Neglected malaria parasites in hard-to-reach areas of Odisha, India: implications in elimination programme.

BACKGROUND: Information on the foci of Plasmodium species infections is essential for any country heading towards elimination. Odisha, one of the malaria-endemic states of India is targeting elimination of malaria by 2030. To support decision-making regarding targeted intervention, the distribution of Plasmodium species infections was investigated in hard-to-reach areas where a special malaria elimination drive, namely Durgama Anchalare Malaria Nirakaran (DAMaN) began in 2017. METHODS: A cross-sectional survey was conducted in 2228 households during July to November 2019 in six districts, to evaluate the occurrence of Plasmodium species. The species were identified by polymerase chain reaction (PCR) followed by sequencing, in case of Plasmodium ovale. RESULTS: Of the 3557 blood specimens tested, malaria infection was detected in 282 (7.8%) specimens by PCR. Of the total positive samples, 14.1% were P. ovale spp. and 10.3% were Plasmodium malariae infections. The majority of P. ovale spp. (75.8%) infections were mixed with either Plasmodium falciparum and/or Plasmodium vivax and found to be distributed in three geophysical regions (Northern-plateau, Central Tableland and Eastern Ghat) of the State, while P. malariae has been found in Northern-plateau and Eastern Ghat regions. Speciation revealed occurrence of both Plasmodium ovale curtisi (classic type) and Plasmodium ovale wallikeri (variant type). CONCLUSIONS: In the present study a considerable number of P. ovale spp. and P. malariae were detected in a wide geographical areas of Odisha State, which contributes around 40% of the country's total malaria burden. For successful elimination of malaria within the framework of national programme, P. ovale spp. along with P. malariae needs to be incorporated in surveillance system, especially when P. falciparum and P. vivax spp. are in rapid decline.

Decades of cholera in Odisha, India (1993-2015): lessons learned and the ways forward.

Cholera is one of the major public health problems in the state of Odisha, India since centuries. The current paper is a comprehensive report on epidemiology of cholera in Odisha, which was documented from 1993. PubMed and Web of Knowledge were searched for publications reporting cholera in Odisha during the period 1993-2015. The search was performed using the keywords 'Odisha' and/or 'Orissa' and 'Cholera'. In addition, manual search was undertaken to find out relevant papers. During the study period, a total of 37 cholera outbreaks were reported with an average of >1.5 cholera outbreaks per year and case fatality ratio was 0.3%. Vibrio cholerae O1 Ogawa serotype was the major causative agent in most of the cholera cases. The recent studies demonstrated the prevalence of V. cholerae O1, El Tor variants carrying ctxB1, ctxB7 and Haitian variant tcpA allele associated with polymyxin B sensitivity and these variants are replacing the proto type El Tor. The first report of variant ctxB7 in Odisha during super-cyclone 1999 predicted its emergence and subsequent spread causing cholera outbreaks. The prevalence of multidrug-resistant V. cholerae at different time periods created alarming situation. The efficacy trial of oral cholera vaccine (OCV, Shanchol) in a public health set-up in Odisha has shown encouraging results which should be deployed for community level vaccination among the vulnerable population. This paper has taken an effort to disseminate the valuable information of epidemiology of cholera that will influence the policy-makers and epidemiologists for constant surveillance in other parts of Odisha, India and around the globe.

Identification of population of bacteria from culture negative surgical site infection patients using molecular tool.

BACKGROUND: Managing surgical site infections, with negative culture report in routine diagnosis is a common dilemma in microbiology accounting more than 30% worldwide. The present study attempted to identify the presence of bacterial spp. if any in wound aspirates/swabs of culture negative surgical site infections of hospitalised patients using molecular tools. METHODS: Ninety-seven patients with post-operative SSI whose wound swabs/aspirate were negative in the conventional aerobic culture after 72 h of incubation were analysed by 16S rRNA gene specific broad range PCR. The amplified DNA fragments were sequenced by Sanger DNA sequencing method and homology of the sequence were matched using NCBI BLAST (NCBI, USA) RESULTS: Of the 97 patients, 16S rRNA based broad range PCR assay could identify the presence of bacterial pathogen in 53(54.63%) cases, of which 29 isolates were supposed to be of viable but non-culturable bacteria (VBNC), 07 were of obligatory anaerobes and 13 were of unculturable bacteria, 04 were with poly bacterial infections. CONCLUSIONS: Our study highlights the usefulness of PCR assay in detecting the presence of any VBNC, anaerobes and unculturable bacteria in SSI patients regardless of how well the bacteria may or may not grow in culture. Measures should be taken to use anaerobic culture system and PCR diagnosis along with conventional culture to detect the VBNC and unculturable bacteria where Gram stain is positive for better patient care.

