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The unique properties of biosurfactants obtained from microbes, including their activity at extreme temperatures, make them more attractive than synthetic alternatives. Henceforth, the principle objective is to isolate and detect the antibacterial and antifungal activities of the biosurfactants produced from bacteria of the economically competitive mangrove ecosystem. Using the serial dilution method, 53 bacterial strains were recovered from the Manakudy mangrove forest in Kanyakumari, India, for the investigation. Different biosurfactant screening methods and morphological and biochemical tests were opted to select the potential biosurfactant producer. After the initial screening, two strains were discovered by 16S rRNA gene sequencing followed by extraction using chloroform: methanol (2:1) by the precipitation method. The partially purified biosurfactants were then screened for antimicrobial properties against pathogens including Mucor sp., Trichoderma sp. Morphological, biochemical, and 16S rRNA gene sequencing identified the two strains to be gram-positive, rod-shaped bacteria namely Virgibacillus halodentrificans CMST-ZI (GenBank Accession No.: OL336402.1) and Pseudomonas pseudoalcaligenes CMST-ZI (GenBank Accession No (10 K): OL308085.1). The two extracted biosurfactants viz., 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester, as well as cycloheptane efficiently inhibited human pathogens, including Enterococcus faecalis, and fungi, including Mucor sp., Trichoderma sp., indicated by the formation of a zone of inhibition in pharmacological screening. Thus, there is a growing interest in the prospective application of these biosurfactants isolated from marine microbes, exhibiting antimicrobial properties which can be further studied as a potential candidate in biomedical studies and eco-friendly novel drug development. [ABSTRACT FROM AUTHOR]

Introduction: Enterococci are major cause of health care associated infections and are resistant to commonly used antibiotics like cephalosporins. The study aims to find the proportion and characteristics of Enterococcus faecium isolates among the enterococcal infections at a tertiary care hospital. Methods and Material: A prospective cohort study was conducted in a tertiary care hospital in Mangalore, Karnataka for a period of 6 months from June to November 2023. The enterococci were isolated and identified and tested for antibiotic susceptibility by the standard methods. Results: Among the 132 enterococcal strains isolated from the clinical samples, 68 (51.51%) were E. faecalis and 61(46.21%) were E. faecium. E. faecium was significantly isolated more from the urine samples and from the inpatients. Also, the resistance to ampicillin (57.4% vs 14.7%), high level gentamicin (54.1% vs 36.8%), nitrofurantoin (37.7% vs 8.8%) and vancomycin (11.5% vs 1.5%) was significantly more among the E. faecium isolates, compared to the E. faecalis isolates. Resistance to linezolid was not found in the current study. Conclusion: In the present study, E. faecium strains were isolated in higher proportion compared to other studies. Therefore, there is an emergent need to combat this increase in E. faecium infections and the associated antibiotic resistance by scrutinising and encouraging appropriate antibiotic use in the hospitals and community. [ABSTRACT FROM AUTHOR]

Introduction: Blood culture is a proven gold standard method for the identification of causative agents of bloodstream infections. Prompt identification (ID) and antimicrobial susceptibility testing (AST) of organisms causing blood stream infections has a significant impact on the morbidity and mortality associated with these infections. The need to circumvent the slow turnaround time of conventional gold standard methods has paved way for the rapid automated systems. In this regard automated identification aid in rapid diagnosis/treatment of bacterial pathogens. Aims: Evaluation of automated system Vitek-2 compact system with conventional blood culture method for the identification of organisms causing neonatal septicaemia. Materials and Methods: In this hospital-based retrospective study conducted in the Department of Microbiology in a tertiary care hospital, New Delhi. A total of 170 non duplicate isolates of both Gram Positive and Gram-Negative organisms recovered from blood samples were included. Organisms were processed on the Vitek-2 compact system and simultaneously from conventional method (ID) for comparison. Descriptive statistics was used for the presentation and comparison of data and appropriate statistical charts were used to present the data. Results: Concordant identification (ID) results of Vitek-2 compact system were seen with all the isolates of S. aureus, S. epidermidis, S. pneumonia, E. faecalis and E. faecium. Discordant results of Vitek-2 were noticed with S. hominis (5 isolates of the organism misidentified as S. epidermidis). Conclusion: The organisms having slow metabolic rates and late lactose fermenters (S. hominis) are prone to errors by the Vitek-2 compact system; hence need to be reconfirmed with other possible method. [ABSTRACT FROM AUTHOR]

Background: Asymptomatic bacteriuria (ASB) during pregnancy can lead to symptomatic urinary tract infection (UTI), with increased fetal and maternal morbidity and mortality. We evaluated the incidence, clinical and microbiological profile, and outcome of ASB in pregnant women attending our antenatal clinic. Materials and Methods: This prospective study was conducted on 3769 pregnant women in a routine antenatal clinic at a tertiary care center. Participants were divided into two groups, ASB and non-bacteriuria. Data were collected in a standard proforma and analyzed using the software Statistical Package for the Social Sciences (SPSS) v. 20. Results: The incidence of ASB was 3.29% (124/3769). Majority of the women were in the age group of 21-30 years (78.76%, n = 89). Escherichia coli (61.06%) was the most common organism isolated. Maternal anemia (30.08% and 2.93% in the ASB and non-bacteriuria groups, respectively), low birth weight (LBW; 42.5% and 27.98% in the ASB and non-bacteriuria groups, respectively), intrauterine death (4.4% and 1.4% in the ASB and non-bacteriuria groups, respectively), and preterm delivery (37.2% and 22.31% in the ASB and non-bacteriuria groups, respectively) were were associated with ASB (P = 0.001). Conclusion: ASB was associated with maternal anemia, preterm delivery, intrauterine death, and LBW. Early detection and treatment of ASB may result in favorable maternal outcome. [ABSTRACT FROM AUTHOR]

Introduction: Enterococcus faecalis (E. faecalis) is a significant endodontic pathogen, and its virulence has been shown to be significantly influenced by Enterococcal fibronectin binding protein-A (EfbA). Targeting this virulent protein using alternative strategies would be a novel idea to combat the complications of E. faecalis in dental healthcare settings. Thus, the rationale of this investigation is to identify potent bioactive compounds from Aegle marmelos for their antimicrobial properties against EfbA of E. faecalis. Aim: To assess the frequency of EfbA among clinical isolates of E. faecalis and evaluate the antibacterial activity of essential bioactive compounds from Aegle marmelos. Materials and Methods: An in-vitro and in-silico pilot study was conducted from April 2022 to June 2022 in the Department of Microbiology, Saveetha Dental College and Hospitals, Chennai, Tamil Nadu, India to detect the frequency of EfbA in E. faecalis. Patients with typical root caries were included. Microbiological processing of 20 root caries samples was performed to characterise E. faecalis and detect the EfbA gene using Polymerase Chain Reaction (PCR) amplification. Crude methanol extract was obtained from A. marmelos and assessed for its antimicrobial effect against the isolated strains of E. faecalis. Furthermore, several biomolecules from A. marmelos were evaluated for their inhibitory properties through in-silico docking analysis. Results: E. faecalis was identified in 12 (60%) out of 20 root caries samples, and three isolates (25%) were identified as Multidrug-Resistant (MDR) strains based on the antibiogram susceptibility profile, showing resistance to more than three different tested antibiotic groups. EfbA was detected in two of the three MDR strains (66.7%). The crude extract of A. marmelos exhibited promising antibacterial activity. In-silico analysis of the essential oil compounds from A. marmelos revealed that Aegeline had a high interaction with low docking energy and a high number of hydrogen bonds. Conclusion: The current study highlights the potential of aegeline from A. marmelos as an antibacterial agent against resistant strains of E. faecalis. However, additional in-vivo research must be conducted to experimentally validate these findings. [ABSTRACT FROM AUTHOR]

Introduction: Multiple antibiotic resistances in the bacteria that cause infections in humans have been directly caused by the ongoing use of antibiotics in clinical practise. In undeveloped and impoverished nations, the overuse and inappropriate use of antibiotics may increase the risk of resistant bacteria emerging and proliferating throughout the population. Consequently, the antibiotics' effectiveness is declining. Therefore, there is now more interest in the therapeutic use of natural materials due to the need for innovative alternative antibacterial techniques. Aim and Objective: To study the antimicrobial activity of honey against clinical isolates of Methicillin-Resistant Staphylococcus aureus with special reference to Mec A gene, from various clinical samples at a tertiary care centre, Uttar Pradesh, India. Material and Methods: This was a prospective study carried out in the Department of Pharmacology and the Microbiology Department for a period of 1 year i.e, August 2022 to August 2023 at a Tertiary care centre, Uttar pradesh. The antibacterial activity of Dabur pasteurised honey was evaluated against the bacterial strains of Methicillin Resistant Staphylococcus aureus and Methicillin Sensitive Staphylococcus aureus. Their antibacterial sensitivity pattern was tested using Kirby-Bauer disc diffusion susceptibility testing technique according to the CLSI guidelines 2022 along with other commonly used antimicrobials. Results: A total of 200 clinical isolates were isolated, out of which 72 isolates were of S.aureus in which a total of 24 MRSA isolates were were identified by CX, OX, and E-test. The DNA was extracted using the DNA extraction Qiagen Kit and the Mec A gene was detected by the PCR. In the present study there were 24 isolates of MRSA out of which 15(62.5%) isolates were sensitive to honey and 48 isolates of (MSSA) Methicillin sensitive S. aureus out of which 42(87.5%) isolates were sensitive to honey. It was observed that all the Methicillin resistance isolates were resistant to Cefoxitin and Oxacillin whereas sensitive to linezolid, Teicoplanin and Vancomycin. In the present study the results of antibacterial activity of honey towards the two microorganisms tested were performed. MRSA as well as MSSA were sensitive to undiluted honey samples tested with an average zone of inhibition of 30.03± 0.1 and 41.06±0.2 respectively. Both MRSA and MSSA isolates were sensitive to honey. But MRSA were resistant to all antimicrobials tested except linezolid where as MSSA were sensitive to all except penicillin. Conclusion: Honey is undoubtedly a viable candidate for further research and testing as an antibacterial in the future. With more research on its mode of action at the molecular level, honey could find extensive application as an antibacterial agent in the future. [ABSTRACT FROM AUTHOR]

