Taiwan cobra envenoming: serum venom concentration before and after specific treatment and relationship with debridement of necrotic wound tissue

Background: Bivalent freeze-dried neurotoxic (FN) antivenom has been the primary treatment since the 1980s for Taiwan cobra (Naja atra) envenomation in Taiwan. However, envenomation-related wound necrosis is a significant problem after cobra snakebites. In the present study, we analyzed the changes in serum venom concentration before and after antivenom administration to discover their clinical implications and the surgical treatment options for wound necrosis. Methods: The patients were divided into limb swelling and wound necrosis groups. The clinical outcome was that swelling started to subside 12 hours after antivenom treatment in the first group. Serum venom concentrations before and after using antivenoms were measured to assess the antivenom's ability to neutralize the circulating cobra venom. The venom levels in wound wet dressing gauzes, blister fluids, and debrided tissues were also investigated to determine their clinical significance. We also observed the evolutional changes of wound necrosis and chose a better wound debridement timing. Results: We prospectively enrolled 15 Taiwan cobra snakebite patients. Males accounted for most of this study population (n = 11, 73%). The wound necrosis group received more antivenom doses than the limb swelling group (4; IQR:2-6 vs 1; IQR:1-2, p = 0.05), and less records of serum venom concentrations changed before/after antivenom use (p = 0.0079). The necrotic wound site may release venom into circulation and cause more severe envenomation symptoms. Antivenom can efficiently diminish limb swelling in cobra bite patients. However, antivenom cannot reduce wound necrosis. Patients with early debridement of wound necrosis had a better limb outcome, while late or without debridement may have long-term hospital stay and distal limb morbidity. Conclusions: Antivenom can efficiently eliminate the circulating cobra venom in limb swelling patients without wound necrosis. Early debridement of the bite site wound and wet dressing management are suggestions for preventing extended tissue necrosis and hospital stay.(AU)

S- and P-type cobra venom cardiotoxins differ in their action on isolated rat heart

Background: The cardiovascular system is one of the first systems to be affected by snake toxins; but not many toxins exert a direct effect on the heart. Cobra venom cardiotoxins are among those few toxins that attack the heart. Although the two cardiotoxin types (S and P) differ in their central-loop structure, it is not known whether they differ in their effect on the mammalian heart. We compared the effects of S- and P-type cardiotoxins, CTÐ¥-1 and CTÐ¥-2, respectively, from the cobra Naja oxiana, on the isolated rat heart. Methods: An isolated rat heart perfused according to the Langendorff technique was used in this study to investigate the activity of cardiotoxins CTX-1 and CTX-2. The following parameters were registered: the left ventricular developed pressure, calculated as the difference between systolic and diastolic pressure in the left ventricle, the end-diastolic pressure, the heart rate, time to maximal end-diastolic pressure (heart contracture), and time to depression of the heart contraction. Results: Both cardiotoxins at the concentration of 5 µg/mL initially produce a slight increase in systolic intraventricular pressure, followed by its rapid decrease with a simultaneous increase in diastolic intraventricular pressure until reaching contracture. CTX-2 blocks cardiac contractions faster than CTX-1; in its presence the maximum diastolic pressure is reached faster and the magnitude of the developed contracture is higher. Conclusion: The P-type cardiotoxin CTX-2 more strongly impairs rat heart functional activity than the S-type cardiotoxin CTX-1, as expressed in its faster blockage of cardiac contractions as well as in more rapid development and greater magnitude of contracture in its presence.(AU)

Guidelines for the diagnosis and differential diagnosis of snakebite envenoming of ruminants in Brazil / Diretrizes para o diagnóstico e diagnóstico diferencial de envenenamento por picadas de cobra em ruminantes no Brasil

In Brazil, snakebites are often cited as a cause of mortality in ruminants, but there are discrepancies in the literature regarding its actual prevalence, either by lack of diagnosis or by mistakes in the differential diagnosis. Among the factors that hinder the diagnosis are included the inconsistencies to distinguish between accidents caused by Bothrops and Crotalus, responsible for over 90% of the cases. For the diagnosis of accidents involving Lachesis muta, both the neurotropic and the proteolytic/hemolytic effects must be considered, similar to what is described in Crotalus scutulatus. This article describes the main clinical, pathological, and laboratory findings observed in envenoming by the aforementioned snakes and suggests procedures for establishing the diagnosis and differential diagnosis starting from a logical sequence, based on epidemiological evidence, clinical, laboratory, and pathological findings.

