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Mikiyas Zike,1 Abdurahaman Mohammed Ahmed,2 Awraris Hailu,2 Bedru Hussien3 1Debre Berhan Town Health Facilities, MoH, Debre Berhan, Amhara National Regional State, Ethiopia; 2Department of Epidemiology, Debre Berhan University, Debre Berhan, Amhara National Regional State, Ethiopia; 3Department of Public Health, Madda Walabu University, Goba, Oromia National Regional State, EthiopiaCorrespondence: Bedru Hussien, Department of Public Health, Madda Walabu University, P.O. Box 302, Goba, Oromia National Regional State, Ethiopia, Tel +251911997287, Email bedru1964@gmail.comBackground: The majority of multidrug-resistant organisms found in immunocompromised patients are enterococci. The rise of vancomycin-resistant enterococci (VRE) poses a significant threat to public health. There is a scarcity of information regarding the prevalence of VRE in Ethiopia.Purpose: This study aims to determine the prevalence of VRE in fecal samples from Human Immunodeficiency Virus (HIV)-positive individuals, to identify associated factors, and to assess their susceptibility to selected commonly prescribed medications.Patients and Methods: A cross-sectional study was conducted from April 1 to July 15, 2023, on 170 HIV-positive clients at Debre Berhan Town. A pre-tested structured questionnaire was used to collect socio-demographic and clinical data. Stool sample was collected by trained health workers, and processed by standard microbiological techniques. Kirby-Bauer disk diffusion method was used for antimicrobial susceptibility testing. Data entry and analysis was carried out by SPSS Version 25. Bivariate and multivariate logistic regressions were used to assess the associated factors. Variables with a p-value of < 0.05 were considered to be significantly associated with the outcome variables and the results were displayed with tables.Results: From the total of 170 study participants, colonization of Enterococcus species was observed among 95 (55.9%). Vancomycin resistance was found in 13 (13.8%) of them with 95% confidence interval (CI) 7.4– 22.1. History of hospitalization Adjusted Odds Ratio (AOR): 11.9 (95% CI 1.11– 127.53); habit of eating uncooked food (AOR: 15.34 (95% CI 2.36– 99.63)) and invasive procedures (AOR: 23.07 (95% CI 3.54– 150)) were among the predictors of VRE. MDR (multidrug resistance) was observed in 83 (87.4%) of the isolates. The highest rate of resistance was observed for ampicillin with 72 (74.6%).Conclusion: Vancomycin and multidrug resistance of enterococci among HIV patients are significant in ART clinics of Debre Berhan Town. These warrant applicable infection prevention guidelines in the health facilities and health education on food hygiene.Keywords: Enterococci, vancomycin resistant, HIV patients, Ethiopia

DEADLY pathogenic multidrug-resistant bacteria (MDR) are becoming more prevalent every day and represent a major danger to human health. This research aimed to isolate and quantify vancomycin resistant MDR (VRMDR) Enterococcus faecalis and extended-spectrum β-lactamase MDR (ESBLMDR) Klebsiella pneumoniae by detecting specific genes and the use of TiO2 Aspergillus oryzae nanoparticles as antibacterial agent against the two strains. One hundred and fifty clinical specimens were collected from Mbarret El-Asafra Hospital, 80 isolates were E. faecalis and 70 isolates were K. pneumoniae. 21/80 was found to be VRMDR E. faecalis, the findings showed that 76.19% of VRMDR E. faecalis strains harboured the VanA gene and 90.47% harboured the VanB gene, while 66.66% of them carried the two resistance genes. On the other hand, 18/70 samples were found to be ESBLMDR K. pneumoniae, the findings showed that 72.22% of ESBLMDR K. pneumoniae strains harboured the blaTEM gene and 61.11% harboured the blaSHV gene, while 33.33% of them carried the two resistance genes. 30µg/ml of nano TiO2 A. oryzae was found to be the minimal inhibitory concentration (MIC) for the VRMDR E. faecalis, while 50µg/ml of nano TiO2 A. oryzae was found to be the MIC for the ESBLMDR K. pneumoniae. The IC50 of TiO2 A. oryzae nanoparticles against human gastric epithelial cell line (GES1) was 563.023±31.7µg/ml compared to chloramphenicol, imipenem drugs and TiO2 nanoparticles showing (563.023±31.7µg/ml, 169.386±9.32µg/ml, 71.692±5.05µg/ml and 30.562±3.22µg/ml) respectively, showing that TiO2 nanoparticles, chloramphenicol and imipenem were more cytotoxic on GES1 normal cells than TiO2 A. oryzae nanoparticles. [ABSTRACT FROM AUTHOR]

