Detection of genes encoding virulence factors and bacteriocins in fecal enterococci of poultry in Portugal

Seventy-six Enterococcus isolates (43 E. faecalis, 30 E. faecium, two E. durans, and one E. hirae) recovered from fecal samples of poultry in a slaughterhouse (one isolate per fecal sample and one fecal sample per lot of animals) were studied for bacteriocin production and for the presence of genes encoding bacteriocins and virulence factors. The presence of genes encoding virulence factors (cpd, geE, fsr, ace, agg, and esp) and bacteriocins (entA, entB, entP, entQ, entAS-48, entL50A/B, cyl, and bac31) were studied by polymerase chain reaction in all enterococci. At least two virulence genes were detected in all 43 E. faecalis isolates, cpd and gelE being the most frequently detected genes (97.7%) followed by ace (62.8%), agg (39.5%), fsr (27.9%), and esp (2.3%). No virulence genes were detected in the other enterococcal species with the exception of one E. faecium and one E. durans isolates that harbored the gelE gene. Antimicrobial activity against eight indicator bacteria (including Listeria monocytogenes) was assayed in the enterococci, and 23 (30.3%) showed inhibitory activity against L. monocytogenes, the other 22 enterococci showing activity against indicator bacteria other than L. monocytogenes. Only the entA, entB, and cyl genes were detected in our study (entA + entB in nine E. faecium isolates and the cyl gene in seven E. faecalis isolates). A wide variety of virulence genes have been detected in fecal E. faecalis isolates from poultry, but not in the other enterococcal species. However, the presence of known bacteriocin structural genes is associated more with the E. faecium species.     

Phenotypic and genotypic study of gelatinase and beta-haemolysis activities in faecal enterococci of poultry in Portugal

The detection of gelatinase and beta-haemolysis activities was carried out in 83 faecal enterococci (43 Enterococcus faecalis, 33 E. faecium, five E. durans and two E. hirae) of poultry origin. In addition, the presence of genes of the gelE-fsrABC locus and of the cyl operon (cylL(L), cylL(S), cylA, cylB and cylM) were studied by polymerase chain reaction and correlated with gelatinase and beta-haemolysis production, respectively. Most of our E. faecalis isolates were gelatinase-positive (88%), being this activity not frequent in the other enterococcal species (2.5%). Only one of the 33 E. faecium isolates showed a positive gelatinase reaction. All enterococci that showed gelatinase activity harboured the gelE and fsrABC genes, although these genes were also detected in four E. faecalis and one E. durans gelatinase-negative isolates. Most of our non-E. faecalis gelatinase-negative isolates did not harbour gelE-fsrABC genes. A high proportion of faecal enterococci of poultry origin harboured genes of the cyl operon (71%), although only 7% contained the five cyl tested genes (all of them E. faecalis). Only one isolate of our series could express beta-haemolysis, harbouring the whole cyl operon. The cylL(S) genotype was the most prevalent in our enterococci (39%) and also the most prevalent among our E. faecalis isolates (60%). Other genotypes detected were the following ones (% of enterococci): cylA + cylB + cylM (13%), cylL(L) + cylA (4%), cylL(L) (4%), cylL(L) + cylA + cylB + cylM (2%), cylL(L) + cylA + cylM (1%) and cylA + cylM (1%). Both phenotypic and genotypic assays are important to evaluate the virulence potential of enterococci.     

Occurrence of the structural enterocin A, P, B, L50B genes in enterococci of different origin

