Mycoplasma spp. in captive snakes (Boa constrictor and Bothrops atrox) from Brazil / Mycoplasma spp. em cobras de cativeiro (Boa constrictor e Bothrops atrox) do Brasil

ABSTRACT: Although rare, mycoplasmas are included among the causes of respiratory diseases in reptiles and, in the order Squamata, three reports of these microorganisms causing diseases in pythons have already been reported. This study aimed to evaluate the occurrence of Mycoplasma species in captive snakes. A total of 26 snakes of the families Pythonidae (13), Boidae (7), Viperidae (5) and Colubridae (1) from RioZoo, Brazil, were evaluated. Animals were examined to determine clinical signs consistent with any infectious disease. Tracheal swab samples from snakes were collected in Frey medium and analyzed for the presence of Mycoplasma spp.by isolation and a genus-specific PCR. DNA sequencing analyses of six positive samples by PCR were carried out to identify the species. Using isolation 19.23% (5/26) was positive, while 65.38% (17/26) of the animals were positive by PCR. Based on the analyses of the six sequences obtained, there was similarity with a Mycoplasma spp. previously described in a phyton and, M. agassizii and M. testudineum reported in chelonians. This is the first report of Mycoplasma spp. in animals of the families Boidae and Viperidae. Mycoplasma spp. were detected in snakes with and without clinical signs. The mycoplasmas reported resented identity (range, 95% to 100%) to others already described in reptiles. There was no relationship between the presence of Mycoplasma spp. and clinical signs.

RESUMO: Embora raros, os micoplasmas estão incluídos entre as causas de doenças respiratórias em répteis e, na ordem Squamata, já foram realizados três relatos destes microrganismos causando doença em pítons. Este estudo teve como objetivo avaliar a ocorrência de espécies de Mycoplasma em serpentes em cativeiro. Foram avaliadas 26 serpentes das famílias Pythonidae (13), Boidae (7), Viperidae (5) e Colubridae (1) do RioZoo, Brasil. Os animais foram examinados para determinar sinais clínicos consistentes com qualquer doença infecciosa. Amostras de swab traqueal de cobras foram coletadas em meio Frey e analisadas por isolamento microbiológico e pela técnica da PCR para identificar Mycoplasma spp. As amostras positivas para o gênero Mycoplasma spp. foram submetidas ao sequenciamento genético para identificação das espécies. No isolamento, 19,23% (5/26) foram positivos, enquanto 65,38% (17/26) dos animais foram positivos por PCR. Com base nas análises das seis sequências obtidas, houve similaridade com o Mycoplasma spp. descrito anteriormente em um píton e M. agassizii e M. testudineum encontrados em quelônios. Este é o primeiro relato de Mycoplasma spp. em animais das famílias Boidae e Viperidae. Mycoplasma spp. foi detectado em serpentes com e sem sinais clínicos. Os micoplasmas encontrados apresentaram semelhança genética com outros já descritos em répteis. Não houve relação entre a presença de Mycoplasma spp. e sinais clínicos.

Hemostatic evaluation of rabbits envenomed with Bothrops alternatus treated with anti-bothropic serum, desmopressin and tranexamin acid / Avaliação hemostática de coelhos envenenados com Bothrops alternatus e tratados com soro antibotrópico, desmopressina e ácido tranexâmico

In Brazil, snakes from the Bothrops genus are responsible for thousands of accidents, and their venoms are mainly composed of proteolytic enzymes. Although the antibothropic serum produced by the Brazilian Institutes is remarkably efficient, more studies are necessary, especially in veterinary medicine. The venom contain enzymes and non-enzymatic proteins that interfere with hemostasis leading to hemorrhage or even thrombosis. Possible treatment associations with known bothropic antivenom were the reason for the development of the present study. The aim of this study was to evaluate hemostasis alterations caused by Bothrops alternatus venom in rabbits followed by treatments with anti-bothropic serum, tranexamic acid and desmopressin. Twenty New Zealand rabbits were distributed into five groups (n=4) that were experimentally envenomed with 150mcg/kg of B. alternatus venom via intramuscular injection and treated as follow: Group 1 (G1) was the positive control and received venom and PBS/BSA; Group 2 (G2) was treated with tranexamic acid; Group 3 (G3) with desmopressin; Group 4 (G4) with tranexamic acid and anti-bothropic serum; and Group 5 (G5) with anti-bothropic serum and desmopressin. Blood samples were collected before venom administration, and one, four, eight and 12 hours after, for Partial activated partial thromboplastin time, Prothrombin Time, Thrombin Time and fibrinogen evaluation. Thrombin generation (TG) test was carried out with a pool of samples from final times (8 and 12h). At the end of 12h, all animals were euthanized and necropsy was conducted. Samples from muscle tissue, heart, lungs and kidney were analyzed. Classic coagulation tests showed no significant differences amongst groups and times. However, TG indicated that the venom causes a hypocoagulability state, which was not reversed by proposed treatments. Histology showed muscle inflammation, hemorrhage and necrosis, as well as hemorrhage in other tissues with no differences amongst groups. B. alternatus envenomation causes hypocoagulability detected by TG assay, but not through classical coagulation tests. The use of tranexamic acid and desmopressin for hemostasis stabilization after inoculation of the venom did not show advantage in coagulation restoration.(AU)

