Patterns and spatial distribution of sea turtle strandings in Alagoas, Brazil.

This study makes a descriptive analysis of necropsied sea turtles registered in the Biota Conservation Institute database between May 2018 and May 2022 on the coast of Alagoas, Brazil. During this period, 79 animals of four species were necropsied: 87.4 % (69) Chelonia mydas, 6.3 % (5) Caretta caretta, 3.8 % (3) Lepidochelys olivacea and 2.5 % (2) Eretmochelys imbricata. C. mydas was the most frequent species, mainly juvenile females. In 29.1 % (23/79) evidence of anthropogenic interactions was found (e.g., fishing net marks, plastic waste in the digestive tract, trauma from collisions with boats). Cutaneous tumors suggestive of fibropapillomatosis in 35.4 % (28/79), in C. mydas and E. imbricata, half were in an area of high eutrophication, close to the capital. Endoparasites were found in 46.8 % (37/79) individuals. Information on strandings in the region is essential for understanding the use of the area and the impacts to which these animals are exposed.

Wild mammals involved in the transmission of Trypanosoma cruzi and food sources of Triatoma sherlocki in an endemic region of northeastern Brazil.

The present study, carried out in the municipality of Gentio do Ouro, Bahia, Brazil aimed to evaluate which wild mammals may be involved in the transmission of T. cruzi and which are the blood sources for triatomines collected in the study area. PCR analysis of 31 wild mammals captured revealed T. cruzi infection in 6.4% (2/31): one specimen of the opossum Didelphis albiventris (1/3) and one of the rodent Kerodon rupestris (1/5); despite being more frequent in the area, no specimen of the rodent Thrichomys sp. (0/23) was infected. A total of 169 triatomines were captured. The conclusive detection of food sources was possible only for Triatoma sherlocki Papa et al., 2002 (n = 56), with evidence for: K. rupestris (35.7%), Gallus (17.9%), D. albiventris (14.3%), Homo sapiens (14.3%), Tropidurus hispidus (7.1%), Leopardus geoffroyi (5.3%), Conepatus semistriatus (1.8%), Thrichomys inermis (1.8%) and Rattus norvegicus (1.8%). Triatomines of the species T. sherlocki showed food eclecticism, including feeding on humans, with some of them being captured at dwellings. These facts make this triatomine a potential link for the transmission of T. cruzi between wild and anthropic environments, highlighting a latent risk of the reemergence of Chagas disease outbreaks.

Frequent Infection of Cats With SARS-CoV-2 Irrespective of Pre-Existing Enzootic Coronavirus Immunity, Brazil 2020.

Carnivores such as cats and minks are highly susceptible to SARS-CoV-2. Brazil is a global COVID-19 hot spot and several cases of human-to-cat transmission have been documented. We investigated the spread of SARS-CoV-2 by testing 547 domestic cats sampled between July-November 2020 from seven states in southern, southeastern, and northeastern Brazil. Moreover, we investigated whether immune responses elicited by enzootic coronaviruses affect SARS-CoV-2 infection in cats. We found infection with significantly higher neutralizing antibody titers against the Gamma variant of concern, endemic in Brazil during 2020, than against an early SARS-CoV-2 B.1 isolate (p<0.0001), validating the use of Gamma for further testing. The overall SARS-CoV-2 seroprevalence in Brazilian cats during late 2020 validated by plaque reduction neutralization test (PRNT90) was 7.3% (95% CI, 5.3-9.8). There was no significant difference in SARS-CoV-2 seroprevalence in cats between Brazilian states, suggesting homogeneous infection levels ranging from 4.6% (95% CI, 2.2-8.4) to 11.4% (95% CI, 6.7-17.4; p=0.4438). Seroprevalence of the prototypic cat coronavirus Feline coronavirus (FCoV) in a PRNT90 was high at 33.3% (95% CI, 24.9-42.5) and seroprevalence of Bovine coronavirus (BCoV) was low at 1.7% (95% CI, 0.2-5.9) in a PRNT90. Neutralizing antibody titers were significantly lower for FCoV than for SARS-CoV-2 (p=0.0001), consistent with relatively more recent infection of cats with SARS-CoV-2. Neither the magnitude of SARS-CoV-2 antibody titers (p=0.6390), nor SARS-CoV-2 infection status were affected by FCoV serostatus (p=0.8863). Our data suggest that pre-existing immunity against enzootic coronaviruses neither prevents, nor enhances SARS-CoV-2 infection in cats. High SARS-CoV-2 seroprevalence already during the first year of the pandemic substantiates frequent infection of domestic cats and raises concerns on potential SARS-CoV-2 mutations escaping human immunity upon spillback.

Evidence against Zika virus infection of pets and peri-domestic animals in Latin America and Africa.

