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Introduction: Healthcare personnel have the right to decent, healthy, and safe working conditions during COVID-19. Despite our best efforts to safeguard them against SARS-CoV-2 infection, a substantially large number of healthcare personnel fell ill and succumbed to COVID-19. This study aimed to characterize the SARS-CoV-2 exposures among healthcare personnel and determine the risk level of those events, thereby identifying and focusing on the key areas that influence workplace safety. Materials and Methods: A pre-tested questionnaire was used to collect data from healthcare personnel on the type of exposure, place, and use of preventive measures, along with demographic and occupational data. Exposure events were categorized into low, moderate, and high-risk by using a risk categorization protocol. Results: Doctors were primarily found to be involved in the aerosol-generating procedure, which had 94 times higher odds (95% CI: 43.9926–201.17) for high-risk exposures than other activities. In contrast, nurses were more likely to experience close-contact exposures (OR: 2.77, 95% CI: 1.44–5.33). Both critical care units and operation theaters were identified to have higher odds (OR: 2.34, 95% CI: 1.33–4.23 and OR: 2.31; 95% CI: 0.99–5.42, respectively) than the wards for high-risk exposures. Use of personal protective equipment was poor, with breaches reported in 10.2% of all exposure events. The lacunae in practice were addressed by repeated training and counseling and by ensuring the adequacy of resources in required areas. Conclusions: The assessment was beneficial for safeguarding healthcare personnel and assuring a safer workplace during the early phase of the COVID-19 pandemic.

IntroductionThe emergence of disinfectant resistance has become a severe threat due to reduced effectiveness. This study was undertaken to determine how bacteria adapt to survive exposure to disinfectants in the busiest section of a tertiary care hospital in Varanasi, India.MethodsFour isolates (two Klebsiella pneumoniae, Kp1 and Kp2; two Pseudomonas aeruginosa, Pa1 and Pa2) were obtained from chlorhexidine (CHX)–based handwash during microbiological surveillance of “in-use disinfectants” in hospital. Six disinfectants [4% CHX, 2% glutaraldehyde, 7.5% hydrogen peroxide, 1% sodium hypochlorite and 0.1% benzalkonium chloride (BAC), and 70% ethyl alcohol] were tested against these four isolates to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Antibiotic profile, change in MIC on exposure to disinfectants and biofilm formation in the presence and absence of disinfectants was studied. Whole genome sequencing (WGS) was done to identify the resistance mechanisms.ResultThe isolates showed the highest MBC/MIC ratio (4) against glutaraldehyde. Exposure to supra-inhibitory concentration of BAC for 21 days resulted in doubling of MIC/MBC. The majority (75%) of the isolates were multidrug resistant. All the isolates were strong biofilm producers. The reduction rate of biofilm formation decreased with an increase in the concentration of disinfectants (p = 0.05 for BAC). WGS revealed multiple AMR genes including blaDIM-1, disinfectant-resistant gene and efflux pump genes.ConclusionThe study emphasized the various adaptation strategies of these isolates for survival in disinfectant environment, thus posing a huge challenge for their control in the hospital environment.

Abstract Infections by Entamoeba histolytica (E. histolytica) lead to considerable morbidity and mortality worldwide and treatment is reliant on a single class of drugs, nitroimidazoles. Treatment failures and intermittent reports of relapse from different parts of world indicate towards development of clinical drug resistance. In the present study, susceptibility testing of clinical isolates of E. histolytica was carried against metronidazole and tinidazole. Additionally, anti-amoebic property of active compounds of Andrographis paniculata was also evaluated. Prevalence of metronidazole resistance gene (nim) in patients attending hospital was also done to get comprehensive insight of present situation of drug resistance in E. histolytica. Mean inhibitory concentration 50 (IC50) value of E. histolytica isolates against metronidazole and tinidazole was 20.01 and 16.1 µM respectively. Andrographolide showed minimum mean IC50 value (3.06 µM). Significant percentage inhibition of E. histolytica isolates by andrographolide was seen as compared to metronidazole (p = 0.0495). None of E. histolytica isolates showed presence of nim gene. However, in stool samples from hospital attending population, prevalence of nimE gene was found to be 76.6% (69/90) and 62.2% (56/90) in diarrheal and non-diarrheal samples respectively. Inhibitory concentration of commonly used nitroimidazoles against clinical isolates of E. histolytica are on rise. Percentage inhibition of E. histolytica isolates by andrographolide was significantly higher than control drug metronidazole.

