Vancomycin and Linezolid-Resistant Enterococcus Isolates from a Tertiary Care Center in India.

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.

Microbiological Profile in Patients Having Keratitis in a Tertiary Care Hospital in India.

Background Corneal ulcer or keratitis is defined as a loss of corneal epithelium with underlying stromal infiltration and suppuration associated with signs of inflammation. Corneal blindness is a significant public health problem worldwide; infectious keratitis is one of the predominant preventable causes of blindness. Several studies have evaluated microbial infectious keratitis's etiology, management, and outcome. However, there are regional variations in corneal ulcers' prevalence, risk factors, and outcome. The objective of this study was to isolate and identify the bacterial, fungal, viral, and protozoal etiological organisms causing infectious corneal ulcers along with their prevalence and antimicrobial sensitivity pattern. Methods A prospective observational study was done in the Department of Microbiology and RIO, Medical College & Hospital, Kolkata, for a period of 1 year (February 2019 to January 2020) after obtaining clearance from the Institutional Ethics Committee. Informed consent, demographic data, history of disease onset, duration of symptoms, associated co-morbidities, etc., were taken from the patients fulfilling the inclusion criteria. Corneal scraping samples were collected sterilely to detect bacterial, fungal, parasitic, and viral isolates and identified by standard laboratory procedures. Results A total of 80 patients were included in the study. The risk factors included foreign body in 24 (30%), blunt trauma in 10 (12.5%), steroid use in 8 (10%), contact lens user 4 (5%), and spontaneous in 34 (42.5%). Among these 80 patients, 18 showed growth of bacteria, including Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, and Pseudomonas aeruginosa; four had growth of fungi, including Aspergillus spp. and Fusarium spp, and two were positive for Herpes simplex virus by IFA. Conclusion Early diagnosis and prompt keratitis treatment are critical for preventing visual loss. The identification of the various causative agents of keratitis is essential for the proper management of the cases.

Bacteriologic Profile Along With Antimicrobial Susceptibility Pattern of Pediatric Pyoderma in Eastern India.

Background Pyogenic skin infection (pyoderma) is a bacterial infection of the skin and its appendages. Primary pyoderma is caused by the direct invasion of healthy skin, whereas secondary pyoderma originates in diseased skin as superimposed conditions, such as scabies, pediculosis, wounds, insect bites, and eczema. This study aimed to identify the clinical patterns and risk factors of pyoderma in a pediatric population and to isolate various causative bacteria and determine their susceptibility patterns. Methodology A prospective study was performed at the Medical College and Hospital, Kolkata, India, for one year (from August 2016 to July 2017), which included all children younger than 12 years with pyoderma attending the outpatient dermatology department (as the study was conducted among the pediatric population, only children below 12 years of age were included). Sterile cotton swabs were used to aseptically collect exudates or pus from lesions and anterior nares, which were used for culture, identification, and antibiotic susceptibility testing of the causative organisms. Results During the study period, a total of 182 patients were included, 121 (66.48%) of whom had primary pyoderma and 61 (33.52%) of whom had secondary pyoderma. Of the 182 patients, 161 showed bacterial growth on culture: 126 (78.26%) were Staphylococcus aureus, 18 (11.18%) were coagulase-negative staphylococci, 16 (9.94%) were Streptococcus pyogenes, and 1 (0.62%) was Pseudomonas aeruginosa. All staphylococci were susceptible to vancomycin and linezolid. Conclusions The most common cause of pyoderma in the pediatric age group is S. aureus, although the prevalence of methicillin-resistant S. aureus was low in this hospital. Proper identification and antibiogram are required for managing these cases.

Isolation, characterization and acyclovir susceptibility of herpes simplex virus isolates among immunocompromised patients.

