Expression of microRNA-155 correlates positively with the expression of Toll-like receptor 7 and modulates hepatitis B virus via C/EBP-ß in hepatocytes.

Effective recognition of viral infection and successive activation of antiviral innate immune responses are vital for host antiviral defence, which largely depends on multiple regulators, including Toll-like receptors (TLRs) and microRNAs. Several early reports suggest that specific TLR-mediated immune responses can control hepatitis B virus (HBV) replication and express differentially with disease outcome. Considering the versatile function of miR-155 in the TLR-mediated innate immune response, we aimed to study the association between miR-155 and TLRs and their subsequent impact on HBV replication using both a HBV-replicating stable cell line (HepG2.2.15) and HBV-infected liver biopsy and serum samples. Our results showed that miR-155 was suppressed during HBV infection and a subsequent positive correlation of miR-155 with TLR7 activation was noted. Further, ectopic expression of miR-155 in vitro reduced HBV load as evidenced from reduced viral DNA, mRNA and subsequently reduced level of secreted viral antigens (HBsAg and HBeAg). Our results further suggested that CCAAT/enhancer-binding protein-ß (C/EBP-ß), a positive regulator of HBV transcription, was inhibited by miR-155. Taken together, our study established a correlation between miR-155 and TLR7 during HBV infection and also demonstrated in vitro that increased miR-155 level could help to reduce HBV viral load by targeting C/EBP-ß.

Bancroftian filariasis: circulating B-1 cells decreased in microfilaria carriers and correlate with immunoglobulin M levels.

B-1 cells play an important role in the outcome of infection in schistosomiasis, pneumonia and experimental filariasis. However, no information exists regarding status of B-1 cells in clinical manifestations of human filariasis. We investigated the levels of B-1 cells from the total B cells by flow cytometry. Significantly low levels of B-1 cells and IgM antibodies were detected against a wide variety of autoantigens in microfilariae carriers as compared to endemic controls and patients with chronic pathology. A positive correlation was found between IgM antibodies to actin and ss-DNA. Absorption of plasma with soluble actin, myosin and lipopolysaccharides (LPS) resulted in significant removal of antifilarial antibodies. Affinity-purified anti-ss-DNA antibodies were found to be reactive to filarial antigens and various autoantigens. Further, a positive correlation was found between polyreactive antibodies and B-1 cells in filarial-infected human subjects. After antifilarial treatment, levels of IgM antibodies to ss-DNA, actin, LPS and filarial antigen increased significantly indicating a role of polyreactive naturally occurring antibodies in filarial infection. Our findings add to the existing evidence that the B-cell defect in BALB.Xid mice account for susceptibility to murine filarial infection and indicate an important role for these antibodies in providing host protection against filarial infection.

In utero sensitization modulates IgG isotype, IFN-γ and IL-10 responses of neonates in bancroftian filariasis.

In utero exposure has been considered as a risk factor for filarial infection. To evaluate the influence of maternal infection on filarial-specific IgG subclass response in neonates and their correlation with plasma levels IL-10 and interferon-γ, 145 pairs of mothers and their respective cord bloods were examined. Transplacental transfer of circulating filarial antigen (CFA) was observed in 34·8% cord bloods from CFA positive mothers. Filarial-specific IgG1, IgG2 and IgG4 responses of cord bloods were found to be positively correlated with CFA of mothers. In contrast, IgG3 responses negatively correlated with CFA of mothers. The % of similarity of recognition pattern in the cord blood with maternal blood was high for IgG3 response than IgG4 in all three groups. An increased levels of IL-10 and decreased levels of interferon gamma (IFN-γ) were observed in cord blood of infected mothers. Interferon gamma was positively correlated with IgG3 and negatively correlated with IgG4 level. On the other hand, IL-10 was positively correlated with IgG4 and CFA, indicating that cytokines may play a role in modulating the immune responses in cord bloods of sensitized foetus. The findings of the study reveal that in utero tolerance or sensitization may influence the filarial-specific immunity to infection in neonates.