Introduction: The emergence of strains resistant to methicillin and other antimicrobials has become a major concern, particularly in the hospital settings as MRSA infections are resistant to the majority of conventional antibiotics, possesing serious risks to hospitals and communities all over the world. Aim and Objectives: To study the prevalence and the molecular characterization of Mec A gene in MRSA isolates at a tertiary care centre, Uttar Pradesh, India. Material and Methods: This was a cross sectional study conducted in the Department of Microbiology at a tertiary care centre, for a period of 1 year i.e, August 2022 to August 2023. A total of 682 clinical isolates was studied out of which 200 isolates of S. aureus were identified using the biochemical test from the clinical samples such as pus, swab, blood, wound and urine etc. The different Phenotypic Methods including Cefoxitin and Oxacillin Disc Diffusion test and the genotypic method including MecA gene detection for Methicillin Resistant Staphylococcus aureus isolates was performed. The DNA was extracted using the DNA extraction Qiagen Kit and the Mec A gene was detected by the PCR method. Results: In the present study a total of 42 MRSA isolates were identified by CX, OX, and E-test test phenotypically. The prevalence of MRSA was found to 21%. The maximum number of isolates were observed from the OPD ward with 38.5%. The ratio of the males 28 (66.6%) was more as compared to the females with 14 (33.3%) with the maximum age of 41-50 years being affected the most followed by 31-40 years and least in the age group of 61 years and above. The pus 47.6% was the most common isolate followed by the blood with 23.8% and least for urine and the body fluids 2.3%. All the MRSA isolates were found sensitive to linezolid, Teicoplanin, vancomycin, however all the isolates were recorded resistant with Cefoxitin and Oxacillin. The presence of MecA gene was recorded in all the 42 isolates of MRSA. The presence of MecA gene was confirmed by the PCR followed by sequencing assay. Conclusion: This study provides a clear guidance to effectively diagnose and measure MRSA infections in hospitals and communities. Future research could identify MRSA isolates with the help of the data produced in this investigation. For the control of antibiotic resistance, there is a need for ongoing monitoring and the deployment of effective control techniques where continuous surveillance, awareness of the incidence of MRSA, and upkeep of hygienic standards have been needed to reduce MRSA infections. [ABSTRACT FROM AUTHOR]

Introduction Gram-positive bloodstream infections (BSIs) are an emerging health concern, especially in resource-limited settings. There is a paucity of data regarding the antimicrobial resistance (AMR) pattern of Gram-positive BSIs. The rise in multidrug-resistant infections further convoluted antibiotic selection. We aimed to assess the incidence, clinical and microbiological profile, antimicrobial resistance (AMR) and outcome in Gram-positive BSIs. Methods This was a single-center prospective study conducted at a tertiary care hospital in Western India. All patients (age ≥18 years) with culture-proven Gram-positive BSIs were included. Data were collected on all patients' demography, risk factors, AMR and clinical outcome. Results A total of 210 clinically significant isolates were grown from July 2020 to December 2021. The incidence of Gram-positive BSIs was 29% (n=61); 55.9% of cases were healthcare-associated, while 44.1% were community-acquired. Coagulase-negative staphylococci (CoNS) were the major isolates (36.1%), followed by Enterococcus spp. (27.9%), methicillin-susceptible Staphylococcus aureus (MSSA) (18%) and methicillin-resistant Staphylococcus aureus (MRSA) (14.7%). The proportion of vancomycin and teicoplanin-resistant CoNS isolates was 13.6% and 19%. Among Enterococcus isolates, the proportion of vancomycin-resistant enterococci (VRE) and linezolid-resistant enterococci (LRE) were 11.8% and 5.9%. The overall mortality in Gram-positive BSIs was 42.6%. Older age, MRSA infection, septic shock, and high NLR were significantly associated with mortality. On the Cox regression model, age ≥65 years (HR: 2.5; 95%CI: 1.1-5.8; p=0.024) and MRSA infection (HR: 3.6; 95%CI: 1.5-8.5; p=0.021) were found as independent predictors of 30-day mortality. Conclusions This study found substantial mortality with Gram-positive BSIs, especially MRSA infections. Moreover, the emergence of VRE and LRE is also alarming. Active surveillance of AMR and evaluation of mortality predictors may help overcome the therapeutic challenges in managing BSIs. [ABSTRACT FROM AUTHOR]

Background: Vancomycin-resistant Enterococci (VRE) infections are recurrently reported in different parts of India in the last two decades. However, an up-to-date, countrywide information concerning the prevalence and the rate of VRE in India is limited and hence this study aimed to estimate the pooled prevalence of VRE in India. Methods: A literature search was performed using various databases. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed throughout. Cross-sectional studies reporting the prevalence of VRE in India from human samples whereby at least two Enterococci were isolated between 1 January 2000 and 31 December 2022 were sought for inclusion. Data were extracted and analysed using Microsoft Excel and Comprehensive Meta-analysis version 4, respectively. Results: Nineteen studies were included in the analyses. A collective total of 3683 Enterococci isolates were examined, of which 368 were VRE strains. The pooled prevalence of VRE in India was calculated at 12.4% (95% CI: 8.6–17.5; Q = 189.69; I2 = 90.51%; p = < 0.001). E. faecalis was the most frequently isolated species (1450 [39.37%]) followed by E. faecium (724 [19.66%]). Amongst the VRE strains, E. faecium was the most prevalent (214 [58.15%]) followed by E. faecalis (134 [36.41%]). An upsurge in the rate of VRE infections was observed in India over time: VRE prevalence was estimated at 4.8% between 2000 and 2010 and 14.1% between 2011 and 2020. Conclusion: This study presents the most up-to-date information on the rate of VRE infections in India. Though lower than the findings for some less developed countries, VRE prevalence in India is notable and on the rise. [ABSTRACT FROM AUTHOR]

Background and Aims: Urinary tract infections are one of the most prevalent illnesses with high morbidity and mortality rates (UTIs). Despite the availability of numerous medications for the treatment of UTIs, uropathogens have developed resistance, resulting in a major treatment failure. This experiment was done to determine the rate of pathogen isolation and the pattern of antibiotic susceptibility of microorganisms causing urinary tract infections. Materials and procedures: Between November and January of 2023, 250 urine samples from people with a suspected UTI were processed. Using the Kirby-Bauer disc diffusion method, the susceptibility of several drugs and UTI isolates was examined. Results: Among the 450 samples analyzed in this investigation, 165 (56%) exhibited microbial growth. 23 of the 165 isolates were Candida species, while 142 were bacteria. The predominant isolate was E. coli (47%), followed by E. faecalis (24%) and Klebsiella spp. (20%). Candida spp was most susceptible to Fluconazole (91%), while E. coli was most susceptible to nitrofurantoin (97%). The results of the study demonstrated that E. coli is the most prevalent uropathogen that causes UTIs, and that Nitrofurantoin is the superior antibiotic with a high susceptibility rate. [ABSTRACT FROM AUTHOR]

Fermented food and beverages have potential novel Probiotic microorganism which possess many beneficial role in human health and hygiene. The tribal people consume variety of locally produced fermented food and beverages regularly and hence their gut microflora are strongly benefited. Mayurbhanj district in Odisha, India is diversified with many tribal population. Handia, is the most preferred fermented rice beverage in the tribal community having cultural, religious and social values associated with it. Recent advances in the fields of science like nanoscience and nanotechnology have radically changed the way we diagnose, treat, and prevent various diseases in all aspects of human life. This current investigation was undertaken to explore the various possibilities to screen the potentiality of utilizing Enterococcus lactis, a probiotic bacteria isolated from the fermented rice beverage "Handia" in terms of their efficacy for synthesis of Silver nanoparticles (AgNPs). The synthesized AgNPs were screened through Fe-SEM, EDS, XRD, FTIR, DLS and UV-visible spectroscopy characterization studies. The synthesized AgNPs exhibited enhanced antioxidant, antimicrobial and anti-biofilm forming capability. Understanding the implications of green-synthesized AgNPs from probiotic bacteria would help in designing and delivery of bioactive components present in probiotic microorganism for future drug delivery application and other nanoparticle based therapeutics in different sectors of biomedical engineering and technology. [ABSTRACT FROM AUTHOR]