INTRODUCTION: Herpes simplex virus (HSV) Type 2 primarily causes genital herpes, while HSV Type 1 is responsible for oral and facial lesions. The objective of this study was to isolate and characterize HSV from herpetic lesions among human immunodeficiency virus (HIV) infected patients and to evaluate their acyclovir susceptibility pattern. MATERIALS AND METHODS: Blister fluid and swabs from ulcers were collected from patients with clinical diagnosis of HSV infection among patients attending the HIV clinic of two tertiary care centers - Medical College, Kolkata, and School of Tropical Medicine, Kolkata. These samples were cultured in the Vero cell line. Growth of virus was noted by observing the characteristic cytopathic effect of HSV, which was further confirmed by immunofluorescence and polymerase chain reaction (PCR). These isolates were then subjected to the Vero cells with serial dilutions of acyclovir for determining the susceptibility pattern. RESULTS: Among the 52 samples received, 8 (15.38%) showed growth of HSV. After confirmation by immunofluorescence and PCR, all seven isolates from genital samples were identified as HSV-2 and the lone isolate from oral lesion was confirmed as HSV 1. Out of the eight isolates, 25% showed resistance to acyclovir. The overall isolation rate was more from genital blister than genital ulcer which was 46.15% and 2.86%, respectively. CONCLUSION: HSV was isolated in 15.38% of cases of clinical herpes. There was a higher isolation rate of virus from blister fluid as compared to ulcer scrapings. Acyclovir resistance in 25% of cases is alarmingly high.

Biofilm Producing Enterococcus Isolates from Vaginal Microbiota.

BACKGROUND: Enterococcus is an important cause of infection in the hospital as well as in the community. METHODS: A prospective study was done in Medical College, Kolkata for a period of 2 years (from January 2018 to December 2019). After obtaining clearance from the Institutional Ethics Committee, Enterococcus isolates from cases of vaginitis were included in the study. Identification of Enterococcus species was done by Gram stain and conventional biochemical tests along with automated identification by VITEK 2 Compact. These isolates were tested for antimicrobial susceptibility to different antibiotics by Kirby Bauer disc diffusion method and minimum inhibitory concentration (MIC) by VITEK 2 Compact. Interpretation of susceptibility was done according to the Clinical and Laboratory Standards Institute (CLSI) 2017 guidelines. Biofilm detection for Enterococcus species was done. RESULTS: During the period of 2 years, 39 isolates of Enterococcus spp. were obtained from vaginitis cases. Among these, 27 were Enterococcus faecalis and 12 Enterococcus faecium. All isolates were highly susceptible to vancomycin, teicoplanin, and linezolid. Biofilm was detected in eight isolates of which five were strong biofilm producer and three moderate biofilm producers. CONCLUSION: Biofilm production is an important virulence factor in Enterococcus isolates from vaginitis.

Diagnostic accuracy of commercially available immunochromatographic rapid tests for diagnosis of dengue in India.

BACKGROUND & OBJECTIVES: There is limited evidence regarding the accuracy of dengue rapid diagnostic kits despite their extensive use in India. We evaluated the performance of four immunochromatographic Rapid Diagnostic Test (RDTs) kits: Multisure dengue Ab/Ag rapid test (MP biomedicals; MP), Dengucheck combo (Zephyr Biomedicals; ZB), SD bioline dengue duo (Alere; SD) and Dengue day 1 test (J Mitra; JM). METHODS: This is a laboratory-based diagnostic evaluation study. Rapid tests results were compared to reference non-structural (NS1) antigen or immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) results of 241 dengue-positive samples and 247 dengue-negative samples. Sensitivity and specificity of NS1 and IgM components of each RDT were calculated separately and in combination (either NS1 or IgM positive) against reference standard ELISA. RESULTS: A total of 238, 226, 208, and 146 reference NS1 ELISA samples were tested with MP, ZB, SD, and JM tests, respectively. In comparison to the NS1 ELISA reference tests, the NS1 component of MP, ZB, SD, and JM RDTs demonstrated a sensitivity of 71.8%, 85.1%, 77.2% and 80.9% respectively and specificity of 90.1%, 92.8%, 96.1 %, and 93.6%, respectively. In comparison to the IgM ELISA reference test, the IgM component of RDTs showed a sensitivity of 40.0%, 50.3%, 47.3% and 20.0% respectively and specificity of 92.4%, 88.6%, 96.5%, and 92.2% respectively. Combining NS1 antigen and IgM antibody results led to sensitivities of 87.5%, 82.9%, 93.8% and 91.7% respectively, and specificities of 75.3%, 73.9%, 76.5%, and 80.0% respectively. INTERPRETATION & CONCLUSION: Though specificities were acceptable, the sensitivities of each test were markedly lower than manufacturers' claims. These results also support the added value of combined antigen-and antibody-based RDTs for the diagnosis of acute dengue.

