Comparative Analysis of Chloramphenicol-Resistant Enterococcus faecalis Isolated from Dairy Companies in Korea.

Although chloramphenicol is currently banned from use in livestock, other phenicols, such as florfenicol and thiamphenicol, have been used for the treatment of bacterial infections in domestic cattle in Korea. This study compares the characteristics of chloramphenicol-resistant Enterococcus faecalis isolated from the bulk tank milk of four major dairy companies in Korea. Although the distribution of multidrug resistance patterns showed no significant differences between the four companies, 85 chloramphenicol-resistant Enterococcus faecalis isolates showed a significantly high number of resistances against five or six antimicrobial classes (37.6%, respectively) (p < 0.05). When analyzing the distribution of phenicol resistance genes, 31 (36.5%) isolates only carried the catA gene, and two (6.3%) isolates from company A only carried the cfr gene. No isolates carried the catB or fexA genes. Regarding the distribution of other resistance genes, both the tetL and tetM (45.9%), ermB (82.4%), and both aac(6″)-Ie-aph(2″)-la and ant(6')-Ia genes (30.6%) showed a high prevalence, and the optrA and poxtA genes were observed separately, each in only two (2.4%) isolates. Our results confirm that the dissemination of chloramphenicol-resistant Enterococcus faecalis and some antimicrobial resistance genes show significant differences between dairy companies. Therefore, our results support that each dairy company should undertake effective surveillance programs to better understand and minimize the emergence of resistance on a multidisciplinary level.

Detection of Linezolid-Resistant Enterococcus faecalis and Enterococcus faecium Isolates from the Layer Operation System in Korea.

Linezolid (LNZ) is one of the most important antimicrobial agents against infections caused by gram-positive bacteria, including enterococci. In a layer operation system, antimicrobial resistance can be transferred to commercial layers via the fecal-oral route. This study investigated the presence and distribution of LNZ-resistant Enterococcus faecalis and Enterococcus faecium in a layer operation system. Among 117 E. faecalis and 154 E. faecium, 10 (8.5%) E. faecalis and 5 (3.2%) E. faecium isolates showed resistance to LNZ and chloramphenicol, and they exhibited multidrug resistance against 5 or more classes of antimicrobial agents. Among the resistant isolates, 9 (90.0%) and 2 (20.0%) E. faecalis harbored optrA and cfr genes, respectively. The optrA and fexA genes were not detected in five LNZ-resistant E. faecium. None of the 15 LNZ-resistant isolates harbored the fexA gene, and no mutations were observed in the genes encoding domain V of 23S ribosomal RNA (rRNA) and ribosomal proteins L3 (rplC) and L4 (rplD). Transferability was identified in three of the nine optrA-positive LNZ-resistant isolates. The tetM, tetL, and ermB genes were cotransferred with the optrA gene in all optrA-positive transconjugants. The results indicate that optrA is well-distributed in E. faecalis, implying a greater level of transferability. Thus, enhanced surveillance efforts are needed to monitor the emergence and spread of optrA in enterococci in layer operation system.

Virulence Variation of Salmonella Gallinarum Isolates through SpvB by CRISPR Sequence Subtyping, 2014 to 2018.

Salmonella Gallinarum is a Gram-negative bacteria that causes fowl typhoid, a septicemic disease with high morbidity and mortality that affects all ages of chickens. Although vaccines and antimicrobials have been used nationwide to eradicate the disease, the malady is still prevalent in Korea. In this study, we investigated the virulence and genetic variation of 116 S. Gallinarum isolates from laying hens between 2014 and 2018. A total of 116 isolates were divided into five Gallinarum Sequence Types (GST) through clustered regularly interspaced short palindromic repeats (CRISPR) subtyping method. The GSTs displayed changes over time. The 116 isolates showed no difference in virulence gene distribution, but the polyproline linker (PPL) length of the SpvB, one of the virulence factors of Salmonella spp., served as an indicator of S. Gallinarum pathogenicity. The most prevalent PPL length was 22 prolines (37.9%). The shortest PPL length (19 prolines) was found only in isolates from 2014 and 2015. However, the longest PPL length of 24 prolines appeared in 2018. This study indicates that PPLs of S. Gallinarum in Korea tend to lengthen over time, so the pathogenic potency of the bacteria is increasing. Moreover, the transition of GST was associated with PPL length extension over time. These results indicate that surveillance of changing GST and PPL length are necessary in the monitoring of S. Gallinarum isolates.