Abstract The bubbling community of microorganisms, consisting of diverse colonies encased in a self-produced protective matrix and playing an essential role in the persistence of infection and antimicrobial resistance, is often referred to as a biofilm. Although apparently indolent, the biofilm involves not only inanimate surfaces but also living tissue, making it truly ubiquitous. The mechanism of biofilm formation, its growth, and the development of resistance are ever-intriguing subjects and are yet to be completely deciphered. Although an abundance of studies in recent years has focused on the various ways to create potential anti-biofilm and antimicrobial therapeutics, a dearth of a clear standard of clinical practice remains, and therefore, there is essentially a need for translating laboratory research to novel bedside anti-biofilm strategies that can provide a better clinical outcome. Of significance, biofilm is responsible for faulty wound healing and wound chronicity. The experimental studies report the prevalence of biofilm in chronic wounds anywhere between 20 and 100%, which makes it a topic of significant concern in wound healing. The ongoing scientific endeavor to comprehensively understand the mechanism of biofilm interaction with wounds and generate standardized anti-biofilm measures which are reproducible in the clinical setting is the challenge of the hour. In this context of “more needs to be done”, we aim to explore various effective and clinically meaningful methods currently available for biofilm management and how these tools can be translated into safe clinical practice.

Objective Coagulase-negative staphylococci (CoNS) are being implicated as one of the leading causes of bloodstream infection (BSI). To study the spectrum, prevalence, and antimicrobial susceptibility of CoNS causing BSI in neonates.

Abstract Background The emergence of multidrug-resistant tuberculosis (MDR-TB) has complicated the situation due to the decline in potency of second-line anti-tubercular drugs. This limits the treatment option for extensively drug-resistant tuberculosis (XDR-TB). The aim of this study was to determine and compare the minimum inhibitory concentration (MIC) by agar dilution and resazurin microtiter assay (REMA) along with the detection of mutations against linezolid and clofazimine in confirmed XDR-TB clinical isolates. Results A total of 169 isolates were found positive for Mycobacterium tuberculosis complex (MTBC). The MIC was determined by agar dilution and REMA methods. The isolates which showed non-susceptibility were further subjected to mutation detection by targeting rplC gene (linezolid) and Rv0678 gene (clofazimine). The MIC for linezolid ranged from 0.125 µg/ml to > 2 µg/ml and for clofazimine from 0.25 µg/ml to > 4 µg/ml. The MIC50 and MIC90 for linezolid were 0.5 µg/ml and 1 µg/ml respectively while for clofazimine both were 1 µg/ml. The essential and categorical agreement for linezolid was 97.63% and 95.26% and for clofazimine, both were 100%. The sequencing result of the rplC gene revealed a point mutation at position 460 bp, where thymine (T) was substituted for cytosine (C) while seven mutations were noted between 46 to 220 bp in Rv0678 gene. Conclusion REMA method has been found to be more suitable in comparison to the agar dilution method due to lesser turnaround time. Mutations in rplC and Rv0678 genes were reasons for drug resistance against linezolid and clofazimine respectively.

Abstract Background Extensive drug resistance in Klebsiella pneumoniae (K. pneumoniae) causing major outbreaks in large hospitals is an emerging challenge. We describe a near fatal outbreak of colistin resistant, carbapenem resistant K. pneumoniae (CRKp) producing metallo beta-lactamases (bla NDM) and bla OXA-48 in the neonatal intensive care unit (NICU) at the background of a larger outbreak involving multiple parts of the hospital and the challenges in its containment. Methods Following identification of an outbreak due to colistin resistant CRKp between April to June 2017 in the NICU, a thorough surveillance of similar cases and the hospital environment was performed to trace the source. All the isolated K. pneumoniae were tested for susceptibility to standard antibiotics by disc diffusion and microbroth dilution methods. Molecular detection of extended spectrum beta lactamases (ESBLs) and carbapenemases (classes A, B, D) genes was done. Enterobacterial repetitive intergenic consensus (ERIC) PCR and multi-locus sequence typing (MLST) was done to determine the genetic relatedness of the isolates. Characteristics of different sequence types were statistically compared (Student’s t-test). Results A total of 45 K. pneumoniae isolates were studied from NICU (14 cases of neonatal sepsis), ICU (18 cases), other wards (7 cases) along with 6 isolates from hospital environment and human colonizers. The primary case was identified in the ICU. All the K. pneumoniae from NICU and 94.4% from the ICU were colistin resistant CRKp. Majority (59.37% and 56.25%) harbored bla SHV/bla CTXM and bla OXA-48 genes, respectively. Two distinct sequence types ST5235 and ST5313 were noted with colistin resistance, distribution within the NICU and mortality as significant attributes of ST5235 (p

