A low-cost method to test cytotoxic effects of Crotalus vegrandis (Serpentes: Viperidae) venom on kidney cell cultures

The pathogenesis of the renal lesion upon envenomation by snakebite has been related to myolysis, hemolysis, hypotension and/or direct venom nephrotoxicity caused by the venom. Both primary and continuous cell culture systems provide an in vitro alternative for quantitative evaluation of the toxicity of snake venoms. Crude Crotalus vegrandis venom was fractionated by molecular exclusion chromatography. The toxicity of C. vegrandis crude venom, hemorrhagic, and neurotoxic fractions were evaluated on mouse primary renal cells and a continuous cell line of Vero cells maintained in vitro. Cells were isolated from murine renal cortex and were grown in 96 well plates with Dulbecco's Modified Essential Medium (DMEM) and challenged with crude and venom fractions. The murine renal cortex cells exhibited epithelial morphology and the majority showed smooth muscle actin determined by immune-staining. The cytotoxicity was evaluated by the tetrazolium colorimetric method. Cell viability was less for crude venom, followed by the hemorrhagic and neurotoxic fractions with a CT50 of 4.93, 18.41 and 50.22 µg/mL, respectively. The Vero cell cultures seemed to be more sensitive with a CT50 of 2.9 and 1.4 µg/mL for crude venom and the hemorrhagic peak, respectively. The results of this study show the potential of using cell culture system to evaluate venom toxicity.

Aspectos da biologia reprodutiva de Bothrops jararaca em cativeiro (Serpentes, Viperidae)

The reproduction of Bothrops jararaca (Wied, 1824) in captivity in ordened to determine the intercourse period, births, number of neonates, proportion of males and females per litter were performed between 1989 and 1999. Fifty eight females were observed, 16 intercourses in captivity, 45 females arrived already fertilized from nature and 53 litters, resulting on the birth of 426 live neonates, 67 infertile egg masses, 18 stillborn neonates and 4 abnormal neonates. The intercourse period was from february to december and the births happened between february and may. From the two gestacional periods observed, the shortest one was 152 days and the longest 239. The average amount of live neonates per litter was 8,04. Among 323 neonates, 47,68% were males and 52,32% were females. During the first year of life, 75,71% of the females and 71,54% of the males died, and 2,31% of the females and 0,81% of the males reached the fifth year of life.

Ocorrência de híbridos não naturais entre Bothrops jararaca e B. neuwiedi (Serpentes, Viperidae)

In May 1987, a female of Bothrops jararaca (Wied, 1824), from Carazinho, Rio Grande do Sul (RS), Brazil, was placed in the same vivarium with a male of Bothrops neuwiedi Wagler, 1824 coming from Guaíba, RS. There, they stayed for aproximately ten months. In March 1988, it was observed a delivery of five live and two still born, among them six presented morphologic characteristics of B. neuwiedi and one of B. jararaca. After the female died, in April 1988, through necropsy, two fetusus were found, one near the cloaca and, both identified as B. neuwiedi. The morphologic analysis and the origin of the progenitors suggest the hypothesis that the litter was resulted of cross-breeding.

Variação anual do sistema reprodutor de fêmeas de Bothrops jararaca (Serpentes, Viperidae)

The morphological changes of the ovary and oviduct of 238 tropical snakes Bothrops jararaca (Wied, 1824) were determined. The ovarian mass presented a remarkable decrease in October, after ovulation, staying in low levels from November to March, during the gestational period. From April to September, it increased because of the ovarian follicles maturation. A gradual increase in oviduct weight was observed from October to March due to stages of embryonic development. A significant difference was observed between right and left ovary weight, and oviduct length, independently of the months considered.

