Clonal clusters and virulence factors of methicillin-resistant Staphylococcus Aureus: Evidence for community-acquired methicillin-resistant Staphylococcus Aureus infiltration into hospital settings in Chennai, South India

Background and Objective: Staphylococcus aureus is one of the major pathogens of nosocomial infections as wells as community-acquired (CA) infections worldwide. So far, large-scale comprehensive molecular and epidemiological characterisation of S. aureus from very diverse settings has not been carried out in India. The objective of this study is to evaluate the molecular, epidemiological and virulence characteristics of S. aureus in both community and hospital settings in Chennai, southern India. Methods: S. aureus isolates were obtained from four different groups (a) healthy individuals from closed community settings, (b) inpatients from hospitals, (c) outpatients from hospitals, representing isolates of hospital–community interface and (d) HIV-infected patients to define isolates associated with the immunocompromised. Antibiotic susceptibility testing, multiplex polymerase chain reactions for detection of virulence and resistance determinants, molecular typing including Staphylococcal cassette chromosome mec (SCCmec) and agr typing, were carried out. Sequencing-based typing was done using spa and multilocus sequence typing (MLST) methods. Clonal complexes (CC) of hospital and CA methicillin-resistant S. aureus (MRSA) were identified and compared for virulence and resistance. Results and Conclusion: A total of 769 isolates of S. aureus isolates were studied. The prevalence of MRSA was found to be 7.17%, 81.67%, 58.33% and 22.85% for groups a, b, c and d, respectively. Of the four SCCmec types (I, III, IV and V) detected, SCCmec V was found to be predominant. Panton-Valentine leucocidin toxin genes were detected among MRSA isolates harbouring SCCmec IV and V. A total of 78 spa types were detected, t657 being the most prevalent. 13 MLST types belonging to 9 CC were detected. CC1 (ST-772, ST-1) and CC8 (ST238, ST368 and ST1208) were found to be predominant among MRSA. CA-MRSA isolates with SCCmec IV and V were isolated from all study groups including hospitalised patients and were found to be similar by molecular tools. This shows that CA MRSA has probably infiltrated into the hospital settings.

Paediatric leptospirosis: A population based case–control study from Chennai, India.

The surveillance in Chennai identified 134 children and 443 adults clinically suspected for leptospirosis. Of these, 35 (26.1%) children and 118 (26.6%) adults had laboratory confirmed diagnosis for leptospirosis. The paediatric leptospirosis exhibited a higher frequency of classic features of Weil’s disease. The prevalent serovar encountered was Icterohaemorrhagiae with no difference in the pattern of infecting serovars between the two groups. Further, confirmation of diagnosis was achieved by polymerase chain reaction (PCR) with a positivity of 28.4% (specificity 96%). Univariate analysis showed significant association of paediatric leptospirosis with rat infestation (odds ratio 87.4). Thus, PCR facilitates early diagnosis of febrile illness among paediatric cases.

HIV reverse transcriptase gene mutations in anti‑retroviral treatment naïve rural people living with HIV/AIDS.

This study is designed to find out the mutational variations of reverse transcriptase (RT) gene of HIV, after the traditional drug usage among anti‑retroviral therapy naïve rural people living with HIV/AIDS. HIV Reactive patients, who were exposed for indigenous medicines such as Siddha, Ayurveda etc., for a minimum period of 6 months were taken for this study. Among 40 patients, two samples (5.55%) demonstrated high‑level mutational resistance variations for nucleoside RT inhibitor (NRTI) and non‑NRTI. The predominant polymorphisms detected were K122E (91.7%), V60I (91.7%), V35T (89%), Q207E (89%), D177E (89%), T200A (86.1%), S48T (83.33%), K173A (80.6%).

Complete sequencing of an IncX3 plasmid carrying blaNDM-5 allele reveals an early stage in the dissemination of the blaNDM gene.