Enterococci are well-known producers of antimicrobial peptides--bacteriocins (enterocins) and the number of characterized enterocins has been significantly increased. Recently, enterocins are of great interest for their potential as biopreservatives in food or feed while research on enterocins as alternative antimicrobials in humans and animals is only at the beginning. The present study provides a survey about the occurrence of enterocin structural genes A, P, B, L50B in a target of 427 strains of Enterococcus faecium (368) and Enterococcus faecalis (59) species from different sources (animal isolates, food and feed) performed by PCR method. Based on our results, 234 strains possessed one or more enterocin structural gene(s). The genes of enterocin P and enterocin A were the most frequently detected structural genes among the PCR positive strains (170 and 155 strains, respectively). Different frequency of the enterocin genes occurrence was detected in strains according to their origin; the strains from horses and silage showed the highest frequency of enterocin genes presence. All possible combinations of the tested genes occurred at least twice except the combination of the gene of enterocin B and L50B which possessed neither strain. The gene of enterocin A was exclusively detected among E. faecium strains, while the gene of enterocin P, B, L50B were detected in strains of both species E. faecium and E. faecalis. In conclusion, a high-frequency and variability of enterocin structural genes exists among enterococci of different origin what offers a big possibility to find effective bacteriocin-producing strains for their application in veterinary medicine.     

Enterocin A结构基因和免疫基因的克隆及序列特征分析

采用PCR技术从屎肠球菌的染色体DNA上扩增到1条529 bp的DNA片段entAIc，以pGEM—T为载体，将该基因片段克隆到大肠杆菌中，对阳性重组质粒pGAI测序。序列分析结果表明：片段entAIc含有2个ORF，Enterocin A结构基因ent A和免疫基因entI，2个基因紧密相连。核苷酸序列分析显示，不同菌株来源的ent A和ent I具有较高的同源性，分别达到99．87％和99．76％。氨基酸序列分析显示不同菌株来源的Enterocin A前体完全相同，免疫蛋白仅有1个氨基酸差异。     

Acquired antimicrobial resistance in the intestinal microbiota of diverse cat populations

The aim of this study was to investigate the prevalence of acquired antimicrobial resistance in the resident intestinal microbiota of cats and to identify significant differences between various cat populations. Escherichia coli, Enterococcus faecalis, E. faecium and Streptococcus canis were isolated as faecal indicator bacteria from rectal swabs of 47 individually owned cats, 47 cattery cats and 18 hospitalised cats, and submitted through antimicrobial sensitivity tests. The results revealed that bacteria isolated from hospitalised and/or cattery cats were more frequently resistant than those from individually owned cats. E. coli isolates from hospitalised cats were particularly resistant to ampicillin, tetracycline and sulfonamide. Both enterococci and streptococci showed high resistance to tetracycline and in somewhat lesser extent to erythromycin and tylosin. Most E. faecium isolates were resistant to lincomycin and penicillin. One E. faecalis as well as one E. faecium isolate from hospitalised cats showed 'high-level resistance' (MIC > 500 microg/ml) against gentamicin, a commonly used antimicrobial agent in case of human enterococcal infections. The results of this research demonstrate that the extent of acquired antimicrobial resistance in the intestinal microbiota of cats depends on the social environment of the investigated population. It is obvious that the flora of healthy cats may act as a reservoir of resistance genes.     

Antimicrobial and heavy metal resistance in commensal enterococci isolated from pigs

Antibiotic resistance in animal isolates of enterococci is of public health concern because of the risk of transfer of antibiotic resistance isolates or resistance determinants to consumers via the food chain. In this study, phenotypic and genotypic resistance in 192 pig isolates of enterococci to ampicillin, avilamycin, avoparcin, bacitracin, flavophospholipol, gentamicin, narasin, tetracycline, tiamulin, tylosin, vancomycin, virginiamycin, copper and zinc were investigated by susceptibility test and molecular methods. Resistance rates varied between the species but all isolates were susceptible to ampicillin, avilamycin, avoparcin, gentamicin and narasin but resistant to tetracycline and tylosin and intermediately resistant to copper. Only Enterococcus gallinarum and Enterococcus casseliflavus were resistant to vancomycin and virginiamycin resistance was present in less than half the Enterococcus faecium isolates. Zinc resistance was largely confined to Enterococcus faecalis but bacitracin resistance was uncommon in E. faecalis in comparison with the other species. Tiamulin resistance was common in all species except E. casseliflavus. Resistance to flavophospholipol was detected in most E. faecium isolates and in a high proportion of E. gallinarum, E. casseliflavus and E. hirae/durans but was only found in one isolate of E. faecalis. No tetO, rplC, rplD, vanA, vanB, vatA and vatD genes were found. The presence of ermB, tetL, tetM, tcrB, aac6-aph2, tetK, tetS, vanC1, vanC2, lsaA, lsaB and vatE varied between the species and largely corresponded to the susceptibility phenotype. The findings show that resistance to antibiotics of high clinical significance for nosocomial Enterococcus infections is absent, whereas antimicrobial resistance was detected for some other antibiotics including bacitracin, flavophospholipol, tetracycline, tiamulin, tylosin and virginiamycin.     

