Incidence of hepatitis A and hepatitis E viruses and norovirus and rotavirus in fish and shrimp samples caught from the Persian Gulf / [Incidência de vírus da hepatite A e hepatite E e norovírus e rotavírus em amostras de peixes e camarões capturados no Golfo Pérsico]

Foodborne viruses including hepatitis A virus (HAV), norovirus (NoV), rotavirus (RoV) and hepatitis E virus (HEV) are easily transmitted through contaminated seafoods. The current research was done to assess the incidence of RoV, NoV GI and GII,hAV and hEV in fish and shrimp samples caught from the Persian Gulf, Iran. Three-hundred and twenty fish and shrimp samples were collected. The presence of foodborne viruses were assessed by the real-time PCR. Forty-nine out of 320 (15.31%) fish and shrimp samples were positive for foodborne viruses. Distribution of hAV, NoV GI and NoV GII amongst all studied samples were 0.93%, 5.93% and 8.43%, respectively. hEV and RoV viruses were not found in studied samples. Parastromateus niger and Scomberomorus commerson fish and Penaeus monodon shrimp were the most frequently contaminated samples. Simultaneous incidence of hAV and NoV GI and hAV and NoV GII were 0.31% and 0.93%, respectively. Distribution of foodborne viruses in samples collected through spring, summer, autumn and winter seasons were 14.28%, 9.33%, 11.76% and 24.44%, respectively. Findings revealed that the incidence of foodborne viruses was significantly associated with seafood species and also season of sampling.(AU)

Vírus transmitidos por alimentos, incluindo hepatite A (HAV), norovírus (NoV), rotavírus (RoV) e hepatite E (HEV) são facilmente transmitidos através de frutos do mar contaminados. Esta pesquisa foi realizada para avaliar a incidência de RoV, NoV GI e GII, hAV e hEV em amostras de peixes e camarões capturadas no Golfo Pérsico, Irã. Foram coletadas 300 amostras de peixes e camarões. A presença de vírus transmitidos por alimentos foi avaliada por PCR em tempo real. Quarenta e nove das 320 amostras de peixes e camarões (15,31%) foram positivas para vírus transmitidos por alimentos. A distribuição de hAV, NoV GI e NoV GII entre as amostras estudadas foi 0,93%, 5,93% e 8,43%, respectivamente. Os vírus hEV e RoV não foram encontrados nas amostras estudadas. Os peixes Parastromateus niger e Scomberomorus commerson e o camarão Penaeus monodon foram as amostras mais frequentemente contaminadas. A incidência simultânea de hAV e NoV GI, e hAV e NoV GII foi de 0,31% e 0,93%, respectivamente. A distribuição dos vírus transmitidos por alimentos nas amostras coletadas na primavera, verão, outono e inverno foi de 14,28%, 9,33%, 11,76% e 24,44%, respectivamente. Os resultados demonstram que a incidência de vírus transmitidos por alimentos foi significativamente associada às espécies de frutos do mar e também à época da amostragem.(AU)

SEROLOGICAL DETECTION OF HEPATITIS A VIRUS IN FREE-RANGING NEOTROPICAL PRIMATES (Sapajus spp., Alouatta caraya) FROM THE PARANÁ RIVER BASIN, BRAZIL

Nonhuman primates are considered as the natural hosts of Hepatitis A virus (HAV), as well as other pathogens, and can serve as natural sentinels to investigate epizootics and endemic diseases that are of public health importance. During this study, blood samples were collected from 112 Neotropical primates (NTPs) (Sapajus nigritus and S. cay, n = 75; Alouatta caraya, n = 37) trap-captured at the Paraná River basin, Brazil, located between the States of Paraná and Mato Grosso do Sul. Anti-HAV IgG antibodies were detected in 4.5% (5/112) of NTPs, specifically in 6.7% (5/75) of Sapajus spp. and 0% (0/37) of A. caraya. In addition, all samples were negative for the presence of IgM anti-HAV antibodies. These results suggest that free-ranging NTPs were exposed to HAV within the geographical regions evaluated.

