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This study provides an emended description of Acinetobacter faecalis, a species previously described based on a single isolate (YIM 103518 T ) from elephant feces in China. Our emended description is based on 15 novel isolates conspecific with the A. faecalis type strain, obtained from eight cattle farms in the Czech Republic. The A. faecalis strains have relatively small genomes (≈2.5-2.7 Mbp), with a GC content of 36.3-36.7 mol%. Core genome-based phylogenetic analysis showed that the 15 strains, together with the type strain of A. faecalis, form a distinct and internally coherent phylogroup within the genus. Pairwise genomic ANIb values for the 16 A. faecalis strains were 97.32-99.04 %, while ANIb values between the genomes of the 16 strains and those of the other Acinetobacter spp. were ≤ 86.2 %. Analysis of whole-cell MALDI-TOF mass spectra supported the distinctness and cohesiveness of the taxon. The A. faecalis strains could be differentiated from the other validly named Acinetobacter spp. by the absence of hemolytic activity along with their ability to grow at 37 °C and on L-aspartate, ethanol, and L-glutamate but not at 41 °C or on adipate or 2,3-butanediol. Reduced susceptibility to sulfamethoxazole, trimethoprim and/or streptomycin was shown in eight strains, along with the presence of corresponding antibiotic resistance genes. In conclusion, this study provides a comprehensive description of A. faecalis and demonstrates its occurrence in cattle feces. Though the ecological role of A. faecalis remains unknown, our results show its ability to acquire antibiotic resistance genes, likely as an adaptation to antibiotic selection pressure in livestock farms., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Antimicrobials have been widely used in dairy farms to prevent and control dairy cattle diseases since 1960s. This led to the emergence of antimicrobial resistant bacteria (ARB) that, along with their antimicrobial resistance genes (ARGs), can spread from dairy farms to humans. Therefore, regular antimicrobial resistance (AMR) monitoring is important to implement proper mitigation measures. The objective of this study was to determine the prevalence of AMR and extended-spectrum beta-lactamases (ESBLs)-producing Escherichia coli in dairy cattle. A cross-sectional study was conducted in four dairy cattle farms (A-D) in East Tennessee. A total of 80 samples consisting of 20 samples each of bulk tank milk, feces, dairy cattle manure-amended soil, and prairie soil adjacent to the farms were collected and cultured for the isolation of E. coli . Tetracycline (TET r )-, third-generation cephalosporin (TGC r )- and nalidixic acid (NAL r )-resistant E. coli ( n  = 88) were isolated and identified on agar media supplemented with TET, cefotaxime, and NAL, respectively. TGC r E. coli were tested for ESBLs and other coselected ARGs. TET r (74%, n  = 88) was the most common, followed by TGC r (20%) and NAL r (8%). Farms had significant ( p  < 0.001) differences: the highest prevalence of TGC r (55%) and TET r (100%) were observed in farm D, while all NAL r isolates were from farm C. Over 83% of TGC r isolates ( n  = 18) harbored ESBL gene bla CTX-M . Majority (78%) of the E. coli isolates were multidrug-resistant (MDR), being positive for beta-lactams ( bla CTX-M ), TETs tet ( A ), tet ( B ), tet ( M )), sulfonamides ( sul 2), aminoglycosides ( strA ), and phenicols ( floR ). This study indicated the widespread occurrence of MDR ESBLs- E. coli in dairy cattle farms. AMR surveillance of more dairy farms and identification of farm-level risk factors are important to mitigate the occurrence and spread of ARB of significant public health importance, such as ESBLs- E. coli .

Intestinal carriage of extended spectrum β-lactamase (ESBL)-producing Escherichia coli is a frequent, increasing, and worrying phenomenon, but little is known about the molecular scenario and the evolutionary forces at play. We screened 45 veal calves, known to have high prevalence of carriage, for ESBL-producing E. coli on 514 rectal swabs (one randomly selected colony per sample) collected over 6 months. We characterized the bacterial clones and plasmids carrying bla ESBL genes with a combination of genotyping methods, whole genome sequencing, and conjugation assays. One hundred and seventy-three ESBL-producing E. coli isolates [ bla CTX-M-1 (64.7%), bla CTX-M-14 (33.5%), or bla CTX-M-15 (1.8%)] were detected, belonging to 32 bacterial clones, mostly of phylogroup A. Calves were colonized successively by different clones with a trend in decreasing carriage. The persistence of a clone in a farm was significantly associated with the number of calves colonized. Despite a high diversity of E. coli clones and bla CTX-M -carrying plasmids, few bla CTX-M gene/plasmid/chromosomal background combinations dominated, due to (i) efficient colonization of bacterial clones and/or (ii) successful plasmid spread in various bacterial clones. The scenario "clone versus plasmid spread" depended on the farm. Thus, epistatic interactions between resistance genes, plasmids, and bacterial clones contribute to optimize fitness in specific environments. IMPORTANCE The gut microbiota is the epicenter of the emergence of resistance. Considerable amount of knowledge on the molecular mechanisms of resistance has been accumulated, but the ecological and evolutionary forces at play in nature are less studied. In this context, we performed a field work on temporal intestinal carriage of extended spectrum β-lactamase (ESBL)-producing Escherichia coli in veal farms. Veal calves are animals with one of the highest levels of ESBL producing E. coli fecal carriage, due to early high antibiotic exposure. We were able to show that calves were colonized successively by different ESBL-producing E. coli clones, and that two main scenarios were at play in the spread of bla CTX-M genes among calves: efficient colonization of several calves by a few bacterial clones and successful plasmid spread in various bacterial clones. Such knowledge should help develop new strategies to fight the emergence of antibiotic-resistance.

The number of bacterial genomes deposited each year in public databases is growing exponentially. However, efforts to use these genomes to track trends in antimicrobial resistance (AMR) have been limited thus far. We used 22,102 genomes from public databases to track AMR trends in nontyphoidal Salmonella in food animals in the United States. In 2018, genomes deposited in public databases carried genes conferring resistance, on average, to 2.08 antimicrobial classes in poultry, 1.74 in bovines, and 1.28 in swine. This represents a decline in AMR of over 70% compared to the levels in 2000 in bovines and swine, and an increase of 13% for poultry. Trends in resistance inferred from genomic data showed good agreement with U.S. phenotypic surveillance data (weighted mean absolute difference ± standard deviation, 5.86% ± 8.11%). In 2018, resistance to 3rd-generation cephalosporins in bovines, swine, and poultry decreased to 9.97% on average, whereas in quinolones and 4th-generation cephalosporins, resistance increased to 12.53% and 3.87%, respectively. This was concomitant with a decrease of bla CMY-2 but an increase in bla CTX-M-65 and gyrA D87Y (encoding a change of D to Y at position 87). Core genome single-nucleotide polymorphism (SNP) phylogenies show that resistance to these antimicrobial classes was predominantly associated with Salmonella enterica serovar Infantis and, to a lesser extent, S. enterica serovar Typhimurium and its monophasic variant I 4,[5],12:i:-, whereas quinolone resistance was also associated with S. enterica serovar Dublin. Between 2000 and 2018, trends in serovar prevalence showed a composition shift where S. Typhimurium decreased while S. Infantis increased. Our findings illustrate the growing potential of using genomes in public databases to track AMR in regions where sequencing capacities are currently expanding. IMPORTANCE Next-generation sequencing has led to an exponential increase in the number of genomes deposited in public repositories. This growing volume of information presents opportunities to track the prevalence of genes conferring antimicrobial resistance (AMR), a growing threat to the health of humans and animals. Using 22,102 public genomes, we estimated that the prevalence of multidrug resistance (MDR) in the United States decreased in nontyphoidal Salmonella isolates recovered from bovines and swine between 2000 and 2018, whereas it increased in poultry. These trends are consistent with those detected by national surveillance systems that monitor resistance using phenotypic testing. However, using genomes, we identified that genes conferring resistance to critically important antimicrobials were associated with specific MDR serovars that could be the focus for future interventions. Our analysis illustrates the growing potential of public repositories to monitor AMR trends and shows that similar efforts could soon be carried out in other regions where genomic surveillance is increasing.

Campylobacter is one of the most common causes of foodborne bacterial infections worldwide. Why poultry has been shown to be one of the most significant sources of these bacteria, ruminants, especially cattle, are also responsible for a high number of human Campylobacter jejuni , and to a lesser extent Campylobacter coli , infections. In this study, bovine and pig carcasses in Poland were investigated for the presence of Campylobacter and for their antimicrobial resistance. A total of 204 swabs from bovine carcasses and 355 swab samples from pig carcasses were tested during 2014-2018. Campylobacter was identified in 129 (36.3%) of the pig and in 11 (5.4%) of the bovine carcasses, respectively. The pig isolates were classified as C. coli (121; 34.1%) or C. jejuni (8; 2.3%), whereas the bovine Campylobacter were identified either as C. jejuni (8; 3.9% isolates) or C. coli (3; 1.5% strains). Resistance of the isolates ( n  = 140) to erythromycin, ciprofloxacin, nalidixic acid, streptomycin, and tetracycline revealed that the vast majority of C. coli was resistant to streptomycin (106 isolates; 85.5%), tetracycline (97; 78.2%), nalidixic acid (90; 72.6%), and ciprofloxacin (88; 71.0%). Among C. jejuni isolates ( n  = 16) the resistance rates to all antibiotics were lower than in C. coli , irrespective of the origin. A total of 74 of 121 (61.2%) C. coli isolates from the pig carcasses and one of three such isolates from the bovine samples were multiresistant. Most of the C. coli (64 isolates; 85.3%) had the ciprofloxacin+nalidixic acid+streptomycin+tetracycline resistance profile. The results suggest that pig and bovine carcasses may be an underestimated reservoir of Campylobacter , especially for C. coli in pigs. The high antimicrobial resistance rates of such strains to streptomycin, quinolones, and tetracyclines highlight the need for monitoring of these bacteria in such food and food products.