No Brasil, acidentes ofídicos são frequentemente citados como causa de mortalidade em ruminantes, mas existem discrepâncias em relação a sua atual prevalência, seja por falta de diagnóstico ou por erros no diagnóstico diferencial. Entre os fatores que dificultam o diagnóstico estão as inconsistências para distinguir entre os acidentes causados por Bothrops e Crotalus, responsáveis por mais de 90% dos casos. Para o diagnóstico de envenenamentos por Lachesis muta, devem ser considerados os efeitos neurotrópico e proteolítico/hemolíticos concomitantes, a exemplo do que ocorre com algumas cascavéis norte-americanas (Crotalus scutulatus, entre elas). Este artigo descreve os principais achados clinicopatológicos e laboratoriais observados em casos de envenenamento pelas serpentes citadas e sugere um roteiro simplificado para o estabelecimento do diagnóstico e diagnóstico diferencial, a partir de uma sequência lógica, baseada em evidências epidemiológicas e achados clínicos, laboratoriais e patológicos.

So far from home: first record of Chondromorpha xanthotricha (Diplopoda: Polydesmida: Paradoxosomatidae) in the Brazilian Amazon / Tão longe de casa: primeiro registro de Chondromorpha xanthotricha (Diplopoda: Polydesmida: Paradoxosomatidae) para a Amazônia brasileira

The millipede Chondromorpha xanthotricha, supposedly native to Sri Lanka and southern India, is considered a pantropical species occurring in Southeast Asia, North America, Mesoamerica, the Caribbean, Central America and northern South America. Here we report the first record of this species for Brazil, in the central Amazon region of the country, with taxonomic notes and images of male and female specimens.(AU)

O diplópode Chondromorpha xanthotricha, supostamente nativo do Sri Lanka e sul da Índia, é considerado uma espécie pantropical com ocorrência no Sudeste Asiático, América do Norte, Mesoamérica, Caribe, América Central e norte da América do Sul. Aqui nós reportamos o primeiro registro dessa espécie para o Brasil, na região central da Amazônia brasileira, com notas taxonômicas e ilustrações de espécimes macho e fêmea.(AU)

Proteomic characterization of Naja mandalayensis venom

Background Naja mandalayensis is a spitting cobra from Myanmar. To the best of our knowledge, no studies on this venom composition have been conducted so far. On the other hand, few envenomation descriptions state that it elicits mainly local inflammation in the victims' eyes, the preferred target of this spiting cobra. Symptoms would typically include burning and painful sensation, conjunctivitis, edema and temporary loss of vision. Methods We have performed a liquid-chromatography (C18-RP-HPLC) mass spectrometry (ESI-IT-TOF/MS) based approach in order to biochemically characterize N. mandalayensis venom. Results A wide variety of three-finger toxins (cardiotoxins) and metallopeptidases were detected. Less abundant, but still representative, were cysteine-rich secretory proteins, L-amino-acid oxidases, phospholipases A2, venom 5'-nucleotidase and a serine peptidase inhibitor. Other proteins were present, but were detected in a relatively small concentration. Conclusion The present study set the basis for a better comprehension of the envenomation from a molecular perspective and, by increasing the interest and information available for this species, allows future venom comparisons among cobras and their diverse venom proteins.(AU)

Proteomic characterization of Naja mandalayensis venom

Naja mandalayensis is a spitting cobra from Myanmar. To the best of our knowledge, no studies on this venom composition have been conducted so far. On the other hand, few envenomation descriptions state that it elicits mainly local inflammation in the victims' eyes, the preferred target of this spiting cobra. Symptoms would typically include burning and painful sensation, conjunctivitis, edema and temporary loss of vision. Methods We have performed a liquid-chromatography (C18-RP-HPLC) mass spectrometry (ESI-IT-TOF/MS) based approach in order to biochemically characterize N. mandalayensis venom. Results A wide variety of three-finger toxins (cardiotoxins) and metallopeptidases were detected. Less abundant, but still representative, were cysteine-rich secretory proteins, L-amino-acid oxidases, phospholipases A2, venom 5'-nucleotidase and a serine peptidase inhibitor. Other proteins were present, but were detected in a relatively small concentration. Conclusion The present study set the basis for a better comprehension of the envenomation from a molecular perspective and, by increasing the interest and information available for this species, allows future venom comparisons among cobras and their diverse venom proteins.(AU)

Antimicrobial effect of bothropstoxin-i in broilers / Atividade antimicrobiana da bothropstoxin-i em frangos de corte

