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Este inventário foi desenvolvido na Serra de Piquiatuba, inserida na área do 8º Batalhão de Engenharia e Construção (8º BEC), localizado no km 8 da rodovia Santarém–Cuiabá, no município de Santarém, Pará, Brasil. Esta área é caracterizada por um clima quente e úmido, pluviosidade acima de 1900 mm/ano e um alto grau de preservação. As coletas foram realizadas no período diurno (manhã), mensalmente, durante um ano, de julho/2011 a junho/2012, utilizando o guarda-chuva entomológico para amostragem. Foram obtidas 71 amostras, que resultaram na captura de 917 aranhas, distribuídas em 23 famílias e 74 espécies (409 adultos, [44,6%] e 508 imaturos [55,4%]). A maior abundância de aranhas foi na estação chuvosa (dezembro a junho). As famílias Tetragnathidae, Araneidae, Salticidae, Theridiidae e Uloboridae foram as mais abundantes. Araneidae apresentou maior riqueza, com 25 espécies seguida de Theridiidae (11 espécies) e Salticidae (8 espécies). Após um ano de coletas, a assíntota da curva de acumulação não foi atingida, sendo a menor estimativa de 85.18 (Chao 1) e a maior com 110.88 (Jack 2), mostrando que mais investimento em amostragem devem ser realizados. Esta é a primeira listagem de aranhas de uma área da região oeste do Pará, abrangendo uma área preservada e protegida, como a Serra de Piquiatuba, na área do 8° BEC.

Raimundo Nonato Colares Camargo Júnior,1,&ast; Jaciara Simone Sarmento Gomes,2,&ast; Mônica Cristina Corrêa Carvalho,2,&ast; Hipócrates de Menezes Chalkidis,2,&ast; Welligton Conceição da Silva,1,&ast; Juliana Sousa da Silva,3,&ast; Samia Rubielle Silva de Castro,4,&ast; Raul Cunha Lima Neto,4,&ast; Victor Hugo Pereira Moutinho4,&ast; 1Animal Science Graduate Program, Federal University of Pará (UFPA), Belém, Pará, Brazil; 2Department of Biological and Health Sciences, Amazon University Center (Unama), Santarém, Pará, Brazil; 3Postgraduate Program in Natural Resources of the Amazon, Federal University of Western Pará, Santarém, Pará, Brazil; 4Institute of Biodiversity and Forests, Federal University of Western Pará, Santarém, Pará, Brazil&ast;These authors contributed equally to this workCorrespondence: Raimundo Nonato Colares Camargo Júnior, Federal University of Pará (UFPA), R. Augusto Corrêa, 01 - Guamá, Belém, Pará, 66075-110, Brazil, Tel +55 015 93 98124-9425, Fax +55 015 93 98124-9425, Email camargojunior@gmail.comIntroduction: Human visceral leishmaniasis (VL) is a zoonosis of great importance to public health due to its epidemiological diversity, with emphasis on the possibility of aggravation by coinfection with the human immunodeficiency virus (HIV).Objective: The aim was to study the epidemiological characteristics of VL cases associated with HIV coinfection in Pará. Methods. Reported cases of VL from January 2006 to December 2016 were investigated. A descriptive epidemiological method related to age, gender, area of residence and coinfection with HIV was used. To calculate variance and test equity, the F-test (Fisher) was performed. To observe the influence of one aspect on another, the chi-square was used to verify if there was dependence or independence between the variables.Results: A total of 1171 cases of VL were reported during the study period. There was an annual mean of LV of 94.9, with a statistical difference (p< 0.05) between age groups, with the highest number of cases being observed in children aged 1 to 4 years (27.16%). Males and the urban area had a higher number of cases. There were 57 cases of VL/HIV coinfection, with emphasis on the year 2013 and the municipality of Santarém, which had the highest number of cases. During the ten years studied, there was a correlation between coinfection VL/ HIV, with significant differences between patients with and without HIV who contracted VL (p< 0.001).Conclusion: The data reveal the endemic nature of VL in the region, with a high percentage of infection in children living in urban areas. Although the studied region is not identified as a predominant area of HIV cases, this study showed a high annual average (10.3) of cases of VL/HIV coinfection being the first time that cases of VL/HIV coinfection were reported in the Mesoregion of the Lower Amazon and Southwest Pará.Keywords: age group, male, children, analytical epidemiology