No Brasil, as serpentes do gênero Bothrops são responsáveis por milhares de acidentes, e seus venenos são compostos principalmente de enzimas proteolíticas. Embora o soro antiofídico produzido pelos institutos brasileiros seja notavelmente eficiente, mais estudos são necessários, especialmente na medicina veterinária. O veneno contem enzimas e proteínas não-enzimáticas que interferem com a hemostasia levando a hemorragias ou trombose. A associação de outros tratamentos ao soro antibotrópico foi a razão para o desenvolvimento do presente estudo. O objetivo deste estudo foi avaliar as alterações da hemostasia causadas pelo veneno de Bothrops alternatus em coelhos, após tratamento com soro antibotrópico, ácido tranexâmico e desmopressina. Vinte coelhos da Nova Zelândia foram distribuídos em cinco grupos (n = 4) que foram submetidos a experimentos com 150mcg/kg de veneno de B. alternatus por injeção intramuscular. O Grupo 1 (G1) foi o controle positivo e recebeu veneno e PBS / BSA, enquanto o Grupo 2 (G2) foi tratado com ácido tranexâmico, o Grupo 3 (G3) com desmopressina, o Grupo 4 (G4) com ácido tranexâmico e soro antibotrópico, e o Grupo 5 (G5) com soro antibotrópico e desmopressina. As amostras de sangue foram coletadas antes da administração do veneno, e uma, quatro, oito e 12 horas após os tratamentos para realização de tempo de tromboplastina parcial ativada parcial (TTPa), tempo de protrombina (TP), tempo de trombina (TT) e mensuração de fibrinogênio. Para o ensaio de geração de trombina (TG) foi realizado com um pool de amostras nos tempos finais (8 e 12h). Ao final das 12h, todos os animais foram sacrificados e a necropsia foi realizada. Amostras de tecido muscular, coração, pulmões e rins foram analisadas. Os testes TTPa, TP, TT e fibrinogênio não mostraram diferenças significativas entre os grupos e os tempos. No entanto, o TG indicou que o veneno causa um estado de hipocoagulabilidade, que não foi revertido pelos tratamentos propostos. Na histologia, foram observadas inflamação muscular, hemorragia e necrose, além de hemorragia em outros tecidos, sem diferenças entre os grupos. O envenenamento por B. alternatus causa hipocoagulabilidade detectada mais precocemente pelo teste de geração de trombina. O uso de ácido tranexâmico e desmopressina para estabilização da hemostasia após a inoculação do veneno não mostrou vantagem na restauração da coagulação.(AU)

Clinical Profile of Snake Envenomation with Complications and Outcome in a Tertiary Health Care Centre, Kalburgi.

Bites by snakes represent an important health problem in the tropical world including India. The true incidence of snakebites is difficult to assess and often is under reported. Though the exact number of snake bites is unknown, an estimated 5.4 million people are bitten each year with up to 2.7 million envenoming. Around 81,000 to 1,38,000 people die each year because of snake bites, and around three times as many amputations and other permanent disabilities are caused by snakebites annually. We wanted to determine frequency of various modes of presentation, complications and management of snakebite.METHODSAll patients presenting to emergency department of Basaveshwara Teaching and General Hospital with history of snakebite and features of envenomation were included in the study. All these patients were given Anti-Snake Venom. Symptomatic treatment of complications was done. Semi-structured questionnaire was filled during hospital stay to analyse various characteristics of snakebite and people’s attitude towards this problem.RESULTS75 cases of snakebite admitted to Basaveshwara Teaching & General Hospital, Gulbarga between January 2018 to November 2019 were studied. Males [50 (62.5%)] were affected more than females [25 (31.5%)] in the ratio of 2:1. Most common age group is 20-40 years [45 (60%)]. Snakebite is a common health hazard in rural areas [55 (73.34%)]. Agricultural labourers (68.6%) are the major sufferers with majority of bites occurring outdoor 60 (80%). 71 (94.66%) bites occurred over limbs, out of which 38 (50.66%) were in lower limbs. Most bites occur between 12 Noon to 12 Midnight 60 patients (81.2%). Out of 75 patients, 60 (80%) were poisonous snakebites, 20 (20%) were non-poisonous snakebites. Among 60 (80%) poisonous bites, 32 (53.34%) had neurotoxic manifestations and 28 (46.66%) had hemotoxic manifestations.CONCLUSIONSSnakebite is common in rural setup, in agricultural labourers. Most common victims are males in the age group between 20-40 years. Poisonous bites were more common than non-poisonous snake bites in this study. In neurotoxic envenomation ptosis was the commonest and earliest symptom while in hemotoxic envenomation, most common symptoms were bleeding from bite site and hematuria. The commonest complications were acute renal failure and respiratory paralysis. Shock, infection and gangrene were also noted in some patients. Maximum mortality was observed in patients who were admitted after 24 hours. Early hospitalization and timely ASV administration were the corner stones in the treatment of snakebite.

