The hospital antibiogram is a periodic summary of antimicrobial susceptibilities of local bacterial isolates submitted to the hospital's clinical microbiology laboratory. Antibiograms are often used by clinicians to assess local susceptibility rates, as an aid in selecting empiric antibiotic therapy, and in monitoring resistance trends over time within an institution. Antibiograms can also used to compare susceptibility rates across institutions and track resistance trends. Some hospitals have adequate support from the computer department to be able to extract data from their reporting module. The WHONET software can be freely downloaded and used for analysis. Consensus guidelines have been developed by the Clinical and Laboratory Standards Institute (CLSI) to standardise methods used in constructing antibiograms. These guidelines can be incorporated into the WHONET software for analysis. Only the first isolate from the patient is to be included in the analysis. The analysis should be done on the basis of patient location and specimen type. The percentage susceptibility of the most frequently isolated bacteria should be presented in the antibiogram, preferably in a tabular form. The antibiogram must be printed or put up in the intranet for easy access to all clinicians. Antibiotic policy is one of the mandatory requirements for accreditation, and making an antibiogram is the first step before framing the antibiotic policy. The future of antibiograms would be the incorporation of patient related data to make information more reliable and for predicting outbreaks.

HIV and tuberculosis co-infection interact in fundamentally important ways. This interaction is evident patho-physiologically, clinically and epidemiologically. There are several differences between HIV-infected and HIV-uninfected patients with tuberculosis (TB) that have practical diagnostic implications. TB is more likely to be disseminated in nature and more difficult to diagnose by conventional diagnostic procedures as immunosuppression progresses. As TB rates continue to increase in HIV-endemic regions, improved diagnostic techniques merit consideration as TB-control strategies. There is a need to develop more user friendly techniques, which can be adapted for use in the high-burden and low-income countries. This review focuses on the diagnostic challenges in HIV-TB co-infection with an update on the current techniques and future prospects in an era of HIV pandemic.

Aims: To determine the prevalent subtypes of HIV-1 in serodiscordant couples. Setting: Integrated Counselling and Testing Centre (ICTC), Department of Microbiology. Study Design: Prospective pilot study. Participants: Thirty HIV-1 serodiscordant couples. Inclusion Criteria: a) Documentation of HIV-1 infection in one partner and seronegative status in the other, current history of continued unprotected sexual activity within the partnership, demonstration that they have been in a partnership for at least 1 year and are not currently on highly active antiretroviral therapy HAART; b) willingness of both partners to provide written informed consent including consent to continued couple counselling for 3 months. Materials and Methods: HIV-1 subtyping was carried out by heteroduplex mobility analysis (HMA) by amplifying env region; and DNA sequencing by amplifying gag region. Results: HIV-1 env gene was amplified successfully in 10/30 samples; gag gene, in 25/30 samples; and both env and gag gene were amplified successfully in 5/30 samples. HIV-1 subtype C was detected from 21 samples; subtype B, from 7; and subtype A, from 2. Sample from 1 positive partner was detected as subtype C by env HMA and subtype B by gag sequencing. Conclusion: HIV-1 subtype C was found to be the predominant subtype of HIV-1 in serodiscordant couples attending our ICTC, followed by HIV-1 subtype B and HIV-1 subtype A, respectively. DNA sequencing was found to be the most reliable method for determining the subtypes of HIV-1.

