Fecal Virus Research Articles

21 MyD88 Is Important for Rotavirus Infection Containment and Specific Adaptive Responses

Background: Rotavirus (RV), the world's leading cause of severe viral gastroenteritis amongst children, primarily infects small intestinal epithelial cells (IECs) and is responsible for over 500,000 deaths and millions of physician visits/hospitalizations per year. While successful resolution of infection and protection against future infection is mediated by adaptive immunity, particularly mucosal IgA, relatively little is known how innate immunity contains acute infection and drives the adaptive immune response to this virus. In light of the broad role of MyD88 in signaling by TLRs and inflammasome cytokine receptors signaling, our goal was examine the consequence of absence of MyD88 in a mouse model of RV infection as means to mechanistically explore innate immunity to this virus. Methods: Suckling and adult mice (WT, MyD88KO, Caspase-1KO, and various bone-marrow irradiation chimeras) mice were inoculated with mouse-passaged RVEC strain. Infection was assessed by ELISA and qRT-PCR. RV replication was assessed by quantitating + to strand ratios. Diarrhea was scored visually. RV-specific immune responses were measured by fecal/serum ELISA. Results: Loss of MyD88 markedly impaired innate immunity to RV resulting in higher levels of virus in feces, intestinal lysates, and peripheral blood cells as well as increased RV replicative ability. Such increased RV levels correlated with a marked increase in the incidence and duration of diarrhea in neonatal mice. These results were fully phenocopied by loss of Caspase-1 and partially mimicked by loss of IL-1 and IL-18 suggesting a role for inflammasome signaling in innate control of RV infection. Such inability to control RV was not observed in irradiated WTmice reconstituted with MyD88KO or Caspase-1KO bone marrow nor could WT bone marrow restore innate immunity to RV in MyD88KO or Caspase-1KO mice suggesting a prominent role for inflammasome signaling in IEC in limiting RV infection/ pathogenesis. Loss of MyD88 also profoundly impaired adaptive immunity to RV. Specifically, MyD88KOmice produced less serum RV-specific IgG and IgA and have Th2 skewed antibody responses indicative of improper Th responses and isotype class switching. In contrast to the role of MyD88 in innate immunity, MyD88-mediated adaptive immunity was independent of Caspase-1, IL-1, and IL-18 and was mediated by MyD88 in bone marrow derived cells. Conclusion: Inflammasome signaling, in IECs, plays a major role in limiting RV infection and, consequently, the diarrheal disease that the great mortality/morbidity caused by this pathogen. TLR signaling in bone-marrow derived cells is important for driving the humoral immune responses that provide lasting protection against this pathogen.

Identification of a novel single-stranded circular DNA virus in pig feces.

Porcine stool-associated circular virus 5 (PoSCV5) was detected in the feces of a pig with diarrhea. The complete 3,062-nucleotide genome contains two bidirectionally transcribed open reading frames (ORFs). Phylogenetic analysis of the deduced replication initiator protein (Rep) places PoSCV5 alone on a deep branch among the small circular Rep-encoding single-stranded DNA viruses.

A probabilistic model of gastroenteritis risks associated with consumption of street food salads in Kumasi, Ghana: Evaluation of methods to estimate pathogen dose from water, produce or food quality

With a rapidly growing urban population in Kumasi, Ghana, the consumption of street food is increasing. Raw salads, which often accompany street food dishes, are typically composed of perishable vegetables that are grown in close proximity to the city using poor quality water for irrigation. This study assessed the risk of gastroenteritis illness (caused by rotavirus, norovirus and Ascaris lumbricoides) associated with the consumption of street food salads using Quantitative Microbial Risk Assessment (QMRA). Three different risk assessment models were constructed, based on availability of microbial concentrations: 1) Water — starting from irrigation water quality, 2) Produce — starting from the quality of produce at market, and 3) Street — using microbial quality of street food salad. In the absence of viral concentrations, published ratios between faecal coliforms and viruses were used to estimate the quality of water, produce and salad, and annual disease burdens were determined. Rotavirus dominated the estimates of annual disease burden (~10−3Disability Adjusted Life Years per person per year (DALYs pppy)), although norovirus also exceeded the 10−4DALY threshold for both Produce and Street models. The Water model ignored other on-farm and post-harvest sources of contamination and consistently produced lower estimates of risk; it likely underestimates disease burden and therefore is not recommended. Required log reductions of up to 5.3 (95th percentile) for rotavirus were estimated for the Street model, demonstrating that significant interventions are required to protect the health and safety of street food consumers in Kumasi. Estimates of virus concentrations were a significant source of model uncertainty and more data on pathogen concentrations is needed to refine QMRA estimates of disease burden.