Background: There are no eradication approaches for Vancomycin Resistant Enterococci(VRE) so infection control strategies rely only on screening and isolation of colonised patients. Objective: To find the prevalence and distribution of Vancomycin Resistant Enterococci(VRE). Methodology: This was an observational study from a tertiary care hospital which was completed over a period of 18 months. Results: The prevalence of Enterococcal isolates was3.9% in our hospital out of which 19.6% were Vancomycin Resistant Enterococci(VRE). Among male subjects the prevalence was much higher (27.5%)in comparison to females(12%) and the difference was found to be statistically significant (p < 0.001). Conclusion: This study emphasizes on the prevalence of vancomycin resistant Enterococci in Tertiary care hospital of Kashmir valley. Enterococcus species has been of particular concern due to their increasing incidence and paucity of drugs available to treat them. [ABSTRACT FROM AUTHOR]

BackgroundStaphylococcus haemolyticus has emerged as an important multidrug-resistant nosocomial pathogen. Linezolid is useful in the treatment of severe infections caused by methicillin-resistant Staphylococci. Resistance to linezolid in Staphylococci is due to one or more of the following mechanisms: acquisition of the cfr (chloramphenicol florfenicol resistance) gene, mutation in the central loop of domain V of the 23S rRNA, and mutation in the rplC and rplD genes. This study was carried out to detect and characterize resistance to linezolid among the clinical isolates of Staphylococcus haemolyticus. Materials and Methods The study included 84 clinical isolates of Staphylococcus haemolyticus. Susceptibility to various antibiotics was determined by disc diffusion method. Minimum inhibitory concentration (MIC) was determined by agar dilution method for linezolid. Methicillin resistance was screened using oxacillin and cefoxitin disc. Polymerase chain reaction was done to detect mecA, cfr and mutations in the V domain of the 23S rRNA gene. Results Resistance to linezolid was exhibited by 3 of the 84 study isolates with MIC more than 128 µg/mL. The cfr gene was detected in all the three isolates. The G2603T mutation was observed in the domain V of the 23S rRNA among two isolates, whereas one isolate lacked any mutation. Conclusion The emergence and spread of linezolid-resistant Staphylococcus haemolyticus isolates carrying G2603T mutation in the domain V of the 23S rRNA and harboring the cfr gene pose a threat in clinical practice. [ABSTRACT FROM AUTHOR]

Introduction: There is increasing development of antibiotic resistance among the Enterococcus species. Objectives: This study was performed to determine prevalence and characterize the vancomycin-resistant and linezolid-resistant enterococcus isolates from a tertiary care center. Moreover, the antimicrobial susceptibility pattern of these isolates was also determined. Materials and Methods: A prospective study was performed in Medical College, Kolkata, India, over a period of two years (from January 2018 to December 2019). After obtaining clearance from the Institutional Ethics Committee, Enterococcus isolates from various samples were included in the present investigation. In addition to the various conventional biochemical tests, the VITEK 2 Compact system was used to identify the Enterococcus species. The isolates were tested for antimicrobial susceptibility to different antibiotics using the Kirby–Bauer disk diffusion method and VITEK 2 Compact to determine the minimum inhibitory concentration (MIC). The Clinical and Laboratory Standards Institute (CLSI) 2017 guidelines were used to interpret susceptibility. Multiplex PCR was performed for genetic characterization of the vancomycin-resistant Enterococcus isolates and sequencing was performed for characterization of the linezolid-resistant Enterococcus isolates. Results: During the period of two years, 371 isolates of Enterococcus spp. were obtained from 4934 clinical isolates showing a prevalence of 7.52%. Among these isolates, 239 (64.42%) were Enterococcus faecalis, 114 (30.72%) Enterococcus faecium, and others were Enterococcus durans, Enterococcus casseliflavus, Enterococcus gallinarum, and Enterococcus avium. Among these, 24 (6.47%) were VRE (Vancomycin-Resistant Enterococcus) of which 18 isolates were Van A type and six isolates of Enterococcus casseliflavus and Enterococcus gallinarum were resistant VanC type. There were two linezolid-resistant Enterococcus, and they were found to have the G2576T mutation. Among the 371 isolates, 252 (67.92%) were multi-drug resistant. Conclusion: This study found an increasing prevalence of vancomycin-resistant Enterococcus isolates. There is also an alarming prevalence of multidrug resistance among these isolates. [ABSTRACT FROM AUTHOR]

The term 'toddy' represents a group of different varieties of mild-alcoholic palm beverages of coastal and inland India, produced from the fresh saps of various palm trees through uncontrolled natural fermentation. In this study, we analysed the successional changes of microbial abundances and various physico-chemical parameters during natural fermentation (0 h to 48 h) of taal toddy, prepared from Palmyra palm, and khejur toddy, prepared from date palm of West Bengal and Jharkhand in India. Microorganisms from different successional levels were isolated and grouped using repetitive element sequence-based PCR (rep-PCR) technique and identified by the sequencing of 16S rRNA gene and D1-D2 region of 26S rRNA gene for bacteria and yeasts, respectively. Enterococcus faecalis, Lactiplantibacillus plantarum, Lacticaseibacillus paracasei and yeast Saccharomyces cerevisiae were identified during natural fermentation of toddy. During the natural fermentation, the average pH and total sugar content in the samples of both taal and khejur toddy decreased, whereas a gradual rise was observed in the contents of acidity, total alcohol, total ester and total protein. Bio-active potential (presence of phenolics and flavonoids) of toddy was also analysed (0 h to 48 h), where contents of total phenolics, flavonoids and resulting anti-oxidant activity were found higher in the end-product than the fresh palm sap, indicating toddy as a functional low-alcoholic drink. Lastly, it can be concluded that the inter-variable dynamics and microbial interrelation, which in turn depend on a number of local factors, regulate the overall fermentation dynamics and determine the product quality. [ABSTRACT FROM AUTHOR]

Background and Aim: Vancomycin resistant Enterococci (VRE) and high level aminoglycoside resistant (HLAR) Enterococci have complicated the available treatment modalities for Enterococci worldwide. The existing study was planned to evaluate the occurrence of HLAR and VRE strains in a tertiary care center in India and to study the association of HLAR with vancomycin sensitive Enterococci (VSE) and VRE. Materials and Methods: A total of 50 enterococcal isolates from various clinical specimens were incorporated in the study. Speciation was done on the basis of standard biochemical tests. HLAR was tested by the disc diffusion method using 150μg gentamicin disc and 200 μg streptomycin discs. Vancomycin susceptibility patterns were reported using vancomycin disc and agar dilution methods. Results & Conclusion: Pus samples comprised of the majority for the isolation of enterococcal strains (40%). 54% isolated strains were HLGR, 32% were HLSR and 14% isolates were positive for both HLGR and HLSR. 61.7% of Enterococcus faecium isolates demonstrated resistance for high-level gentamicin (HLGR) and 43.75% Enterococcus faecalis isolates were resistant to high-level streptomycin (HLSR). When VRE was compared to VSE, the rate of HLSR was detected to be 4.64% in VRE, while it was 32.55% in VSE; the rate of HLGR was noted to be 11.62% in VRE and it was 41.87% in VSE. The association of HLGR with HLSR (HLAR) was 2.32% in VRE and 13.95% in VSE strains. Enterococci strains are showing an increase in their antimicrobial resistance patterns. The increment of such strains in health care settings has to be reserved and controlled to avert complicated infections. [ABSTRACT FROM AUTHOR]

Objective We aimed to evaluate the current antimicrobial susceptibility pattern and characterize putative virulence traits among Enterococcus species isolates from various clinical specimens in view of their increased isolation rates in both community-related and serious nosocomial infections, as well as resistance to many antibiotics. Methods Study (April 2017–March 2018) included consecutive, nonrepeated, discrete, and clinically significant isolates of enterococci. Susceptibility testing included detection of high-level aminoglycoside-resistant (HLAR) and glycopeptide-resistant enterococci (GRE). All screen-positive GRE isolates were investigated by polymerase chain reaction for species confirmation and presence of vanA/vanB genes. Virulence genes ace , asa1 , cyt , efa , esp , gelE , and hyl were investigated by molecular methods. Hemolysin and biofilm production were studied using phenotypic methods. Results Of 111 isolates, 89 (80.1%), 16 (14.4%), and 6 (5.4%) were from urine, pus, and blood, respectively, consisting predominantly of E. faecalis (67, 60.4%) and E. faecium (32, 28.8%). E. hirae (5, 4.5%) was the predominant non- E. faecalis non- E. faecium isolate. Other species were E. durans (4, 3.6%), E. avium (2, 1.8%), and E. mundtii (1, 0.9%). Seven (6.3%) out of the 111 isolates were GRE, all vanA genotype. HLAR was observed in 70 (63.1%) isolates, significantly higher in E. faecium than E. faecalis (81.2 vs. 58.2%; p < 0.05). All were susceptible to daptomycin. Hemolysin activity and biofilm production were observed in 38 (34.2%) and 36 (32.4%) isolates. Most frequent virulence genes were efa (77, 69.4%), ace (71, 63.9%), asa1 (67, 60.3%), and gelE (66, 59.4%). There was a predominant association of esp and hyl genes with E. faecium and that of the other genes with E. faecalis. Conclusion The study will contribute to the existing limited data on virulence trait characterization of clinical E. spp. isolates in India. At the same time, it will help to serve as a guide in the choice of empirical therapy in enterococcal infections leading to favorable clinical outcomes by decreasing the clinical failure, microbiological persistence, and associated mortality, and will lead to future studies on controlling the spread of virulent and multiresistant isolates. [ABSTRACT FROM AUTHOR]

