Pathology, microbiology, blood bank and other diagnostic laboratories generate sizable amount of biomedical waste (BMW). The audit of the BMW is required for planning proper strategies. The audit in our laboratory revealed 8 kgs anatomical waste, 600 kgs microbiology waste, 220 kgs waste sharps, 15 kgs soiled waste, 111 kgs solid waste, 480 litres liquid waste along with 33000 litres per month liquid waste generated from labware washing and laboratory cleaning and 162 litres of chemical waste per month. Section wise details are described in the text. Needle sharps are collected in puncture proof containers and the needles autoclaved before sending to needle pit. The glass forms the major sharp category and is disinfected with hypochlorite before washing/recycling. All microbiology waste along with containers/plates/tubes are autoclaved before recycling/disposal. The problem of formalin fixed anatomical waste as histology specimens is pointed out. The formalin containing tissues cannot be sent for incineration for the fear of toxic gas release and the guidelines by the Biomedical waste rule makers need to be amended for the issue. The discarded/infected blood units in blood bank need to be autoclaved before disposal since chemical treatments are difficult or inefficient. The liquid waste management needs more attention and effluent treatment facility needs to be viewed seriously for hospital in general. The segregation of waste at source is the key step and reduction, reuse and recycling should be considered in proper perspectives.

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PURPOSE: To evaluate the performance of 65 kDa antigen based PCR assay in clinical samples obtained from pulmonary and extrapulmonary cases of tuberculosis. METHODS: One hundred and fifty six samples were processed for detection of Mycobacterium tuberculosis by ZN smear examination, LJ medium culture, BACTEC radiometric culture and PCR tests. RESULTS: A significant difference was seen in the sensitivities of different tests, the figures being 74.4% for PCR test, 33.79% for ZN smear examination, 48.9% for LJ culture and 55.8% for BACTEC culture (P<0.05). However, there was no significant difference (P>0.05) as far as specificity of different tests was concerned. PCR test sensitivity in pulmonary and extrapulmonary clinical samples were 72.7% and 75.9% respectively and found to be significantly higher (P<0.05) when compared with those of other tests. The mean detection time for M.tuberculosis was 24.03 days by LJ medium culture, 12.89 days by BACTEC culture and less than one day by PCR test. CONCLUSIONS: PCR is a rapid and sensitive method for the early diagnosis of pulmonary and extrapulmonary tuberculosis.

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In the present report, we have analysed C.trachomatis infection and HIV positivity among patients (n-143) who attended the STD clinic at the Institute of STDs, Government General Hospital, Chennai. HIV positivity rate was significantly high among those with chlamydial infection than in those without chlamydial infection (29.5% (13/44) vs. 11.1% (11/99); p<0.05). The results of the present study suggest the association between C.trachomatis and HIV infections and reinforce the need for routine screening for C.trachomatis as a necessary intervention to reduce the burden of chlamydial diseases and to reduce the risk of HIV and its spread in India.

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The objective of this study was to investigate screening methodologies, to detect Staphylococcus aureus strains with decreased susceptibility to vancomycin. Three methods were used to screen 160 Staphylococcus aureus clinical isolates along with ATCC quality control strains. Subsequently, MIC of all these 160 strains were determined by NCCLS methodology. The MIC of all the 160 clinical isolates was < 4µg/mL and were classified as vancomycin susceptible by NCCLS criteria but 23 strains were positive by Hiramatshu method, two grew on MHA (5µg/mL vancomycin) while CDC method correctly identified no vancomycin intermediate S.aureus (VISA) or vancomycin resistant S.aureus (VRSA) strains with reference to there MIC. CDC method was found to be the most appropriate screening methodology for detection of VISA or VRSA for diagnostic laboratories.

