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In November 2003, an outbreak (41 cases; attack rate-4.3%; no deaths) of severe diarrhoea was reported from a village in Orissa, eastern India. Thirteen of these cases were hospitalized. A matched case-control study was conducted to identify the possible exposure variables. Since all wells were heavily chlorinated immediately after the outbreak, water samples were not tested. The cases were managed symptomatically. Descriptive epidemiology suggested clustering of cases around one public well. Vibrio cholerae El Tor O1, serotype Ogawa was isolated from four of six rectal swabs. The water from the public well was associated with the outbreak (matched odds ratio: 12; 95% confidence interval 1.2-44.1). On the basis of these conclusions, access to the well was barred immediately, and it was protected. This investigation highlighted the broader use of field epidemiology methods to implement public-health actions guided by epidemiologic data to control a cholera epidemic. [ABSTRACT FROM AUTHOR]

Background: Multiple clinical studies have described the benefits of probiotic Saccharomyces boulardii (S. boulardii) CNCM I-745 against diarrhoea, but the real-world evidence supporting its use is lacking., Objective: To evaluate effectiveness of the S. boulardii CNCM I-745 group in a real-world setting., Methods: This was an electronic medical record (EMR)-based, retrospective, multicentre, comparative study in Indian adult patients presenting with diarrhoea managed between January 2020 and January 2022. Data of patients at the baseline visit, with a follow-up visit within 15 days, and who were administered S. boulardii CNCM I-745 (for the test group) or any other treatment modality excluding probiotics (for the control group) were considered. Effectiveness was evaluated on the basis of number of patients who did not complain of diarrhoea at follow-up., Results: Of 30,385 adult patients with diarrhoea, 270 patients prescribed S. boulardii CNCM I-745 were included, while the control group comprised 1457 patients. The baseline median age of the test group was 47 years (range 19-86 years), while it was 44 years (range 19-100 years) for the control group. The majority of patients in both study groups were females (56.7% in the test and 51.5% in the control group). Median duration between visits was 5 days (range 1-15 days) in both study groups. In all, 77.8% patients (95% CI 72.34-82.59) in the test group did not complain of diarrhoea at follow-up, while the proportion was 15.8% (95% CI 13.95-17.76) in the control group (p < 0.05). Odds ratio (OR) for absence of diarrhoea in the S. boulardii CNCM I-745 group versus the control group was 18.7 (95% CI 13.6-25.7, p < 0.05). For subgroups on concomitant antibiotics, a significant advantage was noted again for the test versus the control group (76.8% versus 18.4%; p < 0.05; OR: 14.7 with 95% CI 8.8-24.4; p < 0.05)., Conclusion: The effect of S. boulardii CNCM I-745 probiotic in controlling diarrhoea was better than anti-diarrhoeal and/or oral rehydration therapy in real-world clinical practice. The effect was similar even with concomitant antibiotic usage., (© 2024. The Author(s).)

Background: The present study reports on the comprehensive analysis of Vibrio cholerae O1 and non-O1/non-O139 serogroups isolated from environmental water sources during cholera outbreaks, epidemics and surveillance studies between years 2007 to 2019 from different districts of Odisha, India., Methods: A total of 85 stocked cultures of V. cholerae O1 and non-O1/non-O139 strains were analyzed for different ctxB genotypes, toxic genes, antibiogram profiles through PCR assays and pulsotyped by pulsed-field gel electrophoresis (PFGE)., Results: From all V. cholerae strains tested, 51 isolates were O1 Ogawa and the rest 34 strains were non-O1/non-O139. All the V. cholerae O1 strains were altered El Tor variants carrying ctxB1, ctxB3 and ctxB7 genotypes. However, only ctxB1 genotypes were present in V. cholerae non-O1/non-O139. Though non-O1/non-O139 strains were negative by O1 antisera, 20% strains were positive for rfbO1 gene by PCR assay. All the V. cholerae isolates possessed a variety of virulence genes including ace, ctxAB, toxR, zot, hlyA which were in higher percentage in the case of V. cholerae O1. The Vibrio cholerae O1 and non-O1-/non-O139 strains showed multiple antibiotic resistances in 2007 and 2012. The PCR detection of four resistance associated genes (strB, dfrA1, sulll, SXT) confirmed higher prevalence in V. cholerae non-O1/non-O139 strains. The PFGE analysis revealed 3 pulsotypes having 93% similarity among V. cholerae O1 strains., Conclusion: This study indicates the changing epidemiology, antibiogram patterns and continuous genetic variation in environmental V. cholerae strains of Odisha over the years. So continuous surveillance is necessary to understand the changing patterns of V. cholerae different serogroups isolated from stool and water samples from Odisha., (© 2023. The Author(s).)

Introduction: Cholera is a significant threat causing outbreaks/epidemics with high morbidity and mortality in coastal and tribal districts of Odisha. A sequential cholera outbreak reported from four places in Mayurbhanj district of Odisha during June to July 2009 was investigated., Methodology: Rectal swabs from diarrhea patients were analyzed for the identification, antibiogram profiles and detection of ctxB genotypes by double mismatch amplification mutation (DMAMA) polymerase chain reaction (PCR) assays and sequenced. The different virulent and drug resistant genes were detected by multiplex PCR assays. The clonality analysis on selected strains was done by pulse field gel electrophoresis (PFGE)., Results: Bacteriological analysis of rectal swabs revealed the presence of V. cholerae O1 Ogawa biotype El Tor which were resistant to co-trimoxazole, chloramphenicol, streptomycin, ampicillin, nalidixic acid, erythromycin, furazolidone and polymyxin B. DMAMA-PCR assay revealed that the cholera outbreak in Mayurbhanj district was due to both ctxB1 and ctxB7 alleles of V. cholerae O1 El Tor strains. All the V. cholerae O1 strains were positive for all virulence genes. The multiplex PCR assay on V. cholerae O1 strains revealed the presence of antibiotic resistance genes like dfrA1 (100%), intSXT (100%), sulII (62.5%) and StrB (62.5%). PFGE results on V. cholerae O1 strains exhibited two different pulsotypes with 92% similarity., Conclusions: This outbreak was a transition phase where both ctxB genotypes were prevalent after which the ctxB7 genotype gradually became dominant in Odisha. Therefore, close monitoring and continuous surveillance on diarrheal disorders is essential to prevent the future diarrheal outbreaks in this region., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2023 Bibhuti Bhusan Pal, Ashish Kumar Nayak, Smruti Ranjan Nayak, Swatishree Pany.)