Background: The emergence of Enterococcus as an important nosocomial pathogen is mainly attributed to its inherent resistance to commonly used antibiotics and now in recent times, it has acquired resistance to other available therapeutic options as well. Materials and Methods: Enterococcus isolates from clinical samples received in the department of microbiology over a period of 1 year were included in the study. Isolates were identified and species determined by standard methods. Antibiotic susceptibility test was done by Kirby Bauer disc diffusion test. Epsilometer test was done to determine the minimum inhibitory concentration for vancomycin. Results: A total of 145 Enterococcus isolates were obtained; 73 (50.3%) isolates were identified as Enterococcus faecalis, 69 (47.5%) Enterococcus faecium, 2 (1.3%) Enterococcus durans and 1 (0.68%) as Enterococcus gallinarum. Most of the isolates were from urine samples (125, 86.2%); followed by pus (16, 11.03%). Vancomycin resistance was seen in 14 (9.6%) isolates while linezolid resistance was seen in 8 (5.5%) isolates. Conclusion: E. faecalis is the most common clinical species isolated from clinical samples and the emergence of linezolid-resistant enterococci from the hospital is a matter of concern as till now it is considered to be the last resort for treatment in patients infected with vancomycin-resistant enterococci. [ABSTRACT FROM AUTHOR]

CONTEXT: Enterococci are known to cause life-threatening infections which are difficult to treat as the organism harbors innate resistance to many antibiotics and can amass resistance toward many others through plasmid-mediated genetic exchange. AIMS: The study evaluates the drug susceptibility profile of various Enterococcus species isolated from various patient specimens submitted for bacteriological analysis and check the incidence of aac(6′) Ie-aph(2") Ia gene. SETTING AND DESIGN: This in vitro cross-sectional study was executed at bacteriology laboratory of a 470 bedded hospital in New Delhi. MATERIALS AND METHODS: Drug susceptibility testing was carried out on enterococcal isolates. High-level gentamicin-resistant (HLGR) isolates detected by micro broth dilution assay were then subjected to molecular detection of aac(6′) Ie-aph(2") Ia gene. STATISTICAL ANALYSIS USED: The level of significance was established by Chi-square test. RESULTS: Among the 182 enterococcal stains detected, 76.9% were Enterococcus faecalis and 20.3% were Enterococcus faecium. 12.08% strains were vancomycin resistant. 39% expressed resistance toward high-level gentamicin (HLG) and this finding was significantly higher in E. faecium than E. faecalis. HLGR strains expressed a higher degree of resistance to other drugs in contrast to non-HLGR isolates. In 67 out of 71 HLGR isolates the bifunctional gene was detected. CONCLUSION: Considerable presence of HLG and vancomycin resistance in the clinical isolates is alarming and should be taken seriously. The study shows high dissemination of aac(6′)-Ie-aph(2")-Ia gene among Enterococci isolated from the region. [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]

A total of 272 isolates of lactic acid bacteria (LAB) were isolated from 22 samples of naturally fermented milk products of Sikkim in India viz. dahi, soft-variety chhurpi, hard-variety chhurpi, mohi and philu, out of which, 68 LAB isolates were randomly grouped on the basis of phenotypic characteristics, and were identified by 16S rRNA gene sequence analysis. Leuconostoc mesenteroides was the most dominant genus, followed by Leuc. mesenteroides subsp. jonggajibkimchii, Lactococcus lactis subsp. cremoris, Lc. lactis, Lc. lactis subsp. hordniae, Lc. lactis subsp. tructae, Enterococcus faecalis, E. italicus and E. pseudoavium. LAB strains were tested for probiotics attributes by in vitro and genetic screening, based on marker genes. LAB strains showed tolerance to pH 3.0, bile salt, resistance to lysozyme and β-galactosidase activity. Enterococcus faecalis YS4-11 and YS4-14 and Lactococcus lactis subsp. cremoris SC3 showed more than 85% of hydrophobicity. Genes clp L and tdc encoding for low pH tolerance, agu A and Ir1516 encoding for bile tolerance, LBA1446 gene encoding for BSH activity, map A, apf, mub 1 and msa encoding for mucosal binding property were detected. Gene mesY for bacteriocin production was detected only in Leuconostoc spp. Based on the in vitro and genetic screening of probiotic attributes, Leuc. mesenteroides; Leuc. mesenteroides subsp. jonggajibkimchii and Lc. lactis subsp. cremoris were tentatively selected for possible probiotic candidates. [ABSTRACT FROM AUTHOR]

Introduction: Vancomycin-resistant enterococci (VRE) are emerging as an important multidrug-resistant pathogen causing nosocomial infections, predominantly bacteremia and urinary tract infections. VRE bacteremia has caused a significant increase in the duration of the hospital stay and mortality and had caused high public health threat due to limited treatment options. Materials and methods: Between October 2017 and September 2020, all consecutive patients with culture-proven bloodstream infection with Enterococcus species, isolated for the first time, were included in the study. A total of 427 Enterococcus species were identified, and antimicrobial susceptibility tests were performed and interpreted using Clinical and Laboratory Standard Institute guidelines. Results: Of the total 427 Enterococcus species isolated, 63 (45.6%) were VRE. Among them, 51/63 (81%) were Enterococcus faecium (E. faecium) and 5/63 (8%) were Enterococcus faecalis. There was an increased trend of VRE rate in the bloodstream infections of 6.12% (2018), 13.2% (2019), and 19.2% (2020). The majority of the VRE patients [43/63 (68%)] were admitted to the intensive care units (ICUs). Vancomycin A (VanA) is the most common phenotype isolated from 51/63(81%) patients. Conclusion: This increasing trend of VRE bacteremia is a red alert to the clinicians and the infection control practitioners, so that strict antibiotic policies and proper adherence to the infection control practices can be initiated to reduce the VRE rate. [ABSTRACT FROM AUTHOR]

Background: Mastitis is a multi-etiological disease. It leads to physical, chemical and bacteriological changes in milk and pathological changes in glandular tissues. In the present study, prevalence of mastitis causing organisms with special reference to Pseudomonas aeruginosa and its antibiotic resistance pattern was studied as Pseudomonas spp. is an important organism in harbouring and transmission of antibiotic resistance in environment. Methods: One hundred and one milk samples from mastitic cattle and buffaloes from farms in and around Ludhiana and mastitis cases presented in veterinary clinical complex, GADVASU, Ludhiana, Punjab, India were collected from August 2018 till April 2019. All the samples were processed for isolation of bacteria using different media viz., BHI, EMB, MLA, Cetrimide Agar, BP Agar, Edward’s medium and Blood Agar. After the isolation, all the organisms were subjected to cultural, morphological, biochemical tests and PCR for confirmation. All these isolates (except Bacillus spp. and Serratia marcescens) were screened for their antimicrobial resistance against 15 commonly used antibiotics viz., chloramphenicol, erythromycin, tetracycline, amoxycillin, co-trimoxazole, ciprofloxacin, gentamicin, cephalexin, ofloxacin, sparfloxacin, gatifloxacin, teicoplanin, azithromycin, vancomycin and doxycycline. Result: Out of a total of 101 samples, 86 samples yielded bacterial growth. Out of these 86 samples, 76 (88.37%) samples had a single bacterial growth whereas rest 10 (11.63%) samples had more than one bacterial growth. It was observed after the organisms were subjected to cultural, morphological, biochemical tests and PCR that, a total of 33 (34.02%) Staphylococcus aureus, 24 (24.74%) Escherichia coli, 16 (16.49%) Klebsiella pneumoniae, 5 (5.15%) Pseudomonas aeruginosa, 2 (2.06%) Streptococcus agalactiae, 12 (12.37%) Bacillus spp. and Serratia marcescens 5 (5.15%) were isolated. It was observed that Pseudomonas aeruginosa showed resistance against most of the antibiotics and was sensitive only for ofloxacin (80%) and gentamicin (40%). [ABSTRACT FROM AUTHOR]

• India: a densely populated country with evolving antimicrobial stewardship. • 2010-20: increased antibiotic resistance of Gram-negative ESKAPE in India. • 2010-20: increased resistance against [β-lactams/β-lactamase inhibitors] in India. • India: required intensified antimicrobial stewardship to curb ESKAPE resistance. • India: urgent need for continuous monitoring and documenting antimicrobial resistance. ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) constitute a threat to humans worldwide. India is now the most populous country. The goal was to investigate the evolution of the rates of antimicrobial resistance in ESKAPE pathogens across India over the 2010-20 decade. The data (89 studies) were retrieved from the Medline PubMed repository using specific keywords. The study of 20 177 ESKAPE isolates showed that A. baumannii isolates were the most represented (35.9%, n = 7238), followed by P. aeruginosa (25.3%, n = 5113), K. pneumoniae (19.5%, n = 3934), S. aureus (16.3%, n = 3286), E. faecium (2.6%, n = 517) and Enterobacter spp. (0.4%, n = 89). A notable increase in the resistance rates to antimicrobial agents occurred over the 2010-20 decade. The most important levels of resistance were observed in 2016-20 for A. baumannii (90% of resistance to the amoxicillin-clavulanate combination) and K. pneumoniae (81.6% of resistance to gentamycin). The rise in β-lactamase activities was correlated with an increase in the positivity of Gram-negative isolates for β-lactamase genes. This review highlighted that, in contrast to developed countries that kept resistance levels under control, a considerable increase in resistance to various classes of antibiotics occurred in ESKAPE pathogens in India over the 2010-2020 decade. [ABSTRACT FROM AUTHOR]