Bacterial resistance is a sanitary issue explained by indiscriminate use of nonprescription drugs, and antimicrobial use in food production for growth promotion. Bothropstoxin-I (BthTx-I) is a phospholipase A2 (PLA2) from Bothrops jararacussu venom, which has a known antimicrobial effect. The goal of this study was the unprecedented evaluation of in vivo antimicrobial activity of BthTx-I in broilers. Microbiological, biochemical, and histological parameters were determined using 84 21-day old broilers that were kept in cages with four birds each at a density of 625 cm2/broiler. The experiment was randomized by three treatments with seven repetitions of four broilers each that lasted seven days. The treatments were: 1) bacitracin zinc diet; 2) PLA2-BthTx-I; 3) without additives. The data obtained from the studied variables was subjected to analysis of variance and an F-test at the 5% significance level. Averages of each variable in each treatment were compared by Tukey’s test. Broiler bacterial cloacal counts showed that BthTx-I decreased the microbial population without reducing body weight, intestinal morphology, or liver or kidney histopathological damage. The toxin showed in vivo activity, being an alternative for better performance in the production of broiler chickens, because it acted by decreasing the microbial load of potentially pathogenic bacteria in the intestinal(AU)

A resistência bacteriana é uma questão sanitária, explicada pelo uso indiscriminado de medicamentos sem receita médica e pelo uso de antimicrobianos na produção de alimentos para promover o crescimento. Bothropstoxin-I (BthTx-I) é uma fosfolipase A2 (PLA2) obtida do veneno da Bothrops jararacussu. A PLA2 do veneno de cobra tem efeito antimicrobiano conhecido. Objetivou-se com este estudo avaliar sem precedentes a atividade antimicrobiana in vivo de BthTx-I em frangos de corte. Os parâmetros microbiológicos, bioquímicos e histológicos foram realizados em 84 frangos de corte com 21 dias de idade mantidos em gaiolas com quatro animais cada e densidade de 625 cm2/frango. O experimento foi dividido em três tratamentos com sete repetições de quatro frangos cada um, com duração de sete dias. Os tratamentos foram: 1) dieta com bacitracina de zinco; 2) PLA2-BthTx-I; 3) sem aditivos. Os dados obtidos das variáveis estudadas foram submetidos à análise de variância e teste F ao nível de significância de 5%. As médias dos tratamentos em cada variável foram comparadas pelo teste de Tukey. A contagem cloacal bacteriana de frangos de corte mostrou que o BthTx-I diminuiu a população microbiana sem comprometer o peso corporal, a morfologia intestinal ou causar danos histopatológico no fígado e rins. Concluiu-se que a toxina apresentou atividade in vivo, sendo uma alternativa para um melhor desempenho na produção de frangos de corte, pois agiu diminuindo a carga microbiana de bactérias potencialmente patogênicas na microbiota intestinal das aves e não causou danos musculares, hepáticos ou renais na dosagem avaliada.(AU)

Antimicrobial effect of bothropstoxin-i in broilers / Atividade antimicrobiana da bothropstoxin-i em frangos de corte

Bacterial resistance is a sanitary issue explained by indiscriminate use of nonprescription drugs, and antimicrobial use in food production for growth promotion. Bothropstoxin-I (BthTx-I) is a phospholipase A2 (PLA2) from Bothrops jararacussu venom, which has a known antimicrobial effect. The goal of this study was the unprecedented evaluation of in vivo antimicrobial activity of BthTx-I in broilers. Microbiological, biochemical, and histological parameters were determined using 84 21-day old broilers that were kept in cages with four birds each at a density of 625 cm2/broiler. The experiment was randomized by three treatments with seven repetitions of four broilers each that lasted seven days. The treatments were: 1) bacitracin zinc diet; 2) PLA2-BthTx-I; 3) without additives. The data obtained from the studied variables was subjected to analysis of variance and an F-test at the 5% significance level. Averages of each variable in each treatment were compared by Tukey’s test. Broiler bacterial cloacal counts showed that BthTx-I decreased the microbial population without reducing body weight, intestinal morphology, or liver or kidney histopathological damage. The toxin showed in vivo activity, being an alternative for better performance in the production of broiler chickens, because it acted by decreasing the microbial load of potentially pathogenic bacteria in the intestinal