Hemorrhagic stroke following viper bites and delayed antivenom administration: three case reports from the Western Brazilian Amazon

Abstract Snakebites were included by the World Health Organization in their list of neglected diseases. In Latin America, most snakebites are caused by species of the Viperidae family, notably by the genus Bothrops. Bothrops atrox accounts for 90% of the cases of envenoming in the Brazilian Amazon. In this report, we present a series of three cases of snakebites that evolved with hemorrhagic stroke due to delays in the access to antivenom in the Brazilian Amazon, being fundamental for diagnosis to validate the clinical suspicion and make decisions that would improve the treatment and prognosis of the patients.

Epidemiological study of snakebite cases in Brazilian Western Amazonia

Abstract INTRODUCTION Brazil has the largest number of snakebite cases in South America, of which the large majority is concentrated in the Midwest and North. METHODS In this descriptive observational study, we assessed the epidemiological and clinical snakebite cases referred to the Centro de Medicina Tropical de Rondônia from September 2008 to September 2010. RESULTS We followed up 92 cases from admission until discharge, namely 81 (88%) men and 11 (12%) women, with a mean age of 37 years, and mainly from rural areas (91.3%). The snakebites occurred while performing work activities (63%) during the Amazon rainy season (78.3%). The vast majority of individuals presented from the Porto Velho microregion (84.7%). Approximately 95.6% of the snakebites were caused by snakes of the genus Bothrops, followed by two lachetics and two elapidics cases. Surgery was performed in 10 cases (9 fasciotomies in the lower limb and 1 amputation). No deaths were reported in this study, but 4 cases (4.3%) developed sequelae in the lower limb. CONCLUSIONS This study can contribute to a better understanding of envenomation in the state of Rondônia and thus can be useful for identifying real conditions that can increase the incidence of snakebites in this region. Moreover, the study results can serve as a basis for improving educational campaigns designed to prevent these types of snakebites, as well as for preserving snakes.

Could mesenchymal stem cell therapy help in the treatment of muscle damage caused by Bothrops alternatus venom? / Podem as células tronco mesenquimais auxiliarem no tratamento da lesão muscular causada pelo veneno de Bothrops alternatus?

ABSTRACT: The aim of this study was to evaluate the use of mesenchymal stem cells (MSC) in the treatment of myonecrosis induced by Bothrops alternatus venom in rats. Seventy-five male adult Wistar rats were divided into three experimental groups. G1 and G2 were injected in the gastrocnemius muscle with 120μg of B. alternatus venom, while G3 received 200μL of PBS only. Three days after the venom injection, 12 rats from G1 were treated with 5.0 x 106 MSC in PBS, whereas G2 and G3 rats received PBS. Every three days, blood and muscle samples of five animals from each group were taken for serum biochemical and pathological analyses. Histological examinations showed more intense muscle lesions following MSC treatment, characterized by disorganization and loss of muscle fibers, with focal necrosis and inflammatory infiltration by mononuclear cells. In conclusion, the use of MSC for the treatment of local damage caused by inoculation of B. alternatus venom impaired muscle regeneration and interfered in the healing process.

RESUMO: O objetivo do presente trabalho foi avaliar a utilização das células tronco mesenquimais (MSC) no tratamento da mionecrose induzida pelo veneno de Bothrops alternatus em ratos. 75 ratos Wistar adultos foram distribuídos em três grupos experimentais. G1 e G2 receberam 120μg de veneno de B. alternatus, enquanto o G3 recebeu apenas 200μL de PBS. Três dias após a administração do veneno, os ratos do grupo G1 foram tratados com 5.0 x 106 MSC, enquanto G2 e G3 receberam exclusivamente PBS. A cada três dias, amostras de sangue e tecido muscular foram coletadas de cinco animais de cada grupo para avaliação bioquímica sérica e patológica, respectivamente. A análise histológica revelou lesão muscular mais intensa após a aplicação das MSC, caracterizada pela desorganização e perdas das fibras musculares, com necrose focal e infiltrado inflamatório mononuclear. É possível concluir que a utilização das MSC para o tratamento da lesão local causada pela inoculação do veneno B. alternatus deteriorou a regeneração muscular e interferiu com o processo de cicatrização.

A study of snakebite envenomation cases admitted to egyptian national poisoning center

Introduction: Snakebite is an Egyptian health problem since ancient Egypt. Meanwhile, there is still no controlled geographical and medical studies on locally prevalent snake family intoxication. Aim and Methods: The present study aimed to investigate local snakebites presentations, management, prognosis, and the effect of the polyvalent antivenom, locally produced by the Holding Company for Biological Products and Vaccines (VACSERA) through the year 2015. Results: A total of 87 snakebites caused by venomous and nonvenomous species were recorded. Most cases were middle-aged males. Most cases presented in summer and in the evening time. Two major groups of venomous Egyptian snakes were identified, Viperidae and Elapidae species, based on history, characteristic symptoms, and laboratory findings. Most snakebites (56 cases) were reported to be nonvenomous bites (64.4%). Twenty-one cases (24.1%) of snakebites were reported to be venomous bites by Elapidae snakes and 10 cases (11.5%) were reported to be venomous bites by Viperidae snakes. Antivenom was administered before referral to 37 (42.5%) of cases, and 19 only of them were victims of venomous snakes. Thirty-six (41.3%) patients received antivenom during admission including all cases of the venomous bites and 8.9% of nonvenomous bites' cases. Conclusion: Patients who had moderate or severe symptoms were effectively treated with VACSERA's polyvalent antivenom, with doses related to the severity grading and snake species identification. Additional antivenom doses were repeated on the bases of the clinical condition. Many snakebite victims referred from primary health centers received inadequate or nonnecessary doses of antivenom. No cases of anaphylaxis were recorded. There were no mortalities with current National Center for Clinical and Environmental Toxicology's protocol of treatment.