Viral aetiology and clinico-epidemiological features of acute encephalitis syndrome in eastern India.

This study reports clinico-epidemiological features and viral agents causing acute encephalitis syndrome (AES) in the eastern Indian region through hospital-based case enrolment during April 2011 to July 2012. Blood and CSF samples of 526 AES cases were investigated by serology and/or PCR. Viral aetiology was identified in 91 (17·2%) cases. Herpes simplex virus (HSV; types I or II) was most common (16·1%), followed by measles (2·6%), Japanese encephalitis virus (1·5%), dengue virus (0·57%), varicella zoster virus (0·38%) and enteroviruses (0·19%). Rash, paresis and cranial nerve palsies were significantly higher (P < 0·05) with viral AES. Case-fatality rates were 10·9% and 6·2% in AES cases with and without viral aetiology, respectively. Simultaneous infection of HSV I and measles was observed in seven cases. This report provides the first evidence on viral aetiology of AES viruses from eastern India showing dominance of HSV that will be useful in informing the public health system.

Hydrocele fluid: can it be used for immunodiagnosis of lymphatic filariasis?

BACKGROUND & OBJECTIVES: Diagnosis of lymphatic filariasis using serum has been established but the utility of hydrocele fluid for the purpose is not exactly known. Since, hydrocele is a chronic form of the disease manifestation in a variety of situations and often poses difficulty in diagnosing its origin, we have evaluated the usefulness usage of hydrocele fluid for diagnosis of filarial origin of hydrocele in this study. METHODS: Paired samples of serum and hydrocele fluid from 51 individuals with hydrocele, living in an endemic area of Wuchereria bancrofti were assessed. Circulating filarial antigen, filarial specific antibody and cytokine assay were performed in both serum and hydrocele fluid of patients. RESULTS: Og4C3 assay detected circulating filarial antigen (CFA) in serum and corresponding hydrocele fluids. The level of IgG, IFN-γ and IL-10 were found to be high in CFA-negative, while IgM and IgE were high in CFApositive hydrocele fluid and serum samples associated with hydrocele. On the other hand neither CFA-positive nor CFA-negative hydrocele fluid and serum samples associated with hydrocele showed any difference in IgG4 level. INTERPRETATION & CONCLUSION: This study showed that the filaria related antigens and antibodies found in serum can be detected with equal sensitivity in hydrocele fluid. Therefore, it can be used as an alternative to serum for immunodiagnosis of filariasis, and help monitoring the filarisis elimination programme.

Immuno-epidemiology of bancroftian filariasis: a 14-year follow-up study in Odisha, India.

Forty asymptomatic, circulating filarial antigen negative (CFA(-ve)) and ten asymptomatic, circulating filarial antigen positive (CFA(+ve)) individuals were followed up longitudinally over a period of 14 years at intervals of 7 years in order to investigate the immunological, parasitological and clinical changes that took place in an endemic area due to natural process. The clinical status, microfilaremia, circulating filarial antigenemia and immunological responses to filarial antigens (DSSd1 and Sd30) prepared from cattle filarial parasite Setaria digitata, were examined. The observations showed that 19 individuals had developed either antigenemia or filarial symptoms (acute filarial lymphangitis/hydrocele) from CFA(-ve) group. Three individuals had cleared antigenemia and one had developed microfilaremia from CFA(+ve) group after 7 years. Increased IgG and IgM and low IgG2 and IgG4 level responses along with high lymphocyte production were observed in CFA-negative individuals. This was in contrast to observations made in CFA(+ve) subjects. The results of the present study indicated that the changes taking place in the immunological, clinical and CFA status of individuals residing in filaria endemic regions developed different clinical manifestation with course of time.

Rapid spread of Vibrio cholerae O1 El Tor variant in Odisha, Eastern India, in 2008 and 2009.

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.

Large outbreak of cholera caused by El Tor variant Vibrio cholerae O1 in the eastern coast of Odisha, India during 2009.