BackgroundThe rapid emergence of carbapenem resistant Acinetobacter baumannii (CRAB) has resulted in an alarming situation worldwide. Realizing the dearth of literature on susceptibility of CRAB in genetic context in the developing region, this study was performed to determine the susceptibility profile against standard drugs/combinations and the association of in-vitro drug synergy with the prevalent molecular determinants.Methods and findingsA total of 356 clinical isolates of A. baumannii were studied. Confirmation of the isolates was done by amplifying recA and ITS region genes. Susceptibility against standard drugs was tested by Kirby Bauer disc diffusion. Minimum inhibitory concentration (MIC), MIC50 and MIC90 values against imipenem, meropenem, doripenem, ampicillin/sulbactam, minocycline, amikacin, polymyxin B, colistin and tigecycline was tested as per guidelines. Genes encoding enzymes classes A (blaGES, blaIMI/NMC-A, blaSME, blaKPC), B (blaIMP, blaVIM, blaNDM) and D (blaOXA-51,blaOXA-23 and blaOXA-58) were detected by multiplex polymerase chain reaction. Synergy against meropenem-sulbactam and meropenem-colistin combinations was done by checkerboard MIC method. Correlation of drug synergy and carbapenemase encoding genes was statistically analyzed.ResultsOf the total, resistance above 90% was noted against gentamicin, ciprofloxacin, levofloxacin, ceftazidime, cefepime, ceftriaxone, cotrimoxazole and piperacillin/tazobactam. By MIC, resistance rates from highest to lowest was seen against imipenem 89.04% (n=317), amikacin 80.33% (n=286), meropenem 79.49% (n=283), doripenem 77.80% (n=277), ampicillin/sulbactam 71.62% (n=255), tigecycline 55.61% (n=198), minocycline 14.04% (n=50), polymyxin B 10.11% (n=36), and colistin 2.52% (n=9). CRAB was 317 (89.04%), 81.46% (n=290) were multidrug resistant and 13.48% (n=48) were extensively drug resistant. All the CRAB isolates harboured blaOXA-51 gene (100%) and 94% (n=298) blaOXA-23 gene. The blaIMP gene was most prevalent 70.03% (n=222) followed by blaNDM, 59.62% (n=189). Majority (87.69%, 278) were co-producers of classes D and B carbapenemases, blaOXA-23 with blaIMP and blaNDM being the commonest. Synergy with meropenem-sulbactam and meropenem-colistin was 47% and 57% respectively. Reduced synergy (p=

ABSTRACT The rapid emergence of drug resistance in Acinetobacter baumannii has put forward the use of colistin as a last-resort treatment for infections with A. baumannii. Empirical colistin use without prior susceptibility testing has been one of the factors that has been promoting drug resistance in low-resource settings. In this regard, while the advocated broth microdilution (BMD) method for colistin susceptibility testing is often considered cumbersome, the preferable colistin broth disk elution (CBDE) method has not yet been approved for A. baumannii. To prevent the underreporting of colistin susceptibility, we tested the CBDE method for A. baumannii and compared the results with those of BMD. A total of 125 A. baumannii, including 100 susceptible and 25 resistant isolates were tested via the CBDE method and compared with the standard BMD method. The essential agreement, categorical agreement, sensitivity, and specificity for CBDE were 97.6% (n = 122), 98.4% (n = 123), 100%, and 98.40%, respectively. The percentage of major error found was 1.6% (n = 2), and no very major error was found. CBDE in A. baumannii could be considered in low-resource settings. IMPORTANCE The relatively cumbersome broth microdilution (BMD) method for routine colistin susceptibility testing has not been adopted, especially in low-resource settings, often leading to the underreporting of colistin susceptibility and the promotion of the empirical use of colistin. In this regard, the much-preferred colistin broth disk elution (CBDE) method has not yet been approved for A. baumannii. We evaluated colistin susceptibility via the CBDE method, compared the results with those of the BMD method in 125 A. baumannii isolates with various profiles, and inferred that the CBDE method using 50 μL inoculum could be helpful, at least in resource-limited setups, versus not reporting susceptibility testing for colistin.

Currently, the world is facing a challenge to curb the outbreak of the highly contagious COVID-19, which has significantly impacted many lives and is affecting the practice of healthcare professionals as well. During this containment phase, an influx of COVID-19 patients requiring surgical interventions is expected. To combat the situation, every health care staff must implement and adhere to various infection prevention and control measures. In this article, we have addressed the importance of basic infection prevention and control practices required for handling COVID-19 cases, especially, those requiring acute surgical care. On one hand, certain non-emergency procedures need to be delayed, on the other hand, all surgeries during this period need to be proactively performed with appropriate infection control precautions regardless of the COVID-19 status of the patient. Prudent decisions and a conservative approach while making the best choices regarding surgeries will not only ensure personal protection but will also preserve important resources, manpower, hospital beds, and equipment for repurposing during this urgent crisis.