Anti-rotavirus effects by combination therapy of stevioside and Sophora flavescens extract

Anti-rotaviral activities of Sophora flavescens extract (SFE) and stevioside (SV) from Stevia rebaudiana Bertoni either singly or in various combinations were examined in vitro and in vivo using a porcine rotavirus G5[P7] strain. Combination of SFE and SV inhibited in vitro virus replication more efficiently than each single treatment. In the piglet model, SV had no effect on rotavirus enteritis, whereas SFE improved but did not completely cure rotaviral enteritis. Interestingly, combination therapy of SFE and SV alleviated diarrhea, and markedly improved small intestinal lesion score and fecal virus shedding. Acute toxicity tests including the piglet lethal dose 50, and body weight, organ weight and pathological changes for the combination therapy did not show any adverse effect on the piglets. These preliminary data suggest that the combination therapy of SV and SFE is a potential curative medication for rotaviral diarrhea in pigs. Determination of the efficacy of this combination therapy in other species including humans needs to be addressed in the future.

Easy-to-use rapid gene amplification method for direct detection of RNA and DNA viruses in sera and feces from various animals

The development of rapid and simple gene amplification tests is required for detection of pathogens to prevent transmission of infectious diseases between animals or from animals to humans. An easy-to-use rapid gene amplification method that can directly detect RNA and DNA viruses in clinical samples was developed. This method is based on combining loop-mediated isothermal amplification (LAMP) or reverse transcription-LAMP (RT-LAMP) and RNA GEM Tissue, a thermophilic enzyme that extracts nucleic acid by quickly digesting proteins and ribonucleases. The authors named these methods GEM LAMP and GEM RT-LAMP. These methods were able to detect viral DNA and RNA within 70 min in a single tube using only a water bath. The detection capacities were 10–100-fold more sensitive than those of previously established LAMP and RT-LAMP methods. The GEM LAMP and GEM RT-LAMP methods were used to detect macroscopically the presence of DNA and RNA viruses in sera or fecal samples from cattle, pigs, horses, dolphins, penguins, and sea lions using SYBR green I. The GEM LAMP and GEM RT-LAMP methods thus have considerable versatility as tools for detecting pathogens and are applicable to basic human and veterinary medicine, environmental hygiene, and point-of-care-testing.

Management of Hepatitis E Virus (HEV) Zoonotic Transmission: Protection of Rabbits against HEV Challenge following Immunization with HEV 239 Vaccine

Hepatitis E virus (HEV) constitutes a significant health burden worldwide, with an estimated approximately 33% of the world’s population exposed to the pathogen. The recent licensed HEV 239 vaccine in China showed excellent protective efficacy against HEV of genotypes 1 and 4 in the general population and pregnant women. Because hepatitis E is a zoonosis, it is also necessary to ascertain whether this vaccine can serve to manage animal sources of human HEV infection. To test the efficacy of the HEV 239 vaccine in protecting animal reservoirs of HEV against HEV infection, twelve specific-pathogen-free (SPF) rabbits were divided randomly into two groups of 6 animals and inoculated intramuscularly with HEV 239 and placebo (PBS). All animals were challenged intravenously with swine HEV of genotype 4 or rabbit HEV seven weeks after the initial immunization. The course of infection was monitored for 10 weeks by serum ALT levels, duration of viremia and fecal virus excretion and HEV antibody responses. All rabbits immunized with HEV 239 developed high titers of anti-HEV and no signs of HEV infection were observed throughout the experiment, while rabbits inoculated with PBS developed viral hepatitis following challenge, with liver enzyme elevations, viremia, and fecal virus shedding. Our data indicated that the HEV 239 vaccine is highly immunogenic for rabbits and that it can completely protect rabbits against homologous and heterologous HEV infections. These findings could facilitate the prevention of food-borne sporadic HEV infection in both developing and industrialized countries.

