Gulnaz Bashir

Bipolaris hawaiiensis Keratomycosis and Endophthalmitis

Following trauma with rice stalk to the left eye, corneal ulcer with abscess and hypopyon developed in an immunocompetent male. Direct examination of the corneal scrapings revealed septate, branched fungal hyphae. Bipolaris hawaiiensis was isolated after culture on blood agar and Sabouraud dextrose agar. Because of delay in diagnosis and appropriate treatment, the patient developed endophthalmitis needing evisceration.

Clinical profile and surgical outcome for pulmonary aspergilloma: nine year retrospective observational study in a tertiary care hospital.

BACKGROUND The indications and the outcome of surgery for pulmonary aspergilloma remain highly controversial. This retrospective observational study was conducted to study the clinical profile, indications, post-operative complications and long term outcome of patients having pulmonary aspergilloma. METHODS From January 2000 to October 2008, 52 patients underwent surgery for pulmonary aspergilloma at our tertiary care institute. RESULTS The group consisted of 32 males and 20 females with a mean age of 39.3 ± 11.2 years. The most common indication for surgery was hemoptysis (96.15%). The underlying lung diseases were tuberculosis (75%), bronchiectasis (5.76%), and lung abscess (5.76%). In one patient (2%), concomitant ruptured lung hydatid cyst and an aspergilloma was present. The procedures performed were lobectomy (n = 43), bilobectomy (n = 3). pneumonectomy (n = 3), segmental resection (n = 3). The post-operative mortality was 1.92% (one patient). Overall complications occurred in 12 (23.07%) patients. The complications included prolonged air leak (n = 6), bleeding (n = 3), empyema (n = 1), repeated pneumothorax (n = 1), and wound dehiscence (n = 1). The mean follow-up period was 38 ± 18.6 months. There was no recurrence of disease or hemoptysis. CONCLUSION Pulmonary aspergilloma is common in developing countries like India in which there is high prevalence of pulmonary tuberculosis. Surgical resection of pulmonary aspergilloma is effective in preventing recurrence of symptoms including hemoptysis. We recommend early surgical resection of symptomatic aspergilloma with reasonable complications. Pre-operative preparation of the patients, meticulous surgical technique and post-operative chest physiotherapy reduces the rate of complications. Complications may still occur and are largely related to the underlying lung pathology; however, the long term outcome is good.

Sepsis due to linezolid resistant Staphylococcus cohnii and Staphylococcus kloosii: First reports of linezolid resistance in coagulase negative staphylococci from India

Linezolid, a viable alternative to vancomycin against methicillin resistant staphylococcal isolates, has been in use for a decade around the globe. However, resistance against staphylococci remains extremely rare and unreported from most of the Asian countries. Herein, we report two cases of linezolid resistant, coagulase negative staphylococcal sepsis for the first time from India. The first case was an 18-year-old burn patient, who, after a major graft surgery, landed in sepsis, and linezolid resistant Staphylococcus cohnii with an minimum inhibitory concentration (MIC) of >256 μg/ml by both broth microdilution and Etest, was isolated from multiple blood cultures. The second patient was a 60-year-old male with an intracranial bleed and sepsis, from whose blood cultures, linezolid resistant Staphylococcus kloosii was repeatedly isolated. Linezolid MIC was >32 μg/ml by broth microdilution and >16 μg/ml by Etest.

Pulmonary pseudallescheriasis in a patient with healed tuberculosis

Pulmonary pseudallescheriasis in an immunocompetent patient without a pre-existing cavity or cyst is a rare phenomenon. We report a case of invasive pulmonary pseudallescheriasis in a lobectomised patient treated for tuberculosis. Filamentous fungi with pyriform conidia were seen in the bronchoalveolar lavage fluid .The fungus was identified as Pseudallescheria boydii on culture.

Nasal carriage of Methicillin-resistant Staphylococcus aureus among healthy population of Kashmir, India.

