Antimicrobial Resistance and Virulence of Enterococcus faecalis Isolated from Retail Food†

Although enterococci are considered opportunistic nosocomial pathogens, their contribution to foodborne illnesses via dissemination through retail food remains undefined. In this study, prevalence and association of antimicrobial resistance and virulence factors of 80 Enterococcus faecalis isolates from retail food items were investigated. The highest rates of resistance were observed for lincomycin (73 of 80 isolates, 91%) and bacitracin (57 of 80 isolates, 71%), and lower rates of resistance (≤40%) were found for chloramphenicol, ciprofloxacin, erythromycin, flavomycin, gentamicin, kanamycin, nitrofurantoin, penicillin, and tylosin. Overall resistance to antimicrobials was low for most isolates tested. Of the virulence factors tested, the majority of isolates were positive for ccf (78 of 80 isolates, 98%), efaAfs (77 of 80, 96%), and cpd (74 of 80, 93%). Isolates also commonly contained cob (72 of 80 isolates, 90%) and gelE (68 of 80, 85%). Very few isolates contained cylMBA (12 of 80 isolates [15%] for cylM and 9 of 80 isolates [11%] for both cylB and cylA) and efaAfm (2 of 80 isolates, 3%). Positive statistical associations (significance level of 0.05) were found between agg and tetracycline resistance, cylM and erythromycin resistance, and gelE and efaAfs and lincomycin resistance. The presence of the cylB and cylA alleles also was positively correlated with bacitracin and tetracycline resistance. Negative correlations were observed between many of the virulence attributes and resistance to ciprofloxacin, erythromycin, flavomycin, gentamicin, kanamycin, and tylosin. These data suggest that both positive and negative associations exist between antimicrobial resistance genes and virulence factors in E. faecalis isolates from foods commonly purchased from grocery stores.

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Genomic analysis of multidrug-resistant clinical Enterococcus faecalis isolates for antimicrobial resistance genes and virulence factors from the western region of Saudi Arabia

Antimicrobial Resistance and Infection Control ◽

10.1186/s13756-019-0508-4 ◽

2019 ◽

Vol 8 (1) ◽

Cited By ~ 9

Author(s):

Muhammad Farman ◽

Muhammad Yasir ◽

Rashad R. Al-Hindi ◽

Suha A. Farraj ◽

Asif A. Jiman-Fatani ◽

...

Keyword(s):

Saudi Arabia ◽

Antimicrobial Resistance ◽

Enterococcus Faecalis ◽

Virulence Factors ◽

Resistance Genes ◽

Genomic Analysis ◽

Multidrug Resistant ◽

Western Region ◽

Antimicrobial Resistance Genes ◽

The Western Region

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Characteristics of High-Level Aminoglycoside-Resistant Enterococcus faecalis Isolated from Bulk Tank Milk in Korea

Animals ◽

10.3390/ani11061724 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1724

Author(s):

Hyo Jung Kang ◽

Sunghyun Yoon ◽

Koeun Kim ◽

Young Ju Lee

Keyword(s):

Antimicrobial Resistance ◽

Enterococcus Faecalis ◽

Virulence Factors ◽

Virulence Genes ◽

Multidrug Resistant ◽

Management Program ◽

Bulk Tank Milk ◽

Antimicrobial Resistance Genes ◽

Bulk Tank ◽

High Level

Enterococci, which are considered environmental mastitis-causing pathogens, have easily acquired aminoglycoside-resistant genes that encode various aminoglycoside-modifying enzymes (AME). Therefore, this study was conducted to compare the distribution of high-level aminoglycoside-resistant (HLAR) and multidrug-resistant (MDR) Enterococcus faecalis (E. faecalis) bacteria isolated from bulk tank milk in four dairy companies in Korea. Moreover, it analyzed the characteristics of their antimicrobial resistance genes and virulence factors. Among the 301 E. faecalis bacteria studied, 185 (61.5%) showed HLAR with no significant differences among the dairy companies. Furthermore, 129 (69.7%) of the 185 HLAR E. faecalis showed MDR without significant differences among companies. In contrast, HLAR E. faecalis from companies A, B, and C were significantly higher in resistance to the four classes than those in company D, which had the highest MDR ability against the three antimicrobial classes (p < 0.05). In addition, in the distribution of AME genes, 72 (38.9%) and 36 (19.5%) of the isolates carried both aac(6′)Ie-aph(2″)-la and ant(6)-Ia genes, and the ant (6)-Ia gene alone, respectively, with significant differences among the companies (p < 0.05). In the distribution of virulence genes, the ace (99.5%), efa A (98.9%), and cad 1 (98.4%) genes were significantly prevalent (p < 0.05). Thus, our results support that an advanced management program by companies is required to minimize the dissemination of antimicrobial resistance and virulence factors.