Seroprevalence and Co-infection of Hepatitis B and Hepatitis C among Patients in a Tertiary Care Hospital in Eastern India.

INTRODUCTION: Hepatitis B (HBV) and Hepatitis C (HCV) are two common viral infections causing cirrhosis. AIM: The aim of this study was to find the seroprevalence of HBV and HCV along with occurrence of co-infection of HBV and HCV in patients attending a tertiary care hospital. MATERIALS AND METHODS: The study was done for a period of one year (January to December 2016) in the Department of Microbiology, Medical College, Kolkata. After obtaining ethical clearance and informed consent from the patients, serum samples were collected from all patients referred to Department of Microbiology for antibody to HCV and Hepatitis B surface antigen (HBsAg) screening. ELISA was performed for anti HCV antibody and HBsAg. The results and relevant clinical information were noted and analysis was done. RESULTS: A total of 10802 samples were received, of which 316 (2.92 %) were HBsAg positive, 115 (1.06%) were HCV antibody positive and a total of 7 (0.07%) patients were positive both for HBsAg and Anti HCV antibody. There was male preponderance. Anti HCV antibody was more common in age below 10 years and in thalassemia patients. Out of 7 patients positive for both, 5 patients were on regular blood transfusion due to beta thalassemia and 2 patients had history of chronic liver disease. CONCLUSION: In this study, it was found that there was seroprevalence of 2.92 % of HBsAg, 1.06% of HCV antibody and 0.07% positive both for HBsAg and HCV antibody among the patients of a tertiary care centre in Eastern India.

A randomized, double-blind trial of amorolfine 0.25% cream and sertaconazole 2% cream in limited dermatophytosis.

BACKGROUND: Dermatophytosis is becoming increasingly unresponsive to conventional antifungals. Newer topical antifungals may be more effective in these patients. AIMS: To evaluate and compare the efficacy and safety of amorolfine 0.25% cream and sertaconazole 2% cream in limited tinea cruris/corporis. METHODS: A single-center, randomized (1:1), double-blind, parallel group, active-controlled trial (CTRI/2014/12/005246) was performed. Sixty-six untreated adults with acutely symptomatic tinea cruris/corporis were included in the study. All patients had limited cutaneous involvement and were KOH mount positive. Group A received amorolfine 0.25% cream, and group B received sertaconazole 2% cream twice daily application to the lesions for 4 weeks. After the baseline visit, four follow-up visits were carried out. The outcome measures for effectiveness were clinical and mycological cure. Safety parameters studied were treatment-emergent adverse events and changes in routine laboratory parameters. RESULTS: Both sertaconazole and amorolfine significantly reduced symptoms (P < 0.001) in both groups. However, improvement in symptoms (pruritus, burning sensation, erythema, scaling and crusting) was significantly greater in the sertaconazole group at every follow-up visit. Sertaconazole cream was also more effective than amorolfine cream in reducing the number of lesions (P = 0.002 at 12 weeks) and improving the Dermatology Life Quality Index (P < 0.001) at all the follow-up visits. Adverse events were similar in the two groups (P = 0.117). Fungal cultures became negative in 92.3% of the sertaconazole group as compared to 80% in the amorolfine group (P = 0.010). LIMITATIONS: Antifungal susceptibility testing could not be done. CONCLUSION: Sertaconazole 2% is superior to amorolfine 0.25%, both in terms of effectiveness and tolerability. Improvement can be appreciated from second week onwards.

Facile synthesis of highly disperse Ni-Co nanoparticles over mesoporous silica for enhanced methane dry reforming.

