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The present study was conducted to correlate the biotypes of Gardnerella vaginalisstrains isolated from cases of bacterial vaginosis and their virulence factors. Thirty-two strains of G. vaginalisisolated from cases of bacterial vaginosis were biotyped. Adherence to vaginal epithelial cells, biofilm production, surface hydrophobicity, phospholipase C and protease activity were tested on these isolates. Biotype 1 was the most prevalent (8; 25%), followed by biotype 2 (7; 21.9%) and biotypes 5 and 8 (5; 15.6%). We did not find any statistical correlation between G. vaginalisbiotypes and its virulence factors. Virulence factors expressed by G. vaginaliswere not associated with a single biotype.

Enterotoxigenic Escherichia coli(ETEC) is an important pathogen responsible for secretory diarrhoea. The production of heat labile enterotoxin (LT), by ETEC, is largely responsible for the pathogenesis of diarrhoea. In the present study we investigated the effect of stress factors such as temperature, pH, osmotic stress and nutritional limitation on the production of LT by ETEC using in-house GMI-ELISA. Four strains of E. coliconsisting, one standard strain MTCC 723 and three clinical isolates were used in the study. Maximum amount of LT (OD 3.285) was produced at 37°C followed by 40°C (OD 3.305). Growth of E. coliin medium with pH 8.6 resulted in maximum amount of LT production (OD 3.489). LT was not detectable when bacteria were grown in medium with pH ≤7.2 and ≥9.2. Sodium chloride concentration of 0.2 M stimulated maximum amount of LT production. Maximum amount of LT was produced when the bacteria were grown in medium containing 2.5g/l of glucose. All the stress factors had a significant effect on the LT production by E. coli, though quantitative differences in the various strains were observed.

Purpose:To eliminate pathogenic bacteria, the host presents conditions that are stressful for bacteria. Oxidative stress arises when the concentration of pro-oxidants like hydrogen peroxide (H2O2) and superoxide anion increases to a level over the basal defence capacity of the cell. In the present study, we studied the effect of oxidative stress on the production of certain virulence factors by Escherichia coli. Methods:E. coliwas exposed to oxidative stress by growing in the presence of different concentrations of H2O2. The effect of oxidative stress on the expression of surface hydrophobicity, adherence, haemolysin production, serum resistance and phagocytosis was studied. Results:Oxidative stress caused a significant decrease in the expression of all the virulence factors of E. coli. Conclusions:Synthesis of virulence factors can be significantly altered by oxidative stress and such changes may affect the pathogenicity of E. coli.

Purpose:To determine the virulence factors produced by Escherichia coliisolated from extraintestinal infections, to study the drug resistance pattern in E. coliwith special reference to extended spectrum β-lactamase (ESBL) and to evaluate screening methods for ESBL. Methods:A total of 152 isolates of E. colifrom various extraintestinal infections were screened for virulence factors such as haemolysin, surface hydrophobicity, serum resistance and protease. All the isolates were also studied for antibiotic susceptibility pattern using modified Kirby Bauer disk diffusion method. ESBL production was screened by standard disk diffusion method and confirmed using phenotypic confirmatory method. Results:Among 152 isolates, 36 (23.7%) were haemolytic, 42 (27.6%) were hydrophobic, 132 (86.8%) were serum resistant and only four were positive for protease. Multiple virulence factor were observed in 67 (44%) of isolates. Seventy-nine (51.4%) isolates produced ESBL. ESBL producing isolates showed multidrug resistance. There was a significant association (P< 0.001) between multiple virulence factors and ESBL production by extraintestinal E. coli. Conclusions:The present study shows the expression of virulence factors and multidrug resistance in E. coliisolated from various extraintestinal infections. The study also shows that appropriate methods of detecting drug resistance and ESBL production are required for the judicious use of antibiotics in managing these infections.

Purpose:The purpose of the present study was to determine the degree of expression of virulence factors such as adherence, cell surface hydrophobicity (CSH) and production of proteinase by different morphological forms of Candida albicanscausing oral candidiasis in human immunodeficiency virus (HIV)-infected individuals. Methods:C. albicans3153A and two strains isolated from oral thrush in HIV infected individuals were induced to undergo phenotypic switching by exposure to UV light and the degree of expression of virulence factors by the different morphological forms was studied. Results:Three different morphological forms of C. albicanswere obtained namely, star (S), wrinkled (W) and ring (R) types from the original smooth (O) variety. It was found that proteinase production was greatest with the W type followed by the R type and O type. The S type produced the least proteinase. Expression of cell surface hydrophobicity and adherence was greatest in the O type followed by the R and then the W type and finally the S type. Conclusions:The differential expression of virulence factors occurs with different phenotypic forms of C. albicansand this may provide a particular morphological type with a distinct advantage over other types in causing candidiasis.