Evaluation in vitro of the antagonistic substances produced by Lactobacillus spp. isolated from chickens

To determine the inhibitory capacity of lactic acid bacteria due to the action of antagonistic substances, we tested 474 isolates of Lactobacillus from the crop and cecum of chickens against gram-positive and gram-negative indicator microorganisms by the spot-on-the-lawn and well-diffusion antagonism methods. Of the 474 isolates, 265 demonstrated antimicrobial activity against the indicator microorganisms. Isolates identified as L. reuteri, L. salivarius, or Lactobacillus spp. inhibited Enterococcus faecalis, E. faecium, Listeria monocytogenes, and Salmonella spp. but not L. casei, L. delbrueckii, L. fermentum, or L. helveticus by the well-diffusion simultaneous antagonism method under anaerobic incubation conditions. The antagonistic substances produced by some of the Lactobacillus isolates were inactivated after treatment by proteolytic enzymes, which suggested that the substances could be antimicrobial peptides or bacteriocins.     

Characterization of antibiotic resistance genes and virulence factors in faecal enterococci of wild animals in Portugal

Antibiotic susceptibility was tested in 140 non-selected enterococci (73 Enterococcus faecalis, 45 E. faecium and 22 of other species) recovered from faecal samples of 77 wild animals in Portugal. Susceptibility testing for 11 antibiotics (vancomycin, teicoplanin, ampicillin, streptomycin, gentamicin, kanamycin, chloramphenicol, tetracycline, erythromycin, quinupristin-dalfopristin and ciprofloxacin) was determined by disk diffusion and agar dilution methods. Forty-four isolates (31.4%) showed susceptibility to all the antibiotics tested (5.5% of E. faecalis; 62.2% of E. faecium; and 78.6% of E. hirae). Neither ampicillin-resistance nor acquired-vancomycin-resistance was detected and 1.4% of the isolates showed high-level-resistance for gentamicin or streptomycin. Tetracycline and erythromycin resistances were shown in 28.6% and 20.1% of the isolates, respectively. Antibiotic resistance genes were studied by polymerase chain reaction (PCR) and sequencing and tet(M) + tet(L), erm(B) or aac(6')-aph(2') genes were detected in most of tetracycline-, erythromycin- or gentamicin-resistant enterococci respectively. Genes encoding virulence factors were studied by PCR and a wide variety of virulence genes were detected in most of E. faecalis isolates but were rarely found in E. faecium and not detected in the other species. The prevalence of genes encoding virulence factors in E. faecalis was as follows: cpd (98.6%), gelE (75.3%), agg (30.1%), fsr (17.8%), ace (9.6%) and esp (4.1%). Low percentages of antibiotic resistance was found in the faecal enterococci of wild animals but a wide variety of virulence genes were detected among E. faecalis isolates although were rare in the other species.     