BACKGROUND Nasal colonisation with community acquired methicillin resistant Staphylococcus aureus (CA-MRSA) is being increasingly reported, especially in places where people are in close contact and where hygiene is compromised. The aim of this study was to find out prevalence of methicillin resistant S.aureus (MRSA) colonising anterior nares of healthy subjects. MATERIALS AND METHODS Nasal swabs of healthy subjects were collected aseptically and cultured using standard microbiological protocols. Antibiotic susceptibility was done by Kirby-Bauer disc diffusion method according to CLSI guidelines. Methicillin resistance was detected by cefoxitin disc diffusion method and confirmed by minimum inhibitory concentration (MIC) and amplification of mecA gene by PCR. Strain typing of MRSA strains was done by PFGE. RESULTS Out of 820 samples, S.aureus was isolated from 229 (27.92%) subjects. Of the 229 isolates, 15 were methicillin resistant. All S. aureus isolates were susceptible to vancomycin. Nasal carriage of MRSA was found to be 1.83% among healthy population. The isolates were found to be polyclonal by PFGE analysis. CONCLUSION High prevalence of MRSA is a cause of concern and strategies to interrupt transmission should be implemented.

Fungemia caused by Verticillium species in an immunocompromised child

The incidence of fungal infections is increasing due to immunocompromised states. We report a case of fungaemia due to a rare fungus - Verticillium, in a 6 year old child diagnosed as a case of acute lymphoblastic leukaemia- L1 with high grade fever. The patient was treated with amphotericin B with a good clinical response.

Rapid drug-susceptibility testing of Mycobacterium tuberculosis clinical isolates to first-line antitubercular drugs by nitrate reductase assay: A comparison with proportion method

Objective/background: Early initiation of therapy in patients with tuberculosis is imperative for its control. Conventional methods of susceptibility testing such as the proportion method (PM) require visual detection and counting of colonies that takes up to 6 weeks. Rapid and simple phenotypic methods that have been endorsed by the World Health Organization can serve as alternatives. Methods: In this study, we evaluated the colorimetric nitrate reductase assay, which utilizes the detection of nitrate reduction as an indicator of growth much earlier compared with PM (within 7–14 days). The susceptibility of 75 clinical isolates of Mycobacterium tuberculosis to four first-line antitubercular drugs was tested by nitrate reductase assay and compared with the standard PM. In this assay, inoculation was done on both drug-free and drug-containing Löwenstein–Jensen medium containing sodium nitrate. After incubation for 7–14 days, reduction to nitrite was taken as an indicator of growth, which was detected by color change on addition of Griess reagent. Results: Agreement between nitrate reductase assay and PM was 100% for rifampicin, 97.30% for isoniazid, 93.30% for streptomycin, and 98.60% for ethambutol. Cost/isolate with this assay was found to be approximately two times lesser than that of PM. All results were obtained in 7–14 days by nitrate reductase assay, which was significantly rapid compared with 42 days taken for obtaining results by PM. Conclusion: Nitrate reductase assay can be used as a rapid and inexpensive method for drug-susceptibility testing of M. Tuberculosis for first-line antitubercular drugs without compromising accuracy of standard methods.

Molecular identification of Candida species isolated from cases of neonatal candidemia using polymerase chain reaction-restriction fragment length polymorphism in a tertiary care hospital

Context: Candida spp. is an emerging cause of bloodstream infections worldwide. Delay in speciation of Candida isolates by conventional methods and resistance to antifungal drugs in various Candida species are responsible for the increase in morbidity and mortality due to candidemia. Hence, the rapid identification of Candida isolates is very important for the proper management of patients with candidemia. Aims: The aim was to re-evaluate the identification of various Candida spp. by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) and to evaluate the accuracy, speed, and cost of phenotypic methodology versus PCR-RFLP. Settings and Design: Hospital-based cross-sectional study. Materials and Methods: Ninety consecutive clinical isolates of seven Candida species, isolated from blood of neonates and identified by routine phenotypic methods, were re-evaluated using universal primers internal transcribed spacer 1 (ITS1) and ITS4 for PCR amplification and Msp I restriction enzyme for RFLP. Statistical Analysis Used: Kappa test for agreement. Results: The results of PCR-RFLP were 100% in agreement with those obtained using conventional phenotypic methods. Identification could be achieved within 3 work days by both the methods. Our routine methods proved to be cost effective than PCR-RFLP. Conclusions: We can continue with our routine phenotypic methods and PCR-RFLP can be used for periodic quality control or when conventional methods fail to identify a species.