Purpose: Integrated counselling and testing centres (ICTC) provide counselling and blood testing facilities for HIV diagnosis. Oral fluid tests provide an alternative for people whodo not want blood to be drawn. Also, it avoids the risk of occupational exposure. The goal of this study was to evaluate the utility of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting. Materials and Methods: A cross-sectional study was carried out after ethics committee approval in 250 adult ICTC clients. Blood was collected and tested from these clients for HIV diagnosis as per routine policy and the results were considered as the gold standard. Also, after another written informed consent, oral fluid was collected from the clients and tested for the presence of HIV antibodies. Twenty five clients who had and 25 clients who had not completed their secondary school education (Group A and Group B, respectively) were also asked to perform and interpret the test on their own and their findings and experiences were noted. Result: The sensitivity, specificity, PPV and NPV of the oral fluid antibody test were 100%, 98.51%, 94.11% and 100%, respectively. Seventy six percent of clients preferred oral fluid testing. Group B found it difficult to perform the test as compared to Group A and this difference was statistically significant (P ≤ 0.05). Conclusion: Oral fluid testing can be used as a screening test for HIV diagnosis; however, confirmation of reactive results by blood-based tests is a must.

Introduction: Re-emergence of Chikungunya is a major public health problem in the southern states of India. Objectives: This study was undertaken to investigate an outbreak of Chikungunya, in June-August 2008 using PCR and determine the prevalent genotypes of Chikungunya virus (CHIKV) associated with the outbreak. Materials and Methods: Samples of blood were collected (in heparinized vacutainer tubes) from suspected patients of CHIKV infection from both Government Taluk Hospital in Kerala and a tertiary care hospital in Chennai, Tamil Nadu. A one-step RT-PCR was carried out on a block thermo-cycler targeting the E2 gene that codes for the viral envelope protein. The amplicons were verified for 305 bp size by standard agarose gel electrophoresis. The PCR products were purified, sequenced, and compared with other CHIKV strains reported from different geographical regions. A phylogenetic tree was constructed using MEGA 4. Results: Altogether 118 samples were collected from patients who presented with sudden onset of fever and/or joint pain, myalgia, and headache. CHIKV infection was confirmed by RT-PCR in 14 patients and all these cases were from Kerala. The positivity correlated with the early stage of the disease as all these patients had fever of less than seven days duration. The study isolates have been allotted the GenBank accession nos. GQ272368-GQ272381. Phylogenetic analysis of recent CHIKV isolates by partial sequencing of E2 region shows that isolates are closely related to strains from neighboring states and the African type. Conclusion: RT-PCR is a useful technique for the early detection of CHIKV infection during outbreaks. Molecular characterization of the strains indicates that majority of the strains have originated from the Central/East African strains of CHIKV.

Purpose: Tuberculosis remains an important health problem all over the world, especially in resource poor settings like India. The Ziehl-Neelsen (ZN) staining of sputum smear is still the method of choice in the diagnosis of tuberculosis in spite of its low sensitivity and specificity. This paper evaluates comparison of two different polymerase chain reaction (PCR) assays with sputum smear findings to detect Mycobacterium tuberculosis. Materials and Methods: A total of 191 sputum samples were collected from 84 patients attending a tertiary care hospital, who were suspected of having pulmonary tuberculosis, were examined by PCR targeting two different genomic regions, namely, TRC ₄ by non-nested format and IS6110 insertion element by nested format in comparison to ZN staining of sputum smears. Results: Among the patients tested, 20.24% (Mid-p 95%CI: 31.5-52.4) were smear positive, 7.14% (Mid-p 95%CI: 2.94-14.26) were positive by TRC ₄ PCR and 41.67% (Mid-p 95%CI: 12.7-29.8) were positive by IS6110 nested PCR (nPCR). The median age of overall positive cases was 42 years. Among the nPCR positives, the median for age of rural and peri-urban community was 46 and 32 years, respectively. The kappa coefficient between smear findings and TRC4 PCR findings was 0.27 and an agreement of 0.83 was observed (Z = 2.99; one-tailed P = 0.001). TRC ₄ PCR picked two unique positives that were negative by smear and IS6110 nPCR. Conclusion: The non-nested TRC ₄ PCR showed inability for accurate detection of M. tuberculosis in sputum samples. The study concluded that the nPCR targeting IS6110 is superior and more sensitive than TRC ₄ PCR.