我国18个省份猪源肠球菌耐药调查及分析

为研究我国猪源肠球菌的耐药情况,本研究采集18个省市猪养殖场的粪便样品836份进行了肠球菌的分离鉴定,并通过琼脂稀释法测定11种抗菌药物的最小抑菌浓度。结果显示:从836份样品中共分离出225株肠球菌,其中屎肠球菌106株,粪肠球菌56株,小肠肠球菌34株,鹑鸡肠球菌17株,铅黄肠球菌7株,坚忍肠球菌4株,Enterococcus thailandicus 1株。所有的肠球菌对被检抗菌药物耐药程度由高到低依次是四环素(98.2%)、阿米卡星(94.7%)、多西环素(89.3%)、米诺环素(88.9%)、氟苯尼考(88.4%)、红霉素(86.2%)、环丙沙星(63.1%)、氨苄西林(33.8%)、氯霉素(29.8%),未检测到万古霉素和利奈唑胺耐药菌株。除氨苄西林、万古霉素、利奈唑胺外,粪肠球菌对其余8种被检抗菌药物耐药率均高于屎肠球菌。小肠肠球菌对阿米卡星、红霉素、四环素、多西环素、米诺环素5种抗菌药物的耐药率高于屎肠球菌。肠球菌的多重耐药情况集中在6耐、7耐和8耐,占肠球菌总数的72.9%。本研究表明我国猪源肠球菌对常见抗菌药物耐药率较高,应引起更多关注。     

Prevalence of and resistance to anti-microbial drugs in selected microbial species isolated from bulk milk samples

The prevalence of strains of Staphylococcus aureus, coagulase-negative (CN) staphylococci, Listeria monocytogenes, Escherichia coli, Enterococcus faecalis, E. faecium and Bacillus cereus, was investigated in 111 bulk milk samples. Staphylococcus aureus was isolated from 38 samples, CN staphylococci from 63 samples, E. coli from 49 samples, E. faecalis or E. faecium from 107 samples, and L. monocytogenes from two samples. Bacillus cereus was not found in any of the samples and three samples were free of any of the selected species. Sensitivity to the anti-microbial drugs amikacin, ampicillin, ampicillin + sulbactam, cephalothin (CLT), cephotaxime, clindamycin, chloramphenicol (CMP), co-trimoxazole, erythromycin (ERY), gentamicin, neomycin, norfloxacin, oxacillin, penicillin, streptomycin (STR), tetracycline (TTC) and vancomycin was tested using the standard dilution technique. Minimum inhibitory concentration (MIC) characteristics (MIC50, MIC90, MIC range) were determined for each microbial species. Resistance against one or more anti-microbial drugs was found in 93% of S. aureus, 40% of CN staphylococci, 73% of E. coli, 88% of E.faecalis, 55% of E.faecium, and one L. monocytogenes strain. Most of the strains, particularly enterococci, were resistant to STR, TTC, and ERY (MIC50 4 microg/ml). A high percentage of staphylococci were resistant to beta-lactam antibiotics. High resistance to CLT was found in 11 strains of E. coli (MIC 256 microg/ml) and strains resistant to CMP (MIC90 16 microg/ml) were detected. The highest numbers of resistance phenotypes were found in E. coil (16) and CN staphylococci (12). Eighteen identical resistance phenotypes were demonstrated in indicator bacteria (E. coli, E. faecalis, E. faecium) and pathogens (S. aureus, CN staphylococci) isolated from the same bulk milk sample. The obtained resistance data were matched against the herd owners' information on therapeutic use of the drugs. This confrontation could not explain the findings of strains resistant to ERY or CMP. Our findings are evidence of selection of resistant strains among not only pathogenic agents, but also among indicator bacteria which can become significant carriers of transmissible resistance genes.     

Phenotypic and genotypic characterization of antimicrobial resistance in faecal enterococci from wild boars (Sus scrofa)

