Erythromycin-resistant Enterococci Research Articles

Antimicrobial susceptibility patterns of environmental and hospital isolations of enterococci in Aydın

The aim of the present study was to evaluate antimicrobial susceptibilities of enterococci isolated from water, soil, waste, and food samples to determine resistance mechanisms and their transferability, and to test clonality of these environmental enterococci. The results were compared with the susceptibility patterns of hospital enterococci reported in a previous study. Samples from 9 environmental sources were cultured, and species were identified using 16S rRNA sequencing. Antimicrobial susceptibilities were determined by the agar dilution method. The minimum inhibitory concentrations were tested for vancomycin, tetracycline, teicoplanin, ampicillin, gentamicin, erythromycin, rifampicin, clindamycin, chloramphenicol, and ciprofloxacin. Resistance genes and transposons were determined by polymerase chain reaction. The transferability of resistance was tested by conjugation. A total of 57 enterococci were isolated. All erythromycin-resistant enterococci had ermB and all tetracycline-resistant isolates had tetM genes. Macrolide and tetracycline resistances were transferable by conjugation to Enterococcus faecalis JH2-2. Transconjugants were confirmed by pulsed-field gel electrophoresis analysis. Our study showed the existence of antimicrobial resistance among bacteria isolated from environmental samples. All isolates were resistant to at least one antibiotic tested. Transferability of resistance genes in environmental enterococci showed the importance of environmental resistance for public health and the potential of resistance gene dissemination. Although the sample collection times of environmental and hospital isolates were different, the possibility exists that the difference in susceptibility patterns may be the result of differences in antimicrobial pressure in hospitals and in the environment, and this should be studied further.

Insects in confined swine operations carry a large antibiotic resistant and potentially virulent enterococcal community.

BackgroundExtensive use of antibiotics as growth promoters in the livestock industry constitutes strong selection pressure for evolution and selection of antibiotic resistant bacterial strains. Unfortunately, the microbial ecology and spread of these bacteria in the agricultural, urban, and suburban environments are poorly understood. Insects such as house flies (Musca domestica) and German cockroaches (Blattella germanica) can move freely between animal waste and food and may play a significant role in the dissemination of antibiotic resistant bacteria within and between animal production farms and from farms to residential settings.ResultsEnterococci from the digestive tract of house flies (n = 162), and feces of German cockroaches (n = 83) and pigs (n = 119), collected from two commercial swine farms were isolated, quantified, identified, and screened for antibiotic resistance and virulence. The majority of samples (93.7%) were positive for enterococci with concentrations 4.2 ± 0.7 × 104 CFU/house fly, 5.5 ± 1.1 × 106 CFU/g of cockroach feces, and 3.2 ± 0.8 × 105 CFU/g of pig feces. Among all the identified isolates (n = 639) Enterococcus faecalis was the most common (55.5%), followed by E. hirae (24.9%), E. faecium (12.8%), and E. casseliflavus (6.7%). E. faecalis was most prevalent in house flies and cockroaches, and E. hirae was most common in pig feces. Our data showed that multi-drug (mainly tetracycline and erythromycin) resistant enterococci were common from all three sources and frequently carried antibiotic resistance genes including tet(M) and erm(B) and Tn916/1545 transposon family. E. faecalis frequently harbored virulence factors gelE, esp, and asa1. PFGE analysis of selected E. faecalis and E. faecium isolates demonstrated that cockroaches and house flies shared some of the same enterococcal clones that were detected in the swine manure indicating that insects acquired enterococci from swine manure.ConclusionsThis study shows that house flies and German cockroaches in the confined swine production environment likely serve as vectors and/or reservoirs of antibiotic resistant and potentially virulent enterococci and consequently may play an important role in animal and public health.

Vancomycin‐ and erythromycin‐resistant enterococci in a pig farm and its environment

A high prevalence of vancomycin- and erythromycin-resistant enterococci (VRE and ERE respectively) in a pig farm and its environment was observed. A similar structure and composition of enterococcal populations was detected between urban sewage and those associated with the pig environment. Enterococcus faecium was the most predominant species among VRE isolates from both animal and human origin. The high population similarity index (Sp) obtained comparing VRE and ERE isolates from urban sewage and pig slurry suggests that there are certain strains circulating through the food chain from farms to humans. Erythromycin resistance was present in a wider variety of clones and species of enterococci in both pigs and humans than vancomycin resistance.

