A rapid inventory of amphibians, squamates, and bats of Mata de Plátano Field Station and Nature Reserve, Arecibo, Puerto Rico.

Puerto Rico harbors a diverse vertebrate fauna with high levels of endemism. However, while several books for vertebrate diversity and local checklists for birds have been published, checklists of amphibians, reptiles, and bats are lacking or nonexistent at both local and regional scales. In this study, we documented the amphibian, reptile, and bat faunas at Mata de Plátano Field Station and Nature Reserve, in Arecibo, Puerto Rico. We document four species of amphibians, seven lizards, three snakes, and nine bats. Despite prior works using samples from this nature reserve, this represents the first vertebrate checklist for the Mata de Plátano Field Station and Nature Reserve. We provide additional natural history observations based on our survey results and highlight the importance of including local and regional checklists of species for downstream research and conservation.

Epidemiological profile of accidents involving venomous animals in Maranhão from 2012 to 2021 / Perfil epidemiológico de acidentes envolvendo animais peçonhentos no Maranhão no período de 2012 a 2021 / Perfil epidemiológico de los accidentes con animales venenosos en Maranhão de 2012 a 2021

Background and Objectives: the second greatest cause of human poisoning in Brazil is caused by venomous animals. Thus, this study aimed to analyze clinical and sociodemographic data, in order to outline the epidemiological profile of accidents involving venomous animals in Maranhão. Methods: this is an ecological study with a quantitative approach, carried out from data collection by the Notifiable Diseases Information System (SINAN) of notifications of accidents by venomous animals that occurred in the state of Maranhão from 2012 to 2021. Results: of the 34,808 cases reported, it was found that the highest incidence occurred in 2019 and, in general, January is the month in which most accidents are recorded. When analyzing the sociodemographic profile, it is noted that the majority of victims are represented by male individuals, between 20-39 years old, with an unidentified level of education. In the case of clinical and epidemiological criteria, it was evident that snakes of the genus Bothrops are responsible for the highest incidence, with most cases involving venomous animals occurring within 1-3 hours after the bite. Among the cases identified, 63% were described as mild and 82% progressed to cure. Conclusion: the epidemiological profile described in the study can be used by health agents to plan preventive measures in Primary Health Care, and knowing the profile of victims is essential to prevent and promote quality care.(AU)

Justificativa e Objetivos: a segunda maior causa de envenenamento humano no Brasil é ocasionada por animais peçonhentos. Dessa forma, este estudo visou analisar dados clínicos e sociodemográficos, a fim de traçar o perfil epidemiológico dos acidentes envolvendo animais peçonhentos no Maranhão. Métodos: trata-se de estudo ecológico de abordagem quantitativa, realizado a partir da coleta de dados pelo Sistema de Informações de Agravos de Notificação (SINAN) de acidentes por animais peçonhentos ocorridos no estado do Maranhão no período de 2012 a 2021. Resultados: dos 34.808 casos notificados, constatou-se que a maior incidência ocorreu no ano de 2019 e, em geral, janeiro é o mês em que mais se registram acidentes. Ao analisar o perfil sociodemográfico, nota-se que a maior parte das vítimas é representada por indivíduos do sexo masculino, entre 20 e 39 anos, com grau de escolaridade não identificado. Tratando-se de critérios clínicos e epidemiológicos, evidenciou-se que as serpentes do gênero Bothrops são as responsáveis pela maior incidência, sendo que a maior parte dos atendimentos envolvendo animais peçonhentos ocorreu no intervalo de 1-3 horas após a picada. Entre os casos identificados, 63% foram descritos como leve e 82% evoluíram para a cura. Conclusão: o perfil epidemiológico descrito no estudo pode ser utilizado por agentes de saúde para o planejamento de medidas preventivas na Atenção Primária à Saúde, e conhecer o perfil das vítimas é essencial para prevenir e promover uma assistência de qualidade.(AU)

Justificación y Objetivos: la segunda causa de envenenamiento humano en Brasil es causada por animales venenosos. Así, este estudio tuvo como objetivo analizar datos clínicos y sociodemográficos, con el fin de delinear el perfil epidemiológico de los accidentes con animales venenosos en Maranhão. Métodos: se trata de un estudio ecológico con abordaje cuantitativo, realizado a partir de la recolección de datos por el Sistema de Información de Enfermedades de Declaración Obligatoria (SINAN) de relatos de accidentes por animales ponzoñosos ocurridos en el estado de Maranhão entre 2012 y 2021. Resultados: de los 34.808 casos reportados, se encontró que la mayor incidencia ocurrió en 2019 y, en general, enero es el mes en el que se registran más accidentes. Al analizar el perfil sociodemográfico, se observa que la mayoría de las víctimas están representadas por personas del sexo masculino, entre 20-39 años, y con nivel de escolaridad no identificado. En el caso de los criterios clínicos y epidemiológicos, se evidenció que las serpientes del género Bothrops son las responsables de la mayor incidencia, y la mayoría de los casos que involucran animales venenosos ocurren entre 1 y 3 horas después de la mordedura. Entre los casos identificados, el 63% fueron descritos como leves y el 82% progresó hacia la curación. Conclusión: el perfil epidemiológico descrito en el estudio puede ser utilizado por los agentes de salud para planificar medidas preventivas en la atención básica de salud, y conocer el perfil de las víctimas es fundamental para prevenir y promover una atención de calidad.(AU)