Introduction: Enterococci are important agents of nosocomial infection, ranking as the second most common organisms causing complicated urinary tract infections, bacteraemia, endocarditis, intra-abdominal and pelvic infections, wound and soft tissue infections, neonatal sepsis, and, rarely, meningitis. Infections by enterococci have traditionally been treated with cell wall active agents (e.g., penicillin or ampicillin) in combination with an aminoglycoside (streptomycin/gentamicin); however, emergence of High Level Aminoglycoside Resistance (HLAR), ß lactam antibiotics resistance and vancomycin resistance by some strains has led to failure of synergistic effects of combination therapy. Aim: To characterise enterococci up to the species level and study their antibiotic susceptibility pattern. Materials and Methods: The present study was a crosssectional study in which a total of 14114 clinical specimens, obtained during the period from September 2018 to August 2020 in this cross-sectional study, were tested to identify and speciate enterococcal isolates using standard microbiological methodology. Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method. Data was analysed using descriptive statistics (percentage and proportion). Results: During the study period of two years, 146 enterococci were recovered from 14114 different clinical samples, accounting for an infection rate of 1.03%. Among 146 enterococcal isolates, 116 (79.5%) were obtained from urine, 13 (8.9%) from blood, 10 (6.8%) from pus, 4 (2.7%) from wound swab and 3 (2.1%) from catheter tip. The predominant isolates were E. faecalis (82.2%) followed by E. faecium (15.8%), E.durans (1.3%) and E.gallinarum (0.7%). On studying the antibiotic susceptibility pattern, most of enterococcal isolates were predominantly resistant to ampicillin and ciprofloxacin (73.9% in both) and least resistant to linezolid (3.4%). Conclusion: Enterococcus faecalis and Enterococcus faecium were the predominant species in present study and majority of the isolates was sensitive to linezolid (96.6%). Therefore, it is necessary to implement infection control measures like antimicrobial stewardship especially restricting the use of antibiotics to minimum. [ABSTRACT FROM AUTHOR]

Introduction: Tigecycline is a potential therapeutic agent for multidrug resistant gram positive and gram negative organisms. The clinical efficacy of synergic combination of daptomycin with betalactam antibiotics have been described against various species of Enterococcus isolates. Aim: To detect the in-vitro activity of tigecycline and daptomycin against Enterococcus species and to compare their antimicrobial activity by Vitek 2 automated system. Materials and Methods: This was a cross-sectional study conducted at Department of Microbiology, Vinayaka Mission's Kirupananda Variyar (VMKV) Medical College and Hospitals, Salem, Tamil Nadu, India. Isolates of Enterococcus obtained over a period of two years from March 2016 - March 2018 from various clinical samples were identified by standard biochemical method and their antimicrobial susceptibility pattern was determined. Minimum Inhibitory Concentration (MIC) of tigecycline and daptomycin was determined by Vitek 2 automated system. Analysis of data was done by using frequency distribution and percentage. Results: Out of 211 Enterococcus isolates studied, 23 (10.90%) isolates showed decreased susceptibility to daptomycin with MIC of =8 µg/mL. All isolates showed 100% susceptibility to tigecycline. Conclusion: The present study showed potent antimicrobial activity of tigecycline against various species of Enterococcus. Decreased susceptibility to daptomycin has been observed among the study isolates. Further clinical investigations are required to know the potential benefits of these agents in therapy. [ABSTRACT FROM AUTHOR]

With the increasing emergence of drug resistance in enterococci, there have been very limited data on the efficacy of orally available nitrofurantoin and fosfomycin on enterococci causing urinary tract infections (UTIs), particularly for multidrug-resistant (MDR) strains. This study aimed to determine the in vitro effectiveness of these two drugs against the MDR enterococci. A total of 514 phenotypically and genotypically confirmed isolates of enterococci (239, 46.5% Enterococcus faecalis and 275, 53.5% Enterococcus faecium) showed E. faecalis as significantly more resistant (p < 0.05) to ciprofloxacin and high strength gentamicin. Vancomycin resistance was seen in 37 (7.2%) isolates. Of these, 114 (22.18%) isolates (51, 44.73% E. faecalis and 63, 55.26% E. faecium) were MDR. Nitrofurantoin minimum inhibitory concentrations (MICs) for the MDR enterococci varied from 1 to 128 μg/mL (MIC50 8 μg/mL, MIC90 64 μg/mL for E. faecalis), while fosfomycin MICs for the MDR E. faecalis, including vancomycin resistant enterococci (VRE) were in susceptible range (≤64 μg/mL, MIC50 8 μg/mL, MIC90 16 μg/mL). An efficacy ratio of ≥8 for nitrofurantoin was observed in the 39 (76.5%) MDR E. faecalis and 44 (69.8%) MDR E. faecium isolates as against the 50 (98%) E. faecalis isolates for fosfomycin. Although nitrofurantoin has been widely prescribed for the treatment of UTIs for the past several years, it was still found to be active in vitro against the urinary isolates of MDR enterococci, including VRE. As for fosfomycin, it holds robust potential to be used against the urinary MDR enterococci and VRE (E. faecalis). [ABSTRACT FROM AUTHOR]

Soil salinization has become a prominent obstacle in diverse arid and semi-arid region damaging agricultural productivity globally. From this perspective, present investigation was aimed to compare the potential compatible consortium of bio-inoculants for improving Plant Growth Promoting (PGP) attributes, anti-oxidative enzymes, grain yield and profitability of Vigna radiata in saline soil conditions. A total of 101 rhizobacterium isolated from salt affected regions of Punjab, India were screened for their ability to induce salt tolerance, multifunctional PGP traits and antagonistic activities. The 16S rRNA sequencing identified the strains LSMR-29 and LSMRS-7 as Pseudomonas flourescens and Enterococcus hirae , respectively. In-vitro compatible halo-tolerant dual inoculant (LSMR-29 + LSMRS-7) as bio-inoculants mitigated salt stress in Vigna radiata (spring mungbean) seedling with improved seed germination, biomass and salt tolerance index together with the presence of nif H, acds , pqq and ipdc gene under salinity stress as compared to single inoculants. Further, the potential of single and dual bio-inoculants were also exploited for PGP attributes in pot and field experiments. Results indicated that a significant improvement in chlorophyll content (2.03 fold), nodulation (1.24 fold), nodule biomass (1.23 fold) and leghemoglobin content (1.13 fold) with dual inoculant of LSMR-29 + LSMRS-7 over the LSMR-29 alone. The concentrations of macro & micronutrients, proline, soil enzyme activities i.e. soil dehydrogenase, acid & alkaline phosphatases and antioxidant enzymes such as superoxide dismutase, catalase and peroxidase also found to be high for LSMR-29 + LSMRS-7 as compared to un-inoculated control. The high grain yield thereby leading to Benefit: Cost (B: C) ratio at field scale was indicative of the commercial use bio-inoculants under salt affected Vigna radiata (spring mungbean) to improvement of productivity and soil health. The current finding reveals a co-inoculation of halo-tolerating Pseudomonas fluorescens and Enterococcus hirae containing ACC deaminase could prove to be novel approach for inducing salt tolerance and improving productivity of Vigna radiata (spring mungbean). [Display omitted] • Potential strains LSMR-29 & LSMRS-7 increased abiotic tolerance and colonization abilities. • Pseudomonas fluorescens and Enterococcus hirae employed as bio-stimulants. • Multifunctional PGP activities mitigate agricultural stress were summarized. • Using bio-stimulants is a sustainable and promising approach to enhance crop production. • It is novel strategy for inducing salt tolerance, improving yield and soil health. [ABSTRACT FROM AUTHOR]

Grep-chhurpi, peha, peron namsing and peruñyaan are lesser-known home-made fermented soybean foods prepared by the native people of Arunachal Pradesh in India. Present work aims to study the microbiome, their functional annotations, metabolites and recovery of metagenome-assembled genomes (MAGs) in these four fermented soybean foods. Metagenomes revealed the dominance of bacteria (97.80 %) with minor traces of viruses, eukaryotes and archaea. Bacillota is the most abundant phylum with Bacillus subtilis as the abundant species. Metagenome also revealed the abundance of lactic acid bacteria such as Enterococcus casseliflavus, Enterococcus faecium, Mammaliicoccus sciuri and Staphylococcus saprophyticus in all samples. B. subtilis was the major species found in all products. Predictive metabolic pathways showed the abundance of genes associated with metabolisms. Metabolomics analysis revealed both targeted and untargeted metabolites, which suggested their role in flavour development and therapeutic properties. High-quality MAGs, identified as B. subtilis, Enterococcus faecalis, Pediococcus acidilactici and B. velezensis, showed the presence of several biomarkers corresponding to various bio-functional properties. Gene clusters of secondary metabolites (antimicrobial peptides) and CRISPR-Cas systems were detected in all MAGs. This present work also provides key elements related to the cultivability of identified species of MAGs for future use as starter cultures in fermented soybean food product development. Additionally, comparison of microbiome and metabolites of grep-chhurpi, peron namsing and peruñyaan with that of other fermented soybean foods of Asia revealed a distinct difference. • Some spontaneously fermented soybean foods of India were studied using metagenomics and metabolomics approaches. • Bacillus subtilis as the abundant species, few lactic acid bacteria, and coagulase-negative staphylococci. • Untargeted metabolites including flavour compounds, isoflavones, immunomodulators and bio-peptides detected in samples. • High-quality MAGs B. subtilis, B. velezensis Enterococcus faecalis, Pediococcus acidilactici recovered with biomarkers. [ABSTRACT FROM AUTHOR]