Zhirou Xiao,1,&ast; Liping Qu,2,&ast; Haijun Chen,1 Wanting Liu,2 Yi Zhan,2 Jiahui Ling,2 Hongwei Shen,3 Ling Yang,1 Dingqiang Chen2 1Department of Laboratory Medicine, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, People’s Republic of China; 2Microbiome Medicine Center, Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China; 3Department of Clinical Laboratory, Shenzhen Hospital of Southern Medical University, Guangzhou, Guangdong, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ling Yang, Department of Laboratory Medicine, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510120, People’s Republic of China, Tel/Fax +86-20-83062158, Email jykresearch@126.com Dingqiang Chen, Microbiome Medicine Center, Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, 510282, People’s Republic of China, Tel/Fax +86-20-62783335, Email jyksys@126.comBackground: Antibiotic resistance represents a serious global health challenge, particularly with the emergence of strains resistant to last-resort antibiotics such as tigecycline, polymyxin B, and vancomycin. Urgent measures are required to alleviate this situation. To facilitate the judicious use of antibiotics, rapid and precise antimicrobial susceptibility testing (AST) is essential. Heavy water (deuterium oxide, D2O)-labeled Raman spectroscopy has emerged as a promising time-saving tool for microbiological testing.Methods: Deuterium incorporation and experimental conditions were examined to develop and apply a Raman-based AST method to evaluate the efficacy of last-resort antibiotics, including tigecycline, polymyxin B, and vancomycin, against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus faecium. Essential agreement and categorical agreement were used to assess the metabolism inactivation concentration based on Raman spectroscopy (R-MIC)–a new metric developed in this study—and minimum inhibitory concentration (MIC) determined via the traditional microdilution broth method. Spearman’s rank correlation coefficient was employed to measure the association between R-MIC and MIC values.Results: The Raman-based AST method achieved a 100% categorical agreement (92/92) with the traditional microdilution broth method within five hours, while the traditional method required approximately 24 h. The R-MIC values shared 68.5% (63/92) consistency with the MIC values. In addition, the R-MIC and MIC values were highly correlated (Spearman’s r=0.96), resulting in an essential agreement of 100%.Conclusion: Our optimized experimental method and conditions indicate that Raman-based AST holds great promise as a solution to overcome the time-consuming challenges of traditional AST methods.Keywords: Raman spectroscopy, tigecycline resistant, polymyxin B resistant, vancomycin resistant, antimicrobial susceptibility testing

Aims: Enterococcus bacteria, including some strains that are resistant to antibiotics like vancomycin can pose a threat to public health. The purpose of this study is to identify the species, antibiotic susceptibility profile and VanA/VanB gene frequencies in Enterococci isolated from patients at a tertiary hospital in Kota Kinabalu, Sabah. Methodology and results: Various bodily fluid specimens were collected from 162 patients between July 2019 and June 2021. Species confirmation and susceptibility testing were performed using an automated system. Subsequently, PCR was used to determine the presence of VanA and VanB genes. Species identification revealed the presence of five enterococcal species, namely E. faecalis (91), E. faecium (64), E. gallinarum (3) E. casseliflavus (2), along with one isolate each of E. hirae and E. avium. Overall, resistance to antibiotics like ampicillin, quinolones, tetracycline, gentamicin-syn, nitrofurantoin, glycopeptides and linezolid was generally low (<50%). However, a significant number of isolates displayed high resistance to erythromycin (>50% of samples), while resistance to tetracycline was more moderate. The frequencies of VanA and VanB genes were low (0.6 and 0%, respectively) and they were only detected in E. faecium. Conclusion, significance and impact of study: The results indicate that while the prevalence of vancomycin-resistant enterococci (VRE) may be low, there is an increasing incidence of multidrug-resistant enterococci, particularly with regards to erythromycin. [ABSTRACT FROM AUTHOR]

Introduction: The burden of community acquired antibiotic resistance is increasing with an alarming rate in the developing countries. Many factors, including inappropriate use of antibiotics, have been suggested as the causes of spread of resistant strains in the community. Aim: To compare antibiotics susceptibility pattern of community acquired gram positive isolates among women of reproductive age with history of recent inappropriate antibiotics use and those with no history of antibiotics use. Materials and Methods: The present study was a cross-sectional descriptive study carried out from August 2020 to February 2021 at the State Teaching Hospital in Enugu, Nigeria. The participants for the study were 713 apparently healthy women who presented to the family planning unit for preconception care. All the participants were screened for asymptomatic bacteriuria by culturing their Midstream Urine (MSU). The isolated organisms were identified, and Antimicrobial Susceptibility (AST) test performed using the Vitek 2. Polymerase Chain Reaction (PCR) was done for the presence of mecA gene among methicillin resistant Staphylococcus species. The antibiotic susceptibility pattern of the isolates from participants with positive history of recent antibiotic use was compared with the susceptibility pattern of those with no history of antibiotics using Chi-square test. Results: Out of the 713 participants, 59 (8.3%) had a positive history of recent antibiotics use. Only 1 (1.7%) participant had her antibiotics prescribed by the doctor while 13 (22) and 45 (76.3) obtain their antibiotics at the patent medicine dealer shop and through self-medication respectively. The commonest indication for inappropriate use was febrile illness followed by upper respiratory symptoms. Ciprofloxacin was the most abused antibiotics followed by amoxicillin-clavulanic acid. Twenty (83.3%) and 4 (16.7%) had Asymptomatic Bacteriuria (ASB) among those with recent antibiotic use and those with no history of recent antibiotic use respectively. All the Methicillin Resistant Staphylococcus aureus (MRSA) was isolated from the participants with recent history of inappropriate antibiotic use. Also, of significant among this group were quinolone resistant gram positive organisms. Conclusion: ASB and multidrug resistant gram positive isolates were common among the participants with recent history of inappropriate antibiotic use. There is need to regulate antibiotics use in the community to prevent selection of multidrug resistant organisms. [ABSTRACT FROM AUTHOR]