Comparative morphology of oral glands in snakes of the family Homalopsidae reveals substantial variation and additional independent origins of salt glands within Serpentes.

Using diffusible iodine-based contrast-enhanced computed tomography (diceCT), we examined the morphology of the oral glands of 12 species of the family Homalopsidae. Snakes of this family exhibit substantial interspecific morphological variation in their oral glands. Particular variables are the venom glands, ranging from large (e.g., Subsessor bocourti) to small (e.g., Erpeton tentaculatum). The supra- and infralabial glands are more uniform in morphology, being the second most developed in almost all the sampled species. Premaxillary glands distinct from the supralabial glands were observed in five species (Myron richardsonii, Bitia hydroides, Cantoria violacea, Fordonia leucobalia, and Gerarda prevostiana), in addition to Cerberus rynchops, the only species in which this condition was previously documented associated with the excretion of salt. In the three species of the saltwater group of homalopsids (C. violacea, F. leucobalia, and G. prevostiana), the premaxillary glands also extend posteriorly, occupying a large area above the supralabial gland, a condition not observed in any other species of snake studied thus far. Character evolution analyses indicate that premaxillary glands differentiated from the supralabial gland and evolved independently three or four times in the family, always in lineages that invaded marine habitats. Our results suggest that the differentiated premaxillary glands are likely salt glands, as is the case in C. rynchops. If corroborated, this increases to six or seven the number of independent evolutionary origins of salt glands in snakes that have undergone an evolutionary transition to marine life.

In-depth immunorecognition and neutralization analyses of Micrurus mipartitus and M. dumerilii venoms and toxins by a commercial antivenom.

In Colombia, the Micrurus genus comprises 30 species, including M. mipartitus and M. dumerilii, which are of major clinical relevance due to their wide geographical distribution and the number of snakebites inflicted by them. These neurotoxic envenomations are characterized by neuromuscular paralysis attributed to venom components such as three-finger toxins (3FTx) and phospholipases (PLA2). Additionally, there is limited information available on the neutralizing coverage of commercially available antivenoms, underscoring the need to perform studies to assess the cross-neutralizing ability of these life-saving products. Therefore, we present an in-depth immunorecognition analysis by the anticoral-INS antivenom from Colombia on the M. mipartitus and M. dumerilii venoms. The antivenom cross-recognized the whole venoms and their components with different intensities. For instance, the antivenom showed better recognition on PLA2s than on 3FTxs in both venoms. Moreover, at doses tested, the antivenom totally neutralized the lethal effect of M. dumerilii venom; however, it did not neutralize this effect induced by M. mipartitus venom and its main toxic components from the southwestern region of the department of Antioquia. Furthermore, the anticoral-INS antivenom displayed better cross-immunorecognition of PLA2-predominant Micrurus venoms than of 3FTx-predominant Micrurus venoms. This highlights the need to include venoms from both types of venom patterns in the immunization mixture to produce antivenoms against coral snakes. Finally, our results suggest the need for further research to optimize the composition of immunizing mixtures for antivenom production and improve their efficacy against coral snake envenomation in Colombia and the Americas.

Ontogenetic and sexual differences in the venom of Bothrops moojeni: insights from a litter and its mother / Diferenças ontogenéticas e sexuais no veneno de Bothrops moojeni: percepções de uma ninhada e de sua mãe

Variability in snake venom composition is well-documented and crucial for understanding snake ecology and predicting snakebites. In this study, we characterize the venom composition and biological activities of newborn female and male Bothrops moojeni and their mother. Our results reveal significant differences between the venom of newborn females and males, demonstrating a broad and diverse range of proteins. The venoms of newborn females showed higher serine protease effects, increased hemorrhagic activity, and greater lethality compared to the venom of newborn males. However, no differences were observed in phospholipase A2 and coagulant activity. The differences in protein composition and toxic activities between maternal and neonatal venom, as well as between the venoms of newborn females and males, contribute to understanding the diverse outcomes of snakebites. These results underscore the importance of considering sex and ontogeny in understanding venom composition in snakes.

