Lessons from a Multilaboratorial Task Force for Diagnosis of a Fatal Toxoplasmosis Outbreak in Captive Primates in Brazil.

Toxoplasmosis is an important zoonotic disease caused by the parasite Toxoplasma gondii and is especially fatal for neotropical primates. In Brazil, the Ministry of Health is responsible for national epizootic surveillance, but some diseases are still neglected. Here, we present an integrated investigation of an outbreak that occurred during the first year of the COVID-19 pandemic among eleven neotropical primates housed at a primatology center in Brazil. After presenting non-specific clinical signs, all animals died within four days. A wide range of pathogens were evaluated, and we successfully identified T. gondii as the causative agent within four days after necropsies. The liver was the most affected organ, presenting hemorrhage and hepatocellular necrosis. Tachyzoites and bradyzoite cysts were observed in histological examinations and immunohistochemistry in different organs; in addition, parasitic DNA was detected through PCR in blood samples from all specimens evaluated. A high prevalence of Escherichia coli was also observed, indicating sepsis. This case highlights some of the obstacles faced by the current Brazilian surveillance system. A diagnosis was obtained through the integrated action of researchers since investigation for toxoplasmosis is currently absent in national guidelines. An interdisciplinary investigation could be a possible model for future epizootic investigations in animals.

Seroprevalence of Hantavirus among Manual Cane Cutters and Epidemiological Aspects of HPS in Central Brazil.

Hantavirus pulmonary syndrome (HPS) is a rodent-borne zoonotic disease that is endemic throughout the Americas. Agricultural activities increase exposure to wild rodents, especially for sugarcane cutters. We carried out a survey of the epidemiological aspects of HPS and investigated the prevalence of hantavirus infection in the sugarcane cutter population from different localities in the Brazilian Midwest region. We conducted a retrospective study of all confirmed HPS cases in the state of Goiás reported to the National HPS surveillance system between 2007 and 2017, along with a seroepidemiological study in a population of sugarcane cutters working in Goiás state in 2016, using the anti-hantavirus (Andes) ELISA IgG. A total of 634 serum samples from cane cutters were tested for hantavirus antibodies, with 44 (6.9%) being IgG-reactive according to ELISA. The destination of garbage was the only statistically significant variable (p = 0.03) related to the detection of hantavirus IgG (p < 0.05). We described the epidemiological profile of reported hantavirus cases in Goiás-a highly endemic area for HPS, and where the seroepidemiological study was conducted. Our results increase our knowledge about hantavirus infections in Brazil and highlight the vulnerability of sugarcane cutters to a highly lethal disease that, to date, has no specific treatment or vaccination.

Case Report: Hantavirus Cardiopulmonary Syndrome Diagnostic in the Face of the COVID-19 Pandemic.

Hantavirus cardiopulmonary syndrome (HCPS) is an emerging rodent-borne disease in the Americas. The most common initial symptoms of HCPS are similar to those of COVID-19 and other respiratory infections that evolve rapidly to respiratory failure, resulting from pulmonary edema and shock in about 40% of cases. We describe a fatal case of HCPS in a 24-year-old man who was hospitalized with fever, hemoconcentration, thrombocytopenia, leukocytosis, dry cough and a bilateral diffuse alveolar pulmonary infiltrate during the emergence of the COVID-19 pandemic in Brazil. HCPS needs to be ruled out in patients with clinical manifestations compatible with respiratory infections such as influenza and COVID-19.

Orthohantavirus Survey in Indigenous Lands in a Savannah-Like Biome, Brazil.

In Brazil, the first confirmed cases of hantavirus cardiopulmonary syndrome in Indigenous populations occurred in 2001. The purpose of this study was to determine the seroprevalence of orthohantavirus infections in the Utiariti Indigenous land located in the southeastern region of the Brazilian Amazon. In December 2014 and 2015, a survey was conducted using an enzyme-linked immunosorbent assay in nine villages belonging to the Haliti-Paresí Indigenous communities. A total of 301 participants were enrolled in the study. Of the two study cohorts, the one from 2014 showed a prevalence of 12.4%, whereas the one from 2015 had a serum prevalence of 13.4%. Analysis of the paired samples of 110 Indigenous people who participated in both stages of the study enabled identification of four individuals who had seroconverted during the study period. Identifying the circulation of orthohantaviruses in the Utiariti Indigenous land highlights a serious public health problem in viral expansion and highlights the need to implement preventive measures appropriate to the sociocultural reality of these communities.