Capacidade reprodutiva de Bothrops jararaca (Serpentes, Viperidae)

The relationship between female body size and egg number was examined in the snake Bothrops jararaca (Wied, 1924). Some authors have demonstrated that the estimated clutch size may be affected by the inclusion or exclusion of dead neonates or non-developed eggs. The reproductive system of 254 mature females newly arrived from nature was examined and 29.55% (n=75) had eggs in the oviducts in different stages of embryonic development. The presence of atresic eggs was observed in 34.5% of those females. A significant difference was observed in the number of eggs in the right and left oviducts (t=9.48, P<0.001), but not in the number of atresic eggs (t=1.110, P=0.278). The correlation of female body size and egg number was greater when all, the normal and atresic eggs were considered (r=0.63, P<0.001). The exclusion of atresic eggs from the clutch size decreased slightly this correlation (r=0.54, P<0.001).

Variação morfométrica em Bothropoides jararaca (Serpentes, Viperidae) no Rio Grande do Sul

Apesar de machos e fêmeas de serpentes nascerem com o mesmo tamanho, as taxas de crescimento e a idade da maturação sexual podem ser diferentes, determinando dimorfismo sexual em estágios posteriores da vida. Avaliamos a ocorrência de variação morfométrica sexual e ontogenética em Bothropoides jararaca (Wied, 1824), explorando as relações entre tamanho corporal e amadurecimento sexual através de 14 variáveis morfométricas. Foram analisados 142 espécimes provenientes do estado do Rio Grande do Sul, sul do Brasil. Os dados morfométricos - comprimento da cabeça, rostro-cloacal, da cauda, comprimento total; largura da cabeça, ocular, nasal, loreal, da cauda; distância ocular-nasal, ocular-loreal, loreal-nasal, ventral-sinfisal e rostral-labial - e comprimento/diâmetro dos folículos ovarianos, foram tomados em milímetros, através de régua simples e paquímetro analógico de precisão 0,05 mm. A determinação sexual foi realizada por inspeção das gônadas. A classificação etária foi associada à maturidade sexual. Para as análises estatísticas foram utilizadas análises de variância (ANOVA) com teste de Tukey post hoc, regressão linear e análise discriminante canônica (ADC). A maioria das medidas indicou dimorfismo sexual (ANOVA, P<0,05) apenas em adultos (Tukey, P<0,05). As análises de regressão mostram que o comprimento rostro-cloacal explica o comportamento das demais variáveis (P<0,001) e que em todas as medidas as fêmeas crescem mais que os machos. A ADC foi exitosa em separar as classes sexuais e etárias, apresentando significado biológico, considerando 79,2% dos casos como corretamente classificados.

Occurrence of Cryptosporidium (Apicomplexa, Cryptosporidiidae) in Crotalus durissus terrificus (Serpentes, Viperidae) in Brazil

The objective of the present study was to investigate the prevalence of Cryptosporidium (Apicomplexa, Cryptosporidiidae) in the snake Crotalus durissus terrificus (Serpentes, Viperidae). Fifty animals were evaluated for the presence of oocysts of Cryptosporidium sp. at the time of arrival and 30 and 60 days later. Intestinal washings with saline solution (1% body weight), fecal samples, and organ scrapings were collected during the study. Oocysts were concentrated by an ether-phosphate-buffered saline sedimentation technique and then separated by a density gradient centrifugation technique. Smears were made with the sediment and submitted to modified acid-fast and auramine-rhodamine staining. Cryptosporidium-positive smears were used as controls for the experimental findings. The overall prevalence of Cryptosporidium sp. oocysts was 14%. Among the positive snakes, oocysts were detected only in the intestinal washing in two specimens, only in the feces in four specimens, and in both materials at least once in one specimen. The positive snakes were predominantly from Santa Maria da Serra city State of São Paulo (57.1%). We also observed that all of the examinations that presented positive results were obtained at least 27 days after the capture of the animals.

