The Amazonian Tropical Bites Research Initiative, a hope for resolving zoonotic neglected tropical diseases in the One Health era.

BACKGROUND: Neglected tropical diseases (NTDs) disproportionately affect populations living in resource-limited settings. In the Amazon basin, substantial numbers of NTDs are zoonotic, transmitted by vertebrate (dogs, bats, snakes) and invertebrate species (sand flies and triatomine insects). However, no dedicated consortia exist to find commonalities in the risk factors for or mitigations against bite-associated NTDs such as rabies, snake envenoming, Chagas disease and leishmaniasis in the region. The rapid expansion of COVID-19 has further reduced resources for NTDs, exacerbated health inequality and reiterated the need to raise awareness of NTDs related to bites. METHODS: The nine countries that make up the Amazon basin have been considered (Bolivia, Brazil, Colombia, Ecuador, French Guiana, Guyana, Peru, Surinam and Venezuela) in the formation of a new network. RESULTS: The Amazonian Tropical Bites Research Initiative (ATBRI) has been created, with the aim of creating transdisciplinary solutions to the problem of animal bites leading to disease in Amazonian communities. The ATBRI seeks to unify the currently disjointed approach to the control of bite-related neglected zoonoses across Latin America. CONCLUSIONS: The coordination of different sectors and inclusion of all stakeholders will advance this field and generate evidence for policy-making, promoting governance and linkage across a One Health arena.

Assessment of crab fishermen's exposure to rabies virus in a typical Amazonian community.

Outbreaks of human rabies transmitted by hematophagous bats occurred in 2018 in Pará state, Brazil, eastern Amazon, after 12 years of no record of the disease. Thus, it is necessary to understand the epidemiological characteristics of these attacks to protect the local population. This study aimed to characterize the bat bite populations in the municipality of São João da Ponta, Pará State, Brazil, in 2013-2015. The Notifiable Diseases Information System (SINAN) database was used to identify the five individuals who sought medical care during the study period (seeds). Other individuals who were attacked during the same period but did not seek medical care (n = 61) were reached by snowball sampling, and a descriptive analysis was performed based on information obtained from questionnaires. Majority of the interviewees were men (92.4%; 61/66) and adults aged 20-50 years (69.9%; 46/66) and had <4 years of formal school education (86.3%; 57/66). Additionally, most of them were rural residents (92.4%; 61/66) and crab fishermen (79.3%; 53/66). The interviewees (92.4%; 61/66) identified mangroves of the Mãe Grande de Curuçá extractive reserve, where groups of fishermen sometimes gather for several days for crab fishing, often living in improvised dwellings without walls and covered by tarps or straw (88.8%; 56/66), conducive to attacks by vampire bats. Overall, 42.4% (28/66) of the participants had been bitten more than four times throughout their life. The median number of attacks over the participant's lifetime was 3.11 (range, 1-23). Participants were unaware of the risk of contracting rabies from the bite (95.4%; 65/66). These results suggest that vampire bat attacks are essentially an occupational hazard in the study region. Moreover, for each reported attack, there were at least 12.2 unreported cases. Thus, the study highlights the need to develop strategies for prophylactic treatment of this population.

Serological prevalence of Toxoplasma gondii infection in cats (Belém, Pará, Brazil).

We evaluated the prevalence of anti-Toxoplasma gondii antibodies in the serum samples collected from domestic cats in Belém, Pará, Brazil. We also correlated the presence of T. gondii antibodies with environmental variables and cat-owner habits. Four-hundred and forty-seven serum samples from domestic cats were analyzed. The sera were tested using an indirect immunofluorescence assay. Among the animals analyzed, 21.92% (98/447) were seropositive. A statistically significant association was found in relation to age and serology among the animals over 1 year old (p<0.01): in the group up to 1 year old, 12.82% (20/156) of the animals were positive, and in the group over 1 year old, 26.80% (78/291) were positive. Our results show that the cats in Belém, Pará region have anti-T. gondii antibodies, and their owners are not aware of toxoplasmosis or how to prevent its transmission.

Livestock rabies in Pará state, Brazil: a descriptive study (2004 to 2013) / Raiva em herbívoros no estado do Pará, Brasil: estudo descritivo (2004 a 2013)