Background: The emergence of drug-resistant bacteria has led to the search for newer plant-derived antimicrobial agents with lesser side effects. Andrographis paniculata (Nilavembu), the chief ingredient nilavembu kudineer has been successfully used in the treatment of dengue fever outbreak in India. Hence, this study was proposed to examine the antibacterial activity of A. paniculata against multidrug-resistant isolates of Gram-positive cocci (vancomycin-resistant Enterococcus faecalis (VRE) and methicillin-resistant Staphylococcus aureus [MRSA]) and Gram-negative bacilli (extended-spectrum beta-lactamase [ESBL]-producing Escherichia coli and Pseudomonas aeruginosa). Standard strains, ATCC 25922 E. coli, ATCC 25923 S. aureus, ATCC 27853 P. aeruginosa, and ATCC 29212 E. faecalis were used as controls. Methods: Extracts such as methanol, acetone, chloroform, and ethanol of A. paniculata (leaf) were prepared by cold percolation method. Agar well diffusion method was employed to assess the antibacterial activity. Minimum inhibitory concentration was determined using microbroth dilution method. Results: Results showed methanol extract exhibited maximum antibacterial activity against clinical and control strains. Phytochemical analysis of the methanol extract has confirmed the presence of an array of bioactive compounds. Conclusion: Methanolic extract of A. paniculata could serve as a potential herbal alternative in the management of infections caused by VRE and MRSA, while the ethanolic extract of A. paniculata could be used in the effective therapeutic management of ESBL-positive P. aeruginosa. [ABSTRACT FROM AUTHOR]

Background The threat posed by antibiotic resistance is of increasing concern in low- and middle-income countries (LMICs) as their rates of antibiotic use increase. However, an understanding of the burden of resistance is lacking in LMICs, particularly for multidrug-resistant (MDR) pathogens. Methods We conducted a retrospective, 10-hospital study of the relationship between MDR pathogens and mortality in India. Patient-level antimicrobial susceptibility test (AST) results for Enterococcus spp. Escherichia coli , Staphylococcus aureus , Klebsiella pneumoniae , Acinetobacter baumannii , Pseudomonas aeruginosa , and Enterobacter spp. were analyzed for their association with patient mortality outcomes. Results We analyzed data on 5103 AST results from 10 hospitals. The overall mortality rate of patients was 13.1% (n = 581), and there was a significant relationship between MDR and mortality. Infections with MDR and extensively drug resistant (XDR) E. coli , XDR K. pneumoniae , and MDR A. baumannii were associated with 2–3 times higher mortality. Mortality due to methicillin-resistant S. aureus (MRSA) was significantly higher than susceptible strains when the MRSA isolate was resistant to aminoglycosides. Conclusions This is one of the largest studies undertaken in an LMIC to measure the burden of antibiotic resistance. We found that MDR bacterial infections pose a significant risk to patients. While consistent with prior studies, the variations in drug resistance and associated mortality outcomes by pathogen are different from those observed in high-income countries and provide a baseline for studies in other LMICs. Future research should aim to elucidate the burden of resistance and the differential transmission mechanisms that drive this public health crisis. [ABSTRACT FROM AUTHOR]

Bile salt hydrolase (BSH) activity, hypo-cholesterolemic effect, and probiotic properties have been reported for Enterococcus strains isolated from animal and human gut and fermented foods but not for strains isolated from environmental niches, like aquatic and terrestrial plants, soil, and water. The present study is the first report on isolation of Enterococcus faecium from rhizospheric soils that harbor the bsh gene, remove cholesterol in vitro , and possess essential and desirable probiotic attributes. Fifteen samples were collected from different sites located in northern, southern, and central regions of India, of which five yielded pure colonies that were named LR2, LR3, ER5, LR13, and VB1. These were identified by 16S rRNA gene sequencing as E. faecium and evaluated for BSH activity, cholesterol-lowering potential in vitro , and probiotic properties. Our results indicated that all the strains were capable of surviving the harsh conditions of the gastrointestinal tract and did not harbor any of the virulence genes. Though all strains showed the presence of bsh and potential for cholesterol removal, E. faecium strain LR13 showed a remarkable cholesterol removal capability and vancomycin susceptibility and possessed most of the desirable and essential attributes of a probiotic. Hence, it seems to be a fairly promising probiotic candidate that needs to be further evaluated in in vivo studies, especially for its hypo-cholesterolemic potential. [ABSTRACT FROM AUTHOR]

At our hospital, a tertiary care hospital in north India, methicillin-resistant coagulase-negative staphylococci (CoNS) and vancomycin-resistant enterococci have become a worrisome clinical problem[[2]],[[3]]. Emergence of linezolid resistance in the vancomycin-resistant Enterococcus faecium multilocus sequence typing C1 epidemic lineage. [Extracted from the article]

Background: Enterococci, formerly classified with fecal streptococci, have been recognized to be of fecal origin since the beginning of this century. Method: This study was undertaken to determine the prevalence of stool colonization with vancomycin resistant Enterococcus (VRE) and also to evaluate the risk factors for colonization with vancomycin resistant Enterococcus among non-Hospitalized individuals at MMIMSR, Mullana. Test was performed for VRE isolates collected over a period of 6 months (Oct2015-March 2016). Faecal samples were collected by using sterile container from non-hospitalized individuals then to Cultures using Mac Conkey and Blood agar. After presumptive diagnosis as an enterococcus spp, 50 Enterococcal isolates were then again cultured on special VRE screen agar media to identify vancomycin resistant Enterococcus. Result: The results were further supported by modified Kirby-bauer disk diffusion method with vancomycin (30µg) as per CLSI guideline. A total of 29 (58%) Enterococcus faecalis and 21 (42%) Enterococcus faecium were detected among the faecal isolates and 2 (4%) were VRE. According to CLSI guideline isolates showing diameter of zone of inhibition =16mm were considered among the VRE. Chronic diseases, previous hospital stay (more than 15 days) and repeatedly antibiotic consumption was found to be significant risk factor for non-hospitalized individuals. Conclusion: There is need for programs to promote greater attention about antibiotics usage in the general population. Education of Health care workers with implementation and observation of hand-washing practices constitutes a very effective step in preventing the spread Prolonged use of vancomycin drug should not be recommended by the physician. [ABSTRACT FROM AUTHOR]

Introduction: Tigecycline Evaluation and Surveillance Trail (TEST) study is an on-going global surveillance. The study was performed to determine the susceptibility of common pathogens to tigecycline and comparator antibiotics by broth microdilution (BMD) at two tertiary care centres in India from 2015 to 2017. Methodology: Total of 989 isolates collected from various clinical specimens between January 2015 and September 2017 from two centres in India were included. BMD was performed to determine the minimum inhibitory concentration (MIC) for tigecycline and comparator antibiotics. Results: Among Gram-negative bacteria, susceptibility to tigecycline was lowest among Klebsiella spp. being 84% while others such as E. coli, Enterobacter spp., Serratia spp. and H. influenzae showed susceptibility of 98%, 95%, 98% and 100% respectively. Overall, 99 isolates among Enterobacteriaceae (E. coli, Klebsiella spp. and Enterobacter spp.) were ESBL producers, susceptible to tigecycline. Among the 101 meropenem resistant Enterobacteriaceae, 85 were susceptible to tigecycline (84%). Among the Gram-positive bacteria, S. aureus and Enterococcus spp. were 99% and 98% susceptible to tigecycline respectively. Among 68 MRSA isolates in the study, 66 (97%) were susceptible to tigecycline. Seven vancomycin resistant E. faecalis were isolated and all were susceptible to tigecycline. Conclusion: Tigecycline has retained activity over both Gram-positive and Gram-negative organisms with MIC values comparable to global reports. About 98% of the MDR Gram-positive and Gram-negative bacteria in the study are susceptible to tigecycline. With increased incidence of extensively drug resistant organisms, tigecycline is a potential reserve drug. [ABSTRACT FROM AUTHOR]