A variabilidade na composição dos venenos de serpentes é bem documentada e crucial para a compreensão da ecologia das serpentes ou do prognóstico dos envenenamentos ofídicos. Nesse estudo, caracterizamos a composição e a atividade biológica do veneno de fêmeas e machos recém-nascidos e sua mãe. Nossos resultados revelaram diferenças significativas entre o veneno de fêmeas e machos recém-nascidos, demonstrando uma ampla e diversificada gama de proteínas. Os venenos de fêmeas recém-nascidas apresentaram mais efeitos de serina protease, aumento da atividade hemorrágica e maior letalidade em comparação ao veneno dos machos recém-nascidos. No entanto, nenhuma diferença foi observada na fosfolipase A2 e na atividade coagulante. As diferenças na composição proteica e nas atividades tóxicas entre o veneno materno e neonatal, bem como entre os venenos de fêmeas e machos recém-nascidos contribuem para a compreensão dos diversos resultados dos acidentes ofídicos. Esses resultados ressaltam a importância de considerar o sexo e a ontogenia na compreensão da composição do veneno desses animais.

Habituação ou sensibilização? Efeito comportamental em Cascavel sul-americanas (Crotalus durissus)

The defensive behavior represents a crucial strategy for snakes, influenced by intrinsic factors that play significant roles in the survival, reproduction, and evolution of species. The present study focused on the rattlesnake (Crotalus durissus), a species exhibiting a wide variety of defensive behaviors. The research aimed to assess the manifestation of habituation or sensitization in the defensive behavior of rattlesnakes, establishing a comparison between adult and young individuals. For this purpose, 44 snakes (22 juveniles and 22 adults) kept in captivity were subjected to encounters with a white-eared opossum plush model in a controlled arena. Data collection extended over a period of 6 days, during which behaviors were recorded. The statistical analysis employed a mixed-effects generalized linear model (GLMM) with Poisson distribution, using R software (version 3.34). The results revealed an increase in aggression in both adult and juvenile groups on the sixth day. Notably, juveniles exhibited higher responsiveness and aggression compared to adults. This study provided clear evidence of ontogenetic differences in the selection of defensive strategies between juveniles and adults. This distinction was influenced by experience, limitations, and inherent needs at each life stage of the snakes. Contrary to expectations based on the literature, both juvenile and adult groups did not exhibit habituation to the presented stimuli, indicating instead a sensitization phenomenon. These findings emphasize the importance of considering ontogenetic variables when analyzing defensive behavior in snakes, particularly in the context of predator-prey interactions. Furthermore, the results have practical implications, especially in regions where the presence of rattlesnakes may pose risks to the community. This study contributes to a deeper understanding of defensive behavior patterns in rattlesnakes, providing valuable insights for management and conservation strategies in controlled environments.

O comportamento defensivo representa uma estratégia crucial para as serpentes, sendo influenciado por fatores intrínsecos que desempenham papéis significativos na sobrevivência, reprodução e evolução das espécies. O presente estudo concentrou-se na cascavel (Crotalus durissus), uma espécie que exibe uma ampla variedade de comportamentos defensivos. O objetivo da pesquisa consistiu em avaliar a manifestação de habituação ou sensibilização do comportamento defensivo em cascavéis, estabelecendo uma comparação entre indivíduos adultos e jovens. Para tanto, foram empregadas 44 serpentes (22 filhotes e 22 adultos) mantidas em cativeiro, submetidas a confrontos com um modelo de pelúcia de gambá-de-orelha-branca em uma arena controlada. A coleta de dados estendeu-se por um período de 6 dias, durante os quais foram registrados os comportamentos. A análise estatística adotada foi conduzida por meio de um modelo linear generalizado misto (GLMM) com distribuição de Poisson, utilizando o Software R (versão 3.34). Os resultados revelaram um aumento na agressividade tanto nos grupos de adultos quanto de filhotes no sexto dia. Destacou-se que os filhotes apresentaram maior responsividade e agressividade em comparação com os adultos. Este estudo proporcionou evidências claras de uma diferença ontogenética na escolha de estratégias defensivas entre filhotes e adultos. Tal distinção foi influenciada pela experiência, limitações e necessidades inerentes a cada fase de vida das serpentes. Contrariando as expectativas baseadas na literatura, ambos os grupos, filhotes e adultos, não manifestaram habituação aos estímulos apresentados, indicando, em vez disso, um fenômeno de sensibilização. Esses achados enfatizam a relevância de considerar as variáveis ontogenéticas ao analisar o comportamento defensivo em serpentes, particularmente no contexto das interações presa-predador. Além disso, os resultados possuem implicações práticas, especialmente em regiões onde a presença de cascavéis pode representar riscos para a comunidade. Este estudo contribui para uma compreensão mais aprofundada dos padrões de comportamento defensivo em cascavéis, fornecendo insights valiosos para estratégias de manejo e conservação em ambientes controlados.

