models. Salmonella was not isolated from any of the fresh faecal samples collected suggesting that its prevalence in this population of birds was very low.     

Genetic Characterization of Antibiotic Resistance in Enteropathogenic Escherichia coli Carrying Extended‐Spectrum β‐Lactamases Recovered from Diarrhoeic Rabbits

A total of 52 Escherichia coli strains isolated from diarrhoeic rabbits were investigated for their enteropathogenic E. coli (EPEC) pathotype by PCR amplification of eae and bfp virulence genes. A total of 22 EPEC isolates were identified, serotyped and studied for antibiotic resistance and screened for the detection of extended‐spectrum β‐lactamases (ESBLs). The EPEC isolates belonged to three serogroups (O26, O92 and O103). The most common serogroup (O103:K‐:H2) was observed among 17 EPEC strains, the O92:K‐serogroup in three isolates (the antibiotic sensitive ones) and the remaining O26:K‐serogroup in two isolates (the ESBLs isolates). Resistances to ampicillin and tetracycline were the most frequent and detected followed by resistance to nalidixic acid, streptomycin, trimethoprim–sulphamethoxazole, cefoxitin, gentamicin and ciprofloxacin. All the isolates were sensitive for amikacin, ceftazidime, aztreonam, imipenem, chloramphenicol, tobramycin and amoxicillin + clavulanic acid. Two isolates recovered from two adult animals showed an intermediate susceptibility to cefotaxime, and a positive screening test for ESBL was demonstrated in both. The bla_TEM gene was demonstrated in the majority of ampicillin‐resistant isolates. The aac(3)‐II or aac(3)‐IV genes were detected in the four gentamicin‐resistant isolates. In addition, the aadA gene was detected in 60% of streptomycin‐resistant isolates. The tet(A) or tet(B) genes were identified in all tetracycline‐resistant isolates. A total of nine EPEC isolates showed the phenotype SXT‐resistant, and the sul1 and/or sul2 and/or sul3 genes were detected in all of them. Our findings showed that the molecular detection by the eae and bfp genes by PCR followed by serotyping is useful for monitoring trends in EPEC infections of rabbits allowing the identification of their possible reservoirs. The detection of genes involved in the resistance to antibiotics of different families in a relatively high proportion of faecal E. coli isolates of rabbits is of great interest and could be considered a serious public health problem.