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.

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.

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.

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.

Epidemiological and clinical profile of Influenza A(H1N1) pdm09 in Odisha, eastern India.

Epidemic of flu is highly contagious and it spreads through air. In 2009 H1N1 influenza virus emerged after reassortment of North American TRIG and Eurasia Avian like virus of swine and started epidemic in Mexico. The first cases were reported from Hyderabad city on 16th May 2009 in India that spread rapidly within a short span of time. During this period large population of Odisha situated at the eastern side of India was also affected and incidences of H1N1 cases were recorded through state Government surveillance system. In this study real time RT-PCR based diagnosis was conducted for the throat swabs collected from suspected H1N1 cases in Odisha during 2009-2017. A total of 2872 throat swabs were received from 23 different Government and private hospitals and 21.1% positivity was confirmed. The disease affected mostly 46-60 years age group, males (50.6%) being more affected. The clinical features had shown that fever with cough (89.6%) was the most common symptom followed by shortness of breath (72.7%). Post monsoon was the peak season in which most of the cases were reported. Neurological signs, pregnancy, diabetes and hypertension were found to be risk factors for H1N1. The case fatality rate (CFR) was 15%.

Occult Hepatitis B virus Infection (OBI) predominated by genotype D among chronic liver disease patients: The first report from Odisha, Eastern India.

Detection of Occult hepatitis B infection (OBI) has higher significance for treatment and management. There is no information available about the OBI and its genotype associated with chronic hepatitis patients in Odisha, India. We aimed to determine the association of OBI and its genotype among the chronic hepatitis patients in Odisha. In a hospital based study, 175 serum samples of chronic hepatitis patients were screened for Hepatitis surface antigen (HBsAg). All the HBsAg - negative samples were tested for hepatitis B core antibodies (HBcAb). HBcAb-positive samples were tested further for Hepatitis surface antibodies (HBsAb), hepatitis B virus HBV-DNA and HBV genotyping. Of the 89 HBsAg negative samples, 79 (88.8%) were found positive for HBcAb and these patients were presumed to have OBI. Among the total HBcAb positive samples, 22 (27.8%) were positive and 57 (72.2%) were negative for HBsAb. Detection of HBV- DNA from 45 out of 79 HBcAb positive samples (57.0%) yielded positive results for OBI which was negative for HBsAb. Genotyping of all hepatitis B virus showed that all 45 (100%) were genotype-D. Detection of OBI among chronic hepatitis patients suggests testing for OBI for management and treatment.

Cholera prevention and control in Asian countries.

Cholera remains a major public health problem in many countries. Poor sanitation and inappropriate clean water supply, insufficient health literacy and community mobilization, absence of national plans and cross-border collaborations are major factors impeding optimal control of cholera in endemic countries. In March 2017, a group of experts from 10 Asian cholera-prone countries that belong to the Initiative against Diarrheal and Enteric Diseases in Africa and Asia (IDEA), together with representatives from the World Health Organization, the US National Institutes of Health, International Vaccine Institute, Agence de médecine préventive, NGOs (Save the Children) and UNICEF, met in Hanoi (Vietnam) to share progress in terms of prevention and control interventions on water, sanitation and hygiene (WASH), surveillance and oral cholera vaccine use. This paper reports on the country situation, gaps identified in terms of cholera prevention and control and strategic interventions to bridge these gaps.

Epidemiological features and genetic characterization of virus strains in rotavirus associated gastroenteritis in children of Odisha in Eastern India.

We have studied the clinical characteristics, severity and seasonality of rotavirus infection and prevalent genotypes in 652 non-rota vaccinated children in Odisha in eastern India. P genotypes were analysed for their association with host blood group antigens. P type of the virus is determined by the VP8* gene, and specific recognition of A - type of Histo - blood group antigen by P[14]VP8* has been reported. VP4, VP7 and VP6 genes of commonly identified G1P[8] strain were compared with genes of the same strain isolated from other parts of India, elsewhere and strains used for Rotarix and Rotateq vaccines. In 54.75% of children with gastroenteritis, rota virus was found. 9.65% of children had moderate, 78.07% severe, and 12.28% very severe disease as assessed using the Vesikari scoring system. The incidence of infection was highest during winter months. There was no association between any blood group and specific P genotypes. G1P[8] was the commonest cause of gastroenteritis, followed by G1P[11], G3P[8], G9P[8], G2P[4], G2P[6], G9P[4], G9P[11] and G1P[6]. Predominant G genotypes identified were G1 (72.9%), G9 (10.81%), G2 (8.10%) and G3 (8.10%). Sequence analysis of the VP7 gene, placed the G1P[8] strain in lineage 1 and of VP6 gene placed nine G1P[8] strains in subgroup II and one in subgroup I. The VP7 gene segment of two Odisha G1P[8] strains were found to cluster relatively close to the VP7 sequences of Rotarix vaccine. Antigenic differences were found with vaccine strains. Ten G1P[8] strains sequenced for the VP4 gene had 91-93% nucleotide and 92-96% amino acid identity with Rotateq vaccine P[8]). Rotarix vaccine VP4 had 89-91% nucleotide and 90-92% amino acid identity. Our findings indicate genetic variability of rotavirus strains circulating in the region and are significant, given the introduction of rota vaccination in the State.