Background: Mycobacterial antigen MPB64 is a secretory protein specific for Mycobacterium tuberculosis complex. A lateral flow immunochromatographic assay (ICA) is a method used for the rapid differentiation of M. tuberculosis complex. Aim: We aimed to evaluate the performance of ICA in rapid differentiation of M. tuberculosis complex from 97 Mycobacterium species other than tuberculosis (MOTT), which are grown in Lφwenstein-Jensen and TK-selective (SLC) medium. Materials and Methods: The study was performed in our laboratory between January 2009 and January 2010. A total of 394 isolates consisting of reference strains of 34 M. tuberculosis from World Health Organization (WHO) collection, 97 different MOTT bacilli, 7 Mycobacterium bovis BCG substrains and total 256 clinical Mycobacterium isolates were tested by ICA, which is based on anti-MPB64 monoclonal antibodies. All the strains were inoculated onto a TK-SLC (selective) medium and Lφwenstein-Jensen medium. TK-SLC is a new rapid mycobacterial culture medium that indicates mycobacterial growth by colour change. Results: The growth of mycobacterial strains was observed in 10-12 days on TK-SLC medium. ICA test was performed in 15 minutes. All strains belonging to M. tuberculosis complex group were found positive and all MOTT species were found negative on ICA slides. The results were confirmed with nucleic acid amplification by polymerase chain reaction (PCR) using primers specific for M. tuberculosis complex. Conclusion: With the additive effect of growth on TK-SLC medium in 10-12 days, the mycobacterial antigen MPB64 is a very useful and specific tool in rapid differentiation of M. tuberculosis and MOTT grown in culture.

Objective: Group B streptococci (GBS) from Southern African areas have been less well characterized. Our objective was to study serotype and serovariant distribution of carrier GBS strains as part of a study of the epidemiology of GBS carriage in pregnant women from Zimbabwe. Materials and Methods: We studied GBS isolated from 121 healthy pregnant women living in Harare and surrounding areas, Zimbabwe. Capsular polysaccharide (CPS) testing for serotype determination and surface-anchored protein testing for serosubtype determination were done by gene-based serotyping (PCR), except for the proteins R3 and a novel protein called Z, which were detected by antibody-based methods. Results: Strains of the CPS types Ia (15.7%), Ib (11.6%), II (8.3%), III (38.8%), V (24.0%) and NT (1.7%) were detected along with the strain-variable proteins Cί (15.7% of isolates), Cα (19.8%), Alp1 (epsilon-22.3%), Alp3 (5.0%), R4/Rib (46.3%), R3 (27.3%), Z (27.3%), and SAR5 (28.9%), which encodes the R5 protein. Up to four of the protein genes could be possessed or the gene product expressed by one and the same isolate. A total of 32 serovariants were detected. The findings assessed by us as most important were the very low prevalence of the gene Alp3 (Alp3 - 4.9%), high prevalence of R4 (Rib - 46.2%), the proteins R3 (27.3%), Z (27.3%), and of SAR5 (R5 - 28.9%). The low prevalence of Alp3, notably in GBS type V strains, differed from findings with CPS type V GBS from non-African areas. Bacteria of the various CPS types showed distinct CPS/protein-marker associations. Conclusion: The results are of importance in relation to regional variations of GBS phenotypes and genotypes and thus, of importance in planning and research in the context of future vaccine formulations.

Purpose: The purpose of the current study was to evaluate two low-costing PCR assays for rapid detection of Group B Streptococcus (GBS) in comparison to a pigment-based culture method. Materials and Methods: One-hundred and fifty vaginal swabs were collected from pregnant women at 35-40 weeks of gestation. Vaginal swabs were inoculated in selective enrichment broth medium, and examined using Islam medium, cfb PCR and scpB PCR assays. The demographic data were analysed to identify independent predictors of GBS colonization (age and gravidity), with GBS status as the dependent variable. Results: There was a significant association of age and gravidity with GBS colonization. GBS was detected in 25.3% of isolates by Islam medium, in 30.6% by using the cfb PCR assay and in 30% by using the scpB PCR assay. Conclusion: older pregnant women (≥30 years) and multigravida (>3 pregnancies) are at higher risk of GBS colonization. Both scpB-gene and cfb-gene-based PCR methods are highly sensitive techniques (100% sensitivity) compared to culture method. However, the specificities of the scpB and cfb PCR assays were 93.75 and 92.85%, respectively.