The genome of Vibrio cholerae O139 strains has undergone cryptic changes since its first emergence in 1992 in South India. This study aimed to determine the presence of genotypic changes marked in ctxB, tcpA and rstR genes located within the CTX prophages among the strains of V. cholerae O139 isolated from 1999 to 2017 in Odisha. Antibiotic susceptibility test was conducted on 59 V. cholerae O139 strains. A conventional PCR assay was done for ctxB gene typing followed by sequencing along with identification of rstR and tcpA gene. Pulsed-field gel electrophoresis (PFGE) was carried out to reveal clonal variations among the V. cholerae O139 strains. Among V. cholerae O139 isolates more than 60% showed resistance to ampicillin, co-trimoxazole, furazolidone, streptomycin, neomycin and nalidixic acid. The ctxB sequencing and rstR allele-specific PCR assay revealed the presence of three genotypes 1, 3 and 4 with at least one copy of CTX Calc φ in addition to CTX ET and CTX Cl prophages in V. cholerae O139 isolates. PFGE analysis revealed 13 pulsotypes with two clades having 60% similarity among V. cholerae O139 strains. The circulating V. cholerae O139 strains in Odisha showed variation in genotypes with multiple clonal expansions over the years., (© 2021 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Staphylococcus aureus has emerged as an important pathogen among diabetic foot ulcers in patients with diabetes. Infections with S. aureus in diabetic ulcers need surveillance of resistant microbial profile to provide the basis for empirical therapy for the reduction of lower extremities amputation. Panton valentine leucocidin ( PVL ) is considered as one of the major virulence gene of S. aureus which is responsible for destruction of white blood cells and tissue necrosis. This pore forming cytotoxin gene is carried out by both methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) strains. The present study described the prevalence of PVL gene in MSSA and MRSA strains isolated from diabetic ulcer patients treated during November, 2019 to January, 2021 from a tertiary care hospital, Odisha. Infected tissue and blood samples from these patients were collected aseptically and sub-cultured using different media and standard techniques. The isolated genomic DNA of MSSA and MRSA strains were subjected to PCR assay for the detection of PVL gene. Two hundred ten S. aureus out of 402 diabetic ulcer patients were isolated having 59.52% MSSA and 40.47% MRSA strains. Wagner's grade III and grade IV ulcers were most prevalent in these ulcer patients. The prevalence of PVL gene in MSSA strains was more in comparison to MRSA strains. Forty five resistance patterns were observed from the antibiogram profiles of S. aureus. The present study highlighted that PVL gene could not be a marker for the detection of MRSA and MSSA strains in diabetic ulcer patients.

Cholera localized outbreaks/epidemics accounting for high morbidity and mortality have been reported in different years both from the coastal and tribal districts of Odisha. In the present study, the emergence and spread of two sequential cholera outbreaks reported in July to October 2012 from Rayagada and Kalahandi districts of Odisha was investigated. Environmental water samples from different sources and rectal swabs from diarrhoea patients were analysed for identification, antibiogram profiles and molecular studies using DMAMA-PCR assays. The pulsed field gel electrophoresis (PFGE) was done on some selected Vibrio cholerae O1 strains isolated from these cholera outbreak areas. Results showed 42% of rectal swabs and 2.3% of water samples collected from both the districts were positive for Vibrio cholerae O1 Ogawa biotype El Tor carrying both ctxB1 and ctxB7 genotypes. The common resistance profile of V. cholerae O1 strains was ampicillin, nalidixic acid, furazolidone and co-trimoxazole. The PFGE analysis on selected V. cholerae O1 strains of ctxB1 and ctxB7 genotypes showed three pulsotypes with 96% similarity matrix exhibiting the relationship with their respective water sources. Hence, continuous surveillance is highly essential to monitor the antibiogram profile and changing pattern of ctxB genotypes of V. cholerae O1 in this region.

The origin, spread and molecular epidemiology of altered El Tor Vibrio cholerae O1 strains isolated from cholera outbreaks/surveillance studies between 1995 and 2019 from different district of Odisha were analyzed. The stock cultures of V. cholerae O1 strains from 1995 to 2019 were analyzed through molecular analysis using different PCR assays and pulse field gel electrophoresis (PFGE) analysis. The spread map (month, year and place) was constructed to locate the dissemination of altered El Tor variants of V. cholerae O1 in this region. A total of 13 cholera outbreaks were caused by V. cholerae O1 Ogawa biotype El Tor carrying ctxB1 and ctxB7 genotypes. The ctxB1 alleles of V. cholerae O1 mostly confined to the coastal areas, whereas the ctxB7 genotypes, though originating in the coastal region of Odisha, concentrated more in the tribal areas. The positive correlation between virulence-associated genes (VAGs) was found through Pearson's correlation model, indicative of a stronger association between the VAGs. The clonal relationship through PFGE between ctxB1 and ctxB7 genotypes of V. cholerae O1 strains exhibited 80% similarity indicating single- or multi-clonal evolution. It is evident from this study that the spread of multidrug-resistant V. cholerae O1-altered El Tor was dominant over the prototype El Tor strains in this region. The origin of altered El Tor variants of V. cholerae O1 occurred in the East Coast of Odisha established that the origin of cholera happened in the Gangetic belts of Bay of Bengal where all new variants of V. cholerae O1 might have originated from the Asian countries., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer PK declared a shared affiliation with the authors to the handling editor at the time of the review, (Copyright © 2021 Pal, Behera, Nayak and Nayak.)