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.

Maternal filarial infection: association of anti-sheath antibody responses with plasma levels of IFN-γ and IL-10.

Maternal filarial infection influences the risk of acquiring infection and development of immunity in children. Here we have analysed the blood samples of 60 mothers (24 infected and 36 uninfected) and their corresponding cord bloods to assess the impact of maternal infection on the anti-sheath antibodies and cytokine production in neonates born from them. About 69·4% of non-infected mothers and their cord bloods showed the presence of anti-sheath antibodies, while only 16·6% of the cord bloods from infected mothers were positive for it. The IL-10 level was significantly high in cord bloods of infected mothers compared with non-infected mothers. At the same time the IL-10 level was also observed to be remarkably high in cord bloods of both infected and non-infected mothers negative for anti-sheath antibody. In contrast, IFN-γ levels were significantly high in cord bloods of non-infected mothers compared with infected mothers and the increment was prominent in cord bloods of both infected and non-infected mothers positive for anti-sheath antibody. The study reveals that the presence or absence of anti-sheath antibodies in association with cytokines skews the filarial specific immunity to either Th1 or Th2 responses in neonates. This may affect the natural history of filarial infection in early childhood.

A multimodal detection model of dolphins to estimate abundance validated by field experiments.

Abundance estimation of marine mammals requires matching of detection of an animal or a group of animal by two independent means. A multimodal detection model using visual and acoustic cues (surfacing and phonation) that enables abundance estimation of dolphins is proposed. The method does not require a specific time window to match the cues of both means for applying mark-recapture method. The proposed model was evaluated using data obtained in field observations of Ganges River dolphins and Irrawaddy dolphins, as examples of dispersed and condensed distributions of animals, respectively. The acoustic detection probability was approximately 80%, 20% higher than that of visual detection for both species, regardless of the distribution of the animals in present study sites. The abundance estimates of Ganges River dolphins and Irrawaddy dolphins fairly agreed with the numbers reported in previous monitoring studies. The single animal detection probability was smaller than that of larger cluster size, as predicted by the model and confirmed by field data. However, dense groups of Irrawaddy dolphins showed difference in cluster sizes observed by visual and acoustic methods. Lower detection probability of single clusters of this species seemed to be caused by the clumped distribution of this species.

Host genotype affects endotoxin release in excreta of broilers at slaughter age.

Host genotype, early post-hatch feeding, and pre- and probiotics are factors known to modulate the gut microbiome. However, there is a knowledge gap on the effect of both chicken genotype and these dietary strategies and their interplay on fecal microbiome composition and diversity, which, in turn, can affect the release of endotoxins in the excreta of broilers. Endotoxins are a major concern as they can be harmful to both animal and human health. The main goal of the current study was to investigate whether it was possible to modulate the fecal microbiome, thereby reducing endotoxin concentrations in the excreta of broiler chickens. An experiment was carried out with a 2 × 2 × 2 factorial arrangement including the following three factors: 1) genetic strain (fast-growing Ross 308 vs. slower growing Hubbard JA757); 2) no vs. combined use of probiotics and prebiotics in the diet and drinking water; and 3) early feeding at the hatchery vs. non-early feeding. A total of 624 Ross 308 and 624 Hubbard JA757 day-old male broiler chickens were included until d 37 and d 51 of age, respectively. Broilers (N = 26 chicks/pen) were housed in a total of 48 pens, and there were six replicate pens/treatment groups. Pooled cloacal swabs (N = 10 chickens/pen) for microbiome and endotoxin analyses were collected at a target body weight (BW) of 200 g, 1 kg, and 2.5 kg. Endotoxin concentration significantly increased with age (p = 0.01). At a target BW of 2.5 kg, Ross 308 chickens produced a considerably higher amount of endotoxins (Δ = 552.5 EU/mL) than the Hubbard JA757 chickens (p < 0.01). A significant difference in the Shannon index was observed for the interaction between the use of prebiotics and probiotics, and host genotype (p = 0.02), where Ross 308 chickens with pre-/probiotics had lower diversity than Hubbard JA757 chickens with pre-/probiotics. Early feeding did not affect both the fecal microbiome and endotoxin release. Overall, the results suggest that the chicken genetic strain may be an important factor to take into account regarding fecal endotoxin release, although this needs to be further investigated under commercial conditions.

