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This paper presents the findings of a large-scale study on antibiotic resistance in bacteria found in farm animal feces across Russia. The study included 6578 samples of farm animal manure from 13 regions in Russia, with the help of citizen scientists. Molecular and microbiological methods were used to analyze 1111 samples of E. coli. The microbiological analysis focused on culturing the microorganisms present in the fecal samples on selective media for E. coli and evaluating the sensitivity of the bacteria to different antibiotics, including ampicillin, tetracycline, chloramphenicol, cefotaxime, and ciprofloxacin. The molecular analysis involved isolating the genomic DNA of the bacteria and conducting PCR assays to detect the vanA, vanB, and mcr-1 antibiotic resistance genes. The results demonstrated significant differences in antibiotic sensitivity of the samples that are morphologically identical to E. coli from different regions. For example, 98.0% and 82.5% of E. coli and other fecal bacterial isolates from the Omsk and Vologda regions lacked antibiotic resistance genes, while 97.7% of samples from the Voronezh region possessed three resistance genes simultaneously. The phenotypic antibiotic sensitivity test also revealed regional differences. For instance, 98.1% of fecal bacterial samples from cattle in the Udmurt Republic were sensitive to all five antibiotics tested, whereas 92.8% of samples from the Voronezh region showed resistance to all five antibiotics. The high level of antibiotic resistance observed may be attributed to their use in farming practices. The distinctive feature of our research is that comprehensive geographical coverage was achieved by using a citizen science platform. Citizen scientists, specifically students from colleges and universities, were responsible for the collection and initial analysis of samples. The project attracted 3096 student participants, enabling the collection and analysis of a significant number of samples from various locations in Russia. [ABSTRACT FROM AUTHOR]

AIM: The study aimed to investigate vanA/vanB resistance genes and to determine the frequency of vancomycin-variable enterococci(VVE). BACKGROUND: Enterococci that are phenotypically-susceptible to vancomycin but harbor glycopeptide resistance genes are called VVE. These strains cannot be detected by phenotypic methods, but it is possible by molecular methods. METHODS: Enterococcal strains, sensitive to vancomycin, were included in the study. These isolates were analyzed for the vanA and vanB genes by both monoplex and multiplex conventional PCR. ''Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR)'' method was used to investigate the clonal relationship of the isolates. RESULTS: The study included 158(62.94%) E.faecalis and 93(37.05%) E.faecium strains that were susceptible to vancomycin.According to the results of multiplex PCR using vanA primers and vanB primers together, the vanA gene was detected in one(0.39%) E.faecalis isolate; the vanB gene was detected in two E.faecium and 98 E.faecalis isolates (39.84%)(Figure 1).According to the results of monoplex PCR using vanA primers, the vanA gene was detected in one (0.39%) E.faecalis isolate (Figure 2); As a result of monoplex PCR performed with vanB primers, the vanB gene was not found in any sample (Figure 3).No dominant pattern was observed by ERIC-PCR(Figure 4). CONCLUSION: When vanA and vanB primers were investigated together with multiplex PCR, it was observed that the vanB primer caused false positivity in agarose gel electrophoresis because it formed a band close to the base pair.Although enterococci with vanA and vanB genes were detected at a very low prevalence as a result of our study, the detection of VVE isolates is essential to prevent antimicrobial resistance.

The study screened the occurrence of VanA,VanB,Vanc genes in Staph aureus vancomycin-resistant isolates recovered in Hilla -hospitals. From various clinical samples, 160 isolates from patients between July and November of 2022 were included in the search. Among these samples are (65) burn samples, (55) urine samples, and (40) samples from Blood. 40 (or 25%) of the 160 isolates were Staph aureus. Characterization of the VanA, VanB, and VanC genes in Staph aureus isolates obtained from various specimens is the main goal of the work. The RESULTS of samples were obtained from a variety of clinical specimens, including 20 (40.625%) burn swabs, 10 (25%) urine samples, and 10 (34.375%) blood. Staph aureus was resistant to Cefepime (91.7%), Piperacillin (73.3%), and Ceftazidime (71.7%), although little relationship was detected with Aztreonam resistance (35%). 40 isolates were used for antibiotic sensitivity test (AST). Only 18.3% and 15% of isolates, respectively, were resistant to meropenem and imipenem. Fluoroquinolone resistance to lomefloxacin, ciprofloxacin, ofloxacin, and levofloxacin was 51.7%, 48.3%, 35%), and 30%, respectively. While the most prevalent vancomycin resistance genes were found in 40 isolates of S. aureus and were discovered by VanC 40 (100%), VanA 20 (100%), and VanB28 (70%), according to the current investigation.In Conclusion :The PCR-reaction method was discovered to be an easy-to- use tool for detecting staph aureus by pcr that can be regularly used for identifying staph aureus and detection the molecular resistance apparatuses to vancomycin in staph aureus by VanA, VanB, VanC), efflux systems plays To choose the best antibiotics, prevent the establishment of resistant strains, and create novel antimicrobial. medications, it is beneficial to research the particular resistance mechanisms vancomycin to staph aureus. [ABSTRACT FROM AUTHOR]

Objectives: Enterococcus faecalis is a gram positive diplococci, highly versatile and a normal commensal of the gut microbiome. Resistance to vancomycin is a serious issue in various health-care setting exhibited by vancomycin resistant Enterococci (VRE) due to the alteration in the peptidoglycan synthesis pathway. This study is thus aimed to detect the VRE from the patients with root caries from the clinical isolates of E. faecalis and to evaluate the in-silico interactions between vanA and the Aegles marmelos bio-compounds.Methods : : E. faecalis was phenotypically characterized from 20 root caries samples and the frequency of vanA and vanB genes was detected by polymerase chain reaction (PCR). Further crude methanolic extracts from the dried leaves of A. marmelos was assessed for its antimicrobial activity. This is followed by the selection of five A. marmelos bio-compounds for the computational approach towards the drug ligand interactions.Results : : 12 strains (60%) of E. faecalis was identified from the root caries samples and vanA was detected from two strains (16%). Both the stains showed the presence of vanA and none of the strains possessed vanB. Crude extract of A. marmelos showed promising antibacterial activity against the VRE strains. In-silico analysis of the A. marmelos biocompounds revealed Imperatonin as the best compound with high docking energy (–8.11) and hydrogen bonds with < 140 TPSA (Topological polar surface area) and zero violations.Conclusion : : The present study records the VRE strains among the root caries with imperatorin from A. marmelos as a promising drug candidate. However the study requires further experimentation and validation.

Abstract Background Vancomycin-resistant Enterococci (VRE) represent a critical medical and public health concerns due to their association with serious nosocomial infections and a high risk of mortality. We aimed to reveal the pooled prevalence of VRE and antimicrobial resistance profiles among enterococci clinical isolates in Egypt. Methods A PubMed, Scopus, Google Scholar, and Web of Science literature search was carried out in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. Only published studies documenting the prevalence of VRE between 2010 and 2022 were included. Using the random effects model and the 95% confidence intervals, the pooled estimate of VRE was calculated by MedCalc Version 20.113. Cochran’s Q and I 2 tests were used to evaluate the degree of heterogeneity, and publication bias was examined by visually examining the funnel plot and its associated tests (Begg’s and Egger’s tests). Results The pooled prevalence of VRE among enterococci clinical isolates in Egypt was estimated to be 26% (95% CI 16.9 to 36.3). E. faecalis had a greater pooled prevalence than E. faecium, with 61.22% (95% CI 53.65 to 68.53) and 32.47% (95% CI 27 to 38.2), respectively. The VanA gene is more frequent than the VanB gene among VRE, with a pooled prevalence of 63.3% (95% CI 52.1 to 73.7) and 17.95% (95% CI 7.8 to 31), respectively. The pooled resistance rate of linezolid was substantially lower than that of ampicillin and high-level gentamicin (HLG) 5.54% (95% CI 2.33 to 10%), 65.7% (95% CI 50.8 to 79.2%), and 61.1% (95% CI 47.4 to 73.9), respectively. Conclusion The prevalence of VRE is alarmingly high in Egypt. It is imperative that antimicrobial stewardship activities and infection control programs are strictly adhered to and implemented to prevent further escalation of the problem.