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Environmental samples were subjected to determine frequency of occurrence of pathogenic campylobacters in the environment. The antimicrobial susceptibility of the isolates was tested to evaluate the level of antibiotic sensitive campylobacters in the environment of investigation. In all, 70 Campylobacter isolates were obtained from water and domestic animal faeces samples using Kapadnis-Baseri device and antimicrobial susceptibility of them was determined by disc diffusion test and E- test. The results indicated that all the isolates of Campylobacter were sensitive to ciprofloxacin and resistant to cefotaxime, cephalexin and ampicillin. Lowest MIC values were observed for ciprofloxacin and gentamicin (2µg/mL) and highest MIC values for ampicillin and chloramphinicol (256µg/mL). In general, pathogenic Campylobacter spp. were prevalent in large numbers in the environment, however, they were sensitive to ciprofloxacin.

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Diphtheroids or "coryneform" bacilli are usually considered to be nonpathogenic "normal flora" of human skin and mucous membranes. Because bacterial cultures are frequently contaminated with these organisms the correct diagnosis and treatment may be delayed by the failure to recognize serious infections caused by them. Few confirmed cases of orthopaedic infections due to Arcanobacterium haemolyticum infection have been reported, partly because of inadequate identification of this bacterium. We report a case of septic arthritis due to A. haemolyticum.

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This report describes the use of the MTT-reduction and Evan's blue-staining tests for the assessment of the viability and morphological integrity, respectively, of rhinosporidial endospores after exposure to sera from rhinosporidial patients with high titres of anti-rhinosporidial antibody. Sera from three patients, with nasal, ocular and disseminated rhinosporidiosis respectively were used, with human serum without anti-rhinosporidial antibody for comparison, with or without added fresh guinea pig serum as a source of complement. All four sera tested, with or without guinea-pig serum, had no effect on the morphological integrity or the viability of the endospores and it is suggested that anti-rhinosporidial antibody has no direct protective role against the endospores, the infective stage, in rhinosporidiosis. This finding is compatible with the occurrence of chronicity, recurrence and dissemination that are characteristic of rhinosporidiosis despite the presence of high titres of anti-rhinosporidial antibody in patients with these clinical characteristics. The possible occurrence of humoral mechanisms of immunity that involve anti-rhinosporidial antibody with cells such as leucocytes and NK cells, in vivo, cannot yet be discounted, although the presence of high titres of anti-rhinosporidial antibody in patients with chronic, recurrent and disseminated lesions might indicate that such antibody is non-protective in vivo.

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PURPOSE: Porins are outer membrane protein (OMP) that form water filled channels that permit the diffusion of small hydrophilic solutes like -lactam antibiotics across the outer membrane. Two major porins that facilitate diffusion of antimicrobials have been described in Klebsiella spp. and Escherichia coli. The present study was carried out to examine the role of porins among Extended Spectrum -Lactamase (ESBL) and AmpC -Lactamase positive strains of Klebsiella spp. and E.coli. METHODS: Preparation of OMP from phenotypically characterized clinical isolates K.pneumoniae and E.coli and the separation of the proteins by sodium dodecyl sulfate - polyacrylamide gel electrophoresis were performed as per a previously described procedure. RESULTS: OMP analysis revealed that cefoxitin and ceftazidime resistance was mediated by loss of a porin Omp K35 in the isolates of K.pneumoniae and E.coli. CONCLUSIONS: Loss of porin mediated resistance mechanism against cefoxitin was observed among the multidrug resistant K.pneumoniae and E.coli.

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An unusual case of loiasis from Assam is reported here. Loa loa is a subcutaneous filarial parasite of man and is transmitted to humans by chrysops flies. The patient presented with foreign body sensation and visual disturbances of the right eye. Examination revealed a white coiled structure in the cornea.. Routine blood and other investigations were within normal limits. A live adult worm was extracted and identity was confirmed by microscopy to be Loa loa. Patient was treated with diethylcarbamazine and steroid. We found this case interesting as the worm was present in the anterior chamber - an unusual site and there were no other positive findings besides the lone worm.