Occurrence of Cryptosporidium (Apicomplexa, Cryptosporidiidae) in Crotalus durissus terrificus (Serpentes, Viperidae) in Brazil

The objective of the present study was to investigate the prevalence of Cryptosporidium (Apicomplexa, Cryptosporidiidae) in the snake Crotalus durissus terrificus (Serpentes, Viperidae). Fifty animals were evaluated for the presence of oocysts of Cryptosporidium sp. at the time of arrival and 30 and 60 days later. Intestinal washings with saline solution (1% body weight), fecal samples, and organ scrapings were collected during the study. Oocysts were concentrated by an ether-phosphate-buffered saline sedimentation technique and then separated by a density gradient centrifugation technique. Smears were made with the sediment and submitted to modified acid-fast and auramine-rhodamine staining. Cryptosporidium-positive smears were used as controls for the experimental findings. The overall prevalence of Cryptosporidium sp. oocysts was 14%. Among the positive snakes, oocysts were detected only in the intestinal washing in two specimens, only in the feces in four specimens, and in both materials at least once in one specimen. The positive snakes were predominantly from Santa Maria da Serra city State of São Paulo (57.1%). We also observed that all of the examinations that presented positive results were obtained at least 27 days after the capture of the animals.

The occurrence of Crepidobothrium sp. (Cestoda, Proteocephalidae) in Bothrops moojeni (Hoge) (Serpentes, Viperidae)

The occurrence of Crepidobothrium sp.(Cestoda, Proteocephalidae) in the intestine of Bothrops moojeni (Hoge,1965)(Serpentes, Viperidae) is reported. The host snake was rescued from the fauna in Porto Primavera dam, Mato Grosso do Sul State, Brazil. The snake died in captivity on July 13,1999. At necropsy, 28 tapeworms were found in the snake intestine. The analysis of specimens morphology allowed the conclusion that they belong to the Crepidobothrium (Monticelli, 1900) genus. It was not possible to determine the Crepidobothrium species due to the lack of the gravid proglottids. This is the first report of B. moojeni as a host of cestodes.

Morphological, morphometric, and molecular characterization of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from naturally infected Caudisona durissa terrifica (Serpentes, Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Description of gamontogonic and sporogonic stages of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from Caudisoma durissa terrifica (Serpentes, Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Megaselia scalaris (Diptera : Phoridae) causing myiasis in Crotalus durissus terrificus (Serpentes : Viperidae) in Brazil

We describe a case of myiasis in Crotalus durissus terrificus (Laurenti) caused by Megaselia scalaris (Loew). The snake was Found in Anhembi, São Paulo, Brazil, with a lesion measuring 25 mm in diameter where the larvae of M. scalaris had penetrated the ribs. The opportunistic behavior of the larvae of M. scalaris is discussed.

Meal size and specific dynamic action in the rattlesnake Crotalus durissus (Serpentes : Viperidae)

We studied the effect of meal size on specific dynamic action (SDA) in the South American rattlesnake Crotalus durissus, by measuring oxygen consumption rates (VO2) prior to and after the ingestion of meals ranging from 10-50% of snake's body mass. Regardless of meal size, variation in VO2 with time during digestion demonstrated the same general pattern. Oxygen consumption rates peaked between 15 and 33 h post-feeding, at values 3.7-7.3 times those prior to feeding. Snakes, while digesting meals of 30% and 50% of their body mass, experienced VO2 that exceeded rates measured during forced activity. Following peaks in VO2, rates returned to prefeeding values within 62-170 h post-feeding. Post-prandial peak in VO2 and the duration of the metabolic response to feeding increased with meal size. Digestion is an energetically demanding activity for C. durissus, with an estimated cost equaling 12-18% of the ingested assimilated energy.

Diet of the rattlesnake Crotalus durissus in southeastern Brazil (Serpentes, Viperidae)

Gut contents of 633 live rattlesnakes from southeastern Brazil received at the Instituto Butantanitute, SP, Brazil between 1993 and 1995 were studied. The snakes were weighed, measured and sexed. Two hundred and fifty-nine rattlesnakes had stomach and/or intestinal contents. Prey size was estimated by comparison of prey items with specimens from museum collections. Rodents and small marsupials were the main prey eaten by the rattlesnakes, and only 1% of the items were found in the stomach, whereas 41% of the individuals in the sample had feces in the intestine. There was low correlation between size of snake and prey size. No seasonal difference in frequency was found between fed and not fed males, but the occurrence of fed females was significantly lower during summer than winter months (28.9% and 51.8%, respectively). Fed newborn rattlesnakes had the lowest frequency, and also fed on rodents.

Postprandial thermogenesis in Bothrops moojeni (Serpentes: Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)