Effect of chlorine dioxide and sodium hypochlorite on the dissolution of human pulp tissue - An in vitro study.

BACKGROUND: Organic tissue dissolution is an important property of an irrigant which aids in the success of root canal treatment. Recent studies have advocated the use of Chlorine dioxide as an endodontic irrigant. The aim of this study is to compare the dissolution efficacy of chlorine dioxide and sodium hypochlorite on human pulp tissue. METHODS: In this study, 2% Sodium hypochlorite, 5% Chlorine dioxide and isotonic saline solution (control) were used. Thirty human pulp tissue specimens were exposed to three test solutions (n = 10) for 30 min following which the loss of weight was compared from the original weight by using a digital analytical balance. RESULTS: Sodium hypochlorite was more efficient in dissolving human pulp tissue when compared to Chlorine dioxide. Isotonic saline solution failed to dissolve any of the specimens. CONCLUSION: 5% Chlorine dioxide is capable of dissolving human pulp tissue but sodium hypochlorite was more effective.

Enriching table eggs with n-3 polyunsaturated fatty acids through dietary supplementation with the phototrophically grown green algae Nannochloropsis limnetica: effects of microalgae on nutrient retention, performance, egg characteristics and health parameters.

The fatty acid content of microalgae, especially the high content of omega-3 fatty acids such as eicosapentaenoic acid (EPA, C20:5) and docosahexaenoic acid (DHA, C22:6), could enrich eggs when fed to laying hens. Moreover, the properties and bioactive components of omega-3 fatty acids could positively influence the health and production performance of laying hens. In this study, the effects of dried Nannochloropsis limnetica inclusions in diets on yolk omega-3 fatty acid content, laying hen performance, nutrient retention, intestinal morphometry and systemic inflammatory markers were measured. A total of 240 twenty-five-wk-old laying hens were randomly assigned to 5 treatments distributed among 30 pens. Treatment A received the reference diet, while diets in treatments B, C, and D contained the control diet with 1, 2, and 3% microalgae added, respectively. In treatment E, a portion of ingredients of the control diet was replaced with rapeseed meal to induce a mild nutritional challenge, along with an inclusion of 3% microalgae. Compared to the control group the rate of lay increased by approximately 5% (P = 0.039) when birds were fed 2 or 3% microalgae. Furthermore, inclusion of 2 and 3% microalgae resulted in higher feed intake compared to the control group (126, 125, and 119 g/hen/d respectively; P = 0.001). Other performance parameters such as nutrient retention and egg characteristics were not affected by the dietary treatments. The EPA and DHA content of the yolk increased with increasing microalgae inclusion level (P < 0.001). A 2% algal inclusion resulted in 58.3 (EPA) and 603 (DHA) mg per 100 g dry yolk, respectively. Plasma haptoglobin levels of laying hens in both treatments receiving 3% microalgae were almost 3 times lower compared to the control group (1.25 and 1.62 vs. 5.60; P < 0.001), regardless of the inclusion of rapeseed in the diet. Based on these results, it can be concluded that the inclusion of N. limnetica enriches the egg yolk without negatively affecting the performance of laying hens and egg characteristics. Due to the positive effect on feed intake, microalgae in the diet provide nutritional benefits for laying hens. However, the positive effects of microalgae, especially on the health of laying hens, warrants further research.

Rapid spread of chikungunya virus infection in Orissa: India.

