Study on the obtaining of Tityus trivittatus venom in Argentina.

Envenomation by scorpions of the genus Tityus is an important public health problem in Argentina, involving near 8000 stings and 2 deaths each year. Treatment for envenomation is the use of specific antivenom and intensive hospital care. Antivenom is produced by the Ministry of Health and freely distributed throughout the country. For antivenom production it is necessary to collect scorpion venom, which is a difficult task because although scorpions can be found in Argentina, they are less abundant than in warmer latitudes. For this reason venom collection constitutes a bottleneck for antivenom production. Although in Argentina several species of Tityus can be found, most of the accidents are caused by Tityus trivittatus, and the venom of this scorpion has historically been the venom used for antivenom production. We analyzed retrospectively 26 pools of telson homogenates (6964 telsons) and 37 pools of milked venom obtained by electrical stimulation (equivalent to 6841 milkings). Lethal potencies of samples from different provinces were very similar, although venom from scorpions of Buenos Aires city showed the lowest potency. The venom obtained by milking (median LD50 12.3 µg), provided batches containing LD50s more potent when compared with the venom obtained from telson homogenates (p < 0.0001). Many batches of telson homogenates (30%) showed lower potencies than acceptable for antivenom production and control. In addition to the study of the venom yield, the records of immunization of horses, the potency of the batches and the protein content of each batch of anti-scorpion antivenom produced were analyzed, comparing those produced using milked venom with those using telson homogenates as immunogens. Batches produced using milked venom required a shorter period of immunization (p < 0.0001), rendered higher neutralizing titers (p 0.0350) and possessed lower protein content (p 0.0092). Results clearly showed that the milking of scorpions is a more efficient tool to obtain venom for antivenom production in comparison to the use of telson homogenates.

Antivenom against Crotalus durissus terrificus venom: Immunochemical reactivity and experimental neutralizing capacity.

The venom of Crotalus durissus terrificus produces a neurotoxic and myotoxic syndrome that can lead to the death. Specific antivenom is the only treatment to neutralize the toxicity of the venom and the precocity in applying the antivenom is crucial for the efficiency of the treatment. We studied the variation of the immunochemical reactivity and neutralizing capacity of the specific antivenom on this venom in pre-incubation and rescue experiments, at different times. ELISA titers increased with longer venom-antivenom incubation times (p < 0.05) nevertheless incubation times had no effect on the neutralizing capacity of the antivenom. The antivenom dose necessary to rescue mice injected with 1.5 MMD (minimal mortal dose) 30 min after venom inoculation was over ten folds the dose of antivenom theoretically required to neutralize the same dose of venom according values obtained from pre-incubation experiments. Results showed that the in vitro immunochemical reactivity is not directly related to the neutralizing capacity. These observations underline the need for a rapid antivenom administration. Although preincubation experiments in mice are a powerful tool for the validation of the potency of the antivenoms in the productive process, it is clear that the nominal neutralization of the antivenoms must not be considered as a "stoichiometric" value regarding the venom to be neutralized in case of natural envenomation and emphasize the need of realization of clinical trials in order to evaluate the adequate doses of antivenom to be therapeutically used.

Toxicity of the venom of Latrodectus (Araneae: Theridiidae) spiders from different regions of Argentina and neutralization by therapeutic antivenoms.

"Black widow" spiders belong to the genus Latrodectus and are one of the few spiders in the world whose bite can cause severe envenomation in humans and domestic animals. In Argentina, these spiders are distributed throughout the country and are responsible for the highest number of bites by spiders of toxicological sanitary interest. Here, we studied the toxicity and some biochemical and immunochemical characteristics of eighteen venom samples from Latrodectus spiders from eight different provinces of Argentina, and the neutralization of some of these samples by two therapeutic antivenoms used in the country for the treatment of envenomation and by a anti-Latrodectus antivenom prepared against the venom of Latrodectus mactans from Mexico. We observed important toxicity in all the samples studied and a variation in the toxicity of samples, even in those from the same region and province and even in the same Latrodectus species from the same region. The therapeutic antivenoms efficiently neutralized all the venoms studied.