Purpose: Methicillin-resistant Staphylococcus aureus, a common bacterial pathogen causes various infections. The acquisition of various antimicrobial-resistant genes in S. aureus has led to the transformation of this bacterium into a superbug. Vancomycin resistance among MRSA isolates is an emerging threat in empirical therapy of various infections. The study was hence aimed to find out the susceptibility status of S. aureus isolates toward vancomycin and detect mecA, vanA, and vanB genes among the isolates. Methods: A total of 1245 clinical samples from the participants attending a tertiary care hospital in Kathmandu were processed. S. aureus isolated from the samples were subjected to antibiotic susceptibility patterns using the modified Kirby-Bauer disk diffusion method. Agar dilution method was used to determine the minimum inhibitory concentration of vancomycin. The antibiotic-resistant genes such as mecA, vanA, and vanB among S. aureus isolates were screened by a conventional polymerase chain reaction. Results: Of 1245 samples, 80 S. aureus were identified. Out of which, 47.5% (38/80) were phenotypically confirmed MRSA isolates. mec A gene was detected in 84.2% (32/38) of MRSA isolates. 10.5% (4/38) were confirmed as vancomycin-intermediate S. aureus (VISA) by MIC determination. None of the isolates was positive for the vanA gene; however, 2 isolates were found to possess the vanB gene. The 2 isolates have vancomycin MIC breakpoints of 4 to 8 μg/mL. Conclusion: There might be a spreading of vancomycin resistance among S. aureus, creating serious public health problems. Therefore, measures to limit vancomycin resistance should be considered in healthcare facilities as immediately as possible. [ABSTRACT FROM AUTHOR]

Background and Objective: Enterococci are typically found in a healthy human gastrointestinal tract but can cause severe infections in immunocompromised patients. Such infections are treated with antibiotics. This study addresses the rising concern of antimicrobial resistance (AMR) in Enterococci, focusing on the prevalence of vancomycin-resistant enterococcus (VRE) strains. Materials and Methods: The pilot study involved 140 Enterococci isolates collected between 2021 and 2022 from two multidisciplinary hospitals (with and without local therapeutic drug monitoring protocol of vancomycin) in Latvia. Microbiological assays and whole genome sequencing were used. AMR gene prevalence with resistance profiles were determined and the genetic relationship and outbreak evaluation were made by applying core genome multi-locus sequence typing (cgMLST). Results: The acquired genes and mutations were responsible for resistance against 10 antimicrobial classes, including 25.0% of isolates expressing resistance to vancomycin, predominantly of the vanB type. Genetic diversity among E. faecalis and E. faecium isolates was observed and seven potential outbreak clusters were identified, three of them containing sequence types ST6, ST78 and ST80. The prevalence of vancomycin resistance was highest in the hospital without a therapeutic drug-monitoring protocol and in E. faecium. Notably, a case of linezolid resistance due to a mutation was documented. Conclusions: The study illustrates the concerning prevalence of multidrug-resistant Enterococci in Latvian hospitals, showcasing the rather widespread occurrence of vancomycin-resistant strains. This highlights the urgency of implementing efficient infection control mechanisms and the need for continuous VRE surveillance in Latvia to define the scope and pattern of the problem, influencing clinical decision making and planning further preventative measures.

Background: Vancomycin-resistant Enterococci (VRE) represent a critical medical and public health concerns due to their association with serious nosocomial infections and a high risk of mortality. We aimed to reveal the pooled prevalence of VRE and antimicrobial resistance profiles among enterococci clinical isolates in Egypt. Methods: A PubMed, Scopus, Google Scholar, and Web of Science literature search was carried out in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. Only published studies documenting the prevalence of VRE between 2010 and 2022 were included. Using the random effects model and the 95% confidence intervals, the pooled estimate of VRE was calculated by MedCalc Version 20.113. Cochran's Q and I2 tests were used to evaluate the degree of heterogeneity, and publication bias was examined by visually examining the funnel plot and its associated tests (Begg's and Egger's tests). Results: The pooled prevalence of VRE among enterococci clinical isolates in Egypt was estimated to be 26% (95% CI 16.9 to 36.3). E. faecalis had a greater pooled prevalence than E. faecium, with 61.22% (95% CI 53.65 to 68.53) and 32.47% (95% CI 27 to 38.2), respectively. The VanA gene is more frequent than the VanB gene among VRE, with a pooled prevalence of 63.3% (95% CI 52.1 to 73.7) and 17.95% (95% CI 7.8 to 31), respectively. The pooled resistance rate of linezolid was substantially lower than that of ampicillin and high-level gentamicin (HLG) 5.54% (95% CI 2.33 to 10%), 65.7% (95% CI 50.8 to 79.2%), and 61.1% (95% CI 47.4 to 73.9), respectively. Conclusion: The prevalence of VRE is alarmingly high in Egypt. It is imperative that antimicrobial stewardship activities and infection control programs are strictly adhered to and implemented to prevent further escalation of the problem. [ABSTRACT FROM AUTHOR]

ABSTRACT: Objectives: The aim of this study was to evaluate the occurrence of glycopeptide resistance in enterococci and coagulase-negative staphylococci (CoNS) and to determine the susceptibilities of the identified glycopeptide-resistant isolates to dalbavancin. Methods: Twenty-two medical laboratories participated in the study conducted in 2016/17 by the Paul-Ehrlich-Society for Chemotherapy. Each laboratory was asked to collect 30 Enterococcus spp. (limited to Enterococcus faecalis and Enterococcus faecium) and 30 CoNS isolates consecutively from hospitalised patients with a proven or suspected infection. Results: A total of 1285 isolates were collected, comprising 364 E. faecalis, 291 E. faecium and 630 CoNS. No E. faecalis isolates (0%) but 76 E. faecium isolates (26.1%) were vancomycin-resistant, of which 21 showed the VanA type and 55 the VanB type. The proportion of vancomycin-resistant strains among E. faecium isolates from patients in intensive care units (21.6%) was significantly lower than that from patients on regular wards (30.5%). Among the CoNS, 67 isolates (10.6%) were teicoplanin-resistant but none were vancomycin-resistant, with resistance only detected in Staphylococcus epidermidis (12.2%), Staphylococcus haemolyticus (17.9%) and Staphylococcus hominis (13.2%). Dalbavancin at ≤0.25 mg/L inhibited all VanB-type enterococci and 95.5% of teicoplanin-resistant CoNS. Conclusion: The level of glycopeptide resistance in E. faecalis remains very low in Germany but achieved 26% in E. faecium and was >10% in CoNS. Dalbavancin appears to be a feasible option for treating infections caused by VanB-type vancomycin-resistant E. faecium and teicoplanin-resistant CoNS.