Enterococci, the normal inhabitant of gastrointestinal tract of humans and animals, have emerged as significant antibiotic resistant nosocomial pathogens. The current study was designed to determine the antibiotic resistance profile and genes harbored by isolated strains of Enterococci along with study of antibiotic resistance transfer potential from resistant Enterococci to susceptible pathogenic Staphylococcus aureus in vitro. The PCR based prevalence of Enterococcus faecalis and Enterococcus faecium from 118 broiler cloacal swabs was 60.46 and 30.23%, respectively, indicating that E. faecalis is the predominant species in broilers followed by E. faecium. Enterococci (n=86) were examined for the phenotypic resistance against eleven antibiotics which showed higher level of resistance to lincomycin (96.51%), erythromycin (90.69%), tetracycline (86.04%) and streptomycin (75.58%), intermediate level of resistance to ciprofloxacin (54.65%) and doxycycline (48.83%), and low resistance level to penicillin (26.74%), chloramphenicol (26.74%), amoxicillin (17.44%), augmentin (11.62%) and vancomycin (10.46%). Over 80% Enterococcal isolates were found multidrug resistant (MDR). On the basis of PCR analysis, erm (B) and tet (M) genes were identified in all phenotypically erythromycin and tetracycline resistant strains while van (B) was identified in only 4/9 (44.4%) of vancomycin resistant enterococci (VRE) with no detection of van (A) gene in any VRE. One strain E. faecalis (FME-41) was able to transfer the erythromycin resistance to pathogenic Staphylococcus aureus (M-11) in broth mating assay. MDR Enterococci pose therapeutic threat to human community and control on the spread of such MDR Enterococci from poultry to human food chain is crucial. [ABSTRACT FROM AUTHOR]

Introduction: The burden of community acquired antibiotic resistance is increasing with an alarming rate in the developing countries. Many factors, including inappropriate use of antibiotics, have been suggested as the causes of spread of resistant strains in the community. Aim: To compare antibiotics susceptibility pattern of community acquired gram positive isolates among women of reproductive age with history of recent inappropriate antibiotics use and those with no history of antibiotics use. Materials and Methods: The present study was a cross-sectional descriptive study carried out from August 2020 to February 2021 at the State Teaching Hospital in Enugu, Nigeria. The participants for the study were 713 apparently healthy women who presented to the family planning unit for preconception care. All the participants were screened for asymptomatic bacteriuria by culturing their Midstream Urine (MSU). The isolated organisms were identified and Antimicrobial Susceptibility (AST) test performed using the Vitek 2. Polymerase Chain Reaction (PCR) was done for the presence of mecA gene among methicillin resistant Staphylococcus species. The antibiotic susceptibility pattern of the isolates from participants with positive history of recent antibiotic use was compared with the susceptibility pattern of those with no history of antibiotics using Chi-square test. Results: Out of the 713 participants, 59 (8.3%) had a positive history of recent antibiotics use. Only 1 (1.7%) participant had her antibiotics prescribed by the doctor while 13 (22%) and 45 (76.3%) obtain their antibiotics at the patent medicine dealer shop and through self-medication respectively. The commonest indication for inappropriate use was febrile illness followed by upper respiratory symptoms. Ciprofloxacin was the most abused antibiotics followed by amoxicillin-clavulanic acid. A 20 (83.3%) and 4 (16.7%) had Asymptomatic Bacteriuria (ASB) among those with recent antibiotic use and those with no history of recent antibiotic use respectively. All the Methicillin Resistant Staphylococcus aureus (MRSA) was isolated from the participants with recent history of inappropriate antibiotic use. Also, of significant among this group were quinolone resistant gram positive organisms. Conclusion: The ASB and multidrug resistant gram positive isolates were common among the participants with recent history of inappropriate antibiotic use. There is need to regulate antibiotics use in the community to prevent selection of multidrug resistant organisms.

Background & Aims: The increasing rate of vancomycin resistant Staphylococcus aureus (VRSA) with biofilm formation may become a new threat to humans. In such cases, finding an effective treatment strategy such as using Nanotechnology (Nano- drugs) to deal with these types of infections may be promising. This study aimed to investigate the inhibitory effects of silver nanoparticles (SNPs) on biofilm formation of VRSAs. Materials and Methods: Clinical S. aureus isolates were identified to the species level by conventional methods, and their identities were later confirmed by PCR. Following the determination of susceptibility patterns of the isolates; all the screened S.aureus isolates have been assessed regarding their susceptibility to vancomycin. Detection of vanA gene and determination of minimum inhibitory concentrations (MICs) of VRSAs were carried out using PCR and Etest methods, respectively. The biofilm production was assessed on all VRSA isolates in the presence/absence of SNPs using micro-titer plate method. Results: In total, 11 (6.21%) VRSAs were identified among 177 S. aureus clinical isolates. These isolates were included in the biofilm production assay. All of the VRSAs were multidrug resistance and biofilm producers. The inhibitory effect of SNPs in concentration of 250 µg/ml on biofilm formation of VRSA isolates was significant (Pv = 0.01). Conclusion: Based on our findings, SNPs can prevent biofilm formation of VRSAs and applying of these nanoparticles may prohibit from the persistence and colonization of such resistant isolates.