Characteristics of linezolid-resistant Enterococcus faecalis isolates from broiler breeder farms.

Linezolid is an oxazolidinone class antibiotic used for treatment infections caused by various multidrug-resistant gram-positive pathogens including enterococci. However, recently, linezolid-resistant isolates in animals are considered as a human health hazard. In a broiler operation system, antimicrobial resistance can be transferred to the environment and commercial broiler via the fecal-oral route. Therefore, this study was conducted to investigate the prevalence and characteristics of linezolid-resistant Enterococcus faecalis (E. faecalis) from broiler parent stock in a broiler operation system. Among 297 E. faecalis isolates from 85 flocks in 8 broiler breeder farms, the prevalence of chloramphenicol- and linezolid-resistant isolates was 0 to 12.1% and 0 to 8.0%, respectively; however, there were no significant differences between farms. Therefore, a total of 14 (4.7%) chloramphenicol- and/or linezolid-resistant E. faecalis showed resistance to 7 or more antimicrobial classes. The drug-resistance gene optrA, which can confer resistance to linezolid, tedizolid, and phenicols, was found in 8 (2.69%) isolates, and 7 (2.36%) of the 8 optrA-positive isolates co-carried the phenicol exporter gene fexA. However, E. faecalis isolates from 3 of 8 broiler breeder farms only carried the optrA and/or fexA genes. As linezolid is one of the last antimicrobial treatments of choice for multidrug-resistant gram-positive pathogens including E. faecalis, the presence of antibiotic-resistant E. faecalis in broiler breeder farms should be monitored to prevent the introduction of linezolid-resistant strains to the food chain.

Molecular characteristics of optrA-carrying Enterococcus faecalis from chicken meat in South Korea.

The purpose of this study was to identify the genetic environment of optrA gene in linezolid (LZD)-resistant Enterococcus faecalis from chicken meat and to describe the probable mechanism of dissemination of the optrA gene through plasmid or chromosomal integration. Whole genome sequencing and analysis revealed that all 3 E. faecalis isolates confirmed as LZD- and chloramphenicol-resistant carried fexA adjacent to the optrA gene as well as a variety of resistance genes for macrolides, tetracyclines, and aminoglycosides, simultaneously. But, the other genes conferring LZD resistance, cfr and poxtA, were not detected in those strains. Two isolates harboring the optrA gene in their chromosomal DNA showed >99% similarity in arrangement to the transposon Tn6674 and the transposase genes, tnpA, tnpB, and tnpC and were located in the first open reading frame for transposase. One isolate harboring an optrA-carrying plasmid also showed >99% similarity with the previously reported pE439 plasmid but had 2 amino acid changes (Thr96Lys and Tyr160Asp) and a higher minimum inhibitory concentration against LZD of 16 mg/L than that of pE439 (8 mg/L). Mobile genetic elements such as transposons or plasmids flanking the optrA gene conduct a crucial role in the dissemination of antimicrobial resistance genes. Further investigations are required to identify the way by which optrA is integrated into chromosomal DNA and plasmids.

Molecular characteristics of fluoroquinolone-resistant avian pathogenic Escherichia coli isolated from broiler chickens.