Purpose: Diarrheal disorders particularly cholera cause a significant threat resulting in high morbidity and mortality in the coastal and tribal areas of Odisha. Two sequential diarrheal outbreaks reported in 2016 from Balasore and Rayagada districts of Odisha were investigated to find out the causative organisms, antibiogram profile and molecular analysis of the isolated pathogens., Method: Bacteriological analysis and antibiogram profiles of the pathogens were carried out as per the standard procedure followed. The double mismatch amplification mutation (DMAMA) PCR for ctxB gene, sequencing and pulse-field gel electrophoresis (PFGE) were carried out on Vibrio cholerae O1 strains., Results: The rectal swabs and water samples from these districts were positive for V. cholerae O1 Ogawa biotype El Tor. The V. cholerae O1 strains isolated from Balasore district were multidrug resistant to many antibiotics which differed from the isolates of Rayagada district. The DMAMA PCR assay on all clinical and water isolates from these areas and some strains from other districts exhibited ctxB7 allele of V. cholerae O1 which correlates with the sequencing results having different pulsotypes. The Haitian variant of V. cholerae O1 strains which were compared with the V. cholerae O1 strains of 1999 and 2000 exhibited different pulsotypes., Conclusion: The present study reports cholera outbreaks due to multidrug resistant ctxB7 allele of V. cholerae O1 from both coastal (Balasore) and tribal (Rayagada) areas of Odisha., (Copyright © 2021 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

The present study reported the antimicrobial susceptibility trends, virulence genes, and drug resistance genes of Vibrio cholerae O1 strains isolated from outbreaks and epidemics over two and half decades (1995-2019) from Odisha, India. Antimicrobial susceptibility testing was performed by disc diffusion method. Virulence and drug resistance genes were detected by multiplex PCR assays. All V. cholerae O1 strains were sensitive to gentamicin, chloramphenicol, norfloxacin and ciprofloxacin while resistant to one or more antibiotics used. About 90% of the isolates of V. cholerae O1 carried antibiotic drug resistant genes (SulII, dfrA1 and strB) and SXT elements and the results correlated with the phenotypic antibiotic data obtained through disc diffusion assay. The tcpA Haitian variant V. cholerae O1 first appeared in 1999, gradually showing its increasing number upto 2019. TcpA El Tor strains only prevailed from 1995 to 2006; whereas the tcpA classical strains of V.choleraeO1 were found in less number from 1995 to 2016. Two multiplex PCR assays confirmed the presence of various toxigenic and virulence genes (toxR, ompU, ace, rtxC, ctxA, tcpA, rfbO1 and ompW) in all isolate of V. cholerae O1 strains. The present findings demonstrated the origin and spread of Haitian variants tcpA in V. cholerae O1 strains over two and half decades., (© 2021 Society for Applied Microbiology and John Wiley & Sons Ltd.)

The environmental reservoirs of different serogroups of Vibrio cholerae causing cholera in the flowing freshwater bodies of the tribal areas of Odisha are not known. So the present study was conducted from June 2017 to March 2020 to find out the environmental reservoirs of V. cholerae serogroups in the water and plankton samples collected from the river, nala, stream and chua from Rayagada district. Similarly, rectal swabs were collected from diarrhoea patients and correlation was established among the V. cholerae strains isolated from diarrhoea patients and environmental V. cholerae isolates through routine culture, different multiplex PCR assays and pulse field gel electrophoresis (PFGE) analysis using standard techniques. The multiplex PCR assays on biotypes and different toxic genes exhibited similar correlation between the clinical and water isolates, which was further strengthened by PFGE analysis. The planktonic DNA was positive for ctxA gene which established that the environmental water bodies were the reservoirs for virulence genes of V. cholerae serogroups. The detection of environmental reservoirs of V. cholerae serogroups in temporarily stagnant condition of water; partially encircled by stones, and near the bank of the river, nala and stream were the reservoirs which is a rare report from Odisha, India and Globe., (© 2020 Society for Applied Microbiology and John Wiley & Sons Ltd.)

This study reports variants of the ctxB allele of Vibrio cholerae O1 isolated between 1995 and 2019 in Odisha, India. ctxB1 genotypes dominated from 1995 to 2016. The Haitian variant and El Tor ctxB3 genotypes of V. cholerae O1 emerged in 1999, and were most common in 2018-2019 and 2005-2011, respectively. The ctxB7 genotype of the Haitian variant of V. cholerae O1 was quiescent from 2000 to 2006, but further spread was noted from 2007 to 2019., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Cholera posed a significant threat causing outbreaks/epidemics with high morbidity and mortality in Odisha. This study envisages the characterisation of isolated pathogen from two cholera outbreaks reported in 2018 and 2019 from Bargarh and Rayagada districts of Odisha respectively. Vibrio cholerae O1 were isolated following standard techniques. The different virulent and drug resistant genes were detected by multiplex PCR assays; whereas the ctxB genotypes were characterised through double mismatch amplification mutation (DMAMA) PCR assay. The ctxB genes were further sequenced and pulse-field gel electrophoresis (PFGE) was done on some selected strains. The clinical and water isolates of Haitian variant (HCT) V. cholerae O1 Ogawa biotype El Tor with multi drug resistant strains were isolated from both the places. All the V. cholerae O1 strains were positive for virulence genes. The antibiotic resistant genes like dfrA1 (100%), strB (76.9%), intSXT (61.5%) were detected. The PFGE results on V. cholerae O1 strains exhibited two different pulsotypes. These cholera outbreaks were due to multidrug resistant HCT variant V. cholerae O1 strains which were circulating and caused the cholera outbreaks in Odisha. So continuous surveillance on diarrheal disorders is highly essential to prevent the future diarrheal outbreaks in this region.