Infection prevention and control (IPC) program is obligatory for delivering quality services in any healthcare setup. Lack of administrative support and resource-constraints (under-staffing, inadequate funds) were primary barriers to successful implementation of IPC practices in majority of the hospitals in the developing countries. The Coronavirus Disease 2019 (COVID-19) brought a unique opportunity to improve the IPC program in these hospitals. A PDSA (Plan-Do-Study- Act) model was adopted for this study in a tertiary care hospital which was converted into a dedicated COVID-19 treatment facility in Varanasi, India. The initial focus was to identify the deficiencies in existing IPC practices and perceive the opportunities for improvement. Repeated IPC training (induction and reinforce) was conducted for the healthcare personnel (HCP) and practices were monitored by direct observation and closed-circuit television. Cleaning audits were performed by visual inspection, review of the checklists and qualitative assessment of the viewpoints of the HCP was carried out by the feedbacks received at the end of the training sessions. A total of 2552 HCP and 548 medical students were trained in IPC through multiple offline/onsite sessions over a period of 15 months during the ongoing pandemic. Although the overall compliance to surface disinfection and cleaning increased from 50% to >80% with repeated training, compliance decreased whenever newly recruited HCP were posted. Fear psychosis in the pandemic was the greatest facilitator for adopting the IPC practices. Continuous wearing of personal protective equipment for long duration, dissatisfaction with the duty rosters as well as continuous posting in high-risk areas were the major obstacles to the implementation of IPC norms. Recognising the role of an infection control team, repeated training, monitoring and improvisation of the existing resources are keys for successful implementation of IPC practices in hospitals during the COVID-19 pandemic.

Frequent outbreaks of food-borne pathogens, particularly E. coli O157:H7, continue to impact human health and the agricultural economy tremendously. The required cell count for this pathogenic strain of E. coli O157:H7 is relatively low and hence it is vital to detect at low colony forming unit (CFU) counts. Available detection methods, though sensitive, fall short in terms of timeliness and often require extensive sample processing. To overcome these limitations, we propose a novel magneto-plasmonic nanosensor (MPnS) by integrating surface plasmon resonance (SPR) properties with spin–spin magnetic relaxation (T2 MR) technology. We engineered MPnS by encapsulating several gold nanoparticles (GNPs) within the polymer-coating of iron oxide nanoparticles (IONPs). First, the polyacrylic acid (PAA)-coated IONPs were synthesized using a solvent precipitation method, then gold chloride solution was used to synthesize GNPs and encapsulate them within the PAA-coatings of IONPs in one step. A magnetic separation technique was used to purify the MPnS and the presence of GNPs within IONPs was characterized using transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDS), and other spectroscopic methods. The synthesized MPnS exhibits MR relaxation properties while possessing amplified optical properties than conventional GNPs. This allows for rapid and ultrasensitive detection of E. coli O157:H7 by SPR, T2 MR, and colorimetric readout. Experiments conducted in simple buffer and in milk as a complex media demonstrated that our MPnS-based assay could detect as low as 10 CFUs of this pathogenic strain of E. coli O157:H7 in minutes with no cross-reactivity. Overall, the formulated MPnS is robust and holds great potential for the ultrasensitive detection of E. coli O157:H7 in a simple and timely fashion. Moreover, this platform is highly customizable and can be used for the detection of other foodborne pathogens.

BackgroundEntamoeba infections have major impact on millions of the people worldwide. Entamoeba histolytica has long been accepted as the only pathogenic species. However, recent reports of other Entamoeba spp. in symptomatic cases have raised questions on their pathogenicity.Methodology/principal findingsTotal 474 stool samples and 125 liver aspirates from patients with intestinal and extra intestinal manifestations and from community were included. Sewage samples from the hospital and the city were also included. Microscopic examination and molecular detection were performed to detect presence of E. histolytica/ dispar/ moshkovskii/ bangladeshi. The associated demographic and socioeconomic factors were statistically analyzed with the presence of Entamoeba. Microscopy detected Entamoeba spp. in 5.4% stool and 6.4% liver aspirate samples. Through nested multiplex PCR, prevalence of Entamoeba spp. in intestinal and extra-intestinal cases was 6.6% (20/301) and 86.4% (108/125) respectively and in asymptomatic population was 10.5% (13/123). Sewage samples did not show presence of any Entamoeba spp. Uneducated subjects, low economic conditions, untreated drinking water, consumption of raw vegetables and habit of not washing hands before meals were significantly associated with presence of Entamoeba spp.ConclusionsE. histolytica still remains the only Entamoeba spp. in invasive extra intestinal infections. E. dispar was detected in both asymptomatic and symptomatic intestinal infections. Routine identification of Entamoeba spp. should incorporate PCR based detection methods.