Introduction: Drug-resistant Enterococcus species is a persisting clinical problem and may serve as a reservoir of resistant genes. The present study was undertaken in Mangalore, India to know the antibiogram and genetic relatedness of Enterococcus spp. isolated from clinical samples. Methodology: A total of 150 non-repetitive Enterococcus spp. isolated from clinical samples were subjected to antimicrobial susceptibility testing by Kirby Bauer disk diffusion method. Molecular typing of the isolates was done by Random amplification of polymorphic DNA (RAPD). Results: Among the 150 isolates, 79 were from urine, 68 from pus and three from blood samples. Of this 58.7 % were E. faecalis and the remaining were E. faecium. Urinary isolates of E. faecium showed a higher percentage of antibiotic resistance when compared to E. faecium isolates from pus (p < 0.001). E. faecium from blood samples were resistant to ampicillin, penicillin, ciprofloxacin and were sensitive to vancomycin and teicoplanin. E. faecalis blood isolates were resistant to ciprofloxacin, penicillin, and erythromycin. 73% of Enterococcus isolates from pus were resistant to erythromycin. All the Enterococcus spp. were sensitive to vancomycin. Among the total Enterococcus isolates 44 were high-level aminoglycoside resistant (HLAR) by disc diffusion method which corresponded to MIC of > 500 μg/mL for gentamicin and > 1000 μg/mL for streptomycin. These isolates were subjected to RAPD, which showed similarity and differences in the banding patterns. Conclusions: Our study showed a baseline resistance among Enterococcus spp. in our area, which poses a challenge to the treating physicians and a reservoir for transmission of antibiotic resistant genes. [ABSTRACT FROM AUTHOR]

Background: A prospective observational study was carried out over a period of 6 months among the inpatients and outpatients in the departments of general medicine, urology, obstetrics, and gynecology of a tertiary level referral hospital in south Malabar region of Kerala to assess the prevalence of various bacterial isolates and their changing susceptibility to antimicrobial agents and to evaluate the pattern of antibiotic utilization in urinary tract infection (UTI). Methods: A total of 680 samples were analyzed from December 2015 to May 2016; among them, only 74 patients met the inclusion criteria. Of the 74 patients, 53 were female and 21 were male. Female patients were found to be significantly higher in number in the age group of 61–70years. The maximum number of UTI cases was reported from the general medicine department. Results: The disease was more significantly portrayed in suburbans (47%) and rural (38%) population compared with urbans (15%). Fever (60.81%) and dysuria (48.64%) were the most common symptoms. Diabetes (34%) was found to be the major predisposing factor followed by postmenopausal factors (24%), previous UTI (14%), renal disease (9%), benign prostatic hypertrophy (8%), smoking (7%), and oral contraceptives (4%). Escherichia coli was the major isolate (n=36) followed by Klebsiella (n=13), Citrobacter (n=8), Enterococcus (n=7), Enterobacter (n=4), Pseudomonas (n=3), coagulase-negative Staphylococcus (n=2), and Proteus (n=1). Majority of patients were prescribed with cefoperazone+sulbactum (n=28) followed by levofloxacin (n=25) and nitrofurantoin (n=10). Cumulative antibiogram was prepared as the final step of the study using the WHONET 5.6 software. Gram-negative organisms such as E. coli, Klebsiella, and Citrobacter showed higher susceptibility toward piperacillin-tazobactum, cefoperazone-sulbactam, imipenem, meropenem, amikacin, and netilmicin. Gram-positive organisms such as Enterococcus showed higher susceptibility toward piperacillin-tazobactum, cefoperazone-sulbactam, amoxicillin-clavulanic acid, vancomycin, and linezolid. Conclusions: In our setting, routine culture may be necessary as treatment failure with empirical therapy is likely to occur. [ABSTRACT FROM AUTHOR]

Purpose: To report Enterococcus faecalis endophthalmitis (EFE) in children and its comparison with adults.Methods: Retrospective study of 19 children up to 18 years age, undergoing vitrectomy for EFE at a referral eye center from January 1995 to December 2015. The results were compared to 18 adults with EFE.Results: The mean age was 7.4 ± 3.06 years. Post-traumatic EFE was seen in 17 (89.4%) children (p < 0.05) versus postoperative EFE seen in 12 (66%) adults. Broomstick 9 (52.9%) and hypodermic needle injury 6 (35.2%) were common associations. "Optimum functional outcome" was achieved in 12 (63.1%) eyes of children and 5 (27.7%) eyes of adults (p = 0.03). Sealed and Zone II wounds, lens, and needle injuries had good outcome whereas intervention after 3 days and retinal detachment had poor outcomes (p < 0.05). The was no correlation with the Ocular Trauma Score.Conclusion: EFE in children is rare; seen with broom stick and needle injuries, if managed early recovers optimum vision. [ABSTRACT FROM AUTHOR]

This study reports the polyphasic identification, characterization of virulence potential, and antibiotic susceptibility of Aeromonas salmonicida subspecies salmonicida COFCAU_AS, isolated from an aquaculture system in India. The physiological, biochemical, 16s rRNA gene sequencing and PAAS PCR test identified the strain as Aeromonas salmonicida. The MIY PCR tests established the subspecies as ' salmonicida '. The in vitro tests showed the isolated bacterium as haemolytic with casein, lipid, starch, and gelatin hydrolysis activity, indicating its pathogenic attributes. It also showed the ability to produce slime and biofilm, and additionally, it possessed an A-layer surface protein. In vivo pathogenicity test was performed to determine the LD 50 dose of the bacterium in Labeo rohita fingerlings (14.42 ± 1.01 g), which was found to be 106.9 cells fish−1. The bacteria-challenged fingerlings showed skin lesions, erythema at the base of the fins, dropsy, and ulcer. Almost identical clinical signs and mortalities were observed when the same LD 50 dose was injected into other Indian major carp species, L. catla and Cirrhinus mrigala. Out of the twelve virulent genes screened, the presence of nine genes viz., aerA , act , ast , alt , hlyA , vapA, exsA , fstA , and lip were detected , whereas ascV, ascC , and ela genes were absent. The A. salmonicida subsp. salmonicida COFCAU_AS was resistant to antibiotics such as penicillin G, rifampicin, ampicillin, and vancomycin while highly sensitive to amoxiclav, nalidixic acid, chloramphenicol, ciprofloxacin, and tetracycline. In summary, we have isolated a virulent A. salmonicida subsp. salmonicida from a tropical aquaculture pond which can cause significant mortality and morbidity in Indian major carp species. • Isolation of A. salmonicida subsp. salmonicida strain COFCAU_AS form India. • In vitro and in vivo tests confirmed the virulent nature of COFCAU_AS. • COFCAU_AS possess 9 tested virulent genes: aerA , act , ast , alt , hlyA , vapA, exsA , fstA lip. • COFCAU_AS was resistant to penicillin G, rifampicin, ampicillin, and vancomycin. [ABSTRACT FROM AUTHOR]

Introduction: Urinary tract infection (UTI) is one of the most common infectious diseases. With the emergence of multidrug resistance (MDR), therapeutic options for treatment of UTIs are becoming limited. Fosfomycin has emerged as a novel oral therapeutic option with bactericidal activity against the MDR uropathogens. We evaluated the susceptibility pattern of uropathogens to this antibiotic. Methods: A prospective study was conducted for 6 months in a tertiary care hospital in Eastern India to evaluate whether the common uropathogens were susceptible to fosfomycin. Identification of organisms causing significant bacteriuria was done by conventional biochemical and VITEK 2 Compact System™. Antimicrobial susceptibility testing was performed against these pathogens by Kirby-Bauer disc diffusion method. Minimum inhibitory concentrations were measured for certain drugs by E-strips and VITEK 2 Compact System. Results: A total of 2229 urine samples were referred for culture during the study period, which yielded 356 significant bacterial isolates. Among these isolates, 64.78% were extended-spectrum beta-lactamases producers, 15.97% were carbapenem-resistant Enterobacteriaceae, and 42.7% isolates were found to be MDR Enterobacteriaceae (MDRE). However, 95.18% of the total isolates and 95.93% of MDRE were found to be susceptible to fosfomycin. Conclusion: The common uropathogens, including MDR isolates, show high in vitro susceptibility to fosfomycin, which therefore has the potential to emerge as a promising alternative oral agent for outpatient therapy of UTIs. [ABSTRACT FROM AUTHOR]

Tunneled catheters are a lifeline in ESRD patients undergoing hemodialysis. Intravascular access devices have significant potential for producing iatrogenic diseases resulting in catheter related blood stream infections (CRBSI). CRBSI, a nosocomial infection is a significant clinical problem which is continuously evolving. They are associated with significant morbidity, mortality, and cost. The risk factors for blood stream infections in tunneled cuffed catheters include the duration of the catheter, past catheter related bacteraemia, hypoalbuminemia, immunosuppression. Gram positive organisms are the most common causative pathogens. Staph aureus CRBSI is more likely to require hospitalization or ICU admission. Metastatic infection like infective endocarditis, osteomyelitis, septic arthritis, spinal epidural abscess is relatively uncommon and most reported cases are caused by Staphylococcus aureus. Treatment of complicated CRBSI with sepsis, persistently positive blood cultures, or metastatic infection requires a longer course of antibiotics (6–8 weeks) compared with uncomplicated CRBSI. A 67 year old patient on dialysis via a tunneled catheter for 3 months duration presented with complaints of severe lower back pain. Blood cultures from catheter site and peripheral site grew Enterococcus fecalis. MRI spine was suggestive of epidural abscess. The tunneled catheter was removed and antibiotics were given based on the sensitivity reports. The differential diagnoses were multiple myeloma, TB spine, secondaries spine, osteoporosis. MRI contrast clinched the diagnosis of spinal epidural abscess, which is very uncommonly caused by Enterococcus fecalis. Atypical presentation of CRBSI should be kept in mind in all patients on intravascular access devices. Appropriate treatment with antibiotics and removal of the catheter is to be done if metastatic infection is present. [ABSTRACT FROM AUTHOR]

The article offers information on the immune dysregulation among Human immunodeficiency virus (HIV) patients that results in increased risk of morbidity due to S. aureus that causing bacterial bloodstream infections (BSIs). It mentions about the third‑generation cephalosporins and carbapenems as treatment options, which drives the emergence of multidrug‑resistant (MDR) bacteria.