A resistência bacteriana é uma questão sanitária, explicada pelo uso indiscriminado de medicamentos sem receita médica e pelo uso de antimicrobianos na produção de alimentos para promover o crescimento. Bothropstoxin-I (BthTx-I) é uma fosfolipase A2 (PLA2) obtida do veneno da Bothrops jararacussu. A PLA2 do veneno de cobra tem efeito antimicrobiano conhecido. Objetivou-se com este estudo avaliar sem precedentes a atividade antimicrobiana in vivo de BthTx-I em frangos de corte. Os parâmetros microbiológicos, bioquímicos e histológicos foram realizados em 84 frangos de corte com 21 dias de idade mantidos em gaiolas com quatro animais cada e densidade de 625 cm2/frango. O experimento foi dividido em três tratamentos com sete repetições de quatro frangos cada um, com duração de sete dias. Os tratamentos foram: 1) dieta com bacitracina de zinco; 2) PLA2-BthTx-I; 3) sem aditivos. Os dados obtidos das variáveis estudadas foram submetidos à análise de variância e teste F ao nível de significância de 5%. As médias dos tratamentos em cada variável foram comparadas pelo teste de Tukey. A contagem cloacal bacteriana de frangos de corte mostrou que o BthTx-I diminuiu a população microbiana sem comprometer o peso corporal, a morfologia intestinal ou causar danos histopatológico no fígado e rins. Concluiu-se que a toxina apresentou atividade in vivo, sendo uma alternativa para um melhor desempenho na produção de frangos de corte, pois agiu diminuindo a carga microbiana de bactérias potencialmente patogênicas na microbiota intestinal das aves e não causou danos musculares, hepáticos ou renais na dosagem avaliada.

Clinical and post mortem examination of white worm lizards (Amphisbaena alba) in the State of Paraíba, Northeastern Brazil: morphological, pathological and radiographic findings of a secretive species

ABSTRACT: Amphisbenians are limbless reptiles that belong to the order Squamata. Due to their fossorial and secrevie habits, little is known about their morphology, ecology and pathological conditions that may affect them. In this manuscript, we present a brief guide for identification of normal structures as well as traumatic injuries on radiography and necropsy of Amphisbaena alba. From April to September 2019, three cases of A. alba with suspected trauma were referred to the Veterinary Hospital of the Federal University of Paraíba (UFPB). In the clinical evaluation, traumatic injuries were observed, and support therapy was instituted, but they did not resist and died shortly after. Bone fractures and organ ruptures, in addition to specific structures of this species were identified on radiography. A systematic necropsy was performed of all amphisbaenians in order to evaluate external and internal structures, not only to identify lesions but also to investigate the morphological aspects of amphisbenids. Macroscopically, multiple organ fractures and ruptures observed in radiographs were confirmed, in addition to the presence of the cestodes Semenoviella amphisbaenae in the large intestine. Histologically, it was possible to identify normal characteristics and microscopic lesions in the tissues. This is the first study to incorporate morphological, clinical, and pathological aspects of A. alba. This manuscript brings essential information for wildlife veterinarians and pathologists who may have to treat or perform a necropsy on these unique reptiles.

RESUMO: Anfisbenas são répteis desprovidos de membros que pertencem a ordem Squamata. Devido a seus hábitos fossoriais e reclusos, pouco se conhece sobre sua morfologia, ecologia e condições patológicas que possam afetá-las. Neste artigo, um guia sucinto para a necropsia, identificação de tecidos e órgãos e lesões encontradas em Amphisbaena alba, é apresentado. Durante março e dezembro de 2019, três casos de A. alba com suspeita de trauma foram encaminhados ao Hospital Veterinário da Universidade Federal da Paraíba. Na avaliação clínica foram observadas lesões traumáticas e instituído um suporte terapêutico, porém não resistiram e morreram em seguida. Foram realizadas projeções radiográficas onde identificaram fraturas e ruptura de órgãos, além de identificar estruturas específicas dessa espécie. Uma avaliação completa de estruturas externas e internas foi conduzida para investigar aspectos morfológicos dos anfisbenídeos. Macroscopicamente foram confirmadas múltiplas fraturas e rupturas de órgãos observadas anteriormente nas radiografias, além disso evidenciou no intestino grosso presença de parasitos anoplocéfalos denominados Semenoviella amphisbaenae. Na histologia foi possível identificar características normais e lesões microscópicas nos tecidos. Esse é o primeiro estudo a incorporar aspectos morfológicos, clínicos e patológicos de A. alba. Esse manuscrito traz informações essenciais para clínicos e patologistas de animais selvagens que podem se deparar com casos clínicos ou de necropsia de Amphisbaena alba.

Clinical and post mortem examination of white worm lizards (Amphisbaena alba) in the State of Paraíba, Northeastern Brazil: morphological, pathological and radiographic findings of a secretive species / Exame clínico e post mortem de anfisbenas (Amphisbaena alba) no Estado da Paraíba, Nordeste do Brasil: achados morfológicos, patológicos e radiográficos de uma espécie reservada