Decades after its discovery in East Africa, Zika virus (ZIKV) emerged in Brazil in 2013 and infected millions of people during intense urban transmission. Whether vertebrates other than humans are involved in ZIKV transmission cycles remained unclear. Here, we investigate the role of different animals as ZIKV reservoirs by testing 1723 sera of pets, peri-domestic animals and African non-human primates (NHP) sampled during 2013-2018 in Brazil and 2006-2016 in Côte d'Ivoire. Exhaustive neutralization testing substantiated co-circulation of multiple flaviviruses and failed to confirm ZIKV infection in pets or peri-domestic animals in Côte d'Ivoire (n=259) and Brazil (n=1416). In contrast, ZIKV seroprevalence was 22.2% (2/9, 95% CI, 2.8-60.1) in West African chimpanzees (Pan troglodytes verus) and 11.1% (1/9, 95% CI, 0.3-48.3) in king colobus (Colobus polycomos). Our results indicate that while NHP may represent ZIKV reservoirs in Africa, pets or peri-domestic animals likely do not play a role in ZIKV transmission cycles.

Eco-epidemiology of vectorial Trypanosoma cruzi transmission in a region of northeast Brazil.

Chagas disease (CD) is a parasitic zoonosis endemic in Brazil. Despite virtual control of Triatoma infestans, the main domesticated vector of Trypanosoma cruzi, vectorial transmission by other triatomine species persists in some rural communities. This study aims to characterize triatomines role in transmitting T. cruzi to dogs and humans in the district of Santo Inácio, located in the northwest region of the state of Bahia, Brazil. It also describes environmental factors in housings associated with insect occurrence and assesses the perception, knowledge, and preventive practices adopted by the population regarding CD. Blood samples of humans and dogs, and biological samples of triatomines, were collected between November 2018 and February 2019 and subjected to the detection of T. cruzi by serological and molecular biology tests. Also, we applied a questionnaire to research the perception, knowledge, and local practices of people related to CD. The capture of triatomines in households was associated with exploratory variables of the questionnaires using multivariate logistic regression (p < 0.05). The 155 triatomines captured in the wild and domestic environment were of the species Triatoma sherlocki (n = 151), Panstrongylus sherlocki (n = 1) and Triatoma sordida (n = 3), and had a natural infection rate for T. cruzi by PCR of 18.5%, 100% and 0%, respectively. District residents (n = 126) were seronegative for T. cruzi, while 17.5% (7/40) of the dogs were seropositive. The fact that residents are aware that triatomines can "cause" CD was configured as a protection factor for residents according to the fitted logistic regression model (p = 0.04). However, respondents have limited perception and knowledge about the CD, prevention and control practices for triatomines in a household. The results suggest the existence of a domestic cycle of transmission of T. cruzi between triatomines and dogs, configuring a latent risk of infection to the human population of Santo Inácio. Studies that clarify the potential for the establishing of intrusive triatomines in households, surveillance actions for triatomines, and health education in rural communities are indispensable to prevent the reemergence of CD in vulnerable regions of Brazil and other American countries with similar epidemiological characteristics.

Ecologic Determinants of West Nile Virus Seroprevalence among Equids, Brazil.

Among 713 equids sampled in northeastern Brazil during 2013-2018, West Nile virus seroprevalence was 4.5% (95% CI 3.1%-6.3%). Mathematical modeling substantiated higher seroprevalence adjacent to an avian migratory route and in areas characterized by forest loss, implying increased risk for zoonotic infections in disturbed areas.

Identification of animal hosts of Fort Sherman virus, a New World zoonotic orthobunyavirus.

An orthobunyavirus termed Fort Sherman virus (FSV) was isolated in 1985 from a febrile US soldier in Panama, yet potential animal reservoirs remained unknown. We investigated sera from 192 clinically healthy peri-domestic animals sampled in northeastern Brazil during 2014-2018 by broadly reactive RT-PCR for orthobunyavirus RNA, including 50 cattle, 57 sheep, 35 goats and 50 horses. One horse sampled in 2018 was positive (0.5%; 95% CI, 0.01-3.2) at 6.2 × 103 viral RNA copies/mL. Genomic comparisons following virus isolation in Vero cells and deep sequencing revealed high identity of translated amino acid sequences between the new orthobunyavirus and the Panamanian FSV prototype (genes: L, 98.8%; M, 83.5%; S, 100%), suggesting these viruses are conspecific. Database comparisons revealed even higher genomic identity between the Brazilian FSV and taxonomically unassigned Argentinian mosquito- and horse-derived viruses sampled in 1965, 1982 and 2013 with only 1.1% maximum translated amino acid distances across viral genes, suggesting the Argentinian viruses were also distinct FSV strains. The Panamanian FSV strain was an M gene reassortant relative to all Southern American FSV strains, clustering phylogenetically with Cache Valley virus (CVV). Mean dN/dS ratios among FSV genes ranged from 0.03 to 0.07, compatible with strong purifying selection. FSV-specific neutralizing antibodies occurred at relatively high end-point titres in the range of 1:300 in 22.0% of horses (11 out of 50 animals), 8.0% of cattle (4/50 animals), 7.0% of sheep (4/57 animals) and 2.9% of goats (1/35 animals). High specificity of serologic testing was suggested by significantly higher overall FSV-specific compared to CVV- and Bunyamwera virus-specific end-point titres (p = .009), corroborating a broad vertebrate host range within peri-domestic animals. Growth kinetics using mosquito-, midge- and sandfly-derived cell lines suggested Aedes mosquitos as potential vectors. Our findings highlight the occurrence of FSV across a geographic range exceeding 7,000 km, surprising genomic conservation across a time span exceeding 50 years, M gene-based reassortment events, and the existence of multiple animal hosts of FSV.