Avian pathogenic Escherichia coli (APEC) is a major pathogen in the poultry industry worldwide including Korea. In this study, the phenotypic and genotypic characteristics of 33 fluoroquinolone (FQ)-resistant APEC isolates from broilers were analyzed. All FQ-resistant APEC isolates showed amino acid exchanges at both gyrA and parC and high minimal inhibitory concentrations for FQs. A total of 11 (33.3%) isolates were positive for the plasmid-mediated quinolone resistance (PMQR) genes, qnrA (8 isolates) and qnrS (3 isolates), and showed multidrug resistance. Among the 11 PMQR-positive isolates, 1 and 2 isolates carried blaCTX-1 and blaCTX-15, respectively, as extended-spectrum ß-lactamase (ESBL) producers, and the non-ESBL gene, blaTEM-1, was found in 4 isolates. Among 3 aminoglycoside-resistant isolates, aac(3)-II was only detected in 1 isolate. All 8 APEC isolates with resistance to tetracycline carried the tetA gene. Overall, 6 of the 7 trimethoprim-sulfamethoxazole-resistant isolates carried the sul1 or sul2 genes, while only 2 of the 8 chloramphenicol-resistant isolates carried the catA1 gene. Although 9 isolates carried class I integrons, only 4 isolates carried the gene cassettes dfrA12-aadA2 (2 isolates), dfrA17-aadA5 (1 isolate), extX-psp-aadA2 (1 isolate), and dfrA27 (1 isolate). The most common plasmid replicon was FIB (8 isolates, 72.7%), followed by K/B (4 isolates, 36.4%). Antimicrobial resistance monitoring and molecular analysis of APEC should be performed continuously to surveil the transmission between poultry farms.

Antimicrobial resistance monitoring of commensal Enterococcus faecalis in broiler breeders.

Enterococcus faecalis (E. faecalis) has rapidly acquired resistance to multiple antimicrobials, and the antimicrobial resistance of E. faecalis from broiler breeders has been implicated in its vertical transmission to their offspring. The objective of this study was to investigate the antimicrobial resistance and genetic diversity of commensal E. faecalis isolated from the broiler breeder farms. Among a total of 229 E. faecalis isolates from 9 broiler breeder farms, the highest resistance rate was observed in tetracycline (78.2%), followed by doxycycline (58.1%) and erythromycin (43.7%), and the prevalence of antimicrobial resistance showed significant differences among the 9 broiler breeder farms (P < 0.05). The tetM gene (77.1%) and ermB gene (85.0%) were detected at the highest levels in 179 TE-and 100 E-resistant isolates, respectively. Twenty-four high-level gentamicin-resistant isolates carried aac(6″)Ie-aph(2″)-la gene, and 9 high-level ciprofloxacin-resistant isolates showed point mutations in both gyrA and parC genes. All high-level gentamicin-resistant or high-level ciprofloxacin-resistant isolates showed one of the two different virulence gene patterns, ace-asa1-efaA-gelE complex or ace-efaA-gelE complex. These results indicate that constant epidemiological monitoring at the breeder level is required to prevent the pyramidal transmission of antimicrobial-resistant E. faecalis.

Impacts and characteristics of antimicrobial resistance of Escherichia coli isolates by administration of third-generation cephalosporins in layer hatcheries.

We investigated the characteristics and persistence of Escherichia coli resistant to third-generation cephalosporins (3GCs) by early administration of ceftiofur or gentamicin and to analyze the impact of 3GC use in hatcheries. We studied 10 ceftiofur-treated flocks (CTFs) and 10 gentamicin-treated flocks (GTFs) of layers. Fecal samples were collected at 1, 2, 4, 8, 18, and 30 weeks of age for all flocks. Among the 446 E. coli isolates, 58 (29.0 %) of 200 isolates in CTFs were identified as 3GC-resistant E. coli and 28 (11.4 %) of 246 isolates in GTFs were identified as 3GC-resistant E. coli. The presence of 3GC-resistant E. coli isolates at 1, 2, and 4 weeks was significantly higher in CTFs than in GTFs (p < 0.05). Moreover, the rate of resistance to 3GCs gradually decreased from 83.3 % at 1 week of age to 4.4 % at 30 weeks of age in CTFs. Of the 86 3GC-resistant E. coli isolates, 32 isolates had ß-lactamase-encoding gene: blaCTX-M-14 (ten isolates), blaCTX-M-15 (three isolates), blaCMY-2 (five isolates), and blaTEM-1 (twenty-five isolates) genes. Plasmid replicon typing revealed that blaCTX-M-14, blaCTX-M-15, blaCMY-2, and blaTEM-1 were located on F, F and FIB, I1 and K, and I1 and FII, respectively. Furthermore, 18 isolates carried class 1 integrons, with four different gene cassettes. These results revealed that ceftiofur used in hatcheries can lead to an increase in the number of 3GC-resistant E. coli with many characteristics. A voluntary ban must be imposed on the use of 3GCs for 1-day-old chicks in poultry industry.