Malnutrition and Anemia Among Particularly Vulnerable Tribal Groups of Odisha, India: Needs for Context-Specific Intervention.

Background: As undernutrition and anemia persist to be prevalent in India, the socioeconomically disadvantaged groups continue to take the greater brunt. Odisha is home to the largest number of particularly vulnerable tribal groups (PVTGs) in India. The study aimed to provide a comprehensive report on the undernutrition and anemia status of all the PVTGs of Odisha. Methods: A community-based cross-sectional study was conducted among (N = 1461, 683 males and 779 females) 13 PVTGs spread across 12 districts of Odisha from August 2018 to February 2019. Results: Among the under-five children, the prevalence of underweight was observed in 75.26%, stunting in 55.42%, and wasting in 60.00% and all forms of undernutrition were higher among girls. Among children and adolescents belonging to the age group of 5 to 19 years, the prevalence of thinness was 46.7%. In individuals above the age of 20, the prevalence of underweight among males was 37.7% and females was 44.3% and severe anemia was present in 36.5% of females and 35.8% of males. Women in the reproductive age have a higher prevalence of anemia. Conclusion: The study shows that undernutrition and anemia remain high in the PVTGs, especially among the under-five children and women in the reproductive age. As the country heads toward fulfilling Sustainable Development Goals (SDG) by 2030, national and state health policies need to be designed and implemented, giving special focus to these vulnerable groups.

Assessment of effectiveness of DAMaN: A malaria intervention program initiated by Government of Odisha, India.

India, a persistently significant contributor to the global malaria burden, rolled out several anti-malaria interventions at the national and state level to control and recently, to eliminate the disease. Odisha, the eastern Indian state with the highest malaria burden experienced substantial gains shown by various anti-malaria initiatives implemented under the National Vector-borne Disease Control Programme (NVBDCP). However, recalcitrant high-transmission "pockets" of malaria persist in hard-to-reach stretches of the state, characterised by limited access to routine malaria surveillance and the forested hilly topography favouring unbridled vector breeding. The prevalence of asymptomatic malaria in such pockets serves as perpetual malaria reservoir, thus hindering its elimination. Therefore, a project with the acronym DAMaN was initiated since 2017 by state NVBDCP, targeting locally identified high endemic 'pockets' in 23 districts. DAMaN comprised biennial mass screening and treatment, provisioning of long-lasting insecticidal net (LLIN) and behavioural change communication. Subsequently, to inform policy, assessment of DAMaN was conceived that aims to estimate the coverage of the various components of the project; the prevalence of malaria, even at sub-patent level especially among pregnant/lactating women and children; and its impact on malaria incidence. A survey of DAMaN beneficiaries will measure coverage; and knowledge and practices related to LLIN; along with collection of blood specimens from a probability sample. A multi-stage stratified clustered sample of 2228 households (~33% having pregnant/lactating women) will be selected from 6 DAMaN districts. Routine DAMaN project data (2017-2018) and NVBDCP data (2013-2018) will be extracted. Rapid Diagnostic Test, Polymerase Chain Reaction and blood smear microscopy will be conducted to detect malarial parasitemia. In addition to measuring DAMaN's coverage and malarial prevalence in DAMaN pockets, its impact will be estimated using pre-post differences and Interrupted Time Series analysis using 2017 as the "inflection" point. The assessment may help to validate the unique strategies employed by DAMaN.

Sequence Analysis of the K13-Propeller Gene in Artemisinin Challenging Plasmodium falciparum Isolates from Malaria Endemic Areas of Odisha, India: A Molecular Surveillance Study.