The Vibrio species undergo cryptic changes in their genetic material for better adaptability, which accounts for antibiotic resistance. In the present study, we investigated the emergence and spread of sensitivity to polymyxin B (PB) by El Tor V. cholerae O1 strains from 1995 to 2019 in Odisha, India. The results showed that out of 1200 V. cholerae O1 strains, 89.4% were resistant and the remaining 10.6% strains were sensitive to PB. The sensitivity to PB of V. cholerae O1 strains emerged from 2005 to 2019, except in 2015, clearly signifying the presence of classical biotype characteristics in the El Tor variant of V. cholerae O1 strains. The Etest assay revealed some interesting traits of PB susceptibility in the ctxB1 and ctxB7 genotypes of V. cholerae O1 strains. The minimum inhibitory concentration (MIC) of ctxB7 genotypes showed reduced MIC values of ≤ 4 µg/mL, whereas ctxB1 genotypes exhibited higher MIC values of 24 and 32 µg/mL.

Infectious diarrheal diseases remain a leading cause of morbidity and mortality in developing and underdeveloped countries. The present study documented the etiology of bacterial enteropathogens in three tribal districts of Odisha from July 2010 to September 2013. A total of 1427 rectal swabs were collected and bacteriologically analyzed by following standard procedure. Among the 930 (65.2%) culture positive samples, Escherichia coli (E. coli) constituted 636 (44.6%); Vibrio cholerae (V. cholerae) O1, 146 (10.2%); Salmonella species (spp.), 10 (0.7%); Shigella spp., 79 (5.5%); and Aeromonas spp., 59 (4.1%). Of the 729 environmental water samples taken from river, open well, Nala (a small stream), and Chua (a shallow pit on a river bed), 14 (1.9%) contained non-O1/non-O139 V. cholerae and 13 (1.8%) had V. cholerae O1 strains. An analysis of the demographics showed that people in the 14 to 40-year age group were highly susceptible to diarrhea caused by V. cholerae which occurred mainly during the rainy and post-rainy seasons. All enteropathogens were multidrug-resistant and found throughout the study period. The V. cholerae strains isolated were El Tor variants carrying the classical, El Tor, and Haitian cholera toxin subunit B (ctxB) genes. The classical ctxB was the dominant allele, and the prevalence of the Haitian ctxB allele increased during the test period. These findings indicate that active surveillance is needed to monitor the changing antibiotic resistance patterns of V. cholerae serogroups and biotypes present in this region.

Gram-negative bacteria is the main causative agents for columnaris disease outbreak to finfishes. The outer membrane proteins (OMPs) candidate of Flavobacterium columnare bacterial cell served a critical component for cellular invasion targeted to the eukaryotic cell and survival inside the macrophages. Therefore, OMPs considered as the supreme element for the development of promising vaccine against F. columnare . Implies advanced in silico approaches, the predicted 3-D model of targeted OMPs were characterized by the Swiss model server and validated through Procheck programs and Protein Structure Analysis (ProSA) web server. The protein sequences having B-cell binding sites were preferred from sequence alignment; afterwards the B cell epitopes prediction was prepared using the BCPred and amino acid pairs (AAP) prediction algorithms modules of BCPreds. Consequently, the selected antigenic amino acids sequences (B-cell epitopic regions) were analyzed for T-cell epitopes determination (MHC I and MHC II alleles binding sequence) performing the ProPred 1 and ProPred server respectively. The epitopes (9 mer: IKKYEPAPV, YGPNYKWKF and YRGLNVGTS) within the OMPs binds to both of the MHC classes (MHC I and MHC II) and covered highest number of MHC alleles are characterized. OMPs of F. columnare being conserved across serotypes and highly immunogenic for their exposed epitopes on the cell surface as a potent candidate focus to vaccine development for combating the disease problems in commercial aquaculture. The portrayed epitopes might be beneficial for practical designing of abundant peptide-based vaccine development against the columnaris through boosting up the advantageous immune responses.Communicated by Ramaswamy H. Sarma.

Cholera is an acute diarrheal disease caused by Vibrio cholerae serogroups O1 and O139, which are known to cause epidemics of cholera in Odisha. The present study was intended to document the antibiotic resistance pattern among clinical isolates of both serogroups of V. cholerae (O1 and O139) isolated during 2004-2013. Nine-hundred nine isolates of V. cholerae were included in this study and were identified by standard procedures. An antibiotic sensitivity test was performed by the disc diffusion method. The seasonality of cholera in this region indicated that there was one peak in the rainy season only. The number of cholera cases started increasing from July and declined starting from the month of October onward. The adult age group of patients was the worst affected among all age groups of patients. The 2 different serogroups of V. cholerae (O1 and O139) showed different prevalence rates (%) of resistance to all the antibiotics in each year. Serogroup O1 showed uniformly high resistance to co-trimoxazole, furazolidone, and nalidixic acid throughout the study. Chloramphenicol encountered resistance only during 2009, but the strains were sensitive in the other years. The emergence of multiple drug-resistant V. cholerae strains may significantly influence the control of future outbreaks and epidemics of cholera in this region.

The large outbreak of cholera reported during July to September 2014 in the Narla block of Kalahandi district, India, was investigated to determine the causative organism. Rectal swabs collected from patients with diarrhea and environmental water samples were cultured following standard techniques. The causative organism was identified as Vibrio cholerae O1 Ogawa biotype El Tor, and analysis by double mismatch mutation assay PCR confirmed that all strains were the ctxB7 variant of Haitian V. cholerae O1. The environmental water samples were negative for V. cholerae. The V. cholerae O1 strains were sensitive to tetracycline, ciprofloxacin, norfloxacin, ofloxacin, doxycycline, and azithromycin, but were resistant to erythromycin, gentamicin, chloramphenicol, furazolidone, neomycin, cotrimoxazole, nalidixic acid, and ampicillin. In the 2014 cholera outbreak, the early reporting of the pathogen enabled the government authorities to implement adequate control measures in time to curtail the spread of the disease. That was the second large cholera outbreak due to Haitian variants of V. cholerae O1 after the 2010 Haiti cholera outbreak reported from Odisha, India, and other locations globally. Active surveillance is required to track the spread of this strain in the Odisha region.