Amphisbenians are limbless reptiles that belong to the order Squamata. Due to their fossorial and secrevie habits, little is known about their morphology, ecology and pathological conditions that may affect them. In this manuscript, we present a brief guide for identification of normal structures as well as traumatic injuries on radiography and necropsy of Amphisbaena alba. From April to September 2019, three cases of A. alba with suspected trauma were referred to the Veterinary Hospital of the Federal University of Paraíba (UFPB). In the clinical evaluation, traumatic injuries were observed, and support therapy was instituted, but they did not resist and died shortly after. Bone fractures and organ ruptures, in addition to specific structures of this species were identified on radiography. A systematic necropsy was performed of all amphisbaenians in order to evaluate external and internal structures, not only to identify lesions but also to investigate the morphological aspects of amphisbenids. Macroscopically, multiple organ fractures and ruptures observed in radiographs were confirmed, in addition to the presence of the cestodes Semenoviella amphisbaenae in the large intestine. Histologically, it was possible to identify normal characteristics and microscopic lesions in the tissues. This is the first study to incorporate morphological, clinical, and pathological aspects of A. alba. This manuscript brings essential information for wildlife veterinarians and pathologists who may have to treat or perform a necropsy on these unique reptiles.(AU)

Anfisbenas são répteis desprovidos de membros que pertencem a ordem Squamata. Devido a seus hábitos fossoriais e reclusos, pouco se conhece sobre sua morfologia, ecologia e condições patológicas que possam afetá-las. Neste artigo, um guia sucinto para a necropsia, identificação de tecidos e órgãos e lesões encontradas em Amphisbaena alba, é apresentado. Durante março e dezembro de 2019, três casos de A. alba com suspeita de trauma foram encaminhados ao Hospital Veterinário da Universidade Federal da Paraíba. Na avaliação clínica foram observadas lesões traumáticas e instituído um suporte terapêutico, porém não resistiram e morreram em seguida. Foram realizadas projeções radiográficas onde identificaram fraturas e ruptura de órgãos, além de identificar estruturas específicas dessa espécie. Uma avaliação completa de estruturas externas e internas foi conduzida para investigar aspectos morfológicos dos anfisbenídeos. Macroscopicamente foram confirmadas múltiplas fraturas e rupturas de órgãos observadas anteriormente nas radiografias, além disso evidenciou no intestino grosso presença de parasitos anoplocéfalos denominados Semenoviella amphisbaenae. Na histologia foi possível identificar características normais e lesões microscópicas nos tecidos. Esse é o primeiro estudo a incorporar aspectos morfológicos, clínicos e patológicos de A. alba. Esse manuscrito traz informações essenciais para clínicos e patologistas de animais selvagens que podem se deparar com casos clínicos ou de necropsia de Amphisbaena alba.(AU)

Clinical and post mortem examination of white worm lizards (Amphisbaena alba) in the State of Paraíba, Northeastern Brazil: morphological, pathological and radiographic findings of a secretive species / Exame clínico e post mortem de anfisbenas (Amphisbaena alba) no Estado da Paraíba, Nordeste do Brasil: achados morfológicos, patológicos e radiográficos de uma espécie reservada

Amphisbenians are limbless reptiles that belong to the order Squamata. Due to their fossorial and secrevie habits, little is known about their morphology, ecology and pathological conditions that may affect them. In this manuscript, we present a brief guide for identification of normal structures as well as traumatic injuries on radiography and necropsy of Amphisbaena alba. From April to September 2019, three cases of A. alba with suspected trauma were referred to the Veterinary Hospital of the Federal University of Paraíba (UFPB). In the clinical evaluation, traumatic injuries were observed, and support therapy was instituted, but they did not resist and died shortly after. Bone fractures and organ ruptures, in addition to specific structures of this species were identified on radiography. A systematic necropsy was performed of all amphisbaenians in order to evaluate external and internal structures, not only to identify lesions but also to investigate the morphological aspects of amphisbenids. Macroscopically, multiple organ fractures and ruptures observed in radiographs were confirmed, in addition to the presence of the cestodes Semenoviella amphisbaenae in the large intestine. Histologically, it was possible to identify normal characteristics and microscopic lesions in the tissues. This is the first study to incorporate morphological, clinical, and pathological aspects of A. alba. This manuscript brings essential information for wildlife veterinarians and pathologists who may have to treat or perform a necropsy on these unique reptiles.(AU)