BACKGROUND & OBJECTIVES: A large number of cases of undiagnosed fever and joint pain were reported from different parts of the State of Orissa since February 2006. Epidemiological and laboratory investigation were carried out to confirm the cause of emerging illness, which was provisionally suspected as Chikungunya (CHIK) fever. METHODS: Upon getting the reports of suspected CHIK like illness in different parts of the State, epidemic investigations were carried out in the outbreak affected villages. Case history was recorded, clinical examination undertaken and blood samples collected for seroconfirmation for CHIK IgM antibody using ELISA based kit. Simultaneously vector survey was also carried out. RESULTS: With no previous record of CHIK infection in the State, the first outbreak was confirmed during February 2006. Subsequently, the infection spread to 13 of 30 districts in different episodes covering 79 villages till November 2007. Attack rate was 9-43 per cent in the different outbreaks with average seropositivity of 24 per cent to CHIK specific IgM. Morbidity was high though no deaths were recorded. Aedes aegypti and Ae. albopictus were identified as the possible vectors for transmission. INTERPRETATION & CONCLUSIONS: The report confirmed emergence of CHIK infection in the State of Orissa, India, and its spread to a larger geographic zone in a short period which warrants public health measures to control further spread.

Transplacental transfer of filarial antigens from Wuchereria bancrofti-infected mothers to their offspring.

OBJECTIVE: Maternal infection has been considered to be a risk factor for filarial infection in offspring. In order to examine the influence of maternal infection in neonates, we have determined the prevalence of circulating filarial antigen (CFA) and anti-filarial antibodies in 119 maternal and corresponding cord blood samples collected from an area endemic for bancroftian filariasis. METHOD: Prevalence of antigenaemia was detected using Og4C3 circulating filarial antigen enzyme-linked immunosorbent assay. The presence of microfilariae was determined by filtration of a 1 ml sample through a Nuclepore membrane. Antibody isotypes (IgG, IgM, and IgE) to filarial antigen (Setaria digitata antigenic extract) were determined by enzyme linked immunosorbent assay (ELISA). RESULTS: Microfilariae were detected in 14 cases (11.8%), whereas the Og4C3 assay could detect filarial antigen in 44.5% of pregnant mothers. Interestingly, 24.5% of samples born from CFA-positive mothers were found positive for CFA. None of the cord samples from CFA-negative mothers were found positive for CFA. No significant difference was observed in prevalence of filarial-specific IgG, IgM and IgE antibodies in CFA-positive and negative mothers. IgG antibody was detected in 60.5% of maternal and 21.8% of cord samples. IgG antibody in the cord does not differ with the antigen status of the mother. In contrast IgM and IgE antibody prevalence was significantly higher in cord from infected mothers than non-infected mothers (11.3% vs 0 for IgM, 24.5% vs 3.03% for IgE). CONCLUSION: Our study demonstrates the transplacental transfer of circulating filarial antigen from mother to cord. Filaria-specific IgM and IgE antibodies were higher in cord blood from infected mothers than from non-infected mothers. The findings of the study provide additional circumstantial evidence for pre-natal sensitization to filarial antigens developed in utero.

Domestic violence in Eastern India: factors associated with victimization and perpetration.

OBJECTIVES: To examine the factors associated with victimization and perpetration of domestic violence in Eastern India. STUDY DESIGN: Population-based cross-sectional study. METHODS: Married women (n=1718) and men (n=1715) from three Eastern Indian states were included in this study. Univariate and multiple logistic regression models were used to identify the determinants of domestic violence. RESULTS: Age, education, occupation, marital duration and husband's alcoholism emerged as significant predictors of victimization and perpetration of all types of domestic violence. A higher level of family income was found to be highly protective against the risk of violence. In addition, other risk and protective factors for victimization and perpetration of each type of violence were identified. CONCLUSIONS: This study contributes to the violence literature by shedding light on the risk factors of perpetration and victimization of domestic violence. These results provide vital information to develop interventions, as well as policies and programmes towards preventing violence. Also, this knowledge facilitates healthcare personnel to intervene more effectively with women at risk of violence-related health problems.