Abstract Background Even though enterococci can cause serious infections in multiple sites, they are a rare cause of pneumonia. We reported a uremic patient with vancomycin-resistant E. faecium (VRE-fm) pneumonia, possibly related to epileptic seizures. Case presentation A 57-year old man with uremia on hemodialysis was admitted to the hospital with complaint of recurrent epileptic seizures, followed by a two-week history of recurrent fever and cough with purulent sputum. Chest CT demonstrated multiple exudation of both lungs. He was diagnosed as community acquired pneumonia. Despite antibiotic combination therapy, abnormal chest shadows aggravated. Sputum and blood cultures were initially negative, but later blood culture grew VRE-fm. We suspected aspiration of gastrointestinal content induced by epilepsy as the most likely mechanism. The patient was successfully treated with a four-week course of linezolid according to the antibiotic susceptibility testing. Conclusions Physicians should consider multi-drug resistant organisms such as VRE in uremic patients with pneumonia that fails to resolve with broad-spectrum antibiotics, especially in the cases with aspiration induced by epilepsy, immunocompromised conditions, and repeated or prolonged hospitalizations.

Enterococcus faecium , a gram-positive opportunistic pathogen, has become a major concern for nosocomial infections due to its resistance to several antibiotics, including vancomycin. Finding novel alternatives for treatment prevention, such as vaccines, is therefore crucial. In this study, we used various techniques to discover a novel capsular polysaccharide. Firstly, we identified an encapsulated E. faecium strain by evaluating the opsonophagocytic activity of fifteen strains with antibodies targeting the well-known lipoteichoic acid antigen. This activity was attributed to an unknown polysaccharide. We then prepared a crude cell wall glycopolymer and fractionated it, guided by immunodot-blot analysis. The most immunoreactive fractions were used for opsonophagocytic inhibition assays. The fraction containing the inhibitory polysaccharide underwent structural characterization using NMR and chemical analyses. The elucidated structure presents a branched repeating unit, with the linear part being: →)-β- d -Gal-(1 → 4)-β- d -Glc-(1 → 4)-β- d -Gal-(1 → 4)-β- d -GlcNAc-(1→, further decorated with a terminal α- d -Glc and a d -phosphoglycerol moiety, attached to O-2 and O-3 of the 4-linked Gal unit, respectively. This polysaccharide was conjugated to BSA and the synthetic glycoprotein used to immunize mice. The resulting sera exhibited good opsonic activity, suggesting its potential as a vaccine antigen. In conclusion, our effector-function-based approach successfully identified an immunogenic capsular polysaccharide with promising applications in immunotherapy. [Display omitted] [ABSTRACT FROM AUTHOR]

Mehrdad Zalipour,1 Bahram Nasr Esfahani,1 Mehrdad Halaji,1 Amir Azimian,2 Seyed Asghar Havaei1,3 1Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran; 2Department of Pathobiology and Laboratory Sciences, North Khorasan University of Medical Sciences, Bojnurd, Iran; 3Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, IranCorrespondence: Seyed Asghar HavaeiDepartment of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Hezar-Jerib Avenue, Isfahan 81746 73461, IranTel +983137922478Email havaei@med.mui.ac.irPurpose: Over the past two decades, enterococci have emerged as an important opportunistic pathogen causing life-threatening infections in hospitals. The purpose of the present study was to examine the prevalence of genes encoding virulence factor and molecular characterization of vancomycin-resistant E. faecalis strains isolated from hospitalized patients in Isfahan, the central city of Iran.Patients and methods: A total of 53 vancomycin-resistant E. faecalis isolates (VRE) obtained from clinical samples of hospitalized patients were characterized by phenotypic and genotypic methods, and 25 selected VRE isolates from internal and ICU wards were typed by multilocus sequence typing.Results: The efa was the most prevalent virulence gene (100%) among isolates, followed by gelE (92.45%), asa1 (90.56%), ace (86.79%), esp (75.47%), cylA (39.62%), and hyl (18.86%). More than 80% of the isolates were HLGR. Multilocus sequence typing showed eight different sequence types including ST6, ST422, ST28, ST448, ST531, ST328, ST421, and ST495. STs were grouped into two clonal complex (CC) including CCA (ST6, ST422, ST448, ST531) and CCF (ST28, ST421) and two singletons (ST328, ST495).Conclusion: Our data indicated a high prevalence of virulence genes among STs described in this study. In addition, the molecular analysis demonstrated a relatively high genetic diversity among selected VRE strains from the ICU in comparison with the internal ward. Therefore, in order to prevent the colonization of virulent strains in the hospital environment, infection control procedures should be performed.Keywords: Enterococcus faecalis, virulence factors, vancomycin resistant, multilocus sequence typing, Iran

Abstract Objective The objective of this study was to determine the frequency of the antimicrobial resistance and genes encoding virulence factors of enterococci isolated in hospitalized burn patients in a major burn center in Ahvaz, southwest of Iran. A total of 340 bacterial isolates were collected from the burn center from February 2014 to February 2015. The antimicrobial susceptibility and MIC of vancomycin were determined using the disk diffusion and micro-agar dilution techniques. The genus and species-specific genes, potential virulence genes, and vanA and vanB genes were detected by polymerase chain reaction. Results According to our results, out of the 340 bacterial isolates, 16.4% (n = 56) were identified as enterococci. Out of the 56 enterococcal isolates, 35 (62.5%) were Enterococcus faecalis and 21 (37.5%) were Enterococcus faecium. More than 20% (n = 5) of E. faecium demonstrated resistance to vancomycin. The gelE and asa genes were the most prevalent virulence genes in E. faecalis (48.5%) and E. faecium (43%) isolates. The emergence of vancomycin resistant E. faecium strains which have several virulence factors should be considered as a major cause of concern for burn centers. Control and management of infections induced by enterococci should be regarded as highly important in burn patients.