Morbidity and mortality associated with antimicrobial resistance is a global public health problem. Vancomycinresistant Enterococcus faecium, which is frequently seen in healthcare-associated infections, is a Gram-positive cocci that the he World Health Organization has given high priority on its list of priority pathogens with antimicrobial resistance. Recently, the rate of vancomycin-resistant Enterococcus faecium isolation in invasive infections has been reported as 15.8% in Turkey. The main mechanism of resistance to vancomycin is alteration of the D-alanyl-D-alanine terminal molecule in the target region, and decreased affinity for the antibiotic. The van gene responsible for vancomycin resistance is either chromosomally encoded or acquired by mobile genetic elements such as plasmid and transposon. The main mechanism of its spread is that mobile genetic elements can be transferred within the same species or between different bacteria like Staphylococcus aereus. In this review, data, extracted by PubMed and Web of Science Databases searching with the terms "Vancomycin-resistant Enterococcus faecium", "Vancomycin resistance mechanisms", "vancomycin resistance in Gram-positive cocci" is summarized and briefly discussed with recommendations. [ABSTRACT FROM AUTHOR]

Copyright of Kocatepe Veterinary Journal / Kocatepe Veteriner Dergisi is the property of Afyon Kocatepe University, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Detrimental consequences of antibiotic treatment may include long-lasting disruption of the gut microbiota. Previous studies found no negative effects of antibiotics on metabolic health, although individualized responses were observed. Here, we aimed to investigate the subject-specific response to vancomycin use in tissue-specific insulin sensitivity by stratifying individuals based on the presence of antibiotic resistance genes (ARGs) or opportunistic pathogens (OPs) in the baseline fecal microbiota. Quantitative Polymerase Chain Reaction (qPCR) was used to detect ARGs and OPs in DNA isolated from fecal samples of 56 males with overweight/obesity (Body Mass Index: 25–35 kg/m2) and impaired glucose metabolism (fasting plasma glucose ≥5.6 mmol/L and/or 2-hour glucose 7.8–11.1 mmol/L). A two-step hyperinsulinemic-euglycemic clamp was performed to determine tissue-specific insulin sensitivity. Abdominal subcutaneous adipose tissue (AT) gene expression was assessed using Affymetrix microarray. Gut microbial composition was determined using the Human Intestinal Tract Chip (HITChip) microarray. At baseline, the vancomycin resistance gene vanB was present in 60% of our population. In individuals that were vanB-negative at baseline, AT insulin sensitivity (insulin-mediated suppression of plasma free fatty acids) improved during vancomycin use, while it decreased among vanB-positive individuals (% change post versus baseline: 14.1 ± 5.6 vs. −6.7 ± 7.5% (p = .042)). The vancomycin-induced increase in AT insulin sensitivity was accompanied by downregulation of inflammatory pathways and enrichment of extracellular matrix remodeling pathways in AT. In the vanB-positive group, well-known vanB-carrying bacteria, Enterococcus and Streptococcus, expanded in the gut microbiome. In conclusion, microbiome composition and adipose tissue biology were differentially affected by vancomycin treatment based on fecal vanB carriage.

Antibiotic resistance has become a major threat to human health around the world, but its spread through the aquatic environment has been often overlooked. This study aimed to determine the occurrence of vancomycin-resistant Staphylococcus aureus in hospital wastewaters and their transmission into public water bodies in Kerala, India. A total of 113 S. aureus were isolated from three hospital effluents in Kerala, India. Standard disc diffusion and the strip method were used for antibiotic susceptibility testing and minimum inhibitory concentration detection. Plasmid-mediated vancomycin resistance was confirmed by plasmid curing and conjugation; resistant genes were detected by the polymerase chain reaction (PCR). Nearly 76% of S. aureus isolates were resistant to β-lactams, chloramphenicol, macrolides, aminoglycosides, and glycopeptide class of antibiotics. Among the vancomycin-resistant Staphylococcus aureus (VRSA) isolates, the prevalence rates of vanA and vanB resistance-encoding genes were 46.5 and 59.3%, respectively. Through the broth mating method, vanA gene was successfully transferred from VRSA donor to vancomycin-sensitive S. aureus. The study strongly indicates the contamination of water bodies with antibiotic-resistant bacteria from hospital discharges, their dissemination and possible transfer to microbes in the aquatic environment, posing a serious threat for public health. HIGHLIGHTS High vancomycin resistance was observed in S. aureus isolates in the hospital effluent.; Vancomycin-resistant Staphylococcus aureus (VRSA) isolates with vanA and vanB resistance-encoding genes have a higher chance of surviving in the sewage treatment plants.; Horizontal transfer of the resistance gene was confirmed by conjugation.; The VRSA isolates have a strong capacity to acquire or transfer antibiotic-resistant genes, posing a threat to public health.;

Copyright of Journal of Basrah Researches (Sciences) is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

COVID-19 offers ideal premises for bacteria to develop antimicrobial resistance. In this study, we evaluated the presence of several antimicrobial resistance genes (ARG) in vancomycin-resistant Enterococcus (VRE) isolated from rectal swabs from patients at a hospital in Cluj-Napoca, Romania. Rectal swabs were cultivated on CHROMID® VRE (bioMérieux, Marcy—l' Étoile, France) and positive isolates were identified using MALDI-TOF Mass Spectrometry (Bruker Daltonics, Bremen, Germany) and further analyzed using the PCR technique for the presence of the following ARGs: van A, van B, tet(M), tet(L), ermB, msrA, mefA, aac(6′)-Im, aph(2)-Ib, ant(4′)-Ia, sul1, sul2, sul3, and NDM1. We isolated and identified 68 isolates of Enterococcus faecium and 11 isolates of Enterococcus faecalis. The molecular analysis showed 66 isolates positive for the vanA gene and eight positive for vanB. The most frequent association of ARG in VRE was vanA-tet(M)-ermB. There was no statistically significant difference between Enterococcus faecium and Enterococcus faecalis regarding ARGs. Our work proves that during the COVID-19 pandemic, highly resistant isolates of Enterococcus were present in patients in the intensive care unit; thus, better healthcare policies should be implemented for the management and control of these highly resistant isolates in the future. [ABSTRACT FROM AUTHOR]

Background & objectives: Infections caused by vancomycin-resistant Enterococci are difficult to treat given the limited therapeutic alternatives. Different gene clusters are known to confer vancomycin resistance. vanA and vanB genes are transferable and are clinically relevant. This cross-sectional study aimed to identify the vancomycin-resistant genotypes in the strains causing urinary tract infection and also to test the in vitro efficacy of linezolid and pristinamycin against the vancomycin-resistant isolates. Methods: Antimicrobial resistance profile of 118 enterococcal isolates was evaluated. Minimum inhibitory concentration of vancomycin, teicoplanin and high-level gentamicin (HLG) was determined by micro broth dilution. The vancomycin-resistant isolates were tested against linezolid and pristinamycin by micro-broth dilution and E strip method. The presence of vancomycin-resistant genes was detected by multiplex polymerase chain reaction and was sequenced and analyzed. Results: Most commonly isolated species were Enterococcus faecalis (76.9%) and Enterococcus faecium (16.9%). It was found that 43 per cent of the isolates were resistant to HLG and 16.9 per cent to vancomycin. Higher resistance was seen against fluoroquinolones, erythromycin, tetracycline and β-lactam drugs. However, 5.08 per cent strains were resistant to tigecycline. All vancomycin-resistant strains were sensitive to pristinamycin and one was resistant to linezolid. vanA and vanB gene were found in 15 and five isolates, respectively. The gene sequences were submitted to NCBI gene bank and accession numbers were obtained. Interpretation & conclusions: The present study showed prevalence of vanA and vanB genes carrying Enterococcus in a tertiary care centre in north India. The emergence of resistance against drugs such as tigecycline and linezolid is a topic of concern as it will be a therapeutic challenge for physicians. [ABSTRACT FROM AUTHOR]

Background/Purpose: To evaluate the ability of quadruple Taqman probe real-time PCR to the detection of vanA, vanB and vanM in enterococcal isolates. Methods: A total of 343 strains, including 253 vancomycin-resistant enterococcus (VRE) strains and 90 non-VRE strains, were tested by both quadruple Taqman probe real-time PCR and gel-based PCR assay. Results: When differentiating among three genotypes of vanA, vanB and vanM in VRE strains, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), diagnostic accuracy and consistency of the quadruple Taqman probe real-time PCR were all 100%. Minimum.Inhibitory concentration (MIC) results showed that there was a wide MIC range of vancomycin and teicoplanin for the strains that harboring vanA/vanM gene respectively or harboring vanA and vanM genes simultaneously. However, the VRE strains with vanB genotype all were sensitive to teicoplanin. Conclusion: Considering the excellent PPV and low NPV, real-time PCR would be useful to monitor VRE-colonized or infected patients. However, further evaluation of the assay's performance in the clinical specimens is required, especially when considering that high level of PCR inhibitors present in these samples.