Russell’s viper venom induced nephrotoxicity, myotoxicity, and hepatotoxicity—Neutralization with gold nanoparticle conjugated 2-hydroxy-4-methoxy benzoic acid in vivo.

Snakebite is one of the major neglected tropical diseases and health hazard that leads to significant mortality, particularly in rural populations of tropical and subtropical countries including India. Antisnake venom serum (ASVS) is the only specific treatment against snake envenomation. Available treatment i.e. ASVS have many limitations not only low efficiency but also considerable side effects. Search for alternative ASVS is a major domain in toxinology research. Targeted drug therapy using nanoparticles, an emerging area of nanotechnology, is one such alternative. Here, we studied neutralization of ing Russell’s viper venom (RVV) induced toxicity (nephrotoxicity, myotoxicity, hepatotoxicity) with gold nanoparticle-conjugated 2-hydroxy-4-methoxy benzoic acid (GNP-HMBA) in male albino mice. We conjugated 2-hydroxy-4-methoxy benzoic acid (HMBA) with gold nanoparticle (GNP) by adsorption method, and physico-chemical characterization were done by DLS, ZETA potential, FTIR and TEM. Swiss male albino mice were divided into four groups viz., sham control, venom control, HMBA treated and GNP-HMBA treated. Each group had four mice (n=4). RVV was injected in all groups except sham control. Groups 3 and 4 had treatment with HMBA and GNP-HMBA, respectively. After 24 h, blood and urine were collected. Serum LDH, CK, SGPT, SGOT, γ-GT, ACP, ALP, urea, creatinine and urinary calcium and urinary phosphorus were measured. The hydrodynamic diameter of GNP-HMBA was 65-75 nm and TEM diameter was 18-28 nm. The serum/urine parameters were found significantly increased in venom control group. Degree of RVV neutralization was GNP-HMBA > HMBA. Treatment with GNP-HMBA showed partial protection of histopathological changes in RVV-induced kidney and liver tissues. It may be concluded that GNP-HMBA neutralized RVV-induced toxicities (nephrotoxicity, myotoxicity and hepatotoxicity) in male albino mice. Further studies are warranted in the development of alternative herbal-nanoparticle antidote against snake venom induced toxicity.

Characterization of the most promising fraction of Swietenia macrophylla active against myotoxic phospholipases A2: identification of catechin as one of the active compounds / Caracterización de la fracción más promisoria de Swietenia macrophylla activa contra fosfolipasas A2 miotoxicas: identificación de catequina como uno de los compuestos activos

Background: The pharmacological effects produced by snakebite accidents involve the actions of several enzymes, of which those of the phospholipases A2 (PLA2) exhibit a wide variety of effects such as edema and myotoxicity. Some plant extracts have been antagonists of crude snake venoms and toxins. Based on promising bioactivity, Swietenia macrophylla King was selected for further studies. Objective: The purpose of this study was to identify the PLA2 inhibitors present in the crude extract of S. macrophylla that could be promising leads in neutralizing the local effects of ophidian accidents. Methods: Bioassay-guided fractionation of the ethanolic extract of the leaves of S. macrophylla lead to the detection of (+)-catechin, characterized through gas chromatography coupled with mass spectrometry (GC-MS), and confirmed by HPLC. The PLA2 inhibitory activity was measured with the Dole method and a spectrophotometric assay with 4-Nitro-3 octanoyloxy-benzoic acid (4N3OBA). Cytotoxicity was done on C2C12 murine myoblast. Results: Fraction F5 and (+)-Catechin inhibited the PLA2 activity of B. asper venom, in a dosedependent way. In addition, (+) Catechin showed an inhibition level of 83.1 ± 3.1 % of the enzymati activity of one PLA2 purified from the venom of Crotalus durissus cumanensis using 4N3OBA as substrate. Also the ethanolic extract and fraction F5 showed inhibition of the cytotoxicity induced by the Bothrops atrox venom and their Lys 49 PLA2 (80 and 100% respectively). Molecular docking results suggested that OH from 4´ and 5' carbons of (+)-catechin could form hydrogen bonds with carboxylate moiety of residue Asp49, while OH from 5 could form a hydrogen bond with Asn 6. Additional Van der Waals interactions were also proposed. Conclusion: Swietenia macrophylla exhibited strong inhibitory activity against PLA2s enzymes. Catechin, one of the components in the active fraction F5, is proposed as being partially responsible for the bioactivity.