Bacterial lipopolysaccharide binding enhances virion stability and promotes environmental fitness of an enteric virus.

Enteric viruses, including poliovirus and reovirus, encounter a vast microbial community in the mammalian gastrointestinal tract, which has been shown to promote virus replication and pathogenesis. Investigating the underlying mechanisms, we find that poliovirus binds bacterial surface polysaccharides, which enhances virion stability and cell attachment by increasing binding to the viral receptor. Additionally, we identified a poliovirus mutant, VP1-T99K, with reduced lipopolysaccharide (LPS) binding. Although T99K and WT poliovirus cell attachment, replication, and pathogenesis in mice are equivalent, VP1-T99K poliovirus was unstable in feces following peroral inoculation of mice. Consequently, the ratio of mutant virus in feces is reduced following additional cycles of infection in mice. Thus, the mutant virus incurs a fitness cost when environmental stability is a factor. These data suggest that poliovirus binds bacterial surface polysaccharides, enhancing cell attachment and environmental stability, potentially promoting transmission to a new host.

The interaction of human microbial pathogens, particulate material and nutrients in estuarine environments and their impacts on recreational and shellfish waters.

Anthropogenic activities have increased the load of faecal bacteria, pathogenic viruses and nutrients in rivers, estuaries and coastal areas through point and diffuse sources such as sewage discharges and agricultural runoff. These areas are used by humans for both commercial and recreational activities and are therefore protected by a range of European Directives. If water quality declines in these zones, significant economic losses can occur. Identifying the sources of pollution, however, is notoriously difficult due to the ephemeral nature of discharges, their diffuse source, and uncertainties associated with transport and transformation of the pollutants through the freshwater-marine interface. Further, significant interaction between nutrients, microorganisms and particulates can occur in the water column making prediction of the fate and potential infectivity of human pathogenic organisms difficult to ascertain. This interaction is most prevalent in estuarine environments due to the formation of flocs (suspended sediment) at the marine-freshwater interface. A range of physical, chemical and biological processes can induce the co-flocculation of microorganisms, organic matter and mineral particles resulting in pathogenic organisms becoming potentially protected from a range of biotic (e.g. predation) and abiotic stresses (e.g. UV, salinity). These flocs contain and retain macro- and micro- nutrients allowing the potential survival, growth and transfer of pathogenic organisms to commercially sensitive areas (e.g. beaches, shellfish harvesting waters). The flocs can either be transported directly to the coastal environment or can become deposited in the estuary forming cohesive sediments where pathogens can survive for long periods. Especially in response to storms, these sediments can be subsequently remobilised releasing pulses of potential pathogenic organisms back into the water column leading to contamination of marine waters long after the initial contamination event occurred. Further work, however, is still required to understand and predict the potential human infectivity of pathogenic organisms alongside the better design of early warning systems and surveillance measures for risk assessment purposes.

Prenatal vitamin A deficiency impairs adaptive immune responses to pentavalent rotavirus vaccine (RotaTeq®) in a neonatal gnotobiotic pig model

Vitamin A deficiency (VAD) is associated with increased childhood mortality and morbidity in impoverished Asian and African countries, but the impact of VAD on rotavirus (RV) vaccine or infection is poorly understood. We assessed effects of gestational and dietary induced pre- and post-natal VAD and vitamin A supplementation on immune responses to a pentavalent rotavirus vaccine, RotaTeq® in a neonatal gnotobiotic pig model. Vaccine efficacy was assessed against virulent G1P[8] human rotavirus (HRV) challenge. VAD and vitamin A sufficient (VAS) piglets were derived from dietary VAD and VAS sows, respectively. VAD piglets had significantly lower levels of hepatic vitamin A compared to that of VAS piglets. RotaTeq®-vaccinated VAD piglets had 350-fold higher fecal virus shedding titers compared to vaccinated VAS piglets post-challenge. Only 25% of vaccinated non-vitamin A supplemented VAD piglets were protected against diarrhea compared with 100% protection rate in vaccinated non-supplemented VAS piglets post-challenge. Intestinal HRV specific immune responses were compromised in VAD piglets. Vaccinated VAD piglets had significantly lower ileal HRV IgG antibody secreting cell (ASC) responses (pre-challenge) and duodenal HRV IgA ASC responses (post-challenge) compared to vaccinated VAS piglets. Also, intestinal HRV IgA antibody titers were 11-fold lower in vaccinated VAD compared to vaccinated VAS piglets post-challenge. Persistently elevated levels of IL-8, a pro-inflammatory mediator, and lower IL-10 responses (anti-inflammatory) in vaccinated VAD compared to VAS piglets suggest more severe inflammatory responses in VAD piglets post-challenge. Moreover higher IFN-γ responses pre-challenge were observed in VAD compared to VAS piglets. The impaired vaccine-specific intestinal antibody responses and decreased immunoregulatory cytokine responses coincided with reduced protective efficacy of the RV vaccine against virulent HRV challenge in VAD piglets. In conclusion, VAD impaired antibody responses to RotaTeq® and vaccine efficacy. Oral supplementation of 100,000IU vitamin A concurrent with RV vaccine failed to increase the vaccine efficacy in VAD piglets.