Association of Rotavirus Gastroenteritis with Histo-blood Group Antigens.

Association of rotavirus gastroenteritis with histo-blood group antigens in children younger than 5 years admitted with diarrhea (n=389) was studied. Distribution of blood groups in rotavirus positive (n=96) and rotavirus negative (n=51) diarrhea gastroenteritis cases did not show any susceptibility to any blood group; blood group O seemed to be protective.

Feeding stage, species, body part and sex-specific activity of glutathione S-transferase in mosquito.

In the present study, the feeding stage, body parts, development and sex specific activity of Glutathione S-transferases (GSTs) were observed in different mosquito species (Aedes aegypti, Culex quinquefasciatus, Anopheles stephensi, An. culicifacies, An. annularis, An. subpictus, An. vagus). GST activity was assayed spectrophotometrically at 23°C, using a UV Max microplate Reader, to measure the rate of conjugation of GSH to CDNB. A significant species-specific difference in the activity of GST was noticed, highest being in unfed Ae. aegypti (41.2 nmol/min/mg) followed by unfed Cx. quinquefasciatus (7.9 nmol/min/mg) and the least in unfed An. stephensi (5.8 nmol/min/mg). In all the species the GST activity was found to be significantly higher in fully fed and gravid stages compared with the unfed, while the enzyme activity was reduced after egg laying either to the level of unfed animals or well below its level in all the experimental species. The GST activity was found to be higher in the abdominal region of all the experimental species in comparison with the other body parts (head and thorax). The GST activity of An. stephensi increased gradually through the larval stages and reached the maximum level in the pupae and remained at that level in the newly emerged adults. However, its activity declined markedly (10 fold) with ageing from 5 to 40 days. A significant sex-related difference in the specific activity of GST was found in An. stephensi where approximately 3.5 fold lower activity was observed in males compared with its females, whereas no significant variation was noticed in Ae. aegypti and Cx. quinquefasciatus. The study corroborates the fact that GSTs are differentially regulated by multiple mechanisms in response to xenobiotics modulation in situation-specific manner such as species, sex, feeding and developmental stage. The knowledge of situation-specific modulation of GST will provide a better understanding of GST based insecticide resistance mechanism which is essential for the design of sensitive monitoring methods and for an effective insecticide resistance management. The findings are significant in terms of the methods used to control mosquito vector population.

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.

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.

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.

Spot or early morning sample for mycobacterial culture: which?

Drug resistance studies ideally require two sputum specimens, one spot and one early morning, for culture in Löwenstein-Jensen medium. Although the type of specimen plays a major role in the detection of acid-fast bacilli using microscopy, this type of specimen collection puts unnecessary pressure on patients. Two sputum samples, one on the spot and another in the early morning, collected from 179 TB patients in cetylpyridinium chloride and processed for culture, yielded respectively 89.9% and 87.7% culture positivity, while culture positivity increased to 93.3% when both specimens were tested.

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.

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.

Low levels of anti TB drug resistance in Rayagada district of Odisha, India.

A study was conducted at Rayagada district of Odisha, India, among smear-positive tuberculosis (TB) patients to determine the resistance pattern to first-line drugs. Sputum samples were collected from 405 new and 37 previously treated patients and were tested at Regional Medical Research Centre, Bhubaneswar. Resistance to any anti-tubercular drug was observed to be 5.2% among new cases and 16.1% among previously treated patients, while multidrug-resistant tuberculosis (MDR-TB) was found to be 0% in new and 8.1% in previously treated cases. Such a low level of resistance may be due to the limited use of TB drugs outside the ongoing program.

Decontamination of sputum for longer time in sodium hydroxide for isolation of Mycobacterium tuberculosis.

Decontamination by modified Petroff's method is being practiced in many laboratories carrying out Mycobacterium tuberculosis culture and drug susceptibility testing. The method exposes mycobacteria to 4% sodium hydroxide for 30min. However, laboratories in developing countries with limited resources might be using a type of centrifuge that does not open during power failures and exposes the mycobacteria to alkali for longer periods. Out of 28 smear-positive specimens processed, 85.7%, 85.7% and 60.7% of specimens showed a positive culture after exposure to alkali for 0.5, 1.0 and 72h. Laboratories compelled to expose the mycobacteria for a longer duration of time can still attempt isolation for culture as only a small amount of bacteria are needed for culture positivity.

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.