The recent emergence of multidrug-resistant (MDR) Klebsiella pneumoniae with hypervirulent traits causing severe infections and considerable mortality is a global cause for concern. The challenges posed by these hypermucoviscous strains of K. pneumoniae with regard to their optimal treatment, management, and control policies are yet to be answered. We studied a series of extensively drug-resistant (XDR) and hypervirulent K. pneumoniae ST5235 isolates with resistance to carbapenems and polymyxins causing neonatal sepsis in a tertiary care hospital in India. A total of 9 K. pneumoniae isolates from 9 cases of neonatal sepsis were studied with respect to their clinical relevance, antimicrobial susceptibility profile, presence of extended spectrum β lactamase (ESBL) production, and responsible genes, carbapenemases (classes A, B, and D), and aminoglycoside-resistant genes. Hypervirulence genes encoding hypermucoid nature, iron uptake, and siderophores were detected by multiplex PCR. The plasmid profile was studied by replicon typing. Isolates were typed by multilocus sequence typing (MLST) and enterobacterial repetitive intergenic consensus (ERIC) PCR to study the sequence types (STs) and clonal relation, respectively. The neonates in the studied cases had history of pre-maturity or low birth weight with maternal complications. All the cases were empirically treated with piperacillin–tazobactam and amikacin followed by imipenem/meropenem and vancomycin and polymyxin B as a last resort. However, all the neonates finally succumbed to the condition (100%). The studied isolates were XDR including resistance to polymyxins harboring multiple ESBL genes and carbapenemase genes (blaNDM and blaOXA−48). Hypervirulence genes were present in various combinations with rmpA/A2 genes present in all the isolates. IncFI plasmids were detected in these isolates. All belonged to ST5235. In ERIC PCR, 6 different clusters were seen. The study highlighted the emergence and burden of XDR hypervirulent isolates of K. pneumoniae causing neonatal sepsis in a tertiary care hospital.

Drug resistance in tuberculosis (TB) is the biggest global health challenge as it hinders the tuberculosis control program and makes the disease more worsen. Molecular methods interrupt the spread of drug resistance by facilitating the appropriate anti- tuberculosis therapy at correct time through rapid diagnosis of multi drug resistant (MDR) and extensively drug resistant tuberculosis (XDR-TB). In this study we standardized and evaluated the diagnostic utility of multiplex allele specific PCR (MAS-PCR) targeting gyrA D94G and rrs A1401G mutations for detection of resistance against two key drugs (ofloxacin and kanamycin) of second line anti tuberculosis treatment. MAS-PCR assays targeting gyrA D94G and rrs A1401G for ofloxacin (OFL) and kanamycin (KAN) resistance respectively were carried out on 150 multidrug resistant isolates of Mycobacterium tuberculosis. The results were compared with phenotypic drug susceptibility test against ofloxacin and kanamycin by using proportion method on MGIT 960. Of 150 MDR isolates 50 were resistant to both ofloxacin and kanamycin, 36 were resistant to ofloxacin only, 8 were resistant to kanamycin only and 56 were susceptible to both the drugs. MAS-PCR correctly identified gyrA D94G and rrs A1401G mutations in phenotypically resistant isolates with a specificity of 100%. The sensitivity of MAS-PCR was 88.66%, 93.55% and 86% for OFL, KAN and XDR-TB respectively. There was no mutation detected at gyrA D94G region of 12.86% (11 of 86) OFL resistant isolates while 6.89% (4 of 58) of KAN resistant isolates did not carry rrs A1401G substitution. MAS-PCR proves to be a rapid tool for detection of drug resistance which could also be used as an initial marker for screening of XDR-TB. Keywords: Drug susceptibility test, Extensively drug-resistant tuberculosis, Ofloxacin, Kanamycin

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We report an unusual case of adult Loa loa infection in the anterior chamber of the eye of a 65-year-old female from rural area of Jharkhand, India, with no history of travel to endemic places but the presence of microfilaria of Loa loa in blood. Loiasis is endemic to African countries, with few sporadic cases affecting the Indian population. However, in none of the reports, microfilaremia was detected. This atypical feature in this case adds to the epidemiological variations in parasitic presentations in nonendemic regions.

Abstract The aim of this research is twofold: 1) to shed light on zika’s binding and entry mechanism while 2) demonstrating the effectiveness of our magnetic relaxation platform to achieve this goal. Magnetic relaxation-sensitive nanoparticles (MRNPs) are used in a novel fashion to analyze binding interactions between the zika envelope protein (ZENV) and proposed host cell receptors: AXL, HSP70, and TIM-1. Computational analysis is also utilized to examine these binding interactions for the first time. In addition, the role of crizotinib as a potential binding inhibitor is demonstrated and the possibility of ligand-independent phosphatidylserine-mediated binding is explored. Our findings suggest that while the extracellular domain of AXL has the highest affinity for ZENV; HSP70, TIM-1, and phosphatidylserine might also play active roles in zika tropism, which offers a potential explanation for the variety of zika-associated symptoms. This is, to our knowledge, the first time that MRNPs have been used to examine and quantify host-zika interactions. Our magnetic relaxation platform allows for timely and sensitive analysis of these intricate binding relationships, and it is easily customizable for further examination of additional host-pathogen interactions.