Atividade diária, uso do ambiente e comportamento reprodutivo de Bothrops sazimai, espécie criticamente ameaçada, em cativeiro

The genus Bothrops, popularly know as the jararacas group, currently has 32 species in Brazil. They are found both at the sea level and at altitudes close to 3000m, and some species unhabitat continental islands. One of these species is Bothrops sazimai, described in 2016, a snake endemic to Ilha dos Franceses which, due to anthropogenic pressure in its natural environment, is considered a critically endangered species. Therefore, maintaning this speciesw in captivity can help to understand its behavior, biology and environmental use. In this work, through monitoring over 15 days, it was possible to characterize the daily activity, environmental use and reproductive behavior of two individuals of B. sazimai, and conclude that the species presentes nocturnal activity and semi-arboreal habits, despite using the lower stratum more frerquently. This work also presentes aspects of reproductive behavior in captivity, showing that the courtship behavior of B. sazimai is similar to the behavior of other snakes of the genus Bothrops, which basically involves a phase of pursuit, alignment and introduction of the hemipenis. To date, the information available on B. sazimai is limited to the work describing the species, therefore, the information obtained through this work is unprecedented.

O gênero Bothrops, conhecido popularmente como grupo das jararacas apresenta atualmente, no Brasil, 32 espécies. São encontradas tanto ao nível do mar, quanto em altitudes próximas a 3000m, e algumas espécies habitam ilhas continentais. Uma dessas espécies é a Bothrops sazimai, descrita em 2016, serpente endêmica da Ilha dos Franceses que, devido à pressão antropogênica em seu ambiente natural, é considerada uma espécie criticamente ameaçada. Dessa forma, a manutenção dessa espécie em cativeiro pode auxiliar no conhecimento do comportamento, biologia e uso do ambiente. Neste trabalho, através do monitoramento ao longo de 15 dias, foi possível caracterizar a atividade diária, uso do ambiente e comportamento reprodutivo de dois indivíduos de B. sazimai, e concluir que a espécie apresenta atividade noturna e hábitos semi arborícolas, apesar de usar o estrato inferior com mais frequência. Este trabalho também apresenta aspectos do comportamento reprodutivo em cativeiro, mostrando que o comportamento de corte de B. sazimai é semelhante ao comportamento de outras serpentes do gênero Bothrops, que envolve basicamente a fase de perseguição, alinhamento e introdução do hemipenis. Até o momento, as informações disponíveis sobre B. sazimai limitam-se ao trabalho de descrição da espécie, sendo assim, as informações obtidas por este trabalho são inéditas.

Stable isotopic composition of Bothrops atrox (common lancehead) in four distinct land covers in eastern Amazonia

Since consumers reflect the isotopic composition of an assimilated diet, stable isotopes can be a useful tool to address the feeding ecology of tropical snakes. This is the first study reporting carbon and nitrogen stable isotopic composition of Bothrops atrox (Linnaeus, 1758) living in different landscapes located in the lower Amazon river, encompassing four main natural landscapes of the Amazon: old-growth forests, várzeas (flooded forests), savannas, and pastures. Our null hypothesis is that the δ13C of forest specimens of B.atrox is more negative because forests are dominated by C3 plants, while C4 plants are common in the other landscapes. On the other hand, δ15N of forest specimens should be more positive, since the δ15N of old-growth forests are higher than plants of savanna, várzea, and pastures. Confirming our hypothesis, the δ13C of B. atrox scales of the Tapajós National Forest was approximate −25‰ to −24‰, increased to approximately −23.5‰ to −23.0‰ in the savanna and pasture, and to −21‰ in the várzea, showing an increased contribution of C4-derived carbon. Some specimens of B. atrox had δ15N as high as 18‰, which is much higher than the average δ15N of the snake's prey (7‰), confirming the apex position of B. atrox in the Amazon region. The δ15N values of the forest specimens were 5‰ higher than the savanna specimens, and this difference decreased to 3‰ between the forest and the pasture, and the várzea specimens. Finally, there were not large differences between δ15N values of livers and scales in any of the landscapes, suggesting a constant diet through time, and reinforcing the possibility of the use of snake's scale as a less invasive and non-lethal tissue to analyze.

Diagnosis of endoparasite species and subtypes of Cryptosporidium spp. with one health importance, in feces from captive snakes in Rio de Janeiro, Brazil.