The Serological Cross-Detection of Bat-Borne Hantaviruses: A Valid Strategy or Taking Chances?

Bats are hosts of a range of viruses, and their great diversity and unique characteristics that distinguish them from all other mammals have been related to the maintenance, evolution, and dissemination of these pathogens. Recently, very divergent hantaviruses have been discovered in distinct species of bats worldwide, but their association with human disease remains unclear. Considering the low success rates of detecting hantavirus RNA in bat tissues and that to date no hantaviruses have been isolated from bat samples, immunodiagnostic tools could be very helpful to understand pathogenesis, epidemiology, and geographic range of bat-borne hantaviruses. In this sense, we aimed to identify in silico immunogenic B-cell epitopes present on bat-borne hantaviruses nucleoprotein (NP) and verify if they are conserved among them and other selected members of Mammantavirinae, using a combination of (the three most used) different prediction algorithms, ELLIPRO, Discotope 2.0, and PEPITO server. To support our data, we in silico modeled 3D structures of NPs from representative members of bat-borne hantaviruses, using comparative and ab initio methods due to the absence of crystallographic structures of studied proteins or similar models in the Protein Data Bank. Our analysis demonstrated the antigenic complexity of the bat-borne hantaviruses group, showing a low sequence conservation of epitopes among members of its own group and a minor conservation degree in comparison to Orthohantavirus, with a recognized importance to public health. Our data suggest that the use of recombinant rodent-borne hantavirus NPs to cross-detect antibodies against bat- or shrew-borne viruses could underestimate the real impact of this virus in nature.

Padronização de procedimentos higienicossanitários de restaurantes de culinária japonesa / Standardization of hygienic-sanitary procedures in japanese cuisine restaurants

O presente estudo objetivou elaborar procedimentos higienicossanitários padronizados voltados especificamente para restaurantes de culinária japonesa. Para tal, foi conduzido um estudo de caso, realizado entre maio e junho de 2019, em um restaurante típico da culinária japonesa em Joinville ­ SC. O instrumento de coleta foi uma lista de verificação em boas práticas baseada na legislação sanitária brasileira vigente, especialmente a resolução RDC nº 216/2004 da Agência Nacional de Vigilância Sanitária, contendo 31 itens quanto à matéria-prima e preparação dos alimentos; armazenamento e exposição ao consumo do alimento; higiene dos utensílios e ambiente; higiene do manipulador. Dos 31 itens avaliados, verificou-se 25,8% de inconformidades no local estudado, sendo estes, em sua maioria, relacionados ao preparado do arroz temperado (gohan). Porém, a maioria dos itens estava de acordo com as recomendações ou não se aplicavam ao local. Com base nas informações obtidas com o estudo de caso, foram analisados os pontos críticos que deveriam ser destacados e então, elaborou-se um quadro com os procedimentos que devem ser padronizados, especificamente para restaurantes de cozinha japonesa, quanto aos mesmos critérios avaliados: matéria-prima e preparação dos alimentos; exposição ao consumo do alimento; higiene dos utensílios e ambiente; higiene do manipulador, a fim de garantir a qualidade higienicossanitária dos alimentos preparados. O material desenvolvido poderá ser afixado nas cozinhas para guiar os manipuladores e ser utilizado em treinamentos periódicos. Assim, o quadro de procedimentos higienicossanitários padronizados em restaurantes japoneses pode ser uma ferramenta útil para auxiliar na garantia da segurança dos alimentos, por meio das boas práticas na manipulação de alimentos