A synergistic approach was made to develop a highly stable and carbon resistant catalyst system based on cobalt and nickel supported over modified mesoporous silica for the dry reforming of methane (DRM). Modified mesoporous silica is prepared by a hydrothermal method, and the total Co & Ni composition is taken at around 5% by using the deposition-precipitation technique. CO2 reforming with methane was performed at 400-800 °C under atmospheric pressure as well as at a pressure of 1 MPa, keeping the CH4/CO2 ratio equal to unity. The catalyst assembly before and after the reaction was thoroughly characterized by a wide range of analytical techniques including N2 physisorption, XRD, TPR, TPO, TPH, XPS, SEM, TEM, elemental mapping, TG-DTG. The physicochemical characterization results confirmed the homogeneous distribution of nanosized metal particles into the hexagonal framework of modified silica, which plays a vital role towards a stronger metal support interaction that renders carbon deposition upon the active metal surface as well as avoids metal sintering at higher temperatures. At the same time, the coexistence of nanosized Co and Ni into the mesopores produced a synergy which provides better stability without any deactivation at high pressure reaction conditions. In situ DRIFT analysis evidenced that the reaction proceeds over these catalysts through an initial pathway in which both methane and carbon dioxide initially dissociate over the metal along with a bifunctional pathway in which methane dissociates over the active metal and carbon dioxide activated over the basic support surface via a formate intermediate. Density Functional Theory (DFT) calculations were also performed and further support the proposed mechanism from DRIFT studies.

Cystic echinococcosis: A neglected disease at usual and unusual locations.

Echinococcus granulosus causes a zoonotic infection called cystic echinococcosis (CE) or more commonly known as hydatid disease. Although the two most common locations of hydatid cyst are liver and lung, it may also appear in other parts of the body. Clinical presentation of the hydatid disease depends on the site and size of the lesion. A retrospective study was done in Medical College and Hospital, Kolkata, from January 2012 to June 2014, to find the site of involvement, distribution, clinical features, history of contact, mode of presentation, laboratory diagnosis, and treatment modalities of the cases of hydatid cyst. The cases were identified by radiological and laboratory methods, the data were entered in Excel spreadsheet, and analysis was done. Among the 21 cases of hydatid cyst included in the study, solitary hepatic involvement was seen in 11 (52.38%), pulmonary involvement in 4 (19%), and 6 (28.71%) were in unusual locations such as liver cyst extending as retroperitoneal, omental cyst, choledochal cyst, splenic cyst, and in hepatorenal pouch. History of contact with dog was seen in 15 (71.43%). All the patients were treated with surgery and albendazole and were discharged in healthy condition. CE may be present in usual and unusual locations with a lot of variations in the clinical features. Hence, proper radiological and laboratory diagnosis is required for accurate diagnosis and appropriate management of these cases.

Can EDTA Change MRSA into MSSA? A Future Prospective!

INTRODUCTION: In the present era we are left behind with limited options for the treatment of serious infections caused by multidrug resistant S.aureus, most remarkably nosocomially acquired Methicillin resistant S.aureus (MRSA). The problem increases more when these strains easily become multidrug resistant (MDR) due to biofilm formation. Those staphylococcal species that are vancomycin and linezolid resistant are also resistant to other antistaphylococcal agents which call for an urgent intervention to develop newer antimicrobial agents. AIM: The present study was undertaken with the aim to evaluate the antibiofilm effect of EDTA against the biofilm forming MRSA isolates, isolated from different clinical infections. MATERIALS AND METHODS: The biofilms formed on polystyrene microtitre plates by the MRSA strains were treated by different concentrations of EDTA to find out its anti-biofilm activity. Further simultaneously the antibiotic susceptibility pattern was noted down to check whether the MRSA strains become MSSA (Methicillin sensitive S.aureus). RESULTS: Our data demonstrates that EDTA at 4mM concentration inhibits biofilm of MRSA and at 20 mM have an ability to reduce and dissociate the biofilm membrane, allowing the antibiotics to enter and convert MRSA strains into MSSA. CONCLUSION: These findings suggest that commercially available EDTA could be used in future to control MRSA and its biofilm- related infections.