Rabies is an important zoonosis to public health associated with lethal encephalitis and economic losses. Analysis of its spatial distribution is a meaningful tool in understanding its dispersion, which may contribute to the control and prophylaxis of the disease. This study analyzed the spatial-temporal distribution of rabies outbreaks in livestock in Pará state, Brazil, from 2004 to 2013. We used records of neurological syndromes obtained from the state's livestock authority (Adepará). The analysis recorded 711 neurological syndromes reports in livestock, of which 32.8% were positive for rabies. In 8% of the neurological syndromes (n=57) was not possible to perform the analysis because of bad-packaging conditions of the samples sent. Outbreaks involved at least 1,179 animals and cattle were the most affected animal species (76.8%). The numbers of reported neurological syndromes and of rabies outbreak shad strong positive correlation and exhibited decreasing linear trend. Spatially, most outbreaks occurred in two mesoregions in Pará (Northeast and Southeast). One of the justifications for this spatial distribution may be related with the distribution of the animals in the state, since these mesoregions are the largest cattle producers in Pará and have most of their territory deforested for pasture implementation.(AU)

A raiva é uma zoonose importante para a saúde pública associada à encefalite letal e às perdas econômicas. A análise de sua distribuição espacial é uma ferramenta importante no entendimento de sua dispersão, o que pode contribuir para o controle e a profilaxia da doença. Este estudo analisou a distribuição espaço-temporal do surto de raiva em rebanhos no estado do Pará, Brasil, entre 2004 e 2013. Foram utilizados registros de síndromes neurológicas obtidas junto à agência de defesa agropecuária do estado (Adepará). A análise revelou 711 notificações de síndromes neurológicas em herbívoros, das quais 32,8% foram positivas para raiva. Em 8% das síndromes neurológicas (n=57) não foi possível realizar as análises devido às más condições das amostras enviadas. Surtos envolveram pelo menos 1.179 animais e os bovinos foram a espécie animal mais afetada (76,8%). Os números de síndromes neurológicas relatadas e de surtos de raiva apresentam forte correlação positiva e exibem tendência linear decrescente. Espacialmente, a maioria dos surtos ocorreu em duas mesorregiões no Pará (Nordeste e Sudeste). Uma das justificativas para essa distribuição espacial pode estar relacionada à distribuição dos animais no estado, uma vez que essas mesorregiões são os maiores produtores de gado do Pará e possuem grande parte do seu território desflorestado para implantação de pastagens.(AU)

Epidemiological scenarios for human rabies exposure notified in Colombia during ten years: A challenge to implement surveillance actions with a differential approach on vulnerable populations.

Based on notified cases of human rabies exposure and human deaths by rabies to Colombia public health surveillance system between 2007 and 2016, we conducted a spatiotemporal analysis to identify epidemiological scenarios of high human rabies exposure due to dogs, cats, bats, or farm animals (n = 666,411 cases). The incidence rate of human rabies exposures was analyzed by using geographical information system (spatiotemporal distribution and Cluster and Outlier Analysis (Anselin Local Moran's I)) data for all Colombian cities. The incidence rate of human rabies exposures due to dogs and cats showed an increasing trend, while aggression due bats and farm animals fluctuated throughout the analyzed period. Human deaths by rabies transmitted by cat and bat occurred in the Andean and Orinoquia regions, which had urban and rural scenarios. The urban scenario showed the highest exposure to human rabies due to cats and dogs in cities characterized with high human population density and greater economic development. In contrary, the highest human rabies exposure in the rural scenario was observed due to contact of mucosa or injured skin with the infected saliva of farm animals with the rabies virus, principally among workers in the agroforestry area. The inequality scenario showed some outlier cities with high human rabies exposure due to farm animals principally in the Pacific region (characterized by the highest poverty rates in Colombia), being Afro-descendant and indigenous population the most exposed. The highest exposure due to bats bite was observed among indigenous people residing in cities of the Amazon region as a dispersed population (Amazonian scenario). None of the high exposure scenarios were related to human deaths by rabies due to dogs aggression. The identified scenarios can help develop better surveillance systems with a differential approach to the vulnerable population and strengthening them in areas with rabies viral circulation.

Dirofilaria immitis infection in dogs in Algodoal Island, Brazilian Amazon / Infecção por Dirofilaria immitis em cães na Ilha de Algodoal, Amazônia brasileira

Dirofilaria immitis, a parasite that mainly infects domestic or wild canids, but can infect felines or humans as well, is frequent in many Brazilian areas. The main objective of this research was to determine the prevalence of natural canine infection at the Algodoal-Maiandeua Island complex, in the coastal region of the state of Pará, Brazil. A total of 67 dogs were sampled for blood microfilariae detection and for D. immitis DNA detection. Microfilaria and D. immitis DNA could be detected in 35.8% (24/67) of the animals. In one dog's sample no microfilariae were detected, but the PCR was positive, suggesting that either larvae recently were eliminated or adults died shortly before sample collecting. Therefore, it can be concluded that the occurrence of D. immitis is a health threat for domestic and wild canids at the Island of Algodoal, as well as for feline or human health.(AU)