Snakebite envenomation is considered a neglected public health problem in Brazil. Most of such accidents are attributed to different species of the genus Bothrops, of which Bothrops atrox is responsible for most cases. Snake venoms consist primarily of proteins, each of which performs a biological activity or acts synergetically to help snakes to survive by capturing and digesting prey and to defend themselves against attacks by predators. Studies have reported intraspecific and intrapopulational variability that depends on the age (ontogenetic variation), sex and size of the snake as well as geographic region. In the present study, variations in venom produced by the left and right glands of the same individual were assessed. Phospholipase A2 (PLA2) and coagulant activities and protein profiles were analyzed, and the immunogenic proteins in venom from each gland were titrated. In vitro results for venom from seven snakes of the species B. atrox (14 samples; one from each gland of seven snakes) revealed a statistically significant difference in PLA2 activity and coagulant activity between venom from the left and right glands in one snake. In addition, minor differences in protein profile were observed in SDS-PAGE and in the minimum concentration of proteins reactive to a polyclonal antiserum against B. atrox venom in a Dot-ELISA. The results therefore suggest that the same snake may be able to produce biochemically distinct venoms. However, the frequency with which these variations occur, their extent and whether they can cause clinical differences in vivo are not known.

A Vigilância Epidemiológica (VE) tem como papel principal manter segura as condições de vida da sociedade. Por meio dela, é possível identificar e tratar casos clínicos que afetam a saúde da população. Neste trabalho, foi possível exemplificar o papel íntegro e eficaz desse órgão. A metodologia utilizada inclui a busca e análise de dados endêmicos que, de forma brusca ou sutil, geram consequências na comunidade. Desta forma, o trabalho apresentado propôs uma revisão de literatura sobre a relevância das ações mediadas pela VE, abordando suas ações profiláticas e também se referindo a forma de atuação desde o momento em que a comunidade apresenta casos suspeitos de doenças, até que os casos sejam sanados.

Objetivou-se estimar o prejuízo econômico oriundo de carcaças de bovinos e bubalinos condenados por tuberculose, durante o ano de 2018, na microrregião de Santarém, na mesorregião do Baixo Amazonas, no estado do Pará, Amazônia Oriental. Os dados foram fornecidos pela Secretaria Municipal de Agricultura e Pesca, no setor de inspeção municipal, Santarém, Pará. Estes continham informações de três abatedouros frigoríficos sobre Serviço de Inspeção Municipal, sobre a espécie bovina e bubalina, especificamente: sexo e o total de animais abatidos e condenados. Foram inspecionados 23.963 bovinos e bubalinos no ano de 2018, provenientes de municípios da mesorregião do Oeste do Pará. Calculou-se também a prevalência de condenação e os prejuízos econômicos. Ao todo foram condenadas por tuberculose 74 carcaças de bovinos e bubalinos. Desse quantitativo, 43 eram fêmeas e 31 machos. A prevalência foi de 0,006%. As perdas econômicas apresentadas em ordem decrescente devido à condenação de carcaça no rebanho, para fêmeas e para machos, foram equivalentes a R$ 293.040,00; R$ 170.280,50 e R$ 122.760,00, respectivamente. Conclui-se que a prevalência foi baixíssima para a tuberculose e o prejuízo econômico decorrente desta enfermidade foi significativo, necessitando-se investigar o prejuízo econômico que as diversas enfermidades causam no Baixo Amazonas para mitigar as perdas futuras decorrentes desta e de outras enfermidades.

Variability in the composition of snake venoms occurs in different taxa and is usually correlated to snake fitness. Here, we compared B. atrox venoms from three different geographic regions across the Brazilian Amazon and found remarkable functional differences particularly between venoms from two populations separated by the Amazon River, in specimens born, raised and maintained under the same conditions at Instituto Butantan serpentary. Venom from Presidente Figueiredo snakes induced stronger dermonecrosis, but was less procoagulant and lethal to mice; these activities were correlated to the presence of a PI-class SVMP and absence of a SVSP in the venom, respectively. Venom from São Bento snakes was more hemorrhagic, killed mice more efficiently, but induced lower signs of dermonecrosis, which was correlated to the higher proportion of SVMPs and the absence of a PI-class SVMP isoform. Belterra snakes, a reference of wild snakes, presented venoms with intermediate phenotypes. Commercial Bothrops antivenom was effective in neutralizing all biological activities evaluated in this study, including dermonecrosis and pro-coagulant, which are relevant for human snakebite accidents by B. atrox. Functional differences correlated to snake fitness may also imply in different symptomatology for B. atrox snakebite patients and deserve special attention from clinical toxicologists.

