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We evaluated the safety and potency of the Kyasanur Forest disease (KFD) vaccine inactivated with different formalin concentrations in mice, since the side effects due to higher formalin concentrations have been a major reason for vaccine refusal. Furthermore, with an objective to reduce the use of mice in vaccine testing, we performed quantification of the KFD virus by real-time PCR and compared it with in vivo titration in mice. The KFD vaccine prepared in chicken embryo fibroblast cells was inactivated with 0.04%, 0.06%, and 0.08% concentrations of formalin. The vaccine inactivated with 0.04% and 0.06% formalin failed the safety test, whereas the KFD vaccine inactivated with 0.08% formalin was safe and potent with a log protective index of 5678 in mice. This reduced formalin content may induce no/lesser side-effects of pain/swelling which may increase the vaccine acceptance. The real-time PCR on individual KFD vaccine harvests interpreted that when the CT value of each harvest is

Aim: To estimate antibody titres to Hepatitis B surface antigen (HBsAg) among vaccinated medical students. Study Design: Cross sectional study Place and Duration of Study: Study was done in the Department of Microbiology, Adichunchanagiri Institute of Medical Sciences, B.G. Nagara between October 2019 to November 2019. Methodology: Study included 281 medical students who had taken three doses of Hepatitis B vaccination. Informed consent and Institutional Ethical clearance was obtained. Under aseptic precautions, 5 ml of venous blood was collected and serum was separated by centrifugation at 3000 rpm for 5 min. Serum were stored at -200 C. Antibodies to Hepatitis B surface antigen were estimated by Electro Chemiluminescence method (Anti-HBs G2 Elecsys cobas e 100- Roche Diagnostics India Pvt. Ltd.) as per manufacturer’s instruction using quality controls. Results: In the present study, 98.9% of students had Anti HBs antibody titres >10mIU/ml. Only one (1.1%) student had titre 1000 mIU/ml and 15 (16.7%) had antibody levels >100≤1000 mIU/ml. Higher seroconversion rates were noted among female students (100%) but was not statistically significant (P value=0.34) Conclusion: Except for one, all other study participants were found to have protective antibody titres. Vaccination cards containing the vaccination details and the antibody titres should be made available to all medical students to guide infection control practices in case of accidental exposure.

Methicillin resistant Staphylococcus aureus (MRSA) has emerged as one of the commonest causes of hospital acquired infections. Vancomycin is the drug of choice for the treatment of MRSA. Increase in Minimum inhibitory concentration (MIC) of Vancomycin has been observed in both Methicillin sensitive and resistant Staphylococcal isolates. In critically ill patients, Vancomycin has become a poor therapeutic antibiotic whereas; Linezolid has emerged as an alternative drug in treating such patients. This study was undertaken to determine the sensitivity pattern among clinical isolates of Staphylococcus aureus (S. aureus) to Vancomycin and Linezolid, in order to formulate a better treatment.100 S. aureuswere isolated from various clinical specimens. Antibiotic sensitivity testing was performed by Kirby Bauer disc diffusion method and MICs of Vancomycin and Linezolid were determined by E-test following CLSI guidelines. Out of 100 S. aureus isolated, 68 were MRSA strains. Among 68 strains of MRSA for which MIC levels of Vancomycin 4 µg/ml, 2µg/ml and 1 µg/ml were 4, 27, and 37 respectively. Similarly, out of total MRSA strains MIC levels of Linezolid 4 µg/ml, 2µg/ml and 1 µg/ml were 1, 25, and 42 respectively. All 100 strains showed similar in-vitro efficacy for Vancomycin and Linezolid by Kirby Bauer disc diffusion method, but the number of strains with higher ranges of MICs of Vancomycin were more as compared to those which had higher ranges of MICs for Linezolid. Hence we suggest that Linezolid can be used as an alternative for the treatment of MRSA.