Seroepidemiological survey of canine visceral leishmaniasis in a continental island of northeast Brazil / Investigação soroepidemiológica da leishmaniose visceral canina em uma ilha continental do Nordeste do Brasil

Introduction: Visceral leishmaniasis (VL) is endemic disease in the neighboring municipalities of the continental island Ilha de Maré, Salvador, Bahia, Brazil. Nevertheless, VL has not been reported in the island itself. Objective: the present study aimed to investigate the seroprevalence and clinical signs of Canine Visceral Leishmaniasis (CVL) and to identify the sand fly population present in the village of Botelho, Ilha de Maré. Methodology: sera of local dogs were tested for anti-Leishmania IgG by immunoassays (screening with TR DPP ™ rapid test and confirmatory with indirect ELISA, Bio-Manguinhos/Fiocruz) and an entomological survey was conducted to estimate and identify the phlebotomine fauna of the region. Results: seven out of 106 samples (6.6%) were positive using rapid test. These positive samples were sent to the Central Laboratory of Bahia for confirmation by indirect ELISA. However, all samples presentednegative results. Nine specimens of Pressatia choti, subfamily Phlebotominae were identified, being this species frequently found in areas with cutaneous leishmaniasis transmission in Brazil. Conclusion: although this work did not confirm the presence of CVL in Ilha de Maré, new serological and entomological studies in a larger area are required for the maintenance of the epidemiological surveillance in the emphasized insular area.

Introdução: Leishmaniose Visceral (LV) é uma doença endêmica em municípios vizinhos à Ilha de Maré, situada na plataforma continental do município de Salvador, Bahia, Brasil. Entretanto, casos de LV não tem sido notificados nesta Ilha. Objetivo: O presente trabalho objetivou investigar a soroprevalência e sinais clínicos de Leishmaniose Visceral Canina (LVC) e identificar a população de flebótomos presentes no povoado de Botelho, Ilha de Maré. Metodologia: soro de cães locais foram testados para IgG anti-Leishmania por imunoensaios (triagem com teste rápido TR DPP™ e confirmatório com Elisa Indireto, Bio-Manguinhos/Fiocruz) e uma investigação entomológica foi conduzida para estimar e identificar a fauna flebotomínea da região. Resultados: sete de 106 amostras (6,6%) foram positivas usando o teste rápido. As amostras positivas foram encaminhadas ao Laboratório Central da Bahia para a confirmação por ELISA indireto. Entretanto, essas amostras apresentaram resultado negativo. Foram encontrados nove exemplares da espécie Pressatia choti, subfamília Phlebotominae, espécie frequentemente encontrada em áreas de transmissão de leishmaniose cutânea no Brasil. Conclusão: apesar deste trabalho não ter confirmado a presença de LVC na Ilha de Maré, novos inquéritos sorológicos e analises entomológicas em uma maior área são necessários para a manutenção de uma vigilância epidemiológica na região insular em destaque.

Knowledge, practice and perception of human-marsupial interactions in health promotion.

INTRODUCTION: Approximately 60% of emerging pathogens originate from wild animals, with mammals being the main hosts. Among Didelphis, which are restricted to the Americas, the species Didelphis aurita and Didelphis albiventris are particularly widely distributed throughout Brazil, where they act as hosts for several pathogens transmissible to humans. The reduction of their natural habitat has resulted in the adaptation of these species to human environments. Animals hunting, due to food necessity or cultural habit, may increase pathogen exposure with a potential to zoonotic disease transmission. METHODOLOGY: From November to December 2016, we administered semi-structured questionnaires in a rural community in northeastern Brazil to assess knowledge, practices and perceptions regarding human-didelphis interactions and possible exposure to zoonoses. RESULTS: There were 213 respondents. Based on photographs of D. albiventris and D. aurita, 91.2% and 78% respondents, respectively, identified the animal by the popular name "sariguê", 61% (130/213) believed the animal could convey any disease, 4.7% stated they did not, and 34% did not know. Opossum meat consumption was reported by 20.2% (43/213), of which 58.1% admitted disease transmission possibility. Only 15.9% of respondents had a secondary or higher education level. The distribution of these frequencies is discussed according to the respondents educational level. CONCLUSIONS: The results reveal the need to carry out health educational activities, including better community knowledge regarding the possible exposure to pathogens due to marsupial consumption.

A Novel Marsupial Hepatitis A Virus Corroborates Complex Evolutionary Patterns Shaping the Genus Hepatovirus.