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Diversity and Antimicrobial Resistance in the Streptococcus bovis/Streptococcus equinus Complex (SBSEC) Isolated from Korean Domestic Ruminants

Microorganisms ◽

10.3390/microorganisms9010098 ◽

2021 ◽

Vol 9 (1) ◽

pp. 98

Author(s):

Seon Young Park ◽

Mingyung Lee ◽

Se Ra Lim ◽

Hyemin Kwon ◽

Ye Seul Lee ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Phylogenetic Analyses ◽

Tetracycline Resistance ◽

Current Status ◽

Streptococcus Bovis ◽

Limited Information ◽

Phenotypic Resistance ◽

Domestic Ruminants ◽

Antimicrobial Resistance Genes ◽

Acid Producing Bacteria

S. bovis/S. equinus complex (SBSEC) includes lactic acid-producing bacteria considered as the causative agent associated with acute rumen lactic acidosis in intensive ruminants. Considering the limited information on the detailed characteristics and diversity of SBSEC in Korea and the emergence of antimicrobial resistance (AMR), we investigated the diversity of SBSEC from domestic ruminants and verified the presence of antimicrobial resistance genes (ARGs) against several antimicrobials with their phenotypic resistance. Among 51 SBSEC isolates collected, two SBSEC members (S. equinus and S. lutetiensis) were identified; sodA-based phylogenetic analyses and comparisons of overall genome relatedness revealed potential plasticity and diversity. The AMR rates of these SBSEC against erythromycin, clindamycin, and tetracycline were relatively lower than those of other SBSEC isolates of a clinical origin. An investigation of the ARGs against those antimicrobials indicated that tetracycline resistance of SBSECs generally correlated with the presence of tet(M)-possessing Tn916-like transposon. However, no correlation between the presence of ARGs and phenotypic resistance to erythromycin and clindamycin was observed. Although a limited number of animals and their SBSEC isolates were examined, this study provides insights into the potential intraspecies biodiversity of ruminant-origin SBSEC and the current status on antimicrobial resistance of the bacteria in the Korean livestock industry.

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Metagenomic Quantification of Genes with Internal Standards

mBio ◽

10.1128/mbio.03173-20 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Emily Crossette ◽

Jordan Gumm ◽

Kathryn Langenfeld ◽

Lutgarde Raskin ◽

Melissa Duhaime ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Tetracycline Resistance ◽

Dairy Manure ◽

Molecular Techniques ◽

Gene Copy ◽

Gene Quantification ◽

Gene Copies ◽

Antimicrobial Resistance Genes ◽

Metagenomic Approach

ABSTRACT We demonstrate that an assembly-independent and spike-in facilitated metagenomic quantification approach can be used to screen and quantify over 2,000 genes simultaneously, while delivering absolute gene concentrations comparable to those for quantitative PCR (qPCR). DNA extracted from dairy manure slurry, digestate, and compost was spiked with genomic DNA from a marine bacterium and sequenced using the Illumina HiSeq4000. We compared gene copy concentrations, in gene copies per mass of sample, of five antimicrobial resistance genes (ARGs) generated with (i) our quantitative metagenomic approach, (ii) targeted qPCR, and (iii) a hybrid quantification approach involving metagenomics and qPCR-based 16S rRNA gene quantification. Although qPCR achieved lower quantification limits, the metagenomic method avoided biases caused by primer specificity inherent to qPCR-based methods and was able to detect orders of magnitude more genes than is possible with qPCR assays. We used the approach to simultaneously quantify ARGs in the Comprehensive Antimicrobial Resistance Database (CARD). We observed that the total abundance of tetracycline resistance genes was consistent across different stages of manure treatment on three farms, but different samples were dominated by different tetracycline resistance gene families. IMPORTANCE qPCR and metagenomics are central molecular techniques that have offered insights into biological processes for decades, from monitoring spatial and temporal gene dynamics to tracking ARGs or pathogens. Still needed is a tool that can quantify thousands of relevant genes in a sample as gene copies per sample mass or volume. We compare a quantitative metagenomic approach with traditional qPCR approaches in the quantification of ARG targets in dairy manure samples. By leveraging the benefits of nontargeted community genomics, we demonstrate high-throughput absolute gene quantification of all known ARG sequences in environmental samples.

