ABSTRACT New viruses of the Picornavirales order have been discovered with the increase in the number of sequences obtained by high-throughput sequencing, as well as human stool-associated RNA virus (husavirus [HuV]), found in human stool samples. However, there is much to be clarified about HuV. Its cellular host, evolutionary history, and other biological characteristics are still unknown. Therefore, samples collected from human beings and environmental samples in a watershed in Southern Brazil were processed for the metagenomic library. Upon metagenomic analysis, we identified a HuV (husavirus LMM_67754 OP019707) genome with 8,846 bp, which was reported for the first time in Southern Brazil. The new genome presents only 37% of nucleotide identity with Brazilian strains and more than 90% with genomes from China, Vietnam, Venezuela, and the Netherlands. The HuV phylogeny presents significant differences among genomes, probably because multiple introductions of the virus may have occurred. Many questions still need to be answered about HuV. Therefore, more sequences and studies on this virus are necessary to improve the comprehension of the unknown origin of Picornavirales.

Abstract December 2019 marked the beginning of the current Coronavirus disease pandemic (COVID-19). Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) was identified as the causative agent of a viral pneumonia outbreak in Wuhan, Hubei Province, China. The alarming spread levels and clinical severity elevated the status of COVID-19 to the global pandemic by the World Health Organization. In 6 months, more than 25 million cases of infected people and more than 890,000 deaths by COVID-19 had been reported worldwide. The main goal of this review is to shed light upon the current COVID-19 epidemic situation in Brazil with a health approach highlighting some unique environmental, animal and epidemiological aspects.

ABSTRACT Multiple variants of the Severe Acute Respiratory Syndrome coronavirus 2 virus (SARS-CoV-2) have been constantly reported across the world. The B.1.1.28 lineage has been evolving in Brazil since February 2020 and originated the P.1 variant of concern (VOC), recently named as the Gamma variant by the newly WHO nomenclature proposal, and P.2 as a variant of interest (VOI). Here we describe an early case of P.1 primary infection in Southern Brazil in late November 2020, soon after the emergence of the variant in Manaus, Northern Brazil. The same male patient was reinfected by another B.1.1.28 variant, namely P.2, in March, 2021. The genomic analysis confirmed genetically significant differences between the two viruses recovered in both infections, the P.1 lineage in the first episode and P.2 in the reinfection. Due the very early detection of P.1, we have also investigated the circulation of P.1 in the same region by differential RT-qPCR, showing that this was an isolated case of P.1 at the time of detection, and this variant has disseminated and became prominent from late January to the end of March, 2021. SARS-CoV-2 recent reports of reinfection have raised critical questions on whether and how well a first infection protects against reinfection.

RESUMO: As doenças transmitidas por alimentos são frequentemente descritas e relacionadas ao consumo de alimentos ou água contaminados, sendo alguns agentes virais importantes fontes de contaminação e frequentemente encontrados em alimentos de origem animal. O objetivo deste trabalho foi detectar patógenos entéricos emergentes em amostras de alimentos industrializados de origem animal coletados em estabelecimentos da região sul do Brasil. Nas amostras analisadas, não foi detectado o genoma do vírus da Hepatite E (HEV). No entanto, 21,8% (21/96) das amostras foram positivas para Rotavírus (RV) e 61,4% (59/96) para adenovírus (AdV), incluindo Adenovírus humano-C (HAdV-C), Adenovírus porcino-3 (PAdV-3) e novo tipo de suíno adenovírus PAdV-SVN1. No presente trabalho, é descrito pela primeira vez em alimentos a presença de PAdV-SVN1. A presença desses vírus pode estar relacionada à falta de higiene em locais de preparo de alimentos, manipulação de produção.