Background: Even though enterococci can cause serious infections in multiple sites, they are a rare cause of pneumonia. We reported a uremic patient with vancomycin-resistant E. faecium (VRE-fm) pneumonia, possibly related to epileptic seizures.Case Presentation: A 57-year old man with uremia on hemodialysis was admitted to the hospital with complaint of recurrent epileptic seizures, followed by a two-week history of recurrent fever and cough with purulent sputum. Chest CT demonstrated multiple exudation of both lungs. He was diagnosed as community acquired pneumonia. Despite antibiotic combination therapy, abnormal chest shadows aggravated. Sputum and blood cultures were initially negative, but later blood culture grew VRE-fm. We suspected aspiration of gastrointestinal content induced by epilepsy as the most likely mechanism. The patient was successfully treated with a four-week course of linezolid according to the antibiotic susceptibility testing.Conclusions: Physicians should consider multi-drug resistant organisms such as VRE in uremic patients with pneumonia that fails to resolve with broad-spectrum antibiotics, especially in the cases with aspiration induced by epilepsy, immunocompromised conditions, and repeated or prolonged hospitalizations. [ABSTRACT FROM AUTHOR]

Vancomycin-resistant enterococci (VRE) are Gram-positive bacteria that are resistant to vancomycin. Infections caused by VRE are associated with adverse outcomes such as extended length of hospital stay, increased cost, and high mortality rate. Clinical and surveillance VRE isolates have been reported from Saudi Arabia. In this review, prevalence, characteristics, risk factors, resistance, and virulence of VRE isolates associated with infection and colonization in Saudi Arabia are described. [ABSTRACT FROM AUTHOR]

Background: Tigecycline is a new, semisynthetic glycylcycline. It is active against important multidrug resistant pathogens. Aim: The purpose of this study was to investigate the sensitivity of multidrug-resistant enterococci to tigecycline, and to test the correlation between the minimal inhibitory concentration (MIC) and disk diffusion methods. Materials and methods: The antimicrobial sensitivity of 108 multidrug-resistant isolates, which included 52 vancomycin-resistant enterococci (VRE) and 56 high-level aminoglycoside-resistant (HLAR) enterococci, was tested by the E test, broth microdilution test and disk diffusion methods. Results: All of the isolates were sensitive to tigecycline, as determined by the E test and broth microdilution test. The MIC 90 value (0.19 μg/mL) of tigecycline for HLAR enterococci was higher than that for VRE (0.094 μg/mL). When results were evaluated according to species, the MIC values of tigecycline for Enterococcus faecalis were higher than those for the other species. Eleven (10.1%) isolates produced false resistance results (zone diameter ⩽15 mm) by the disk diffusion method. These cases were classified as major errors. Eight (7.4%) isolates had intermediate sensitivity (sensitivity zone of 16 or 17 mm), which were classified as minor errors. The major and minor error percentages of HLAR enterococci (14.2% major, 10.7% minor error) were higher than those of VRE (5.7% major, 3.8% minor error). These results indicate that tigecycline is effective against multidrug-resistant enterococci. The sensitivity of multidrug-resistant enterococci to tigecycline should be investigated by MIC methods. The disk diffusion method causes major errors, especially for HLAR enterococci.

The study was aimed to characterize enterococci from various clinical specimens, to determine the antimicrobial susceptibility pattern, and to explore the association between virulence factors and antimicrobial resistance. A total of 283 clinical enterococcal isolates were speciated and subjected to antimicrobial susceptibility testing. Virulence factors (hemolysin, gelatinase, and biofilm production) were detected phenotypically. Of the 283 enterococci isolated, 12 species were identified; predominant species were Enterococcus faecalis (82.33%). High-level gentamicin (HLG) and vancomycin resistance were observed among 55.57% and 6.01% of enteroccal isolates, respectively. All vancomycin-resistant enterococci (VREs) were E. faecalis and had VanA phenotype and genotype. Hemolysin, gelatinase, and biofilm production were seen in 15.90%, 12.36%, and 13.43% of enterococcal isolates, respectively. Vancomycin and HLG resistance were observed in 0.35% and 61.86% of the enterococcal isolates producing virulence factors. Isolates resistant to HLG but susceptible to vancomycin expressed more virulent factors. Further research is required to reveal the complex interplay between drug resistance and virulence factors.