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Antimicrobial resistance of Enterococcus faecalis isolated from meat

Bulletin of the Veterinary Institute in Pulawy ◽

10.1515/bvip-2015-0034 ◽

2015 ◽

Vol 59 (2) ◽

pp. 229-233 ◽

Cited By ~ 5

Author(s):

Hanna Różańska ◽

Aleksandra Lewtak-Piłat ◽

Jacek Osek

Keyword(s):

Minimum Inhibitory Concentration ◽

Antimicrobial Resistance ◽

Enterococcus Faecalis ◽

Inhibitory Concentration ◽

Poultry Meat ◽

Erythromycin Resistance ◽

Concentration Technique

Abstract The aim of the study was the evaluation of the antimicrobial resistance of Enterococcus faecalis strains isolated from cattle, pig, and poultry meat. A test was performed on 111 strains using the minimum inhibitory concentration technique. The highest number of isolates (94 strains) were resistant to lincomycin, the second-highest resistance was to quinupristin/dalfopristin (88 strains), tetracycline followed (65 strains), and erythromycin resistance was also notable (40 strains). All isolates tested were sensitive to daptomycin, nitrofurantoine, and tigecycline, whereas only few strains were resistant to ciprofloxacin, gentamicin, penicillin, and vancomycin. The obtained results showed that meat may be a source of antimicrobial resistant enterococci which may be transferred to humans

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Multiple-Antibiotic Resistance of Enterococcus faecalis and Enterococcus faecium from Cecal Contents in Broiler Chicken and Turkey Flocks Slaughtered in Canada and Plasmid Colocalization of tetO and ermB Genes

Journal of Food Protection ◽

10.4315/0362-028x.jfp-10-451 ◽

2011 ◽

Vol 74 (10) ◽

pp. 1639-1648 ◽

Cited By ~ 33

Author(s):

CINDY-LOVE TREMBLAY ◽

ANN LETELLIER ◽

SYLVAIN QUESSY ◽

MARTINE BOULIANNE ◽

DANIELLE DAIGNAULT ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Enterococcus Faecalis ◽