The present study aimed to developed standardized hygienic-sanitary procedures specifically for Japanese cuisine restaurants. Thus, a case study was conducted, conducted between May and June 2019, in a traditional Japanese cuisine restaurant in Joinville ­ SC. The data collection instrument was a checklist on good practices based on current Brazilian sanitary legislation, especially Resolution RDC No. 216/2004 of the National Health Surveillance Agency, containing 31 items regarding raw material and food preparation; food storage and consumption exposure; hygiene of utensils and environment; handler hygiene. Of the 31 items analyzed, 25.8% of non-conformities were found in the study, most of them related to the preparation of seasoned rice (gohan). However, most items were in accordance with the recommendations or did not apply to the restaurant. Based on the information obtained from the case study, the critical points that should be highlighted were analyzed and a table was prepared with the procedures that should be standardized, specifically for Japanese cuisine restaurants, regarding the same criteria analyzed: raw materials and food preparation; exposure to food consumption; hygiene of utensils and environment; hygiene of the handler in order to guarantee the hygienic-sanitary quality of the prepared foods. The developed material can be posted in the kitchens to guide the handlers and be used in periodic training. Thus, standardized hygienic-sanitary procedures in Japanese restaurants can be a useful tool to help ensure food safety through good food handling practices

Padronização de procedimentos higienicossanitários de restaurantes de culinária japonesa / Standardization of hygienic-sanitary procedures in Japanese cuisine restaurants

O presente estudo objetivou elaborar procedimentos higienicossanitários padronizados voltados especificamente para restaurantes de culinária japonesa. Para tal, foi conduzido um estudo de caso, realizado entre maio e junho de 2019, em um restaurante típico da culinária japonesa em Joinville – SC. O instrumento de coleta foi uma lista de verificação em boas práticas baseada na legislação sanitária brasileira vigente, especialmente a resolução RDC nº 216/2004 da Agência Nacional de Vigilância Sanitária, contendo 31 itens quanto à matéria-prima e preparação dos alimentos; armazenamento e exposição ao consumo do alimento; higiene dos utensílios e ambiente; higiene do manipulador. Dos 31 itens avaliados, verificou-se 25,8% de inconformidades no local estudado, sendo estes, em sua maioria, relacionados ao preparado do arroz temperado (gohan). Porém, a maioria dos itens estava de acordo com as recomendações ou não se aplicavam ao local. Com base nas informações obtidas com o estudo de caso, foram analisados os pontos críticos que deveriam ser destacados e então, elaborou-se um quadro com os procedimentos que devem ser padronizados, especificamente para restaurantes de cozinha japonesa, quanto aos mesmos critérios avaliados: matéria-prima e preparação dos alimentos; exposição ao consumo do alimento; higiene dos utensílios e ambiente; higiene do manipulador, a fim de garantir a qualidade higienicossanitária dos alimentos preparados. O material desenvolvido poderá ser afixado nas cozinhas para guiar os manipuladores e ser utilizado em treinamentos periódicos. Assim, o quadro de procedimentos higienicossanitários padronizados em restaurantes japoneses pode ser uma ferramenta útil para auxiliar na garantia da segurança dos alimentos, por meio das boas práticas na manipulação de alimentos.

The present study aimed to developed standardized hygienic-sanitary procedures specifically for Japanese cuisine restaurants. Thus, a case study was conducted, conducted between May and June 2019, in a traditional Japanese cuisine restaurant in Joinville – SC. The data collection instrument was a checklist on good practices based on current Brazilian sanitary legislation, especially Resolution RDC No. 216/2004 of the National Health Surveillance Agency, containing 31 items regarding raw material and food preparation; food storage and consumption exposure; hygiene of utensils and environment; handler hygiene. Of the 31 items analyzed, 25.8% of non-conformities were found in the study, most of them related to the preparation of seasoned rice (gohan). However, most items were in accordance with the recommendations or did not apply to the restaurant. Based on the information obtained from the case study, the critical points that should be highlighted were analyzed and a table was prepared with the procedures that should be standardized, specifically for Japanese cuisine restaurants, regarding the same criteria analyzed: raw materials and food preparation; exposure to food consumption; hygiene of utensils and environment; hygiene of the handler in order to guarantee the hygienic-sanitary quality of the prepared foods. The developed material can be posted in the kitchens to guide the handlers and be used in periodic training. Thus, standardized hygienic-sanitary procedures in Japanese restaurants can be a useful tool to help ensure food safety through good food handling practices.

A Retrospective Survey of Rodent-borne Viruses in Rural Populations of Brazilian Amazon.