Antimicrobial susceptibility patterns of competitive exclusion bacteria applied to newly hatched chickens

Competitive exclusion (CE) products are mixtures of obligate and facultative anaerobic bacteria applied to poultry hatchlings for prevention of Salmonella colonization. These mixtures have the potential to introduce bacteria with undesirable antimicrobial drug resistance traits into the human food supply. Antimicrobial drug susceptibilities of 27 obligate and facultative anaerobes isolated from a commercial CE product were evaluated with a microdilution minimal inhibitory concentration (MIC) assay. Bacteroides distasonis and Bacteroides fragilis isolates were resistant to tetracycline and other antimicrobial drugs. An Escherichia coli isolate was resistant to four antimicrobial drugs: erythromycin, penicillin, vancomycin, and tylosin. Erythromycin-resistant enterococci and vancomycin-resistant Lactococcus lactis isolates in the CE product were detected. These findings suggest that more work needs to be done to assess the potential effects of CE product use in poultry on the food supply.

Distribution and persistence of fecal bacterial populations in liquid and dewatered sludge from a biological treatment plant

The changes in composition and structure of fecal coliforms (FC) and enterococci (ENT) populations, as well as the elimination of spores of sulphite-reducing bacteria (SRB), were compared between municipal sewage and their derived sludge in a biological treatment plant in order to determine any selective reduction or adsorption to sludge during the treatment process. Additionally, the persistence of antibiotic-resistant enterococcal populations in two kinds of sludge was also considered to evaluate their potential elimination in the treatment process. Microbial indicators, vancomycin-resistant and erythromycin-resistant enterococci were enumerated. The structure and composition of FC and ENT populations were determined by biochemical fingerprinting and clustering analyses. Raw and treated sewage showed a concentration of FC 1 log unit higher than ENT and nearly 2 log units higher than spores of SRB. However, the three studied indicators showed similar concentrations in both types of sludge. Consequently, FC were eliminated in higher proportion than ENT and spores of SRB in sludge. FC and ENT populations showed high diversity and similarity population indexes for all kinds of samples. Antibiotic-resistant enterococci persisted in a similar proportion in respect to total enterococci not only in treated sewage but also in sludge. The persistence of antibiotic-resistant strains in sludge as well as in treated sewage should be considered if they are used for land disposal or for water reutilization, respectively.

Effects of tylosin use on erythromycin resistance in enterococci isolated from swine.

The effect of tylosin on erythromycin-resistant enterococci was examined on three farms; farm A used tylosin for growth promotion, farm B used tylosin for treatment of disease, and farm C did not use tylosin for either growth promotion or disease treatment. A total of 1,187 enterococci were isolated from gestation, farrowing, suckling, nursery, and finishing swine from the farms. From a subset of those isolates (n = 662), 59% (124 out of 208), 28% (80 out of 281), and 2% (4 out of 170) were resistant to erythromycin (MIC >/= 8 microg/ml) from farms A, B, and C, respectively. PCR analysis and Southern blotting revealed that 95% (65 out of 68) of isolates chosen from all three farms for further study were positive for ermB, but all were negative for ermA and ermC. By using Southern blotting, ermB was localized to the chromosome in 56 of the isolates while 9 isolates from farms A and B contained ermB on two similar-sized plasmid bands (12 to 16 kb). Pulsed-field gel electrophoresis revealed that the isolates were genetically diverse and represented a heterogeneous population of enterococci. This study suggests that although there was resistance to a greater number of enterococcal isolates on a farm where tylosin was used as a growth promotant, resistant enterococci also existed on a farm where no antimicrobial agents were used.

The composition and persistence of faecal coliforms and enterococcal populations in sewage treatment plants.

The changes in structure and composition of faecal coliforms and enterococcal populations in sewage from different treatment plants, and the elimination of vancomycin- and erythromycin-resistant enterococci (VRE and ERE, respectively) in these treatment plants was analysed to determine any selective reduction. Faecal coliforms, enterococci, VRE, ERE and spores of sulphite-reducing bacteria were enumerated using standard methods. Samples were enriched where necessary in order to isolate antibiotic resistant strains. The structure and composition of these bacterial populations were determined by biochemical fingerprinting and clustering analysis. High diversity and similarity indexes were detected among all the bacterial populations in raw and treated sewage, independently of their origin and the treatment processes employed. Antibiotic resistant strains were detected in all sewage tested and no selective reduction was observed. The faecal coliforms and enterococci populations did not differ in the sewage samples studied. The vancomycin and erythromycin resistances of the enterococcal populations were similar in the sewage samples. Resistance to both antibiotics persisted after the treatment process independently of raw sewage flow, faecal origin or size of the human population contributing to sewage. However, sewage of mixed origin (human and animal) presented a lower similarity index for the two bacterial populations compared with that of the other human sewage analysed. Although a significant reduction in bacterial populations was observed, the persistence of VRE and ERE strains in the same proportions in sewage suggests that there is no selective elimination of bacterial populations during the treatment processes. The ability of antibiotic resistance strains to survive sewage treatment systems should be considered in certain water reuse programmes.