Purpose:To determine the role of humoral immune response and bacterial adherence in the pathogenesis of symptomatic and asymptomatic urinary tract infection in women. Methods:The study population consisted of 30 women with symptomatic UTI, 30 women with asymptomatic UTI and 30 healthy women as controls. Bacterial adherence to vaginal epithelial cells was studied and the concentration of serum and urine antibodies to mixed coliform antigen and clinical isolate was determined by ELISA. Surface hydrophobicity of the urine isolates was determined. Student’s unpaired t test and Pearson’s correlation coefficient test were used in the statistical analysis. Results:Compared to asymptomatic UTI, significantly more number of bacteria adhered to the epithelial cells of women with symptomatic UTI (P<0.001). All cases of UTI had significantly high concentration of urinary IgG antibody to mixed coliform antigens. Asymptomatic UTI cases had higher concentrations of urinary IgG, IgM and IgA antibodies to clinical isolate. Concentration of sIgA level was more in symptomatic UTI. Significant correlation was observed between urinary IgG and adherence of clinical isolate in case of asymptomatic UTI. Conclusions:The present study showed that greater receptivity of epithelial cells to bacteria may increase the susceptibility to UTI. Humoral immune response and local immunity may modify the pathogenesis of UTI.

The study was conducted to determine the antibiotic susceptibility profile of community-associated methicillin resistant Staphylococcus aureus(CAMRSA) strains isolated from infections. S. aureusstrains were isolated from clinical specimens using the standard procedures. CDC definition was used to classify CAMRSA. Antibiotic susceptibility test was done using Kirby-Bauer disk diffusion method. Double disk diffusion method (D-test) was used to detect inducible macrolide, lincosamide and streptograminBresistance (inducible MLSBresistance). A total of 83 CAMRSA were isolated from abscesses and other skin infections in persons without known risk factors for MRSA infection. All CAMRSA were susceptible to vancomycin. Out of 83 CAMRSA, 13 (15.65%) were D-test positive (inducible MLSBpositive) and 6 (7.23%) were positive for constitutive MLSBresistance. Eight strains (9.63%) were resistant to tetracycline and 26 (31.32%) strains were resistant to erythromycin. Increased rate of inducible clindamycin resistance among CAMRSA indicates the importance of identification of such strains by D test to avoid treatment failure when clindamycin is used.

Human immunodeficiency virus-infected patients attending skin outpatient department were studied for nasal carriage of methicillin-resistant Staphylococcus aureus(MRSA) and associated factors affecting nasal colonization. Nasal swabs were used for isolation of S. aureus. MRSA were detected by agar screen and agar dilution methods. Careful examination for dermatoses was carried out. Forty-six of the 60 (76.67%) outpatients with HIV infection were colonized with S. aureusin the anterior nares. Significant number of S. aureuscarriers were in the 31–40 year age group. Methicillin resistance was found in eight (17.39%) isolates. Of the 46 S. aureusstrains, 29 (63%) were resistant to erythromycin, 69.5% to co-trimoxazole and 41.3% to ciprofloxacin. Co-trimoxazole use was found to be a risk factor for S. aureuscarriage (P= 0.0214) but not for methicillin resistance. Hospital stay for more than 10 days was a risk factor for methicillin resistance whereas stay for more than 25 days was found to be a highly significant risk factor. Dermatophytosis and herpes simplex virus infection were other risk factors for nasal carriage of S. aureus.

Sixty intact, non-carious and unrestored teeth extracted due to periodontal disease were used to determine the most effective method of sterilisation. The teeth were divided into six groups, each containing 10 teeth. Group 1 teeth were immersed in 10% formalin for seven days, group 2 teeth were immersed in 3% hydrogen peroxide for seven days, group 3 teeth were immersed in 2.6% sodium hypochlorite for seven days, group 4 teeth were boiled in water at 100°C for 20 minutes, group 5 teeth were autoclaved at 121°C at 15 lbs psi for 30 minutes, and group 6 teeth were immersed in normal saline for seven days. After the treatment, the teeth were individually inoculated into trypticase soy broth and incubated for 48 hours. A questionnaire survey was also conducted to determine the awareness of dental students regarding infection due to extracted human teeth and the common disinfection/sterilisation methods used. Autoclaving at 121°C, 15 lbs psi for 30 minutes and immersion in 10% formalin for seven days were effective in disinfecting/sterilising extracted human teeth. Chemicals such as 2.6% sodium hypochlorite, 3% hydrogen peroxide and boiling in water were not effective. The results indicate that autoclaving for 30 minutes or immersion in 10% formalin for seven days could be effectively used for disinfection/sterilisation of extracted human teeth.