ABSTRACT: Foodborne diseases are often related to consumption of contaminated food or water. Viral agents are important sources of contamination and frequently reported in food of animal origin. The goal of this study was to detect emerging enteric viruses in samples of industrialized foods of animal origin collected in establishments from southern of Brazil. In the analyzed samples, no Hepatitis E virus (HEV) genome was detected. However, 21.8% (21/96) of the samples were positive for Rotavirus (RVA) and 61.4% (59/96) for Adenovirus (AdV), including Human adenovirus-C (HAdV-C), Porcine adenovirus-3 (PAdV-3) and new type of porcine adenovirus PAdV-SVN1. In the present research, PAdV-SVN1 was detected in foods for the first time. The presence of these viruses may be related to poor hygiene in sites of food preparation, production or during handling.

ABSTRACT Public parks are leisure environments widely used by both, adults and children, often accompained by their pets. Soil contamination of these environments by enteric viruses and intestinal parasites occurs through these animals feces. The aim of this work was to detect Carnivore protoparvovirus 1 (CPV-1) and different species of Mastadenovirus in soils samples from a park located in a medium-sized city in Brazil and evaluate the presence of helminth eggs and larvae in 18 points of a public park soil samples, as well as feces found on this site during six months. Parasitological analyzes were conducted through flotation and sedimentation techniques, and polymerase chain reaction (PCR) was used for viral detection. Of the 216 soil and 16 feces samples, 49% (106/216) and 12% (2/16) were positivefor nematodes larvae, respectively, through sedimentation techniques. Toxocara spp eggs were found in one soil sample and one feces sample, Trichuris spp eggs were found in only one feces sample and Hookworms eggs were found in four soil samples. After reconstruction work in the streets near the park, 30% (64/216) of the samples were positive for Human Mastadenovirus C (HAdV-C), 1.4% (3/216) for HAdV-E and 0.4% (1/216) for Canine Mastadenovirus A (CAdV-A). The parasitic forms found in this study have demonstrated that the contamination of the park’s soil pose a threat to human and animal health. This was the first study to report the presence of HAdVs and CAdVs in soil samples.

Resumo A destilação natural da água pode destruir e/ou inativar microorganismos que são sensíveis ao calor e a radiação ultravioleta (UV). Este método é utilizado, atualmente, para fornecer água potável em navios e para dessalinização de água salobra. Para o desenvolvimento desta pesquisa, um destilador solar em escala piloto foi construído e instalado na região sul do Brasil de forma a se verificar a eficiência do equipamento na desinfecção de água. A verificação de eficiência do equipamento foi baseada na determinação do número mais provável (NMP) de coliformes totais e Escherichia coli bem como no número de cópias de DNA do adenovírus humano tipo 5 (HAdV-5) em amostras de água antes e após a destilação. Os resultados demonstraram que houve 100% de remoção de coliformes totais e E. coli e 4,5 log (99,997%) de remoção de HAdV-5 após a destilação, o que está de acordo com o padrão microbiológico de potabilidade determinado pela legislação brasileira, pela USEPA e pela HEALTH CANADA.

Abstract Natural water distillation can destroy and/or inactivate microorganisms that are sensitive to heat and ultraviolet radiation (UV). This method is currently used to provide fresh water in ships and in the desalination of brackish water. For the development of this research, a pilot-scale solar still was built and installed in the southern region of Brazil, in order to assess its efficiency in water disinfection, which was based on the most probable number (MPN) of total coliforms and Escherichia coli, in addition to the DNA copy number of human adenovirus type 5 (HAdV-5) in raw, undistilled samples and in treated distilled water. Results showed that the distillation process removed 100% of total coliform and Escherichia coli and 4.5 log (99.997%) of HAdV-5, which meets the microbiological standards for drinking water according to national Brazilian regulations, as well as USEPA and HEALTH CANADA requirements.