Detection and characterization of erythromycin-resistant methylase genes in Gram-positive bacteria isolated from poultry litter.

The epidemiology of four erythromycin-resistant methylase ( erm) genes, ermA, ermB, ermC and msrA, was determined in erythromycin-resistant staphylococci, enterococci and streptococci isolated from poultry litter. All isolates were resistant to multiple antibiotics. Southern hybridization indicated that 4 of the 20 staphylococci contained the ermC gene on plasmids: on a 2.2 kb plasmid in Staphylococcus hominis and S. sciuri, on a 6.0 kb plasmid in S. xylosus, and on a 7.0 kb plasmid in S. lentus. In 16 of the 20 staphylococci, the ermA gene was harbored exclusively on the chromosome, as a double chromosomal insert on 8.0 and 6.2 kb EcoRI fragments. None of the staphylococci harbored the msrA gene. Dot-blot analysis indicated that all enterococci and streptococci hybridized with a biotinylated ermB gene probe. Southern hybridization indicated that only 2 of the 19 erythromycin-resistant enterococci contained the ermB gene on plasmids. The gene was localized on 4.0 kb and 5.9 kb plasmids, respectively, in two Enterococcus faecium isolates. Results from our studies indicate that the patterns of occurrence of erm genes, the sizes of the plasmids and the copy numbers of the inserts were different from the existing information on the presence of erm genes in clinical strains of Staphylococcus spp.

Effect of tylosin used as a growth promoter on the occurrence of macrolide-resistant enterococci and staphylococci in pigs.

This study was conducted to investigate the effect of tylosin used as a growth promoter on the occurrence of macrolide-resistant enterococci and staphylococci in pigs. Two identical feeding experiments with 10 pigs each were conducted to evaluate the effect of tylosin on the occurrence of erythromycin resistance among enterococci sampled from feces and Staphylococcus hyicus sampled by skin swabs. Half of the pigs were fed antibiotic-free feed and the other half feed with tylosin (30 microg/g) added. For each pig, weekly fecal samples and skin swabs were collected and examined for erythromycin-susceptible and erythromycin-resistant enterococci and S. hyicus, respectively. There was an immediate effect of tylosin on the fraction of resistant enterococci recovered, increasing by a factor 2.4; the effect on S. hyicus was more gradual, increasing at a rate of about 8% per day and totaling a five-fold increase over 20 days. Thus, a clear effect on resistance occurrence was demonstrated, not only in intestinal bacteria, but also on bacteria sampled from the skin.

Characteristics of Tn5307 exchange and intergeneric transfer of genes associated with nisin production

Transfer of the Lactococcus lactis 11454 nisin-sucrose conjugative transposon, Tn5307, was investigated to develop a methodology for conjugation of this element to other lactic acid bacteria. Tn5307 exchange was sensitive to temperature and pH but was not affected by protease or amylase treatments to donor cells. Moreover, conjugation studies demonstrated that the direct-plate method could be employed to rapidly identify LM2301 transconjugants able to transfer Tn5307 at least ten times more efficiently than 11454. Intergeneric transfer of nisin and sucrose genes between L. lactis and a dairy Enterococcus sp. was also investigated. Erythromycin-resistant Enterococcus sp. recipients were developed by electro-transformation with pGK13 or by conjugal introduction of the broad-host-range plasmid pAM beta 1. Matings between L. lactis 11454 and an Enterococcus sp. recipient that contained pAM beta 1 yielded sucrose-positive, nisin-immune transconjugants at a frequency of 2.3 x 10(-9) transconjugants per donor cfu. Agar-overlay assays for nisin production revealed that enterococcal transconjugants did not produce the bacteriocin, but DNA.DNA hybridization with a nisA-specific probe demonstrated that these bacteria had acquired the nisin structural gene.

Occurrence of glycopeptide resistance among Enterococcus faecium isolates from conventional and ecological poultry farms.

Occurrence of glycopeptide resistance among Enterococcus faecium isolates from conventional and ecological poultry farms.