The objective was to study the prevalence of antimicrobial resistance and the mechanisms implicated in faecal enterococci of wild boars in Portugal. One hundred and thirty-four enterococci (67 E. faecium, 54 E. hirae, 2 E. faecalis, 2 E. durans and 9 Enterococcus spp.) were recovered from 67 wild boars (two isolates/sample), and were further analysed. High percentages of resistance were detected for erythromycin, tetracycline, and ciprofloxacin (48.5%, 44.8%, and 17.9%, respectively), and lower values were observed for high-level-kanamycin, -streptomycin, chloramphenicol, and ampicillin resistance (9%, 6.7%, 4.5%, and 3.7%, respectively). No isolates showed vancomycin or high-level-gentamicin resistance. The erm(B), tet(M), aph(3')-IIIa, and ant(6)-I genes were demonstrated in all erythromycin-, tetracycline-, kanamycin-, and streptomycin-resistant isolates, respectively. Specific genes of Tn916/Tn1545 and Tn5397 transposons were detected in 78% and 47% of our tet(M)-positive enterococci, respectively. The tet(S) and tet(K) genes were detected in one isolate of E. faecium and E. hirae, respectively. Three E. faecium isolates showed quinupristin-dalfopristin resistance and the vat(E) gene was found in all of them showing the erm(B)-vat(E) linkage. Four E. faecium isolates showed ampicillin-resistance and all of them presented seven amino acid substitutions in PBP5 protein (461Q-->K, 470H-->Q, 485M-->A, 496N-->K, 499A-->T, 525E-->D, and 629E-->V), in relation with the reference one; a serine insertion at 466' position was found in three of the isolates. Faecal enterococci from wild boars harbour a variety of antimicrobial resistance mechanisms and could be a reservoir of antimicrobial resistance genes and resistant bacteria that could eventually be transmitted to other animals or even to humans.     

产enterocin E5细菌素粪肠球菌发酵条件的优化

采用琼脂扩散法测定粪肠球菌发酵上清液对大肠埃希菌K88的抑菌活性,从而确定产enterocin E5细菌素粪肠球菌的发酵条件。结果表明,粪肠球菌E5产细菌素的发酵培养基为MRS培养基,最适温度为37℃,最适起始pH值6.5,最佳接种量2%,种龄14 h,发酵时间16 h,最佳培养基组分氮源为1%胰蛋白胨、0.5%酵母浸粉,最佳碳源为1%葡萄糖、0.5%蔗糖,0.1%Tween-80有利于enterocin E5的产生。这是分离自北京优良商品猪黑六产enterocin E5粪肠球菌的首次报道。     

2022年北京市宠物源细菌耐药性监测浅析

为了解北京市宠物源细菌的抗菌药物耐药情况,2022年,笔者对北京市四个城区的四家宠物医院的犬、猫共计50份样本的肛拭子进行了研究。试验对样品中的大肠杆菌和肠球菌首先进行了分离培养和质谱鉴定,然后采用微量肉汤稀释法分析分离菌株的耐药表型。结果共分离出大肠杆菌25株、肠球菌25株（屎肠球菌14株、粪肠球菌11株）。大肠杆菌耐药率最高的2种抗菌药为四环素和氨苄西林,多重耐药菌占44%;肠球菌耐药情况较严重,粪肠球菌耐药率最高的抗菌药物为磺胺异噁唑,屎肠球菌耐药率最高的抗菌药物为磺胺异噁唑、头孢西丁和红霉素,二者多重耐药菌占分离株总数的100%。综上,北京地区宠物源大肠杆菌、肠球菌的耐药情况较为严峻,且多重耐药现象突出,需要加强对宠物抗菌药使用的监督与管理。     

Genetic relatedness of high-level aminoglycoside-resistant enterococci isolated from poultry carcasses

Approximately 46% (75/162) or poultry enterococci collected between 1999 and 2000 exhibited high-level resistance to gentamicin (minimum inhibitory concentration [MIC] > or = 500 microg/ml), kanamycin (MIC > or = 500 microg/ml), or streptomycin (MIC > or = 1000 microg/ml). Forty-one percent of the isolates were resistant to kanamycin (n = 67), whereas 23% and 19% were resistant to genramicin (n = 37) and streptomycin (n = 31), respectively. The predominant species identified was Enterococcus faecium (n = 105), followed by Enterococcus faecalis (n = 40) and Enterococcus durans (n = 8). Using polymerase chain reaction, the isolates were examined for the presence of 10 aminoglycoside resistance genes [ant(6)-Ia, ant(9)-Ia, ant(4')-Ia, aph(3')-IIIa, aph(2")-Ib, aph(2")-Ic, aph(2")-Id, aac(6')-Ie-aph(2")-Ia, and aac(6')-Ii]. Five aminoglycoside resistance genes were detected, most frequently aac(6')-Ii and ant(6)-Ia from E. faecium. Seven E. faecalis isolates resistant to gentamicin, kanamycin, or streptomycin were negative for all genes tested, indicating that additional resistance genes may exist. Phylogenetic analysis revealed that the isolates were genetically different with little clonality. These data indicate that enterococci from poultry are diverse and contain potentially unidentified aminoglycoside resistance genes.     