In captivity, snakes may present chronic infections with high mortality, such as those caused by Cryptosporidium serpentis, or they may be pseudoparasitized by species that present zoonotic potential. Thus, the aim of this study was to evaluate the frequency of helminths and protozoa in the feces of captive snakes, characterize the species and subtypes of Cryptosporidium spp. and correlate the parasites detected with other information obtained from these animals. Feces were collected from 189 snakes kept at the Vital Brazil Institute, Rio de Janeiro, including samples from Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops atrox, Bothrops leucurus, Crotalus durissus and Lachesis muta. All the samples were subjected to microscopy techniques and the polymerase chain reaction (PCR) in association with sequencing, to identify Cryptosporidium spp.. Forms of parasites infecting the snakes were identified through microscopy in 50.8% of the samples. Helminths were detected more often than protozoa in the feces of these animals, mainly comprising eggs resembling Kalicephalus sp. and oocysts of Eimeria sp.. Pseudoparasites such as Syphacia sp., Aspiculuris sp. and Hymenolepis nana were also detected. Through correlating the results obtained from parasitological staining techniques and PCR, the total frequency of Cryptosporidium sp. was found to be 19%. The species C. tyzzeri and C. parvum were identified. Characterization using the target gp60 showed subtypes with high potential for zoonotic transmission, especially IIaA15G2R1 and IIaA14G2R1 of C. parvum and IXbA8 of C. tyzzeri. This study highlighted the need for more intensive health management in the Institute's serpentarium and, especially, in its bioterium where rodents are reared as a food source for these snakes.

Patterns of morphological variation and ecological correlates in the skull of vipers (Serpentes: Viperidae).

The skull of vipers is a highly kinetic anatomical structure involved in envenomating and consuming of prey. Morphological knowledge about the viperid skull is based on studies on some groups of species, but information on its variation within the whole family and its functional morphology is still scarce. In this study, we aimed to explore variation in skull morphology among species of the three subfamilies of Viperidae, and test whether that variation correlates with macrohabitat and diet. We performed quantitative analyses of the viperid skull based on broad taxonomic sampling and two methodological approaches: linear and geometric morphometrics. The results of both approaches showed that much of the variation lies in differences of shape and relative size of the premaxilla, the nasals, the frontals, and the parietals. The results indicated that phylogeny and size influence the shape of the skull, but we also found evidence of morphological differentiation between arboreal and terrestrial species and in species with mammal specialist diet. Our findings imply that, besides evolutionary allometry and phylogenetic signal, demands of particular diets coupled with use of certain habitats have in part shaped morphological evolution of the viperid skull.

STENOPHYSA MARMORATA (MOLLUSCA: PHYSIDADE) AS AN INTERMEDIATE HOST OF A SNAKE TREMATODE OF THE FAMILY RENIFERIDAE IN BRAZIL.

Species of the family Reniferidae are trematodes found in the oral cavity and esophagus of snakes from Nearctic and Neotropical regions. Although Renifer heterocoelium has been reported in different snake species from South America, the snails involved in its transmission remain unknown. In this study, a xiphidiocercaria emerged from the physid snail Stenophysa marmorata from Brazil and was subjected to morphological and molecular study. The general morphology, including the shape of the stylet and arrangement of penetration glands, resembles that described for reniferid trematodes from North America. Phylogenetic analysis on the basis of nuclear sequences (28S ribosomal deoxyribonucleic acid gene [1,072 base pairs {bp}] and internal transcribed spacer region [ITS, 1,036 bp]) supports identifying this larva as a member of the family Reniferidae, very possibly a species of the genus Renifer. In the 28S analysis, low molecular divergences were found to Renifer aniarum (1.4%) and Renifer kansensis (0.6%), but also concerning other 2 reniferid species, i.e., Dasymetra nicolli (1.4%) and Lechriorchis tygarti (1.0%). Regarding ITS, the divergences between this Brazilian cercaria and R. aniarum or L. tygarti were 1.9% and 8.5%, respectively. In the case of the mitochondrial marker cytochrome oxidase subunit 1 (797 bp), our Reniferidae gen. sp. differs 8.6-9.6% from Paralechriorchis syntomentera, the only reniferid with sequences available for comparison. We discuss the probable conspecificity of the larval stages here reported with R. heterocoelium, the reniferid species reported in South America.

Effect of proteins isolated from Brazilian snakes on enterovirus A71 replication cycle: An approach against hand, foot and mouth disease.