The CCTTT pentanucleotide microsatellite in iNOS promoter influences the clinical outcome in P. falciparum infection.

To assess the hypothesis that nitric oxide (NO) is critical in the pathogenesis of cerebral malaria, we analyzed those single nucleotide polymorphisms (SNPs) and microsatellite (MS) of the promoter region of inducible nitric oxide synthase (iNOS) gene which are known to enhance the NO production in vivo. A total of 428 (204 severe, 224 mild) adult patients living in the eastern part of India were analyzed. The single nucleotide substitutions -954G-->C was found to be very rare, and -1173C-->T was absent in this population. But interestingly, longer forms of MS were found to be significantly associated with severe malaria (OR = 2.89, 95% CI = 1.955-4.295, P < 0.0001), and the linear regression analysis revealed that the risk of severe malaria significantly increases as the summed repeat number in an individual increase (OR = 1.16, P = 0.0013). Further, the median plasma level of nitrate/nitrite (NOx) was observed to be high in mild patients compared to severe patients, and the level of parasitemia was significantly low among mild patients than severe ones. These findings suggest that the CCTTT repeats in iNOS may play a key role in the pathogenesis of severe malaria.

An outbreak of cholera associated with an unprotected well in Parbatia, Orissa, Eastern India.

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.

An Investigation on the Coinfection of Measles and HSV-1 in Hospitalized Acute Encephalitis Syndrome Patients in Eastern India.

Acute encephalitis syndrome (AES) is a clinical condition that occurs due to infectious and noninfectious agents- however, viruses are considered to be the dominant pathogen. agents- however, viruses are considered to be the dominant pathogen. In this study, suspected AES cases were enrolled and tested for viral etiology through serology and polymerase chain reaction (PCR)/reverse transcriptase PCR from August 2012-July 2013. During this period, 820 cases were investigated and 96 cases were diagnosed to have a viral etiology whereas 20 patients had IgM antibodies for measles in serum and HSV-1 DNA in cerebrospinal fluid. All 20 of the patients were children below 14 years of age. The median hospital stay was 15 days (IQR: 14.2-17 days) and median GCS score was 7(IQR: 6-8) and were significantly different with patients with co-infections when comapred with patients having HSV-1 infection only. It may be suspected that the measles infection may have a role in the pathogenesis and thus an impact on the prognosis of the AES when present with HSV-1.

Molecular and phylogenetic analysis of the dengue strains circulating in Odisha, India.

Dengue has emerged as a major public health challenge in terms of both changing clinical pattern and epidemiological features. The state of Odisha reported first dengue epidemic in the year 2010 and this continued each year in epidemic form during post monsoon period gradually becoming an endemic phenomenon. Present study depicts the changing epidemiological and clinical pattern of dengue with reference to its serotypes and genotypes. The study included 5320 suspected dengue cases from different health facilities of the state during 2010-2017. Dengue NS1 antigen and IgM antibody was done through ELISA. Serotyping was done through RTPCR by amplifying a part of core-pre-membrane gene (CprM) followed by sequencing and phylogenetic analysis. Dengue IgM antibody in 17.7% cases and NS1 antigen in 53.20% cases was detected. Dengue serotype 2 (DEN-2) was the only serotype detected in 2010 and 2011 where as all four serotypes 1, 2, 3, 4 were detected in 2012-2017, DEN-2 being dominant but in 2017 DEN-3 was found to be dominant. Phylogenetic analysis revealed genotype IV of DEN-2 and genotype III of DEN-1 and DEN-3 circulating in this region. In 6 cases involvement of DEN-2 in clinically evident encephalitis cases is an important observation in this region and needs public health attention. High prevalence of dengue was observed without any previous reported outbreaks in the state with increased number of cases from 2010 to 2012 affecting both urban and rural areas. High incidence in 2012 was due to co-circulation of more than one serotype which continued in the following years. Severity in some cases was associated with mixed infection but in most cases it was mild indicating the endemic nature of the virus in most parts of Odisha.