ABSTRACT Hepatitis A virus (HAV), a member of Picornaviridae family, is the main causative agent of acute viral hepatitis in the world, mainly in developing countries. HAV may be present in contaminated water and food and its presence is often associated to a lesser extent with socioeconomic factors and environmental quality. The main goals in the present study were to standardize a cell culture combined to a polymerase chain reaction protocol for the detection and quantification of viral viability and analyze whether the virus could be found in water samples collected in four urban streams of Sinos River watershed. Virus recovery was assayed from known virus concentrations measured in experimentally contaminated raw and ultrapure water (MilliQ®). Recovery rates ranged from 270% in raw water to 15,000% in ultrapure water. In a second step, a qPCR coupled to a previous passage in cells, demonstrated more analytical sensitivity when compared to samples assayed without a previous passage in cell cultures. HAV genome was detected in only 1 of 84 samples analyzed, pointing to a very low occurrence of HAV in water samples in the studied region. These findings are remarkable, since no more than 5% of the domestic sewage in this area is treated pointing to a low occurrence of HAV in the population living nearby during the study period.

Resumo O abastecimento de água do município de Ivoti provém unicamente de origem subterrânea. A potabilidade da água subterrânea do município foi avaliada através da análise de marcadores bacterianos (Escherichia coli e coliformes totais) e virais (adenovírus, enterovírus e rotavírus) de contaminação. Vinte e sete amostras de água provenientes de poços artesianos, poços cavados e fontes foram coletadas entre fevereiro e abril de 2011. A determinação do número mais provável (NMP) de E. coli e coliformes totais em 100 mL foi realizada pelo método dos tubos múltiplos. Os marcadores virais foram analisados em alíquotas de 500 mL por reação em cadeia da polimerase (PCR). Coliformes totais foram encontrados em 66,6% das amostras. A contaminação encontrada para coliformes termotolerantes foi de 25,9%, 40,7% para adenovírus e 22,2% para rotavírus. Nenhuma amostra apresentou enterovírus. A presença de coliformes e de vírus nas amostras indica contaminação fecal da água subterrânea no local de estudo, situado na porção aflorante do Aquífero Guarani e alerta para a necessidade de precaução quanto ao uso destes mananciais para abastecimento público, captação individual e cultivo de hortaliças.

Abstract The water supply of the municipality of Ivoti comes solely from an underground source. The potability of groundwater in the municipality was evaluated by analyzing bacterial (Escherichia coli and total coliforms) and viral (adenovirus, enterovirus and rotavirus) markers for contamination. Twenty-seven water samples from artesian wells, dug wells and springs were collected between February and April 2011. The determination of the most probable number (MPN) of E. coli and total coliforms in 100 mL was performed by the multiple-tube method. Viral markers were analyzed in 500 mL aliquots by polymerase chain reaction (PCR). Total coliforms were found in 66.6% of samples. Contamination found for E. coli was 25.9%, 40.7% for adenovirus and 22.2% for rotavirus. Enterovirus was not found. The presence of coliform bacteria and viruses in the samples indicates fecal contamination of groundwater at the study site, located in the recharge portion of the Guarani Aquifer and suggests the need of caution regarding the use of these water sources for public water supply, individual water catchment and farming.

Resumo A gestão de bacias hidrográficas por meio de comitês é essencial na mediação entre a oferta e a demanda de água para consumo doméstico, industrial, agrícola, lazer, entre outros usos da água. Uma gestão exitosa pode ocorrer por meio da identificação e compreensão das características socioeconômicas, demográficas, entre outras, e assim formatar as práticas adotadas pelos comitês. Neste contexto, o escopo central deste estudo consistiu em analisar as variáveis intervenientes relacionadas aos aspectos geográficos, demográficos, socioeconômicos e a existência de comitê de bacias. A metodologia consiste na abordagem quantitativa, empregando-se uma pesquisa documental secundária e sua análise ocorreu por meio da aplicação da estatística descritiva e do teste de correlação de Pearson. Os principais resultados evidenciam que a explicação da existência de comitês de bacias hidrográficas relaciona-se com o tamanho da população que promove maior pressão à medida que aumenta. Além disso, evidenciou-se que a existência de comitês de bacias correlaciona-se com: crescimento econômico, quantidade de conflitos pelos múltiplos usos, coleta de esgotos e nível de degradação da qualidade ambiental. Conclui-se, portanto, que de fato existem variáveis intervenientes para a existência de comitês de bacias e que eles são necessários para a gestão eficiente destas variáveis.