Colistin is a last-resort antimicrobial used to treat infections caused by multidrug-resistant Gram-negative bacilli (MDR-GNB). The emergence of colistin resistance, particularly linked to mobile genetic elements including the mcr genes, is a major threat to the management of MDR-GNB infections. The aim of this study was to assess the presence of mcr genes in a collection of 40 colistin-resistant commensal Escherichia coli isolated from healthy pigs, cattle and poultry in Belgium between 2012 and 2016. All isolates carried at least one mcr gene. The genes mcr-1 to -5 were observed in this collection. Different replicons associated with mcr genes were identified, including IncHI2/IncHI2A associated with mcr-1, IncX4 associated with mcr-1 and mcr-2, and ColE10 associated with mcr-4. While the occurrence of multiple mcr genes in a single isolate has rarely been reported elsewhere, a triple occurrence (mcr-1, -3 and -5) was found in this study. All isolates were MDR and carried between one and nine different replicons. Seventeen different sequence types were observed among the 40 E. coli isolates. In conclusion, this study revealed the presence of a reservoir of mobile colistin resistance genes (mcr-1 to -5) observed during at least 5 years (2012-2016) in the commensal gut flora of pigs, cattle and poultry in Belgium., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Selection pressure exerted by use of antibiotics in both human and veterinary medicine is responsible for increasing antimicrobial resistance (AMR). The objectives of this study were to better understand antimicrobial use in pigs, beef cattle, and poultry on farms on Guadeloupe, French West Indies, and to acquire data on AMR in Escherichia coli in these food-producing animals. A cross-sectional survey was conducted at 45 farms on Guadeloupe, and practical use of antimicrobials was documented in declarative interviews between March and July 2018. A total of 216 fecal samples were collected between January 2018 and May 2019, comprising 124 from pigs, 75 from beef cattle, and 17 from poultry litter. E. coli isolates were obtained for further testing by isolation and identification from field samples. Antimicrobial susceptibility testing and screening for bla CTX-M , bla TEM , tetA, and tetB resistance genes by polymerase chain reaction on extracted genomic DNA were performed., Results: The study showed rational use of antimicrobials, consisting of occasional use for curative treatment by veterinary prescription. Tetracycline was the most commonly used antimicrobial, but its use was not correlated to E. coli resistance. Extended-spectrum β-lactamase (ESBL) E. coli isolates were detected in 7.3% of pigs, 14.7% of beef cattle, and 35.3% of poultry. bla CTX-M-1 was the predominant gene found in ESBL-E. coli isolates (68.8%), followed by bla CTX-M-15 (31.3%)., Conclusion: Despite rational use of antimicrobials, the rate of ESBL-E. coli in food-producing animals in Guadeloupe, although moderate, is a concern. Further studies are in progress to better define the genetic background of the ESBL-E. coli isolates.

Antimicrobial resistance has become a global threat to public health since multidrug-resistant (MDR) bacteria have been reported worldwide carrying different antimicrobial resistance genes (ARGs), and animals have been described as a reservoir of ARGs. The presence of antimicrobial-resistant bacteria and ARGs in the food matrix is a risk to public health. This study aimed to research the presence of clinically relevant ARGs for important antimicrobials and genetic elements in fecal samples from dairy cows and calves on a Brazilian farm. In this study, a total of 21 fecal samples were collected, and then, the DNA of cultivable aerobic bacteria was extracted. Fifty-seven ARGs and twenty-three genetic elements were researched by PCR and confirmed by sequencing. Several ARGs that confer resistance to β-lactams, tetracyclines, fluoroquinolones, sulphonamides, phenicols, aminoglycoside, glycopeptides, and macrolides were detected. A total of 200 amplicons from 23 ARGs (bla CTX-M-Gp2 , bla CMY , bla SHV , tetA, tetB, tetC, qepA, qnrB, qnrS, oqxA, oqxB, vanC1, vanC2/3, aadA, sul1, sul2, sul3, ermB, mefAE, floR, cmlA, aadA, aph(3')-Ia, aac(3')-Ia), and 145 amplicons from 12 genetic elements (IncF, IncFIA, IncFIB, IncI1, IncY, IncU, IncK, IncP, IncR, IncHI1, ColE-like, intI1) were detected. The results presented in this study call attention to the monitoring of antimicrobial resistance in dairy farms worldwide. MDR bacteria and ARGs can spread to different sources, including milk products, which are one of the most consumed products worldwide, representing a potential risk to human health.

This study characterized Listeria monocytogenes, Salmonella enterica, and E. coli O157:H7 by collecting rectoanal mucosal swabs and fecal samples from 518 imported beef cattle at Jordan's major abattoir. A unique 53 L. monocytogenes, 287 S. enterica, and 17 E. coli O157:H7 were isolated from 37, 120 and 9 different animals; respectively. The prevalence of S. enterica, L. monocytogenes and E. coli O157:H7 were 23.2 % (95 % CI, 19.7-27.0 %), 7.1 % (95 % CI, 5.2-9.7 %) and 1.7 % (95 % CI, 0.9-3.3 %); respectively. All L. monocytogenes, all E. coli O157:H7 and 93.0 % of S. enterica isolates resisted at least one antimicrobial class. All L. monocytogenes, 94.1 % of E. coli O157:H7 and 69.7 % of S. enterica isolates exhibited multidrug resistance (resistant to ≥3 antimicrobials classes). Moreover, high percentages of L. monocytogenes (98.1 %), E. coli O157:H7 (64.7 %) and S. enterica (45.3 %) isolates resisted ≥5 antimicrobial classes. More than 90 % of the L. monocytogenes isolates resisted ampicillin, penicillin and erythromycin and more than 75 % resisted vancomycin. S. enterica isolates resisted several treatment-of-choice antimicrobials such as nalidixic acid (85.4 %), ciprofloxacin (26.8 %) and ceftriaxone (19.5 %). Furthermore, greater than 50 % of the E. coli O157:H7 isolates resisted streptomycin, nalidixic acid, tetracycline, ampicillin, sulfamethoxazole-trimethoprim, kanamycin, chloramphenicol and ciprofloxacin. The high prevalence and the high resistance percentages of the studied pathogens toward clinically important antimicrobials is alarming. Thus, applying strict sanitation procedures at the abattoirs in Jordan is crucial to lower the risk of carcasses contamination., Competing Interests: Declaration of Competing Interest The author declares no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The aim of this article is to report on antimicrobial resistance (AMR) in commensal Escherichia coli from livestock from several European countries. The relationships with antimicrobial usage (AMU) at country level and harmonized indicators to cover the most relevant AMR aspects for human health in animal production were also investigated. E. coli were isolated in faeces from broilers and fattening pigs (from nine countries), and fattening turkeys and veal calves (from three countries) and screened against a fixed antimicrobial panel. AMU data were collected at farm and average treatment incidences stratified by antimicrobial class, country and livestock species were calculated. Associations between AMR and AMU at country level were analysed. Independent of animal species, the highest resistance was observed for ampicillin, sulphamethoxazole, tetracycline and trimethoprim. E. coli from broilers showed the highest resistance level for (fluoro)quinolones, and multidrug resistance peaked in broilers and fattening turkeys. Colistin resistance was observed at very low levels with the exception of fattening turkeys. High resistance to third- and fourth-generation cephalosporins was detected in broilers and fattening turkeys. The lowest levels of resistance were for meropenem, azithromycin and tigecycline (<1 %). Significant correlations between resistance and usage at country level were detected in broilers for polymyxins and aminoglycosides, and in fattening pigs for cephalosporins, amphenicols, fluoroquinolones and polymyxins. None of the correlations observed between AMR and AMU were statistically significant for fattening turkey and veal calves. The strength of the analysis performed here is the correlation of aggregated data from the same farms at country level for both AMU and AMR within antimicrobial classes.

The purpose of this study was to determine the effect of intramammary pirlimycin on the fecal microbiome of dairy cattle. Primiparous heifers were enrolled and assigned to a treatment or control group at a ratio of 2:1. In part 1 of the study, treated heifers (T1) were given intramammary pirlimycin into one infected quarter once daily for 2 d at 24-h intervals, according to the label instructions. Control heifers received no treatment. In part 2 of the study, treated heifers (T2) were given intramammary pirlimycin into one infected quarter once daily for 8 d at 24-h intervals, according to the label instructions. All enrolled heifers (T1, T2, and control) had quarter-level milk samples aseptically collected for bacterial culture and fecal samples collected for 16S rRNA gene sequencing on d 0, 2, 7, 14, 21, and 28. Milk samples were plated on Columbia blood agar and incubated at 37°C for 24 h. Bacteria were identified using MALDI-TOF mass spectrometry. The DNA was extracted from feces using PowerFecal kits (Qiagen, Venlo, the Netherlands). The 16S rRNA gene amplicon library construction and sequencing was performed at the University of Missouri DNA Core facility. Testing for differences in fecal community composition was performed via one-way permutational multivariate ANOVA of Bray-Curtis and Jaccard similarities using Past 3.13 (https://folk.uio.no/ohammer/past/). Mean total count of operational taxonomic units and Chao1, Shannon, and Simpson α-diversity indices were determined and compared via t-test or Wilcoxon rank sum test. A treatment-dependent effect was present in the observed and predicted richness of feces from cows in the T1 group at d 2 posttreatment. Additionally, intramammary pirlimycin induced a significant change in the composition of the fecal microbiota by d 2 in the treated groups. Based on calculated intra-subject similarities, intramammary pirlimycin was associated with a significant acute change in the fecal microbiota of dairy heifers and that chance reversed when the antimicrobial exposure was brief, but sustained following longer exposure. Overall, intramammary pirlimycin administration affected the fecal microbiome of lactating dairy heifers. Further work is necessary to determine the effect of these changes on the heifer and the dairy environment as well as if treatment is influencing antimicrobial resistance among enteric and environmental bacteria., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Antimicrobial resistance reported in bacteria of animal origin is considered a major challenge to veterinary public health. In this study, the genotypic and phenotypic characterisation of twelve Escherichia coli isolates of bovine origin is reported. Twelve bacterial isolates of animal origin were selected from a previous study based on their multidrug resistant (MDR) profile. Efflux pump activity was measured using ethidium bromide (EtBr) and the biofilm forming ability of the individual strains was assessed using a number of phenotypic assays. All isolates were resistant to tetracyclines and a number of isolates expressed resistance to fluoroquinolones which was also confirmed in silico by the presence of these resistance markers. Amino acid substitutions in the quinolone resistance-determining regions were identified in all isolates and the presence of several siderophores were also noted. Whole genomesequence (WGS) data showed different STs that were not associated with epidemic STs or virulent clonal complexes. Seven isolates formed biofilms in minimal media with some isolates showing better adaptation at 25 °C while others at 37 °C. The capacity to efflux EtBr was found to be high in 4 isolates and impaired in 4 others. The pathogenicity of three selected isolates was assessed in zebrafish embryo infection models, revealing isolates CFS0355 and CFS0356 as highly pathogenic. These results highlight the application of NGS technologies combined with phenotypic assays in providing a better understanding of E. coli of bovine origin and their adaptation to this niche environment., Competing Interests: Declaration of Competing Interest The authors have nothing to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Antimicrobial resistance is a threat to agricultural production and public health. In this proof-of-concept study, we investigated predicting antimicrobial sensitive/resistant (S/R) phenotypes and host sources of Escherichia coli (n = 128) based on differential fatty acid abundance. Myristic (14:0), pentadecanoic acid (15:0), palmitic (16:0), elaidic (18:1 9 ) and steric acid (18:0) were significantly different (α = 0.05) using a two-way ANOVA for predicting nalidixic acid, ciprofloxacin, aztreonam, cefatoxime, and ceftazidime S/R phenotypes. Additionally, analyses of palmitoleic (16:1), palmitic acid (16:0), methyl palmitate (i-17:0), and cis-9,10-methyleneoctadecanoic acid (19:0 Δ ) showed these markers were significantly different (α = 0.05) between isolates obtained from cattle and raccoons. S/R phenotype prediction for the above antibiotics or host source, based on linear regression models of fatty acid abundance, were made using a replicated-randomized subsampling and modeling approach. This model predicted S/R phenotype with 79% and 81% accuracy for nalidixic acid and ciprofloxacin, respectively. The isolate host source was predicted with 63% accuracy., (Published by Elsevier Inc.)