Antecedentes: Los efectos farmacológicos producidos en el accidente ofídico implican la acción de varias enzimas, como las fosfolipasas A2 (PLA2), que exhiben una amplia variedad de efectos como edema y miotoxicidad. Algunos extractos de plantas han demostrado ser antagonistas de los venenos crudos y sus toxinas. En base a una bioactividad promisoria previa, Swietenia macrophylla King fue seleccionada para estudios posteriores. Objetivo: El propósito de este estudio fue identificar metabolitos inhibidores de PLA2 presentes en el extracto crudo de S. macrophylla que podrían ser prometedores en la neutralización de los efectos locales del accidente ofídico. Métodos: Un fraccionamiento biodirigido del extracto etanólico de hojas de S. macrophylla llevó a la detección de la (+)-catequina mediante cromatografía de gases acoplada a espectrometría de masas (GC-MS), y se confirmó mediante HPLC. La actividad inhibidora de PLA2 se determinó por el método de Dole y un ensayo espectrofotométrico con Ácido 4-Nitro-3-octanoiloxibenzoico (4N3OBA). La citotoxicidad se determinó en mioblastos C2C12 murinos. Resultados: La fracción F5 de S. macrophylla, mostró la mayor inhibición del veneno de Bothrops asper y PLA2s aisladas de B. atrox y Crotalus durissus cumanensis e inhibición completa de la citotoxicidad. La (+)-catequina fue el metabolito más abundante en F5, mostró una inhibición de PLA2 de 89,8%, 81,1% y 74,3%, a diferentes relaciones de veneno: catequina. Mediante estudios de docking molecular se demostró la unión de la (+)-catequina al sitio activo de la PLA2. Conclusión: Swietenia macrophylla exhibió una fuerte actividad inhibitoria sobre las enzimas PLA2s. La (+)-catequina, uno de los metabolitos presentes en la fracción activa F5, se propone como uno de los compuestos responsables de la bioactividad.

Caryospora bigenetica (Apicomplexa: Eimeriidae) in South America: new hosts and distribution records / Caryospora bigenetica (Apicomplexa: Eimeriidae) na América do Sul: Registro de novos hospedeiros e distribuição

The coccidian Caryospora bigenetica was first described in the snake Crotalus horridus (Viperidae) from United States of America. This study represents the first record of the occurrence of C. bigenetica in snakes in South America. Feces were sampled between November 2013 and May 2014 from 256 wild snakes maintained in scientific breeding facilities in the states of Mato Grosso do Sul (MS; n = 214) and Rio de Janeiro (RJ; n = 42), Brazil. Caryospora bigenetica was found in 14 (5.6%) snakes, all belonging to the family Viperidae. Ten Bothrops moojeni and two Crotalus durissus from MS were infected. The coccidian was also found in one C. durissus and in one Bothrops jararacussu from the state of RJ. The oocysts were spherical with a double wall, the exterior lightly mammillated, striations apparent in transverse view, 13.0 µm (12 – 14); polar granule fixed in the internal wall. Sporocysts oval or pyriform, 10.0 × 8.0 µm (9 – 11 × 8 – 9); Stieda body discoid; sub-Stieda body present; sporocyst residuum present, formed by a group of spheroid bodies between sporozoites. This study increases the number of viperid hosts of C. bigenetica and expands the geographical distribution to South America.

O coccídio Caryospora bigenetica foi descrito na serpente Crotalus horridus (Viperidae) nos Estados Unidos da América. Este estudo representa o primeiro registro da ocorrência de C. bigenetica em serpentes da América do Sul. Amostras de fezes foram obtidas, entre novembro de 2013 e maio de 2014, de 256 serpentes silvestres mantidas em um criatório científico nos Estados do Mato Grosso do Sul (MS; n = 214) e Rio de Janeiro (RJ; n = 42), Brasil. Caryospora bigenetica foi encontrada em 14 (5,6%) serpentes, todas pertencentes à família Viperidae. Dez Bothrops moojeni e duas Crotalus durissus de MS estavam infectadas. O coccídio também foi encontrado em uma C. durissus e uma Bothrops jararacussu do Estado do Rio de Janeiro. Os oocistos foram esféricos, com parede dupla, sendo a externa ligeiramente mamilonada, estriações aparentes transversalmente, 13.0 µm (12 – 14); grânulo polar junto à parede interna. Esporocisto oval ou piriforme, 10.0 × 8.0 µm (9 – 11 × 8 – 9); corpo de Stieda discóide; sub-Stieda presente; resíduo do esporocisto presente, formado por um grupo de corpos esféricos entre os esporozoítos. Este estudo aumenta o número de hospedeiros viperídeos de C. bigenetica e expande a distribuição geográfica para a América do Sul.

Caryospora bigenetica (Apicomplexa: Eimeriidae) in South America: new hosts and distribution records

The coccidian Caryospora bigenetica was first described in the snake Crotalus horridus (Viperidae) from United States of America. This study represents the first record of the occurrence of C. bigenetica in snakes in South America. Feces were sampled between November 2013 and May 2014 from 256 wild snakes maintained in scientific breeding facilities in the states of Mato Grosso do Sul (MS; n = 214) and Rio de Janeiro (RJ; n = 42), Brazil. Caryospora bigenetica was found in 14 (5.6%) snakes, all belonging to the family Viperidae. Ten Bothrops moojeni and two Crotalus durissus from MS were infected. The coccidian was also found in one C. durissus and in one Bothrops jararacussu from the state of RJ. The oocysts were spherical with a double wall, the exterior lightly mammillated, striations apparent in transverse view, 13.0 µm (12 14); polar granule fixed in the internal wall. Sporocysts oval or pyriform, 10.0 × 8.0 µm (9 11 × 8 9); Stieda body discoid; sub-Stieda body present; sporocyst residuum present, formed by a group of spheroid bodies between sporozoites. This study increases the number of viperid hosts of C. bigenetica and expands the geographical distribution to South America.