Interpretation of Real-time PCR Diagnosing Fecal Viruses in Children

Interpretation of Real-time PCR Diagnosing Fecal Viruses in Children

Pathogenesis of GIII.2 bovine norovirus, CV186-OH/00/US strain in gnotobiotic calves

The pathogenesis of GIII.2 bovine norovirus (BoNoV) is not well understood. Our study demonstrated persisting diarrhea and prolonged fecal shedding, but with a lack of significant intestinal lesions in gnotobiotic (Gn) calves infected with GIII.2 BoNoV, CV186-OH/00/US strain. Nine 4 to 7-day-old Angus/Jersey crossbred Gn calves were orally inoculated with 10.0–11.9log10 genomic equivalents (GE)/calf of CV186-OH (n=7) or mock (n=2). Calves were euthanized at post-inoculation day (PID) 1 (n=1) when moderate to severe lethargy was observed and at PIDs 2–6 (n=4) after lethargy had subsided. Two calves were kept longer term (until PID 30) for monitoring fecal shedding patterns by TaqMan real-time RT-PCR (qRT-PCR). Most infected calves exhibited two clinical signs: (i) acute but persisting diarrhea and (ii) acute moderate to severe lethargy. The two infected calves, followed longer-term, had prolonged fecal viral RNA shedding [peak average titer of 11.8 (±0.2)log10GE/ml] at least until PID 20. By qRT-PCR, 5 infected calves had low viral RNA titers in serum, ranging from 4.0 to 5.8log10GE/ml, at PIDs 1–5, but not (<2.7log10GE/ml) at PIDs 6–30. The latter observation coincided with the presence of serum IgG antibody to BoNoV at PIDs 8–30. Collectively, the GIII.2 BoNoV strain CV186-OH induced only mild enteropathogenicity, evident by the lack of significant intestinal lesions, but it led to persisting mild diarrhea and prolonged fecal virus shedding in Gn calves. The prolonged fecal shedding of GIII.2 BoNoV might partially explain how this virus is maintained as endemic infections in cattle.

Detection and characterization of potentially zoonotic viruses in faeces of pigs at slaughter in Germany

Pigs can harbour a variety of viruses in their gastrointestinal tract. Some of them are closely related to human viruses and are therefore suspected to have a zoonotic potential. Only little is known about the presence of those viruses in pigs at slaughter. However, by contamination of meat with zoonotic viruses during the slaughtering process, food-borne transmission to humans may be possible. Here we analyzed 120 faecal samples of pigs at slaughter from 3 different geographical regions of Germany for the presence of astrovirus (AstV), encephalomyocarditis virus (EMCV), hepatitis E virus (HEV), norovirus genogroup II (NoV GII) and group A rotavirus (GARV). Using real-time RT-PCR, the most frequently detected virus was AstV, which was present in 20.8% of the samples, followed by NoV GII with a detection rate of 14.2%. EMCV, HEV and GARV were found only occasionally with detection rates of 4.2%, 2.5% and 0.8%, respectively. Analyses of partial genome sequences of the viruses indicated that the detected AstV and NoV GII mainly represented typical pig virus strains, which have not been detected in humans so far. However, the GARV and HEV strains were more closely related to human strains. The results indicate that enteric viruses, some of them with zoonotic potential, are present in pig faeces at slaughter. Application of good hygiene practice is necessary to minimize the risk of introducing these viruses into the food and to prevent virus transmission to highly exposed persons such as slaughterers and veterinarians.