Abstract Background Currently, hospitals have been forced to divert substantial resources to cope with the ongoing coronavirus disease 2019 (COVID-19) pandemic. It is unclear if this situation will affect long-standing infection prevention practices and impact on healthcare associated infections. Here, we report a nosocomial cluster of vancomycin-resistant enterococci (VRE) that occurred on a COVID-19 dedicated intensive care unit (ICU) despite intensified contact precautions during the current pandemic. Whole genome sequence-based typing (WGS) was used to investigate genetic relatedness of VRE isolates collected from COVID-19 and non-COVID-19 patients during the outbreak and to compare them to environmental VRE samples. Methods Five VRE isolated from patients (three clinical and two screening samples) as well as 11 VRE and six vancomycin susceptible Enterococcus faecium (E. faecium) samples from environmental sites underwent WGS during the outbreak investigation. Isolate relatedness was determined using core genome multilocus sequence typing (cgMLST). Results WGS revealed two genotypic distinct VRE clusters with genetically closely related patient and environmental isolates. The cluster was terminated by enhanced infection control bundle strategies. Conclusions Our results illustrate the importance of continued adherence to infection prevention and control measures during the COVID-19 pandemic to prevent VRE transmission and healthcare associated infections.

Vancomycin-resistant enterococci (VRE), especially Enterococcus faecium, have been a global concern, often causing serious healthcare-associated infections. We established a rapid approach for detecting E. faecium and vancomycin-resistance genes (vanA and vanB) in clinical samples using isothermal recombinase polymerase amplification (RPA) combined with a lateral-flow (LF) strip. Specific RPA primer sets and probes for ddl (to identify the presence of E. faecium) vanA and vanB genes were designed. The RPA reaction was performed under isothermal condition at 37 °C within 20 min and read using the LF strip within a further 5 min. A total of 141 positive blood-cultures and 136 stool/rectal swab samples were tested using RPA-LF method compared to the conventional PCR method. The RPA-LF method exhibited 100% sensitivity in both blood-culture (60 E. faecium; 35 vanA type and two vanB type) and stool/rectal-swab samples (63 E. faecium and 36 vanA type) without cross-reaction (100% specificity). The lower detection limit of the RPA-LF was approximately 10 times better than that of the conventional PCR method. The RPA-LF method is an alternative rapid method with excellent sensitivity and specificity for detecting E. faecium, vanA, and vanB, and it has the potential to be used as a point-of-care device for VRE therapy and prevention. [ABSTRACT FROM AUTHOR]

Antibiotic resistance has become a major threat to human health around the world, but its spread through the aquatic environment has been often overlooked. This study aimed to determine the occurrence of vancomycin-resistant Staphylococcus aureus in hospital wastewaters and their transmission into public water bodies in Kerala, India. A total of 113 S. aureus were isolated from three hospital effluents in Kerala, India. Standard disc diffusion and the strip method were used for antibiotic susceptibility testing and minimum inhibitory concentration detection. Plasmid-mediated vancomycin resistance was confirmed by plasmid curing and conjugation; resistant genes were detected by the polymerase chain reaction (PCR). Nearly 76% of S. aureus isolates were resistant to ß-lactams, chloramphenicol, macrolides, aminoglycosides, and glycopeptide class of antibiotics. Among the vancomycin-resistant Staphylococcus aureus (VRSA) isolates, the prevalence rates of vanA and vanB resistance-encoding genes were 46.5 and 59.3%, respectively. Through the broth mating method, vanA gene was successfully transferred from VRSA donor to vancomycin-sensitive S. aureus. The study strongly indicates the contamination of water bodies with antibiotic-resistant bacteria from hospital discharges, their dissemination and possible transfer to microbes in the aquatic environment, posing a serious threat for public health. [ABSTRACT FROM AUTHOR]

Abstract Background The incidence of human infection and colonization with methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) has increased in the recent years. Environmental sources, including bird droppings, might play an important role as resistance reservoirs. Results Fresh fecal samples were collected from rooks and wild-living birds during the autumn-winter period of 2016/2017, and tested for the presence of bacteria associated with human diseases. Besides bacteria representing the genera Enterococcus, Campylobacter, Escherichia, and Staphylococcus, Enterobacter, Citrobacter, Proteus, Hafnia, and Pseudomonas were also identified. The susceptibility of S. aureus and Enterococcus spp. isolates to methicillin, and vancomycin and teicoplanin, respectively, was analyzed to assess the avian wildlife as a reservoir of MRSA and VRE strains. Twenty-two percent of all S. aureus isolates were methicillin-resistant. These strains were screened by polymerase chain reaction (PCR), using the most widely used primer sets specific for the mecA gene. Twenty percent of all Enterococcus strains were phenotypically vancomycin-resistant. The presence of van resistance genes in these strains was investigated by PCR using vanA and vanB gene-specific primers. A good correlation between mecA gene detection and disc diffusion data was observed, while some discrepancy was noted between the PCR data and the vancomycin/teicoplanin phenotypic resistance pattern. The incidence of strains resistant to methicillin and glycopeptide antibiotics in wild-living birds was twice that in rooks. Conclusions The study suggests that rooks from urban areas and passerine birds from the natural habitat carry antibiotic-resistant Enterococcus spp. and S. aureus strains, probably reflecting the presence of such isolates in the environmental food sources.

The purpose of the present study was to determine the antimicrobial susceptibility of vancomycin-resistant and Enterococcus faecium strains isolated from urine samples of dogs. A total of 22 Enterococcus sp. samples were isolated and identified from 100 urine samples collected by cystocentesis from dogs of both sexes. The identification with species specific primers for multiplex PCR revealed that all 22 isolates (100%) belonged to E. faecium. Vancomycin resistance was found in 10 (45%) samples of E. faecium strains with PCR study by vanA and vanB primers.