The discovery of highly diverse nonprimate hepatoviruses illuminated the evolutionary origins of hepatitis A virus (HAV) ancestors in mammals other than primates. Marsupials are ancient mammals that diverged from other Eutheria during the Jurassic. Viruses from marsupials may thus provide important insight into virus evolution. To investigate Hepatovirus macroevolutionary patterns, we sampled 112 opossums in northeastern Brazil. A novel marsupial HAV (MHAV) in the Brazilian common opossum (Didelphis aurita) was detected by nested reverse transcription-PCR (RT-PCR). MHAV concentration in the liver was high, at 2.5 × 109 RNA copies/g, and at least 300-fold higher than those in other solid organs, suggesting hepatotropism. Hepatovirus seroprevalence in D. aurita was 26.6% as determined using an enzyme-linked immunosorbent assay (ELISA). Endpoint titers in confirmatory immunofluorescence assays were high, and marsupial antibodies colocalized with anti-HAV control sera, suggesting specificity of serological detection and considerable antigenic relatedness between HAV and MHAV. MHAV showed all genomic hallmarks defining hepatoviruses, including late-domain motifs likely involved in quasi-envelope acquisition, a predicted C-terminal pX extension of VP1, strong avoidance of CpG dinucleotides, and a type 3 internal ribosomal entry site. Translated polyprotein gene sequence distances of at least 23.7% from other hepatoviruses suggested that MHAV represents a novel Hepatovirus species. Conserved predicted cleavage sites suggested similarities in polyprotein processing between HAV and MHAV. MHAV was nested within rodent hepatoviruses in phylogenetic reconstructions, suggesting an ancestral hepatovirus host switch from rodents into marsupials. Cophylogenetic reconciliations of host and hepatovirus phylogenies confirmed that host-independent macroevolutionary patterns shaped the phylogenetic relationships of extant hepatoviruses. Although marsupials are synanthropic and consumed as wild game in Brazil, HAV community protective immunity may limit the zoonotic potential of MHAV.IMPORTANCE Hepatitis A virus (HAV) is a ubiquitous cause of acute hepatitis in humans. Recent findings revealed the evolutionary origins of HAV and the genus Hepatovirus defined by HAV in mammals other than primates in general and in small mammals in particular. The factors shaping the genealogy of extant hepatoviruses are unclear. We sampled marsupials, one of the most ancient mammalian lineages, and identified a novel marsupial HAV (MHAV). The novel MHAV shared specific features with HAV, including hepatotropism, antigenicity, genome structure, and a common ancestor in phylogenetic reconstructions. Coevolutionary analyses revealed that host-independent evolutionary patterns contributed most to the current phylogeny of hepatoviruses and that MHAV was the most drastic example of a cross-order host switch of any hepatovirus observed so far. The divergence of marsupials from other mammals offers unique opportunities to investigate HAV species barriers and whether mechanisms of HAV immune control are evolutionarily conserved.

A novel hepatitis B virus species discovered in capuchin monkeys sheds new light on the evolution of primate hepadnaviruses.

BACKGROUND & AIMS: All known hepatitis B virus (HBV) genotypes occur in humans and hominoid Old World non-human primates (NHPs). The divergent woolly monkey HBV (WMHBV) forms another orthohepadnavirus species. The evolutionary origins of HBV are unclear. METHODS: We analysed sera from 124 Brazilian monkeys collected during 2012-2016 for hepadnaviruses using molecular and serological tools, and conducted evolutionary analyses. RESULTS: We identified a novel orthohepadnavirus species in capuchin monkeys (capuchin monkey hepatitis B virus [CMHBV]). We found CMHBV-specific antibodies in five animals and high CMHBV concentrations in one animal. Non-inflammatory, probably chronic infection was consistent with an intact preCore domain, low genetic variability, core deletions in deep sequencing, and no elevated liver enzymes. Cross-reactivity of antisera against surface antigens suggested antigenic relatedness of HBV, CMHBV, and WMHBV. Infection-determining CMHBV surface peptides bound to the human HBV receptor (human sodium taurocholate co-transporting polypeptide), but preferentially interacted with the capuchin monkey receptor homologue. CMHBV and WMHBV pseudotypes infected human hepatoma cells via the human sodium taurocholate co-transporting polypeptide, and were poorly neutralised by HBV vaccine-derived antibodies, suggesting that cross-species infections may be possible. Ancestral state reconstructions and sequence distance comparisons associated HBV with humans, whereas primate hepadnaviruses as a whole were projected to NHP ancestors. Co-phylogenetic analyses yielded evidence for co-speciation of hepadnaviruses and New World NHP. Bayesian hypothesis testing yielded strong support for an association of the HBV stem lineage with hominoid ancestors. Neither CMHBV nor WMHBV was likely the ancestor of the divergent human HBV genotypes F/H found in American natives. CONCLUSIONS: Our data suggest ancestral co-speciation of hepadnaviruses and NHP, and an Old World origin of the divergent HBV genotypes F/H. The identification of a novel primate hepadnavirus offers new perspectives for urgently needed animal models of chronic hepatitis B. LAY SUMMARY: The origins of HBV are unclear. The new orthohepadnavirus species from Brazilian capuchin monkeys resembled HBV in elicited infection patterns and could infect human liver cells using the same receptor as HBV. Evolutionary analyses suggested that primate HBV-related viruses might have emerged in African ancestors of New World monkeys millions of years ago. HBV was associated with hominoid primates, including humans and apes, suggesting evolutionary origins of HBV before the formation of modern humans. HBV genotypes found in American natives were divergent from those found in American monkeys, and likely introduced along prehistoric human migration. Our results elucidate the evolutionary origins and dispersal of primate HBV, identify a new orthohepadnavirus reservoir, and enable new perspectives for animal models of hepatitis B.