猪源粪肠球菌和屎肠球菌多重PCR快速鉴定方法的建立

粪肠球菌和屎肠球菌是引起猪感染发病的优势肠球菌种,以肠球菌的16S rRNA基因设计属特异性引物,利用SodA基因多态性设计种特异性引物,同时优化反应条件,建立了能同时测定猪源粪肠球菌和屎肠球菌多重PCR方法。通过特异性和敏感性分析,该方法能扩增出肠球菌属特异性片段及粪肠球菌和屎肠球菌种特异性片段;对粪肠球菌敏感性为0...     

Effect of growth promotant usage on enterococci species on a poultry farm

The species population of enterococci isolated from four poultry houses for six grow-outs on one farm was determined. Two houses on the farm were control houses and did not use any antimicrobials, while two other houses on the farm used flavomycin, virginiamycin, and bacitracin during different poultry grow-outs. Litter, chick boxliners, feed, and poultry carcass rinses were obtained from each house and cultured for the presence of enterococci. Nine species of enterococci (Enterococcusfaecalis, E. faecium, E. avium, E. casselifiavus, E. cecorum, E. durans, E. gallinarum, E. hirae, and E. malodoratus) were identified from the study. Enterococcus faecalis was isolated more frequently from chick boxliners (n=176; 92%) and carcass rinses (n=491; 69%), whereas E. faecium was found more frequently in litter (n=361; 77%) and feed (n=67; 64%). Enterococcus faecalis (n=763; 52%) and E. faecium (n=578; 39%) were isolated most often from the farm and houses, regardless of antimicrobial treatment. Fifty-two percent of E. faecalis and 39% of E. faecium were isolated from both control (n=389 and 295, respectively) and treatment (n=374 and 283, respectively) samples. This study indicates that antimicrobial usage on this farm did not alter the resident population of enterococci.     

Isolation，Identification and Antibiotics Resistance Analysis of Enterococcus from Yak

17 feces samples of yak which were collected in Hongyuan county were measured with Gram staining method and 16S rRNA molecular identification in this study.8 suspected Enterococcus were separate from feces samples by bacteria purification and PCR amplification with 1 500 bp specific band. 6Enterococcus faecalis and 2Enterococcus faecium were identified through 16S rRNA sequencing.The homology analysis of the strains revealed that the homology between Enterococcus faecalis and reference strains sequence were 99.7% to 100%,that of Enterococcus faecium and reference sequence were 98.2% to 99.2%,indicating that the yak Enterococcus was highly conserved in the process of genetic evolution.The drug sensitive test results showed that the isolated strains were highly resistance to aminoglycoside antibiotics.Enterococcus faecium 11-1-2 strain was not only 5 multi-resistant,but also showed resistence to vancomycin.Enterococcus faecalis strains was most 3 multi-resistant.The antibiotics resistance results revealed that the resistance of yak Enterococcus was serious and should be taken seriously.     

牦牛源肠球菌的分离鉴定及耐药性分析

试验结合革兰氏染色及16S rRNA分子鉴定,对分离自红原县的17份牦牛粪便样中的肠球菌进行鉴定。结果显示,通过细菌分离纯化及PCR扩增,从牦牛粪便样品中分离出8株疑似肠球菌,分离菌的扩增产物经凝胶电泳后均产生1 500 bp特异性条带。16S rRNA测序结果显示,8株疑似肠球菌中,6株为粪肠球菌,2株为屎肠球菌。同源性比对分析显示,粪肠球菌与参考序列同源性为99.7%~100.0%,屎肠球菌与参考序列同源性为98.2%~99.2%,说明牦牛源肠球菌在遗传进化过程中高度保守。运用K-B纸片法进行药敏试验,结果显示,分离株对氨基糖苷类抗生素耐受性较高,2株屎肠球菌中,11-1-2菌株表现为5重耐药,且该菌株耐万古霉素,粪肠球菌主要表现为3重耐药,药敏结果提示牦牛源肠球菌耐药较严重,应引起高度重视。     