O coccídio Caryospora bigenetica foi descrito na serpente Crotalus horridus (Viperidae) nos Estados Unidos da América. Este estudo representa o primeiro registro da ocorrência de C. bigenetica em serpentes da América do Sul. Amostras de fezes foram obtidas, entre novembro de 2013 e maio de 2014, de 256 serpentes silvestres mantidas em um criatório científico nos Estados do Mato Grosso do Sul (MS; n = 214) e Rio de Janeiro (RJ; n = 42), Brasil. Caryospora bigenetica foi encontrada em 14 (5,6%) serpentes, todas pertencentes à família Viperidae. Dez Bothrops moojeni e duas Crotalus durissus de MS estavam infectadas. O coccídio também foi encontrado em uma C. durissus e uma Bothrops jararacussu do Estado do Rio de Janeiro. Os oocistos foram esféricos, com parede dupla, sendo a externa ligeiramente mamilonada, estriações aparentes transversalmente, 13.0 µm (12 14); grânulo polar junto à parede interna. Esporocisto oval ou piriforme, 10.0 × 8.0 µm (9 11 × 8 9); corpo de Stieda discóide; sub-Stieda presente; resíduo do esporocisto presente, formado por um grupo de corpos esféricos entre os esporozoítos. Este estudo aumenta o número de hospedeiros viperídeos de C. bigenetica e expande a distribuição geográfica para a América do Sul.

Determinação das concentrações plasmáticas de proteínas e metabólitos de cascavéis em cativeiro / Determination of proteins and metabolites concentrations in plasma of rattlesnakes in captivity

Para determinar as concentrações plasmáticas de proteínas e metabólitos de cascavéis em cativeiro, foram utilizadas 60 serpentes adultas, sendo 30 machos e 30 fêmeas. O sangue foi coletado através de punção do seio venoso paravertebral cervical e armazenado em tubos com heparina. As análises bioquímicas foram processadas colorimetricamente em Analisador Automático de Bioquímica Chemwell (Awareness Technology®, Inc). Foram calculadas as médias e desvios padrão dos seguintes constituintes: proteínas totais, albumina, globulinas, relação albumina/globulinas, ácido úrico, creatinina, ureia, colesterol, colesterol HDL e triglicérides. Os valores obtidos foram semelhantes aos descritos na literatura para repteis e serpentes, sendo as diferenças observadas provavelmente decorrentes da diferença entre espécies, clima, estação do ano e metodologia utilizada. Não houve diferenças significativas entre machos e fêmeas para os parâmetros estudados. Estes resultados podem ser úteis no estabelecimento de valores de referência para planos de conservação destes ofídios em cativeiro.(AU)

For determining plasma concentrations of proteins and metabolites of rattlesnakes in captivity, 60 adult snakes, 30 males and 30 females were used. Blood was collected by puncture of the cervical paravertebral venous sinus and stored in tubes with heparin. Biochemical analyzes were colorimetrically processed using an Automatic Biochemistry Analyzer Chemwell (Awareness Technology®, Inc). The mean and standard deviation were calculated for the following constituents: total protein, albumin, globulin, albumin/globulin ratio, uric acid, creatinine, urea, cholesterol, HDL cholesterol and triglycerides. The values were similar to those previously reported for reptiles and snakes, with the differences observed probably due to the difference between species, climate, season and the methodology used. There were no significant differences between males and females for the parameters studied. These results may be useful in establishing normal biochemical values for conservation plans for these snakes in captivity.(AU)

Perfil eco-epidemiológico de los accidentes por ofidios en Monagas, Venezuela (2002-2006) / Eco-epidemiological profile of snakebite accidents in Monagas state, Venezuela (2002-2006)

El accidente causado por animales venenosos representa un problema global, especialmente en las áreas subtropicales y tropicales del mundo. En Venezuela, es endémico para varias regiones incluyendo la zona nororiental. Con el objetivo de ampliar y actualizar el perfil eco-epidemiológico del ofidismo en el estado Monagas, se evaluó su comportamiento, en cada municipio, durante 5 años de registro (2002 - 2006). Se contabilizaron 339 accidentes ofídicos (con promedio de 68 anuales). El incidente ocurrió con mayor frecuencia en agricultores, adultos jóvenes y de género masculino, durante sus labores en el campo y en el periodo diurno. La incidencia siguió un patrón estacional bimodal con predominio en épocas de alta y baja pluviosidad. Los envenenamientos botrópicos (28,6%) fueron más frecuentes que los crotálicos (14,5%), con proporción 2:1. El 87,9% recibió sueroterapia específica. La incidencia promedio anual en la entidad se ubicó en 11,30 casos por 100.000 habitantes, siendo Punceres (46,29), Acosta (20,91) y Bolívar (19,52) los municipios con mayor impacto. El estado Monagas presentó un mapa de endemicidad distribuido en municipios de (1) muy alta endemicidad, (2) alta endemicidad, (3) mediana endemicidad, (4) baja endemicidad y (5) muy baja endemicidad. Durante el período analizado no se registró muerte por esta causa en el estado. Los hallazgos sugieren la importancia del accidente por serpientes en Monagas, con especial importancia en la mitad norte de la entidad.