Abstract Watershed management through committees is essential in mediating between water supply and demand for domestic consumption, as well as industrial, agricultural, leisure, and other uses of water. Successful management can occur through the identification and comprehension of socioeconomic and demographic characteristics, among others, and therefore, these help shape the practices adopted by the committees. In this context, the central objective of this study was to analyse the intervening variables related to the geographic, demographic, socioeconomic aspects and the existence of a basin committee. The methodology consisted of a quantitative approach, using previously published documents and analysis applying descriptive statistics and the Pearson correlation test. The main results showed that the existence of a watershed committee is correlated with the size of the population, which promotes greater pressure as it increases. In addition, it was observed that the creation of committees is related to economic growth, number of conflicts for multiple uses, sewage collection and level of environmental quality degradation. We conclude, therefore, that in fact there are intervening variables for the existence of river basin committees and that the committees are necessary for the efficient management of those variables.

Human adenoviruses (HAdV), members of the Adenoviridae family, are excreted through the fecal route and may be present in the feces of humans consuming contaminated food or water. The presence of HAdV from different serotypes in the feces of healthy individuals was already reported using conventional polymerase chain reaction; however, real-time PCR (qPCR) may reveal not only the rates of detection as well as demonstrate the viral loads excreted by healthy persons. Aiming to identify and characterize the presence of adenoviruses in stool samples, 147 fecal samples from patients with no records of diarrhea were analyzed (74 from winter season and 73 from summer) by Real-Time PCR (qPCR) assay and conventional PCR. HAdV genome was present in 43.8% (32/73) of stools samples collected during summer season and 21.6% (16/74) during winter. The rate of detection of genomic copies (gc) ranged from 4.04×102 to 6.72×105gc/g of feces among the 147 samples analyzed, of which the ranged of genomic copies of DNA HAdV was major in summer. All samples were negative when tested for rotaviruses (RV) and noroviruses (NoV) by PCR conventional and qPCR respectively. HAdV is excreted constantly by infected individuals in the absence of clinical signs and the occurrence may vary seasonally.

RESUMO Os adenovírus humanos (HAdV) são notavelmente resistentes ao ambiente. Estes agentes podem servir como indicadores efetivos de contaminação fecal, tanto quanto podem atuar como agentes causadores de diferentes doenças em seres humanos. A reação em cadeia da polimerase (PCR) e mais recentemente a PCR quantitativa (qPCR) são amplamente usadas para detecção de agentes virais em matrizes ambientais. No presente estudo, PCR e SYBR(r)Green qPCR foram comparadas para a detecção de HAdV em amostras de água (55) e sedimento (20) provenientes de nascentes, poços, açudes e arroios coletadas em propriedades leiteiras. A metodologia quantitativa detectou HAdV em 87,3% das amostras de água e 80% dos sedimentos, enquanto por PCR convencional a detecção foi de 47,3% e 35%, respectivamente.

SUMMARY Human Adenoviruses (HAdV) are notably resistant in the environment. These agents may serve as effective indicators of fecal contamination, and may act as causative agents of a number of different diseases in human beings. Conventional polymerase chain reaction (PCR) and, more recently, quantitative PCR (qPCR) are widely used for detection of viral agents in environmental matrices. In the present study PCR and SYBR(r)Green qPCR assays were compared for detection of HAdV in water (55) and sediments (20) samples of spring and artesian wells, ponds and streams, collected from dairy farms. By the quantitative methodology HAdV were detected in 87.3% of the water samples and 80% of the sediments, while by the conventional PCR 47.3% and 35% were detected in water samples and sediments, respectively.