Anfisbenas são répteis desprovidos de membros que pertencem a ordem Squamata. Devido a seus hábitos fossoriais e reclusos, pouco se conhece sobre sua morfologia, ecologia e condições patológicas que possam afetá-las. Neste artigo, um guia sucinto para a necropsia, identificação de tecidos e órgãos e lesões encontradas em Amphisbaena alba, é apresentado. Durante março e dezembro de 2019, três casos de A. alba com suspeita de trauma foram encaminhados ao Hospital Veterinário da Universidade Federal da Paraíba. Na avaliação clínica foram observadas lesões traumáticas e instituído um suporte terapêutico, porém não resistiram e morreram em seguida. Foram realizadas projeções radiográficas onde identificaram fraturas e ruptura de órgãos, além de identificar estruturas específicas dessa espécie. Uma avaliação completa de estruturas externas e internas foi conduzida para investigar aspectos morfológicos dos anfisbenídeos. Macroscopicamente foram confirmadas múltiplas fraturas e rupturas de órgãos observadas anteriormente nas radiografias, além disso evidenciou no intestino grosso presença de parasitos anoplocéfalos denominados Semenoviella amphisbaenae. Na histologia foi possível identificar características normais e lesões microscópicas nos tecidos. Esse é o primeiro estudo a incorporar aspectos morfológicos, clínicos e patológicos de A. alba. Esse manuscrito traz informações essenciais para clínicos e patologistas de animais selvagens que podem se deparar com casos clínicos ou de necropsia de Amphisbaena alba.(AU)

Suppression of cardiomyocyte functions by ß-CTX isolated from the Thai king cobra (Ophiophagus hannah) venom via an alternative method

Beta-cardiotoxin (ß-CTX), the three-finger toxin isolated from king cobra (Ophiophagus hannah) venom, possesses ß-blocker activity as indicated by its negative chronotropy and its binding property to both ß-1 and ß-2 adrenergic receptors and has been proposed as a novel ß-blocker candidate. Previously, ß-CTX was isolated and purified by FPLC. Here, we present an alternative method to purify this toxin. In addition, we tested its cytotoxicity against different mammalian muscle cell types and determined the impact on cardiac function in isolated cardiac myocyte so as to provide insights into the pharmacological action of this protein. Methods: ß-CTX was isolated from the crude venom of the Thai king cobra using reverse-phased and cation exchange HPLC. In vitro cellular viability MTT assays were performed on mouse myoblast (C2C12), rat smooth muscle (A7r5), and rat cardiac myoblast (H9c2) cells. Cell shortening and calcium transient dynamics were recorded on isolated rat cardiac myocytes over a range of ß-CTX concentration. Results: Purified ß-CTX was recovered from crude venom (0.53% w/w). MTT assays revealed 50% cytotoxicity on A7r5 cells at 9.41 ± 1.14 µM (n = 3), but no cytotoxicity on C2C12 and H9c2 cells up to 114.09 µM. ß-CTX suppressed the extend of rat cardiac cell shortening in a dose-dependent manner; the half-maximal inhibition concentration was 95.97 ± 50.10 nM (n = 3). In addition, the rates of cell shortening and re-lengthening were decreased in ß-CTX treated myocytes concomitant with a prolongation of the intracellular calcium transient decay, indicating depression of cardiac contractility secondary to altered cardiac calcium homeostasis. Conclusion: We present an alternative purification method for ß-CTX from king cobra venom. We reveal cytotoxicity towards smooth muscle and depression of cardiac contractility by this protein. These data are useful to aid future development of pharmacological agents derived from ß-CTX.(AU)

Suppression of cardiomyocyte functions by ß-CTX isolated from the Thai king cobra (Ophiophagus hannah) venom via an alternative method

Beta-cardiotoxin (ß-CTX), the three-finger toxin isolated from king cobra (Ophiophagus hannah) venom, possesses ß-blocker activity as indicated by its negative chronotropy and its binding property to both ß-1 and ß-2 adrenergic receptors and has been proposed as a novel ß-blocker candidate. Previously, ß-CTX was isolated and purified by FPLC. Here, we present an alternative method to purify this toxin. In addition, we tested its cytotoxicity against different mammalian muscle cell types and determined the impact on cardiac function in isolated cardiac myocyte so as to provide insights into the pharmacological action of this protein. Methods: ß-CTX was isolated from the crude venom of the Thai king cobra using reverse-phased and cation exchange HPLC. In vitro cellular viability MTT assays were performed on mouse myoblast (C2C12), rat smooth muscle (A7r5), and rat cardiac myoblast (H9c2) cells. Cell shortening and calcium transient dynamics were recorded on isolated rat cardiac myocytes over a range of ß-CTX concentration. Results: Purified ß-CTX was recovered from crude venom (0.53% w/w). MTT assays revealed 50% cytotoxicity on A7r5 cells at 9.41 ± 1.14 µM (n = 3), but no cytotoxicity on C2C12 and H9c2 cells up to 114.09 µM. ß-CTX suppressed the extend of rat cardiac cell shortening in a dose-dependent manner; the half-maximal inhibition concentration was 95.97 ± 50.10 nM (n = 3). In addition, the rates of cell shortening and re-lengthening were decreased in ß-CTX treated myocytes concomitant with a prolongation of the intracellular calcium transient decay, indicating depression of cardiac contractility secondary to altered cardiac calcium homeostasis. Conclusion: We present an alternative purification method for ß-CTX from king cobra venom. We reveal cytotoxicity towards smooth muscle and depression of cardiac contractility by this protein. These data are useful to aid future development of pharmacological agents derived from ß-CTX.(AU)