The use of natural substances from animals and plants has always been the subject of research, because it is made in a rustic way and through popular knowledge, built by everyday experiences and without scientific approval. The oil from the body fat of the snake - or better known as lard of anaconda, extracted from the anaconda snake (Eunectes murinus) is a product widely used in traditional communities and is marketed in markets for therapeutic purposes in several diseases. Because of the low cost and popular beliefs, many people use zootherapeutic means to treat themselves. Hence the importance of therapeutic proof of these products. Thus, the aim of this study was to verify whether the lard of anaconda made in the traditional way has antimicrobial potential in certain strains of pathogenic bacteria and fungi. For that, the evaluation of the lard’s antimicrobial potential was carried out using the disk diffusion method, adapted from the manual M7-A6 standardized by the Clinical and Laboratory Standards Institute, against the strains of S. aureus, E. coli, Ç. albicans and C. krusei. The observed results showed that there was no presence of bacterial or yeast growth inhibition halos, indicating that the anaconda lard oil did not inhibit the growth of fungi and bacteria in any of the strains used. Therefore, the present study allows us to conclude that the anaconda lard oil (Eunectes murinus), did not present any inhibitory antimicrobial potential, against the tested bacterial and fungal strains. O uso de substâncias naturais provenientes de animais e plantas sempre foi alvo de pesquisas, por ser feito de forma rustica e através de saberes populares, construídos por experiências no cotidiano e sem aprovação científica. O óleo da gordura corporal da serpente– ou mais conhecida como banha de sucuriju, extraída da serpente sucuri (Eunectes murinus) é um produto muito utilizado em comunidades tradicionais e é comercializada em mercados com finalidade terapêutica em diversas moléstias. Por conta do baixo custo e de crenças populares, muitas pessoas utilizam de meios zooterápicos para se tratar por si mesmo. Daí a importância da comprovação terapêutica desses produtos. Assim, o objetivo neste estudo foi verificar se a banha de sucuriju confeccionada de modo tradicional possui potencial antimicrobiano, em determinadas cepas de bactérias e fungos patogênicos. Para isso a avaliação do potencial antimicrobiano da banha, foi realizada através do método de difusão em disco, adaptado do manual M7-A6 padronizado pelo Clinicaland Laboratory Standards Institute, frente às cepas de S. aureus, E. coli, C. albicans e C. krusei. Os resultados observados demonstraram que não houve a presença de halos de inibição de crescimento bacteriano ou leveduriformes, indicando que o óleo da banha de sucuriju não inibiu o crescimento de fungos e bactérias em nenhuma das cepas utilizadas. Portanto, o presente estudo permite concluir que o óleo da banha de sucuriju (Eunectes murinus), não apresentou nenhum potencial antimicrobiano inibitório, frente às cepas bacterianas e fúngicas testadas. &nbsp

Differences in snake venom composition occur across all taxonomic levels and it has been argued that this variation represents an adaptation that has evolved to facilitate the capture and digestion of prey and evasion of predators. Bothrops atrox is a terrestrial pitviper that is distributed across the Amazon region, where it occupies different habitats. Using statistical analyses and functional assays that incorporate individual variation, we analyzed the individual venom variability in B. atrox snakes from four different habitats (forest, pasture, degraded area, and floodplain) in and around the Amazon River in Brazil. We observed venom differentiation between spatially distinct B. atrox individuals from the different habitats, with venom variation due to both common (high abundance) and rare (low abundance) proteins. Moreover, differences in the composition of the venoms resulted in individual variability in functionality and heterogeneity in the lethality to mammals and birds, particularly among the floodplain snakes. Taken together, the data obtained from individual venoms of B. atrox snakes, captured in different habitats from the Brazilian Amazon, support the hypothesis that the differential distribution of protein isoforms results in functional distinctiveness and the ability of snakes with different venoms to have variable toxic effects on different prey.

Venom variability is commonly reported for venomous snakes including Bothrops atrox. Here, we compared the composition of venoms from B. atrox snakes collected at Amazonian conserved habitats (terra-firme upland forest and varzea) and human modified areas (pasture and degraded areas). Venom samples were submitted to shotgun proteomic analysis as a whole or compared after fractionation by reversed-phase chromatography. Whole venom proteomes revealed a similar composition among the venoms with predominance of SVMPs, CTLs, and SVSPs and intermediate amounts of PLA2s and LAAOs. However, when distribution of particular isoforms was analyzed by either method, the venom from varzea snakes showed a decrease in hemorrhagic SVMPs and an increase in SVSPs, and procoagulant SVMPs and PLA2s. These differences were validated by experimental approaches including both enzymatic and in vivo assays, and indicated restrictions in respect to antivenom efficacy to variable components. Thus, proteomic analysis at the isoform level combined to in silico prediction of functional properties may indicate venom biological activity. These results also suggest that the prevalence of functionally distinct isoforms contributes to the variability of the venoms and could reflect the adaptation of B. atrox to distinct prey communities in different Amazon habitats. Biological significance In this report, we compared isoforms present in venoms from snakes collected at different Amazonian habitats. By means of a species venom gland transcriptome and the in silico functional prediction of each isoform, we were able to predict the principal venom activities in vitro and in animal models. We also showed remarkable differences in the venom pools from snakes collected at the floodplain (varzea habitat) compared to other habitats. Not only was this venom less hemorrhagic and more procoagulant, when compared to the venom pools from the other three habitats studied, but also this enhanced procoagulant activity was not efficiently neutralized by Bothrops antivenom. Thus, using a functional proteomic approach, we highlighted intraspecific differences in B. atrox venom that could impact both in the ecology of snakes but also in the treatment of snake bite patients in the region.