Molecular characterization of avian pathogenic Escherichia coli from broiler chickens with colibacillosis.

Avian pathogenic Escherichia coli (APEC) causes extensive mortality in poultry flocks, leading to extensive economic losses. The aim of this study was to investigate the phenotypic and genotypic characteristics and antimicrobial resistance of recent APEC isolates. Of the 79 APEC isolates, the most predominant serogroup was O78 (16 isolates, 20.3%), followed by O2 (7 isolates, 8.9%) and O53 (7 isolates, 8.9%). Thirty-seven (46.8%) and six (7.6%) of the isolates belonged to phylogenetic groups D and B2, respectively, and presented as virulent extraintestinal E. coli. Among 5 analyzed virulence genes, the highest frequency was observed in hlyF (74 isolates, 93.7%), followed by iutA (72 isolates, 91.9%) gene. The distribution of the iss gene was significantly different between groups A/B1 and B2/D (P < 0.05). All group B2 isolates carried all 5 virulence genes. APEC isolates showed high resistance to ampicillin (83.5%), nalidixic acid (65.8%), tetracycline (64.6%), cephalothin (46.8%), and ciprofloxacin (46.8%). The ß-lactamases-encoding genes blaTEM-1 (23 isolates, 29.1%), blaCTX-M-1 (4 isolates, 5.1%), and blaCTX-M-15 (3 isolates, 3.8%); the aminoglycoside-modifying enzyme gene aac(3)-II (4 isolates, 5.1%); and the plasmid-mediated quinolone genes qnrA (10 isolates, 12.7%) and qnrS (2 isolates, 2.5%) were identified in APEC isolates. The tetA (37 isolates, 46.8%) and sul2 (20 isolates, 25.3%) were the most prevalent among tetracycline and sulfonamide resistant isolates, respectively. This study indicates that APEC isolates harbor a variety of virulence and resistance genes; such genes are often associated with plasmids that facilitate their transmission between bacteria and should be continuously monitored to track APEC transmission in poultry farms.

Antimicrobial Resistance and Genetic Diversity of Salmonella Serotypes Recovered from Edible Pork Offal from Korea.

Although edible offal can be easily contaminated with foodborne bacteria and regarded as important transfers of antimicrobial resistance to humans, the characterization of bacteria from edible offal have not been researched sufficiently. This study is the first to focus on the molecular characterization of Salmonella isolated from edible pork offal. From a total of 52 Salmonella isolates from edible pork offal, 44 (80.7%) were resistant to at least one antimicrobial agent and 24 (46.2%) exhibited multidrug resistance (MDR). All MDR Salmonella were also resistant to ß-lactams and 12 (50.0%) of the isolates were positive for blaTEM-1. Eleven (68.8%) of the 16 gentamicin-resistant isolates harbored the ant(2'')-I gene. Among 18 tetracycline-resistant isolates, tetA and tetB genes were found in 9 (50.0%) and 3 (16.7%) isolates, respectively. The sul1 gene was identified in 9 (81.8%) of 11 trimethoprim/sulfamethoxazole-resistant isolates, and the cmlA gene was identified in only 2 (18.1%) among 11 chloramphenicol-resistant isolates. Eighteen (75.0%) of the 24 MDR Salmonella were identified as containing class 1 integrons, within which dfrA12-aadA2 (55.6%) was the most prevalent resistance gene cassettes. Twenty-one (87.5%) of the MDR isolates were also found to have the plasmid replicons. Replicon B/O (41.7%) was the most prevalent replicon types. These results suggest that edible pork offal can become a reservoir that not only harbors MDR Salmonella, but also contributes to their dissemination through cross-contamination processes.