Microbiological quality of human consumption water in a school in Maringa-PR

Contaminated water associated with poor sanitation kills about 1.6 million people worldwide annually. Lack of potabilityin water is not always perceptible to the sight or smell, requiring laboratory analysis to detect it. Because of that it is importantto control microbiologically water since it is a vehicle of transmission of bacteria such as total and fecal coliforms,protozoa, viruses and fungi that cause numerous humans diseases. These microorganisms cause diarrhea, dysentery, hepatitis,cholera, among other serious illnesses. This research aimed at examining the presence of total and fecal coliformsin samples from bottled mineral water and treated water. In the present study 50 sites of drinking water were selected, alllocated in an educational institution in Maringa-PR. 13 of the collected samples were from water drinker dispensers withbottled natural mineral water and 37 samples were from water dispensers with water treated in their own institution. Themethodology used for detecting coliforms was the method of Multiple Tube Fermentation. The result of this study revealed– through microbiological analysis – that the samples of treated water were fit for human consumption. However, 15.38%of the analyzed mineral water samples were contaminated with total coliforms. These results showed that water treatedwith chlorine showed better microbiological quality when compared to bottled natural mineral water.

Effect of Zataria multiflora essential oil on immune responses and faecal virus shedding period in broilers immunized with live Newcastle disease vaccines

Summary Zataria multiflora essential oil possesses immunomodulatory effects and the present study investigates its effect on immune responses to live Newcastle disease (ND) vaccines and faecal shedding period of vaccine strain virus in chickens. One hundred and eighty chickens from both sexes were divided into 6 equal groups: group 1: unvaccinated, group 2: vaccinated (B1 vaccine followed by La Sota 10 days later), group 3: vaccinated + levamisole (150 mg/kg/day, orally), groups 4, 5 and 6 were vaccinated and received Z. multiflora essential oil by SC injections at 0.1, 0.2 and 0.4 g/kg/day dosages, respectively. Levamisole and Z. multiflora essential oil were administered for 10 and 5 consecutive days post B1 vaccination. On days 0, 7, 14, 28 and 35 post booster vaccination (PBV), HI test was performed. Delayed type hypersensitivity (DTH) test was accomplished on day 14 PBV. Faecal virus shedding was determined during 15 days PBV with 3 day intervals by RT-PCR method. Zataria multiflora essential oil induced a dose-dependent increase in antibody titers in an extent higher than levamisole as well as a dose-dependent suppression of DTH reaction. Effect of Z. multiflora essential oil at the dosage of 0.1 g/kg on shortening of faecal virus shedding period was exactly the same as levamisole where higher doses demonstrated stronger decrease. In conclusion, Z. multiflora essential oil stimulates humoral immune responses and shortens faecal virus shedding period while suppressing cell-mediated immune responses in chickens vaccinated with live ND vaccines.

Hepatobiliary Quiz-7 (2013)