BackgroundAmebiasis, caused by Entamoeba histolytica (E. histolytica), is a significant cause of morbidity and mortality in developing countries. Mortality due to amebiasis is mostly by extra intestinal infections, amebic liver abscess being the most common one. This study was conducted to determine the current epidemiological status, risk factors, associated microflora and strain variation of E. histolytica causing liver abscesses.Methods/findingsA total of 115 liver abscess cases comprising of 107 (93%) males and 8 (6.9%) females were included in the study. Microscopic examination of pus samples from the abscesses and species discrimination using nested multiplex PCR showed the presence of E. histolytica in 101 (87.5%) cases. Data collected by face to face interviews using a pre tested questionnaire suggested intake of untreated drinking water (ORs: 6.4, p = 0.002), habit of alcohol consumption (ORs: 4.0, p = 0.019) and lack of urban services (ORs: 0.08, p = 0.017) to be major risk factors associated with E. histolytica infections. The study of associated bacterial flora through aerobic culture of liver aspirates and conventional PCR for detection of anaerobes revealed the presence of Fusobacterium (19, 25.5%), Peptococcus (19, 25.5%), Prevotella (18, 24.3%), Bacteroides (8, 10.8%), Staphylococcus aureus (3, 4%), Escherichia coli (2, 2.7%), Peptostreptococcus (2, 2.7%), Clostridium (2, 2.7%) and Klebsiella pneumoniae (1, 1.3%). Further to study the clonality, genotyping of E. histolytica targeting six tRNA-linked polymorphic STR loci (A-L, D-A, N-K, R-R, STGA -D and S-Q) was carried out which showed the presence of 89 different genotypes in the liver aspirate samples.ConclusionThe findings highlight the high prevalence of genetically diverse E. histolytica from the liver abscess cases in this geographical region. Low socio-economic status and habit of alcohol consumption were important predictors of amebic liver abscess.

The increasing emergence of Acinetobacter spp. with healthcare associated infections (HCAI) in intensive care units (ICU) is alarming. This study was a laboratory-based audit to determine the prevalence of Acinetobacter spp. associated with HCAI in the adult ICU of a tertiary care hospital in Varanasi, north India, with special reference to antimicrobial resistance and resistance determinants over a period of 5 years. A total of 993 cases of HCAI were analyzed. Isolates were characterized as multidrug resistance and extended drug resistance (MDR/XDR) based on antimicrobial susceptibility records. Few (100) randomly selected isolates of Acinetobacter baumannii (A. baumannii) were tested for imipenem, meropenem, and polymyxin B susceptibility by minimum inhibitory concentration (MIC) and for the presence of class A and B carbapenemases by multiplex PCR. Active surveillance of ICU environment was also performed. High prevalence of Acinetobacter related hospital acquired pneumonia (HAP) with significant resistance to imipenem (p

Background: Alarming rise of vancomycin-resistant enterococci (VRE) is a global cause of concern. Several factors have been held responsible for such rise, of which antibiotic usage is a prominent one. Objectives: This study was undertaken to determine the intestinal VRE colonization rate amongst hospitalized patients in relation to use of various antibiotics in the Intensive Care Unit (ICU) of a tertiary care university hospital, India. Materials and Methods: Stool samples were collected weekly from all the patients in the adult ICU for a period of 6 months and processed for isolation and phenotypic and genotypic characterization of VRE isolates. Patient and treatment details were noted and cases (those with VRE in stool) and controls (those without VRE in stool) were compared statistically. Further, a multivariate analysis was done to identify those antibiotics as independent risk factors for VRE colonization. Results: VRE colonization was found in 34.56% (28/81) of the patients studied, with the majority 75% (21/28) carrying the vanA gene. The cases had significantly more (P < 0.05) duration of hospital stay and antibiotic exposure. Intake of metronidazole, vancomycin, and piperacillin-tazobactam were identified as significant risk factors both in univariate and multivariate analysis. Conclusion: A potential reservoir of VRE was thus revealed even in low VRE prevalence setting. Based on this high colonization status, restriction of empirical antibiotic use, reviewing of the ongoing antibiotic policy, and active VRE surveillance as an integral part of infection control strategy were suggested.

The increasing reports of vancomycin-resistant enterococci (VRE) as a cause of neonatal septicemia are of recent interest. However, in majority of the cases, the source of VRE could not be located. As a consequence, the real importance of VRE and its control measures is undermined. Herein, we report a case of neonatal septicemia due to VRE (Enterococcus faecalis) of vanA genotype with VRE carriage in stool of the neonates as a possible source of sepsis. The report put forwards some lacunae in the infection control practices that are presently followed in the country.

Introduction: The ubiquity and widespread distribution of Intestinal Parasitic Infections (IPIs) is equally alarming despite tremendous efforts for their control. Periodic surveillance is required to detect the change in trends of IPIs based on different age groups. Aim: To determine the prevalence of IPIs among patients in different age groups attending the various departments of a tertiary care hospital in Varanasi. Materials and Methods: This retrospective study was conducted in the Microbiology Department, Institute of Medical Sciences and associated University hospital over a period of 5 years. Non-repetitive, consecutive stool samples from 2723 patients that had been sent for parasitic examination were processed and examined by microscopy as per standard. Prevalence of IPI was studied in age groups 06-15 years, 16- 30 years, 31-50 years, 51-70 years and above 70 years and data was statistically analyzed using Chi-Square test (Medcalc software version 15.11.14). Results: The prevalence of IPIs among patients attending the hospital was 8.63% (235/2723 cases), of which the most frequent infection was due to Entamoeba spp (62, 26.38%) followed by hookworm (59, 25.11%). Prevalence of infections with Giardia (2.69%) and Hymenolepis nana (2.69%) was significantly more (p16 years). Conclusion: Widespread distribution of intestinal parasites is still a major problem. Along with soil transmitted helminths (STHs), giardiasis is a significant problem in children of school going age group. Besides mass drug administration for STHs in this age group, policies should also be directed to treat and eradicate Giardia infections.