A total of 96 Trueperella pyogenes isolates, an opportunistic pathogen of food-producing ruminants, obtained from cattle (n = 34), sheep (n = 35) and goats (n = 27), and identified by Real Time PCR (qPCR), were analysed to determine the susceptibility to 12 antimicrobials commonly used in livestock, using a broth microdilution. The Minimal Inhibitory Concentration (MIC) distribution was unimodal for half of the antimicrobials tested with the exception of apramycin, gentamicin, streptomycin, oxytetracycline, tylosin, and erythromycin all of which showed bimodal MIC distributions. Low MIC 90 values for penicillin, amoxicillin, ceftiofur, enrofloxacin, and gentamicin (<1 μg/ml) were obtained, suggesting that these antimicrobials would be the most effective first line empiric treatment for T. pyogenes infections in livestock. Furthermore, according to the specific T. pyogenes breakpoints for penicillin, sulfamethoxazole/trimethoprim and erythromycin, 93.7 % of isolates were susceptible to penicillin and 77.2 % to erythromycin, whereas 92.7 % were non-susceptible to sulfamethoxazole/trimethoprim. Significant differences were observed in the MIC distribution of almost all antimicrobials, except enrofloxacin, tylosin and erythromycin against cattle, sheep or goat isolates, although all antimicrobials showed similar MIC 90 values, except apramycin and oxytetracycline that showed higher values when tested against cattle isolates. These data provide interesting information on the antimicrobials of choice for the treatment of infections caused by T. pyogenes in ruminants., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Methicillin resistant S. aureus from cows with mastitis has received a growing interest worldwide. The present study aimed to provide a detailed description of the resistance and virulence traits of isolates from bovine mastitis samples. A total of 550 quarter milk samples were collected from 140 mastitic household dairy cows and buffalo from five herds at Dakahlia Governorate, Egypt, during 2017 and 2018. Staphylococcus spp. were isolated and differentiated using MALDI-TOF MS. A genotypic characterization was performed for S. aureus isolates using DNA-microarray and staphylococcal protein A (spa) typing. Furthermore, antibiotic resistances were phenotypically confirmed using broth microdilution. Six different clonal lineages (CC1-MRSA, CC5-MRSA, CC45-MRSA, CC97-MSSA, CC50-MSSA and CC1153-MSSA), including seven spa types (t127, t688, t132, t267, t521, t224 and t903) were identified. Spa type t267 was the most dominant among the investigated herds. This is the first report of the occurrence of clonal lineages CC97, CC1, CC45, CC50 and CC1153 from bovine mastitis in Egypt. All MRSA isolates and 33.3 % of MSSA were multi-resistant (i.e. resistant to more than three classes of compounds). Various virulence determinants were also observed including leukocidins, hemolysins, and enterotoxins. The study demonstrates a low diversity of S. aureus isolates recovered from several dairy herds. The findings of the observed virulotypes can be useful for future studies on anti-virulence therapies, immunogenicity and vaccine development., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Antimicrobial resistance is a relevant "One Health" issue that shows the need of comparison of isolates of different origins. In this way, guidelines for antimicrobial-resistance evaluation in animals are relevant in relation to human sources. This work aims to compare antimicrobial-resistance results of animal isolates considering CLSI and EUCAST guidelines. The comparison shows considerable differences in the results, which include antibiotics used as primary options in hospital infections. EUCAST showed the higher number of samples with resistance profiles than CLSI that indicates a more efficient scenario to the EUCAST to screen antibiotic-resistant bacteria. EUCAST was more consonant to the expected phenotype for ESBL producers, with higher index of resistance to oxyimino-beta-lactam antibiotics. The study shows that there are differences in the interpretative results using different guidelines, where the susceptibility test results concerning Enterobacteriaceae of animal origin are not always coincident in CLSI and EUCAST. EUCAST has proved to be the most reliable alternative for profile screening of antibiotic resistance, when compared to CLSI. We might say the same with respect to the ESBL-producing Enterobacteriaceae , in which EUCAST has proved to be more efficient about the demonstration of expected resistance profiles for the ESBL producers. These differences show that guideline selection might influence the therapeutic option.

The aims of this study were to investigate the plasmid-mediated colistin resistance genes ( mcr-1 , mcr-2 , mcr-3 , mcr-4 , and mcr-5 ), phenotypic colistin resistance in Escherichia coli O157:H7 + /H7 - strains isolated from cattle and sheep, and whole-genome sequence (WGS) analysis of colistin-resistant sorbitol fermentative E. coli O157:H7. According to the results, 5 of the 49 isolates were found to harbor mcr-2 and/or mcr-3 genes. Three isolates, including a sorbitol fermentative E. coli O157:H7, were found phenotypically resistant to colistin with a minimum inhibitory concentration value of 128 μg/mL. The genome of sorbitol fermentative E. coli O157:H7 did not show 100% similarity to any of the other genome sequences found in the universal genome database. It has also been determined that this isolate carried 62 different antimicrobial resistance genes. This is the first report of plasmid-mediated mcr-2 and mcr-3 genes carrying E. coli O157:H7 from cattle and sheep isolates and WGS of a colistin-resistant sorbitol fermentative E. coli O157:H7. Findings of this study indicate that cattle and sheep can be an important source of colistin resistance in E. coli O157:H7, and slaughterhouse wastewater might be a significant route for dissemination of the plasmid-mediated colistin genes. Therefore, the use of colistin in veterinary medicine should be restricted to reduce the development of resistance. Also it may be necessary to review the non-sorbitol fermentation-based isolation protocol for not missing the sorbitol fermentative E. coli O157:H7 in epidemiological studies.

Outbreaks of Salmonellosis remain a major public health problem globally. This study determined the diversity and antibiotic resistance gene profile of Salmonella enterica serovars isolated from humans and food animals. Using standard methods, Salmonella spp. were isolated from fecal samples, profiled for antimicrobial susceptibility and resistance genes. Seventy-one Salmonella isolates were recovered from both humans and food animals comprising cattle, sheep, and chicken. Forty-four serovars were identified, with dominant Salmonella Budapest (31.8%). Rare serovars were present in chicken ( S . Alfort, S . Wichita, S . Linton, S . Ealing, and S . Ebrie) and humans ( S . Mowanjum, S . Huettwillen, S . Limete, and S . Chagoua). Sixty-eight percent of isolates were sensitive to all test antibiotics, while the highest rate of resistance was to nalidixic acid (16.9%; n  = 12), followed by ciprofloxacin (11.3%; n  = 8) and tetracycline (9.9%; n  = 8). Five isolates (7%) were multidrug-resistant and antimicrobial resistance genes coding resistance to tetracycline ( tetA ), beta-lactam ( bla TEM ), and quinolone/fluoroquinolone ( qnrB and qnrS ) were detected. Evolutionary analysis of gyrA gene sequences of human and food animal Salmonella isolates revealed variations but are evolutionarily interconnected. Isolates were grouped into four clades with S . Budapest isolate from cattle clustering with S . Budapest isolated from chicken, whereas S . Essen isolated from sheep and chicken was grouped into a clade. Diverse S. enterica serovars with high antibiotic resistance profile are ubiquitous in food animals; hence, there is a need for surveillance and prudent use of antibiotics in human and veterinary medicine.

Objective: To determine the prevalence and genetic relatedness of bla CTX-M -type extended spectrum-β-lactamase (ESBL)-producing Escherichia coli at the human-animal interface in Pakistan. Materials and Methods: A total of 150 human, cattle, and poultry fecal samples (50 each) were screened for ESBL-producing E. coli using ESBL CHROMagar ® . Bacterial species confirmation as well as determination of minimum inhibitory concentrations (μg/mL) to different antibiotics was performed using the automated VITEK ® -2 compact system. Phenotypic confirmation of ESBL production was performed according to the Clinical Laboratory Standards Institute (CLSI) guidelines. Genetic analysis of bla CTX-M was carried out by PCR and DNA sequencing. Plasmids and clonal similarity of the E. coli strains were determined by PCR-based replicon typing and pulsed-field gel electrophoresis (PFGE), respectively. Results: Of 150 samples, 29 (19.3%) ESBL-producing E. coli were recovered, and majority of them originated from human ( n  = 16; 55%), followed by cattle ( n  = 9; 31%) and poultry ( n  = 4; 13.7%). bla CTX-M-15 was predominant ESBL genotype ( n  = 25; 86.2%), mainly identified from human ( n  = 15) and cattle ( n  = 9). This is also the first report of the occurrence of CTX-M-15 and CTX-M-55 in cattle and poultry E. coli isolates of Pakistan, respectively. The majority of the ESBL-producing E. coli (96.5%) showed a multidrug resistance phenotype. All isolates carried IncFII or IncFIA plasmids, and the phylogroup B1 was dominant (44.8%) followed by phylogroups A (31%), D (17.2%), and B2 (6.8%). PFGE revealed that isolates from different hosts were genetically unrelated. Conclusion: Presence of CTX-M-15-type ESBL-producing E. coli in different reservoirs is alarming and has the potential to impact both veterinary and human therapeutic treatment options.