Estimation of the spread and advancement of Plasmodium falciparum artemisinin-resistant parasites can be done by probing polymorphisms in the kelch (Pfk13) domain (a validated molecular marker). This study aimed to provide baseline information for future artemisinin surveillance by analyzing the k13-propeller domain in P. falciparum field isolates collected from 24 study areas in 14 malaria hot spots of Odisha (previously Orissa) during July 2018-January 2019. A total of 178 P. falciparum mono infections were assessed. An 849-base pair fragment encoding the Pfk13 propeller was amplified by nested polymerase chain reaction and sequenced in both directions (PCR). After DNA alignment with the 3D7 reference sequence, all samples were found to be wild type. It can be anticipated that malaria public health is not under direct threat in Odisha relating to ART resistance.

A prospective study on incidence, risk factors, and validation of a risk score for post-infection irritable bowel syndrome in coastal eastern India.

BACKGROUND AND AIM: Post-infection irritable bowel syndrome (PI-IBS) can occur following acute gastroenteritis (AGE). This study was designed to evaluate the incidence and risk factors of PI-IBS following AGE and to validate a PI-IBS risk score. METHODS: This prospective study was performed between September 2014 and October 2016 on AGE patients by documenting their AGE severity and following up after 3 and 6 months to study the development of IBS (ROME III criteria). The risk score was calculated for all the subjects, and its discrimination ability was tested. RESULTS: Out of 136 hospitalized AGE patients, 35 developed PI-IBS after 6 months. The factors associated with PI-IBS were younger age, longer duration of AGE, anxiety, depression, abdominal pain, bloody stool, vomiting, fever, family history of IBS, and positive stool culture (univariate analysis); however, on multivariate analysis, younger age (adjusted odds ratio [AOR] 0.5; p 0.03), prolonged duration of AGE (AOR 8.6; p 0.01), and abdominal cramps (AOR 2.1; p 0.02) were the independent factors influencing its occurrence. PI-IBS occurred even after infection with Vibrio cholerae. The PI-IBS risk score was significantly higher in patients who developed PI-IBS (72.4 ± 14.48 vs. 31.56 ± 20.4, p-value < 0.001); score > 50 had a sensitivity and specificity of 91.4% and 84.2%, respectively. CONCLUSION: One fourth of AGE patients developed PI-IBS after 6 months. Factors influencing its development were younger age, long duration of AGE, and abdominal pain. The PI-IBS risk score had good predictive accuracy in our population.

Quadruplex PCR for simultaneous detection of serotype, biotype, toxigenic potential, and central regulating factor of Vibrio cholerae.

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.

Incidence of bacterial enteropathogens among hospitalized diarrhea patients from Orissa, India.

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.

Epidemiology and Antibiogram Profile of Vibrio cholerae Isolates between 2004-2013 from Odisha, India.

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.

Prevalence of carrier state theileriosis in lactating cows.

AIM: The objective of this study was to examine the carrier status of theileriosis among apparently healthy cross-bred jersey cattle population of Odisha using conventional blood smear examination and polymerase chain reaction (PCR). MATERIALS AND METHODS: A total of 34 apparently healthy cross-bred Jersey lactating cows were considered in this study. Blood samples were subjected to microscopic examination after staining with Giemsa stain and PCR based molecular diagnosis using two sets of primer, i.e., N516/N517 and TorF1/TorF2 specific for Theileria annulata and Theileriaorientalis, respectively. RESULTS: Examination of blood samples revealed presence of theileria parasites to a magnitude of 20.59% for T. annulata, 8.82% for T. orientalis, and 2.94% for both. CONCLUSION: Molecular diagnosis was found to be much more sensitive than conventional method for diagnosis of theileriosis. T. annulata was found to be the predominant species affecting the exotic cattle. T. orientalis was detected in apparently healthy cows.

Vibrio cholerae O1 Ogawa Strains Carrying the ctxB7 Allele Caused a Large Cholera Outbreak during 2014 in the Tribal Areas of Odisha, India.

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.

Emergence of Vibrio cholerae O1 biotype El Tor serotype Inaba causing outbreaks of cholera in Orissa, India.

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 Ogawa cholera outbreak 6 months after the Inaba cholera outbreaks in India, 2006.

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.