ABSTRACT Vancomycin-resistant Enterococcus faecium (VREfm) is a major cause of nosocomial infection and is categorized as high priority by the World Health Organization global priority list of antibiotic-resistant bacteria. In the past, livestock have been proposed as a putative reservoir for drug-resistant E. faecium strains that infect humans, and isolates of the same lineage have been found in both reservoirs. We undertook cross-sectional surveys to isolate E. faecium (including VREfm) from livestock farms, retail meat, and wastewater treatment plants in the United Kingdom. More than 600 isolates from these sources were sequenced, and their relatedness and antibiotic resistance genes were compared with genomes of almost 800 E. faecium isolates from patients with bloodstream infection in the United Kingdom and Ireland. E. faecium was isolated from 28/29 farms; none of these isolates were VREfm, suggesting a decrease in VREfm prevalence since the last UK livestock survey in 2003. However, VREfm was isolated from 1% to 2% of retail meat products and was ubiquitous in wastewater treatment plants. Phylogenetic comparison demonstrated that the majority of human and livestock-related isolates were genetically distinct, although pig isolates from three farms were more genetically related to human isolates from 2001 to 2004 (minimum of 50 single-nucleotide polymorphisms [SNPs]). Analysis of accessory (variable) genes added further evidence for distinct niche adaptation. An analysis of acquired antibiotic resistance genes and their variants revealed limited sharing between humans and livestock. Our findings indicate that the majority of E. faecium strains infecting patients are largely distinct from those from livestock in this setting, with limited sharing of strains and resistance genes. IMPORTANCE The rise in rates of human infection caused by vancomycin-resistant Enterococcus faecium (VREfm) strains between 1988 to the 2000s in Europe was suggested to be associated with acquisition from livestock. As a result, the European Union banned the use of the glycopeptide drug avoparcin as a growth promoter in livestock feed. While some studies reported a decrease in VREfm in livestock, others reported no reduction. Here, we report the first livestock VREfm prevalence survey in the UK since 2003 and the first large-scale study using whole-genome sequencing to investigate the relationship between E. faecium strains in livestock and humans. We found a low prevalence of VREfm in retail meat and limited evidence for recent sharing of strains between livestock and humans with bloodstream infection. There was evidence for limited sharing of genes encoding antibiotic resistance between these reservoirs, a finding which requires further research.

A 66-year-old male patient who is a known case of chronic kidney disease was admitted to the hospital with complaint of seizures. He was uremic and showed signs of encephalopathy. He was previously hospitalized frequently and was on antitubercular treatment. A left sided radio cephalic fistula was made and the patient was on dialysis. Urinary catheter and central venous catheter were placed. Urine and blood cultures showed growth of Enterococcus faecium which was resistant to vancomycin – Vancomycin Resistant Enterococci (VRE) or Glycopeptide Resistant Enterococci (GRE) by antibiotic susceptibility testing. The isolate was genotyped for detection of Van A, Van B and Van C genes and was found to harbour Van A gene. The treatment details of the patient and his outcome have been described. As Enterococci are inherently resistant to many drugs, their treatment is quite challenging. Combination therapy and de-escalation from broad spectrum antimicrobials to specific narrow spectrum antibiotics immediately after antibiotic sensitivity report is available which prevents emergence of drug resistance. Spread of VRE infection must be controlled by strict infection control methods as plasmid mediated rapid transfer of VRE can occur to other organisms like Staphylococci which is a serious threat.

Introduction and Aims Rosemary (Rosmarinus officinalis) belongs to Lumiaceae family and contains different properties and compounds. With emergence of Vancomycin resistance Staphylococcus aureus, it has been confirmed as a global issue. The aim of this study is the survey of methanolic extract of rosemary leaves on Vancomycin -resistant Staphylococcus aureus isolated from patients of Imam Reza hospital of Mashhad. Materials and Methods 150 bacterial samples have collected from Imam Reza hospital of Mashhad at the beginning of 2013 November. One of them was being resistance to Vancomycin antibiotic by Disk Diffusion and MIC (Minimum Inhibitory Concentration). Extracting from Rosemary was performed by Maceration method. The MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal concentration) has evaluated by reference method of Microdilution Broth. Results The result of this study demonstrated that the MIC and MBC for Methanolic extract of Rosemary on Vancomycin -resistant Staphylococcus aureus were 0.156×103 and 0.312×103 mg/ml respectively. Conclusion Bacterial resisting to antibiotics implies necessity of using of new compounds with antibacterial property. The methanolic extract of rosemary can inhibit the growth of Vancomycin resistance Staphylococcus aureus. Therefore it can be used as clinical purposes. * Corresponding Author: Academic Center for Education, Culture and Research (ACECR) Khorasan Razavi, Center of Pathological and Medical Diagnostic Services of Neyshabur. E-mail: Mohammadsalehi73@gmail.com