INTRODUCTION: The Amazon tropical rainforest has the most dense and diverse ecosystem worldwide. A few studies have addressed rodent-borne diseases as potential hazards to humans in this region. METHODS: A retrospective survey was conducted using enzyme-linked immunosorbent assay for detecting mammarenavirus and orthohantavirus antibodies in 206 samples collected from rural settlers of the Brazilian Western Amazonian region. RESULTS: Six (2.91%) individuals in the age group of 16 to 36 years were found to possess antibodies against mammarenavirus. CONCLUSION: Evidence of previous exposure to mammarenavirus in the rural population points to its silent circulation in this region.

Avaliação das boas práticas em restaurantes do município de Joinville, Santa Catarina / Evaluation of good practices in restaurants in Joinville, Santa Catarina, Brazil

Introdução: O aumento da alimentação fora de casa tem relação direta com a expansão dos serviços de alimentação. Por conta dessa expansão, tornam-se necessários estudos que avaliem as condições higiênico-sanitárias e de boas práticas nos estabelecimentos. Objetivo: Avaliar as boas práticas de manipulação de alimentos em restaurantes de Joinville-SC. Método: Coleta de dados realizada por meio da aplicação da lista de verificação de boas práticas para serviços de alimentação (checklist), integrante da Resolução Normativa nº 3 da Diretoria de Vigilância Sanitária do Estado de Santa Catarina, em conformidade com a RDC n° 216/2004 da Agência Nacional de Vigilância Sanitária. Resultados: Aplicou-se a checklist em 30 restaurantes, sendo que apenas um foi classificado como plenamente satisfatório, com índice de adequação de 81,8%; dois foram classificados como satisfatórios, com média de 53,0% de conformidades; e os demais 27 estabelecimentos obtiveram índice de adequação inferior a 50,0%, sendo classificados como insatisfatórios. Conclusão: Considerando os resultados, se faz necessário adotar medidas operacionais e corretivas para controle dos pontos críticos. Estas devem ser realizadas e reforçadas pelos nutricionistas que atuam em restaurantes, com o objetivo de reduzir as não conformidades e, consequentemente, os riscos de doenças transmitidas por alimentos. (AU)

Introduction: The increase in eating out of home is directly related to the expansion of food service. Because of this expansion, studies are needed to evaluate hygienicsanitary conditions and good practices in food services. Objective: Evaluate good food handling practices in restaurants in Joinville-SC, Brazil. Method: Data collection performed through the application of good practice checklist for food service, part of the Normative Resolution No. 3 of the Directorate of Sanitary Surveillance of Santa Catarina State, Brazil, in accordance with RDC No. 216 / 2004 of the Brazilian Health Surveillance Agency. Results: Checklist was applied to 30 restaurants, only one was classified as fully satisfactory, with an adequacy index of 81.8%; two were classified as satisfactory, with an average of 53.0% adequacy; and the remaining 27 restaurants obtained an adequacy index below 50.0%, being classified as unsatisfactory. Conclusion: Considering the results, it is necessary to adopt operational and corrective measures to control the critical points. These must be carried out and reinforced by nutritionists who work in restaurants, aiming to reduce non-conformities and, consequently, the risks of foodborne diseases. (AU)

A Retrospective Survey of Rodent-borne Viruses in Rural Populations of Brazilian Amazon

Abstract INTRODUCTION: The Amazon tropical rainforest has the most dense and diverse ecosystem worldwide. A few studies have addressed rodent-borne diseases as potential hazards to humans in this region. METHODS: A retrospective survey was conducted using enzyme-linked immunosorbent assay for detecting mammarenavirus and orthohantavirus antibodies in 206 samples collected from rural settlers of the Brazilian Western Amazonian region. RESULTS: Six (2.91%) individuals in the age group of 16 to 36 years were found to possess antibodies against mammarenavirus. CONCLUSION: Evidence of previous exposure to mammarenavirus in the rural population points to its silent circulation in this region.

Identification and validation of specific B-cell epitopes of hantaviruses associated to hemorrhagic fever and renal syndrome.