Monitoring of antimicrobial resistance in healthy dogs: first report of canine ampicillin-resistant Enterococcus faecium clonal complex 17

National surveillance programs on antimicrobial usage and antimicrobial resistance in animals have been established in various countries but few of them include bacteria from pets. The objectives of this study were to assess the prevalence of antimicrobial resistance in healthy dogs and to search for resistance phenotypes of clinical relevance. Escherichia coli and Enterococcus spp. were isolated from faecal swabs obtained from 127 dogs. Disk diffusion was used to measure antimicrobial susceptibility in 117 Escherichia coli, 10 Enterococcus faecium and 51 Enterococcus faecalis of canine origin. Resistance was relatively low compared with food animal species in Denmark. All Escherichia coli isolates were susceptible to broad-spectrum aminopenicillins, third generation cephalosporins and fluoroquinolones. Despite the low prevalence of resistance, statistical analysis of questionnaire data revealed a significant association (p=0.02) between recent antimicrobial treatment and resistance in Escherichia coli. Interestingly, two dogs were found to shed Enterococcus faecium resistant to ampicillin. Multilocus sequence typing of these isolates indicated that the two isolates belonged to sequence types associated with human nosocomial infections, and one (ST-192) was genetically related to human epidemic clonal complex 17. The detection of ampicillin-resistant Enterococcus faecium warrants further studies on the prevalence of these bacteria in dogs and on the possible implications to both animal and human health. The results suggest that distinct methods for detection and assessment of antimicrobial resistance in animals should be considered depending on the target animal species and the purposes of the study.     

Antimicrobial susceptibility and distribution of antimicrobial-resistance genes among Enterococcus and coagulase-negative Staphylococcus isolates recovered from poultry litter

Data on the prevalence of antimicrobial resistant enterococci and staphylococci from the poultry production environment are sparse in the United States. This information is needed for science-based risk assessments of antimicrobial use in animal husbandry and potential public-health consequences. In this study, we assessed the susceptibility of staphylococci and enterococci isolated from poultry litter, recovered from 24 farms across Georgia, to several antimicrobials of veterinary and human health importance. Among the 90 Enterococcus isolates recovered, E. hirae (46%) was the most frequently encountered species, followed by E. faecium (27%), E. gallinarum (12%), and E. faecalis (10%). Antimicrobial resistance was most often observed to tetracycline (96%), followed by clindamycin (90%), quinupristin-dalfopristin (62%), penicillin (53%), erythromycin (50%), nitrofurantoin (49%), and clarithromycin (48%). Among the 110 staphylococci isolates recovered, only coagulase-negative staphylococci (CNS) were identified with the predominant Staphylococcus species being S. sciuri (38%), S. lentus (21%), S. xylosus (14%) and S. simulans (12%). Resistance was less-frequently observed among the Staphylococcus isolates for the majority of antimicrobials tested, as compared with Enterococcus isolates, and was primarily limited to clarithromycin (71%), erythromycin (71%), clindamycin (48%), and tetracycline (38%). Multidrug resistance (MDR) phenotypes were prevalent in both Enterococcus and Staphylococcus; however, Enterococcus exhibited a statistically significant difference in the median number of antimicrobials to which resistance was observed (median = 5.0) compared with Staphylococcus species (median = 3.0). Because resistance to several of these antimicrobials in gram-positive bacteria may be attributed to the shuttling of common drug-resistance genes, we also determined which common antimicrobial-resistance genes were present in both enterococci and staphylococci. The antimicrobial resistance genes vat(D) and erm(B) were present in enterococci, vgaB in staphylococci, and mobile genetic elements Tn916 and pheromone-inducible plasmids were only identified in enterococci. These data suggest that the disparity in antimicrobial-resistance phenotypes and genotypes between enterococci and staphylococci isolated from the same environment is, in part, because of barriers preventing exchange of mobile DNA elements.