Genetic characterization of high-level aminoglycoside-resistant Enterococcus faecalis and Enterococcus faecium isolated from retail chicken meat.

Retail chicken meat can play a role in the transfer of drug resistance to humans through the handling or ingestion of improperly cooked meat contaminated with resistant enterococci. In fact, high-level aminoglycoside-resistance (HLAR) in enterococci identified in human cases. Therefore, the prevalence and genetic characterization of HLAR in enterococci in retail chicken meat were investigated in this study. Of the 345 enterococci strains, 29 (8.7%) showed HLAR. All HLAR in enterococci carried at least 1 of 2 aminoglycoside-modifying enzyme genes, aac(6')Ie-aph(2″)-Ia and ant(6)-Ia. Among the 13 isolates that carried aac(6')Ie-aph(2″)-Ia, 3 had pattern A, with IS256 at both ends, and the other 10 had pattern D, without IS256 at both ends. All HLAR in enterococci also showed multidrug resistance. Among the 24 erythromycin-resistant enterococci, 19 (79.2%) harbored the ermB gene, and one (4.2%) harbored both the ermB and ermA genes. A total of 21 enterococci were tetracycline-resistant and harbored one or more of the following tetracycline resistance genes tet(M), tet(L), and tet(O). The Int-Tn gene was detected in one isolate (3.4%) carrying the tet(M) and ermB genes. All 4 chloramphenicol-resistant isolates carried either the phenicol resistance gene cfr alone (one isolate), both cfr and fexA (one isolate), or both fexA and optrA (2 isolates). Four efflux pump genes, efr(A), efr(B), emeA, and lsa, were detected in all HLAR in Enterococcus faecalis isolates. These results improve our understanding of the transmission dynamics of HLAR in enterococci from non-hospital sources to humans.

Molecular Characterization of Multidrug-Resistant Escherichia coli Isolates from Edible Offal in Korea.

HIGHLIGHTS: Edible offal is significantly contaminated by antimicrobial-resistant Escherichia coli. E. coli from edible offal is harboring various antimicrobial resistance and virulence genes. Improvements in hygienic conditions of edible offal production is required.

Molecular characterization of antimicrobial-resistant Enterococcus faecalis and Enterococcus faecium isolated from layer parent stock.

Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) are ubiquitous intestinal bacteria in humans and animals that can easily acquire antimicrobial resistance, which allows them to have roles as antimicrobial resistance indicators. In addition, layer parent stock produces thousands of eggs for the production of commercial laying hens and can transfer a variety of viral and bacterial agents to chicks. The objective of this study was to determine the prevalence and characteristics of antimicrobial-resistant E. faecalis and E. faecium isolated in the layer parent stock level of the egg-layer operating system in South Korea. A total of 129 E. faecalis and 166 E. faecium isolates from 74 flocks of 30 layer parent stock were tested for resistance in this study. The prevalence of doxycycline- (51.9%), erythromycin- (53.5%), high-level gentamicin- (13.2%), high-level kanamycin- (31.0%), high-level streptomycin- (30.2%), and tetracycline- (64.3%) resistant E. faecalis isolates were higher than those for E. faecium isolates (P < 0.05). The ermB gene was detected in 66 (95.7%) erythromycin-resistant E. faecalis isolates, which was higher than that of 32 (71.7%) erythromycin-resistant E. faecium isolates. Twenty-one high-level gentamicin-resistant Enterococcus spp. (17 E. faecalis and 4 E. faecium) carried at least one aminoglycoside-modifying enzyme gene, aac(6')Ie-aph(2')-Ia or ant(6)-Ia. Fourteen isolates that harbored both aac(6')Ie-aph(2')-Ia and ant(6)-Ia exhibited pattern A with IS256 at both ends. Ten high-level ciprofloxacin-resistant Enterococcus spp. (8 E. faecalis and 2 E. faecium) showed amino acid changes from serine to isoleucine at codons 83 in gyrA, and 80 in parC. Also, the virulence genes ace, asa1, efaA, and gelE were detected in this study. To the best of our knowledge, this is the first study to examine the prevalence and characteristics of antimicrobial-resistant E. faecalis and E. faecium isolates in the layer parent stock. Our findings support the need for a surveillance program to monitor the emergence of antimicrobial-resistant E. faecalis and E. faecium in layer operating system.