BACKGROUND: Orthohantavirus infection is a neglected global health problem affecting approximately 200,000 people/year, spread by rodent hosts and associated to fatal human diseases, such as hemorrhagic fever with renal syndrome (HFRS) and orthohantavirus cardiopulmonary syndrome (HCPS). Circulation of HFRS-associated orthohantaviruses, such as Seoul, Gou, Amur, Dobrava and Hantaan, are supposed to be restricted to Eurasian countries even though their hosts can be a worldwide distribution. Few confirmed HFRS orthohantavirus infections in humans have been reported in American countries, but due to lower medical awareness of the symptoms of this zoonosis, it could be associated to viral underreporting or to misdiagnosis with several tropical hemorrhagic diseases. Serological evidence of orthohantavirus infections, using enzyme-linked immunosorbent assay for the presence of immunoglobulin M and G against recombinant nucleoprotein protein, remains as an essential assay for viral surveillance. In this study, we aimed to identify in silico immunogenic B-cell linear epitopes present on orthohantavirus nucleoprotein that are exclusive to HFRS-related species. METHODOLOGY/PRINCIPAL FINDINGS: In silico analysis were performed using Seoul orthohantavirus nucleoprotein (SHNP) sequence as a model. Linear B-cell-epitopes on SHNP and its immunogenicity were predicted by BepiPred-2.0 and Vaxijen algorithms, respectively. The conservancy of predicted epitopes was compared with the most clinically relevant HFRS or HCPS-associated orthohantavirus, aiming to identify specific sequences from HFRS-orthohantavirus. Peptide validation was carried out by ELISA using Balb/c mice sera immunized with purified recombinant rSHNP. Peptides cross-reactivity against HCPS orthohantavirus were evaluated using immunized sera from mice injected with recombinant Juquitiba orthohantavirus nucleoprotein (rJHNP). CONCLUSION/SIGNIFICANCE: In silico analysis revealed nine potential immunogenic linear B-cell epitopes from SHNP; among them, SHNP(G72-D110) and SHNP(P251-D264) showed a high degree of sequence conservation among HFRS-related orthohantavirus and were experimentally validated against rSHNP-IMS and negatively validated against rJHNP-IMS. Taken together, we identified and validated two potential antigenic B-cell epitopes on SHNP, which were conserved among HFRS-associated orthohantavirus and could be applied to the development of novel immunodiagnostic tools for orthohantavirus surveillance.

Hantavirus pulmonary syndrome in children: case report and case series from an endemic area of Brazil.

Hantavirus Cardiopulmonary Syndrome (HCPS) is an important emergent zoonosis associated with wild rodents in Brazil, where this viral infection in children is generally rare. We present HCPS in a child from the Pantanal Biome and a review of all reported pediatric cases in Mato Grosso State, an endemic area for HCPS in Brazil. The investigation used the Information System for Notifiable Diseases database (SINAN). A 12-year-old boy was hospitalized with fever and respiratory failure and hantavirus IgM and IgG antibodies were detected by ELISA in serum samples. During the period of 1999 to 2016, 32 HCPS pediatric cases confirmed by serology were reported to SINAN with a mortality rate of 34.4%. The possibility of hantavirus infection in children with acute febrile illness associated with respiratory failure should be considered mainly in recognized endemic areas as Mato Grosso State, contradicting a hypothesis that children are more protected from lung involvement.

Seroprevalence of rodent-borne viruses in Afro-descendent communities in Brazil.

During the Brazilian slavery period, many African migrants were brought to the American continent. Historically, some of these migrants escaped from the Brazilian gold mines and farms to which they had been brought and settled in remote valleys and this was the main mode of resistance to the slavery system. These runaway-slave descendant communities are called quilombos, a group with distinct ethnic identity, specific behavioral habits, including geographic isolation and conservative practices. The objective of this study was to investigate the prevalence of rodent-borne viruses in two Afro-descendent communities from Mato Grosso do Sul State, Midwestern Brazil. A total of 319 individuals from rural and urban quilombola communities were enrolled. Twelve (3.76%) had anti-rodent-borne virus IgG antibodies. Seven (2.19%) were anti-mammarenavirus reactive and nine (2.82%) had anti-orthohantavirus antibodies. The literature includes limited data on the health status of quilombola communities, but all the studies emphasize the disparity of attention of local healthcare personnel to these communities compared to the general population. The findings of this study highlight the vulnerability and the precarious health conditions of quilombola groups, especially those living in rural areas and thus, point to the need of preventive measures to improve access to healthcare for this ethnic group.