Enterovirus A71 (EVA71) belongs to the Picornaviridae family and is the main etiological agent of hand, foot, and mouth disease (HFMD). There is no approved antiviral against EVA71, and therefore the search for novel anti-EVA71 therapeutics is essential. In this context, the antiviral activity of proteins isolated from snake venoms has been reported against a range of viruses. Here, the proteins CM10 and CM14 isolated from Bothrops moojeni, and Crotamin and PLA2CB isolated from Crotalus durissus terrificus were investigated for their antiviral activity against EVA71 infection. CM14 and Crotamin possessed a selective index (SI) of 170.8 and 120.4, respectively, while CM10 and PLA2CB had an SI of 67.4 and 12.5, respectively. CM14 inhibited all steps of viral replication (protective effect: 76 %; virucidal: 99 %; and post-entry: 99 %). Similarly, Crotamin inhibited up to 99 % of three steps. In contrast, CM10 and PLA2CB impaired one or two steps of EVA71 replication, respectively. Further dose-response assays using increasing titres of EVA71 were performed and CM14 and Crotamin retained functionality with high concentrations of EVA71 (up to 1000 TCID50). These data demonstrate that proteins isolated from snake venom are potent inhibitors of EVA71 and could be used as scaffolds for future development of novel antivirals.

Morbidity survey of the history of snakebites in different communities in the alto Juruá, western Brazilian Amazon.

The Alto Juruá region has a rich diversity of venomous snakes, and snakebites represent an important cause of morbidity. The present study was carried out in five communities in the Alto Juruá region, and aimed to evaluate the history of snakebites in terms of which snakes were involved and what was the victims' conduct in relation to first aid and prevention. Between 2017 and 2019, semi-structured interviews were applied to 260 residents, 56 (21.53%) of whom had a history of snakebites. The profile of the victims is 73.2% male, a resident of rural areas who develops activities mainly in agriculture and extractivism and who has a low level of education. Just over half of the snakebites occurred during the rainy season (52.8%) and the majority during the day (77%). Bothrops bites were the most frequently reported (90.3%) and B. atrox was the most mentioned snake (jararaca and surucucu) for being responsible for snakebites (80.8%). Approximately 87.5% of the bites occurred to the lower limbs, and 48.2% of the victims were barefoot at the time of the bite. Inadequate first-aid procedures were reported, such as using tourniquets and consuming the drink known as "Específico Pessoa", not drinking water, and treatment with traditional therapy (phytotherapics, zootherapeutics and faith healers), as well as not seeking hospital care. Thus, the need to facilitate access to hospital units in these communities and health education (prevention and first aid) are essential in order to minimize this situation.

Envenomation by Micrurus hemprichii in Brazilian Amazonia: A report of three cases.

Herein, we report three new separate cases of human envenomations by Micrurus hemprichii for the Amazon, which is a biome where envenomations by Micrurus are seldom reported. Two women were bitten after stepping on the snakes and one man was bitten while handling the animal. All cases occurred in the peridomicile, in rural areas. The first case evolved mainly to local symptoms, but the patient was discharged before the identification of the snake and had to be called back for observation before being eventually discharged. In the second case, the patient presented transitory dyspnea and she was discharged after four days in hospital. In the third case, the patient showed only local symptoms, but he was about to receive unnecessary antivenom against coral snakes. Cases like these show the importance of educational problems regarding local venomous snakes in order to avoid bites and to provide the correct hospital treatment. For this, trained professionals in cases involving venomous animals are needed.

Antibodies against a single fraction of Micrurus dumerilii venom neutralize the lethal effect of whole venom.

The coralsnake Micrurus dumerilii (Elapidae) is reported to cause envenomings of medical importance. Previous studies characterized the protein composition of its venom, with phospholipase A2 (PLA2) proteins the most abundant. However, it is unknown which venom components are responsible for its lethal toxicity. Fractionation of M. dumerilii venom from Colombia was carried out using RP-HPLC and each fraction was screened for lethal effect in mice at a dose of 20 µg by intraperitoneal route. Results showed that only one fraction, F9, was lethal. This fraction displayed PLA2 activity, induced indirect hemolysis in vitro, as well as edema and myotoxicity in vivo. SDS-PAGE of unreduced F9 evidenced two bands of 8 and 15 kDa, respectively, consistent with the detection of proteins with masses of 13,217.77 Da, 7144.06 Da, and 7665.55 Da. Tryptic digestion of F9 followed by nESI-MS/MS revealed peptide sequences matching proteins of the three-finger toxin (3FTx) and PLA2 families. Immunization of a rabbit with F9 proteins elicited antibody titers up to 1:10,000 by ELISA. After serum fractionation with caprylic acid, the obtained IgG was able to neutralize the lethal effect of the complete venom of M. dumerilii using a challenge of 2 ×LD50 at the IgG/venom ratio of 50:1 (w/w). In conclusion, present results show that the lethal effect of M. dumerilii venom in mice is mainly driven by one fraction which contains 3FTx and PLA2 proteins. The antibodies produced against this fraction cross-recognized other PLA2s and neutralized the lethal effect of whole M. dumerilii venom, pointing out to the potential usefulness of F9 as a relevant antigen for improving current coral snake antivenoms.