Introduction: Infections due to Candida especially Non-albicans Candida (NAC) species are increasingly being reported in recent years. Isolation and identification up to species level are of utmost importance in the early management of these infections. Materials and methods: All the clinical samples received in the Microbiology laboratory were inoculated on to Blood agar and MacConkey agar. The isolates which revealed Gram-positive budding yeast cells on Gram staining were further inoculated on to SDA (Sabouraud Dextrose agar) and Hicrome Candida differential agar. Candida was differentiated into species based on color and morphology on Hicrome agar. Result: A total of 126 Candida were isolated from various clinical samples. C.Albicans was the most common species isolated (49.2%) followed by C. tropicalis (19.8%), C. glabrata (19%), and C. krusei (12%). Conclusion: Hicrome agar is a simple, rapid, and cost-effective medium for the speciation of Candida.

Lumpy skin Disease (LSD) is a vector borne pox disease of domestic cattle and Asian water buffaloes that is characterized by the appearance of skin nodules. The incubation period varies between four to 14 days. It may go up to five weeks under field conditions. The clinical signs include nasal and ocular discharge, fever, reduction in milk production and characteristic skin lesions. Initially the sub scapular and prefemoral lymph nodes get enlarged, followed by high fever of 105 °F which may last for a week. Generally fever precedes the appearance of skin lesions and may be accompanied by a sharp drop in milk production. Lacrimation and nasal discharge are generally seen before onset of clinical signs. Nasal discharge becomes mucopurulent as disease progresses, mild clinical cases show few nodules on the body whereas in severe cases the entire body will be covered with skin lesions. Lesions may also be seen in the digestive and respiratory tract and also on almost all the internal organs. Pneumonia is a common finding due to secondary bacterial infection. Lesions can also be seen in the mucus membrane of the eyes with mucopurulent discharge. Painful ulcerative lesions may also be seen in the cornea leading to blindness.

Background: Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the most common cause of chronic liver disease (CLD) worldwide. Coinfection with both HBV and HCV can occur because of shared routes of infection. Therefore, this study was performed to investigate the seroprevalence of HBV and HCV dual infection among patients attending K R hospital, Mysore with underlying CLD manifestations. Materials & Methods: Serum samples from 80 clinically diagnosed chronic liver disease patients attending K R hospital, Mysore was screened for the presence of hepatitis B surface antigen, Anti-HBc IgM, Anti-HBc Ig Gand anti-hepatitis C virus antibodies by ELISA. Serum samples from 15 healthy individuals were also screened. Results: Among 80 chronic liver disease patients, 67 (83.75%) were males and 13 (16.25%) were females. The maximum number of CLD patients was in the age group of 41 – 50 years (41.30%). Alcoholism (72.5%) was the most common risk factor & cirrhosis of liver was the most common clinical presentation (81.25%). 38 (47.5%) cases were HBV positive (positive for any one marker for HBV). HBs Ag was positive in 22 cases, anti-HBc Ig Min 7 cases, anti-HBc Ig Gin 29 cases. 4 (5%) cases were positive for anti HCV. Three (3.75%) cases showed coinfection of HBV & HCV. Conclusion: To prevent the spread of HBV and HCV, people must be educated about these infections and their mode of transmission. All CLD patients should be tested for HBV and HCV to prevent the mortality and morbidity. HBV & HCV coinfection should not be excluded by negative HBsAg status alone.

measure to the persistence of infections as a major cause of morbidity and mortality.The purpose of this study was to catalog the carrier prevalence of MDR bacteria at the community level. It was mainly focused on isolating carriers from anterior nares and the gut and to determine their antimicrobial susceptibility pattern at admission. A total of 100 patients were screened for carriage prevalence of MDR bacteria in anterior nares and gut . Of the 158 isolates obtained from the stool samples, 48 (30.3%) were MDR, 34 (21.5%) were XDR and 3 (1.8%) were PDR. One more point worth noting was the pan-drug resistant organisms in the gut of 3 (03%) patients. 2 (66.6%) of these isolates were E.coli and the other was Enterococci. On the contrary, of 83 isolates obtained from nasal samples 18 (21.6%) were MDR and 7(8.43%) were XDR. (26.4%) of the Staphylococci obtained were multi-drug resistant. 20(29.4%) of the Staphylococci were MRSA. Of these 14 (70%) could be CA-MRSA. 33 (24.6%) and 30(22.3%) of the GNBs obtained from the stool samples were ESBL and MBL producers respectively.Thirteen (54.1%) of the isolates of Enterococci were multi-drug resistant. 10 (41.6%) of the Enterococci were resistant to vancomycin. In this situation, it is perhaps better for all the health care institutions to come up with their own screening strategy, in order to the curb the evolution of these multi-drug resistant bacteria to a destructive rate.