Background: Herbs have attracted attention as a potential natural health-care approach that focuses on protecting and restoring the health. Caesalpinia bonducella is an Indian herb reported in Ayurveda, the ancient medicine system of India. This plant have extensive uses in folk medicines for the treatment of a variety of diseases such as pneumonia and filarial infections. Objective: This study was aimed to assess the antimicrobial activity of ethanolic extract of C. bonducella seed in the oral pathogens of human. Materials and Methods: C. bonducella leaf ethanolic extract was used to identify the antibacterial activity. Then, the C. bonducella extract was dissolved in dimethyl sulfoxide to get 1 mg/ml concentration. After 50-200 ml volume of the C. bonducella leaf extract was loaded on to a sterile disc containing human oral pathogens respectively. Agar well diffusion method was used to determine the minimized inhibitory concentration. The antibacterial activity of C. bonducella was studied. After 24 h incubation, the plate was observed and the zone was measured for the calculation of minimum inhibitory concentration of the drug. The data were compared with respective pathogens. Result: In this study, C. bonducella leaf ethanolic extract dose-dependently increased the bacterial growth inhibition. However, 100% inhibition was observed against Enterococcus faecalis than the standard drug ciprofloxacin. Finally, the extract showed a potential antibacterial activity. Conclusion: The use of herbal medicine is increasing with an increase in the development of drug resistance among bacterial population. Herbal medicines are safe with no side effects and have significant action against bacteria and other microorganisms. Based on the results recorded in the present findings, it is concluded that C. bonducella has a potential antimicrobial agent on human oral pathogenic microorganisms, and hence, the herbal drug may serve as one of the potential antimicrobial agents. [ABSTRACT FROM AUTHOR]

The goal of this study was to determine the unique characteristics of Enterococcus faecium MC-5, a probiotic bacteria isolated from the intestine of a fish, Cyprinus carpio specularis , collected from Dal Lake in Srinagar, Kashmir, India. For this, the important valuable probiotic attributes, some functional properties, and safety assessments were analyzed in-vitro for the strain MC-5. The strain E. faecium MC-5 exhibited high resistance to low pH, high bile salt, lysozyme, and phenol. The strain MC-5 showed excellent auto- and co-aggregation properties and displayed remarkable hydrophobicity towards various tested hydrocarbons which suggested that the strain possesses venerable adhesion properties. Apart from these, the cell-free supernatant (CFS) of strain MC-5 exhibited phenomenal antimicrobial activity against the tested pathogens. A scanning electron microscope (SEM) image revealed strain MC-5 finely adhered to human colon adenocarcinoma cells (HCT-15 cells). The strain MC-5 showed high bile salt hydrolase activity and excellent cholesterol removal ability of 70.27%. The intact cells of strain MC-5 also showed strong DPPH scavenging activity. The EPS produced by E. faecium MC-5 inhibited the adhesion of Listeria monocytogenes, Staphylococcus aureus, and Salmonella enterica on HCT-15 cells with maximum inhibition rates of 41.82, 40.34, and 55.51%, respectively for displacement assay, which was higher as compared to exclusion (26.06, 26.11, and 39.23%) and competition assays (30.06, 26.7, and 41.20%). Strain MC-5 did not exhibit hemolysis and was also found susceptible to vancomycin and other clinically important antibiotics. When evaluating all the results from the present study, it is propounded that strain MC-5 has enviable probiotic characteristics and thus can be used as bio-protective cultures and/or bio-shield in food and pharmaceutical industries. • Enterococcus faecium MC-5 was isolated from the intestine of a fish, Cyprinus carpio specularis. • The strain E. faecium MC-5 exhibited high resistance to low pH, high bile salt, lysozyme, and phenol. • SEM image revealed that strain MC-5 finely adhered to human colon adenocarcinoma cells (HCT-15 cells). • The strain MC-5 showed high bile salt hydrolase activity and excellent cholesterol removal ability of 70.27%. • Strain MC-5 did not exhibit hemolysis and was also found susceptible to vancomycin and other relevant antibiotics. [ABSTRACT FROM AUTHOR]

Background: Infection or colonization with multidrug-resistant organisms (MDRO) is associated with high mortality and morbidity. Knowledge of MDRO colonization may help in planning empirical antibiotic approach in neutropenic patients, which is known to improve patient outcomes. While routine cultures are positive and may help direct antibiotic therapy in only up to 15% neutropenic patients, surveillance cultures are positive in more than 90% of cancer patients.Aims: To assess the rate of MDRO carrier status at presentation and rate of conversion to MDRO during the treatment.Materials and Methods: Rectal swabs of all the outpatients presenting to pediatric oncology unit were sent within 7 days from date of registration from January 2014 to December 2014. Furthermore, stool cultures/rectal swabs of all patients who got directly admitted to the pediatric ward at presentation were sent within 24 h. Repeat rectal swabs were sent again for patients from this cohort when they got readmitted to the ward at least 15 days after last discharge or when clinically indicated.Results: Baseline surveillance rectal swabs were sent for 618 patients, which included 528 children with hematological malignancies and 90 children with solid tumors. Forty-five (7.3%) showed no growth. Of the remaining 573, 197 (34.4%) patients were colonized by two organisms and 30 (5.2%) by three organisms. Three hundred and thirty-four (58.4%) showed extended spectrum beta-lactamase (ESBL) Enterobacteriaceae, of which 165 (49.5%) were ESBL sensitive to beta-lactam with beta-lactamase inhibitors combinations and 169 (50.5%) were resistant to combinations. One hundred and sixteen (20.2%) were carbapenem-resistant Enterobacteriaceae (CRE) and 65 (11.4%) had vancomycin-resistant enterococci in baseline cultures. Only 63 (21%) patients were colonized by a sensitive organism in their baseline surveillance cultures. Morbidity (Intensive Care Unit stay) and mortality was higher in patients colonized by MDR organisms. There was a significant correlation between the place of residence and CRE colonization status with the highest rate (60%) of CRE colonization observed in children from East India. The repeat cultures showed the further conversion of sensitive isolates to MDRO in 80% of these children, of which 40% each converted from non-ESBL and non-CRE to ESBL and CRE, respectively.Conclusion: This is the first study illustrating the alarming high prevalence of community-acquired MDRO colonization, especially CRE, which has grave implications for therapy for children with cancer potentially compromising delivery of aggressive chemotherapy and affecting outcomes. This incidence further increases during the course of treatment. Knowing the baseline colonization also guides us for the planning of chemotherapy as well as antibiotic approach and infection control strategies. Local antibiotics stewardship including education of the healthcare workers as well as national level interventions to prevent antibiotic misuse in the community is critical to minimize this problem. [ABSTRACT FROM AUTHOR]

Abstract: We assessed the occurrence of pollution indicators and antibiotic resistant bacterial isolates from water and sediment samples of three different eco-regions of the Chennai coast between March – May of 2010. Total of 960 bacterial strains belonging to four genera were isolated which show the highest frequencies of resistance to vancomycin (53.6%) and penicillin (52.6%) (except Enterococcus sp., which is highly resistant to erythromycin) and lowest frequencies of resistance to chloramphenicol (3.43%), ciprofloxacin (3.95%), gentamicin (4.68%), and tetracycline (6.97%). The E. coli, Vibrio sp., Salmonella sp. and Enterococcus sp. show high frequency of resistance to 2–5 antibacterials of 60.4%, 45.83%, 69.16% and 46.6%, respectively. High pollution indices (PI – 6.66–14.06) and antibiotic resistance indices (ARI – 0.29–0.343) indicate that the coastal environment is highly exposed to antibiotic sources that suggesting to avoid direct contact. [Copyright &y& Elsevier]

In line with developments in the water reuse sector, this paper applied quantitative microbial risk assessment (QMRA) techniques to seven greywater reuse systems used for recycling shower water for toilet flushing. The objective of the study was to establish a scientific basis for health-based greywater targets for India. It involved qualitative risk assessments and quantitative microbiological analysis using Enterococci, thermotolerant coliforms (TTC) and coliphage indicator organisms. Four conclusions are drawn from this study. Firstly, the systems indicated a low risk and high quality, and secondly that low levels of risk were present in systems, resulting in the recommendation of guideline of 5000 cfu/100 mL rather than 10 000 cfu/100 mL for greywater utilised for direct toilet flushing. Thirdly, disability adjusted life years (DALYs) are a useful indicator of risk for evaluating the performance of a greywater reuse system in addition to chemical/microbiological indicators DALYs, and finally that TTC are a useful surrogate microbial indicator for analysis of greywater in developing countries with limited analytical facilities. [ABSTRACT FROM AUTHOR]