Two records of xanthism in Corallus hortulana (Serpentes: Boidae) in Bolivia with comments on the yellow, patternless morphotype / Dos registros de xantismo en Corallus hortulana (Serpentes: Boidae) en Bolivia con comentarios sobre el morfotipo amarillo sin patrón

Snakes display a wide range of skin colors and patterns, acting in ecological, behavioral, and physiological roles. Xanthism has been reported in numerous snake species, including many neotropical taxa. Corallus hortulana is a polychromatic species widely distributed throughout South America. Yellow, patternless individuals, while rare, have been recognized as within the species' polychromatic range. We report the first yellow, patternless specimens of Corallus hortulana collected in Bolivia and discuss whether these individuals fit the criteria of xanthism.(AU)

Las serpientes muestran una amplia gama de colores y patrones de piel, actuando en roles ecológicos, de comportamiento y fisiológicos. Además, se ha informado xantismo en numerosas especies de serpientes, incluidos muchos taxones neotropicales. Corallus hortulana es una especie policromática ampliamente distribuida en América del Sur. Los individuos amarillos sin patrón, aunque raros, se han reconocido dentro del rango policromático de la especie. Reportamos los primeros especímenes amarillos sin patrón de Corallus hortulana recolectados en Bolivia y discutimos si estos individuos cumplen con los criterios de xantismo.(AU)

Comparing morphological and secretory aspects of cephalic glands among the New World coral snakes brings novel insights on their biological roles

Oral and other cephalic glands have been surveyed by several studies with distinct purposes. Despite the wide diversity and medical relevance of the New World coral snakes, studies focusing on understanding the biological roles of the glands within this group are still scarce. Specifically, the venom glands of some coral snakes were previously investigated but all other cephalic glands remain uncharacterized. In this sense, performing morphological and molecular analysis of these glands may help better understand their biological role. Here, we studied the morphology of the venom, infralabial, rictal, and harderian glands of thirteen species of Micrurus and Micruroides euryxanthus. We also performed a molecular characterization of these glands from selected species of Micrurus using transcriptomic and proteomic approaches. We described substantial morphological variation in the cephalic glands of New World coral snakes and structural evidence for protein-secreting cells in the inferior rictal glands. Our molecular analysis revealed that the venom glands, as expected, are majorly devoted to toxin production, however, the infralabial and inferior rictal glands also expressed some toxin genes at low to medium levels, despite the marked morphological differences. On the other hand, the harderian glands were dominated by the expression of lipocalins, but do not produce toxins. Our integrative analysis, including the prediction of biological processes and pathways, helped decipher some important traits of cephalic glands and better understand their biology.

Utilização de ensaios moleculares na detecção de vírus em serpentes dos plantéis do Instituto Butantan

Viruses can infect and cause diseases in snakes, causing outbreaks in herds and hindering the production of venom. Reptarenavirus were identified in cases of Inclusion Body Disease, and substantial evidence was presented suggesting the causal relationship of the virus with the disease, including co-infection of different virus species in the same animal. Chuviridae is a recently discovered family of negative-sense, single-stranded RNA viruses of variable genomic structure that infect several species of arthropods, including mosquitoes. This work was carried out as part of the implementation of methodologies for detection and identification of viruses in snakes kept in captivity in the herds of Instituto Butantan, which aim to optimize and improve protocols for handling and conservation of animals, in addition to the possibility of identifying new virus species. Sample collections were performed in vivo and/or post-mortem, considering the clinical situation of each animal and at the discretion of the veterinarian responsible for the vivarium. After extracting the nucleic acids, virus research was performed using the amplification of viral genome fragments by polymerase chain reaction and nucleotide sequencing by the Sanger method. The methodologies developed in this work were effective in detecting reptarenavirus and chuvirus in snakes kept in captivity at Instituto Butantan, which demonstrates the importance of implementing the diagnosis of viral diseases in the quarantine and sanitary surveillance of these collections.

