Antibiotic resistance pattern and plasmid profiling of thermotolerant Escherichia coli isolates in drinking water.

BACKGROUND: Antibiotic resistant Escherichia coli is potential source of transmission of resistance to other water borne pathogens where plasmid borne resistance is most significant. METHODS: Drinking water samples were collected from different water sources: that is to say- tap, well and spring from different places of Kathmandu where E. coli and thermotolerant E. coli were isolated using membrane filtration technique. Antibiotic susceptibility was determined using a modified Kirby Bauer disc diffusion method and thermotolerant E. coli isolates from tap water were subjected for plasmid profiling. RESULTS: Type of water sources were not associated with the presence of coliform (P=0.155) and thermotolerant coliform (P=0.235) but the significant association was observed in thermotolerant coliform and thermotolerant E. coli for all sources tap (P=0.029), well (P=0.028), spring (P=0.05) but total coliform and E. coli association was found for well (P=0.01). All E. coli and thermotolerant E. coli isolates were susceptible to Ofloxacin, Chloramphenicol and Cotrimixazole. Resistance to Cefexime, Amikacin, Nalidixic acid, Amoxicillin, Tetracycline were 17 (54.8%), 9 (29%), 11 (35.5%), 25 (80.6%), 29 (93.5%) and 19 (57.6%), 12 (36.4%), 13 (39.4%), 31 (94%), 33 (100%) was observed in E. coli and thermotolerant E. coli respectively where 25 (75.8%) thermotolerant E. coli and 22 (70.9%) E. coli were observed with multiple drug resistance patterns. Single band of plasmid were observed in three MDRs and one non-MDR isolates and size varied from 2kb to >10kb. All Nalidixic acid resistant thermotolerant E. coli were found to harbor a plasmid. CONCLUSIONS: Presence of plasmid in Nalidixic acid resistant thermotolerant E. coli heightens public health issue and the need of monitoring Quinolone resistance bacteria in environment.

The burden of dengue infection in some vulnerable regions of Nepal.

Dengue is an emerging mosquito borne disease of public health importance in Nepal. A descriptive cross sectional study was carried out to estimate sero-prevalence and distribution pattern of dengue in certain vulnerable regions of Nepal from June to September 2009. A total of 460 venous blood samples were collected from individuals experiencing a febrile illness clinically consistent with dengue infection visiting nearby hospitals of Kanchanpur, Kailali, Banke, Dang and Chitwan districts. The sero-prevalence of dengue virus specific IgM was determined by enzyme linked immunosorbent assay (ELISA) kit. The anti-dengue IgM positivity was found to be 12.17%. The higher frequency of positive cases (16.4%) were from age group 20-40 years followed by < 20 years age group with 9.7% and 5.3% from > 40 years age group. The association between dengue infection and age is found to be statistically significant (p < 0.05). The male:female ratio was determined as 1.3:1 in IgM positive population. Among sampling areas, Kanchanpur showed highest prevalence of dengue infection (15.5%) followed by Chitwan (11.7%), Kailai (11.1%), Banke (10.7%) and Dang (8.3%). Similarly, 94.6% of the positive cases were indigeneous and had no history of travel to other countries. Dengue is firmly established in terai region with increasing trends of infection and expansion into newer areas raising a public health threat. Regular epidemiological studies are suggested which could further reveal the contributing factors associated with dengue virus infection and help in formulating strategies in reducing the transmission rate and control of the infection.

Preliminary observations on lymphocyte subpopulations in HIV seropositive & HIV seronegative tuberculosis patients in Pune, India.

CD4 and CD8 lymphocyte counts were determined in 59 HIV seropositive and 41 HIV seronegative newly diagnosed tuberculosis patients in Pune. There were significant differences in the CD4 counts and CD4/CD8 ratios between HIV seropositive and HIV seronegative tuberculosis patients. Majority of the HIV seropositive patients had a CD4 count less than 500 cells/cu.mm, whereas among the HIV seronegative patients, majority had a CD4 count more than 500 cells/cu.mm. In HIV seropositive patients with extrapulmonary and pulmonary tuberculosis, the CD4 counts were lower than in those who had only pulmonary or extrapulmonary tuberculosis. There was no significant differences in the CD8 counts between HIV seropositive and HIV seronegative tuberculosis patients, except for patients with pulmonary cavity, where the CD8 counts were significantly higher in HIV seropositive tuberculosis patients. In HIV seropositive individuals with pulmonary tuberculosis, the CD8 counts in those with pulmonary cavity were higher than in those without any pulmonary cavity. Absence of cavitation and presence of pulmonary with extrapulmonary tuberculosis occurred when immune activation was at a lower level.