Background: Vancomycin-resistant enterococci (VRE), particularly Enterococcus faecium (VREfm), can cause serious nosocomial infections, and have been responsible for healthcare-associated outbreaks. Spreading of VREfm can occur both clonally and by the dissemination of mobile genetic elements.Aim: To report prospective analysis of whole-genome sequencing (WGS) data, including both core-genome multi-locus sequence typing (cgMLST) and transposon analysis, during a vanB VREfm outbreak.Methods: Screening for vanB-positive VREfm isolates was performed by real-time polymerase chain reaction (PCR) on an overnight enriched broth and, if positive, subculture was performed. vanB-positive VREfm isolates underwent WGS. Generated data were used for molecular typing that was performed by cgMLST using SeqSphere. For transposon characterization, sequence data were mapped against the reference sequence of transposon Tn1549 using CLC Genomics Workbench, or de-novo assemblies were used for BLASTN comparisons.Results: In total, 1358 real-time PCRs were performed. Two hundred and fifty-one specimens from 207 patients tested positive on PCR for vanB, of which 13 specimens obtained from six patients were identified as vanB VREfm positive on culture. These six patients harboured seven unique isolates belonging to four cluster types: CT118 (N=2), CT2483 (N=3), CT2500 (N=1) and CT2501 (N=1). Transposon analysis revealed the presence of an identical vanB-carrying transposon in the isolates cultured from all six patients that could be linked based on epidemiological data.Conclusion: A vanB VREfm outbreak occurred in the study hospital, including six patients with isolates belonging to four cluster types. In-depth transposon analysis revealed that dissemination of transposon Tn1549 rather than clonal spread was the cause of the outbreak. [ABSTRACT FROM AUTHOR]

The appearance isolates of vancomycin-resistant/intermediate S. aureus (VRSA/VISA), in the community, becomes a big concern in worldwide public health in both developed as well as developing countries. Hence, the purpose of the current research aimed to examine the molecular determinants of VRSA and VISA isolates among burn wound and UTIs patients. Out of 692 clinical specimens (non-duplicate) were collected from the main hospitals, Burn Center, and some chief clinical laboratories in Najaf City. The S. aureus isolates were recognized based on biochemical examinations, and finally confirmed by Vitek-2 compact system that 134/692(19.364%) isolates were returned to S. aureus involved 101/ 589(17.147%) and 33/103(32.038%) were obtained from urine and burn wounds sources, respectively. The data of current work was revealed out of 692 clinical specimens were recorded 571(82.514%) bacterial growth included 218(31.502%) gram-positive and 353(51.011%) was gram-negative while 121(32.537%) was no bacterial growth. The data observed the frequency of urinary tract infections were pointed out to be most in female 352(59.762%%) than male121 (20.543%). Furthermore, the rate of S. aureus isolates in females was 77(21.875%) compared with 24(19.834 %%) in males. At the same matter, the rate of burn wound infections between female and male were related were 47/103 (45.631 %%) in female while 51 (49.514%) in male. Furthermore, the rate of S. aureus isolates in female and male were 15(31.914%) and 18(35.294 %%) respectively. Vancomycin screening test for VRSA isolated was evaluated according to CLSI guide, the data revealed out of 134 S. aureus isolates were 51/ 134(38.059%) isolates able to grow at concentration 4µg/ml vancomycin. Data of the Vitek-2 system revealed among 51 isolates were 18 isolates as VRSA 2 isolates as VISA. Current work also included molecular investigation that was screened using PCR. The data revealed vanA was 6(30%), while vanB was detected only in 3(15%) isolates. This study observed vanZ at 1(5%) isolate and this result may be for first time is recoded locally. While other genes like vanS, vanH, vanY, vanR and vanX were no observed. Also, current work observed a high rate of mecA and blaZ genes among 20 isolates of VRSA/VISA reached 18 (90%) and 16 (80%), respectively. [ABSTRACT FROM AUTHOR]

Stefan Borgmann,1 Beate Rieß,1 Rabea Siegmund,1 Guido Werner,2 Ingo Klare2 1Department of Infectious Diseases and Infection Control, Ingolstadt Hospital, Ingolstadt, Germany; 2National Reference Centre for Staphylococci and Enterococci, Robert Koch Institute, Wernigerode Branch, Wernigerode, Germany Purpose: According to the WHO vancomycin-resistant Enterococcus faecium (VRE) belongs to the microorganisms for which new antibiotics are urgently needed. Depending on the type of vancomycin resistance vanA gene VRE is differentiated from vanB VRE and other types. In this retrospective analysis the results of VRE surveillance performed at a German tertiary hospital with approximately 1,200 beds between 2013 and 2017 are shown.Patients and methods: Rectal screening swabs were taken at admission and once per week on the early rehabilitation ward of Ingolstadt Hospital (ERWIN) but not at other wards. The number of VRE colonized patients was evaluated by using appropriate computer software (LabCentre, Hybase). The Hybase program was also used to find out the number of Saccharomyces boulardii and multi-susceptible Escherichia coli Nissle in blood cultures of patients at ERWIN. The mechanism of vancomycin resistance was examined by PCR and clonality of VRE strains was analyzed by pulsed-field gel electrophoresis.Results: Between 2013 and 2015 the number of VRE increased from 30 to 78 per year whereas in 2016 and 2017 the number declined to 51. Systematic analysis of the laboratory data revealed that this increase was driven by oligoclonal transmission of vanB VRE on ERWIN until August 2016 despite performing intensified infection control measures. However, afterward the number of VRE decreased at ERWIN and subsequently at the other wards. While searching for the reason behind this beneficial development we noticed that at ERWIN, patients treated with antibiotics received two probiotic medications simultaneously (S. boulardii, E. coli Nissle) for the duration of the antibiotic therapy plus an additional 2 days. There was no indication of side effects caused by these microorganisms, particularly no infections.Conclusion: Application of S. boulardii and E. coli Nissle was safe and associated with reduced transmission of VRE from patient to patient at ERWIN. Therefore, in our setting, probiotic treatment of patients receiving antibiotics contributed to the increase of patients’ safety. Keywords: microbiota, oligoclonal spread, outbreak, probiotics, vanB, bacterial spread, circuit model of bacterial transmission, vanA

COVID-19 offers ideal premises for bacteria to develop antimicrobial resistance. In this study, we evaluated the presence of several antimicrobial resistance genes (ARG) in vancomycin-resistant Enterococcus (VRE) isolated from rectal swabs from patients at a hospital in Cluj-Napoca, Romania. Rectal swabs were cultivated on CHROMID® VRE (bioMérieux, Marcy—l’ Étoile, France) and positive isolates were identified using MALDI-TOF Mass Spectrometry (Bruker Daltonics, Bremen, Germany) and further analyzed using the PCR technique for the presence of the following ARGs: van A, van B, tet(M), tet(L), ermB, msrA, mefA, aac(6′)-Im, aph(2)-Ib, ant(4′)-Ia, sul1, sul2, sul3, and NDM1. We isolated and identified 68 isolates of Enterococcus faecium and 11 isolates of Enterococcus faecalis. The molecular analysis showed 66 isolates positive for the vanA gene and eight positive for vanB. The most frequent association of ARG in VRE was vanA-tet(M)-ermB. There was no statistically significant difference between Enterococcus faecium and Enterococcus faecalis regarding ARGs. Our work proves that during the COVID-19 pandemic, highly resistant isolates of Enterococcus were present in patients in the intensive care unit; thus, better healthcare policies should be implemented for the management and control of these highly resistant isolates in the future.

Here, we develop a robust and sensitive real-time PCR assay which allows the simultaneous detection of vanA and vanB genes using common primers. The system was designed using the Primer3 online software. The specificity of primers and probes was first checked by in silico PCR and by BlastN analysis. The genomic DNA of 255 bacterial isolates, including Enterococcus spp., Gram-negative, and Gram-positive strains, as well as a collection of 50 stool and 50 rectal swab samples, were tested to evaluate the specificity of the new real-time PCR (RT-PCR) system. The results of the designed RT-PCR were 100% specific and 100% positive on tested vancomycin resistant isolates harboring either the vanA or vanB gene. RT-PCR assays were negative for all other bacterial species tested including vancomycin-sensitive Enterococci and Enterococcus strains harboring vanC genes. The limit of detection of vanA and vanB genes by RT-PCR assay was 47 CFU/mL and 32 CFU/mL, respectively. The rapid and accurate detection of vancomycin-resistant Enterococci is the cornerstone for minimizing the risk of nosocomial transmissions and outbreaks. We believe that this assay will strengthen routine diagnostics and surveillance programs.