Seroprevalence of Herpes Simplex Virus Infection in HIV Coinfected Individuals in Eastern India with Risk Factor Analysis.

Herpes simplex virus type 2 (HSV-2) is the cause of most genital herpes while HSV-1 is responsible for orolabial and facial lesions. In immunocompromised individuals, like HIV patients, impaired immunity leads to more frequent symptomatic and asymptomatic HSV infection. Fifty-two blood samples from HIV patients with clinically diagnosed HSV infection were taken as cases, while 45 blood samples each from HIV-infected (HIV control) and noninfected patients without any herpetic lesion (non-HIV control) were taken as control. Serum was tested for IgM and IgG antibodies of both HSV-1 and HSV-2 by ELISA. The seroprevalence was compared among the three groups of study population, considering the demographic and socioeconomic parameters. The HSV-2 IgM was significantly higher (p < 0.005) in the HIV patient group (34.6%) than the HIV control (2.2%) and non-HIV control (2.2%) groups, whereas HSV-2 IgG seroprevalence was higher in both HIV patient (61.5%) and HIV control (57.8%) groups than the non-HIV control group (17.8%). The prevalence of HSV-2 was significantly higher in persons with multiple partners and in the reproductive age group. The overall seroprevalence of HSV-1 IgM was too low (<5%), whereas it was too high (about 90%) with HSV-1 IgG in all three study groups.

In vitro activity of levofloxacin against lower respiratory tract pathogens.

BACKGROUND: Considerable morbidity and mortality are associated with lower respiratory tract infections (LRTIs) that put a considerable strain on the health budget. Selection of appropriate antibiotics as empirical therapy maximizes positive patient outcomes, and that depends on regular surveillance of infective agents and their antibiograms, which vary according to the geographical areas. AIM: The aim was to study the drug susceptibility pattern of the isolated pathogens of the respiratory tract infections. SETTINGS AND DESIGN: Retrospective study for a period of 1-year 3 months from January 2013 to March 2014 at a Tertiary Care Hospital. MATERIALS AND METHODS: Eleven hundred and eighty-four sputum samples from both outdoor and indoor patients with symptoms of LRTI were processed, and antibiotic sensitivity test was done to commonly used antibiotics. Descriptive statistics was used to analyze the data. RESULTS: Among 502 quality sputum samples, 312 (62.15%) samples showed growth of pathogenic bacteria. The most common pathogens were Klebsiella spp. (38.14%), Moraxella spp. (16.02%), Streptococcus pneumoniae (14.10%), Pseudomonas spp. (9.93%), S. aureus (9.29%). It was found that the overall susceptibility pattern was <50% for amoxicillin, amoxicillin-clavulanic acid, cefuroxime, cotrimoxazole and erythromycin whereas for cefotaxime, cefixime, and cefoperazone-sulbactum it was 60.08%, 51.59%, 69.04%, respectively. The susceptibility to ciprofloxacin, ofloxacin, and levofloxacin were 66.67%, 70.19% and 83.33%, respectively. CONCLUSION: Klebsiella spp. was the most common LRTI pathogen. There was limited activity of amoxicillin, amoxicillin-clavulanic acid, cefuroxime, cotrimoxazole and erythromycin for the treatment of LRTI whereas levofloxacin, (being an oral drug with good compliance) had good activity against respiratory pathogens and could be used for empiric treatment in LRTI.

Cutaneous Nocardia brasiliensis infection in an immunocompetent host after ovarian cystectomy: A case study.

Nocardia brasiliensis is a rare human pathogen that is usually associated with localised cutaneous infections. We report a case of primary cutaneous Nocardia brasiliensis infection causing delayed wound healing that developed after ovarian cystectomy in an otherwise healthy 32-year-old woman. The patient was initially treated with cotrimoxazole, however due to intolerance intravenous amikacin was given and gradually the wound healed. The diagnosis was confirmed by demonstrating the causative organism in exudates, and cultures. Early diagnosis as well as early institution of chemotherapy is effective in most patients, and antimicrobial susceptibility testing of the isolate should be performed to identify the best treatment options.