Purpose: The aim of the present study was to perform molecular characterisation of the blaNDM plasmids and to understand the mechanism of its spread among pathogenic bacteria. Materials and Methods: Seventy-six non-repetitive carbapenem-resistant isolates which were collected during Nov 2011 to April 2013 from four hospitals in Chennai were analyzed for the presence of the blaNDM gene by PCR. Further, the genetic context of the blaNDM gene was analyzed by PCR specifi c to ISAba125 and bleMBL gene. One of the blaNDM plasmid was completely sequenced in the Illumina HiSeq platform. Results: Twenty-three isolates consisting of 8 Escherichia coli, 8 Klebsiella pneumoniae, 3 Klebsiella oxytoca, 3 Acinetobacter baumanii and 1 Pseudomonas aeruginosa were found to carry the blaNDM gene. In 18 isolates the blaNDM gene was associated with a bleMBL gene and the ISAba125 element. The complete sequencing of pNDM-MGR194 revealed an IncX3 replication type plasmid, with a length of 46,253 bp, an average GC content of 47% and 59 putative ORFs. The iteron region contained the blaNDM5 gene and the bleMBL, trpF and dsbC genes downstream and an IS5 inserted within the ISAba125 element upstream. Conclusion: This is the fi rst report where the blaNDM gene insertion in a plasmid is not accompanied by other resistance gene determinants. These observations suggest that the IncX3 plasmid pNDM-MGR194 is an early stage in the dissemination of the blaNDM.

Healthcare Professionals’ Perspective of Support from Public Health Department: A Study in the Primary Health Centers of Tamil Nadu.

Background: Perceived organizational support (POS) measures the employee perception of support from the organization and helps to understand the employer-employee relationship better. The aim of the study is to understand the healthcare professionals’ perception of support from public health department and to examine thehealthcare professionals’ difference in perception of support across various demographics variables. Methods: A survey is conducted among healthcare professionals including medical offi cers and staff nurses working in primary health centres (PHC) in the state of Tamilnadu. In the study, public health department refers to the Directorate of Public Health and Preventive Medicine. Mann Whitney U test and Kruskal Wallis H test are used to assess the difference in POS across demographic variables. Results: The results of the study indicated that the healthcare professionals perceive a moderate levelof support from public health department. The healthcare professionals’ POS is found to differ across their age, role and total work experience but not across gender and work experience in the current PHC. Further, the POS of healthcare professionals is found to differ with respect to PHC location and does not differ with respect to PHC type. Conclusion: The results emphasize the need to enhance POS of healthcare professionals. The health policy makers and mangers have to consider the difference in POS of healthcare professionals and make amendments in the human resource policies related to selection and training.

Impact of organic rich diet on gut enzymes, microbes and biomass of earthworm, Eudrilus eugienea.

Vermitechnology provides scope and opportunities in the field of Biotechnology. The sudden decline in earthworm biomass may mainly be due to the over use of chemicals. Steps had been taken to enhance the production of biomass of earthworms by providing organic wastes rich in major organic constituents such as cereals, pulses and skin of chick. Earthworms (Eudrilus eugienea) were fed with organic constituents individually, and in combination of organic rich diet. The biomass of the earthworm was steadily increasing in the individual treatment was found when fed with organic rich diet at 10, 20 and 30 days respectively 09.987, 13.569 and 18.212. The bacterial counts in the gut of earthworms were 543 x 105 CFU ml-1. The bacteria identified were Bacillus spp., Lactobacillus spp and Flavobacterium spp. Enzymes screened in the gut were amylase, endoglucanase, cellulase, sucrase and protease. From the present investigation, it was found that the organic rich diet is the ideal medium in which the biomass of earthworms are high, their enzymatic activity was also high with variety of microbes which will enhance the efficiency of the soil.

Trichosporon inkin, an unusual agent of fungal sinusitis: A report from south India.

The aetiology of fungal sinusitis is diverse and changing. Aspergillus species has been the most common cause for fungal sinusitis, especially in dry and hot regions like India. Trichosporon species as a cause for fungal sinusitis has been very rarely reported the world over. Here, we report a rare case of allergic fungal sinusitis caused by Trichosporon inkin in a 28-year-old immunocompetent woman. Bilateral nasal obstruction, nasal discharge and loss of smell were her presenting complaints. Diagnostic nasal endoscopy showed bilateral multiple polyps. Functional endoscopic sinus surgery was performed and many polyps were removed. Based on mycological and histopathological studies, the pathogen was identified as T. inkin.