The accidents caused by venomous animals are a global problem, especially in subtropical and tropical regions of the world. In Venezuela, they are endemic in several regions including the northeast. In order to expand and update the ecoepidemiological profile of snake bites in Monagas state, their behavior was assessed in each municipality for 5 years (2002 -2006). There were 339 ophidian accidents (on average 68 per year). The bites occurred more frequently in young adult male farmers, while working in the field and during daytime. The incidence followed a bimodal seasonal pattern with predominance in high and low rainfall periods. Bothropic envenoming (28.6%) were the most frequent followed by Crotalic ones (14.5%), with a 2:1 ratio. 87.9% received specific serum therapy. The annual average incidence in the state was 11.30 cases per 100,000 inhabitants. Punceres (46.29), Acosta (20.91) and Bolivar (19.52) were the municipalities with the highest impact. Monagas state showed an endemicity map with municipalities having (1) very high endemicity, (2) high endemicity, (3) medium endemicity, (4) low endemicity and (5) very low endemicity. In the studied period there were no deaths from this cause in the state. The findings suggest the importance of snake accidents in Monagas, especially in the northern half of the state.

Microhabitat use by species of the genera Bothrops and Crotalus (Viperidae) in semi-extensive captivity

Many factors influence microhabitat including climate and the occurrence of predators, prey and suitable shelters. The influence of predators in a semi-extensive breeding system is minimized due to frequent monitoring of the area. This situation enables the independent analysis of such other variables as refuges and temperature. Some specimens of the Viperidae family are kept in a semi-extensive breeding system at the Butantan Institute for display and study. These animals are widely distributed in the Atlantic Forest and Cerrado, two biomes with distinct climatic characteristics. We compared the daily activity pattern and microhabitat use of the species Bothrops jararaca and Crotalus durissus terrificus. Our main questions were whether rattlesnakes and lancehead snakes respond to habitat selection differently in similar climatic conditions and if they choose similar microhabitats. Species of the genus Bothrops were frequently found under shelters regardless of the time of day. On the other hand, snakes of the genus Crotalus were frequently found sheltered during the early morning, then migrated to sunnier areas and returned to shelters in late afternoon.

Characterization of venomous snakes of Thailand.

Background: Envenoming by snakebite is an important public health problem in rural tropics. Venomous snake families such as Elapidae and Viperidae frequently produce severe poisoning. Anti-venoms are not available for all venomous snakes of Thailand and there is need for more development in this field. Objective: We characterized the important venomous snakes’ distribution of Thailand. Method: Venomous snake species are described in details including their identification, range, and extraterritorial distribution. Result: Eighteen snake species of the family Elapidae are summarized in their characteristics and distribution. There are three species of Naja, one species of Ophiophagus, three species of Bungarus, four species of Calliophis, one species of Sinomicrurus, two species of Laticauda, and four species of subfamily Hydrophiinae. Fifteen snake species of the family Viperidae consisting of one species of subfamily Viperinae and fourteen species of subfamily Crotalinae are also discussed. Conclusion: All these snakes are venomous and their venom is potentially fatal since birth.

Amblyomma rotundatum (Koch, 1844) (Acari: Ixodidae) two-host life-cycle on Viperidae snakes / Ciclo dioxênico em Amblyomma rotundatum (Koch, 1844) (Acari: Ixodidae)parasitando serpentes da família Viperidae

Amblyomma rotundatum is an ixodid tick that infests ectothermic animals and reproduces exclusively by parthenogenesis. This tick has been frequently reported to infest reptiles and amphibians, under natural conditions and sometimes in captivity. It was described in Brazil and several other countries of South, Central and North America. Although many studies have reported aspects of its biology, none of them has used regularly either ophidian as hosts, or controlled temperature, humidity and luminosity for parasitic stages. The objective of this experiment was to study the life cycle of A. rotundatum feeding on Viperidae snakes under room controlled conditions at 27 ± 1 ºC temperature, 85 ± 10 percent relative humidity and 12:12 hours photoperiod for parasitic stages, and under B.O.D incubator conditions at 27 ± 1 ºC temperature, 85 ± 10 percent relative humidity and scotophase for non-parasitic stages. The total duration of the life cycle ranged from 56 to 163 days (mean of 105 days). Two-host life cycle was observed for most of the ixodid population studied.

Amblyomma rotundatum é um carrapato da família Ixodidae, parasito de animais pecilotérmicos, e que se reproduz exclusivamente por partenogênese. Este carrapato é frequentemente relatado infestando répteis e anfíbios em condições naturais e, às vezes, em animais de cativeiro. Ele já foi relatado no Brasil e em vários outros países das Américas do Sul, Central e do Norte. Embora muitos estudos sobre sua biologia tenham sido publicados, nunca foram utilizados ofídios como hospedeiros e, tão pouco, foram realizados ensaios com os estádios parasitários sob condições controladas de temperatura, umidade e iluminação. O objetivo deste experimento foi estudar o ciclo biológico de A. rotundatum se alimentando em serpentes da família Viperidae sob condições ambientais controladas a 27 ± 1 ºC de temperatura, 85 ± 10 por cento de umidade relativa do ar e 12:12 horas de fotoperíodo para estágios parasitários; assim como sob condições iguais a 27 ± 1 ºC de temperatura, 85 ± 10 por cento de umidade relativa do ar e escotofase em estufas de germinação para estádios não parasitários. A duração total do ciclo de vida variou de 56 a 163 dias (média de 105 dias). Observou-se ciclo dioxênico para a a maior parte da população dos ixodídeos em estudo.