Background/Purpose: Screening for vancomycin-resistant enterococci (VRE) by culture takes days to generate results, while polymerase chain reaction (PCR) testing directly from clinical specimens lacks specificity. The aims of this study were to develop a real-time PCR to detect and identify Enterococcus faecium, Enterococcus faecalis, and vanA and vanB genes, and to evaluate the impact of this PCR on test-reporting times when performing it directly from suspect VRE isolates present on screening chromogenic media. Methods: The tetraplex PCR primers were designed to amplify E. faecium, E. faecalis, and vanA and vanB genes, with melt-curve analysis of PCR products. Following analytical and clinical validation of the molecular assay, PCR testing was performed for target colonies present on VRE chromogenic media. PCR results were evaluated against conventional phenotypic identification and susceptibility testing, with the time to result being monitored for both modalities. Results: A total of 519 colonies from clinical specimens were tested concurrently by real-time PCR and phenotypic methods. In all, 223 isolates were identified with phenotypic vancomycin resistance (vanA, n = 108; vanB, n = 105; non-vanA/vanB = 10), with complete agreement between PCR and phenotypic testing for vancomycin-resistant E. faecium and E. faecalis. The majority (88.6%) of PCR results were reported, on average, 24.8 hours earlier than those of phenotypic testing, with 68% reduction in total costs. Conclusion: The use of culture on selective media, followed by direct colony PCR confirmation allows faster and economical VRE screening.

Background & Objectives: Enterococci are opportunistic bacteria that play a significant role in creating nosocomial infections. These bacteria recognized as the second cause of urinary tract infections in hospitalised and non-hospitalised patients worldwide. The aim of this study was to identify the prevalence of vancomycin resistance genes and antibiotic resistance patterns in VRE strains. Methods: This cross-sectional study was carried out on 48 Enterococcus isolates collected from urinary of hospitalised and non-hospitalised pymaniye hospital patients in south of Fars province. Enterococci separation was performed through Biochemical tests. Sensitivity to conventional antibiotics and vancomycin according to CLSI criteria was conducted by using disk diffusion method and measuring MIC through Broth Broth dilution method. Then the presence of van A, van B, van C1/C2 genes in VRE strains were measured through multiplex PCR technique. Results: Out of total isolated Enterococci, 13 (27/08%) belonged to E.faecalis, 6(12/50%) to E.faesium and 29 (60/42%) non-faecalis and non-faecium. In total, 21 strains (43/75%) were VRE and resistance to all groups of antibiotics observed in 3 (14/28%) strains. 4strains (40%) had vanA gene and 2 strains (20%) had vanB gene. But none of the strains were carrying vanC1/C2 and none of the investigated genes observed in 4 strains. Conclusion: According to wide emergence of the vancomycin resistant enterococci, application of precautionary and management procedures are highly required. Furthermore, avoidance from arbitrarily taking of antibiotics is recommended.

Introduction: Nasal colonization by coagulase-negative Staphylococci (CoNS) play an important role in nosocomial infections. This study aims to determine antibiotics susceptibility pattern and molecular screening of methicillin- and vancomycin-resistant nasal CoNS among hospitalized patients. Methodology: Nasal swabs were collected from 202 inpatients at Prince Hamzah Hospital, Jordan. Swabs were processed according to standard microbiological procedures to isolate Staphylococci. Antibiotic susceptibility testing was performed using disk diffusion, E-test, microdilution, and Vitek 2. Molecular analysis was performed using PCR for the detection of mecA, vanA, and vanB genes. Results: Nasal Staphylococci was isolated in 64/202 (31.7%) samples. Thirty isolates (14.8%) were CoNS, including S. haemolyticus (n = 17, 8.4%), S. sciuri (n = 6, 3%), S. epidermidis (n = 2, 1%), S. warneri (n = 2, 1%), S. hominis (n = 2, 1%), and S. lentus (n = 1, 0.5%). Twentytwo (10.9%) isolates were MR-CoNS harboring mecA gene. CoNS and MR-CoNS isolates were highly resistant to benzylpenicillin, erythromycin, fosfomycin, and imipenem. All isolates were sensitive to vancomycin by E-test and microdilution test and were negative for vanA and vanB genes. Nasal CoNS colonization was associated with an increased number of family members living with the participant (P = 0.04) and with admission to the orthopedic department (P = 0.03), while MR-CoNS colonization was associated with smoking (P = 0.03). Conclusions: Nasal colonization by unusual CoNS species and mecA-positive MR-CoNS are common among hospitalized patients. Absence of vanA and vanB genes suggests little contribution of nasal CoNS to vancomycin resistance transmission. [ABSTRACT FROM AUTHOR]

Background: The incidence of human infection and colonization with methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) has increased in the recent years. Environmental sources, including bird droppings, might play an important role as resistance reservoirs. Results: Fresh fecal samples were collected from rooks and wild-living birds during the autumn-winter period of 2016/2017, and tested for the presence of bacteria associated with human diseases. Besides bacteria representing the genera Enterococcus, Campylobacter, Escherichia, and Staphylococcus, Enterobacter, Citrobacter, Proteus, Hafnia, and Pseudomonas were also identified. The susceptibility of S. aureus and Enterococcus spp. isolates to methicillin, and vancomycin and teicoplanin, respectively, was analyzed to assess the avian wildlife as a reservoir of MRSA and VRE strains. Twenty-two percent of all S. aureus isolates were methicillin-resistant. These strains were screened by polymerase chain reaction (PCR), using the most widely used primer sets specific for the mecA gene. Twenty percent of all Enterococcus strains were phenotypically vancomycin-resistant. The presence of van resistance genes in these strains was investigated by PCR using vanA and vanB gene-specific primers. A good correlation between mecA gene detection and disc diffusion data was observed, while some discrepancy was noted between the PCR data and the vancomycin/teicoplanin phenotypic resistance pattern. The incidence of strains resistant to methicillin and glycopeptide antibiotics in wild-living birds was twice that in rooks. Conclusions: The study suggests that rooks from urban areas and passerine birds from the natural habitat carry antibiotic-resistant Enterococcus spp. and S. aureus strains, probably reflecting the presence of such isolates in the environmental food sources. [ABSTRACT FROM AUTHOR]

Antibiotic resistance is one of the serious threats to global public health and food safety today. Acquired antibiotic resistance in microorganisms arises from prevalent use of antibiotics for human and animal medicine. Owing to the fact that Vancomycin Resistant Enteroccocci (VRE) is a vital problem for public health, determination of the antibiotic resistance profiles of Enterococcus spp. isolates have crucial importance as a part of the farm to fork food safety. In the study, 120 retail chicken meat samples were analyzed, and 36 (30%) of the samples were detected as Enterococcus spp. positive. According to the results, the most prevalent species was E. faecalis with a rate of 44.4% (16/36), followed by 27.8% (10/36) E. faecium, 11.1% (4/36) E. durans, 2.8% (1/36) E. gallinarum and 2.8% (1/36) E. casseliflavus. Antibiotic resistance profiles of the verified Enterococcus spp. isolates were determined with disc diffusion method in terms of eight different antibiotics. Among the Enterococcus spp. isolates, 20 (55.5%) isolates were phenotypically resistant to vancomycin, 6 isolates (16.7%) were detected as vanA positive, 3 isolates (8.3%) were detected as vanB positive, and one isolate (5%) showed high resistance to vancomycin (MIC >256 µg/ml). Even though the observed percentages are low, the observed resistance patterns are still of concern for public health. [ABSTRACT FROM AUTHOR]