BACKGROUND: Extra pulmonary tuberculosis (EPTB) is a major health problem due to its destructive nature. The term EPTB, which encompasses all forms of TB other than pulmonary TB (PTB), has a broad spectrum of clinical manifestations, hence a real diagnostic challenge. AIM: The primary objective of this study was to describe the basic demographic, clinical characteristics, trends of various types of tuberculosis and to detect the incidence of EPTB in Mysore over a period of two years. MATERIALS AND METHODS: This is an observational study of patients of all types of Extrapulmonary tuberculosis from January 2011 to December 2013 in all age groups, evaluated at Mysore Medical College and Research institute, Mysore, Karnataka, India. RESULTS: Among 4500 cases registered for treatment of all forms of tuberculosis, 710(15.77%) had EPTB. More than half of the cases of EPTB (n = 472, 66%) were among adult age groups. Pleural TB was the commonest type of EPTB (n = 268, 37.03%), followed by lymph node TB (n = 210, 29.81%). Involvement of lymph nodes was the commonest manifestation in less than 14 years of age group, while involvement of pleura was more common among > 65 years. CONCLUSION: The burden of EPTB is more among the productive age group. Increase in the trend of EPTB cases, and other rare forms of EPTB are a point of concern highlighting the importance of strengthening the services towards this group.

Background: Candida species are known to be the most common fungal pathogens isolated from blood cultures of neonates. Recent reports from our country indicates trend towards an increasing prevalence of non-albicans candidemia. Candida species possess a number of virulence factors which enable them to cause hematogenously disseminated infections in susceptible hosts with increased morbidity and mortality. Objectives:To know the prevalence of candidemia in neonates and their antifungal susceptibility pattern. Materials and Methods: Blood samples from suspected cases of neonatal septicemia were subjected to culture, incubated for 7 days and subcultures performed. Culture yielding pure growth of Candida were included for the study and identified by standard methodology. Antifungal susceptibility was performed towards fluconazole and voriconazole by disc diffusion method on Muller-Hinton agar with 2% glucose and methylene blue. Result: Among 1583 blood cultures 80(5%) candida isolates were obtained. The various species isolated were C.pseudotropicalis 34(42.5%), C.tropicalis 22(27.5%), C.albicans 12 (15%) C.guillermondii 6(7.5%), C.parapsilosis 4(5%) and C.krusei 2(2.5%). Fluconazole susceptibility was observed in 56(70%) sensitive, 18 (22.5%) Susceptible-Dose Dependent (SDD) and 6(7.5%) resistant and all were sensitive to voriconazole. Conclusion: In this study non albicans candida was the common isolate & they showed decreased resistance to Fluconazole. In neonatal septicemia speciation & antifungal susceptibility may help in management.

Background: There is a need to generate data from India on relative frequencies of specific opportunistic infections (OIs) in different regions and their relation to the choice of commonly used generic highly active anti-retroviral therapy (HAART) regimens. Objectives: To document the prevailing prevalence pattern of OIs both before and after HAART, to look for reduction in OIs following HAART, to assess the risk of developing new OIs within 6 months of HAART initiation and to see if there is any difference in the risk of developing a new OI within 6 months of HAART initiation, for those on Efavirenz (EFV)-based regimens and Nevirapine (NVP)-based regimens. Materials and Methods: In a prospective observational cohort study conducted in South India involving 108 ART-naive AIDS patients, different pathogens were isolated and identified using standard laboratory techniques. Data analysis was done using SPSS software (version 16.0). Risk of developing an OI after HAART initiation was assessed using the likelihood ratio test from Cox regression models. Results: Tuberculosis (53.4%), oral Candidiasis (27.2%) and Herpes Zoster (14.7%) were the common infections seen. There was a drastic reduction of 96.59% in OI events after 6 months of HAART. The risk of developing an OI within 6 months of HAART initiation was 5.56%. Time to development of an OI in the first 6 months of HAART was shorter for the NVP-based regimens than with EFV-based regimens, but this difference was not statistically significant (HR=0.891, 95% CI: 0.179-4.429; P=0.888). Conclusion: Tuberculosis is the most important OI before initiation of HAART. Both EFV and NVP-based regimens are equally efficacious in controlling OIs.