1.Regarding the molecular virology of hepatitis E virus (HEV) the following are TRUE:1.HEV genome consists of three open reading frames (ORF)2.ORF1 codes for structural proteins3.The 5′ untranslated region is important for viral encapsidation4.The 3′ untranslated region is important for initiation of viral replication5.Unlike other positive strand ribonucleic acid (RNA) viruses, replication of HEV does not require a negative sense RNA intermediary2.Regarding the immunology of HEV the following are TRUE:1.First sign of infection is the detection of IgM antibodies2.Shedding of virus in feces commonly persists after onset of clinical disease3.IgG antibodies persist for years after infection and may protect from future infection4.Liver damage is immune mediated rather than due to direct cytotoxic effect of HEV5.Antibody mediated liver injury is more important than cellular mechanisms3.Regarding chronic HEV infection the following are TRUE:1.Chronic HEV infection is exclusively seen with genotype 3 in immunocompromised individuals2.HEV RNA positivity in solid organ transplant recipients is about 30%3.HEV infection in immunocompromised patients is severe with higher bilirubin and aminotransferase levels4.Chronic HEV infection can progress to cirrhosis5.Risk factors for chronic HEV infection include use of tacrolimus, higher degree of immunosuppression and large heterogeneity of quasispecies of HEV4.Regarding immunosuppressive agents used after liver transplantation the following are TRUE EXCEPT:1.Tacrolimus is more likely to cause diabetes than cyclosporine2.Gingival hyperplasia and hypertrichosis are common with tacrolimus3.Main advantage of using mycophenolate is absence of haematological side effects4.Sirolimus is has additional anti-tumour effects making it useful in cases of transplantation for hepatocellular carcinoma5.Basiliximab and daclizumab are IL-2receptor blocking antibodies5.Regarding involvement of the liver in sarcoidosis the following are TRUE EXCEPT:1.Majority of patients of sarcoidosis will have liver involvement on investigation2.Symptomatic liver disease is common in sarcoidosis3.Pulmonary involvement is absent in about 25% of hepatic sarcoidosis4.Portal hypertension is a common finding in hepatic sarcoidosis5.Sarcoidosis is not a cause for cirrhosis of the liver6.Regarding non-alcoholic fatty liver disease (NAFLD) in Asia the following are TRUE:1.South Asians like Indians are at a higher risk of insulin resistance than their Western counterparts2.India has a low prevalence of NAFLD3.Prevalence of NAFLD is increasing in Asian countries4.PNPLA3 gene polymorphisms are not associated with NAFLD in Asians5.APO C3 gene polymorphisms are associated with NAFLD predominantly in Caucasians7.Regarding liver transplantation for cirrhosis due to non-alcoholic steatohepatitis (NASH) the following are TRUE EXCEPT:1.Recurrence of NAFLD is very common2.Overall survival after transplantation is poorer compared to other etiologies of cirrhosis3.Renal dysfunction after transplantation is less common compared to other etiologies of cirrhosis4.Cardiovascular events are a more common cause of death after transplantation in NASH patients5.Pre-transplant diagnosis of NASH related cirrhosis is an independent predictor of development of metabolic syndrome after transplantation8.Regarding nutritional management of patients with cirrhosis the following are TRUE:1.Resting energy expenditure is lower in patients with cirrhosis2.Late evening snack of complex carbohydrates may improve quality of life and survival3.Protein restriction is not advisable even in patients with hepatic encephalopathy4.Animal protein is preferred to vegetable protein5.Branched chain amino acids improve event free survival9.Regarding management of ascites in cirrhosis the following are TRUE:1.Mean arterial pressure is a predictor of survival in patients of cirrhosis with ascites2.Angiotensin converting enzyme inhibitors are good anti-hypertensive drugs in patients with ascites3.Standard medical therapy consisting of sodium restriction and diuretics is successful in controlling ascites in 90% of cases4.Midodrine is a treatment option for patients with refractory ascites5.Peritoneovenous shunts are a favoured treatment option for refractory ascites10.Regarding spontaneous bacterial peritonitis (SBP) the following are TRUE:1.Culture-negative neutrocytic ascites has better outcome than culture positive SBP2.Monomicrobial non-neutrocytic bacterascites frequently resolves spontaneously3.Albumin infusions decrease mortality in patients with SBP4.Primary prophylaxis for SBP is indicated if Child score is ≥9 and serum bilirubin is ≥3 mg/dl5.Short-term prophylaxis for SBP in patients with gastrointestinal bleeding reduces infection rate but does not affect mortality

Infectivity of a genotype 4 hepatitis E virus cDNA clone by intrahepatic inoculation of laboratory rats

A cDNA clone of a genotype 4 swine hepatitis E virus (HEV) strain (SAAS-FX17), identified in Shanghai, has been constructed. Capped RNA transcripts were prepared in vitro and shown to be replication-competent in Huh7 cells. Sprague-Dawley (SD) rats administered the RNA transcripts by intrahepatic inoculation developed active infections as evidenced by fecal virus shedding and sero-conversion to anti-HEV. The former was first detected between 23 and 30 days post-inoculation (dpi) and persisted until 45 dpi. Sera of rats inoculated with RNA transcripts became anti-HEV positive between 30 and 40 dpi, and reverted to anti-HEV negative at 52 dpi. Our data indicate for the first time that intrahepatic inoculation of rats with RNA transcripts of an HEV cDNA clone may serve as an alternative animal model for HEV research.