Predominance of Central Asian and European families among Mycobacterium tuberculosis isolates in Kashmir Valley, India

BACKGROUND As there are no data available regarding the strains of Mycobacterium tuberculosis circulating in Kashmir Valley, India, the current study aimed at describing the genetic diversity of M. tuberculosis strains in this region, by spoligotyping and 12-locus-based MIRU-VNTR typing (Mycobacterial Interspersed Repetitive Unit-Variable Number Tandem Repeat). METHODS Sputa from 207 smear positive cases with newly diagnosed pulmonary tuberculosis were subjected to culture for M. tuberculosis. Eighty-five isolates confirmed as M. tuberculosis were subjected to drug susceptibility testing and molecular typing by spoligotyping and MIRU-VNTRs. RESULTS Drug susceptibility results of 72 isolates revealed 76.3% as fully sensitive while 5.5% as multidrug resistant (MDR). Spoligotyping of 85 isolates detected 42 spoligotypes with 50 isolates (58.8%) clustered into seven spoligotypes. SIT26/CAS1_Del was the major spoligotype (23, 27%) followed by SIT127/H4 (12, 14.1%); CAS lineage (37.6%) was predominant, followed by Haarlem (25.8%) and ill-defined T clade (23.5%). MIRU-VNTR analysis displayed 82 MIRU patterns from 85 strains, including 3 small clusters and 79 unique. MIRU 26 was found to be the most discriminatory locus. CONCLUSIONS Kashmir Valley has CAS as the predominant lineage of M. tuberculosis similar to the rest of the Indian sub-continent, while it is peculiar in having Euro American lineages such as Haarlem and ill-defined T clade.

Molecular Identification of Candida dubliniensis among Candida albicans Isolated from Oral Cavity of Cancer Patients using PCR-RFLP, in a Tertiary Care Hospital in Kashmir, India

Aims: To retrospectively evaluate 186 stock strains of C. albicans strains isolated from oral cavity of HIV negative patients with various malignancies for the presence of C. dubliniensis isolates among them by PCR-RFLP. Place and Duration of Study: Department of Microbiology, Sher-i-Kashmir Institute of Medical Sciences, Srinagar, between October 2013 and October 2014. Methodology: This study included 186 stock strains of C. albicans tentatively identified by phenotypic methods like germ tube formation in human serum, colony color on chromogenic Original Research Article Jan et al.; BMRJ, 14(2): 1-7, 2016; Article no.BMRJ.25465 2 candida differential agar, characteristic morphology on corn meal agar and assimilation of sugars isolated from HIV negative patients with various malignancies. DNA extraction was performed by chemical method. PCR amplification of ITS1-5.8S-ITS2 rDNA region was achieved using the ITS1 and ITS4 primer pairs which amplify the ITS region of both species, providing a single PCR product of expected size (540 bp). There is no visible difference between these two species with regard to their ITS PCR products. Digestion of amplified products was performed by using restriction enzyme BlnI (AvrII) which cleaves DNA where there is a CCTAGG sequence. The products of digestion generate one band of 540 bp for C. albicans, and two bands of 200 bp and 340 bp for C. dubliniensis because BlnI has one recognition site within the ITS region of C. dubliniensis, whereas none within that of C. albicans. Results: Of the 186 isolates tested, no C. dubliniensis was found by PCR-RFLP. Conclusion: Our results of not finding C. dubliniensis in this subset of patients support the need for further investigations into the prevalence of this species among other clinical samples and other susceptible patient populations.