Although bovine viral diarrhoea virus (BVDV) is prevalent in Indian cattle causing economic losses in cattle farming, its detection in bull semen has not yet been reported. Following passage of raw bull semen (n = 4) on MDBK cells, testing for BVDV was done by antigen ELISA and real-time RT-PCR. BVDV type-2 (BVDV-2) was identified in three samples from southern India by real-time RT-PCR. Genetic typing of the 5'-UTR sequences classified all the three BVDV strains as BVDV-2a subtype. These were found genetically closely related to the strains from USA, but divergent from the BVDV-2a strains from northern India. Phylogenetic analysis of N pro sequences confirmed the findings. The results provide evidence of circulation of BVDV-2a strains in southern India. The detection of BVDV in bull semen from India highlights the importance of mandatory testing of breeding bulls and bull semen for BVDV to minimize the risk of BVDV transmission.

INTRODUCTION: Mupirocin is mainly used for decolonisation of Methicillin-resistant Staphylococcus aureus among patients and health-care workers. However resistance originates from coagulase negative staphylococci (CoNS) and spreads to S. aureus by horizontal gene transfer. Hence, the present study was done to determine the overall prevalence of high- and low-level Mupirocin resistance among S. aureus and CoNS and correlate with the occurrence of Methicillin and inducible Clindamycin resistance. MATERIALS AND METHODS: A total of 100 consecutive, non-repetitive, clinical isolates of Staphylococci were obtained from various samples in a rural tertiary care hospital of Karnataka between August and September 2016. Antibiotic susceptibility testing was done according to the Clinical and Laboratory Standards Institute (M100-S25, 2015) guidelines. Low- and high-level Mupirocin resistance was screened using 5 μg and 200 μg discs, respectively. RESULTS: Of 100 staphylococcal isolates, high-level Mupirocin resistance (MupRH) was detected in 13 isolates and low-level Mupirocin resistance (MupRL) was found in 4 isolates. Of 13 HLMR isolates, majority were CoNS (12 out of 13, 92.31%). Even in LLMR isolates, 75% (3 out of 4) were CoNS. All the HLMR and LLMR isolates were Methicillin resistant (P < 0.05). CONCLUSION: High prevalence of Mupirocin resistance was found in the present study. Hence, screening for Mupirocin resistance should be routinely done and the drug must be used judiciously.