Detection of bovine corona virus in some governorates of Iraq

In the present study, we used Reverse transcription polymer chain reaction (RT-PCR) assay for detecting of bovine coronavirus, We evaluated the presence of bovine coronavirus ( BCoV) in diarrheic fecal samples in age (1 – 30) day. (152) fecal samples from diarrheic calves were collected from four Iraqi governorates (Al-Qadisyiah, Babylon, Wassit, and Najaf) .10 (6.57%) out of 152 were positive to bovine coronavirus. This study is the first detection of bovine corona antigen in Iraq.T he results suggest that RT-PCR is more sensitive than another method to detect BCoV, especially in subclinical cases and Because these animals shed a low amount of virus in feces and more animal show clinically normal we need to apply sensitive techniques, such as RT-PCR.

Rabbit and human hepatitis E virus strains belong to a single serotype

Hepatitis E virus (HEV) is a zoonotic pathogen and all four established genotypes of HEV belong to a single serotype. The recently identified rabbit HEV is antigenically and genetically related to human HEV. It is unclear whether rabbit HEV belongs to the same serotype as human HEV. The purpose of this study was to determine the serotypic relationship between rabbit and human HEVs. HEV ORF2 recombinant capsid protein p166 (amino acids 452–617) of four known HEV genotypes and rabbit HEV were used to induce immune serum, which were evaluated for their ability to neutralize human HEV genotype 1, 4, and rabbit HEV strains by an in vitro PCR-based HEV neutralization assay. Immune sera of five kinds of p166 proteins were all found to neutralize or cross-neutralize the three different HEV strains, suggesting a common neutralization epitope(s) existing between human and rabbit HEV. Rabbit models of a second-passage rabbit HEV strain, JS204-2, and a genotype 4 human HEV strain, NJ703, were established as evidenced by fecal virus shedding, viremia and anti-HEV IgG seroconversion. Six rabbits, recovered from JS204 infection, were challenged with NJ703, and another six recovered from NJ703 infection were challenged with JS204-2. After challenge, viremia was not detected, shorter fecal virus shedding durations and obvious early stage declines in anti-HEV IgG values were observed. The results from this study indicate that rabbit HEV belongs to the same serotype as human HEV.

Survivability of Highly Pathogenic Avian Influenza H5N1 Virus in Poultry Faeces at Different Temperatures

Highly pathogenic avian Influenza (HPAI) is an important zoonotic disease and is becoming a great threat to poultry industry. India has experienced continual outbreaks of H5N1 HPAI virus since February, 2006 especially in Eastern India. Survivability in poultry faeces is an important determinant in evaluating the persistence of the virus in the poultry sheds and their vicinity. In this paper, survivability of Indian H5N1 HPAI virus in dry and wet poultry faeces at 42, 37, 24 and 4°C, respectively is reported. The effect of different temperatures was determined by linear regression model and defined in terms of linear equation. The virus survived up to 18h at 42°C, 24h at 37°C, 5days at 24°C and 8weeks at 4°C in dry and wet faeces, respectively. The coefficients of determination (R(2)) values for dry and wet faeces revealed that the difference in viral persistence in dry and wet faeces at all temperatures was not very marked. Results of the present study indicated that H5N1 HPAI virus may remain viable for extended periods of time in faeces at low temperatures and may act as a long term source of influenza virus in the environment.

Evaluation of the Bioline Standard Diagnostics SD immunochromatographic norovirus detection kit using fecal specimens from Australian gastroenteritis incidents

Human norovirus is a major cause of both sporadic cases and outbreaks of gastroenteritis and comprises two main genogroups (GI and GII) which, in turn, comprise a variety of genotypes. The current study examined the efficacy of the Bioline SD kit using fecal material from Australian gastroenteritis incidents. At best, the SD kit had a sensitivity of 62%. Freezing and thawing specimens before testing significantly improved sensitivity. The SD kit had a specificity of 98.6%. Genotype analysis (Open Reading Frame 2) indicated the SD kit could detect a range of genotypes and genotype variants including GI.1, GI.3, GI.4, GII.1, GII.3, GII.4 (unclassified), GII.4 (2006b), GII.4 (2009), GII.4 (2012) and GII.6 but the kit failed to detect GI.2 and GII.2 norovirus. The kit did not cross-react with a number of common fecal viruses including astrovirus, sapovirus, rotavirus or adenovirus. The kit was very easy to use and would be valuable in point-of-care testing.