This study determined the prevalence, pulsed-field gel electrophoresis profiles, and antimicrobial resistance profile of Listeria monocytogenes, Salmonella enterica, and Escherichia coli O157:H7 isolates from dairy cattle farms in Jordan. Samples from bulk tank milk (n = 305), cattle feces (n = 610), and rectoanal mucosal swabs (n = 610) were collected from 61 dairy cattle farms. We confirmed 32 L. monocytogenes, 28 S. enterica, and 24 E. coli O157:H7 isolates from the samples. The farm-level prevalence (at least 1 positive sample per farm) of L. monocytogenes, S. enterica, and E. coli O157:H7 was 27.9, 19.7, and 23.0%, respectively. The prevalence of L. monocytogenes, S. enterica, and E. coli O157:H7 in bulk tank milk was 7.5, 1.6, and 3.3%, respectively. The prevalence of L. monocytogenes and S. enterica in fecal samples was 1.5 and 3.8%, respectively, and the prevalence of E. coli O157:H7 in rectoanal mucosal swabs was 2.3%. Based on disk diffusion testing, all L. monocytogenes, S. enterica, and E. coli O157:H7 isolates exhibited resistance to at least 1 antimicrobial class. Multidrug resistance (resistance to 3 or more classes of antimicrobials) was exhibited by 96.9% of L. monocytogenes, 91.7% of E. coli O157:H7, and 82.1% of S. enterica isolates. Moreover, 93.8, 79.2, and 57.1% of the L. monocytogenes, E. coli O157:H7, and S. enterica isolates, respectively, were resistant to 5 or more antimicrobial classes. More than 50% of L. monocytogenes isolates were resistant to ampicillin, clindamycin, penicillin, erythromycin, quinupristin-dalfopristin, streptomycin, teicoplanin, linezolid, vancomycin, kanamycin, and tetracycline. More than 50% of S. enterica and E. coli O157:H7 isolates were resistant to ampicillin, cephalothin, nalidixic acid, kanamycin, streptomycin, amoxicillin-clavulanic acid, and tetracycline. The prevalence of the studied pathogens this study was comparable to reports from other countries. The isolated pathogens exhibited a high degree of antimicrobial resistance, suggesting that the bacterial flora of dairy cattle in Jordan are under intense antimicrobial selection pressure. Additional research is required to determine the causes and drivers of resistance, and to develop approaches to mitigating antimicrobial resistance., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Salmonellais a foodborne zoonotic enteric bacterium able to infect both humans and animals. This study aimed to identify the antimicrobial resistance of Salmonella serovars isolated from human, cattle, and poultry. Moreover, we investigated the probable transmission trends of antibiotic-resistant Salmonella isolates from food animals to human. A total of 242 Salmonella isolates collected from various human and animal sources were serotyped. The polymerase chain reaction was performed to detect the invA virulence gene. The isolates were subsequently tested against 14 antimicrobials and the resistance rates among the isolates from three sample sources were statistically analyzed by the Chi-Square test. Serotyping revealed the isolates belonged to various serovars with the dominance of Enteritidis (37%), Typhimurium (35.3%), and Infantis (21.1%). A high frequency of resistance to streptomycin was observed followed by tetracycline, trimethoprim, sulfonamides, spectinomycin, chloramphenicol, florfenicol, ampicillin, kanamycin, ceftazidime, and cefepime. In addition, multidrug resistance was observed in more than 40% of the isolates. The results of the statistical analysis showed a significant relationship (P ˂ 0.001) between the rate of antibiotic resistance among the three sources of Salmonellaisolates. Furthermore, the antibiotic resistance had a statistical relationship between the different serotypes isolated from different sources. These findings demonstrate the possible transmission of resistance to human from animal sources. The prevalence of the Typhimurium, Enteritidis, and Infantis serovars in both human and animals suggested that Salmonella contamination in chicken and cattle may be the major source of salmonellosis in human. The high incidence of antibiotic resistance in Salmonellaisolates along with the close relationship between the antimicrobial resistance of animal and human isolates indicate the role of food animal products as an important source of resistance., (Copyright © 2019, Archives of Razi Institute. Published by Kowsar.)

In this study, 140 cattle STEC isolates belonging to serogroups O157, O26, O145, O121, O103 and O45 were characterized for 38 virulence-associated genes, antimicrobial resistance profiles and genotyped by PFGE. The majority of isolates carried both stx1 and stx2 concurrently, stx2c, and stx2d; plasmid-encoded genes ehxA, espP, subA and saa but lacked katP and etpD and eaeA. Possession of eaeA was significantly associated with the presence of nle genes, katP, etpD, ureC and terC. However, saa and subA, stx1c and stx1d were only detected in eaeA negative isolates. A complete OI-122 and most non-LEE effector genes were detected in only two eaeA positive serotypes, including STEC O157:H7 and O103:H2. The eaeA gene was detected in STEC serotypes that are commonly implicated in severe humans disease and outbreaks including STEC O157:H7, STEC O145:H28 and O103:H2. PFGE revealed that the isolates were highly diverse with very low rates of antimicrobial resistance. In conclusion, only a small number of cattle STEC serotypes that possessed eaeA, had the highest number of virulence-associated genes, indicative of their high virulence. Further characterization of STEC O157:H7, STEC O145:H28 and O103:H2 using whole genome sequencing will be needed to fully understand their virulence potential for humans.

Bovine Respiratory Disease (BRD) is a major threat to animal health and welfare in the cattle industry. Strains of Mannheimia haemolytica (Mh) that are resistant to multiple classes of antimicrobials are becoming a major concern in the beef industry, as the frequency of isolation of these strains has been increasing. Mobile genetic elements, such as integrative conjugative elements (ICE), are frequently implicated in this rapid increase in multi-drug resistance. The objectives of the current study were to determine the genetic relationship between the isolates collected at arrival before metaphylaxis and at revaccination after metaphylaxis, to identify which resistance genes might be present in these isolates, and to determine if they were carried on an ICE. Twenty calves culture positive for Mh at arrival and revaccination were identified, and a total of 48 isolates with unique susceptibility profiles (26 from arrival, and 22 from revaccination) were submitted for whole-genome sequencing (WGS). A phylogenetic tree was constructed, showing the arrival isolates falling into four clades, and all revaccination isolates within one clade. All revaccination isolates, and one arrival isolate, were positive for the presence of an ICE. Three different ICEs with resistance gene modules were identified. The resistance genes aphA1, strA, strB, sul2, floR, erm42, tetH/R, aadB, aadA25, blaOXA-2, msrE, mphE were all located within an ICE. The gene bla-ROB1 was also present in the isolates, but was not located within an ICE., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Surveillance of Pasteurella multocida resistance in food-producing animals is essential to guide the first-line treatment of respiratory diseases and to limit economic losses. Since Pasteurella are the most common bacteria isolated from dog and cat bites, this surveillance is also needed to guide treatment in humans in case of bites. The aim of this study was to characterize the phenotypic resistance of P. multocida strains isolated from respiratory infections in animals, including both food-producing animals and pets. Data collected between 2012 and 2017 by the French national surveillance network for antimicrobial resistance referred to as RESAPATH were analyzed. The proportions of resistance to antimicrobials of relevance in veterinary and human medicines were estimated for each animal species. For cattle, resistance trends over the period were investigated using non-linear analysis applied to time-series. In total, 5356 P. multocida isolates were analyzed. Proportions of resistance of P. multocida were almost all below 20% over the period, and, more precisely, all resistance proportions were below 10% for rabbits, sheep and dogs. The highest resistance proportions to enrofloxacin were identified for cattle (4.5%) and dogs (5.2%). Despite its frequent use in livestock, resistance to florfenicol was less than 1% in P. multocida strains, regardless of the animal species considered. Time series analyses revealed continuous increases in resistance to tetracycline, tilmicosin, flumequine and fluoroquinolones in P. multocida strains isolated from cattle. These trends contrast with the decrease in use of antibiotics in cattle in France and with the decrease in resistance observed in E. coli isolated from diseased cattle., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Genomic analyses were performed on florfenicol-resistant (FFN r ) Campylobacter coli isolates recovered from cattle, and the cfr (C) gene-associated multidrug resistance (MDR) plasmid was characterized. Sixteen FFN r C. coli isolates recovered between 2013 and 2018 from beef cattle were sequenced using MiSeq. Genomes and plasmids were found to be closed for three of the isolates using the PacBio system. Single nucleotide polymorphisms (SNPs) across the genome and the structures of MDR plasmids were investigated. Conjugation experiments were performed to determine the transferability of cfr (C)-associated MDR plasmids. The spectrum of resistance encoded by the cfr (C) gene was further investigated by agar dilution antimicrobial susceptibility testing. All 16 FFN r isolates were MDR and exhibited coresistance to ciprofloxacin, nalidixic acid, clindamycin, and tetracycline. All isolates shared the same resistance genotype, carrying aph (3')-III , hph , ΔaadE (truncated), bla OXA-61 , cfr (C), and tet (O) genes plus a mutation of GyrA (T86I). The cfr (C), aph (3')-III , hph , ΔaadE , and tet (O) genes were colocated on transferable MDR plasmids ranging in size from 48 to 50 kb. These plasmids showed high sequence homology with the pTet plasmid and carried several Campylobacter virulence genes, including virB2 , virB4 , virB5 , VirB6 , virB7 , virB8 , virb9 , virB10 , virB11 , and virD4 The cfr (C) gene conferred resistance to florfenicol (8 to 32 μg/ml), clindamycin (512 to 1,024 μg/ml), linezolid (128 to 512 μg/ml), and tiamulin (1,024 μg/ml). Phylogenetic analysis showed SNP differences ranging from 11 to 2,248 SNPs among the 16 isolates. The results showed that the cfr (C) gene located in the conjugative pTet MDR/virulence plasmid is present in diverse strains, where it confers high levels of resistance to several antimicrobials, including linezolid, a critical drug for treating infections by Gram-positive bacteria in humans. This report highlights the power of genomic antimicrobial resistance surveillance to uncover the intricacies of transmissible coresistance and provides information that is needed for accurate risk assessment and mitigation strategies. IMPORTANCE Campylobacter is a leading cause of foodborne diarrheal illness worldwide, with more than one million cases each year in the United States alone. The global emergence of antimicrobial resistance in this pathogen has become a growing public health concern. Florfenicol-resistant (FFN r ) Campylobacter has been very rare in the United States. In this study, we employed whole-genome sequencing to characterize 16 multidrug-resistant Campylobacter coli isolates recovered from cattle in the United States. A gene [ cfr (C)] was found to be responsible for resistance not only to florfenicol but also to several other antimicrobials, including linezolid, a critical drug for treating infections by Gram-positive bacteria in humans. The results showed that cfr (C) is located in a conjugative pTet MDR/virulence plasmid. This report highlights the power of antimicrobial resistance surveillance to uncover the intricacies of transmissible coresistance and provides information that is needed for accurate risk assessment and mitigation strategies.