Bacillus cereus is ubiquitous in nature and while most isolates appear to be harmless, some are associated with food-borne illnesses, wound infections, endocarditis, osteomyelitis, endophthalmitis and urinary tract infections in humans. Recently, a few isolates have been identified as the causative agents of anthrax-like severe pneumonia in humans, and these isolates were found to harbor most of the B. anthracis virulence plasmid pXO1. Here we report the characterization of three clinical B. cereus isolates recovered from heart blood and spleen samples of cattle which had died with ‘anthrax like’ symptoms. Apart from the cultural characterizations, primers targeting the 16S rRNA gene of B. cereus were designed and used on these isolates. The isolates were found to harbor the pXO1 plasmid and lacked pXO2 plasmid. Further characterization of the pXO1 plasmid revealed that the isolates contained pag, lef and cya genes, which code for protective antigen, lethal factor and edema factor toxins responsible for eliciting an ‘anthrax like disease’ in cattle. The sequencing and phylogenetic analysis of partial pag gene sequences of B. cereus isolates were identical to pag gene sequences on the pXO1 of B. anthracis. In a pathogenicity test on mice, B. cereus isolates, when inoculated by the intra peritoneal route, caused mortality of the mice within 6 hours post inoculation., Bacillus cereus je posvudašnja bakterija. Većina izolata te bakterije je neškodljiva, a neki mogu uzrokovati bolesti koje se prenose hranom, infekcije rana, endokarditis, osteomijelitis, endoftalmitis i infekcije mokraćnog sustava u ljudi. Nedavno je identificirano nekoliko izolata koji uzrokuju tešku upalu pluća u čovjeka sličnu onoj kod bedrenice. Ti izolati većinom nose virulentni plazmid pXO1 vrste B. anthracis. U ovom radu određena su obilježja triju kliničkih izolata vrste B. cereus izdvojenih iz krvi sadržane u srcu i uzoraka slezene goveda uginulih pod znakovima sličnima bedrenici. Osim određivanja kulturalnih obilježja, pripremljene su i početnice za gen 16S rRNA vrste B. cereus koje su bile rabljene za identifikaciju izolata. Ustanovljeno je da izolati nose plazmid pXO1, a nedostaje im plazmid pXO2. Daljnja karakterizacija plazmida pXO1 pokazala je da izolati sadrže gene pag, lef i cya koji kodiraju za zaštitni antigen, letalni čimbenik i edemski čimbenik, toksine koji su odgovorni za pojavu bolesti u goveda slične bedrenici. Određivanje slijeda i filogenetska analiza dijela sekvencija gena pag izolata B. cereus pokazala je da su oni istovjetni sekvencijama gena pag na pXO1 bakterije B. anthracis. U testu patogenosti na miševima, izolati B. cereus prouzročili su njihovo uginuće šest sati nakon intraperitonejske inokulacije.

A retrospective study on the epidemiology of foot and- mouth disease (FMD) in Karnataka, India between the years 1977 and 2012-13 based on the data collected through passive and active surveillance was undertaken. A total of 11,159 outbreaks with 0.271 million cases of FMD were recorded from 30 different revenue districts of Karnataka. There was a significant difference between the years for the annual incidence of FMD (P =

Background & objectives: Owing to the ever-expanding access to HAART (highly active anti-retroviral therapy) in resource-limited settings, there is a need to evaluate alternate markers like absolute lymphocyte count (ALC) as a surrogate for CD4 counts. This study was done to assess the usefulness of ALC as a surrogate marker for CD4 counts in monitoring HIV-infected patients after HAART initiation. Methods: In this study, 108 HIV-positive adult patients of both sexes fulfilling the inclusion criteria were included. CD4 and ALC were recorded at baseline. After initiation on HAART, these patients were followed up at three month intervals. Results: ALC and CD4 counts were positively correlated (Spearman correlation coefficient= 0.553). After six months of HAART, the sensitivity of an ALC increase as a marker for CD4 count increase at six months was 82 per cent, specificity was 100 per cent, PPV was 100 per cent and NPV was 31 per cent. Area under the corresponding ROC curve for CD4 increase of >100 cells/μl was 0. 825 ± 0.053. Interpretation & conclusions: ALC may be a useful surrogate marker in predicting an increase in CD4 counts as a response to HAART, but of questionable value in predicting a decrease in CD4 counts.

Methods The study group included 106 HIV positive patients with history of heterosexual contact and the control group included 30 asymptomatic healthy individuals. The sera of patients were tested for HBsAg and antiHCV using third generation ELISA kits. The samples positive for first kit were retested for confirmation of results. The sera of control group were simultaneously screened for HIV, HBsAg, and anti-HCV by ELISA method.