Purpose: To determine the prevalence of Chlamydia trachomatis infection in a high-risk population by direct and indirect methods and to evaluate the diagnosis of secretory immunoglobulin A (sIgA). Patients and Methods: Urethral or endocervical specimens from 78 patients (48 females and 30 males) were examined by cell culture, direct fluorescence assay, PCR Cobas Amplicor (Roche Molecular Diagnostics), and sIgA was detected by the recombinant lipopolysaccharide (LPS)-enzyme-linked immunoassay (rELISA). Serum from each patient was also obtained and analysed for the presence of IgG and IgA antibody by in-house microimmunofluorescence (MIF) and by the rELISA method (Medac, Hamburg, Germany). Results: The overall C. trachomatis prevalence determined by direct methods was 28%. The detection of sIgA antibodies was significantly higher in the group of patients with a positive direct detection (50%) than in the group of negative direct detection (10.7%). The Chlamydia-specific IgA antibodies were detected by the rELISA in 40.9 and 53.6% of group I (positive direct detection) and group II patients (negative direct detection), respectively. The species-specific IgA antibodies were detected by the MIF method in 18.2 and 16.1% of group I and II patients, respectively. Chlamydia genus-specific IgG antibodies were detected by the rELISA in 86.4 and 83.9% of group I and group II patients and, C. trachomatis specific IgG were present in 81.8 and 73.2% of group I and group II patients, respectively, as assessed by the MIF test. Conclusion: Combining the positive direct methods and/or positive sIgA antibody results from cervical or urethral specimens had an indication of current C. trachomatis infection.

Purpose: This study aimed to develop an improved method for the detection of bacterial SHV-type extended-spectrum β-lactamases (ESBLs). Materials and Methods: Our method was based on real-time polymerase chain reaction (PCR) in which the amplification of the product was monitored with a fluorescent probe. This method enabled the detection of bla _SHV genes with high degrees of sensitivity and specificity. Results: Based on ESBL phenotyping methods and bla gene DNA sequencing, we identified 240 bla genes from 662 Enterobacteriaceae isolated from clinical culture specimens. Of these 240 isolates, 26 had the bla _SHV-28 genotype and three had the bla _SHV-1 genotype. With our new real-time PCR assay, we detected 29 out of 29 bla _SHV genes in ESBL-producing isolates. Conclusion: This method represents a powerful tool for epidemiological studies of SHV ESBLs. Furthermore, it has potential for use in diagnostic microbiology.

Purpose: The purpose of the present study was to perform a molecular epidemiological survey by investigating the antibiotic resistance and the presence of known virulence factors in Enterococcus faecium isolates in Iran. The data collected from this study would allow us to control the spread and develop strategies for treatment of the enterococcal infections. Materials and Methods: In this study, 156 vancomycin-sensitive E. faecium (VSEF; 58) and vancomycin-resistant E. faecium (VREF; 98) samples were isolated from clinical specimen and sewage treatment plants (STPs). These isolates were screened for the presence of genes encoding for aggregation substance (asa1), cytolysin (cyl), enterococcal surface protein (esp), gelatinase (gelE) and hyaluronidase (hyl) by polymerase chain reaction (PCR). Results: Although significantly different, the results showed the presence of hyl and esp genes in both clinical (41 and 75%, respectively) and sewage (3.2 and 41%, respectively) isolates. Sensitivity of all isolates to seven antibiotics was examined. The results of the clinical isolates showed that the majority of esp positive isolates were also resistant to vancomycin, ciprofloxacin and erythromycin. Furthermore, cyl, gelE and asa1 were not found in either clinical or STP isolates. Finally, we determined the distinct types of isolates using Pulse Field Gel Electrophoresis (PFGE), which confirmed that most of the isolates were clonally unrelated. Conclusion: Our results demonstrated that higher number of the clinical E. faecium isolates carried virulence genes than the isolates from STP. Finally, the lack of the genes in clinical and STP isolates confirmed that these genes do not transfer horizontally.