Background: Beef cattle in North America frequently receive an antibiotic injection after feedlot placement to control and manage bovine respiratory disease. The potential collateral effect of these antibiotics on the bovine microbiome is largely unknown. Therefore, we determined the longitudinal impact of two commonly administered veterinary antibiotics, oxytetracycline and tulathromycin, on the fecal and nasopharyngeal (NP) microbiota of beef cattle that were transported to a feedlot. We also report the effect these antibiotics have on several antibiotic resistance determinants in both the fecal and NP microbiome., Results: Oxytetracycline and tulathromycin perturbation of the bovine fecal and NP microbiota was greatest at days 2 and 5. Although the NP microbiota of the tulathromycin-treated cattle had recovered by day 12, the NP microbiota of the oxytetracycline-treated group remained altered through day 34. Overall, the NP microbiota appeared to be more sensitive to antibiotic treatment than the fecal microbiota. Members of the bacterial Microbacteriaceae family were most notably affected by antibiotic administration in the NP microbiota. Both antibiotics protected against Pasteurella spp. in the nasopharynx at days 2 and 5. Despite very similar diets at both locations, the largest shift in the fecal and NP microbiota occurred after transport to the feedlot (P < 0.05). Antibiotic resistance determinants in the NP microbiome were also affected more strongly by antibiotic treatment than those in the fecal microbiome. Oxytetracycline increased the proportion of erm(X), sul2, tet(H), tet(M), and tet(W) in NP samples and tet(M) and tet(W) in fecal samples, at day 12 (P < 0.05). The effect of tulathromycin on the relative abundance of resistance genes in the NP microbiome was greatest at day 34 as erm(X), sul2, and tet(M) were enriched (P < 0.05)., Conclusions: Administration of a single injection of oxytetracycline and tulathromycin resulted in significant changes in the NP and fecal microbiota during the first 5 days after treatment. Antibiotic treatment also increased the relative abundance of several antibiotic resistance determinants in the fecal and NP microbiome at either day 12 or 34.

Antibiotics can truly be considered the symbol of the 20th century's miracle drugs. However, excessive and inappropriate use of antibiotics in human and veterinary medicine has led to repercussions in terms of mortality, morbidity, and financial and societal costs, especially with the lack of novel drugs to combat infections caused by antimicrobial-resistant pathogens. Unfortunately, antimicrobial resistance (AMR) has increased significantly worldwide in the past decades, particularly in developing third-world countries. Hence, various key organisations have made AMR the focus of highly visible reports, conferences and actions. Nevertheless, the majority of investigations were initiated in the developed world. In Lebanon, as in other developing countries, AMR remains responsible for significant morbidity and mortality. Whilst a large number of studies have targeted the AMR crisis in Lebanese clinical settings, its epidemiological situation in animals, foodstuffs and the environment is still unclear in this country. In this context, and in order to better survey, control and prevent the emergence of multidrug-resistant bacterial strains in Lebanese non-clinical settings, this work reviewed the available data on the epidemiology of antimicrobial-resistant bacteria isolated from Lebanese extra-hospital sources., (Copyright © 2018. Published by Elsevier Ltd.)

Objectives: This study aimed to isolate and characterise extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) isolates from animals and wastewater in Tunisia., Methods: ESBL-E from wastewater (n=123 samples), faeces of healthy animals (poultry, sheep, goats and calves) (n=140) and raw milk from healthy cows (n=42) and goats (n=20) were investigated. Antimicrobial susceptibility was determined according to CLSI recommendations. The bla TEM , bla SHV , bla CTX-M and bla OXA-48 genes were analysed by PCR and sequencing. Phylogenetic groups were determined by PCR for Escherichia coli isolates. The clonality of E. coli and Klebsiella pneumoniae isolates was determined by XbaI-PFGE and MLST., Results: A total of 81 E. coli, 20 K. pneumoniae, 4 Enterobacter cloacae, 1 Citrobacter freundii and 1 Citrobacter braakii were isolated. The bla CTX-M-1 and bla CTX-M-15 genes were predominant in E. coli and K. pneumoniae isolates. E. cloacae and C. braakii isolates harboured the bla SHV-12 gene. The C. freundii isolated from wastewater carried bla CTX-M-15 , bla TEM-1 and bla OXA-204 . E. coli isolates belonged to phylogroups A (37), B1 (25), B2 (7) and D (12). Seventy-eight E. coli isolates were typeable by PFGE and were classified into 34 pulsotypes. The K. pneumoniae isolates belonged to 11 pulsotypes. The E. coli isolates belonged to sequence types ST131, ST224, ST162, ST845, ST5204, ST69, ST141 and ST10. The K. pneumoniae isolates belonged to ST405, ST147, ST564, ST307, ST152, ST45, ST661 and ST1564., Conclusion: This is the first report of O25b-B2 3 -CTX-M-27-ST131 E. coli isolates and of C. freundii carrying bla CTX-M-15 , bla TEM-1 and bla OXA-204 in Tunisia., (Copyright © 2019 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

The objectives of this study were to describe the frequency of antimicrobial resistance (AMR) in Escherichia coli and Campylobacter spp. isolates in fecal samples from beef cow-calf herds and to examine the associations between herd management practices, reported antimicrobial use, and AMR. Baseline prevalence data are needed to evaluate the effectiveness of antimicrobial stewardship programs. A pooled fecal sample, representing 20 cows, was collected from each of 107 herds during pregnancy testing. In the 305 recovered E. coli isolates (maximum 3 per herd), resistance to ≥ 1 antimicrobial was identified in 12 isolates [4%, 95% confidence interval (CI): 2% to 7%] from 105 herds (11%, 95% CI: 7% to 19%). The most common resistances identified in E. coli isolates were to tetracycline (3%) and to both streptomycin and sulfisoxazole (3%). Only 1 E. coli isolate was resistant to an antimicrobial of very high importance to human health - amoxicillin/clavulanic acid. However, 2 E. coli isolates had intermediate susceptibility to ciprofloxacin. Resistance to 1 antimicrobial was identified in 16 of 87 Campylobacter spp. isolates (18%, 95% CI: 11% to 28%) from 87 herds. Resistance to tetracycline was reported in 15% of Campylobacter spp. isolates and to nalidixic acid in 3.4%. Herds in which cows were treated with florfenicol were more likely to have E. coli resistance to ≥ 2 antimicrobials (OR 7.1, 95% CI: 1.1 to 57, P = 0.03). Herds with calf mortality of > 5% were more likely to have E. coli with resistance to streptomycin and sulfisoxazole [odds ratio (OR): 7.8, P = 0.03]. The results of this study are consistent with previous reports from western Canada and provide a starting point for designing an ongoing antimicrobial surveillance program.

The aim of this study was to evaluate the effects of antibiotic residues in milk on growth, ruminal fermentation, and microbial community of dairy calves in their first 35 d of age. Twenty newborn Holstein bull calves were assigned to 1 of 2 treatments equally: milk replacer without antibiotics (control) and milk replacer plus 4 antibiotics: 0.024 mg/L of penicillin, 0.025 mg/L of streptomycin, 0.1 mg/L of tetracycline, and 0.33 mg/L of ceftiofur (ANT). Starter intake and fecal consistency scores of each calf were recorded on a daily basis. Body weight, withers height, body length, and heart girth were measured on d 1, 7, 14, 21, 28, and 35 before feeding in the morning. Rumen fluid was collected on d 15, 25, and 35 to determine ruminal pH, volatile fatty acids (VFA), and NH 3 -N concentrations. A total of 10 (5 per treatment) samples of rumen fluid taken on d 35 were analyzed for microbial community. Rumen tissues from the cranial ventral sac and cranial dorsal sac were collected from 8 calves of each group for morphology analysis on d 35 after being harvested. The results showed that calves in 2 treatments had similar starter intake, body weight, withers height, body length, heart girth, and average daily gain. The ANT group showed a lower diarrhea frequency in wk 4, and no differences were found for other weeks. Calves in the ANT group exhibited a greater concentration of acetic acid in the rumen and no differences for other VFA, total VFA, rumen pH, or NH 3 -N. As for rumen morphology, the length of papillae from cranial ventral sac of the ANT group was longer than that of the control group. The results of ruminal microbial community showed that antibiotic residues had minor effects on bacteria phyla and bacteria diversity. At the genus level, calves in the ANT group showed lower richness of Prevotella and higher richness of Acetitomaculum. In conclusion, antibiotic residues stimulated the development of ruminal papillae and increased the production of acetic acid in rumen, which might be caused by the influence of antibiotics on the ruminal microbial community., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Objectives: The spread of carbapenem-resistant Enterobacteriaceae (CRE) in healthcare settings challenges clinicians worldwide. However, little is known about dissemination of CRE in livestock, food, and companion animals and potential transmission to humans., Methods: We performed a systematic review of all studies published in the PubMed database between 1980 and 2017 and included those reporting the occurrence of CRE in samples from food-producing and companion animals, wildlife, and exposed humans. The primary outcome was the occurrence of CRE in samples from these animals; secondary outcomes included the prevalence of CRE, carbapenemase types, CRE genotypes, and antimicrobial susceptibilities., Results: We identified 68 articles describing CRE among pigs, poultry, cattle, seafood, dogs, cats, horses, pet birds, swallows, wild boars, wild stork, gulls, and black kites in Africa, America, Asia, Australia, and Europe. The following carbapenemases have been detected (predominantly affecting the genera Escherichia and Klebsiella): VIM, KPC, NDM, OXA, and IMP. Two studies found that 33-67% of exposed humans on poultry farms carried carbapenemase-producing CRE closely related to isolates from the farm environment. Twenty-seven studies selectively screened samples for CRE and found a prevalence of <1% among livestock and companion animals in Europe, 2-26% in Africa, and 1-15% in Asia. Wildlife (gulls) in Australia and Europe carried CRE in 16-19%., Conclusions: The occurrence of CRE in livestock, seafood, wildlife, pets, and directly exposed humans poses a risk for public health. Prospective prevalence studies using molecular and cultural microbiological methods are needed to better define the scope and transmission of CRE., (Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Objectives: To sequence the genomes and determine the genetic mechanisms for linezolid resistance identified in three strains of Enterococcus isolated from cattle and swine caecal contents as part of the US National Antimicrobial Resistance Monitoring System (NARMS) surveillance programme., Methods: Broth microdilution was used for in vitro antimicrobial susceptibility testing to assess linezolid resistance. Resistance mechanisms and plasmid types were identified from data generated by WGS on Illumina® and PacBio® platforms. Conjugation experiments were performed to determine whether identified mechanisms were transmissible., Results: Linezolid resistance plasmids containing optrA were identified in two Enterococcus faecalis isolates and one Enterococcus faecium. The E. faecium isolate also carried the linezolid resistance gene cfr on the same plasmid as optrA. The linezolid resistance plasmids had various combinations of additional resistance genes conferring resistance to phenicols (fexA), aminoglycosides [spc and aph(3')-III] and macrolides [erm(A) and erm(B)]. One of the plasmids was confirmed to be transmissible by conjugation, resulting in linezolid resistance in the transconjugant., Conclusions: To the best of our knowledge, this is the first identification of linezolid resistance in the USA in bacteria isolated from food animals. The oxazolidinone class of antibiotics is not used in food animals in the USA, but the genes responsible for resistance were identified on plasmids with other resistance markers, indicating that there may be co-selection for these plasmids due to the use of different antimicrobials. The transmissibility of one of the plasmids demonstrated the potential for linezolid resistance to spread horizontally. Additional surveillance is necessary to determine whether similar plasmids are present in human strains of Enterococcus.