Limited Evidence for Infection of Urban and Peri-urban Nonhuman Primates with Zika and Chikungunya Viruses in Brazil.

Chikungunya virus (CHIKV) and Zika virus (ZIKV) emerged in the Americas in 2013. Limited antigenic variability of CHIKV and ZIKV may restrict urban transmission cycles due to population protective immunity. In Africa, sylvatic transmission cycles involving nonhuman primates (NHP) are known for CHIKV and ZIKV, causing cyclic reemergence in humans. To evaluate whether sylvatic cycles can be expected in Latin America, we tested 207 NHP collected between 2012 and 2017 in urban and peri-urban settings in Brazil for infection with ZIKV and CHIKV. No animal tested positive for viral RNA in genus-specific and species-specific reverse transcription-PCR (RT-PCR) assays. In contrast, six animals (2.9%) from the families Atelidae, Callitrichidae, and Cebidae showed ZIKV-specific antibodies and 11 (5.3%) showed CHIKV-specific antibodies in plaque reduction neutralization tests (PRNT). Reactivity was monotypic against either ZIKV or CHIKV in all cases, opposing unspecific virucidal activity of sera. PRNT endpoint titers were low at 1:40 in all NHP, and positive specimens did not correspond to the likely dispersal route and time of introduction of both arboviruses. All antibody-positive samples were therefore tested against the NHP-associated yellow fever virus (YFV) and Mayaro virus (MAYV) and against the human-associated dengue virus (DENV) by PRNT. Two ZIKV-positive samples were simultaneously DENV positive and two CHIKV-positive samples were simultaneously MAYV positive, at titers of 1:40 to 1:160. This suggested cross-reactive antibodies against heterologous alphaviruses and flaviviruses in 24% of ZIKV-positive/CHIKV-positive sera. In sum, low seroprevalence, invariably low antibody titers, and the distribution of positive specimens call into question the capability of ZIKV and CHIKV to infect New World NHP and establish sylvatic transmission cycles. IMPORTANCE Since 2013, Zika virus (ZIKV) and chikungunya virus (CHIKV) have infected millions of people in the Americas via urban transmission cycles. Nonhuman primates (NHP) are involved in sylvatic transmission cycles maintaining ZIKV and CHIKV in the Old World. We tested NHP sampled during 2012 to 2017 in urban and peri-urban areas severely affected by ZIKV and CHIKV in Brazil. Seroprevalence and antibody titers were low for both viruses. Additionally, we found evidence for infection by heterologous viruses eliciting cross-reactive antibodies. Our data suggest that urban or peri-urban NHP are not easily infected by ZIKV and CHIKV despite intense local transmission. These data may imply that the ZIKV and CHIKV outbreaks in the Americas cannot be sustained in urban or peri-urban NHP once human population immunity limits urban transmission cycles. Investigation of diverse animals is urgently required to determine the fate of the ZIKV and CHIKV outbreaks in the Americas.

Formação de educadores para promoção da saúde única em distrito sanitário de Salvador: estudo piloto / Training of educators for one health promotion in a sanitary district of Salvador: pilot study / Formación de educadores para la promoción de la salud única en distrito sanitario de Salvador: estudio piloto

No Brasil, um país de clima tropical e grande diversidade sociocultural, as doenças infectocontagiosas representam um desafio, e a educação em saúde pode ter um poder significativo no combate às doenças zoonóticas. O objetivo deste artigo é identificar e classificar informações sobre agravos zoonóticos, para aplicá-las sob a forma de curso para professores da rede pública. Foi realizada uma pesquisa-intervenção no Distrito Sanitário Cabula/Beirú, em Salvador, Bahia, Brasil. Paralelamente, foram avaliados conhecimentos desses professores sobre temas relacionados à Saúde Única para direcionar a prática educativa para a prevenção de zoonoses e promoção da saúde humana e animal. Os dados obtidos no Sistema Nacional de Agravos de Notificação subsidiaram os questionários aplicados a 32 educadores e 324 educandos do ensino fundamental para a construção do Curso de Formação de Multiplicadores, intervenção educativa online, realizada por meio da Plataforma Moodle da Universidade Federal da Bahia. Foram realizados pré e pós-testes, avaliados pelo Método Soma. Em ambos os grupos, educadores e educandos, os pré-testes evidenciaram desconhecimento, tanto sobre a ocorrência das doenças quanto sobre as medidas preventivas. Esses dados nortearam a prática educativa, que resultou em 100% de eficiência na aprendizagem da maioria dos tópicos e multiplicação nas classes, conforme resultados evidenciados no pós-teste. O curso despertou interesse na comunidade escolar, com 63,3% de preenchimento das vagas por professores e 23,3% por outros interessados, restando 13,4%. Entretanto o índice de evasão foi de 60%. Concluiu-se que a educação online é efetiva em formar educadores multiplicadores, porém o incentivo para a participação dos profissionais requer diálogo entre gestores das áreas de saúde e educação.