Molecular characteristics of extended-spectrum and plasmid-mediated AmpC ß-lactamase-producing Escherichia coli isolated from commercial layer in Korea.

In the poultry industry, commercial layer farms play an important role in meeting the protein demand through the supply of eggs. However, the risk of contamination by ß-lactamase-producing Escherichia coli in eggs laid by commercial chickens is significant. In this study, we investigated the rate of extended-spectrum ß-lactamase (ESBL) and plasmid-mediated AmpC (pAmpC) ß-lactamase-producing E. coli isolated from layer hens and characterized their molecular background. Among the 92 cefotaxime-resistant E. coli isolates, 66 (71.7%) were identified as multidrug resistant and 29 showed phenotypic and genotypic characteristics of ß-lactamase-producing E. coli. The ESBL/pAmpC genes blaCTX-M-1, blaCTX-M-14, blaCTX-M-15, and blaCMY-2 were detected in 1, 6, 5, and 4 isolates, respectively. The non-ESBL/pAmpC gene blaTEM-1 was found in 16 isolates. Three isolates harbored both blaTEM-1 and blaCTX-M-14 genes. A total of 12 isolates also carried class 1 integrons, with 3 different gene cassette arrangements found in 8 of these isolates. A pulsed-field gel electrophoresis (PFGE) analysis of the 29 ß-lactamase-producing E. coli isolates revealed that 4 PFGE patterns were consistent with the ß-lactamase gene and layer farm origin, and showed a similar antibiotic resistance pattern. Our results suggest that comprehensive surveillance and more prudent use of third-generation cephalosporins in commercial layer farms is necessary to prevent the dissemination of ESBL/pAmpC-producing E. coli.

Characteristics of cephalosporin-resistant Salmonella isolates from poultry in Korea, 2010-2017.

Cephalosporins are the key drugs for the treatment of salmonellosis. Resistance to cephalosporins in Salmonella spp. has become a serious public health concern worldwide. Although the sales of cephalosporins have increased by five times from 2008 to 2016 in Korea, limited information is available on cephalosporin resistance in Korea. Therefore, we aimed to investigate the trends in prevalence and characteristics of cephalosporin-resistant Salmonella isolates from poultry in Korea in the period between 2010 and 2017. A total of 141 Salmonella isolates were collected from various poultry industry sources, and 38 (27.0%) among them showed resistance to cephalosporins. In particular, resistance to the following cephalosporins increased significantly over the seven-year period cephalothin (from 5.0% to 29.2%), cephalexin, cefuroxime, cefotaxime, and ceftazidime (from 0% to 25.0%), and cefepime (from 0% to 12.5%). In addition, 12 isolates carried a ß-lactamase gene. A non-extended-spectrum ß-lactamase (ESBL) and plasmid-mediated AmpC ß-lactamase (pAmpC) gene, blaTEM-1, was found in three isolates in the periods of 2010 to 2011 and 2012 to 2013, respectively. The blaCTX-M-79 (n = 4) and blaCTX-M-15 (n = 1) for ESBL genes and blaCMY-2 (n = 1) for pAmpC genes were only present in the 2016-2017 period. All ESBL/pAmpC-positive isolates had high minimum inhibitory concentrations for most cephalosporins and showed multi-drug resistance. In a conjugation experiment, the transfer of blaCTX-M-79 and blaCMY-2 genes was confirmed in transconjugants, which showed similar pattern of antibiotic resistance. This demonstrates that ESBL/pAmpC-producing Salmonella isolates might be transmitted to humans through contaminated poultry products. These findings suggest the need for the development of monitoring program and the guidelines for prudent use of antimicrobial agents in the poultry industry in Korea.

Characteristics of third-generation cephalosporin-resistant Salmonella from retail chicken meat produced by integrated broiler operations.