Introduction: In resource-limited settings, due to the high cost of CD4 cell count testing, physicians must decide about opportunistic infection (OI) prophylaxis without a laboratory evaluation of HIV stage and level of immune suppression. This study aimed to evaluate the correlation of total lymphocyte count (TLC), an inexpensive laboratory parameter, to CD4 count, and to determine a range of TLC cut-offs for the initiation of OI prophylaxis that is appropriate for resource-limited settings. Methodology: Spearman correlation between CD4 count and TLC was assessed in patients attending the Anti-Retroviral Therapy (ART) centre at Mysore, India. Positive predictive value (PPV), negative predictive value (NPV), sensitivity, and specificity of various TLC cut-offs were computed for CD4 counts < 200 cells/mm3. Correlation and statistical indices were computed for all patients and for HIV patients with active tuberculosis. Results: Good correlation was noted between the 106 paired TLC and CD4 counts (r = 0.3497).TLC < 1200cells/mm3 had 88.14% sensitivity and 34.78% specificity for CD4 count < 200 cells/mm3. In those patients with active tuberculosis, TLC< 2000cells/mm3 had 95.24% sensitivity and 100% specificity for CD4 count < 200cells/ mm3. Conclusions: TLC could serve as a low-cost tool for determining when to initiate prophylaxis in resource-constrained settings.

DNA samples from 19 reference serovars belonging to 19 different serogroups of Leptospira interrogans and two serovars belonging to Leptospira biflexa were examined by bacterial restriction endonuclease analysis using EcoR I and Hae III enzymes. All the serovars gave unique restriction patterns that differed from each other. DNA from 10 local isolates digested with these enzymes produced patterns which on comparison with the standard patterns produced by reference strains could be identified to serovar level.

Background: Cardiovascular disease, resulting from atherosclerosis, is a leading cause of global morbidity and mortality. Classical risk factors explain much of the attributable risk for cardiovascular events, but other risk factors for the development and progression of atherosclerosis, which can be identified, may be important therapeutic targets. Infectious agents, such as Chlamydia pneumoniae, have been proposed as contributory factors in the pathogenesis of atherosclerosis. The present study was conducted to determine the seroprevalence of C. pneumoniae antibodies and to study the association of chronic C. pneumoniae infection with Coronary Artery Disease (CAD). Methods: The study group included 90 angiographically proven CAD patients and age and sex matched 90 normal coronaries as control group. With total aseptic precaution 3 ml blood was collected. Enzyme linked immunosorbant assay was performed for all subjects to detect the presence of IgG and IgA antibodies to Chlamydia pneumoniae (Cp). Results: IgG and IgA Cp antibodies were detected in 67.8% and 58.9% CAD patients compared to 45.6% and 11.1% controls. IgG + IgA Cp antibodies were detected in 88.9% CAD patients when compared to 50.0% controls. Seroprevalence of IgG and IgA Cp antibodies were high among CAD patients compared to controls and was found statistically significant. A significant presence of Chlamydia pneumoniae antibodies was detected in smokers, diabetes mellitus, hypertension, and dyslipidemia. Conclusion: In the present study, the seroprevalence of IgG and IgA Cp antibodies was found to be higher in CAD patients compared to controls. The present study supports the association between Chlamydia pneumoniae infection and Coronary artery disease.

Background: The global impact of Tuberculosis (TB) and Human Immunodeficiency Virus (HIV) co-infection is one of the major public health challenge. India has a very high burden of TB according to the WHO. A decrease in CD4 counts in HIV-TB co-infection leads to an increase in morbidity and mortality. Methods: Information regarding the duration of HIV, type of TB, CD4 counts before and after ATT and any associated Opportunistic Infections (OIs) were collected from the records of 100 patients with HIV-TB co-infection who attended ART centre for a period of one year. The collected data was statistically analyzed. Results: In the study group, 35 had Pulmonary Tuberculosis (PTB) and 65 had Extra Pulmonary Tuberculosis (EPTB), 40 had OIs. Mean CD4 count prior to ATT in PTB was 197 (7-940), EPTB 192 (13-683) and with OIs 129 (7-288). After completion in PTB was 300, EPTB 302 and 252 in OIs. Least CD4 count of 121 was observed in patients above 50yrs and after completion it was 133. Incidence of both EPTB and PTB was higher in males 66.2% and 62.9%, and in the age group of 31-50 yrs 50.8% and 60% (Cell counts expressed in cells/µl.). Conclusion: In our study, we found that there was significant recovery of CD4 cells following ATT. Difference in CD4 counts among patients with PTB and EPTB was not significant. There was remarkable reduction of CD4 counts in patients who had other OIs and the recovery after ATT was also marginal.