Brazil is a country with tropical climate and high socio-cultural diversity, where infectious diseases are a challenge and health education can be significant against zoonotic diseases. The purpose of this study is to identify and classify zoonotic disease information in order to apply it as a course for public school educators. An intervention research was conducted in Cabula/Beirú Sanitary District in Salvador, Bahia, Brasil. At the same time, teachers' knowledge about issues related to the One Health concept was evaluated in order to guide the educational practice for the prevention of zoonoses and the promotion of human and animal health. Data obtained from Notifiable Diseases Information System were subsidy for the questionnaires applied to 32 educators and 324 primary school students for the construction of the Multiplier Training Course, an online educational intervention carried out by means of the Federal University of Bahia Moodle Platform. Pre and post-tests were evaluated, using the Soma Method. In both groups, educators and students, the pre-tests evidenced ignorance, both about the occurrence of diseases and preventive measures. These data guided the educational practice, which resulted in 100% efficiency on learning the majority of the topics and multiplication in classes, according to the results evidenced in the post-test. The course attracted the interest of school community, with 63.3% of attendances by professors and 23.3% by other interested people, remaining 13.4%. However, the evasion rate was 60%. Online education is effective in educating multipliers but, encouraging the participation of professionals requires dialogue between health and education managers.

En Brasil, un país de clima tropical y de gran diversidad sociocultural, las enfermedades infecciosas representan un reto; y la educación en salud puede tener un poder significativo en el combate a las enfermedades zoonóticas. Con el objetivo de identificar y clasificar información sobre los agravios zoonóticos, para poder aplicarla en forma de cursos para maestros de la red pública. Fue realizada una investigación-intervención en el Distrito Sanitario Cabula/Beirú de Salvador, Bahia, Brasil. Paralelamente, fueron evaluados los conocimientos de estos maestros sobre los temas relacionados a la Salud Única para direccionar la práctica educativa hacia la prevención de zoonosis y la promoción de la salud humana y animal. Datos obtenidos en el Sistema de Información de Agravios de Notificación ofrecieron subsidio a los cuestionarios aplicados a 32 educadores y 324 educandos de la educación básica para la construcción del Curso de Formación de Multiplicadores, intervención educativa online, realizada a través de la Plataforma Moodle de la Universidad Federal de Bahia. Fueron realizadas pruebas preliminares y pruebas posteriores, evaluadas por el Método Soma. En ambos grupos, educadores y educandos, las pruebas preliminares evidenciaron desconocimiento, tanto sobre la ocurrencia de las enfermedades, como sobre las medidas preventivas. Estos datos sirvieron como referencia para la práctica educativa, que resultó en 100% de eficacia en el aprendizaje de la mayoría de los tópicos y multiplicación en las clases, conforme los resultados evidenciados en las pruebas posteriores. El curso despertó interés en la comunidad escolar, rellenóse el 63,3% de las plazas por maestros y el 23,3% por otros interesados, restando 13,4%. Sin embargo, el índice de evasión fue de 60%. La educación online es efectiva para formar a los educadores multiplicadores, todavía, el incentivo para la participación de los profesionales requiere diálogo entre los gestores de las áreas de salud y educación.

Toxoplasmatinae Parasites in Bats from Bahia State, Brazil.

Toxoplasma gondii and Neospora caninum are widespread cyst-forming coccidian parasites of the subfamily Toxoplasmatinae that infect a wide range of wild and domestic animals. Whereas T. gondii is a zoonotic disease, N. caninum is restricted to nonhuman animals. Some chiropteran species can be infected by T. gondii and present fatal toxoplasmosis. In most cases, T. gondii -infected bats are believed to remain asymptomatic and to act as an infection source to other animals. It is not known whether N. caninum can infect bats. We determined infection rates of T. gondii and N. caninum in free-living bats in the state of Bahia, Brazil. Brain samples from 97 bats of seven species, captured in 2008-15, were analyzed by PCRs for T. gondii and N. caninum . Two of the 97 samples were positive for T. gondii DNA. None of the samples were positive for N. caninum DNA, suggesting that the bats were not susceptible to N. caninum infection or that its prevalence was very low.

Xenodiagnosis on dogs with visceral leishmaniasis: Canine and sand fly aspects related to the parasite transmission.