Hepatozoon pyramidumi sp. n. (Apicomplexa: Adeleorina) from the blood of Echis pyramidum: morphology and SSU rDNA sequence / Hepatozoon pyramidumi sp. n. (Apicomplexa: Adeleorina) do sangue de Echis pyramidum: morfologia e sequência de SSU rDNA

Hepatozoon pyramidumi sp. n. is described from the blood of the Egyptian saw-scaled viper, Echis pyramidum, captured from Saudi Arabia. Five out of ten viper specimens examined (50%) were found infected with Hepatozoon pyramidumi sp. n. with parasitaemia level ranged from 20-30%. The infection was restricted only to the erythrocytes. Two morphologically different forms of intraerythrocytic stages were observed; small and mature gamonts. The small ganomt with average size of 10.7 × 3.5 μm. Mature gamont was sausage-shaped with recurved poles measuring 16.3 × 4.2 μm in average size. Infected erythrocytes were hypertrophied; their nuclei were deformed and sometimes displaced from their central position in the normal uninfected cell. Merogonic stages were observed in the lung endothelial cell and the liver parenchyma cells. Mature meront was 17.8 × 13.6 µm and contained banana-shaped merozoites with average size of ~15 × 2 µm. Phylogenetic analysis based on the SSU rDNA sequence clustered Hepatozoon pyramidumi sp. n with previously sequenced Hepatozoon spp., most of them infected reptilian hosts without geographic consideration. The morphological and molecular comparison with closely related species proved the taxonomic uniqueness and novelty of the present form.(AU)

Hepatozoon pyramidumi sp. n. é descrito a partir do sangue da víbora em escamas e quilhas serrilhadas, Echis pyramidum, capturada na Arábia Saudita. Cinco de dez espécimes de víbora examinadas (50%) foram encontradas infectadas com Hepatozoon pyramidumi sp. n. com nível de parasitemia de 20% a 30%. A infecção foi restrita apenas aos eritrócitos. Foram observadas duas formas morfologicamente diferentes de estágios intra-eritrocíticos: gamontes de tamanho pequeno e madura. As formas menores de gamontes apresentaram média de 10,7 × 3,5 μm. Os gamontes maduros apresentaram forma de salsicha, com pequenos polos recurvados, medindo 16,3 × 4,2 μm, em média. Os eritrócitos infectados estavam aumentados de tamanho; seus núcleos encontravam-se deformados e, algumas vezes, deslocados de sua posição central, quando comparados às células normais não-infectadas. Foram observados estágios merogônicos em células endoteliais pulmonares e nas células do parênquima hepático. Os merontes maduros apresentavam 17,8 × 13,6 µm e continham merozoítos em forma de banana com tamanho médio de ~ 15 × 2 µm. A análise filogenética baseada nas sequências SSU rDNA agrupou Hepatozoon pyramidumi sp. n com Hepatozoon spp. detectados em répteis de várias regiões geográficas. Por meio de análises morfológicas e moleculares com espécies intimamente relacionadas, demonstrou-se a singularidade dessa nova espécie de Hepatozoon.(AU)

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?

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 120g of B. alternatus venom, while G3 received 200L 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.(AU)

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 120g de veneno de B. alternatus, enquanto o G3 recebeu apenas 200L 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.(AU)

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?

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 120g of B. alternatus venom, while G3 received 200L 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.

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 120g de veneno de B. alternatus, enquanto o G3 recebeu apenas 200L 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.

Enucleação em serpente urutu (Bothrops alternatus) no tratamento de endoftalmite decorrente de traumatismo de escama corneal / Urutu (Bothrops alternatus) snake enucleation in the treatment of endophthalmitis resulting from spectacle trauma

A urutu, Bothrops alternatus, é uma espécie de serpente peçonhenta da família Viperidae e subfamília Crotalinae. Como em outras espécies, ela pode ser acometida por acidentes ofídicos em algumas situações. Relata-se o caso de uma urutu que teve a escama corneal traumatizada em decorrência do ataque de outra cobra do mesmo gênero há 30 dias. Ao exame oftálmico, notou-se a presença de crosta recobrindo exsudato caseoso localizado no espaço subespetacular. Durante a exploração cirúrgica, foi removida a escama corneal e o exsudato caseoso, notando-se o quadro de endoftalmite. Procedeu-se à enucleação do bulbo acometido e o animal se recuperou bem. Diante de alguns casos, especificamente os que envolvem animais peçonhentos, o tratamento clínico com sua manipulação várias vezes ao dia torna-se inviável, sendo recomendado um tratamento cirúrgico definitivo.