Um novo teste baseado na reação em cadeia da polimerase em tempo real (qPCR) com SYBR ® Green foi desenvolvido para detectar duas espécies de Bartonella, B. henselae e B. clarridgeiae, diretamente em amostras de sangue. Este teste foi utilizado em amostras de sangue obtidas de gatos que vivem em abrigos de animais do sul do Brasil. Os resultados foram comparados aos obtidos pelo PCR convencional utilizado para a detecção de Bartonella spp. Das 47 amostras analisadas, oito foram positivas no PCR convencional e 12 foram positivas para qPCR. A reação de qPCR, permitiu a detecção da presença simultânea de B. henselae e B. clarridgeiae em duas destas amostras. Os resultados mostram que a qPCR aqui descrita pode ser uma ferramenta confiável para a detecção e diferenciação de duas espécies importantes de Bartonella spp.

A novel SYBR® green-real time polymerase chain reaction (qPCR) was developed to detect two Bartonellaspecies, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.

Chronic viral hepatitis are main public health problems worldwide. Data about the seroprevalence to Hepatitis B and C viruses (HBV e HCV) at the population level are scarce on Brazil and especially for the Southern region of the country. The seroprevalence to HBV and HBC antigens was evaluated on a large portion of the population of the municipality of Caxias do Sul (427,858 inhabitants), Brazil. A total of 60,604 individual serum samples collected from 2008 to 2011 were screened for HBV surface antigen (HBsAg) and for antibodies against HBsAg; anti HCV antibodies were measured by ELISA (Enzyme Linked Immunosorbent Assay). Overall, 1.63% of the individuals were positive for HBsAg and 1.43% showed seropositivity to HCV. From the total, 31,749 samples were analyzed for HBsAg and 28,855 for HCV. For HBsAg, 519 samples showed positive (1.63%) while the results for 37 patients (0.12%) remained inconclusive. For the anti-HCV test, 412 individuals (1.43%) showed positive. From the positive samples for HBsAg 216 (50.6%) were from male individuals whereas for anti-HCV the seroprevalence was slightly higher for females 216 (52.4%). The higher prevalence for both hepatitis viruses were found among individuals at the age group of 40 to 59 years and the lower levels of positivity for both HBV and HBC were among children and teenagers.

O número de vírus animais cresce continuamente, causando preocupação permanente a virologistas e veterinários. O potencial patogênico e associação com doença tem sido claramente demonstrado para alguns - mas não para todos - vírus emergentes. Esse artigo apresenta uma breve revisão das recentes descobertas de vírus animais e a sua potencial relevância para saúde animal. Cães eram considerados refratários aos vírus da influenza até 2004, quando um vírus influenza A subtipo H3N8 foi transmitido de equinos e causou doença respiratória severa em cães galgos na Flórida/EUA. O novo vírus, denominado vírus da influenza canina (CIV), agora considerado uma linhagem distinta do vírus da influenza equina, disseminou-se na população canina urbana dos EUA. Um novo Pestivirus (Flaviviridae) - provisoriamente denominado pestivírus Hobi-like - foi identificado em 2004 em soro fetal bovino importado do Brasil. Os vírus Hobi-like são genética e antigenicamente relacionados com o vírus da diarreia viral bovina (BVDV) e induzem manifestações clínicas semelhantes. A sua origem e distribuição são desconhecidas, mas estão aparentemente disseminados no rebanho brasileiro e já foram identificados no sudeste asiático e na Europa. Em 2011, um novo buniavírus transmitido por mosquitos, denominado vírus Schmallemberg (SBV), foi associado com febre, redução da produção de leite, abortos e malformações fetais em bovinos e ovinos da Alemanha. Subsequentemente, esse agente se disseminou por vários países europeus e atualmente representa uma séria ameaça para saúde animal naquele continente. A origem do SBV é um tema ainda controverso, mas possivelmente tenha resultado de ressortimento entre os buniavírus Shamonda e Satuperi. O vírus da hepatite E (HEV, família Hepeviridae) é um conhecido agente de hepatite aguda em humanos e, em 1997, foi identificado pela primeira vez em suínos. Estudos epidemiológicos posteriores revelaram que o HEV está amplamente distribuído em rebanhos suínos, principalmente associado com infecção subclínica. Dois dos quatro sorotipos do HEV são zoonóticos e podem ser transmitidos entre suínos e humanos por água contaminada e carne mal-cozida. A distribuição atual e impacto da infecção pelo HEV na produção suína são desconhecidos. O girovírus aviário tipo 2 (AGV-2) é um novo girovírus, família Circoviridae, identificado no sangue de galinhas com suspeita de infecção pelo vírus da anemia aviária (CAV). O AGV-2 é estreitamente relacionado ao CAV, mas apresenta diferenças genômicas que justificam sua classificação como espécie viral distinta. O AGV-2 parece estar amplamente distribuído em galinhas no Brasil e também em outros países, mas seu potencial patogênico ainda é desconhecido. Finalmente, a longa e intensiva busca por vírus animais relacionados ao vírus da hepatite C humana (HCV) tem levado a identificação de "novos" pestivírus em cães (canine hepacivirus [CHV]), equinos (hepacivirus de não-primatas [NPHV] ou vírus associado à doença de Theiler [TDAV]) e em roedores. Para estes, uma associação clara e definitiva com doença ainda não foi demonstrada e apenas tempo e investigação irão dizer se são patógenos reais ou apenas espectadores.