Objectives: Nosocomial infections caused by methicillin resistant Staphylococcus aureus (MRSA) and emergence of vancomycin resistant S. aureus (VRSA) have led to great concern in healthcare settings worldwide. Methods: A total of 100 S. aureus clinical isolates from hospitalized patients were investigated for antimicrobial susceptibility, the presence of resistance (mecA, vanA, and vanB) and virulence (hlaA, fnbpA, cna, clfA, tsst-1, eta, and spa) encoding genes, and molecular typing based on polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) of spa gene. Results: All isolates were resistant to one or more antibiotics, with the most frequent resistance found against amoxicillin (69%). A total of 46 isolates were MRSA, and 40% of isolates were multidrug resistant (MDR). Of all S. aureus isolates, two isolates were confirmed as VRSA and four isolates confirmed as vancomycin intermediate S. aureus (VISA). The frequency of clfA, cna, tsst-1, and eta genes among MRSA isolates was significantly higher than methicillin sensitive S. aureus (MSSA). The significant correlation between MDR isolates and the carriage of multiple virulence genes was seen. All MDR isolates carried at least four virulence genes. Furthermore, biofilm formation in MRSA isolates was significantly higher than MSSA. The spa gene PCR products generated 4 major and 10 minor types. After digestion of spa amplicons with HindIII restriction enzyme, 10 different patterns ranging 174–938 bp were detected. S2b and S2a subtypes were detected frequently in MRSA isolates. Conclusions: It seems that the appropriate surveillance and control measures are essential to prevent the emergence and transmission of MRSA and VRSA strains in our country. [ABSTRACT FROM AUTHOR]

Objective: To determine the frequency of vancomycin resistant Staphylococcus aureus (VRSA) among clinical isolates of methicillin resistant Staphylococcus aureus (MRSA). Study Design: Descriptive study. Place and Duration of Study: Department of Microbiology, University of Health Sciences, Lahore from Jul 2014 to Dec 2014. Material and Methods: A total of 240 (n=240) clinical isolates of MRSA were collected by consecutive sampling from different tertiary care hospitals of Lahore. Re-confirmation of MRSA was done by the standard microbiological methods using disc diffusion technique according to Clinical Laboratory Standards Institute (CLSI) guidelines 2014. Minimum inhibitory concentration (MIC) of the vancomycin was done by agar dilution method. Results: It was found that vancomycin inhibited MRSA strains in the range of 1.0-2.0 μg/ml. Ninety percent (90%) of the strains inhibited at 1 μg/ml while 25 (10.41%) strains showed growth at 1 μg/ml which indicates that their MIC was 2 μg/ml. No vancomycin resistant (VRSA) or intermediate strains (VISA) of MRSA were found during the study but there were significant numbers of isolates having ≥1 μg/ml MIC of vancomycin. Conclusion: Vancomycin has until now excellent activity against clinical isolates of Methicillin-resistant Staphylococcus aureus. [ABSTRACT FROM AUTHOR]

Objective: The objective of this study was to determine the frequency of the antimicrobial resistance and genes encoding virulence factors of enterococci isolated in hospitalized burn patients in a major burn center in Ahvaz, southwest of Iran. A total of 340 bacterial isolates were collected from the burn center from February 2014 to February 2015. The antimicrobial susceptibility and MIC of vancomycin were determined using the disk diffusion and microagar dilution techniques. The genus and species-specific genes, potential virulence genes, and vanA and vanB genes were detected by polymerase chain reaction. Results: According to our results, out of the 340 bacterial isolates, 16.4% (n = 56) were identified as enterococci. Out of the 56 enterococcal isolates, 35 (62.5%) were Enterococcus faecalis and 21 (37.5%) were Enterococcus faecium. More than 20% (n = 5) of E. faecium demonstrated resistance to vancomycin. The gelE and asa genes were the most prevalent virulence genes in E. faecalis (48.5%) and E. faecium (43%) isolates. The emergence of vancomycin resistant E. faecium strains which have several virulence factors should be considered as a major cause of concern for burn centers. Control and management of infections induced by enterococci should be regarded as highly important in burn patients. [ABSTRACT FROM AUTHOR]

From early 2012, a novel clone of vancomycin resistant Enterococcus faecium (assigned the multi locus sequence type ST796) was simultaneously isolated from geographically separate hospitals in south eastern Australia and New Zealand. Here we describe the complete genome sequence of Ef_aus0233, a representative ST796 E. faecium isolate. We used PacBio single molecule real-time sequencing to establish a high quality, fully assembled genome comprising a circular chromosome of 2,888,087 bp and five plasmids. Comparison of Ef_aus0233 to other E. faecium genomes shows Ef_aus0233 is a member of the epidemic hospital-adapted lineage and has evolved from an ST555-like ancestral progenitor by the accumulation or modification of five mosaic plasmids and five putative prophage, acquisition of two cryptic genomic islands, accrued chromosomal single nucleotide polymorphisms and a 80 kb region of recombination, also gaining Tn1549 and Tn916, transposons conferring resistance to vancomycin and tetracycline respectively. The genomic dissection of this new clone presented here underscores the propensity of the hospital E. faecium lineage to change, presumably in response to the specific conditions of hospital and healthcare environments.