Purpose: Surveillance of hospital-acquired infection (HAI), particularly device-associated infection (DAI), helps in determining the infection rates, risk factors, and in planning the preventive strategies to ensure a quality healthcare in any hospital. The present study was carried out to know the prevalence of DAI in a tertiary care teaching hospital of rural Gujarat. Materials and Methods: A prospective, site-specific surveillance of three common DAIs that is catheter-associated urinary tract infection (CA-UTI), IV-catheter-related bloodstream infection (IV-CRBSI), and ventilator-associated pneumonia (VAP) was carried out between July 2007 and April 2008, in different wards/ICUs. A surveillance plan, with guidelines and responsibilities of nurses, clinicians and microbiologist was prepared. Infection surveillance form for each patient suspected to have DAI was filled. The most representative clinical sample, depending on the type of suspected DAI, was collected using standard aseptic techniques and processed for aerobes and facultative anaerobes. All the isolates were identified and antimicrobial sensitivity testing performed as per CLSI guidelines. An accurate record of total device days for each of the indwelling devices under surveillance was also maintained. Data, collected in the prescribed formats, were analysed on monthly basis; and then, compiled at the end of the study. Descriptive analysis of the data was done and DAI rate was expressed as number of DAI per 1000 device days. Results: The overall infection rate for CA-UTI, IV-CRBSI, and VAP were found to be 0.6, 0.48, and 21.92 per 1000 device days, respectively. The organisms isolated were Staphylococcus aureus, CONS, Enterococci, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Escherichia coli. Conclusions: Duration of indwelling devices was found to be the major risk-factor for acquiring DAIs. Low DAI rate might have been due to use of antibiotics, often prophylactic. Active surveillance is quite a tedious and time-consuming process; however the outcome is useful in prevention and control of DAIs.

Purpose: Brain abscesses often present an aetiological dilemma. Microscopy is insensitive and culture techniques are time consuming. Hence, a new rapid technique in vitro Proton Magnetic Resonance Spectroscopy ( ¹ HMRS) was evaluated for its usefulness in the identification of aetiology of brain abscesses. Materials and Methods: A total of 39 pus specimens from brain abscesses were subjected to in vitro ¹ HMRS. These pus specimens were also processed by conventional culture methods. The spectral patterns generated by in vitro ¹ HMRS were further correlated with culture results. Results: Pus specimens which showed the presence of anaerobes on culture revealed the presence of multiplet at 0.9 ppm (100%), lactate-lipid at 1.3 ppm (100%), acetate at 1.92 ppm (100%) and succinate at 2.4 ppm (75%). Pus specimens that revealed the presence of facultative anaerobes on culture showed a pattern B, i.e., the presence of lactate-lipid at 1.3 ppm (100%), acetate at 1.92 ppm (88.88%) along with the multiplet at 0.9 ppm (100%). Pattern C was seen in aerobic infection which showed the presence of lactate-lipid at 1.3 ppm (100%) along with the multiplet at 0.9 ppm. Pus from two tuberculous abscesses showed the complete absence of multiplet at 0.9 ppm. Conclusions: We observed in this study that it was possible to differentiate bacterial and tuberculous brain abscesses using in vitro ¹ HMRS. Further, it was also possible to distinguish between aerobic and anaerobic brain abscesses on the basis of spectral patterns. In vitro ¹ HMRS of fungal and actinomycotic brain abscess are also presented for its unusual spectra.