Variability in snake venom composition has been frequently reported and correlated to the adaptability of snakes to environmental conditions. Previous studies report plasticity for the venom phenotype. However, these observations are not conclusive, as the results were based on pooled venoms, which present high individual variability. Here we tested the hypothesis of plasticity by influence of confinement and single diet type in the venom composition of 13 adult specimens of Bothrops atrox snakes, maintained under captivity for more than three years. Individual variability in venom composition was observed in samples extracted just after the capture of the snakes. However, composition was conserved in venoms periodically extracted from nine specimens, which presented low variability restricted to the less abundant components. In a second group, composed of four snakes, drastic changes were observed in the venom samples extracted at different periods, mostly related to snake venom metalloproteinases (SVMPs), the core function toxins of B. atrox venom, which occurred approximately between 400 and 500 days in captivity. These data show plasticity in the venom phenotype during the lifetime of adult snakes maintained under captive conditions. Causes or functional consequences involved in the phenotype modification require further investigations.

A etnociência investiga a origem das atitudes e relações humanas com o ambiente. Nesse contexto, a etnozoologia estuda o conhecimento tradicional do homem sobre os animais. O presente trabalho tem como objetivo obter informações sobre o conhecimento tradicional dos moradores da comunidade de Paracari (Monte Alegre, Pará, Brasil) acerca das serpentes, questionando sobre como eles denominam, caracterizam e lidam com esses animais e com os possíveis acidentes. Para isso, durante três campanhas efetuadas nos meses de novembro e dezembro de 2015, questionários contendo tópicos previamente selecionados foram aplicados a 42 moradores selecionados aleatoriamente e que se dispuseram a contribuir. Destes, 95,23% alegaram já terem tido contato com serpentes e, com relação à hostilidade, 59,53% afirmaram matar a serpente independente da espécie. Quanto à ocorrência de acidentes ofídicos no ciclo familiar, 61,90% dos entrevistados confirmaram tal ocorrência, mas apenas 38,46% procuraram atendimento médico. O tratamento de origem popular mais comumente mencionado no caso de picada de cobra foi um extrato de planta artesanal chamado "Pau-X", fabricado, engarrafado e vendido livremente na região. Apenas Eunectes murinus, Lachesis muta e Crotalus durissus foram corretamente identificadas e caracterizadas pelos moradores. Algumas espécies foram identificadas corretamente, mas, em várias ocasiões, foram caracterizadas incorretamente como venenosas ou não. Todos os entrevistados confirmaram nunca ter tido acesso a informações ou participado de práticas educativo-ambientais sobre serpentes, o que pode contribuir diretamente para o uso de práticas inadequadas na identificação das espécies e no tratamento de acidentes ofídicos.

A better knowledge of the burden and risk factors associated with severity due to spider bites would lead to improved management with a reduction of sequelae usually seen for this neglected health problem, and would ensure proper use of antivenoms in remote localities in the Brazilian Amazon. The aim of this study was to analyze the profile of spider bites reported in the state of Amazonas in the Western Brazilian Amazon, and to investigate potential risk factors associated with severity of envenomation. Methodology/Principal Findings We used a case-control study in order to identify factors associated with spider bite severity in the Western Brazilian Amazon from 2007 to 2014. Patients evolving to any severity criteria were considered cases and those with non-severe bites were included in the control group. All variables were retrieved from the official Brazilian reporting systems. Socioeconomical and environmental components were also included in a multivariable analysis in order to identify ecological determinants of incidence and severity. A total of 1,181 spider bites were recorded, resulting in an incidence of 4 cases per 100,000 person/year. Most of the spider bites occurred in males (65.8%). Bites mostly occurred in rural areas (59.5%). The most affected age group was between 16 and 45 years old (50.9%). A proportion of 39.7% of the bites were related to work activities. Antivenom was prescribed to 39% of the patients. Envenomings recorded from urban areas [Odds ratio (OR) = 0.40 (95%CI = 0.30–0.71; pmedian [OR = 0.64 (95%CI = 0.39–0.75; p300 km away from the state capital Manaus [OR = 1.90 (95%CI = 1.28–2.40; p300 km away from the state capital Manaus [OR = 1.53 (95%CI = 1.15–2.02; p = 0.003)] and living in a municipality with a MHSPI 300 km away from the state capital Manaus could be contributing factors to higher severity of spider envenomings in this area, as well as to antivenom underdosage