The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge

The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

Hepatitis C and b virus in schistosomiasis patients on oral or parenteral treatment

Egypt leads the world in hepatitis C infection cases. Schistosomiasis and hepatitis C virus [HCV] co-infection is common in Egypt. Hepatitis B [HBsAg] is also encountered. Potassium antimony tartarate was used for Schistosomiasis in endemic areas all over the world for more than 50 years, intravenously and was prohibited since late of the last century when the oral drugs were developed. Some authors postulated that Schistosomiasis might contribute to persistence of HCV via biological factors. This study compared the prevalence of HCV and HBV among Schistosomiasis patients in endemic areas given parenteral or oral route anti-schistosomiasis. A total of 102 manual workers patients were selected from different areas in Cairo and Gharbia Governorates. They were 82 males and 20 females; their age was ranged between 16-60 years [40 +/- 12]. Sheets were filled out on each patient including name, age, sex, history of haematurea or bloody diarrhea, parasitic infection, intravenous injections, blood transfusion, dental care, shaving, previous treatment, and anti-schistosomiasis. They were divided into GI: 50 patients treated IV since 20-30 years ago with ages [49.4 +/- 7.4] and GIL 52 patients treated orally up to 8 years ago with ages [2.2 +/- 7.7]. They were subjected to IHAT anti-schistosomal antibodies, circulating schistosome antigens in urine, HBsAg and HCVAb, the latter was confirmed by PCR for positivity, and Alanin transeferese [ALT]. In GI, 42/50 [84%] had HCV compared to 4/52[7.7%] in Gil, with statistical significant. HBsAg was detected in a patient. HCV patients showed function. 3/52 received oral treatment showed elevation of liver function [5.8%]. 10% [10/50] of GI and 34.6% [18/52] of Gil were schistosomal antigen in urine positive

Investigation of a hepatitis A outbreak from Shimla Himachal Pradesh.

Background & objective: Hepatitis A is an enterically transmitted viral disease, highly prevalent in India and mainly presents as a paediatric sporadic disease. This study investigated an outbreak of viral hepatitis at Shimla, Himachal Pradesh, India, during January-March 2007. Methods: Eighty seven blood samples, 3 water samples and 2 sewage samples were collected. Serum samples were tested for IgM and IgG anti HAV and IgM and IgG anti HEV antibodies. Serum, sewage and water samples were tested for HAV-RNA by nested RT- PCR. Nearly complete full genome (excluding extreme 5’ end) was amplified from one serum sample. Results: The hepatitis cases were mainly seen among children and young adults and 63.2 per cent (55/88) were positive for anti-HAV IgM. These cases were reported from the areas getting water supply from Ashwani Khud water supply system. This water purification system received water from a natural stream in which treated sewage water was let into 4 km upstream the collection point since one year. HAV-RNA present in serum, sewage and water samples showed 100 per cent sequence homology. Phylogenetic analysis based on 5’ non coding (5’ NC) and nearly complete genome showed the evidence of HAV genotype IIIA in all the samples. Interpretation and conclusion: The aetiological agent of the present outbreak was hepatitis A virus which is emerging in an outbreak form in India, emphasizing a definite need for formulating vaccination / control strategies.

Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods

The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20 percent of the AdV, 90 percent of the RV and 100 percent of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8 percent of the AdV and 90 percent of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.

Molecular characterization of hepatitis A virus isolates from Goiânia, Gioás, Brazil

Hepatitis A virus (HAV) infection is a public health problem worldwide and the virus has been classified into six genotypes. In Brazil, the only genotype that has been found is genotype I, predominately from subgenotype IA. Here, the HAV genotypes were analyzed of 18 isolates circulating between 1996-2001 in Goiânia, state of Goiás, Brazil. Viral RNA was extracted from 18 serum samples and amplified (RT-PCR/nested-PCR), followed by the genomic sequencing of the VP1/2A junction region of the HAV genome. Sequences of 168 nucleotides were compared and analyzed using the BLAST N, Clustal X and PAUP v. 4.10b programs. All samples were classified as genotype I, with 10 belonging to subgenotype IA and eight to subgenotype IB. The subgenotype IA isolates showed greater diversity than the subgenotype IB isolates at the nucleotide level. Elevated identity values were found between isolates obtained in this study and those from other regions of the world, including Brazil, highlighting the high conservation among different isolates of this virus. However, changes in the HAV subgenotype circulation could also be observed during the evaluated period.

Estimativa de áreas de risco para hepatite A / Estimation of risk areas for hepatitis A

Este estudo estimou áreas de risco de hepatite A e de variáveis associadas em região de Duque de Caxias, Rio de Janeiro, Brasil. Foi realizado um inquérito soroepidemiológico para hepatite A e domiciliar em 19 setores censitários. Destes, foram selecionados 11 setores contíguos com diferentes cotas altimétricas e todas as 1.298 crianças com idade menor que dez anos foram incluídas no estudo. Construiu-se semivariogramas para a hepatite A, não uso de filtro, número de pontos de água e tempo de encanamento de água. A estimativa espacial foi obtida por krigagem indicadora e ordinária. O ajuste dos modelos foi avaliado por meio de validação cruzada. Os quatro modelos ajustados de semivariogramas do desfecho e das três variáveis sócio-econômicas apresentaram um padrão isotrópico; mapas do risco estimado segundo a krigagem para hepatite A e para as variáveis sócio-econômicas foram construídos; a validação cruzada mostrou um bom ajuste. A utilização do método de estimativa espacial usando-se a krigagem possibilitou a detecção de áreas com maior probabilidade de ocorrência da hepatite A, independentemente da divisão político-administrativa dos setores censitários.