Purpose: The purpose of the study was to determine the prevalence of Toxoplasma antibodies in selected immunocompetent and immunodeficient patients in and around Tirunelveli District of Tamil Nadu. Materials and Methods: This study was carried out from May 2006 to October 2007 in 175 immunodeficient and 175 immunocompetent patients. Serum samples were subjected into in-house IgG assay using indirect fluorescent antibody test (IFAT). Results: Out of 350 patients tested by IgG IFAT, 41 (11.71%) had antibodies for Toxoplasma gondii with a mean IFA titre of 43.42 ± 58.7 and the titre ranging from 1 : 16 to 1 : 256. Among the immunocompetent and immunodeficient groups, 19 patients (10.86%) and 22 patients (12.57%), respectively, had antibodies to T. gondii. Various risk factors associated within the study group was analysed and results were interpreted. Conclusions: The study has highlighted an overall seroprevalence of 11.71% with 12.57% in immunocompromised and 10.86% in immunocompetent patients respectively in a southern district, Tamil Nadu, which underlines the importance of screening of this parasite especially in the immunocompromised patients.

Characterisation of host immune response to hepatitis C virus (HCV) genotypes may have an important prognostic and therapeutic implication. Genotype-3 was more prevalent in the examined cohort and demonstrated a significantly higher response to combination therapy than genotype-1. Sustained virological response (SVR) was 94.74% in genotype-3 and 45.45% in genotype-1. The patients who achieved SVR reported higher levels of circulating T helper 1 cytokines in comparison to subjects with no SVR in both the studied groups. Besides providing local prevalence, our study might also assist in understanding the host immune mechanisms involved to achieve SVR during combination therapy in chronic HCV patients.

The aim of this study was to evaluate a nitrate reductase assay (NRA) for the direct detection of multidrug resistance (MDR) in Mycobacterium tuberculosis from 100 smear-positive sputum samples. The NRA results were compared with the reference proportion method for 100 sputum specimens for which comparable results were available. NRA results were obtained at day 7 for 61 specimens, results for 26 specimens were obtained at day 10, and the results for 13 specimens were obtained at day 14. Thus, 87% of NRA results were obtained in 10 days. NRA is a rapid, accurate, and cost-effective method for the detection of MDR in M. tuberculosis isolates as compared to the proportion method, which is time consuming. Therefore, NRA constitutes a useful tool for detection of tuberculosis drug resistance in low-resource countries with limited laboratory facilities due to its low-cost, ease of performance and lack of requirement of sophisticated equipment.

The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth) for the diagnosis of tuberculous meningitis (TBM) in cerebrospinal fluid (CSF). CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50) were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64%) out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI): 67-93%] and 36% specificity (95% CI: 57-98%) for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM). This coexistence may go undetected or potentially lead to erroneous reporting of results.

In this study, in vitro activity of tigecycline (TIG) and ertapenem (ERT) against clinical isolates of Brucella melitensis and the effect of different media on in vitro test results were investigated. The in vitro effects of TIG and ERT to 38 B. melitensis isolates were comparatively investigated in brucella agar and 5% sheep blood agar. MIC value of ERT was 0.032 μg/mL in 23 of 38 and 20 of 38 isolates on blood and brucella agar, respectively. Minimum inhibitory concentration values of TIG were substantially different ranging between 0.064-0.25 μg/mL on blood agar. However, MIC values of TIG were similar on brucella agar with 0.25 μg/mL in 15 of 38 isolates and 0.5 μg/mL in 10 of 38 isolates. In conclusion, although ERT and TIG were effective against B. melitensis isolates in vitro, further studies are needed in order to determine the use of these novel drugs in treatment of brucellosis.

The present study was carried out to determine the rates of high-level and low-level mupirocin resistance in Staphylococcus spp. (MuH and MuL) in southern India. A prospective study was carried out on Staphylococcus spp. isolated for a period of three months in the microbiology laboratory of an 800-bedded tertiary care hospital. One hundred sixty-seven non-duplicate Staphylococcus spp. isolated from different specimens were tested for mupirocin susceptibility using 5 and 200 μg discs and by agar dilution. Rates of MuH were found to be two percent in methicillin-resistant Staphylococcus aureus (MRSA) and 28% in methicillin-resistant coagulase-negative Staphylococcus spp. (MRCoNS). MuL strains may be still treated with mupirocin, while MuH strains require other treatment options for eradication, making prior screening and differentiation important.