Dirofilaria immitis, um parasito que infecta principalmente canídeos domésticos ou selvagens, embora também possa infectar felinos e humanos, é frequente em muitas áreas do Brasil. O objetivo deste estudo foi determinar a ocorrência da infecção natural em cães provenientes do complexo da Ilha de Algodoal-Maiandeua, região litorânea do estado do Pará, Brasil. Um total de 67 cães tiveram o sangue coletado para detecção de microfilárias de D. immitis e seu DNA. Microfilárias e o DNA de D. immitis foram detectados em 35,8% (24/67) dos animais. Na amostra de um animal, não foram observadas microfilárias, mas o seu DNA foi detectado, sugerindo que as larvas tenham sido recentemente eliminadas ou os adultos tenham morrido antes da coleta da amostra. Portanto, pode-se concluir que a ocorrência de D. immitis é uma ameaça à saúde de canídeos domésticos no complexo da Ilha de Algodoal-Maiandeua, bem como para felinos e seres humanos.(AU)

Detection of Toxoplasma gondii in Crassostrea spp. oysters cultured in an estuarine region in eastern Amazon.

The aim of the present study was to detect DNA of Toxoplasma gondii in Crassostrea spp. oysters cultured in the state of Pará, Brazil. A total of 400 oysters were directly collected from a fixed rack system. Gills, gastrointestinal tract (GIT) and intervalvular liquid were separated and grouped into pool samples of 10 animals, resulting in 40 samples each of gills, GIT and intervalvular liquid. DNA extraction was performed using a commercial kit, and T. gondii DNA was detected by nested PCR using the primers Toxo3 and Toxo4, which produced an amplification product of 155 bp of the T. gondii gene B1. Nucleotide sequencing was performed for positive samples, and the obtained sequences were identified by comparison with sequences in GenBank. The DNA of T. gondii was detected in 5.8% (7/120) of the pool samples, of which 7.5% (3/40) was in the GIT, 5% (2/40) in the gills, and 5% (2/40) was in the intervalvular liquid. The obtained sequences presented 100% identity and overlap with T. gondii DNA sequences. This is the report of detection of T. gondii DNA in oysters from genus Crassostrea spp. originating from the state of Pará, eastern Amazon.

Avaliação higiênico-sanitária de estabelecimentos comerciais e análise de micro-organismos indicadores em amostras de carne bovina (coxão mole) in natura comercializadas em mercados públicos / Sanitary inspection of commercial establishments and analysisof the microorganism indicators in the bovine pad topside meatsamples sold in public markets

O objetivo deste estudo foi analisar as condições higinicos-sanitárias dos mercados públicos dos municípios da microrregião de Cametá, estado do Pará, e avaliar as características microbiológicas das carnes comercializadas nestes locais por meio da pesquisa de micro-organismos indicadores. Sessenta e quatro amostras de carne bovina in natura foram coletadas e analisadas por meio da determinação do Número Mais Provável de coliformes a 35 oC e 45 oC e contagem de Staphylococcus coagulase positiva, de acordo com a Instrução Normativa no 62 de 2003 do Ministério da Agricultura Pecuária e Abastecimento. Paralelamente, as condições higiênico-sanitárias foram avaliadas aplicando-se checklist. Os resultados obtidos evidenciaram que todos os locais avaliados apresentaram condições higiênico-sanitárias insatisfatórias, sendo classificados como deficientes. As amostras apresentaram Número Mais Provável de coliformes a 35ºC e 45ºC que variaram de 64 NMP/g a >1100NMP/g, e contagens de Staphylococcus coagulase positiva entre 1,8 x 10² UFC/g e 2,3 x 105 UFC/g.Os estabelecimentos avaliados não atenderam às exigências mínimas de higiene e sanidade, e a carnecomercializada apresentou elevada contagem de micro-organismos indicadores.

The aim of this study was to analyze the sanitary conditions of public markets located in theCametá micro-region, state of Pará, and to assess the microbiological condition of the meat sold inthese locations through the determination of microorganism indicators. For this purpose, 64 fresh beefsamples were collected and the determination of the Most Probable Number of coliforms at 35 oC and45 oC and coagulase positive Staphylococcus counts were performed, according to the Instruction no 62,2003 of the Ministry of Agriculture Livestock and Supply. In addition, the hygienic and sanitary conditionsof the public markets were evaluated by applying a checklist. The found results showed that all of theevaluated public markets indicated poor hygienic sanitary condition, and they were ranked as deficient.The samples showed the Most Probable Number of coliforms at 35 oC and 45 oC ranging from 64 MPN/gto >1100NMP/g, and coagulase positive Staphylococcus counts between 1.8 x 10² CFU/g and2.3 x 105 CFU/g. The evaluated food markets did not meet the minimum requirements of hygiene andsanitation and the sold meat samples showed high count of microorganisms indicators.