Purpose: According to the WHO vancomycin-resistant Enterococcus faecium (VRE) belongs to the microorganisms for which new antibiotics are urgently needed. Depending on the type of vancomycin resistance vanA gene VRE is differentiated from vanB VRE and other types. In this retrospective analysis the results of VRE surveillance performed at a German tertiary hospital with approximately 1,200 beds between 2013 and 2017 are shown.Patients and Methods: Rectal screening swabs were taken at admission and once per week on the early rehabilitation ward of Ingolstadt Hospital (ERWIN) but not at other wards. The number of VRE colonized patients was evaluated by using appropriate computer software (LabCentre, Hybase). The Hybase program was also used to find out the number of Saccharomyces boulardii and multi-susceptible Escherichia coli Nissle in blood cultures of patients at ERWIN. The mechanism of vancomycin resistance was examined by PCR and clonality of VRE strains was analyzed by pulsed-field gel electrophoresis.Results: Between 2013 and 2015 the number of VRE increased from 30 to 78 per year whereas in 2016 and 2017 the number declined to 51. Systematic analysis of the laboratory data revealed that this increase was driven by oligoclonal transmission of vanB VRE on ERWIN until August 2016 despite performing intensified infection control measures. However, afterward the number of VRE decreased at ERWIN and subsequently at the other wards. While searching for the reason behind this beneficial development we noticed that at ERWIN, patients treated with antibiotics received two probiotic medications simultaneously (S. boulardii, E. coli Nissle) for the duration of the antibiotic therapy plus an additional 2 days. There was no indication of side effects caused by these microorganisms, particularly no infections.Conclusion: Application of S. boulardii and E. coli Nissle was safe and associated with reduced transmission of VRE from patient to patient at ERWIN. Therefore, in our setting, probiotic treatment of patients receiving antibiotics contributed to the increase of patients' safety. [ABSTRACT FROM AUTHOR]

Background: The emergence of resistance among enterococci threatens to make vancomycin obsolete in the treatment of infections caused by these bacteria. Resistance is observed most commonly in Enterococcus faecium, which is often resistant to aminoglycosides and ß-lactams and to a lesser extent in other Enterococcus species, including Enterococcus faecalis. Aim: The aim of this study is to estimate the prevalence of vancomycin-resistant enterococci and study types of glycopeptide resistance genes. Materials and Methods: A total of 100 nonrepeat isolates of enterococci from various clinical samples were analyzed. As per the Clinical and Laboratory Standards Institute guidelines enterococci were screened for vancomycin resistance by Kirby-Bauer disc diffusion method. The minimum inhibitory concentration of all isolates for vancomycin was determined by Epsilometer test. Multiplex polymerase chain reaction (PCR) was carried out for vancomycin-resistance enterococcal isolates using six sets of primers to identify van genes responsible for resistance. Results: Twelve percent isolates were found to be vancomycin resistant. By multiplex PCR 100% vancomycin-resistant isolates carried vanA gene. However, vanB, vanC, vanD, vanE, vanG genes which encode other resistance ligase were not detected. Amplicons were sent for sequencing and the sequence received showed 100% identity with vanA gene. Conclusion: The prevalence of vancomycin resistance among enterococci isolates in this study was 12%. Multiplex PCR can detect van genes with high sensitivity and specificity responsible for vancomycin resistance. Treating serious infections caused by vancomycin-resistant enterococci has emerged as one of the leading clinical challenges for physicians because of limited therapeutic options.. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to determine the antimicrobial susceptibility of vancomycin-resistant and Enterococcus faecium strains isolated from urine samples of dogs. A total of 22 Enterococcus sp. samples were isolated and identified from 100 urine samples collected by cystocentesis from dogs of both sexes. The identification with species specific primers for multiplex PCR revealed that all 22 isolates (100%) belonged to E. faecium. Vancomycin resistance was found in 10 (45%) samples of E. faecium strains with PCR study by vanA and vanB primers. [ABSTRACT FROM AUTHOR]

Abstract Vancomycin-resistant enterococci (VRE) are important nosocomial pathogens. Invasive VRE infections are difficult to treat since common therapeutic options including ampicillin and glycopeptides often fail. In vitro , most VRE remain susceptible to last-resort antibiotics such as linezolid, tigecycline and daptomycin. However, neither tigecycline nor linezolid act in a bactericidal manner, and daptomycin has proven activity only at high dosages licensed for treating enterococcal endocarditis. Despite these pharmacological and therapeutic limitations, reports on resistance to these last-resort drugs in VRE, and enterococci in general, have increased in recent years. In this review, we briefly recapitulate the current knowledge on the mode of action as well as the known and novel mechanisms of resistance and describe surveillance data on resistance to linezolid, tigecycline and daptomycin in enterococci. In addition, we also suggest a common nomenclature for designating enterococci and VRE with resistances to these important last-resort antibiotics. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus (MRSA) infections in humans can currently only be treated with vancomycin. Consequently, vancomycin-resistant Enterococcus spp. pose a serious public health hazard because MRSA can acquire their vancomycin resistance. While the microbiological and genetic characteristics of vancomycin-resistant enterococci (VRE) have been extensively studied, serological diagnostic tools for these organisms are lacking. The VanA and VanB classes of VRE show marked resistance. Here, we identified the VanA and VanB proteins that are immunogenic in mice. To do so, mice were orally infected with a VanA strain of E. faecium or a VanB strain of E. faecalis and the serologically immunogenic proteins were identified by SDS-PAGE and Western blot analysis. The mice reacted to the 27 and 65 kDa cell envelope (CE) proteins of VanA at 1 week post-infection (wpi) and then reacted to the 100 kDa cytoplasmic protein (CP) at 2-4 wpi. With regard to VanB, the mice responded at 1-4 wpi, 3-4 wpi, and 4 wpi to the 70 kDa, 25 and 35 kDa, and 79 kDa CE proteins, respectively, and at 3 wpi to the 39 kDa CP. The identification of these immunogenic proteins may be useful for diagnosing and for producing immunotherapeutic VRE antibodies. [ABSTRACT FROM AUTHOR]

Introduction: Enterococci are part of the normal intestinal flora and have been recognized as important human pathogens. Vancomycin Resistant Enterococci (VRE) are global threat as this resistance is transmissible and also poses a challenge for infection control. Aim: This study was undertaken to study phenotypic and genotypic characteristics of VRE from clinically significant infections among hospitalized patients and their association with gut colonization. Materials and Methods: Clinically significant isolates of enterococci (n=250) were studied. Species confirmation was done by Polymerase Chain Reaction (PCR). Minimum Inhibitory Concentration (MIC) for vancomycin was determined by E-test. PCR for VanA, VanB and VanC1 gene was done for genotypic characterization. MIC for teicoplanin, linezolid, tigecycline, daptomycin and quinupristin-dalfopristin was determined by E test. Patients with VRE infection were screened for gut colonization using vancomycin screen agar (6 µg/mL). Continuous data was analysed using the Student’s t-test. Categorical data was assessed using Pearson’s Chi-square test. A value of p ≤ 0.05 was considered statistically significant. Results: There was good correlation between the phenotypic and genotypic methods used for species identification and detection of vancomycin resistance. E. faecium (162, 64.8%) was most common followed by E. faecalis (82, 32.84%) and E. gallinarum (6, 2.4%). Overall higher resistance was observed among E. faecium. Vancomycin MIC ≥ 2 µg/mL was noted in 63 (25.2%) isolates. Fifty seven isolates showed presence of vanA and vanC1 was detected in six isolates of E. gallinarum. Isolates with VanB genotype was not detected in the present study. MIC50 (µg/mL) for teicoplanin, linezolid, tigecycline, daptomycin and quinupristin-dalfopristrin was 24, 0.75, 0.064, 2 and 0.064 respectively. Resistance to linezolid (1, 1.6%) and tigecycline (2, 3.2%) was rare. Majority (33/47, 70.2%) patients with clinically significant VRE infection showed gut colonization. Conclusion: Vancomycin resistance among enterococci is emerging. Emergence of tigecycline and linezolid resistance is also posing a challenge for clinicians. Thus, further investigations are warranted to control vancomycin resistance among pathogens.