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PURPOSE: To test the immunogenicity of the WHO recommended "2-2-2-0-1-1" post-exposure rabies vaccination regimen in Indian subjects to determine the feasibility of replacing crude sheep brain nerve tissue rabies vaccine with modern tissue culture rabies vaccine at major anti-rabies treatment centers throughout India. METHODS: Purified chick embryo cell vaccine (PCECV) was administered in the dosage of 0.1mL per site to 53 Indian subjects. RESULTS: All subjects produced rabies antibodies above 0.5IU/mL by day 14 post-vaccination. Only minor adverse reactions including swelling (6.6%), erythema (5.4%) and pain (1.4%) were observed for which no treatment was required. CONCLUSIONS: This study demonstrated that PCECV is safe and highly immunogenic in Indian subjects when administered intradermally as 0.1mL/site using the "2-2-2-0-1-1" post-exposure regimen.

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Colletotrichum dematium has been rarely reproted from India before. The present case, a farmer, developed peripheral corneal ulcer five days following trauma with plant. At presentation his visual acuity was 6/60 (unaided) and 6/24P with pinhole. Slit lamp and fluorescent stain examination revealed paracentral corneal ulcer with irregular margins, stromal infiltration and multiple epithelial defects. Microbiological examination of corneal samples confirmed the initial diagnosis of fungal corneal ulcer and the fungus was identified as C.dematium. Patient was treated with topical natamycin and ciprofloxacin. Patient left against medical advice and was lost to follow up. This report emphasizes that Colletotrichum keratitis may not be rare. Early diagnosis may help in institution of specific therapy early in the disease.

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Hepatitis B virus (HBV) is the most important causative agent of blood borne hepatitis in humans. Hepatitis D Virus (HDV) infection occurs either as a coinfection or superinfection in HBV carriers. Hepatitis C virus (HCV) is the major cause of transfusion non-A, non-B hepatitis and continues to be a major cause of human liver disease throughout the world. The present study was conducted on 70 clinically diagnosed cases of viral hepatitis to study the prevalence of parenterally transmitted viral hepatitis. The serum samples were tested for HBsAg, HBeAg, IgM anti-HBc, anti-HBe, anti-HCV and anti-HDV using separate ELISA kits. Of the 70 serum samples tested, 28(40%) were positive for HBsAg out of which 3 (4.28%) were positive for HBeAg also. Five (7.1%) of the HBsAg positive cases tested positive for IgM anti-HBc also. HBsAg alone was found in 17(24.28%) cases. The prevalence of anti-HCV was 3 (4.28%) in 70 cases. Thus early screening of clinically diagnosed cases of viral hepatitis is essential for establishing diagnosis and treatment to prevent long term sequelae.

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Hepatitis viral infections are important causes of morbidity and mortality in haemodialysis patients. One hundred and thirty four patients attending haemodialysis unit were screened for the presence of HBV and HCV infections. Eight (5.9%) patients were HCV positive while two (1.4%) patients had HBV infection. A dual infection with both the viruses was observed in five patients (3.7%).

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Three hundred and forty serum samples collected from women of child bearing age, without any clinical evidence of cytomegalovirus (CMV) infection, were screened for the presence of IgG antibodies against CMV by ELISA test. The IgG antibodies were detected in 297 which gave prevalence rate of 87.4%. Significantly higher prevalence rates (p < 0.001) were observed with increasing age and with increase in parity. There was significant difference in the antibody prevalence in different socioeconomic groups. Seroprevalence rate was also found to be more in women from rural area than those from urban area, although the difference was statistically not significant (p > 0.05). Marital status showed no impact upon the seroprevalence of IgG antibodies in women.

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Intestinal Heterophyid infections are almost unknown in India with only one documented case report. We report a case of six year old child with diarrhoea without any other associated gastrointestinal symptoms. Examination of the faeces revealed eggs of Metagonimus yokogawai. However, the patient left against medical advice and was lost to follow up. The case is being reported because of its rarity.

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