The list of animal viruses has been frequently added of new members raising permanent concerns to virologists and veterinarians. The pathogenic potential and association with disease have been clearly demonstrated for some, but not for all of these emerging viruses. This review describes recent discoveries of animal viruses and their potential relevance for veterinary practice. Dogs were considered refractory to influenza viruses until 2004, when an influenza A virus subtype H3N8 was transmitted from horses and produced severe respiratory disease in racing greyhounds in Florida/USA. The novel virus, named canine influenza virus (CIV), is considered now a separate virus lineage and has spread among urban canine population in the USA. A new pestivirus (Flaviviridae), tentatively called HoBi-like pestivirus, was identified in 2004 in commercial fetal bovine serum from Brazil. Hobi-like viruses are genetically and antigenically related to bovine viral diarrhea virus (BVDV) and induce similar clinical manifestations. These novel viruses seem to be widespread in Brazilian herds and have also been detected in Southeast Asia and Europe. In 2011, a novel mosquito-borne orthobunyavirus, named Schmallenberg virus (SBV), was associated with fever, drop in milk production, abortion and newborn malformation in cattle and sheep in Germany. Subsequently, the virus disseminated over several European countries and currently represents a real treat for animal health. The origin of SBV is still a matter of debate but it may be a reassortant from previous known bunyaviruses Shamonda and Satuperi. Hepatitis E virus (HEV, family Hepeviridae) is a long known agent of human acute hepatitis and in 1997 was first identified in pigs. Current data indicates that swine HEV is spread worldwide, mainly associated with subclinical infection. Two of the four HEV genotypes are zoonotic and may be transmitted between swine and human by contaminated water and undercooked pork meat. The current distribution and impact of HEV infection in swine production are largely unknown. Avian gyrovirus type 2 (AGV2) is a newly described Gyrovirus, family Circoviridae, which was unexpectedly found in sera of poultry suspected to be infected with chicken anemia virus (CAV). AGV2 is closely related to CAV but displays sufficient genomic differences to be classified as a distinct species. AGV2 seems to be distributed in Brazil and also in other countries but its pathogenic role for chickens is still under investigation. Finally, the long time and intensive search for animal relatives of human hepatitis C virus (HCV) has led to the identification of novel hepaciviruses in dogs (canine hepacivirus [CHV]), horses (non-primate hepaciviruses [NPHV] or Theiler's disease associated virus [TDAV]) and rodents. For these, a clear and definitive association with disease is still lacking and only time and investigation will tell whether they are real disease agents or simple spectators.