Objectives: The epidemics of cholera were reported in the Kashipur, K.singhpur, B cuttack blocks of Rayagada district and Mohana block of Gajapati district of Odisha during 2010. The present study was carried out to isolate the bacterial pathogen, its drug sensitivity pattern and to describe the spread of the disease in those areas., Methods: A total of 68 rectal swabs collected from patients with severe diarrhea, admitted to different health centers and diarrhea affected villages were bacteriologically analyzed. Similarly 22 water samples collected from different villages from nala, chua, etc were tested for the presence of V cholerae., Results: Out of 68 rectal swabs tested 35 (51.5%) were V cholerae O1 Ogawa and 14(20.6%) were E coli; which might be commensals. All water samples were negative for V cholerae. The V cholerae strains were sensitive to gentamicin, norfloxacin, ciprofloxacin, azithromycin and ofloxacin; but were resistant to ampicillin, tetracycline, nalidixic acid, furazolidone, streptomycin, erythromycin, co-trimoxazole, neomycin and chloramphenicol. All V cholerae strains were 100% resistant to tetracycline and they were El Tor variants harboring ctxB gene of classical strain., Conclusions: The present study indicated the emergence and spread of tetracycline resistant V cholerae O1 El Tor variant in the tribal areas which needs close monitoring., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Glial cells are very important for normal brain function and alterations in their activity due to hyperglycemia, could contribute to diabetes-related cognitive dysfunction. Oxidative insults often cause rapid changes in almost all cells including glial cells. However, pathophysiologic mechanisms that lead to diabetic complications are not completely elucidated. Therefore, we examined whether elevated glucose levels directly or indirectly disrupt antioxidant defense mechanisms causing alterations in signaling pathways, cell cycle dysregulation, and reactive oxygen/nitrogen species-mediated apoptosis in glial cells. Findings of this study demonstrated that exposure of glial cells to high glucose markedly induces cellular and molecular injuries, as evidenced by elevated levels of reactive oxygen/nitrogen species, biomolecules damage, cell cycle dysregulation, decrease in antioxidant enzymes, and decrease in cell viability. Pretreatment of cells with N-acetyl-L-cysteine reduced high glucose-induced cytotoxicity by increasing the levels of antioxidant enzymes, and decreasing the number of apoptotic cells. Further, at molecular level high glucose treatment resulted in a significant increase in phosphorylation of Akt, MAPKs, tuberin, down regulation of 8-oxoG-DNA glycosylase and increase in 8-hydroxydeoxyguanosine accumulations. Pretreatment of cells with N-acetyl-L-cysteine, phosphatidylinositol3-kinase/Akt and ERK1/2 inhibitors completely abolished the apoptotic effects of high glucose. Moreover, N-acetyl-L-cysteine significantly inhibited reactive oxygen/nitrogen species generation, elevated antioxidants levels, inhibited Akt, ERK1/2, tuberin phosphorylation, decreased 8-hydroxydeoxyguanosine accumulation and upregulated 8-oxoG-DNA glycosylase expression. Our results demonstrate that high glucose induces apoptosis and inhibits proliferation of glial cells, which may be mediated by the phosphorylation of tuberin, down regulation of 8-oxoG-DNA glycosylase and 8-hydroxydeoxyguanosine accumulation via activation of Akt and ERK1/2MAPK pathways., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A large outbreak of cholera reported during April-July 2009 in the Kendrapada district of Odisha, India was investigated. Forty-one rectal swabs and 41 water samples, collected from diarrhoeal patients and from different villages were bacteriologically analysed for the isolation of bacterial enteriopathogens, antibiogram profile and detection of various toxic genes. The bacteriological analysis of rectal swabs and environmental water samples revealed the presence of V. cholerae O1 Ogawa biotype El Tor. The V. cholerae strains were resistant to ciprofloxacin, co-trimoxazole, chloramphenicol, streptomycin, ampicillin, furazolidone and nalidixic acid. The multiplex polymerase chain reaction (PCR) assay on V. cholerae strains revealed the presence of ctxA and tcpA genes. The mismatch amplification of mutation assay (MAMA) PCR on clinical and environmental isolates of V. cholerae revealed that the strains were El Tor biotype, which harboured the ctxB gene of the classical strain. The random amplified polymorphic DNA PCR analysis and pulsed-field gel electrophoresis results indicated that the V. cholerae isolates belonged to the same clone. This investigation gives a warning that the El Tor variant of V. cholerae has spread to the coastal district causing a large outbreak that requires close monitoring and surveillance on diarrhoeal outbreaks in Odisha.

The emergence and spread of Vibrio cholerae O1 El Tor variant strains causing severe diarrhea has been witnessed worldwide in recent years. In the state of Odisha, India, the spread of the V. cholerae O1 El Tor variant strains was studied during outbreaks in 2008 and 2009. Analysis of 194 V. cholerae O1 Ogawa strains revealed that V. cholerae O1 El Tor variant strains are spreading gradually throughout the state, causing outbreaks replacing typical V. cholerae O1 El Tor biotype strains.