Isolation & preliminary characterization of two HIV-2 strains from Pune, India.

Two HIV-2 strains were isolated from peripheral blood mononuclear cells of two HIV-2 seropositive patients with pulmonary tuberculosis by co-cultivating the cells with phytohaemagglutinin-P stimulated heterologous normal lymphocytes. Biological characterization of the isolates indicated that both isolates were syncytium inducing and induced cytopathic effect in the form of giant cells and syncytia formation in four T lymphoid cell lines. The isolates differed in their replication pattern. The isolates were confirmed as HIV-2 by nested PCR using HIV-1 and HIV-2 specific oligonucleotide primers from the env region and by supplementary tests like indirect immunofluorescence assay, syncytium inhibition assay using reference and HIV-2 reactive patients' sera, western blot and electron microscopy. Neutralization of one isolate (TB1) with two Senegal reference sera also indicated that the isolate may be related to the Senegal strain. To our knowledge, this is the first report of isolation of HIV-2 in India.

Increasing trend of HIV seroprevalence among pulmonary tuberculosis patients in Pune, India.

A total of 4618 tuberculosis patients attending the TB clinic at the Sassoon General Hospitals, Pune between 1991 and 1996 were screened for anti-HIV antibodies. Of these 694 were found reactive in enzyme immuno assay (EIA) and 624 were further confirmed by a second test, either rapid EIA or Western blot. HIV-1 reactivity was predominant among tuberculosis patients with HIV-2 reactivity appearing only in 1995. HIV-2 seroreactivity accounted for 0.54 and 1.02 per cent of all HIV reactive samples in 1995 and 1996. HIV-1 and HIV-2 dual reactivity accounted for 1.63 and 2.04 per cent of all infections in 1995 and 1996. The overall seroprevalence of HIV among newly diagnosed tuberculosis patients rose from 3.2 per cent in 1991 to 20.1 per cent in 1996.

PIP: A total of 4618 tuberculosis (TB) patients attending the TB clinic at the Sassoon General Hospitals, Pune, India, between 1991 and 1996 were screened for anti-HIV antibodies. Of these, 694 were found reactive in enzyme immunoassay (EIA) and 624 were further confirmed by a second test, either rapid EIA or Western blot. HIV-1 reactivity was predominant among tuberculosis patients, with HIV-2 reactivity appearing only in 1995. HIV-2 seroreactivity accounted for 0.54% and 1.02% of all HIV reactive samples in 1995 and 1996. HIV-1 and HIV-2 dual reactivity accounted for 1.63% and 2.04% of all infections in 1995 and 1996. The overall seroprevalence of HIV among newly diagnosed tuberculosis patients rose from 3.2% in 1991 to 20.1% in 1996.

Efficacy of stimuliv, an indigenous compound formulation, against hepatotoxicity of antitubercular drugs--a double blind study.

Anti-tubercular drugs are known to cause hepatotoxicity, which may lead to noncompliance to drug therapy. Stimuliv, an indigenous compound formulation, is reported to be useful in liver disorders. Efficacy of prophylactic administration of stimuliv against anti-tubercular drugs-induced hepatotoxicity was studied in this double blind randomized clinical trial. One hundred and forty-five newly diagnosed patients of tuberculosis were included in the study. Out of these, sixty three patients were treated with stimuliv (2 tablets thrice daily), sixty received the placebo, while twenty-two dropped out of the study. The patients were assessed clinically and biochemically at two-week intervals over a period of two months. In stimuliv-treated group, the incidence and severity of hepatotoxicity was significantly less (p < 0.05) as compared to placebo-treated group. In addition, patients treated with stimuliv had better appetite and weight gain. Stimuliv treatment may be recommended in newly diagnosed adult patients of tuberculosis.