Comparisons between venoms from snakes kept under captivity or collected at the natural environment are of fundamental importance in order to obtain effective antivenoms to treat human victims of snakebites. In this study, we compared composition and biological activities of Bothrops atrox venom from snakes collected at Tapajós National Forest (Pará State, Brazil) or maintained for more than 10 years under captivity at Instituto Butantan herpetarium after have been collected mostly at Maranhão State, Brazil. Venoms from captive or wild snakes were similar except for small quantitative differences detected in peaks correspondent to phospholipases A2 (PLA2), snake venom metalloproteinases (SVMP) class PI and serine proteinases (SVSP), which did not correlate with fibrinolytic and coagulant activities (induced by PI-SVMPs and SVSPs). In both pools, the major toxic component corresponded to PIII-SVMPs, which were isolated and characterized. The characterization by mass spectrometry of both samples identified peptides that matched with a single PIII-SVMP cDNA characterized by transcriptomics, named Batroxrhagin. Sequence alignments show a strong similarity between Batroxrhagin and Jararhagin (96%). Batroxrhagin samples isolated from venoms of wild or captive snakes were not pro-coagulant, but inhibited collagen-induced platelet-aggregation, and induced hemorrhage and fibrin lysis with similar doses. Results suggest that in spite of environmental differences, venom variability was detected only among the less abundant components. In opposition, the most abundant toxin, which is a PIII-SVMP related to the key effects of the venom, is structurally conserved in the venoms. This observation is relevant for explaining the efficacy of antivenoms produced with venoms from captive snakes in human accidents inflicted at distinct natural environments.

Elucidating the molecular mechanisms underlying snake venom variability provides important clues for understanding how the biological functions of this powerful toxic arsenal evolve. We analyzed in detail individual transcripts and venom protein isoforms produced by five specimens of a venomous snake (Bothrops atrox) from two nearby but genetically distinct populations from the Brazilian Amazon rainforest which show functional similarities in venom properties. Individual variation was observed among the venoms of these specimens, but the overall abundance of each general toxin family was conserved both in transcript and in venom protein levels. However, when expression of independent paralogues was analyzed, remarkable differences were observed within and among each toxin group, both between individuals and between populations. Transcripts for functionally essential venom proteins (“core function” proteins) were highly expressed in all specimens and showed similar transcription/translation rates. In contrast, other paralogues (“adaptive” proteins) showed lower expression levels and the toxins they coded for varied among different individuals. These results provide support for the inferences that (a) expression and translational differences play a greater role in defining adaptive variation in venom phenotypes than does sequence variation in protein coding genes and (b) convergent adaptive venom phenotypes can be generated through different molecular mechanisms. Significance Analysis of individual transcripts and venom protein isoforms produced by specimens of a venomous snake (Bothrops atrox), from the Brazilian Amazon rainforest, revealed that transcriptional and translational mechanisms contribute to venom phenotypic variation. Our finding of evidence for high expression of toxin proteins with conserved function supports the hypothesis that the venom phenotype consists of two kinds of proteins: conserved “core function” proteins that provide essential functional activities with broader relevance and less conserved “adaptive” proteins that vary in expression and may permit customization of protein function. These observations allowed us to suggest that genetic mechanisms controlling venom variability are not restricted to selection of gene copies or mutations in structural genes but also to selection of the mechanisms controlling gene expression, contributing to the plasticity of this important phenotype for venomous snakes.

Background Except for the northern region, where the Amazonian black scorpion, T. obscurus, represents the predominant and most medically relevant scorpion species, Tityus serrulatus, the Brazilian yellow scorpion, is widely distributed throughout Brazil, causing most envenoming and fatalities due to scorpion sting. In order to evaluate and compare the diversity of venom components of Tityus obscurus and T. serrulatus, we performed a transcriptomic investigation of the telsons (venom glands) corroborated by a shotgun proteomic analysis of the venom from the two species. Results The putative venom components represented 11.4% and 16.7% of the total gene expression for T. obscurus and T. serrulatus, respectively. Transcriptome and proteome data revealed high abundance of metalloproteinases sequences followed by sodium and potassium channel toxins, making the toxin core of the venom. The phylogenetic analysis of metalloproteinases from T. obscurus and T. serrulatus suggested an intraspecific gene expansion, as we previously observed for T. bahiensis, indicating that this enzyme may be under evolutionary pressure for diversification. We also identified several putative venom components such as anionic peptides, antimicrobial peptides, bradykinin-potentiating peptide, cysteine rich protein, serine proteinases, cathepsins, angiotensin-converting enzyme, endothelin-converting enzyme and chymotrypsin like protein, proteinases inhibitors, phospholipases and hyaluronidases. Conclusion The present work shows that the venom composition of these two allopatric species of Tityus are considerably similar in terms of the major classes of proteins produced and secreted, although their individual toxin sequences are considerably divergent. These differences at amino acid level may reflect in different epitopes for the same protein classes in each species, explaining the basis for the poor recognition of T. obscurus venom by the antiserum raised against other species.