Objectives: Domestic animals can serve as a reservoir for antimicrobial-resistant Staphylococcus aureus and may lead to the emergence of drug-resistant human isolates. The present study aimed to assess the prevalence and antibiotypes of S. aureus and methicillin-resistant S. aureus (MRSA) cultured from clinical pyogenic cases of domestic animals in India., Methods: A total of 100 samples were collected during the study (August 2014 to July 2015) from different pyogenic conditions of cattle (n=21), buffaloes (n=63) and dogs (n=16). Samples were processed for isolation and phenotypic identification of S. aureus. Species identification was confirmed by PCR, and all of the isolates were screened for the mecA gene. Antimicrobial susceptibility testing was performed by the disk diffusion method for 14 antimicrobial agents., Results and Conclusion: The study revealed a 40.0% prevalence of S. aureus in pyogenic clinical cases of domestic animals. The species-wise prevalence of S. aureus was 38.1% both in cattle and buffaloes and 50.0% in dogs. The study revealed 21 antibiotypes among the S. aureus isolates. The rate of resistance was highest for amoxicillin (95.0%), followed by penicillin G (82.5%). Among the 40 S. aureus isolates, 23 (57.5%) exhibited methicillin resistance and carried the mecA gene. To the best of our knowledge based on the available literature, this is the first report of its kind from this country., (Copyright © 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Antimicrobial resistance is a complex issue with a large volume of published literature, and there is a need for synthesis of primary studies for an integrated understanding of this topic. Our research team aimed to have a more complete understanding of antimicrobial resistance in Canada (IAM.AMR Project) using multiple methods including the literature reviews and quantitative modelling. To accomplish this goal, qualitative features of publications (e.g., geographical location, study population) describing potential relationships between the occurrence of antimicrobial resistance and factors (e.g., antimicrobial use; management system) were of particular interest. The objectives of this review were to (a) describe the available peer-reviewed literature reporting potential relationships between factors and antimicrobial resistance; and (b) to highlight data gaps. A comprehensive literature search and screening were performed to identify studies investigating factors potentially linked with antimicrobial resistance in Campylobacter species, Escherichia coli and Salmonella enterica along the farm-to-fork pathway (farm, abattoir (slaughter houses) and retail meats) for the major Canadian livestock species (beef cattle, broiler chicken and pigs). The literature search returned 14,966 potentially relevant titles and abstracts. Following screening of titles, abstracts and full-text articles, the qualitative features of retained studies (n = 28) were extracted. The most common factors identified were antimicrobial use (n = 13 studies) and type of farm management system (e.g., antibiotic-free, organic; n = 8). Most studies were conducted outside of Canada and involved investigations at the farm level. Identified data gaps included the effect of vaccination, industry-specific factors (e.g., livestock density) and factors at sites other than farm along the agri-food chain. Further investigation of these factors and other relevant industry activities are needed for the development of quantitative models that aim to identify effective interventions to mitigate the occurrence of antimicrobial resistance along the agri-food chain., (© 2018 Blackwell Verlag GmbH.)

To assess phenotypic bacterial antimicrobial resistance (AMR) in different strata (e.g., host populations, environmental areas, manure, or sewage effluents) for epidemiological purposes, isolates of target bacteria can be obtained from a stratum using various sample types. Also, different sample processing methods can be applied. The MIC of each target antimicrobial drug for each isolate is measured. Statistical equivalence testing of the MIC data for the isolates allows evaluation of whether different sample types or sample processing methods yield equivalent estimates of the bacterial antimicrobial susceptibility in the stratum. We demonstrate this approach on the antimicrobial susceptibility estimates for (i) nontyphoidal Salmonella spp. from ground or trimmed meat versus cecal content samples of cattle in processing plants in 2013-2014 and (ii) nontyphoidal Salmonella spp. from urine, fecal, and blood human samples in 2015 (U.S. National Antimicrobial Resistance Monitoring System data). We found that the sample types for cattle yielded nonequivalent susceptibility estimates for several antimicrobial drug classes and thus may gauge distinct subpopulations of salmonellae. The quinolone and fluoroquinolone susceptibility estimates for nontyphoidal salmonellae from human blood are nonequivalent to those from urine or feces, conjecturally due to the fluoroquinolone (ciprofloxacin) use to treat infections caused by nontyphoidal salmonellae. We also demonstrate statistical equivalence testing for comparing sample processing methods for fecal samples (culturing one versus multiple aliquots per sample) to assess AMR in fecal Escherichia coli These methods yield equivalent results, except for tetracyclines. Importantly, statistical equivalence testing provides the MIC difference at which the data from two sample types or sample processing methods differ statistically. Data users (e.g., microbiologists and epidemiologists) may then interpret practical relevance of the difference. IMPORTANCE Bacterial antimicrobial resistance (AMR) needs to be assessed in different populations or strata for the purposes of surveillance and determination of the efficacy of interventions to halt AMR dissemination. To assess phenotypic antimicrobial susceptibility, isolates of target bacteria can be obtained from a stratum using different sample types or employing different sample processing methods in the laboratory. The MIC of each target antimicrobial drug for each of the isolates is measured, yielding the MIC distribution across the isolates from each sample type or sample processing method. We describe statistical equivalence testing for the MIC data for evaluating whether two sample types or sample processing methods yield equivalent estimates of the bacterial phenotypic antimicrobial susceptibility in the stratum. This includes estimating the MIC difference at which the data from the two approaches differ statistically. Data users (e.g., microbiologists, epidemiologists, and public health professionals) can then interpret whether that present difference is practically relevant., (Copyright © 2018 Shakeri et al.)

The antibiotic monensin is fed to dairy cows to increase milk production efficiency. A fraction of this monensin is excreted into the cow manure. Previous studies have found that cow manure containing monensin can negatively impact the performance of anaerobic digesters, especially upon first introduction. Few studies have examined whether the anaerobic digester microbiome can adapt to monensin during the operating time. Here, we conducted a long-term time series study of four lab-scale anaerobic digesters fed with cow manure. We examined changes in both the microbiome composition and function of the anaerobic digesters when subjected to the dairy antibiotic monensin. In our digesters, monensin was not rapidly degraded under anaerobic conditions. The two anaerobic digesters that were subjected to manure from monensin feed-dosed cows exhibited relatively small changes in microbiome composition and function due to relatively low monensin concentrations. At higher concentrations of monensin, which we dosed directly to control manure (from dairy cows without monensin), we observed major changes in the microbiome composition and function of two anaerobic digesters. A rapid introduction of monensin to one of these anaerobic digesters led to the impairment of methane production. Conversely, more gradual additions of the same concentrations of monensin to the other anaerobic digester led to the adaptation of the anaerobic digester microbiomes to the relatively high monensin concentrations. A member of the candidate OP11 ( Microgenomates ) phylum arose in this anaerobic digester and appeared to be redundant with certain Bacteroidetes phylum members, which previously were dominating. IMPORTANCE Monensin is a common antibiotic given to dairy cows in the United States and is partly excreted with dairy manure. An improved understanding of how monensin affects the anaerobic digester microbiome composition and function is important to prevent process failure for farm-based anaerobic digesters. This time series study demonstrates how anaerobic digester microbiomes are inert to low monensin concentrations and can adapt to relatively high monensin concentrations by redundancy in an already existing population. Therefore, our work provides further insight into the importance of microbiome redundancy in maintaining the stability of anaerobic digesters., (Copyright © 2018 Spirito et al.)