Resistance Genes ◽

Enterococcus Faecium ◽

Broiler Chicken ◽

Multidrug Resistant ◽

Poultry Meat ◽

Antimicrobial Resistance Genes ◽

High Level ◽

Level Resistance

This study was conducted to characterize the antimicrobial resistance determinants and investigate plasmid colocalization of tetracycline and macrolide genes in Enterococcus faecalis and Enterococcus faecium from broiler chicken and turkey flocks in Canada. A total of 387 E. faecalis and E. faecium isolates were recovered from poultry cecal contents from five processing plants. The percentages of resistant E. faecalis and E. faecium isolates, respectively, were 88.1 and 94% to bacitracin, 0 and 0.9% to chloramphenicol, 0.7 and 14.5% to ciprofloxacin, 72.6 and 80.3% to erythromycin, 3.7 and 41% to flavomycin, 9.6 and 4.3% (high-level resistance) to gentamicin, 25.2 and 17.1% (high-level resistance) to kanamycin, 100 and 94% to lincomycin, 0 and 0% to linezolid, 2.6 and 20.5% to nitrofurantoin, 3 and 27.4% to penicillin, 98.5 and 89.7% to quinupristin-dalfopristin, 7 and 12.8% to salinomycin, 46.7 and 38.5% (high-level resistance) to streptomycin, 95.6 and 89.7% to tetracycline, 73 and 75.2% to tylosin, and 0 and 0% to vancomycin. One predominant multidrug-resistant phenotypic pattern was identified in both E. faecalis and E. faecium (bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, tetracycline, and tylosin). These isolates were further examined by PCR and sequencing for the genes encoding their antimicrobial resistance. Various combinations of vatD, vatE, bcrR, bcrA, bcrB, bcrD, ermB, msrC, linB, tetM, and tetO genes were detected, and ermB, tetM, and bcrB were the most common antimicrobial resistance genes identified. For the first time, plasmid extraction and hybridization revealed colocalization of tetO and ermB genes on a ca. 11-kb plasmid in E. faecalis isolates, and filter mating experiments demonstrated its transferability. Results indicate that the intestinal enterococci of healthy poultry, which can contaminate poultry meat at slaughter, could be a reservoir for quinupristin-dalfopristin, bacitracin, tetracycline, and macrolide resistance genes.

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Presence and Characterization of a Novel cfr-Carrying Tn558 Transposon Derivative in Staphylococcus delphini Isolated From Retail Food

Frontiers in Microbiology ◽

10.3389/fmicb.2020.598990 ◽

2021 ◽

Vol 11 ◽

Author(s):

Feng Zhang ◽

Shi Wu ◽

Jiahui Huang ◽

Runshi Yang ◽

Jumei Zhang ◽

...

Keyword(s):

Public Health ◽

Multidrug Resistance ◽

Antimicrobial Resistance ◽

Resistance Gene ◽

Resistance Genes ◽

Poultry Meat ◽

Whole Genome ◽

Chloramphenicol Resistance ◽

Antimicrobial Resistance Genes ◽

Retail Food

Antimicrobial resistance has become a major public health threat. Food-related Staphylococcus species have received much attention due to their multidrug resistance. The cfr gene associated with multidrug resistance has been consistently detected in food-derived Staphylococcus species. In this retrospective study, we examined the prevalence of cfr-positive Staphylococcus strains isolated from poultry meat in different geographical areas of China from 2011 to 2016. Two cfr-positive Staphylococcus delphini strains were identified from poultry meat in China. Comparative and whole-genome analyses were performed to characterize the genetic features and overall antimicrobial resistance genes in the two S. delphini isolates 245-1 and 2794-1. Whole-genome sequencing showed that they both harbored a novel 20,258-bp cfr-carrying Tn558 transposon derivative on their chromosomes. The Tn558 derivative harbors multiple antimicrobial resistance genes, including the transferable multiresistance gene cfr, chloramphenicol resistance gene fexA, aminoglycoside resistance genes aacA-aphD and aadD, and bleomycin resistance gene ble. Surprisingly, within the Tn558 derivative, an active unconventional circularizable structure containing various resistance genes and a copy of a direct repeat sequence was identified by two-step PCR. Furthermore, core genome phylogenetic analysis revealed that the cfr-positive S. delphini strains were most closely related to S. delphini 14S03313-1 isolated from Japan in 2017 and 14S03319-1 isolated from Switzerland in 2017. This study is the first report of S. delphini harboring a novel cfr-carrying Tn558 derivative isolated from retail food. This finding raises further concerns regarding the potential threat to food safety and public health safety. The occurrence and dissemination of similar cfr-carrying transposons from diverse Staphylococcus species need further surveillance.

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Genotypic antimicrobial resistance characterization of E. coli from dairy calves at high risk of respiratory disease administered enrofloxacin or tulathromycin

Scientific Reports ◽

10.1038/s41598-020-76232-w ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

R. V. Pereira ◽

C. Foditsch ◽

J. D. Siler ◽

S. C. Dulièpre ◽

C. Altier ◽

...

Keyword(s):

High Risk ◽

Antimicrobial Resistance ◽

Respiratory Disease ◽

Resistance Genes ◽

Tetracycline Resistance ◽

Control Group ◽

E Coli ◽

Tetracycline Resistance Genes ◽

Antimicrobial Resistance Genes

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

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