Integrated broiler operations, which control and operate vertically through all phases of the chicken industry, have applied biosecurity and sanitation practices, housing technologies, feeding regimens, and antibiotic applications in different ways to improve food safety. The objective of this study was to compare the prevalence and antimicrobial resistance profiles of Salmonella isolates recovered from 6 different integrated broiler operations and to analyze the characteristics of extended-spectrum ß-lactamase (ESBL)- and plasmid-mediated AmpC ß-lactamase (pAmpC)-producing Salmonella isolates. Among 336 chicken meat samples, 57 were observed to be positive for Salmonella. However, the prevalence varied from 6.8% to 45.8% in chicken meat, indicating variations in Salmonella occurrence among the operations. Salmonella Albany was the dominant serovar, followed by Salmonella Virchow. In the antimicrobial resistance test, nalidixic acid-resistant isolates were the most prevalent (73.7%), followed by isolates resistant to ampicillin (49.1%) and tetracycline (42.1%). Among 14 third-generation cephalosporin-resistant isolates, 9 (64.3%) ESBL/pAmpC-producing isolates were only obtained from 2 operations: blaCTX-M-15 (n = 7) and blaCTX-M-79 (n = 1) for ESBL genes and blaCMY-2 (n = 1) for pAmpC. All ESBL/pAmpC-positive isolates exhibited high minimum inhibitory concentrations (≥128 µg/mL) of most cephalosporins and showed multidrug resistance. The transfer of ESBL/pAmpC genes was confirmed in transconjugants, which had the same genes and similar resistance patterns as those of the donor. Our findings suggest that Salmonella with resistance to third-generation cephalosporins can now be found in association with integrated broiler operations, providing data to support the development of monitoring and prevention programs for the development and spread of antimicrobial resistance in integrated broiler operations.

Molecular characterization of erythromycin and tetracycline-resistant Enterococcus faecalis isolated from retail chicken meats.

Enterococcus faecalis is a ubiquitous intestinal bacterium in human and animals that can easily acquire antimicrobial resistance, which allows it to play the role of an antimicrobial resistance indicator. The objectives of this study were to characterize erythromycin and tetracycline-resistant E. faecalis isolated from retail chicken meats. A total of 149 among 335 E. faecalis isolates from 7 integrated broiler operations showed the simultaneous resistance to erythromycin and tetracycline, and more than 50% among 149 isolates showed multidrug resistance. The most common resistance genes were ermB (96.0%, 143 isolates) related with macrolides resistance, and tet(M) (95.3%, 142 isolates) and tet(L) (89.3%, 133 isolates) related with tetracycline resistance. Furthermore, 140 (93.9%) isolates simultaneously possessed ermB, and tet(L) and/or tet(M) genes. Eight isolates with transposon of the Tn916/1545-like were detected, which also carried ermB and tet(M) genes. The most prevalent of virulence genes were gelE (142 isolates, 95.3%), ace (137 isolates, 91.9%), and efaA (120 isolates, 80.5%). Also, 5 E. faecalis isolates successfully transferred antimicrobial and virulence genes to E. faecalis FA2-2. Therefore, the antimicrobial-resistant E. faecalis isolates as well as their corresponding genes and mobile genetic elements, such as transposons may be disseminated nationwide by broiler operation system in Korea.

Characteristics of High-Level Ciprofloxacin-Resistant Enterococcus faecalis and Enterococcus faecium from Retail Chicken Meat in Korea.