Oral streptococci are the major group of microbes isolated from oral microflora. They represent frequent pathogens of infective endocarditis (IE), and it is assumed that in most of the cases oral streptococci are acquired via mucosa layer of oral cavity. Staphylococcus aureus is also frequently isolated from IE as it accounts for 20%–30% of all cases. Vancomycin has been the most reliable therapeutic agent against infections caused by methicillin-resistant S. aureus (MRSA). The main objective of this study was to examine the occurrence of S. aureus species in dental caries specimens. Antimicrobial susceptibility testing of S. aureus to four antibiotics namely vancomycin, linezolid, teicoplanin, and daptomycin was performed. Detection of vancomycin resistance was conducted using polymerase chain reaction. Among the tested 150 strains, 98 were MRSA and of that 54 were vancomycin sensitive and 27 were resistant. All 98 MRSA strains were positive for mecA and 36 yielded pvl, whereas 13 carried vanA and only 2 were positive for vanB. Majority of the isolates showed sensitivity toward daptomycin and linezolid. Strains of S. aureus exhibiting decreased susceptibility to different antibiotics like vancomycin, daptomycin, and linezolid severely compromise the therapeutic alternatives and require a considerable amount of time, public awareness, and integrative health-care strategies to prevent the emergence of resistance to these compounds. [ABSTRACT FROM AUTHOR]

Background. Vancomycin-resistant Enterococcus faecium (VREfm) is a leading cause of nosocomial infection. Here, we describe the utility of whole-genome sequencing in defining nosocomial VREfm transmission. Methods. A retrospective study at a single hospital in the United Kingdom identified 342 patients with E. faecium bloodstream infection over 7 years. Of these, 293 patients had a stored isolate and formed the basis for the study. The first stored isolate from each case was sequenced (200 VREfm [197 vanA, 2 vanB, and 1 isolate containing both vanA and vanB], 93 vancomycin-susceptible E. faecium) and epidemiological data were collected. Genomes were also available for E. faecium associated with bloodstream infections in 15 patients in neighboring hospitals, and 456 patients across the United Kingdom and Ireland. Results. The majority of infections in the 293 patients were hospital-acquired (n = 249) or healthcare-associated (n = 42). Phylogenetic analysis showed that 291 of 293 isolates resided in a hospital-associated clade that contained numerous discrete clusters of closely related isolates, indicative of multiple introductions into the hospital followed by clonal expansion associated with transmission. Fine-scale analysis of 6 exemplar phylogenetic clusters containing isolates from 93 patients (32%) identified complex transmission routes that spanned numerous wards and years, extending beyond the detection of conventional infection control. These contained both vancomycin-resistant and -susceptible isolates. We also identified closely related isolates from patients at Cambridge University Hospitals NHS Foundation Trust and regional and national hospitals, suggesting interhospital transmission. Conclusions. These findings provide important insights for infection control practice and signpost areas for interventions. We conclude that sequencing represents a powerful tool for the enhanced surveillance and control of nosocomial E. faecium transmission and infection. [ABSTRACT FROM AUTHOR]

Background Tigecycline is a new, semisynthetic glycylcycline. It is active against important multidrug resistant pathogens. Aim The purpose of this study was to investigate the sensitivity of multidrug-resistant enterococci to tigecycline, and to test the correlation between the minimal inhibitory concentration (MIC) and disk diffusion methods. Materials and methods The antimicrobial sensitivity of 108 multidrug-resistant isolates, which included 52 vancomycin-resistant enterococci (VRE) and 56 high-level aminoglycoside-resistant (HLAR) enterococci, was tested by the E test, broth microdilution test and disk diffusion methods. Results All of the isolates were sensitive to tigecycline, as determined by the E test and broth microdilution test. The MIC 90 value (0.19 μg/mL) of tigecycline for HLAR enterococci was higher than that for VRE (0.094 μg/mL). When results were evaluated according to species, the MIC values of tigecycline for Enterococcus faecalis were higher than those for the other species. Eleven (10.1%) isolates produced false resistance results (zone diameter ⩽15 mm) by the disk diffusion method. These cases were classified as major errors. Eight (7.4%) isolates had intermediate sensitivity (sensitivity zone of 16 or 17 mm), which were classified as minor errors. The major and minor error percentages of HLAR enterococci (14.2% major, 10.7% minor error) were higher than those of VRE (5.7% major, 3.8% minor error). These results indicate that tigecycline is effective against multidrug-resistant enterococci. The sensitivity of multidrug-resistant enterococci to tigecycline should be investigated by MIC methods. The disk diffusion method causes major errors, especially for HLAR enterococci. [ABSTRACT FROM AUTHOR]

The study was aimed to characterize enterococci from various clinical specimens, to determine the antimicrobial susceptibility pattern, and to explore the association between virulence factors and antimicrobial resistance. A total of 283 clinical enterococcal isolates were speciated and subjected to antimicrobial susceptibility testing. Virulence factors (hemolysin, gelatinase, and biofilm production) were detected phenotypically. Of the 283 enterococci isolated, 12 species were identified; predominant species were Enterococcus faecalis (82.33%). High-level gentamicin (HLG) and vancomycin resistance were observed among 55.57% and 6.01% of enteroccal isolates, respectively. All vancomycin-resistant enterococci (VREs) were E. faecalis and had VanA phenotype and genotype. Hemolysin, gelatinase, and biofilm production were seen in 15.90%, 12.36%, and 13.43% of enterococcal isolates, respectively. Vancomycin and HLG resistance were observed in 0.35% and 61.86% of the enterococcal isolates producing virulence factors. Isolates resistant to HLG but susceptible to vancomycin expressed more virulent factors. Further research is required to reveal the complex interplay between drug resistance and virulence factors. [ABSTRACT FROM AUTHOR]