Aporé virus, a novel mammarenavirus (Bunyavirales: Arenaviridae) related to highly pathogenic virus from South America.

Here, we report the complete genome sequence of the Aporé virus (Bunyavirales: Arenaviridae), obtained from a wild rodent Oligoryzomys mattogrossae captured in Mato Grosso do Sul state, Brazil. The genome of this virus showed strong similarity to highly pathogenic mammarenavirus from South America.

Malaria and Hantavirus Pulmonary Syndrome in Gold Mining in the Amazon Region, Brazil.

People living in mining regions are exposed to numerous biological agents by several specific types of transmission mechanisms. This study is designed to describe fatal hantavirus pulmonary syndrome (HPS) cases confirmed by serology and molecular analysis, where a seroprevalence survey was conducted in the gold mining regions of the state of Mato Grosso, in the official Amazon region, Brazil. Two fatal cases of HPS were confirmed in a mining area in the Legal Amazon, where malaria is one of the most important public health problems. A molecular analysis detected the presence of the genome of the Castelo dos Sonhos virus. Out of the 112 blood samples analyzed, five were positive for Plasmodium infection (four P. falciparum and one P. vivax), and four were seropositive for hantavirus, showing a seroprevalence of 3.57%. One of the four miners who was seroreactive for hantavirus concomitantly had P. falciparum infection, which was confirmed by thick blood smear. This manuscript highlights the importance of considering hantavirus pulmonary syndrome as a diagnostic possibility in febrile infection associated with pulmonary manifestations in mining areas where malaria cases are often identified.

The mystery of the phylogeographic structural pattern in rodent-borne hantaviruses.

Hantaviruses (order Bunyavirales, family Hantaviridae) are important zoonotic pathogens. Because of the great diversity of their reservoir hosts, hantaviruses are excellent models to evaluate the dynamics of virus-host co-evolution. To understand the mechanisms behind the evolutionary history of hantaviruses through virus-reservoir interactions, it is important to know how the radiation and diversity of hantaviruses occurred. In this paper, we evaluate the pattern of hantavirus diversification based on a complete S segment representing major groups of hantaviruses found in the Americas. Phylogenetic analyses revealed a high degree of phylogeographic structure and a surprising pattern of geographical distribution of New World hantaviruses. The available data suggest that hantaviruses related to the Arvicolinae rodent subfamily in North America probably emerged and initially adapted from a shared common ancestor of the Tula virus. The first clade of hantaviruses associated with Neotominae occupied a stem lineage, especially those that emerged in Central America or Mexico. Hantaviruses from Central America and Mexico found in Neotominae rodents spread northward and probably gave rise to the first phylogroup of hantaviruses associated with Sigmodontinae in North America. Two preferential host-switching transmissions in hantaviruses apparently gave rise to two different paraphyletic group in Neotominae and Sigmodontinae. Our study supports a probable epicenter of diversification in Central America and/or Mexico for hantaviruses related to both the Neotominae and Sigmodontinae subfamilies.

Aporé virus, a novel mammarenavirus (Bunyavirales: Arenaviridae) related to highly pathogenic virus from South America

Here, we report the complete genome sequence of the Aporé virus (Bunyavirales: Arenaviridae), obtained from a wild rodent Oligoryzomys mattogrossae captured in Mato Grosso do Sul state, Brazil. The genome of this virus showed strong similarity to highly pathogenic mammarenavirus from South America.

Rodent-borne viruses survey in rural settlers from Central Brazil.

Anthropogenic environmental changes arising from settlement and agriculture include deforestation and replacement of natural vegetation by crops providing opportunities for pathogen spillover from animals to humans. This study aimed to investigate the prevalence of rodent-borne virus infections in seven rural settlements from Midwestern Brazil. Of the 466 individuals tested 12 (2.57%) were reactive for orthohantavirus and 3 (0.64%) for mammarenavirus. These rural settlers lived under unfavorable infrastructure, socioeconomic disadvantages, and unsanitary conditions, representing a risk for rodent-borne infections. Development of public policies towards the improvement of health, sanitation and awareness of rodent-borne diseases in improvised camps and settlements is imperative, in order to reduce morbidity and mortality caused by these diseases.