Made available in DSpace on 2018-12-11T17:14:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-06 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Many scorpion accidents occur in the Brazilian Amazonian region and are frequently caused by Tityus obscurus. Approximately 5% of the crude venom of this species is composed of short linear, non-disulfide-bridged peptides, which have not been intensively investigated. As a consequence, only a few of these peptides have been structurally and functionally characterized to date. In the present paper, the peptide fraction of the venom was subjected to peptide profiling using an LCMS-IT-TOF/MS and MSn system. The analysis detected 320 non-disulfide bond-containing peptides (NDBPs), of which twenty-seven had their sequences assigned; among them, thirteen peptides were characterized, constituting novel toxins in T. obscurus venom. Some of the novel peptides showed similarities to hypotensin-like toxins, while other peptides appear to be natural fragments of neurotoxins. The novel peptides were submitted to a series of bioassays, revealing that many are multifunctional toxins that cause, for example, pain, edema formation and hemolysis to potentiate strong inflammatory processes and alterations in the locomotion and lifting activities in the victims of stinging. Knowledge of the complex matrix of peptides composing the venom of T. obscurus will contribute to better understanding of the complex mechanism of envenoming caused by stinging accidents. Significance The scorpion Tityus obscurus causes many envenoming accidents of medical importance in Brazilian Amazon region; despite to this, very few is known about the toxinology of this animal. The knowledge about the venom composition and mechanisms of action is very important to understand the physiopathology processes related to the envenoming caused by this animal. The proteopeptidomic investigations of scorpion venoms in general have focused mainly the neurotoxins (which are disulfide bonds containing peptides) and large proteins. The short, linear, non-disulfide bonds containing peptides (NDBPs) represent up to 5% of scorpion venom compositions; however, they have been few investigated in comparison with the neurotoxins. The present study used a mass spectrometric approach to detect 320 NDBPs and to sequence 27 of them; pharmacological assays permitted to characterize 13 NDBPs as novel toxins involved with inflammation, pain and edema formation. Dept. Biology/CEIS Institute of Biosciences of Rio Claro University of São Paulo State (UNESP) Laboratory of Biological Research Amazon College/Amazon University UNAMA Laboratory of Pharmacology Butantan Institute Dept. Biology/CEIS Institute of Biosciences of Rio Claro University of São Paulo State (UNESP) FAPESP: 2011/51684-1

Bothrops atrox snakes are the leading cause of snake bites in Northern Brazil. The venom of this snake is not included in the antigen pool used to obtain the Bothrops antivenom. There are discrepancies in reports on the effectiveness of this antivenom to treat victims bitten by B. atrox snakes. However, these studies were performed using a pre-incubation of the venom with the antivenom and, thus, did not simulate a true case of envenomation treatment. In addition, the local lesions induced by Bothrops venoms are not well resolved by antivenom therapy. Here, we investigated the efficacy of the Bothrops antivenom in treating the signs and symptoms caused by B. atrox venom in mice and evaluated whether the combination of dexamethasone and antivenom therapy enhanced the healing of local lesions induced by this envenomation. In animals that were administered the antivenom 10 minutes after the envenomation, we observed an important reduction of edema, dermonecrosis, and myonecrosis. When the antivenom was given 45 minutes after the envenomation, the edema and myonecrosis were reduced, and the fibrinogen levels and platelet counts were restored. The groups treated with the combination of antivenom and dexamethasone had an enhanced decrease in edema and a faster recovery of the damaged skeletal muscle. Our results show that Bothrops antivenom effectively treats the envenomation caused by Bothrops atrox and that the use of dexamethasone as an adjunct to the antivenom therapy could be useful to improve the treatment of local symptoms observed in envenomation caused by Bothrops snakes., Author summary Bothrops atrox is the dominant species responsible for accidental human snake bites in Northern Brazil. The efficacy of antivenom therapy to correct the systemic disturbances, including hemostatic disorders, caused by Brazilian Bothrops is well known. However, two fundamental issues need to be addressed in this region. (1) There are concerns regarding the effectiveness of the antivenom to treat Bothrops snake bites in this region since Bothrops atrox venom is not used as an antigen to obtain the Bothrops antivenom in Brazil, and (2) the efficacy of the antivenom therapy in reversing local injuries induced by Bothrops venoms is low. Thus, our study aimed to assess the effectiveness of antivenom therapy alone or in combination with dexamethasone to treat experimental envenomation induced by Bothrops atrox venom in mice. Our results showed that the Brazilian Bothrops antivenom effectively reversed the systemic disturbances caused by this envenomation and combining the antivenom therapy with dexamethasone accelerated the regression of inflammatory edema and the regeneration of skeletal muscle that was damaged by the venom.

Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p0.05) and significantly reduced (p0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd was administered in combination with BA. The SDS-PAGE profiles showed that several of the BaV protein (SDS-PAGE) and enzyme (zymography) bands were not detected when the venom was pre-incubated, and Western blot revealed that this was not caused by the AEBd enzymes observed in the zymogram. The "pseudo inhibition" observed after pre-incubation in this study may be due to the presence of tannins in the extract, which could act as chelating agents, removing metalloproteins and Ca²⁺ ions and thus inhibiting hemorrhagin and PLA₂ activity. However, when administered according to traditional methods, B. dichotoma extract was effective in blocking BaV-induced edematogenic activity and had an additional effect on inhibition of this activity by BA.

This study analyzes the effectiveness of extracts from five plant species used for treating venomous snakebites in the region of Santarem, in central Brazilian Amazonia. The aqueous extracts of Bellucia dichotoma Cogn., Connarus favosus Planch., Aniba fragrans Ducke,Plathymenia reticulata Benth., and Philodendron megalophyllum Schott were prepared by decoction at 70°C and lyophilized. Inhibition of the enzymatic activity and biological effects of the crude venom of Bothrops atrox was tested by pre-incubating different concentrations (1:1 to 1:30 venom:extract, w/w) for 30 min at 37°C. These results confirm traditional local knowledge on the anti-toxin powers of the native plants of the Amazon basin and the potential of these extracts as toxin inhibitors and as an alternative or complement for the standard treatment of snakebite based on the application of antisera. Key words: Natural inhibitors, Bothrops atrox, Bellucia dichotoma, Connarus favosus, Plathymenia reticulata, Philodendron megalophyllum, Aniba fragrans.

There are a great number of studies about Brazilian scorpions. However, little is known about the venom of scorpions of northern Brazil, mainly about Tityus obscurus, which is responsible for the most number of accidents in the Amazon. Thus, this study aimed to evaluate some pharmacological effects of T. obscurus venom in rats and mice. In rats, the venom (10 mg/kg i.p.) caused hemorrhagic patches in the lung parenchyma but did not lead to pulmonary edema. There was a decrease in general activity, observed in the activity box after venom injection. The venom did not induce changes in the occurrence and intensity of experimentally induced convulsions, nor did it cause hippocampal neuronal loss. In mice, the LD50 obtained was 3.13 mg/kg (i.p.). Different doses of the venom (0.2; 1; 5; 10; 15 μg/30 μL per hind paw) induced edematogenic and moderate nociceptive activity in mice. The Tiyus serrulatus venom used as comparison caused more intense symptomatology in mice. Comparing to the venom of other Tityus scorpions of medical importance, that have convulsant and intense nociceptive effects and cause lung edema, as described in the literature, we can conclude that the venom of T. obscurus probably has different characteristics.

Lancehead pit-vipers (Bothrops genus) are an extremely diverse and medically important group responsible for the greatest number of snakebite envenomations and deaths in South America. Bothrops atrox (common lancehead), responsible for majority of snakebites and related deaths within the Brazilian Amazon, is a highly adaptable and widely distributed species, whose venom variability has been related to several factors, including geographical distribution and habitat type. This study examined venoms from four B. atrox populations (Belterra and Santar&eacute, m, PA, Pres. Figueiredo, AM and S&atilde, o Bento, MA), and two additional Bothrops species (B. jararaca and B. neuwiedi) from Southeastern region for their coagulotoxic effects upon different plasmas (human, amphibian, and avian). The results revealed inter&ndash, and intraspecific variations in coagulotoxicity, including distinct activities between the three plasmas, with variations in the latter two linked to ecological niche occupied by the snakes. Also examined were the correlated biochemical mechanisms of venom action. Significant variation in the relative reliance upon the cofactors calcium and phospholipid were revealed, and the relative dependency did not significantly correlate with potency. Relative levels of Factor X or prothrombin activating toxins correlated with prey type and prey escape potential. The antivenom was shown to perform better in neutralising prothrombin activation activity than neutralising Factor X activation activity. Thus, the data reveal new information regarding the evolutionary selection pressures shaping snake venom evolution, while also having significant implications for the treatment of the envenomed patient. These results are, therefore, an intersection between evolutionary biology and clinical medicine.