The urutu (Bothrops alternatus) belongs to the Viperidae family and to the subfamily of venomous vipers, the Crotalinae. As with all other species the urutu is also susceptible to ophidian accidents. This is a case report of an urutu snake brought to the veterinary hospital with an injured spectacle. According to the clinical history, this lesion was caused by the attack of another snake of the same genus about 30 days earlier. Ophthalmic examination revealed a crust overlaying cheesy exudates just in the subspectacle space. After removal of these from the eye, an endophthalmitis was discovered. Enucleation of the affected ocular globe was performed allowing full recovery of the animal. Poisonous animals are difficult to handle, and I are important limitation to managements that require prolonged and multiple treatments throughout the day. Hence, in such cases, the preference for radical surgical procedures might be justifiable.

Enucleação em serpente urutu (Bothrops alternatus) no tratamento de endoftalmite decorrente de traumatismo de escama corneal / Urutu (Bothrops alternatus) snake enucleation in the treatment of endophthalmitis resulting from spectacle trauma

A urutu, Bothrops alternatus, é uma espécie de serpente peçonhenta da família Viperidae e subfamília Crotalinae. Como em outras espécies, ela pode ser acometida por acidentes ofídicos em algumas situações. Relata-se o caso de uma urutu que teve a escama corneal traumatizada em decorrência do ataque de outra cobra do mesmo gênero há 30 dias. Ao exame oftálmico, notou-se a presença de crosta recobrindo exsudato caseoso localizado no espaço subespetacular. Durante a exploração cirúrgica, foi removida a escama corneal e o exsudato caseoso, notando-se o quadro de endoftalmite. Procedeu-se à enucleação do bulbo acometido e o animal se recuperou bem. Diante de alguns casos, especificamente os que envolvem animais peçonhentos, o tratamento clínico com sua manipulação várias vezes ao dia torna-se inviável, sendo recomendado um tratamento cirúrgico definitivo.(AU)

The urutu (Bothrops alternatus) belongs to the Viperidae family and to the subfamily of venomous vipers, the Crotalinae. As with all other species the urutu is also susceptible to ophidian accidents. This is a case report of an urutu snake brought to the veterinary hospital with an injured spectacle. According to the clinical history, this lesion was caused by the attack of another snake of the same genus about 30 days earlier. Ophthalmic examination revealed a crust overlaying cheesy exudates just in the subspectacle space. After removal of these from the eye, an endophthalmitis was discovered. Enucleation of the affected ocular globe was performed allowing full recovery of the animal. Poisonous animals are difficult to handle, and I are important limitation to managements that require prolonged and multiple treatments throughout the day. Hence, in such cases, the preference for radical surgical procedures might be justifiable.(AU)

Acidente ofídico em um cão - relato de caso / Ophidian accident in a dog - case report

Foi atendido no Hospital Veterinário da Universidade Federal do Rio Grande do Sul, um canino, da raça Bulldog, de oito anos, pesando 32 kg, o qual habitava em um sítio. Foi relatado que o animal havia sofrido um acidente ofídico há duas horas e recebeu soro antiofídico em doses não conhecidas, por via subcutânea, cerca de uma hora após o acidente. A cobra foi descrita como da espécie Bothrops alternatus. O exame clínico mostrou edema acentuado na região submandibular direita, hematoma e lesões com sangramento no ponto de inoculação no pescoço. O animal foi hospitalizado e foram realizados hemograma e exames bioquímicos (ALT, creatinina, ureia), os quais não tiveram alterações, além de testes de coagulação (TP, TTPa), os quais se mostraram incoaguláveis (acima de 30s).

Acidente ofídico em um cão - relato de caso / Ophidian accident in a dog - case report

Foi atendido no Hospital Veterinário da Universidade Federal do Rio Grande do Sul, um canino, da raça Bulldog, de oito anos, pesando 32 kg, o qual habitava em um sítio. Foi relatado que o animal havia sofrido um acidente ofídico há duas horas e recebeu soro antiofídico em doses não conhecidas, por via subcutânea, cerca de uma hora após o acidente. A cobra foi descrita como da espécie Bothrops alternatus. O exame clínico mostrou edema acentuado na região submandibular direita, hematoma e lesões com sangramento no ponto de inoculação no pescoço. O animal foi hospitalizado e foram realizados hemograma e exames bioquímicos (ALT, creatinina, ureia), os quais não tiveram alterações, além de testes de coagulação (TP, TTPa), os quais se mostraram incoaguláveis (acima de 30s).(AU)