Objective: The purpose of this study was to determine the prevalence of vancomycin-resistant Staphylococcus aureus isolated from clinical samples in Shiraz hospitals. Methods: From March to December 2012, 100 S. aureus isolates (mainly from wound and blood) were collected from three hospitals in Shiraz, south of Iran. After identification of Staphylococcus aureus by biochemical, microbiological and molecular methods, antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion test for 13 different antibiotics. Vancomycin-resistant Staphylococcus aureus isolates were determined by vancomycin agar screening test and PCR for vancomycin resistant genes (vanA and vanB). Results: The lowest and highest resistance was seen for quinupristin-dalfopristin (n=1) and ampicillin (n=95), respectively. Vancomycin agar screening test showed that 37 isolates can grow on these media. Further study by PCR also detected vanA and/or vanB genes in all of these strains. Also, 19 isolates showed either vanA or vanB but were susceptible according to vancomycin agar screening test. In total, vanA and vanB resistant genes were detected in 34% and 37% of clinical isolates, respectively. Conclusion: The results showed that the frequency of vancomycin resistance genes (vanA, vanB) is very high in Staphylococcus aureus strains isolated from patients in south of Iran. Thus, urgent interventions are needed to keep the emergence and transmission of these isolates to a minimum.

ABSTRACT In the last decade, Clostridium difficile infection (CDI) has reached an epidemic state with increasing incidence and severity in both health care and community settings. Vancomycin is an important first-line therapy for CDI, and the emergence of resistance would have significant clinical consequences. In this study, we describe for the first time a vanB2 vancomycin resistance operon in C. difficile, isolated from an Australian veal calf at slaughter. The operon was carried on an ~42-kb element showing significant homology and synteny to Tn1549, a conjugative transposon linked with the emergence and global dissemination of vancomycin-resistant enterococci (VRE). Notably, the C. difficile strain did not show any reduced susceptibility to vancomycin in vitro (MIC, 1 mg/liter), possibly as a result of an aberrant vanRB gene. As observed for other anaerobic species of the animal gut microbiota, C. difficile may be a reservoir of clinically important vancomycin resistance genes. IMPORTANCE In an era when the development of new antimicrobial drugs is slow, vancomycin remains the preferred antimicrobial therapy for Clostridium difficile infection (CDI), the most important health care-related infection in the world today. The emergence of resistance to vancomycin would have significant consequences in relation to treating patients with CDI. In this paper, we describe for the first time a complete set of vancomycin resistance genes in C. difficile. The genes were very similar to genes found in vancomycin-resistant enterococci (VRE) that were associated with the emergence and global dissemination of this organism. Fortunately, the C. difficile strain did not show any reduced susceptibility to vancomycin in vitro (MIC, 1 mg/liter), possibly because of a small difference in one gene. However, this observation signals that we may be very close to seeing a fully vancomycin-resistant strain of C. difficile.

Vancomycin-resistant enterococci [VRE] have been isolated from municipal, hospital and agricultural wastewater, recreational beaches, wild animals, birds and food animals around the world. In this study, American crows ( Corvus brachyrhynchos ) from sewage treatment plants (WWTP), dairy farms, and a large roost in a restored wetland with corresponding environmental samples were cultured for VRE. A total of 245 samples [156 crows, 89 environmental] were collected and screened for acquired vanA , vanB and/or intrinsic vanC1 genes. Samples were enriched overnight in BHI supplemented with 20 μg/mL aztreonam, 4 μg/mL vancomycin and plated on m-Enterococcus agar media supplemented with 6 μg/mL vancomycin. Selected colonies were grown on BHI media supplemented with 18 μg/mL vancomycin. Of these, 24.5% of the crow and 55% the environmental/cow samples were VRE positive as defined by Enterococcus spp. able to grow on media supplemented with 18 μg/mL vancomycin. A total of 122 VRE isolates, 43 crow and 79 environmental isolates were screened, identified to species level using 16S sequencing and further characterized. Four vanA E. faecium and multiple vanC1 E. gallinarum were identified from crows isolated from three sites. E. faecium vanA and E. gallinarum vanC1 along with other Enterococcu s spp. carrying vanA , vanB , vanC1 were isolated from three environments. All enterococci were multidrug resistant. Crows were more likely to carry vanA E. faecium than either the cow feces or wetland waters/soils. Comparing E. gallinarum vanC1 from crows and their environment would be useful in determining whether crows share VRE strains with their environment. [ABSTRACT FROM AUTHOR]

Aim: The aim of this study was to investigate the VRE frequency and the rate of each gene in isolated enterococci from patients with intestinal infection in the central region of Iran. Background: Enterococci infections are a public health growing concern due to the glycopeptide antibiotics resistance especially vancomycin. Genes, vanA, B, and H contribute to the influence of vancomycin-resistant enterococci (VRE). Patients and methods: This study was conducted from January to July 2014 in Shahrood university hospital. Enterococci isolation and its antibacterial susceptibility were performed by culturing in Aesculin Azide agar and Kirby-Bauer method, respectively. Vancomycin-resistant genes were screened through conventional PCR, and subsequently sequenced. Results: Among 265 specimens, 100 isolates revealed enterococci, in which E. faecalis (91%) and E. faecium (9%). The isolated enterococci were resistant to vancomycin (6%) and chloramphenicol (21%), whereas their large proportions (94% to 100%) were multi-drug resistant. All VRE isolates belonged to E. faecalis, conversely, the E. faecium were susceptible to the same antibiotic. Both vanA and vanH genes were identified in all VRE isolates, although, no vanB gene was indicated. Homology analysis of sequenced amplicons verified the full length compatibility to the worldwide reported genes. Conclusion: The present study revealed VR E. faecalis in gastroenteritis patients and resistance factor for vanA and vanH genes are coordinated. Since enterococci isolates were all multidrug resistance, increase in VR E. faecalis vanA / vanH in this area could be expected. [ABSTRACT FROM AUTHOR]

The emergence of clinical enterococcal isolates that are resistant to both ampicillin and vancomycin is a cause of great concern, as therapeutic alternatives for the treatment of infections caused by such organisms are becoming limited. Aquatic environments could play a role in the dissemination of antibiotic resistant enterococci. This study investigated the presence of ampicillin and vancomycin resistant enterococci in the treated effluent of six wastewater treatment plants (WWTPs) and in surface water used as a source for drinking water production in the Netherlands. Membrane filtration in combination with selective media with ampicillin or vancomycin was applied to determine the presence of ampicillin resistant Enterococcus (ARE) and vancomycin resistant Enterococcus (VRE) species. Ampicillin resistant Enterococcus faecium (minimal inhibitory concentration (MIC) >16μg/mL; n=1033) was observed in all studied WWTP effluents. In surface water used for drinking water production (intake locations), no ARE or VRE were observed. At both types of location, intrinsic vancomycin resistant Pediococcus spp., Leuconostoc spp. and Lactobacillus spp. were isolated with the vancomycin medium. The ampicillin resistant E. faecium (AREfm) isolates (n=113) did not contain the vanA or vanB gene, but MIC testing for vancomycin showed intermediate vancomycin resistance (2-8μgmL(-1)) to occur in these AREfm strains. This study documents the discharge of ampicillin resistant E. faecium strains with intermediate vancomycin resistance by the WWTPs into the surface water, but no presence of these strains downstream at intake locations for drinking water production. [ABSTRACT FROM AUTHOR]