Background: We investigated the epidemic of cholera that occurred in Kashipur and Dasmantpur blocks of Orissa, reported during July-September 2007., Methods: Sixty-two rectal swabs and 28 water samples collected from diarrhea patients at different hospitals and villages were bacteriologically analyzed for the identification, antibiogram, and detection of toxic genes of Vibrio cholerae., Results: The cholera outbreaks were caused by V. cholerae O1 Ogawa biotype El Tor in both Kashipur and Dasmantpur blocks. All the V. cholerae isolates from the clinical and environmental samples were sensitive to tetracycline, gentamicin, azithromycin, and chloramphenicol, but were resistant to ampicillin, ciprofloxacin, norfloxacin, co-trimoxazole, nalidixic acid, neomycin, and furazolidone, except the water isolates, which were sensitive to ciprofloxacin and norfloxacin. The multiplex PCR assay revealed that all the clinical and environmental V. cholerae isolates were positive for the ctxA and tcpA genes, showing biotype El Tor. Interestingly, 88% of the clinical and environmental isolates of V. cholerae were El Tor biotype with mutation at the ctxB gene of the classical strain, as confirmed by mismatch amplification of mutation (MAMA)-PCR assay., Conclusions: This is the first report of the El Tor variant of V. cholerae O1 Ogawa having the ctxB gene of the classical strain with altered antibiogram causing epidemics of cholera in Orissa, India., (Copyright 2009 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background/purpose: Cholera has been reported in the state of Orissa, India during the last decades. An explosive outbreak of diarrhea occurred in Central Cuttack Ward 22 of Orissa (population approximately 10,621), between March 12-23, 2006. This outbreak was investigated by a team from the Regional Medical Research Centre of Bhubaneswar to identify the causative agents and to determine the antimicrobial susceptibility pattern and associated virulent genes., Methods: Clinical and epidemiological data were collected from 100 hospitalized patients with diarrhea from the Sriram Chandra Bhanja Medical College, Cuttack, Orissa. Rectal swabs and water samples were collected and tested for diarrheagenic enteropathogens. Isolated Vibrio cholerae were subjected to antibiotic susceptibility tests and polymerase chain reaction analysis for the detection of virulent genes., Results: Of the 23 rectal swabs collected, 19 (82.6%) were positive for V. cholerae serogroup O1, serotype Ogawa. All strains were uniformly susceptible to ampicillin, gentamicin, chloramphenicol, ciprofloxacin, norfloxacin, neomycin, and tetracycline, but resistant to co-trimoxazole, furazolidone, nalidixic acid, and streptomycin. Polymerase chain reaction revealed that all strains had ctxA, tcpA (biotype El Tor), zot, and ace genes, suggesting their possible role in the outbreak., Conclusion: This is the first localized outbreak of V. cholerae O1, serotype Ogawa, in the state of Orissa in 2006 after a gap of 6 months dominated by Inaba strains., (Copyright 2010 Taiwan Society of Microbiology. Published by Elsevier B.V. All rights reserved.)

We investigated two sequential outbreaks of severe diarrhoea in two neighbouring villages of Orissa, in 2005. We conducted descriptive and matched case-control studies. The attack rates were 5.6% (n=62) and 5.2% (n=51), respectively, in the first and second villages. One death was reported in the second village (case fatality 2%). We identified that consumption of milk products prepared in the household of the index case [matched odds ratio (mOR) 5.7, 95% confidence interval (CI) 1.7-30] in the first village, and drinking well water in the second village were associated with the illness (mOR 4.7, 95% CI 1.6-19). We isolated Vibrio cholerae El Tor O1 Ogawa from stool samples from both the villages. Mishandling of milk products led to a cholera outbreak in the first village, which led to sewerage contamination of a well and another outbreak in the second village. Environmental contamination should be expected and prevented during cholera outbreaks.

Bacteriological analysis of 1,551 stool/rectal swabs from all age groups of diarrhea patients of different hospitals of Orissa from January 2004 to December 2006 was carried out using standard procedures. Among all enteropathogens isolated in 886 culture-positive samples, Escherichia coli constituted 75.5%, including 13.2% pathogenic E. coli; Vibrio cholerae O1 constituted 17.3%; V. cholerae O139, 1%; Shigella spp., 4.5% (Shigella flexneri type 6, 2.9%, S. dysenteriae type I, 0.7%, S. sonnei, 0.6%, and S. boydii, 0.3%); Salmonella spp., 0.7%; and Aeromonas spp., only 2.0%. The isolation of bacterial enteropathogens was highest during July, 2005, followed by September, 2006. The prevalence of shigellosis in this region was relatively low. Cholera cases were more frequent during the rainy seasons. The dominance of V. cholerae O1 Inaba over Ogawa serotypes was observed in 2005, whereas this trend was reversed in 2006. The resistance profile of V. cholerae O1 was co-trimoxazole (Co), furazolidone (Fr), and nalidixic acid (Na); for Aeromonas spp., it was ampicillin (A), Fr, ciprofloxacin (Cf), Na, norfloxacin (Nx), and Co. Pathogenic E. coli strains were resistant to A, Fr, Co, streptomycin (S), Cf, Na, Nx, and neomycin (N); Shigella spp. were resistant to Fr, Na, Co, and S; and Salmonella spp. were resistant to A and Fr. Active surveillance should be continued among diarrhea patients to look for different enteropathogens and to define the shifting antibiogram patterns in this region.

A quadruplex PCR was developed for the simultaneous detection of genes specific for Vibrio cholerae O1 and/or O139 serogroup (wbe and/or wbf), cholera toxin A subunit (ctxA), toxin-coregulated pilus (tcpA), and central regulating protein ToxR (toxR) in a single tube reaction. This is a simple, rapid, and accurate approach for the detection of toxigenic V. cholerae O1 and/or O139 and can prevent the rapid spread of the disease by early detection.

A total of 431 rectal swabs, collected from acute diarrheal cases at a surveillance site and at different diarrheal outbreak areas of Orissa from May to October 2005, were bacteriologically analyzed. Out of 265 culture-positive samples, Vibrio cholerae O1 was isolated in 56 samples (20.8%), of which 37 were the Inaba serotype and 19 were the Ogawa. The antibiogram profile revealed that all the V. cholerae O1 Ogawa and Inaba serotypes were uniformly sensitive to ampicillin, chloramphenicol, gentamicin, ciprofloxacin, norfloxacin and tetracycline. The V. cholerae O1 Inaba serotypes were resistant to furazolidone and nalidixic acid, while the Ogawa strains were resistant to furazolidone, nalidixic acid and neomycin. The multiplex polymerase chain reaction (PCR) assay on some selected strains of both serotypes revealed that all the strains were positive for ctxA and tcpA genes showing biotype El Tor. The present study revealed the emergence of V. cholerae O1 biotype El Tor serotype Inaba, which caused sporadic outbreaks of cholera in 2005. The outbreaks of diarrheal disorders in one geographical area of the state (in the Pattamundai area, Kendrapara district) in 2005 were due to V. cholerae O1 Ogawa, whereas the other outbreaks in other areas (Puri, Khurda and Dhenkanal districts) from August to October 2005 were due to V. cholerae O1 serotype Inaba. This is the first report that an emergence of V. cholerae O1 serotype Inaba caused sporadic outbreaks of cholera in different parts of Orissa. Switching over of V. cholerae O1 Ogawa strains to Inaba, causing diarrheal outbreaks in Orissa, needs close monitoring.