Vírus podem infectar e causar doenças em serpentes, ocasionando surtos em biotérios e consequente comprometimento na produção de veneno. Reptarenavírus foram identificados em casos de Doença do Corpúsculo de Inclusão, sendo apresentadas evidências substanciais sugerindo a relação causal do vírus com a doença, inclusive com a coinfecção de diferentes espécies de vírus em um mesmo animal. Chuviridae é uma família recentemente descoberta de vírus RNA de fita simples, de sentido negativo e de estrutura genômica variável, que infecta várias espécies de artrópodes, incluindo mosquitos. Este trabalho foi realizado como parte da implantação de metodologias de detecção e identificação de vírus em serpentes mantidas em cativeiro nos plantéis do Instituto Butantan, que visam a otimização e o aperfeiçoamento dos protocolos de manejo e conservação dos animais, além da possibilidade de identificação de novas espécies de vírus. As coletas de amostras foram realizadas in vivo e/ou post-mortem, considerando a situação clínica de cada animal e a critério do médico veterinário responsável pelo biotério. Após a extração dos ácidos nucleicos, a pesquisa de vírus foi realizada utilizando a amplificação de fragmentos do genoma viral por reação em cadeia da polimerase e sequenciamento de nucleotídeos pelo método de Sanger. As metodologias desenvolvidas neste trabalho foram eficazes na detecção de reptarenavírus e chuvírus em serpentes mantidas em cativeiro em plantéis do Instituto Butantan, o que demonstra a importância da implementação do diagnóstico de doenças virais na quarentena e no monitoramento sanitário dessas coleções.

Determinação do index corporal de serpentes do gênero Bothrops

The Laboratory of Herpetology at Butantan Institute maintains approximately one thousand specimens of venomous snakes in its serpentarium, with the main purpose of producing immunobiologicals and for various basic and applied research. In order to obtain venom with good quality, it is essential the healthiness of all snakes.To monitor the health of snakes, hematological, biochemical and ultrasound exams areperformed. All snakes belonging to the serpentarium have an Individual Form, in whichthe monthly weight and the rostrum-cloacal length (RCL) and total length (TL) are noted, among other relevant observations. However, there is still difficulty in determining the ideal weight of different species of snakes in captivity. Some works report the use of palpation to assess the body condition of the snakes, giving a score ranging from 1 to 5, however this is a subjective method and requires physical restraintof the animal. Other works calculate the body index of snakes through formulas usingthe animal's mass; these formulas are usually extrapolated from mammals. In our work,we used the relationship between the mass and the CRC of several individuals recentlyarrived from nature and healthy, from the five species of Bothrops used for the production of Bothropic antiserum. We used male and female snakes of the species Bothrops jararaca, B. jararacussu, B. neuwiedi Complex, B. moojeni and B. alternatus.Through the linear regression obtained by the natural log of CRC and the natural log of mass, an equation with significant p and r2 values for all studied species was generated. With the aid of this equation, it was possible to determine the estimated weight, the minimum weight and the maximum weight of the five different species and,in this way, monitor the body condition of snakes kept in captivity through the mass and CRC recorded in the Individual Form. Only snakes under the minimum weight or above the maximum weight will be contained for clinical examination and appropriate treatment.

O Laboratório de Herpetologia do Instituto Butantan mantém em seu plantel aproximadamente mil espécimes de serpentes peçonhentas, tendo como principal finalidade a produção de imunobiológicos e a utilização para diversas pesquisas básicas e aplicadas. Para a obtenção de veneno de boa qualidade é fundamental queas serpentes estejam hígidas. Para monitorar a saúde das serpentes são realizados exames hematológicos, bioquímicos e ultrassonográficos. Todas as serpentes do biotério possuem uma Ficha Individual, em que são anotados o peso mensal e o comprimento rostro-cloacal (CRC) e comprimento total (CT) semestral, entre outras observações relevantes. Contudo, ainda existe dificuldade em se determinar qual o peso ideal das diferentes espécies de serpentes em cativeiro. Alguns trabalhos relatam o uso da palpação para avaliar a condição corporal da serpente, dando um escore que varia de 1 a 5, contudo este é um método subjetivo e que necessita a contenção física do animal. Outros trabalhos calculam o index corporal das serpentesatravés de fórmulas utilizando a massa do animal; fórmulas estas extrapoladas de mamíferos. No nosso trabalho utilizamos a relação entre a massa e o CRC de vários indivíduos recém-chegados da natureza e aparentemente saudáveis das cinco espécies de Bothrops utilizadas para a produção do soro antibotrópico. Foram utilizadas serpentes machos e fêmeas das espécies Bothrops jararaca, B.jararacussu,Complexo B. neuwiedi, B. moojeni e B. alternatus. Através da regressão linear obtida através do Log natural do CRC e Log natural da massa foi gerada uma equação da reta com valores de p e r2 significativos para todas as espécies estudadas.Com o auxílioda equação gerada neste estudo foi possível determinar o peso estimado, o peso mínimo e o peso máximo das diferentes espécies e, deste modo, monitorar a condiçãocorporal das serpentes mantidas em cativeiro através da massa e CRC anotados nas Fichas Individuais. Apenas as serpentes com pesos abaixo do peso mínimo ou acimado peso máximo serão contidas para o exame clínico e tratamento adequado.