This study estimated hepatitis A risk areas in a region of Duque de Caxias, Rio de Janeiro State, Brazil. A cross-sectional study consisting of a hepatitis A serological survey and a household survey were conducted in 19 census tracts. Of these, 11 tracts were selected and 1,298 children from one to ten years of age were included in the study. Geostatistical techniques allowed modeling the spatial continuity of hepatitis A, non-use of filtered drinking water, time since installation of running water, and number of water taps per household and their spatial estimation through ordinary and indicator kriging. Adjusted models for the outcome and socioeconomic variables were isotropic; risk maps were constructed; cross-validation of the four models was satisfactory. Spatial estimation using the kriging method detected areas with increased risk of hepatitis A, independently of the urban administrative area in which the census tracts were located.

Co-circulation of genotypes IA and IB of hepatitis A virus in Northeast Brazil

The Northeast region is the location of most cases of acute hepatitis A virus (HAV) in Brazil. In the present study, the genotypes of HAV strains from Pernambuco State, one of most populous states in the Northeast region, were characterized. Blood samples positive for anti-HAV IgM from 145 individuals (mean age = 29.1 years), collected during 2002 and 2003, were submitted to nested RT-PCR for amplification of the 5'non-translated region (5'NTR) and VP1/2A regions of the HAV genome. The VP1/2A and 5'NTR regions were amplified in 39 and 21 percent of the samples, respectively. Nucleotide sequencing was carried out in 46 percent of VP1/2A and in 53 percent of 5'NTR isolates. The identity in nucleotide sequence of the VP1/2A region ranged from 93.6 to 100.0 percent. Phylogenetic analysis of the VP1/2A sequences showed that 65 percent belong to sub-genotype IA and 35 percent to sub-genotype IB. Co-circulation of both sub-genotypes was observed in the two years studied. Distinct clusters of highly related sequences were observed in both sub-genotypes, suggesting endemic circulation of HAV strains in this area. In the 5'NTR isolates, 92.7-99.2 percent identity was observed and two isolates presented one deletion at position 413. Phylogenetic analysis showed that genotype IA strains cluster in the tree in the same way as genotype IB strains, but one IIIA isolate from Spain clusters with genotype IB strains. These results do not allow us to state that 5'NTR could be used to genotype HAV sequences. This is the first report of co-circulation of sub-genotypes IA and IB in this region, providing additional information about the molecular epidemiology of HAV strains in Brazil.

Actualidad en hepatitis por virus A / Current situation in viral hepatitis A

La hepatitis por virus A (VHA) es una de las enfermedades más ampliamente difundidas en el mundo, generalmente aparece en forma de brotes epidémicos y se trasmite predominantemente por vía fecal oral, un tercio de los casos reportados por esta infección ocurre en niños y consta de varias formas clínicas de presentación; el tratamiento se basa en la aplicación de medidas generales así como la inmunoprofilaxis activa y pasiva, su elevada prevalencia en nuestro medio nos motiva a revisar la literatura reportada hasta hoy sobre el tema y resumir los aspectos más novedosos en cuanto a sus características y formas de evitarla.

Comparison of four extraction methods to detect hepatitis A virus RNA in serum and stool samples

The efficiency of extraction methods for hepatitis A virus (HAV) RNA in clinical samples is of great importance for molecular diagnosis, especially in regions endemic for HAV, such as Brazil. We compared the efficiency of four different extraction techniques in serum and stool samples for the detection of hepatitis A virus by reverse transcription PCR (RT-PCR). We used PCR to analyse serum and stool samples of 12 patients who were referred to the Brazilian Reference Center for Viral Hepatitis (BRCVH) in Rio de Janeiro. The methods tested were Proteinase K, Silica, TRIzol and Guanidine isothiocyanate. Proteinase K extraction was the best method for serum samples; it detected the HAV-RNA in 11 of the 12 samples. The guanidine isothiocyanate method was the most effective for stool samples, detecting HAV-RNAs in 9 of the samples. The TRIzol« method worked best with serum samples, and the silica method was unsatisfactory with both serum and stool samples. The RNA extraction method affected the outcome. The use of appropriate RNA extraction methods is a critical step for successful and valid PCR studies on clinical samples. We recommend that RNA extraction techniques be carefully selected for their efficiency with each type of specimen