Genes encoding ciprofloxacin resistance in enterococci in animals may be transferred to bacteria in the animal gut and to zoonotic bacteria where they could pose a human health hazard. The objective of this study was to characterize antimicrobial resistance in high-level ciprofloxacin-resistant (HLCR) Enterococcus faecalis and Enterococcus faecium isolated from retail chicken meat. A total of 345 enterococci (335 E. faecalis and 10 E. faecium) were isolated from 200 chicken meat samples. Of these, 85 E. faecalis isolates and 1 E. faecium isolate were confirmed as HLCR enterococci. All 86 HLCR enterococci displayed gyrA- parC point mutations consisting of S83I-S80I (94.2%, 81 isolates), S83F-S80I (2.3%, 2 isolates), S83Y-S80I (2.3%, 2 isolates), and S83Y-S80F (1.2%, 1 isolate). Sixty-one (72.9%) of the 86 HLCR enterococci showed multidrug resistance to three to six classes of antimicrobial agents. Multilocus sequence typing revealed that E. faecalis had 17 different sequence types (ST) and E. faecium had 1 different ST, with ST256 observed most often (44 isolates, 51.8%). Although these results cannot exclude the possibility that pathotypes of enterococci isolated from chicken might represent transmission to or from humans, the foodborne HLCR E. faecalis indicated that the food chain is a potential route of enterococcal infection in humans.

Comparative genetic characterization of third-generation cephalosporin-resistant Escherichia coli from chicken meat produced by integrated broiler operations in South Korea.

Vertical integration of the broiler industry allows producers to combine different biosecurity and sanitation practices, housing technologies, and feeding regimens to improve food safety. The objectives of this study were to determine the antimicrobial resistance pattern of ß-lactamase-producing E. coli and to compare the characteristics of E. coli recovered from 7 different integrated broiler operations in South Korea. Among 200 chicken meat samples, 101 were observed to be positive for E. coli. However, the prevalence varied from 37.5% to 75.0% in chicken meats from different operations, indicating variation in E. coli occurrence among the operations. Among 101 isolated E. coli from chicken meat, 59 were identified third-generation cephalosporin-resistant E. coli and recovered from 7 different operations. A high proportion of the E. coli isolates were resistant to penicillins (89.8%), quinolones (81.4%). Among 59 third-generation cephalosporin-resistant E. coli isolates, 29 showed phenotypic and genotypic characteristics of ß-lactamase-producing E. coli. Prevalence of bla gene, blaCTX-M-1, blaCTX-M-14, blaCMY-2, and blaTEM-1, were identified in 2, 4, 8, and 16 E. coli isolates respectively and only one E. coli had both genes, blaTEM-1 and blaCTX-M-1. Pulsed-field gel electrophoresis (PFGE) analysis was performed on 29 ß-lactamase-producing E. coli isolates. In PFGE, E. coli included 7 PFGE patterns showing the same operation and an accorded both resistance to ß-lactam antibiotics and presence of the bla-gene. Our findings suggest that E. coli with resistance to third-generation cephalosporins can now be found in association with integrated broiler operations, providing the data to support the development of monitoring and preventing program in integrated operations.

Prevalence and characterization of Salmonella in two integrated broiler operations in Korea.

BACKGROUND: Vertical integration of the broiler industry allows producers to combine different biosecurity and sanitation practices, housing technologies, and feeding regimens to improve food safety. The purpose of this study was to investigate the prevalence and distribution of Salmonella, to determine the source of Salmonella contaminants, and to reveal the relationships between isolates at each step in the vertically integrated broiler production system in two representative integrated broiler companies in Korea. RESULTS: A total of 2148 samples were collected from 2 broiler breeder hatcheries, 14 broiler breeder farms, 3 broiler hatcheries, 16 broiler farms, 8 broiler transporting trucks and 6 slaughterhouses belonging to representative integrated broiler companies, and 205 (9.5%) of these samples were positive for Salmonella. The Salmonella prevalence in broiler hatcheries (34.0%) and broiler transporting trucks (62.5%) was higher (P < 0.05) whereas that in the broiler breeder hatchery (0.8%) was lower (P < 0.05), than the overall prevalence. Nine and 13 different Salmonella serotypes were isolated from integrated companies A and B, and the predominant serotypes were S. Virchow (39.7%) and S. Hadar (59.2%), respectively. Pulsed field gel electrophoresis patterns of isolates from the two operations showed significant genetic relatedness within a single system. CONCLUSIONS: In a comparison of the two operations that participated in this study, the prevalence of Salmonella differed significantly between the broiler breeder hatchery, and broiler hatcheries and broiler farms.