An epidemiological study was carried out to find out the aetiological agent for diarrhoeal disorders in the cyclone and flood affected areas of Orissa, India. Rectal swabs collected from 107 hospitalized diarrhoea patients were bacteriologically analysed to isolate and identify the various enteropathogens. Detection of toxic genes among E. coli and V. cholerae was carried out by polymerase chain reaction (PCR) assay. Of the 107 rectal swabs analysed, 72.3% were positive for V. cholerae O1 Ogawa, 7.2% for V. cholerae O139, 1.2% for E. coli (EAggEC) and 1.2% for Shigella flexneri type 6. Using multiplex PCR assay it was found that all V. cholerae isolates were ctxA positive and El Tor biotype. Strains of V. cholerae O1 were observed to be resistant to nalidixic acid, furazolidone, streptomycin, co-trimoxazole and ampicillin. Except for nalidixic acid, the resistance pattern for O139 was identical to that of O1 strains. Representative strains of V. cholerae were further characterized by randomly amplified polymorphic DNA (RAPD) analysis and ribotyping. Both O1 and O139 V. cholerae strains exhibited the R3 pattern of ribotype and belonged to a similar pattern of RAPD compared with that of Calcutta strains. Early bacteriological and epidemiological investigations have revealed the dominance of V. cholerae O1 among the hospitalized patients in cyclone affected areas of Orissa. Drinking water scarcity and poor sanitation were thought to be responsible for these diarrhoeal outbreaks. Timely reporting and implementation of appropriate control measures could contain a vital epidemic in this area.

Background: Helicobacter pylori (H. pylori) has been etiologically linked with primary gastric lymphoma (PGL) and gastric carcinoma (GC). There are a few reports of occurrence of both diseases in the same patients with H. pylori infection., Case Presentation: We report a patient with PGL in whom the tumor regressed after surgical resection combined with eradication of H. pylori infection. However, he developed GC on follow up; this was temporally associated with recrudescence/re-infection of H. pylori. This is perhaps first report of such occurrence., Conclusions: Possible cause and effect relationship between H. pylori infection and both PGL and GC is discussed. This case also documents a unique problem in management of PGL in tropical countries where re-infection with H. pylori is supposed to be high.

Objectives and Method: Forty patients (mean age 45 years; 24 men) attending a tertiary care hospital in eastern India during the period 1996-2000 were investigated to evaluate the etiology and clinical spectrum of obscure gastrointestinal bleed., Results: The patients presented to hospital after mean symptom duration of 2.5 years. They had received an average of 15 units of blood transfusion. Most patients presented with recurrent melena (85%); all had iron-deficiency anemia. A total of 230 investigations (89 gastroscopies, 54 colonoscopies, 25 double-contrast meal and follow-through studies, 14 small bowel enemas, 24 radionuclide scans, 16 mesenteric angiographies and 8 intraoperative endoscopies) yielded positive diagnosis in 87.5% of cases. The diseases encountered were small bowel and colonic angiodysplasias (32.5%), ileal Crohn's disease (20%), intestinal tuberculosis (10%), intestinal tumors (10%), nonspecific small bowel ulcers and strictures (7.5%), Meckel's diverticulum (5%) and hemobilia (2.5%). The etiology remained obscure in 5 (12.5%) cases. Overall success of surgery was 63%; in-hospital mortality was 7.5%., Conclusion: Though obscure gastrointestinal bleed is commonly caused by angiodysplasias, it can be an atypical presentation of Crohn's disease.

Till date only three series of immunoproliferative small intestinal disease (IPSID) describing 22 patients have been reported from India. Seven patients with IPSID in two tertiary referral centers in India are included in the study. Diagnosis was based on typical clinical features [diarrhoea (7/7), weight loss (7/7), clubbing (6/7), fever (3/7), abdominal pain and lump (3/7)], biochemical evidence of malabsorption and duodenal biopsy findings. All patients were young males (mean age 29.8 +/- 11.8 years, range 17-53). Atypical features included gastric involvement (1/7), colonic involvement (1/7) and appearance of pigmented nails following anti-cancer chemotherapy (1/7) which disappeared six months after omitting doxorubin from chemotherapy regimen. Parasitic infestation was common. Ascaris lumbricoides (1/7), Giardia lamblia and hookworm (1/7), Strongyloides stercoralis and Trichuris trichura (1/7). In the latter patient S. stercoralis became disseminated after anti-malignant chemotherapy. One patient had gastric H. pylori infection. Four of the seven patients who were misdiagnosed as tropical sprue were treated with tetracycline. This raises doubt on efficacy of tetracycline alone in treatment of IPSID. One other patient was misdiagnosed and treated as intestinal tuberculosis. Early diagnosis and administration of chemotherapy may improve survival in this disease.

Forty one consecutive patients with portal hypertension (PHT) were evaluated by colonoscopy to study the prevalence, type, extent and predictors of haemorrhoids, colorectal varices, and portal hypertensive colopathy. Specific inquiry and regular follow-up assessed frequency of hematochezia. Twenty five patients with obscure gastrointestinal bleeding without PHT who underwent colonoscopy served as controls. Haemorrhoids were seen in nine of 41 (21.9%) patients with PHT and four of 25 (16%) controls (p = ns). Colorectal varices were seen in 13/41 (31.7%) patients with PHT and none of the controls (p = 0.005). Portal colopathy was present in 15/41 (36.6%) patients with PHT and none of the controls (p = 0.0005). None of the parameters (e.g. aetiology of PHT, Child's class, oesophageal variceal eradication by EST with or without EVL, history of variceal bleeding, grade of oesophageal varices, presence of portal hypertensive gastropathy or gastric varices) predicted the occurrence of colorectal varices and portal hypertensive colopathy. Detection of colorectal varices but not portal hypertensive colopathy was associated with occurrence of hematochezia.