One of the main limitations for the effective control of canine leishmaniasis in endemic areas is the difficulty in identifying infectious dogs. The objective of this study was to determine factors, related to dogs and to parasite detection in sand flies, which are associated with the positive xenodiagnosis of Leishmania infantum using the vector Lutzomyia longipalpis. The xenodiagnosis was performed in 50 owned dogs residing in endemic areas, which were divided into three different groups: G1-26 dogs proved to be infected and classified by severity of VL clinical signs on physical examination; G2-15 dogs proved to be infected and classified by severity of clinical signs and intensity of laboratory abnormalities; G3-nine dogs that were seropositive for anti-Leishmania IgG in ELISA tests. Parasite search in the sand flies after having fed on dogs in the xenodiagnosis was performed by both methodologies, PCR and dissection followed by microscopy. In G1, 58% (15/26) of dogs were able to transmit Leishmania to the vector, when parasite detection in sand flies were performed by PCR technique, 5 days after blood meal, whereas in G2, 53% (8/15) transmitted the parasite to the vector, however, confirmation was performed by direct observation of parasite through optical miscroscopy held 10 days after blood meal. Rate of infectiousness of dogs to sand flies was positively associated to severity of disease (p=0.042 and p=0.040), regardless the method used for clinical classification or for parasite detection in sand flies after xenodiagnosis. In G1 30% (3/10) of dogs with subclinical infection were infectious to the vector, while 80% (12/16) of dogs with clinical disease were also infectious. Even more, 17% (1/6) of dogs that had moderate disease were infectious to the sand flies, while 78% (7/9) of dogs with severe disease were infectious in G2. Still in G2, the proportion of sand flies infected (grade of infectiousness) was significantly lower (p=0.0098) when they fed on dogs with moderate disease (1%) in comparison with dogs with severe disease (38%). The dogs from G3 presented a rate of infectiousness of 11% (1/9), demonstrating that the indirect ELISA is not a good indicator of infectiousness and, therefore, should not be used as a confirmatory test for the euthanasia of dogs, as it is currently done in Brazil.

Trace elements distribution in hawksbill turtle (Eretmochelys imbricata) and green turtle (Chelonia mydas) tissues on the northern coast of Bahia, Brazil.

Concentrations of elements (As, Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Sb, Se, Sr, V, Zn) were determined in liver, kidneys and bones of Eretmochelys imbricata and Chelonia mydas specimens found stranded along the northern coast of Bahia, Brazil. Results showed that the concentrations of Cd, Cu, Ni and Zn in the liver and kidneys of juvenile C. mydas were the highest found in Brazil. We also observed a significant difference (p<0.05) on the bioaccumulation of trace elements between the two species: Al, Co, Mo, Na and Se in the liver; Al, Cr, Cu, K, Mo, Ni, Pb, Sr and V in the kidneys; and Al, Ba, Ca, Cd, Mn, Ni, Pb, Se, Sr and V in the bones. This study represents the first report on the distribution and concentration of trace elements in E. imbricata in the Brazilian coast.

Identification of biomarkers in the hair of dogs: new diagnostic possibilities in the study and control of visceral leishmaniasis.

Visceral leishmaniasis (VL) is a zoonosis whose etiologic agent in the Americas is Leishmania infantum, and dogs are the main host. Research and innovation in diagnostic techniques are essential to improve the surveillance and control of VL in endemic areas. The present study investigates the profile of the volatile organic compounds (VOCs) emitted by healthy dogs and by dogs infected by L. infantum to detect variations in the VOCs that may be used as biomarkers in the diagnosis of VL. In total, 36 dogs were selected from an endemic area and divided into three groups: G1, not infected with L. infantum; G2, infected without clinical signs of VL; and G3, infected with clinical signs of VL. To analyze the profiles of the VOCs emitted by dogs from the three groups, solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) was used. Variations were observed between the profiles of the VOCs emitted in the three groups studied, and they also differentiated infected animals with or without clinical signs. Six VOCs were identified as potential biomarkers of infection, with significant variations between healthy dogs (G1) and infected dogs (G2 + G3). The detection of variations between groups G2 and G3 suggested that the profiles of some VOCs may be related to the type of immune response and the parasite load of the infected dogs. This study demonstrated the possibility of analysis of VOCs as biomarkers of VL in diagnostic, clinical, and epidemiological work.

Bats carry pathogenic hepadnaviruses antigenically related to hepatitis B virus and capable of infecting human hepatocytes.

The hepatitis B virus (HBV), family Hepadnaviridae, is one of most relevant human pathogens. HBV origins are enigmatic, and no zoonotic reservoirs are known. Here, we screened 3,080 specimens from 54 bat species representing 11 bat families for hepadnaviral DNA. Ten specimens (0.3%) from Panama and Gabon yielded unique hepadnaviruses in coancestral relation to HBV. Full genome sequencing allowed classification as three putative orthohepadnavirus species based on genome lengths (3,149-3,377 nt), presence of middle HBV surface and X-protein genes, and sequence distance criteria. Hepatic tropism in bats was shown by quantitative PCR and in situ hybridization. Infected livers showed histopathologic changes compatible with hepatitis. Human hepatocytes transfected with all three bat viruses cross-reacted with sera against the HBV core protein, concordant with the phylogenetic relatedness of these hepadnaviruses and HBV. One virus from Uroderma bilobatum, the tent-making bat, cross-reacted with monoclonal antibodies against the HBV antigenicity determining S domain. Up to 18.4% of bat sera contained antibodies against bat hepadnaviruses. Infectious clones were generated to study all three viruses in detail. Hepatitis D virus particles pseudotyped with surface proteins of U. bilobatum HBV, but neither of the other two viruses could infect primary human and Tupaia belangeri hepatocytes. Hepatocyte infection occurred through the human HBV receptor sodium taurocholate cotransporting polypeptide but could not be neutralized by sera from vaccinated humans. Antihepadnaviral treatment using an approved reverse transcriptase inhibitor blocked replication of all bat hepadnaviruses. Our data suggest that bats may have been ancestral sources of primate hepadnaviruses. The observed zoonotic potential might affect concepts aimed at eradicating HBV.