Artificial feeding of Amblyomma cajennense (Acari: Ixodidae) fasting females through capillary tube technique / Alimentação artificial de fêmeas de Amblyomma cajennense (Acari: Ixodidae) em jejum por meio da técnica de tubos capilares

The present study aimed to adjust the artificial feeding technique through capillaries and to verify its influence over the biology of Amblyomma cajennense females. Five groups of 20 female ticks were formed. Females were starved for 45 days and then fed with citrated bovine blood using capillary tubes in different periods of time. Females were divided in five experimental groups with 20 individuals each and fed as follows: groups uninterruptedly fed for 12, 24, and 48 hours and groups fed 2 and 6 h a day, for a period of 8 days. Subsequently, ticks were exposed to rabbits for complementary feeding and their biological parameters were analyzed. TIcks were capable of feeding, showing rounded idiosoma, visible even to naked eyes, following the feeding period. The groups fed for 24 hours, 2 hours/day for eight consecutive days or 6h/day for eight consecutive days presented greater weight gain, without statistically significant differences. These results suggested that 24 hours of artificial feeding were enough for fasting females to increase weight by 2.43 mg. Artificial feeding through capillaries did not interfere with parasitic and non-parasitic phases of A. cajennense females.

O presente trabalho teve como objetivos adaptar a técnica de alimentação artificial por meio de tubos capilares para fêmeas de Amblyomma cajennense e verificar a sua influência sobre os parâmetros biológicos da espécie. Fêmeas em jejum por 45 dias foram alimentadas com sangue bovino citratado usando tubos capilares por diferentes períodos de tempo: 12, 24 e 48 horas, 2 horas/dia e 6 horas/dia por 8 dias consecutivos, sendo que cada grupo foi composto por 20 carrapatos. Em seguida, os carrapatos foram expostos a coelhos para completar a alimentação e seus parâmetros biológicos foram analisados. Após alimentação artificial, os carrapatos apresentaram idiossoma arredondado mesmo à vista desarmada. Os grupos 24 horas, 8 dias/2 horas/dia e 8 dias/6 horas/dia tiveram maior ganho de peso, não havendo diferença estatística entre eles. Desta forma, nas condições do presente trabalho, 24 horas de alimentação artificial foram suficientes para que fêmeas em jejum aumentassem seu peso em 2,43mg. A alimentação artificial utilizando tubos capilares não interferiu nos parâmetros relativos às fases parasitária e não-parasitária de fêmeas A. cajennense.

Artificial feeding of Amblyomma cajennense (Acari: Ixodidae) fasting females through capillary tube technique.

The present study aimed to adjust the artificial feeding technique through capillaries and to verify its influence over the biology of Amblyomma cajennense females. Five groups of 20 female ticks were formed. Females were starved for 45 days and then fed with citrated bovine blood using capillary tubes in different periods of time. Females were divided in five experimental groups with 20 individuals each and fed as follows: groups uninterruptedly fed for 12, 24, and 48 hours and groups fed 2 and 6 h a day, for a period of 8 days. Subsequently, ticks were exposed to rabbits for complementary feeding and their biological parameters were analyzed. TIcks were capable of feeding, showing rounded idiosoma, visible even to naked eyes, following the feeding period. The groups fed for 24 hours, 2 hours/day for eight consecutive days or 6h/day for eight consecutive days presented greater weight gain, without statistically significant differences. These results suggested that 24 hours of artificial feeding were enough for fasting females to increase weight by 2.43 mg. Artificial feeding through capillaries did not interfere with parasitic and non-parasitic phases of A. cajennense females.

Borrelia sp. in naturally infected Didelphis aurita (Wied, 1826) (Marsupialia: Didelphidae)

Cinquenta e seis gambás (Didelphis aurita) foram capturados no campus da Universidade Federal Rural do Rio de Janeiro, Seropédica - Rio de Janeiro, com o intuito de se investigar a ocorrência de Borrelia sp. entre eles, associada ao estudo da espiroquetemia e de seus ectoparasitos. Amostras de sangue dos animais foram examinadas à microscopia de campo escuro e contraste de fase, além deste ter se destinado à obtençäo de esfregaços sanguíneos, corados pelo método de Giemsa. Näo houve relaçäo entre os resultados obtidos através dos exames diretos do sangue (13 gambás positivos para Borrelia sp.) e aqueles observados nos esfregaços sanguíneos (dois animais positivos), o que demonstra a baixa eficácia desta técnica. Estudos sobre a parasitemia de 37 animais mantidos em cativeiro, assim como as várias recapturas nas quais animais uma vez negativos mostraram-se positivos dias mais tarde, revelaram que os estudos hemoscópicos podem ser utilizados eficientemente como ferramentas de diagnóswtico. Ectoparasitas colhidos de nove animais foram classificados. Houve ocorrência de ninfas Amblyomma cajannense e adultos Ctenocephalides sp.