As the few randomized controlled trials available in the literature comparing botulinum toxin (BT) injection with established endoscopic treatment of achalasia cardia, i.e. pneumatic dilatation, showed conflicting results, we conducted a prospective randomized trial. Seventeen consecutive patients with achalasia cardia diagnosed during a period between December 1997 and February 2000 were randomized into two treatment groups [pneumatic dilatation by Rigiflex dilator (n=10), BT injection by sclerotherapy needle into four quadrants of lower esophageal sphincter (LES) (n=7) 80 units in five cases, 60 units in two cases] after dysphagia grading, endoscopy, barium esophagogram, and manometry, all of which were repeated 1 week after treatment. Patients were followed up clinically for 35.2+/-14 weeks. Chi-squares, Wilcoxon rank-sum test, Kaplan-Meier method and log-rank tests were used for statistical analysis. After 1 week, 6/7 (86%) BT-treated vs. 8/10 (80%) dilatation-treated patients improved (P=NS). There was no difference in LES pressure and maximum esophageal diameter in the barium esophagogram in the two groups before therapy. Both therapies resulted in significant reduction in LES pressure. The cumulative dysphagia-free state using the Kaplan-Meier method decreased progressively in BT-treated compared with dilatation-treated patients (P=0.027). Two patients with tortuous megaesophagus, one of whom had failed dilatation complicated by perforation previously, improved after BT. One other patient in whom pneumatic dilatation had previously failed improved in a similar manner. BT is as good as pneumatic dilatation in achieving an initial improvement in dysphagia of achalasia cardia. It is also effective in patients with tortuous megaesophagus and previous failed pneumatic dilatation. However, dysphagia often recurs during 1-year follow up.

Aim: To evaluate the aetiologic spectrum of chronic liver disease (CLD) in a tertiary referral center in Eastern India., Methods: A total of 175 patients (cirrhosis 166, chronic hepatitis 9) diagnosed by clinical, biochemical, radiological and histopathological (42 cases) parameters were evaluated for aetiology. Investigations included: HBsAg and anti HCV (third generation) by ELISA. HBeAg and HBV DNA were tested in HBsAg positive patients. HCV RNA was tested in anti-HCV positive patients. Markers for autoimmune and Wilson's disease (anti-nuclear antibody, anti smooth muscle antibody, serum ceruloplasmin, urinary copper and slit lamp examination for KF ring) were done where clinically indicated., Results: A total of 62 (35.4%) patients had HBV related CLD and 6 (9.7%) of them had pre-core mutant. HCV was present in 17/114 (14.9%) cases and none had infection with both viruses. Autoimmunity, Wilson's disease and alcohol were the aetiological factors in 5 (2.8%), 5 (2.8%) and 3 (1.7%) patients respectively. No aetilogy could be found in 18/114 (15.8%) patients., Conclusions: It is concluded that HBV is the commonest cause of CLD in Eastern India. Alcohol and HCV are uncommon in this part of the country.

Endoscopic sclerotherapy (EST) leads to structural and motility changes in the esophagus; the former are thought to be commoner after EST with absolute alcohol (AA), which is a commonly used sclerosant in India as it is cheap and effective. There are no previous studies on changes in esophageal motility after EST with AA. Accordingly, we studied patients with portal hypertension before (n = 24) and after (n = 22) variceal obliteration by EST with AA using a water perfusion esophageal manometry system. Contraction amplitude in the distal esophagus was reduced in the post-EST group compared with the pre-EST group (63.4 +/- 24.9 vs. 18.2 +/- 14.3 mmHg, p < 0.01). Duration of esophageal contraction in both the proximal and distal esophagus became prolonged in the post-EST compared with the pre-EST group (3.3 +/- 0.8 vs. 5.4 +/- 2.6 and 4.3 +/- 1.1 vs. 6.6 +/- 2.3 s, p < 0.001 for both). Lower esophageal sphincter (LES) pressure was reduced in the post-EST compared with the pre-EST group, although the difference was not significant statistically. Abnormal contraction waveforms were more frequent in the post-EST group. One patient in the post-EST group had persistent dysphagia in the absence of endoscopically documented stricture at the time of manometric study. This study shows frequent occurrence of esophageal dysmotility after EST with AA; however, esophageal dysmotility after EST was infrequently associated with motor dysphagia.

Seroprevalence of human immunodeficiency virus (HIV) infection was carried out in three prisons in Orissa from March 1994 to December 1995. All the prisoners of Indian origin (300), housed in these jails tested negative for HIV infection. On the other hand, 33.8 per cent (26/77) of jail inmates from foreign countries (Thailand and Myanmar), serving short terms in Orissa jails were found positive for HIV infection. While all the 26 HIV positive prisoners had HIV-1 infection, five of them had antibodies for HIV-1 and HIV-2 viruses. Such a high prevalence of HIV infection and detection of HIV-2 virus is being reported for the first time from Orissa.

The genotoxic potential of zinctox (zinc phosphide) was evaluated in mice in vivo employing different bioassays: bone marrow chromosome aberration, micronucleus and sperm shape abnormality. A significant increase of chromosome aberrations was induced by three doses. Interestingly, the highest percentage was induced by the middle dose. In a route-responsive study the highest percentage of chromosome aberration was induced by the oral route. A dose-dependent increase in the incidence of micronuclei was noticed in the bone marrow cells. In the sperm shape abnormality test a high percentage of abnormal sperm was induced by all three dose levels. The results revealed the genotoxic potential of zinctox in the tested mouse in vivo system.

Cryptosporidium infection was found in association with acute diarrhoeal disease in ten patients (13%) of a total of 77 children below the age of 8 years admitted over a period of 1 year to Bhubaneswar Capital Hospital, Orissa state, India. Contrary to another report from South India, there were no asymptomatic cryptosporidium oocyst excretors detected either in the matched control group of 42 children without diarrhoea or in a group of 113 healthy school children investigated. A clustering of seven cases occurred in the months of the rainy season and of three cases in the winter months. Nine out of 63 children (14.3%) below the age of 2 years were infected. Seven of the ten infected children were significantly below their expected weight-for-age, but the mean weight-for-age did not differ from those of the uninfected children. The possible epidemiological factors in the transmission of cryptosporidium are discussed.