Identification of plasmid-mediated quinolone resistance genes qnrA1, qnrB1 and aac(6')-1b-cr in a multiple drug-resistant isolate of Klebsiella pneumoniae from Chennai. Indian Journal of Medical Microbiology. 2011 Jul-Sept; 29(3): 262-268.

Purpose: Resistance to fluoroquinolones, a commonly prescribed antimicrobial for Gram-negative and Gram-positive microorganisms, is of importance in therapy. The purpose of this study was to screen for the presence of Plasmid-Mediated Quinolone Resistance (PMQR) determinants in clinical isolates of Klebsiella pneumoniae. Materials and Methods: Extended-Spectrum Beta-Lactamase (ESBL) isolates of K. pneumoniae collected during October 2009 were screened by the antimicrobial susceptibility test. The plasmids from these isolates were analysed by specific Polymerase chain Reaction (PCR) for qnrA, qnrB and aac(6')-1b. The amplified products were sequenced to confirm the allele. Results: Our analysis showed that 61% out of the 23 ESBL K. pneumoniae isolates were resistant to ciprofloxacin and 56% to levofloxacin. The PMQR was demonstrated by transforming the plasmids from two isolates P12 and P13 into E. coli JM109. The PMQR gene qnrA was found in 16 isolates and qnrB in 11 isolates. The plasmid pKNMGR13 which conferred an minimum inhibitory concentration (MIC) of more than 240 ΅g/ml in sensitive E. coli was found to harbour the qnrA1 and qnrB1 allele. Furthermore, the gene aac(6')-1b-cr encoding a variant aminoglycoside 6'-N Acetyl transferase which confers resistance to fluoroquinolones was found in the same plasmid. Conclusions: Our report shows the prevalence of PMQR mediated by qnrA and qnrB in multidrug-resistant K. pneumoniae isolates from Chennai. A multidrug-resistant plasmid conferring high resistance to ciprofloxacin was found to harbour another PMQR gene, aac(6')-1b-cr mutant gene. This is the first report screening for PMQR in K. pneumoniae isolates from India.

Identification of plasmid-mediated quinolone resistance genes qnrA1, qnrB1 and aac(6')-1b-cr in a multiple drug-resistant isolate of Klebsiella pneumoniae from Chennai.

Purpose: Resistance to fluoroquinolones, a commonly prescribed antimicrobial for Gram-negative and Gram-positive microorganisms, is of importance in therapy. The purpose of this study was to screen for the presence of Plasmid-Mediated Quinolone Resistance (PMQR) determinants in clinical isolates of Klebsiella pneumoniae. Materials and Methods: Extended-Spectrum Beta-Lactamase (ESBL) isolates of K. pneumoniae collected during October 2009 were screened by the antimicrobial susceptibility test. The plasmids from these isolates were analysed by specific Polymerase chain Reaction (PCR) for qnrA, qnrB and aac(6')-1b. The amplified products were sequenced to confirm the allele. Results: Our analysis showed that 61% out of the 23 ESBL K. pneumoniae isolates were resistant to ciprofloxacin and 56% to levofloxacin. The PMQR was demonstrated by transforming the plasmids from two isolates P12 and P13 into E. coli JM109. The PMQR gene qnrA was found in 16 isolates and qnrB in 11 isolates. The plasmid pKNMGR13 which conferred an minimum inhibitory concentration (MIC) of more than 240 ΅g/ml in sensitive E. coli was found to harbour the qnrA1 and qnrB1 allele. Furthermore, the gene aac(6')-1b-cr encoding a variant aminoglycoside 6'-N Acetyl transferase which confers resistance to fluoroquinolones was found in the same plasmid. Conclusions: Our report shows the prevalence of PMQR mediated by qnrA and qnrB in multidrug-resistant K. pneumoniae isolates from Chennai. A multidrug-resistant plasmid conferring high resistance to ciprofloxacin was found to harbour another PMQR gene, aac(6')-1b-cr mutant gene. This is the first report screening for PMQR in K. pneumoniae isolates from India.

Subcutaneous phaeohyphomycosis due to Exophiala spinifera in an immunocompromised host.

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.