The defensive strike of five species of lanceheads of the genus Bothrops (Viperidae)

We studied the defensive strike of one species of each of five recognized lineages within the genus Bothrops, namely, B. alternatus, B. jararaca, B. jararacussu, B. moojeni and B. pauloensis. The defensive strike of the studied species was in general similar to that of Crotalus viridis and C. atrox, but some important differences were observed. Bothrops alternatus and B. pauloensis struck preferentially from a tight body posture, whereas B. jararaca and B. moojeni from a loose body posture. Defensive strikes were either true or false (during the latter, the mouth remains closed or partially open). Almost all strikes were successful; only on a few occasions snakes missed their target (flawed strikes). Strike variables were very conservative among the five species, especially strike distance and height, and one possible explanation may be related to constraints imposed on strike variables as a way of increasing strike accuracy.

Estudamos o bote defensivo de uma espécie de cada uma de cinco reconhecidas linhagens do gênero Bothrops, a saber: B. alternatus, B. jararaca, B. jararacussu, B. moojeni e B. pauloensis. O bote defensivo das espécies estudadas foi, em geral, semelhante ao de Crotalus viridis e C. atrox, porém algumas diferenças foram observadas. Bothrops alternatus e B. pauloensis desferiram botes preferencialmente a partir de postura corpórea enrodilhada, ao passo que B. jararaca e B. moojeni desferiram a maioria dos botes a partir de postura corpórea frouxa. Os botes defensivos foram verdadeiros ou falsos (nestes, a boca da serpente permaneceu fechada ou parcialmente aberta). Quase todos os botes foram bem-sucedidos; apenas em alguns casos a serpente errou o alvo (botes falhos). As variáveis relativas aos botes foram bastante conservativas entre as cinco espécies, principalmente distância e altura do bote, e uma possível explicação pode estar relacionada a restrições impostas às variáveis relativas aos botes como forma de aumentar sua acurácia.

Caracterización Toxinológica del Veneno Total de la Serpiente de Cascabel Crotalus durissus cumanensis (VIPERIDAE), presente en la localidad de Porshoure, Guajira Venezolana / Toxinological Characterization of the Whole Venom of the Rattlesnake Crotalus durissus cumanensis (Viperidae) from Porshoure, Venezuelan Guajira

En el presente trabajo se caracterizó toxinológicamente el veneno total de la serpiente de cascabel Crotalus durissus cumanensis presente en la localidad de Porshoure, Guajira venezolana, determinando: dosis letal cincuenta (DL50), actividad hemorrágica, actividad edematizante, actividad hemolítica indirecta, actividad coagulante sobre plasma humano (in vitro) y actividad desfibrinante (in vivo), según las metodologías establecidas por la Organización Mundial de la Salud. Los resultados obtenidos mostraron que el veneno de esta subespecie es altamente tóxico, de naturaleza neurotóxica y coagulante. A nivel local produjo poca hemorragia y edema al compararlo con el veneno de otras serpientes de la misma subespecie, entre ellas Crotalus durissus cumanensis de la Villa del Rosario, estado Zulia; Crotalus durissus terrificus y Crotalus durissus durissus de Costa Rica y Brazil; Crotalus durissus terrificus de Colombia; Crotalus durissus ruruima, Crotalus durissus cascavella, Crotalus durissus collilineatus, entre otras. También resultó ser poco hemolítico y con características desfibrinantes (in vivo); efecto que hasta ahora no había sido reportado para serpientes pertenecientes al género Crotalus, lo que sin duda alguna constituye un valioso aporte para el diagnóstico y tratamiento del accidente crotálico en Venezuela.

The whole venom of the rattlesnake Crotalus durissus cumanensis (Viperidae) from Porshoure, Venezuelan Guajira, was characterized toxinologically in this research, determining: lethal dose 50 (LD50), bleeding, edematic and hemolytic activity, coagulant activity on human plasma (in vitro) and fibrinolytic activity, according to the methodologies outlined by the World Health Organization. The results demonstrated that the venom of the studied species is highly toxic, neurotoxic and coagulant. Local effects such as the hemorrhage and edema were not highly marked in comparison with other snakes’ venoms of the same subspecies as Crotalus durissus cumanensis from La Villa del Rosario, Zulia state; Crotalus durissus terrificus from Costa Rica and Crotalus durissus durissus from Brazil; Crotalus durissus terrificus from Colombia and Crotalus durissus ruruima, Crotalus durissus cascavella and Crotalus durissus collilineatus. The venom was slightly hemolytic and with fibrinolytic characteristic (in vivo). This effect has not yet been described within the genus Crotalus, what is a significant addition to the information of diagnosis and treatment of the rattlesnake bites in Venezuela.