Extended spectrum β-lactamases (ESBLs) have been observed in virtually all the species of family Enterobacteriaceae. Threat posed by antibiotic resistance because of ESBLs is more serious as a number of technical problems are associated with the detection of these enzymes. Although a number of detection methods have been designed for ESBLs, every method has its own benefits and shortcomings as well. In earlier days, isoelectric focusing (IEF) was used as the gold standard for ESBL detection. This study was undertaken to compare IEF with polymerase chain reaction, a method which has been extensively used for ESBL detection these days.

A total of 14 phenotypically similar clinical isolates of Klebsiella pneumoniae, resistant to multiple drugs including cefotaxime and ceftazidime, were isolated from blood of neonates admitted to neonatal intensive care unit (NICU) within a short span of 10 days. Alarmed at the possibility of occurrence of outbreak, a thorough investigation was done. Microbiological sampling of the NICU and labour room (LR) environment yielded 12 K. pneumoniae isolates. The presence of extended spectrum β-lactamase (ESBL) in the clinical and environmental strains was detected by double-disk synergy test (DDST), CLSI phenotypic confirmatory disk diffusion test (PCDDT) and E-test ESBL strips. Amp-C screen (disk) test was done to determine Amp-C β-lactamase production. 100% clinical strains, 57% NICU strains and 80% LR strains were ESBL positive. 57% clinical, 43% NICU and 20% LR strains were Amp-C screen positive. Polymerase chain reaction (PCR) of representative ESBL positive (10 clinical and 5 environmental) strains showed CTX gene and TEM and/or SHV gene in all. K. pneumoniae showing multiple mechanisms of drug resistance was responsible for the outbreak.

Perinatal listerial infection is the most common clinical syndrome caused by Listeria monocytogenes and includes abortion, still birth, neonatal sepsis, and meningitis. Reports of listeriosis from India are limited. Sub Himalayan . We report a case of neonatal listeriosis from Himachal Pradesh. A two-day-old full term male baby was referred from a peripheral hospital with fever listlessness, skin rash and non-acceptance of feed. Ceftriaxone was already started as an empirical therapy. Listeria monocytogenes was isolated from cerebrospinal fluid (CSF) and blood of the baby, and also from the genital tract of the mother. Unfortunately, the baby died before the preliminary report could be communicated.

This is an unusual report of isolation of Streptobacillus moniliformis from the blood of a male child with acute lymphoblastic leukaemia. No history of rat bite was there, but rats were present in the house. The possible source of infection may be food or water contaminated with rat excreta. Whether this bacteria can cause opportunistic infection in leukaemic patients, need to be evaluated further.

Empyema thoracis by Nocardia farcinica infection is uncommon. Here we report a rare and fatal infection in a 27-year-old HIV- seropositive male who presented with cough, expectoration, and breathlessness. Nocardia farcinica was isolated from sputum and pus from the pleural cavity. Confirmation of the isolate and minimum inhibitory concentrations (MIC) for various antibiotics was done at the Aerobic Actinomycetes Reference Laboratory, Centres for Disease Control and Prevention (CDC), Atlanta. Patient was treated with suitable antibiotics and antiretroviral drugs in spite of which he eventually succumbed to the disease.

Lymphatic filariasis is common in tropical countries and is endemic in India. Filariasis presenting with pleural effusion is an unusual presentation. Malignancy in association of filarial pleural effusion is extremely rare. We report a case of a 60-year-old female who presented with chest pain, loss of weight and breathlessness for a few months. Pleural fluid examination revealed malignant cells, along with microfilaria of Wuchereria bancrofti.