The aim of this study was to describe and compare the occurrence of quinolone resistant Escherichia coli (QREC) in various animal species in relation to human population density. Data from the Norwegian monitoring programme for antimicrobial resistance in feed, food and animals from 2006 to 2016 was compiled and analysed. In total, 4568 E. coli isolates were included in this study. The isolates originated from broilers, layers, cattle, turkeys, dogs, wild birds, red foxes, reindeer, sheep, horses and pigs. Data regarding the geographical location of sampling was obtained for 4050 of these isolates and used to categorize the isolates depending on the human population density of the area. In total, 1.4% of the isolates were categorized as quinolone resistant. Compared to most European countries, there was an overall low occurrence of QREC in various animal species in Norway, though with an interspecies variation with the highest occurrence in broilers and wild birds (p < 0.05). Human population density was not associated with the occurrence of QREC. Since fluoroquinolones are not used prophylactically and in almost negligent amounts in various species in Norway, the interspecies variation in the occurrence of QREC suggests that other factors than fluoroquinolone use may be important in the development of QREC., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

In this retrospective study, we describe and analyse Salmonella data from four livestock species in Great Britain between 1983 and 2014, focusing on Salmonella Typhimurium. A total of 96 044 Salmonella isolates were obtained during the study period. S. Typhimurium was the predominant serovar isolated from cattle and pigs and represented 40.7% (18 455/45 336) and 58.3% (4495/7709) of isolates from these species respectively, while it only accounted for 6.7% (2114/31 492) of chicken isolates and 8.1% (926/11 507) of turkey isolates. Over the study period, DT104 was the most common phage type in all four species; however, DT104 peaked in occurrence between 1995 and 1999, but is currently rare. Monophasic strains of S. Typhimurium represented less than 3% of all Salmonella isolates in cattle and chickens in 2014, but accounted for 10.4% of all turkey isolates and 39.0% of all pig isolates in the same year. Salmonella isolates were tested for their in vitro susceptibility to 16 antimicrobials. Antimicrobial resistance of S. Typhimurium isolates is largely influenced by the dominance of specific phage types at a certain time, which are commonly associated with particular resistance patterns. Changes in resistance patterns over time were analysed and compared between species.

The European Antimicrobial Susceptibility Surveillance in Animals (EASSA) program collects zoonotic and commensal bacteria from food-producing animals at slaughter and tracks their susceptibility to medically important antibiotics. Results of commensal enterococci species (2013-2014) are presented here. Intestinal content from cattle, pigs and chickens were randomly sampled (5-6 countries/host; ≥4 abattoirs/country; 1 sample/animal/farm) for isolation of enterococci, MICs of 9 antibiotics were assessed by CLSI agar dilution in a central laboratory. Clinical breakpoints (CLSI) and epidemiological cut-off values (EUCAST) were applied for data interpretation. In total 960 Enterococcus faecium and 779 Enterococcus faecalis strains were recovered. Seven porcine E. faecium/faecalis strains of Spanish origin were resistant to linezolid. One avian E. faecalis and one porcine E. faecium strain were non-wild type (MICs 8 mg/L) to daptomycin. Clinical vancomycin resistance was absent; 2 poultry E. faecium and 1 bovine E. faecalis strains were non-wild type, all with MICs of 8 mg/L. None of the strains tested were clinically resistant to tigecycline. Little clinical resistance to ampicillin or gentamicin was observed. Clinical resistance of E. faecium to quinupristin/dalfopristin was slightly higher (2.2-12.0%) but 61.9-83.2% of the strains were classified as non-wild type. Very high percentages resistance to tetracycline (67.4-78.3%) and to erythromycin (27.1-57.0%) were noted for both E. faecium and E. faecalis in pigs and chickens compared to cattle (5.2-30.4 and 9.0-10.4%, respectively). Similar non-wild type results were observed for E. hirae (n = 557), E. durans (n = 218) and E. casseliflavus (n = 55) including percentage non-wild type for daptomycin, linezolid, tigecycline being absent and for vancomycin low. For these species percentage non-wild type to erythromycin was lower as compared to E. faecalis/faecium. This pan-EU survey shows high variability in antibiotic susceptibility of commensal enterococci from healthy food animals. Clinical resistance to critically important antibiotics for human medicine was absent or low, except for erythromycin., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The aim of this study is to investigate the biopotency of methanolic extracts of Vitex mollis, Psidium guajava, Dalbergia retusa, and Crescential alata leaves against various staphylococcal strains isolated from cattle and rabbits. Methicillin-resistant S. aureus strains were isolated from cattle, while other strains were isolated from rabbits using standard methodology. The total phytochemical phenolic and saponins contents were obtained being the main groups of the antinutritional factors. The antimicrobial activity of the extracts against the standard culture of S. aureus (control) and S. aureus isolated from cattle and rabbits were investigated comparatively relative to that of oxacillin. It was found that both the control S. aureus and the isolated S. aureus are susceptible to all the four plant extracts, and sensitive to oxacillin. Of all the S. aureus including the control, MRSA2 is the most susceptible to all the extracts at 1000 μg/mL, except that of V. mollis where it is the least susceptible. Among all the plant extracts, P. guajava is the most active against MRSA2 and SOSA2. Therefore, the isolates from cattle (MRSA1 and MRSA2) are more susceptible to all the plant extracts than the isolates from rabbits. Among all the rabbit isolates, CoNS3 is the least susceptible to the extracts. Since all the plant extracts exhibit remarkable inhibitory activities against all the S. aureus strains, they are promising towards the production of therapeutic drugs., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The desideratum aim of the present context was to assess the biopotency of methanolic extracts of Eichhornia crassipes (E. crassipes), Pistacia vera (P. vera), and Ziziphus amole (Z. amole) leaves against various staphylococcal strains, and to quantify the phenolics as well as saponin content in them. The antibacterial activity of various concentrations (62.5-1000 μg/mL) of plant extracts was tested against control clinical strains (Staphylococcus aureus ATCC 25923, S. aureus ATCC 29213, and S. aureus ATCC 43300), methicillin-resistant S. aureus (MRSA1 and MRSA2), oxacillin sensitive S. aureus (SOSA1 and SOSA2), and coagulase-negative Staphylococcus epidermidis (CoNS1, CoNS2, and CoNS3) using disc diffusion assay. Leaf extracts of the three plants exhibited pronounced growth inhibitory characteristics against staphylococci in a dose dependent manner. E. crassipes extract depicted the highest relative percentage inhibition values against control clinical strains (68.6 ± 0.5%), while P. vera (68.6 ± 0.3%) and Z. amole (74.79 ± 0.3%) extracts showed pronounced relative inhibition values against staphylococcal strains isolated from cattle. Total phenols and saponin content of leaf extracts were investigated by standard in vitro methods. The methanolic extracts of these plants were found to comprise substantial content of phenolics and saponin at varying levels. The highest value of phenolics was estimated in P. vera extract (60.0 ± 1.3 mg gallic acid/g extract), followed by Z. amole (33.6 ± 1.4 mg gallic acid/g extract), and E. crassipes (23.0 ± 1.3 mg gallic acid/g extract). Saponin content for P. vera, Z. amole, and E. crassipes extracts were estimated as 41.0 ± 1.3, 35.8 ± 1.3, and 25.0 ± 1.2 mg diosgenin/g extract, respectively. The outcome of this study suggested the exploitation of methanolic extract of P. vera, Z. amole, and E. crassipes leaves for their possible application in ethnomedicine, particularly as drugs preparation against staphylococcal infections. In conclusion, the study indicates the biopotency of these plants against pathogenic MRSA present in cattle, and SOSA as well as CoNS bacteria present in rabbits, which could be a serious issue for livestock., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Escherichia coli (E. coli) is one of the most relevant opportunistic pathogenic bacteria as it may cause severe morbidity and mortality in yaks (poephagus grunniens). In recent years, several kinds of antibiotics have been widely used in Tibetan areas to treat the bacterial diseases, resulting in serious repercussions on the bacterial antibiotic resistance in yaks. This investigation was conducted in order to determine the prevalence of antimicrobial resistance and integron gene cassettes in E. coli isolated from yaks in Aba Tibetan Autonomous Prefecture (Aba TAP), China., Methods: A total of 278 non-duplicated fresh samples were collected from the yaks in Aba TAP for the isolation and identification of E. coli isolates. Antimicrobial susceptibility testing is performed by using the disc diffusion method according to the Clinical and Laboratory Standards Institute guidelines (CLSI, 2013). Various antibiotic resistance genes and integron gene cassettes were detected by polymerase chain reaction (PCR) and sequencing., Results: Overall, a total of 228 E. coli bacteria were isolated from the fresh faeces of yaks in four different geographical regions. 58% of those isolates showed multi-drug resistance capabilities (MDR) in our study. These isolated bacteria showed a high resistance rate to streptomycin (84%), cefotaxime (79%), amikacin (61%) and trimethoprim-sulfamethoxazole (54%). The most common antimicrobial resistance genes in the isolates were bla CTX-M , sul1, aph (3')-IIa, aac (3)-IIa, aac (6')-Ib, tetB, with respective detection rates of 65%, 46%, 35%, 13%, 11%, and 10%. Furthermore, 66% and 6% of the strains carried Class 1 and 2 integrons, respectively. However, the class 3 integron was not detected. Gene cassette arrays in the class 1 integron included aadA1, aadA7, aadA5, aadA17, dfrA1, dfrA5, dfrA1-aadA1, dfrA12-aadA2 and dfrA17-aadA5. The most prevalent gene cassette was aadA1 (20%). For the class 2 integron, dfrA1-sat2-aadA1 (6%) and dfrA1-sat1-aadA1 (0.4%) were also detected as part of this research., Conclusion: High multi-drug resistance rates have been discovered, as well as a prevalence of antibiotic resistance genes and integron gene cassettes in the E. coli isolated from the faeces of yak. This might create a potential problem for treatment of the yaks' bacterial infections as well as food hygiene for humans. It is therefore urgently necessary to begin continuous surveillance and analysis of antibiotic resistance and integron cassettes in other bacteria from yaks., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Ionophores and antibiotics have been shown to decrease ruminal methanogenesis both in vitro and in vivo but have shown little evidence toward a sustainable means of mitigation. Feed additive rotation was proposed and investigated for methane, VFA, and microbial population response. In the present study, cannulated steers ( = 12) were fed a moderate-forage basal diet in a Calan gate facility for 13 wk. In addition to the basal diet, steers were randomly assigned to 1 of 6 treatments: 1) control, no additive; 2) bambermycin, 20 mg bambermycin/d; 3) monensin, 200 mg monensin/d; 4) the basal diet + weekly rotation of bambermycin and monensin treatments (B7M); 5) the basal diet + rotation of bambermycin and monensin treatments every 14 d (B14M); and 6) the basal diet + rotation of bambermycin and monensin treatments every 21 d (B21M). Steers were blocked by weight in a randomized complete block design where the week was the repeated measure. Rumen fluid was collected weekly for analysis ( = 13), and results were normalized according to individual OM intake (OMI; kg/d). Potential activity of methane production was not significantly different among treatments ( > 0.05). However, treatment tended to affect the CH-to-propionate ratio ( = 0.0565), which was highest in the control and lowest in the monensin, B21M, and B14M treatments (0.42 vs. 0.36, 0.36, and 0.33, respectively). The CH:propionate ratio was lowest in wk 2 and 3 ( < 0.05) but the ratio in wk 4 to 12 was not different from the ratio in wk 0. Week also affected total VFA, with total VFA peaking at wk 3 and plummeting at wk 4 (4.02 vs. 2.86 m/kg OMI; < 0.05). A significant treatment × week interaction was observed for the acetate-to-propionate (A:P) ratio, where bambermycin- and rotationally fed steers did not have a reduced A:P ratio compared with monensin-fed steers throughout the feeding period ( < 0.0001). Microbial analysis revealed significant shifts, but several predominant classes showed adaptation between 4 and 6 wk after additive initiation. There was no significant evidence to suggest that rotations of monensin and bambermycin provided additional benefits to steers consuming a moderate-forage diet at the microbial/animal and environmental level versus those continuously fed.