Highly diversified coronaviruses in neotropical bats.

Bats host a broad diversity of coronaviruses (CoVs), including close relatives of human pathogens. There is only limited data on neotropical bat CoVs. We analysed faecal, blood and intestine specimens from 1562 bats sampled in Costa Rica, Panama, Ecuador and Brazil for CoVs by broad-range PCR. CoV RNA was detected in 50 bats representing nine different species, both frugivorous and insectivorous. These bat CoVs were unrelated to known human or animal pathogens, indicating an absence of recent zoonotic spill-over events. Based on RNA-dependent RNA polymerase (RdRp)-based grouping units (RGUs) as a surrogate for CoV species identification, the 50 viruses represented five different alphacoronavirus RGUs and two betacoronavirus RGUs. Closely related alphacoronaviruses were detected in Carollia perspicillata and C. brevicauda across a geographical distance exceeding 5600 km. Our study expands the knowledge on CoV diversity in neotropical bats and emphasizes the association of distinct CoVs and bat host genera.

Positividade para leishmaniose visceral canina: existem fatores caninos que contribuem? / Positividad para la leishmaniasis visceral canina: ¿existen factores caninos que contribuyen? / Positivity for canine visceral leishmaniasis: are there canine factors contributing?

A leishmaniose visceral canina (LVC), doença endêmica no Litoral Norte do Estado da Bahia, vem sendo investigada em seus aspectos epidemiológicos, ambientais e sociais, buscando-se formas de minimizar a sua expansão. O presente estudo de caso-controle objetivou identificar características inerentes aos cães de área endêmica e correlacioná-las àpositividade para LVC. Foram avaliadas as fichas clínicas de 226 cães testados sorologicamente no Laboratório de Infectologia Veterinária da Universidade Federal da Bahia, por meio do teste ELISA para soropositividade para anticorpos antileishmania, nos anos de 2009 e 2010,sendo selecionados todos os casos positivos nesses dois anos, totalizando 23; foram escolhidos 27 controles negativos com base nas fichas dos animais testados sob as mesmas condições. A soropositividade foi correlacionada às variáveis sexo, idade, raça e procedência. Para a análise estatística, foi escolhido o Teste Exato de Fisher, realizado pelo software Stata versão 10, e calculados o p-valor, a odds ratio e o intervalo de confiança. Foi possível encontrar umarelação incipiente entre sexo masculino e soropositividade, bem como uma forte relação entre procedência e soropositividade para LVC. São necessários estudos com um maior número de cães, buscando-se relacionar estas características à positividade para LVC, como forma deconhecer características caninas que propiciem a infecção.

The Canine Visceral Leishmaniasis (CVL) an endemic disease in the northern coast of Bahia has been investigated in its epidemiological, environmental and social aspects. The investigation seeks to find ways to minimize its spread. This study aimed to evaluate some characteristics of dogs and to correlate them to the positivity for CVL, using a casecontrolstudy. The medical records of 226 dogs tested serologically through the use of ELISA kit for CVL were evaluated at the Laboratory of Veterinary Infectious Diseases at the Federal University of Bahia, during the years 2009 and 2010.The total of positive cases were 23. There were chosen 27 dogs that had negative results based on forms of animals tested in the same conditions. Seropositivity was associated with gender, age, breed and origin. The Fisher's Exact Test was performed for statistical analysis, using the software Stata version 10.The p-value was calculated and the odds ratio and confidence intervals as well. It was possible to find evidence of association between the male sex and positivity as well as a strong association between origin and seropositivity for CVL. Larger studies with a greater number of dogs are needed to investigate the characteristics of positive animals for CVL as a way of knowing canine factors that are conducive to disease.

La leishmaniasis visceral canina (LVC), una enfermedad endémica en lacosta norte de la Bahía, ha sido investigada por sus aspectos epidemiológicos, ambientales y sociales, buscando formas que minimizen su propagación. Este estudio de caso-control tuvo como objetivo identificar características inherentes a los perros del área endémica ycorrelacionarlas con la positividad para LVC. Se evaluaron las historias clínicas de 226 perros testados serológicamente en el Laboratorio de Infectología Veterinaria de la Universidad Federal de la Bahía, por medio del test ELISA para seropositividad para anticuerpos antileishmania, en los años 2009 y 2010, siendo seleccionados todos los casos positivos en esos dos años, totalizando 23, fueron seleccionados 27 controles negativos basados en los registros de los animales testados bajo las mismas condiciones. La seropositividad fue correlacionada con las variables sexo, edad, raza y origen. Para el análisis estadísticofue elegido el Test Exacto de Fisher, realizado con el software Stata versión 10, y calculado el p-valor, la oddsratio, e el intervalo de confianza. Fue posible encontrar una incipiente relación entre el sexo masculino y seropositividad, así como, una fuerte relación entre origen y seropositividad para LVC. Se necesitan estudios con un mayor número de perros, buscando relacionar estas características con la positividad para LVC, como una manera de conocer las características caninas que propician la enfermedad