Characterization of leptospires using V3 region of 16S rDNA by denaturing gradient gel electrophoresis : A case study.

Serological and molecular characterization of Leptospiral isolates helps us to identify serovar, which is useful, for epidemiological study. Serological characterization is tedious and requires a panel of monoclonal antibodies and expertise to read the results. This study is a preliminary work to evaluate the usefulness of Denaturing Gradient Gel Electrophoresis (DGGE) to identify serovars of leptospira. The V3 region of most conserved 16S rDNA of five pathogenic leptospiral serovars and one saprophytic serovar was characterized. DGGE method was employed to separate the amplified V3 region based on the nucleotide sequence. On DGGE, amplified V3 region of leptospiral serovars, under study, showed bands at different positions indicating DGGE as the effective method of characterization in the future. DNA sequencing of V3 region of the three serovars showed great difference in nucleotide sequence supporting the results of DGGE.

Disseminated histoplamosis.

A case of disseminated histoplasmosis in a 45-year-old male patient with acquired immunodeficiency syndrome (AIDS) from Pune is reported. The patient presented with high-grade fever and pain in hypochondrium. Clinical signs were pallor and hepatosplenomegaly. Bone marrow and splenic aspirate revealed numerous intracellular oval shaped yeast forms. Histoplasma capsulatum was isolated from the bone marrow and splenic aspirate. H. capsulatum infection is an opportunistic infection usually reported from patient with AIDS in areas endemic for H. capsulatum. The present case highlights the fact that histoplasmosis could be an emerging opportunistic infection in India.

Sero surveillance of leptospirosis among sewer workers in Pune.

Leptospirosis is an important occupational disease affecting people coming in contact with animals and their discharges. The occurrence of infection in ones workplaces is linked to the environment to which the worker is exposed and the adaptability of the organism in that working environment. Rodents usually abound in underground sewers and are carriers of leptospira. The urine of rodents and other animals present in that area is likely to contaminate these sewers. Leptospira are excreted in the urine of infected animals. Thus sewer workers are at a potential risk of leptospirosis. The prevalence of leptospirosis in these workers could thus indirectly predict the presence of the disease in animals in a particular geographical niche. Total seventy-eight sewer workers from 5 different municipal wards in Pune were examined to find out the evidence of past infection with leptospira using microagglutination test (MAT). The prevalence rate was found to be 16.6%. The serovars to which antibodies were detected include autumnalis (38.4%), pyrogenes (23.0%), canicola (15.3%) and pomona (15.3%). Evidence of leptospiral infection was found to be maximum in sewer workers in the areas of the city that were infested with rodents and stray animals.

Neonatal outbreak of salmonella worthington in a general hospital.

This is a report of an outbreak of Salmonella worthington in neonates at Sassoon General Hospitals, Pune, which occurred during August and September 2000. A total of 148 blood, 4 CSF and 6 stool specimens were received from neonates admitted to the hospital. Salmonella worthington could be isolated from 17 clinical specimens [15 blood (10.8%), 1 CSF (25%) and 1 stool (16.6%) sample]. Environmental swabs were also processed. An attempt was made to detect carriers in health care personnel. However, no source or carrier of Salmonella worthington in the hospital environment could be identified. Intensive cleaning measures and fumigation were undertaken. All these measures succeeded in aborting the outbreak.

Evaluation of clinical criteria for the diagnosis of leptospirosis.

OBJECTIVES: Leptospirosis has a wide range of clinical presentation and therefore, clinical suspicion of the infection is often difficult. The objective of this study is to find out the usefulness of the clinical and epidemiological criteria in the diagnosis of leptospirosis and its comparison with microagglutination test (MAT). METHODS: A total of 118 patients with undiagnosed fever of more than seven days duration were included in the study. Their clinical presentation was scored on the basis of a clinical criteria. Sera of the patients were tested for antibodies against leptospira with the help of microagglutination test using a battery of antigens. The usefulness of the criteria was evaluated and compared with microagglutination test. RESULTS: A total of 44 out of 118 (37.28%) patients could be provisionally diagnosed as cases of leptospirosis on the basis of the clinical criteria. Eighteen of these 44 (40.9%) patients had serological evidence of leptospirosis. The criteria had a sensitivity of 81.81%, specificity of 72.91%, a positive predictive value of 40.9% and a negative predictive value of 94.59% when compared with microagglutination test. CONCLUSIONS: The criteria had a moderate sensitivity and specificity. Considering the non-specific signs and symptoms of this infection, the positive predictive value is significantly high. The criteria has a high negative predictive value and this would help the clinicians exclude the diagnosis of leptospirosis with precision.

Common infecting leptospiral serovars in and around Pune, Maharashtra.

Leptospirosis is a disease with protean manifestations. The present study was conducted in Pune to examine the possibility of leptospiral infection among a group of patients with fever of undetermined origin and to identify the common infecting serovars. Serological evidence of leptospirosis was found in 22 of the 118 (18.6%) patients with the help of microagglutination test (MAT) using a battery of 9 antigens. The serovars responsible for infection included autumnalis in eight cases, copenhageni in six, pomona in three, grippotyphosa in two and australis, batavia and canicola in one case each. Thus, there appears to be a focus of leptospirosis in and around Pune with autumnalis and copenhagni as the common infecting serovars.

Contaminated antiseptics--an unnecessary hospital hazard.

A total of 284 antiseptic solutions were studied to check for their sterility. The overall antiseptic contamination rate was 15.14%. 14.85% of freshly prepared antiseptics were contaminated. Here, the problem could be attributed to inadequate precautions while preparing the antiseptics. 15.3% of the in-use antiseptics were contaminated. This could be due to improper handling. Non-fermenters (45.45%), Pseudomonas aeruginosa (30.30%) and Klebsiella spp. (22.72%) were the commonest organisms recovered from the antiseptics. In 44.44% of patients, the isolates obtained from the catheterised urine in the same wards matched with the isolates from antiseptics of that ward. Antiseptic solutions have to be regularly monitored. If they are found to be contaminated, they should be discarded immediately and replaced by fresh sterile antiseptics otherwise instead of preventing infection, antiseptics will become a source of hospital-acquired infection.