Several factors are known to affect the intestinal microbiota of cattle. However, how these changes occur over time is poorly understood. This study aimed to investigate the consequences of entrance into a feedlot and the effects of virginiamycin used as a growth promoter on the bovine fecal microbiota. Two batches of beef cattle (B1, n=50 and B2, n=36) entering a feedlot operation were randomly divided into two pens: one receiving virginiamycin and one group not receiving antibiotic (control group). Fecal samples were collected at arrival, mid feedlot and at exit to slaughter. The V4 region of 16S rRNA gene was amplified and sequenced. Escherichia coli strains isolated in samples from arrival and exit of B2 were also isolated and used as indicators of antimicrobial susceptibility. Marked changes in membership and structure of fecal microbiota occurred following entrance into the feedlot. At mid feedlot, virginiamycin affected bacterial community membership, but not structure, suggesting that the antibiotic had a stronger effect on the rare, but not on the most abundant species. The use of virginiamycin had no demonstrable effect on antibiotic resistance in E. coli. The differences found between batches are suggestive that variations in study conditions are important and can strongly affect the intestinal microbiota., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Aim: We report a one-health investigation of three cases of leptospirosis on a dairy farm with unvaccinated cattle in New Zealand. The cases are discussed in the context of diagnostics, risk factors, persistence of symptoms and outbreak mitigation measures., Method: Clinical and laboratory records from the human cases were reviewed and serological and molecular investigations were conducted into the Leptospira status of cattle and pigs on the farm., Results: Cases presented early in their illness and all three were confirmed within seven days of onset of symptoms by urine PCR and within 18 days by convalescent MAT (two Hardjo, one Pomona). Cattle and pigs had serological evidence of recent infection with Hardjo/Pomona and Pomona/Copenhageni respectively. Pigs were slaughtered and cattle were vaccinated. Post-exposure prophylaxis was given to staff in-contact with the milking herd until the herd had antibiotic treatment at drying-off (approximately four months after the initial case)., Conclusion: The utility of PCR testing for Leptospira DNA as both an early and rapid test for leptospirosis was demonstrated. Two of three cases reported persistence of symptoms at least six months after the acute episode and one of these remains unable to work. Risk mitigation measures such as post-exposure prophylaxis, animal vaccination, heightened clinical suspicion of leptospirosis and recognition of context specific risk factors (eg, effluent spreading) demonstrate the value of medical and veterinary experts working together., Competing Interests: Nil.

To enable future comparison of the antimicrobial susceptibility data between bacteria obtained from animals and humans, it is necessary to compare the relationships between minimum inhibitory concentrations (MICs) of veterinary and human medicine. We evaluated the relationship between the MIC of ceftiofur (CTF) and the MICs of other third-generation cephalosporins (TGCs): cefotaxime (CTX), cefpodoxime (CPDX), and ceftazidime (CAZ), determined by the broth microdilution method using 118 cefazolin-resistant Escherichia coli isolates from food-producing animals. Using the Clinical and Laboratory Standards Institute criteria, very major classification errors were observed only in CAZ (17.8%, 21 of 118); major and minor errors were observed in all TGCs (CTX: 0.8% [1 of 118] and 9.3% [11 of 118]; CPDX: 9.3% [11 of 118] and 6.8% [8 of 118]; CAZ: 2.5% [3 of 118] and 9.3% [11 of 118], respectively). The Spearman correlation coefficients between the MICs of CTF and CTX, CPDX, and CAZ were 0.765, 0.731, and 0.306, respectively. The sensitivity and specificity values were 100.0% and 81.8% for CTX, 99.0% and 27.3% for CPDX, and 76.0% and 86.4% for CAZ compared with CTF. The C-statistic was 0.978 for CTF and CTX, 0.953 for CPDX, and 0.798 for CAZ. For the TGCs evaluated in our study, testing for CTX susceptibility results showed the highest correlation with the results given when testing for CTF susceptibility.

Colistin resistance genes mcr-3 and mcr-1 have been detected in an Escherichia coli isolate from cattle faeces in a Spanish slaughterhouse in 2015. The sequences of both genes hybridised to same plasmid band of ca 250 kb, although colistin resistance was non-mobilisable. The isolate was producing extended-spectrum beta-lactamases and belonged to serotype O9:H10 and sequence type ST533. Here we report an mcr-3 gene detected in Europe following earlier reports from Asia and the United States., (This article is copyright of The Authors, 2017.)

We investigated the occurrence of integrons in antibiotic-resistant Escherichia coli strains isolated from free-ranging food animals, including yaks, piglets, and chickens, in China, and characterized the gene cassettes harbored within the integrons. We examined 432 E. coli strains that exhibited resistance to at least one class of antibiotics. Integrase genes and associated gene cassettes were characterized by polymerase chain reaction (PCR) analysis, restriction fragment-length polymorphism, DNA sequencing, conjugation experiments, and plasmid analysis. Twenty-nine (6.7%) integrons were amplified from the 432 antimicrobial-resistant (AMR) isolates evaluated. Specifically, class 1 and 2 integrons were detected in 26 (6%) and 3 (0.7%) strains, respectively. Meanwhile, 6 different gene cassettes, dfrA1, dfr12, aadA1, aadA2, sat1, and orfF, were detected within 6 variable regions (VRs), of which the dfrA1 + aadA1 array was the most common, identified in 12 of 26 class 1 integrons (46.1%). Meanwhile, only one class 2 integron contained a cassette, and the remaining two contained undetermined VRs. Finally, a conjugation assay confirmed the transfer of 4 different types of class 1 integrons into recipient strains, with plasmid sizes ranging from 20 to 30 kb. This is the first report examining the baseline AMR characteristics of E. coli within an extensive farming system of livestock animals in China. Given that integrons were detected in >6% of resistant E. coli strains, precautionary measures are required to prevent the spread of mobile genetic resistance determinants in food animals and monitor their emergence., (© 2017 Institute of Food Technologists®.)

Antimicrobial agents are used in cattle production systems for the prevention and control of bacterial associated diseases. A consequence of their use is the potential development of antimicrobial resistance (AMR). Enterococcus faecium and Enterococcus faecalis that are resistant to antimicrobials are of increased concern to public health officials throughout the world as they may compromise the ability of various treatment regimens to control disease and infection in human medicine. Australia is a major exporter of beef; however it does not have an ongoing surveillance system for AMR in cattle or foods derived from these animals. This study examined 910 beef cattle, 290 dairy cattle and 300 veal calf faecal samples collected at slaughter for the presence of enterococci. Enterococcus were isolated from 805 (88.5%) beef cattle faeces, 244 (84.1%) dairy cattle faeces and 247 (82.3%) veal calf faeces with a total of 800 enterococci subsequently selected for AMR testing. The results of AMR testing identified high levels of resistance to antimicrobials that are not critically or highly important to human medicine with resistance to flavomycin (80.2%) and lincomycin (85.4-94.2%) routinely observed. Conversely, resistance to antibiotics considered critically or highly important to human medicine such as tigecycline, daptomycin, vancomycin and linezolid was not present in this study. There is minimal evidence that Australian cattle production practices are responsible for disproportionate contributions to AMR development and in general resistance to antimicrobials of critical and high importance in human medicine was low regardless of the isolate source. The low level of antimicrobial resistance in Enterococcus from Australian cattle is likely to result from comprehensive controls around the use of antimicrobials in food-production animals in Australia. Nevertheless, continued monitoring of the effects of all antimicrobial use is required to support Australia's reputation as a supplier of safe and healthy food.

The prevalence and antibiotic resistance of Acinetobacter spp. from fifty samples of meat (chicken, turkey, beef and pork) were evaluated. Acinetobacter spp. was recovered from all samples and the clonal relatedness of 223 isolates identified to belong to the genus Acinetobacter was established by PFGE. A high genetic diversity was observed and 166 isolates from different samples, 141 representing different PFGE profiles, were further identified to the species level by rpoB gene sequencing. Thirteen distinct Acinetobacter species were identified among 156 isolates. The remaining ten isolates may represent three putatively novel species since rpoB sequence homologies with type strains of all available described Acinetobacter species, were <95%. The most common species was Acinetobacter guillouiae with a prevalence of 34.9%. However 18.7% of the strains belong to the Acinetobacter baumannii group (n=31) which include the species Acinetobacter baumannii (n=7), Acinetobacter pittii (n=12), Acinetobacter seifertii (n=8) and Acinetobacter nosocomialis (n=4) that are the species most frequently associated with nosocomial infections worldwide. In general, strains were resistant to some of the antimicrobials most frequently used to treat Acinetobacter infections such as piperacillin-tazobactam (64.9% of strains resistant), ceftazidime (43.5%), ciprofloxacin (42.9%), as well as to colistin (41.7%) and polymyxin B (35.1%), the last-resort drugs to treat infections caused by multidrug-resistant Acinetobacter. The percentage of resistant strains to trimethoprim-sulfamethoxazole, tetracycline, aminoglycosides (amikacin and tobramycin) and ampicillin-sulbactam was >10% (23.2%, 23.2%, 14.3%, 12.5%, 12.5%, respectively). However, resistances to meropenem, imipenem and minocycline were only sporadically observed (8.3%, 1.2% and 1.2%, respectively). Overall, 51.2% of the strains were considered as multidrug-resistant (MDR) and 9.6% as extensively drug-resistant (XDR). The prevalence of MDR strains within the A. baumannii group (38.7%) was lower than the prevalence within the others species identified (54.1%). Therefore, food of animal origin may be a vehicle of spread Acinetobacter strains resistant to several antibiotics in the community and in the hospital setting environment. This may led to nosocomial and community-acquired infections in susceptible individuals., (Copyright © 2016 Elsevier B.V. All rights reserved.)