Human subcutaneous dirofilariasis is a rare infection caused by filarial worms of the genus Dirofilaria. The parasites are transmitted to man by mosquitoes and the infection is manifested as subcutaneous nodules. Excision of the lesion is both diagnostic and therapeutic. Hereby we report three cases of human subcutaneous dirofilariasis. The worms were sent to our department for identification over a period of four years (2006-2009). Of these three patients, two men and one woman were between 15 and 45 years of age. In two cases, the infection manifested as a nodule on face, in one case near lower eyelid and in the other on the cheek, while in the third case as an itchy nodule on the abdomen. It is emphasized that both clinicians and microbiologists should have an increased awareness of this entity and include dirofilariasis in the differential diagnosis of patients presenting with subcutaneous nodules.

A case of phaeohyphomycosis presenting as multiple subcutaneous abscesses in a young lady with deteriorating liver function was reported here. The lesion started as a solitary abscess in the neck, mimicking tuberculous cold abscess and rapidly involved the face, chest, arms, and legs within six months with ulceration and discharge of thick brownish foul smelling pus. Potassium hydroxide mount of pus from various sites revealed septate dematiaceous hyphae and pseudohyphae. Culture yielded pure growth of Exophiala spinifera. Tissue debridement was done along with initiation of antifungal therapy with ketoconazole. As liver function deteriorated, antifungal therapy was withdrawn after seven days. Patient expired three weeks after admission due to hepatic failure.

A 35-year-old male patient presented with complaints of redness, swelling around the eyelids, watering, and irritation in the right eye. At presentation his best-corrected visual acuity was 20/20 partial in the right eye. The tarsal conjunctiva of the upper eyelid showed injection with pseudomembrane. Underneath the pseudomembrane we noticed four motile larvae. The cornea showed an irregular cobweb-like mucous plaque adherent to the epithelium, with a clear stroma. The pseudomembrane was easily peeled-off under topical anaesthesia. The organisms were removed and identified as Oestrus ovis. Three days later the patient was comfortable and his visual acuity was 20/20 in the right eye.

A case of allergic bronchopulmonary aspergillosis (ABPA) is being described in a 52-year-old female patient who presented with a cavitary lesion on skiagram chest and simulating a lung abscess. Patient responded with the oral corticosteroid therapy with complete resolution of the initial radiographic abnormality. Despite various radiological presentations described in the literature, a lung abscess like presentation in ABPA is very rare and significant, because an early and correct diagnosis by the clinicians will help in early management of these cases to prevent the development of end-stage pulmonary fibrosis.

A patient with human immunodeficiency virus (HIV) infection presented with multiple cutaneous lesions on upper extremities, trunk, face and with ulcers involving oral mucosa. Histoplasma capsulatum was isolated in culture from scrapings from both cutaneous as well as oral mucosal lesions. The patient responded well initially to the treatment with Amphotericin B followed by itraconazole; however, lesions recurred after three months with the further deterioration of immune status of the patient indicated by decline in CD4 counts. The same treatment was restarted and the patient is still being followed-up.

A 58-year-old male diabetic who was operated for carcinoma larynx 4 years back was admitted with exertional dyspnoea and bilateral leg swelling for the past 2 years. Over the last 2 months, there was a progressive worsening of symptoms. Echocardiography done 2 years back showed pericardial effusion. Echo done during the current admission also showed pericardial effusion with preserved left ventricular function; cytological examination of the pericardial fluid showed larvae of Strongyloides stercoralis. He was treated with antinematodal drugs. A follow-up echo done at discharge showed no pericardial effusion and the patient was completely asymptomatic. To our knowledge, this is the first reported case of Strongyloides pericardial effusion in a diabetic patient.

We report a case of thyroid abscess caused by Scedosporium apiospermum in a patient with cirrhosis of liver and autoimmune haemolytic anaemia. To date, there are no reports of isolation of this fungus from thyroid abscess.