Boids are primitive snakes from a basal lineage that is widely distributed in Neotropical region. Many of these species are both morphologically and biogeographically divergent, and the relationship among some species remains uncertain even with evolutionary and phylogenetic studies being proposed for the group. For a better understanding of the evolutionary relationship between these snakes, we cytogenetically analysed 7 species and 3 subspecies of Neotropical snakes from the Boidae family using different chromosomal markers. The karyotypes of Boa constrictor occidentalis, Corallus hortulanus, Eunectes notaeus, Epicrates cenchria and Epicrates assisi are presented here for the first time with the redescriptions of the karyotypes of Boa constrictor constrictor, B. c. amarali, Eunectes murinus and Epicrates crassus. The three subspecies of Boa, two species of Eunectes and three species of Epicrates exhibit 2n = 36 chromosomes. In contrast, C. hortulanus presented a totally different karyotype composition for the Boidae family, showing 2n = 40 chromosomes with a greater number of macrochromosomes. Furthermore, chromosomal mapping of telomeric sequences revealed the presence of interstitial telomeric sites (ITSs) on many chromosomes in addition to the terminal markings on all chromosomes of all taxa analysed, with the exception of E. notaeus. Thus, we demonstrate that the karyotypes of these snakes are not as highly conserved as previously thought. Moreover, we provide an overview of the current cytotaxonomy of the group. © 2016 Viana et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

We present here a new record for Amphisbaena michelli for the western of Pará State, northern Brazil. The specimen was found in the municipality of Belterra, on the right bank of the Tapajós River, and represents an expansion ca. 350 km west of the previously known geographic distribution of the species.

In Latin America, Bothrops snakes account for most snake bites in humans, and the recommended treatment is administration of multispecific Bothrops antivenom (SAB – soro antibotrópico). However, Bothrops snakes are very diverse with regard to their venom composition, which raises the issue of which venoms should be used as immunizing antigens for the production of pan-specific Bothrops antivenoms. In this study, we simultaneously compared the composition and reactivity with SAB of venoms collected from six species of snakes, distributed in pairs from three distinct phylogenetic clades: Bothrops, Bothropoides and Rhinocerophis. We also evaluated the neutralization of Bothrops atrox venom, which is the species responsible for most snake bites in the Amazon region, but not included in the immunization antigen mixture used to produce SAB. Using mass spectrometric and chromatographic approaches, we observed a lack of similarity in protein composition between the venoms from closely related snakes and a high similarity between the venoms of phylogenetically more distant snakes, suggesting little connection between taxonomic position and venom composition. P-III snake venom metalloproteinases (SVMPs) are the most antigenic toxins in the venoms of snakes from the Bothrops complex, whereas class P-I SVMPs, snake venom serine proteinases and phospholipases A2 reacted with antibodies in lower levels. Low molecular size toxins, such as disintegrins and bradykinin-potentiating peptides, were poorly antigenic. Toxins from the same protein family showed antigenic cross-reactivity among venoms from different species; SAB was efficient in neutralizing the B. atrox venom major toxins. Thus, we suggest that it is possible to obtain pan-specific effective antivenoms for Bothrops envenomations through immunization with venoms from only a few species of snakes, if these venoms contain protein classes that are representative of all species to which the antivenom is targeted., Author Summary Snakebite envenomation is a serious health issue in Latin America, particularly in the Amazon, where antivenom administration may be delayed due to logistic constraints. Bothrops snakes are involved in most of the snakebite-related accidents in Brazil. This work reports a comparative study of the toxin composition and antigenicity of the Bothrops venoms used to prepare the commercial antivenom and its effectiveness against the venom from Bothrops atrox, a prevalent Amazon species that is not included in the pool. Our data show a lack of connection between Bothrops taxonomic identity and venom composition. We also show that different toxins display distinct reactivity with the tested antivenom. However, the antivenom reacted similarly with each class of toxin present in the venoms of the different snakes studied. Important evidence was the neutralization of the major toxic effects of B. atrox venom, not included in the mixture of antigens used to produce the antivenom. Based on the observed antigenicity of the distinct protein classes of toxins, we suggest that it is possible to obtain pan-specific and efficient Bothrops antivenoms via immunization with venoms from a few species of snakes that are representative of the protein composition of a large number of targeted species.