Purpose: The purpose of this study was to determine the prevalence of various Candida species and study some of their virulence factors among thevulvovaginal candidiasis(VVC)patients. Study Design and Settings: The study was conducted in a Tertiary Care University Hospital in North India. Materials and Methods: This study was carried out prospectively for a period of 1 year. High vaginal swabs (HVSs) were collected from women in childbearing age group attending the gynecology and obstetrics out-patient departments with the complaints suggestive of vulvovaginitis. Samples were plated on Sabouraud′s dextrose agar slope. Candida spp. isolated was further speciated based on microscopy, biochemical tests and culture characteristics on special media. Virulence factors of these strains were determined by biofilm formation and phospholipase activity. Result: A total of 464 HVS from 232 patients with the complaints of vulvovaginitis were included in this study. Following laboratory workup, 71 specimens were positive for genus Candida (30.6%). Further speciation showed 32.4% as Candida albicans, 45.07% Candida parapsilosis and 22.53% of Candida glabrata. Biofilm production was shown by 50 candidal strains (70.4%) and phospholipase activity was given by 41 candidal strains (57.74%). Conclusion: Our study suggests increasing prevalence of non-albicans Candida among the VVC cases along with their virulence factors. Therefore, we recommend that microbiological investigation upto species level should be mandatory to determine the emergence of non-albicans Candida as a major cause of VVC.

Drug resistance in tuberculosis is a major public health challenge in developing countries. The limited data available on drug resistance in extra pulmonary tuberculosis stimulated us to design our study on anti-tuberculosis drug resistance pattern in cases of extra pulmonary tuberculosis in a tertiary referral hospital of North India. We performed Geno Type MTBDRplus assay in comparison with conventional drug susceptibility testing by proportion method to study the mutation patterns in rpoB, katG and inhA genes.A total of 510 extra pulmonary samples were included in this study. After the smear microscopy, all the specimens were subjected for culture on Lowenstein Jensen (LJ) media. Phenotypic drug susceptibility testing (DST) was performed on LJ media for all the MTB isolates and compared with the results of Geno Type MTBDRplus assay which was performed with the DNA isolated from the culture by conventional method.Of 510 specimens cultured, the total culture positivity obtained was 11.8% (60) encompassing 54 (10.6%) Mycobacterium tuberculosis and 6 (1.2%) non-tubercular mycobacteria (NTM). DST results by Geno Type MTBDRplus assay and solid culture methods were compared in 51 MTB isolates excluding the two Rif indeterminate and one invalid test. Geno Type MTBDRplus accurately identified 13 of 14 rifampicin-resistant strains, 14 of 15 isoniazid-resistant strains and 13 of 14 as multi drug resistant tuberculosis (MDR-TB) in comparison with conventional method. Sensitivity and specificity were 92.86% and 97.30% respectively for detection of RIF resistance, 93.33% and 94.44% respectively for detection of INH resistance, 92.86% and 97.30% respectively for detection of MDR-TB, while the overall concordance of Geno Type MTBDRplus assay with conventional DST was 94.11%. The turn-around time for performing Geno Type MTBDRplus assay test was 48 hours.The problem of MDR in extra pulmonary tuberculosis (EPTB) cannot be overlooked and due attention on patients should be given. Routine use of Geno Type MTBDRplus assay for the diagnosis of MDR-EPTB can substantially reduce the time between diagnosis and drug therapy. Culture along with Geno Type MTBDRplus assay could be a solution for rapid and accurate diagnosis of MDR-TB in low bacillary non sputum specimens.

Background: Enterococci, though they are a part of commensal flora, are becoming increasingly important as nosocomial pathogens, due to their inherited and acquired resistances to several antimicrobial agents. In this context, Enterococcus faecium (E.faecium) requires a special mention due to its characteristic of Multidrug Resistance (MDR) and its ability to disseminate. Aim: This study was undertaken to phenotypically characterize and determine clonal relatedness amongst the indoor isolates of Enterococcus faecium (E. faecium) which were isolated from patients with urinary tract infections (UTIs). Settings and design: This study was carried out prospectively in a tertiary care university hospital and in Department of Microbiology, Varanasi, India. Material and Methods: Urine samples were collected from patients who were admitted in different departments of the hospital with a clinical diagnosis of UTIs and they were processed for a period of one year. Enterococcal species were identified by doing extensive biochemical tests. Anti–microbial susceptibility testing was done by disc diffusion and agar dilution methods. Molecular typing of the isolates was done by Random Amplified Polymorphic DNA (RAPD) typing method. Results: A total of 48 Enterococcal urinary isolates were identified in indoor patients, among which a majority (46, 95.83%) were E.faecium isolates. These isolates exhibited high resistance to fluoroquinolones (91.3%) and to ampicillin (60.86%) in particular. Two isolates were found to be resistant to vancomycin on screen agar. RAPD typing showed two major clusters, one of which had ten strains of 100% similarity, all of which were isolated from a common source. Conclusion: This study showed dissemination of multidrug resistant E. faecium isolates within the hospital. Being a quick and cost effective method, RAPD typing can be used to show clonal relatedness and to trace possible sources of organisms for epidemiological purposes.

Along with emergence of multidrug resistance, presence of several virulence factors in enterococci is an emerging concept. This study was undertaken to determine the prevalence of various virulence factors phenotypically and genotypically in enterococci and study their association with multidrug resistance. A total of 310 enterococcal isolates were studied, comprising 155 E. faecium and 155 E. faecalis. Antimicrobial susceptibility testing was done by disc diffusion and agar dilution method. Hemolysin, gelatinase, biofilm production, and haemagglutination were detected phenotypically and presence of virulence genes, namely, asa1, gelE, cylA, esp, and hyl, was detected by multiplex PCR. Of the total, 47.41% isolates were high level gentamicin resistant (HLGRE) and 7.09% were vancomycin resistant (VRE). All the virulence traits studied were found in varying proportions, with majority in E. faecalis (p>0.05). Strong biofilm producers possessed either asa1 or gelE gene. gelE silent gene was detected in 41.37% (12/29). However, increase in resistance was associated with significant decrease in expression or acquisition of virulence genes. Further, acquisition of vancomycin resistance was the significant factor responsible for the loss of virulence traits. Though it is presumed that increased drug resistance correlates with increased virulence, acquisition of vancomycin resistance might be responsible for reduced expression of virulence traits to meet the “biological cost” relating to VRE.

A healthy young adult male presented with complaints of frequent (>3/day) formed stools and passage of excessive mucous in stool for 3 months. He did not complain of nocturnal motions, recent diarrhea, blood in stool, straining, weight loss, or pain abdomen. Stool test was normal. He was counseled and treated as a case of irritable bowel syndrome. Due to inadequate relief with empirical therapy, colonoscopy was performed in a subsequent visit. Club-shaped small, round organisms with moving proboscis were seen in the cecum. Organism was later identified as a trematode Gastrodiscoides hominis, a rare foodborne trematode. The patient was treated with praziquantel, without complete relief. Trematode infection might not be the cause of symptoms.

To generate a cytopathic effect, the catalytic A1 subunit of cholera toxin (CT) must be separated from the rest of the toxin. Protein disulfide isomerase (PDI) is thought to mediate CT disassembly by acting as a redox-driven chaperone that actively unfolds the CTA1 subunit. Here, we show that PDI itself unfolds upon contact with CTA1. The substrate-induced unfolding of PDI provides a novel molecular mechanism for holotoxin disassembly: we postulate the expanded hydrodynamic radius of unfolded PDI acts as a wedge to dislodge reduced CTA1 from its holotoxin. The oxidoreductase activity of PDI was not required for CT disassembly, but CTA1 displacement did not occur when PDI was locked in a folded conformation or when its substrate-induced unfolding was blocked due to the loss of chaperone function. Two other oxidoreductases (ERp57 and ERp72) did not unfold in the presence of CTA1 and did not displace reduced CTA1 from its holotoxin. Our data establish a new functional property of PDI that may be linked to its role as a chaperone that prevents protein aggregation.

Cholera toxin (CT) travels as an intact AB(5) protein toxin from the cell surface to the endoplasmic reticulum (ER) of an intoxicated cell. In the ER, the catalytic A1 subunit dissociates from the rest of the toxin. Translocation of CTA1 from the ER to the cytosol is then facilitated by the quality control mechanism of ER-associated degradation (ERAD). Thermal instability in the isolated CTA1 subunit generates an unfolded toxin conformation that acts as the trigger for ERAD-mediated translocation to the cytosol. In this work, we show by circular dichroism and fluorescence spectroscopy that exposure to 4-phenylbutyric acid (PBA) inhibited the thermal unfolding of CTA1. This, in turn, blocked the ER-to-cytosol export of CTA1 and productive intoxication of either cultured cells or rat ileal loops. In cell culture studies PBA did not affect CT trafficking to the ER, CTA1 dissociation from the holotoxin, or functioning of the ERAD system. PBA is currently used as a therapeutic agent to treat urea cycle disorders. Our data suggest PBA could also be used in a new application to prevent or possibly treat cholera.