Lethality and histopathological alterations caused by Phoneutria nigriventer spider venom from Argentina: Neutralization of lethality by experimental and therapeutic antivenoms.

Although the spiders of the genus Phoneutria cause envenomation and their presence has been described in several provinces of the north of Argentina, they are not as common as other spiders of sanitary importance. In the present work, we studied the toxicity of samples of venom of Phoneutria spiders from the provinces of Misiones (where severe envenomation and deaths by Phoneutria have been recorded) and Jujuy (where no deaths have been recorded and severe envenomations are not frequent). To this end, we assessed the lethal potency in mice and guinea pigs and the histopathological alterations caused by both venoms, as well as the neutralization by the commonly used therapeutic antivenom produced by the Butantan Institute in Brazil and by an experimental antivenom developed with venom of P. nigriventer from Misiones. There were no differences in the lethality of the venoms of spiders from both regions. Post mortem examination showed that the heart and lungs were the most affected organs, while important pulmonary edema was seen macroscopically. Histological analysis showed edema, atelectasis, emphysema and cardiac lesion in both experimental models. The antivenoms assayed showed good neutralization of the venoms in the two experimental models. Despite the different geographic origins, the venoms showed similar toxicity and both the experimental antivenom and therapeutic antivenmos were able to neutralize the venoms of Argentinean P. nigriventer.

Venom yield and its relationship with body size and fang separation of pit vipers from Argentina.

The amount of venom that a snake can inject is related to its body size. The body size is related to head size and to the distance between fangs. To correlate snake body size, distance between fangs and distance between puncture wounds with the venom yield (and consequently with the venom dose potentially injected in a single snakebite), we studied these variables in two species of public health importance in South America, Bothrops (Rhinocerophis) alternatus, and Crotalus durissus terrificus. In all cases a positive correlation was observed between body length, fang separation distance, distance between puncture wounds and venom yield, with a regression coefficient over 0.5 for Bothrops alternatus and over 0.6 for Crotalus durissus terrificus in all cases, being the relation distance between punctures wounds and venom yield of 0.54 and 0.69 respectively. The difference between fang separation and puncture separation was never greater than 30%, with a mean difference around 13%. The strong relationships between body size, fang separation and venom yield may be useful for planning potential venom production in serpentariums. In addition, because puncture mark separation gives an approximate idea of the size of the snake, this provides a rough idea of the size of the snake that produced a bite and the potential amount of venom that could have been injected.

Identificación de los escorpiones de importancia médica en la Argentina / Identification of scorpions with medical importance in Argentina

. Los escorpiones de importancia médica en el mundo pertenecen mayoritariamente a la familia Buthidae. En Argentina, los miembros de esta familia no son numerosos, siendo mucho mayor el número de especies de la familia Bothriuridae, cuyos miembros hasta el momento no revisten importancia médica. Dentro de los tres géneros de Buthidae que viven en el país, el género Tityus es el que posee especies cuya picadura e inoculación de veneno puede provocar accidentes severos. En general, la diferenciación entre Tityus trivittatus, el escorpión de mayor importancia médica de Argentina, respecto a otros escorpiones, es fácil en las provincias de la zona central del país. Sin embargo, las reglas a utilizar para la diferenciación entre escorpiones de importancia médica de aquellos que no la tienen, no son similares en todo el país. En esta actualización, se brindan elementos sencillos y claros para que el personal de salud pueda diferenciar entre las especies de Tityus de importancia médica de los escorpiones de la familia Bothriuridae, ampliamente distribuidos en la superficie del país. De esta forma, se espera que el diag­nóstico acertado sobre la especie de escorpión pueda mejorar la atención en el caso de la picadura por animales potencialmente peligrosos. Por otro lado, permitirá diferenciar a los escorpiones sin importancia médica lo que redundará en la mejor utilización de los recursos disponibles en los niveles de atención primaria, ante la emergencia que representa una picadura de alacrán.

The scorpions of medical importance in the world mostly belong to Buthidae family. In Argentina, members of this family are not numerous, being much most numerous the species from the Bothriuridae family, whose members so far are of no medical importance. Within the three genera of Buthidae that inhabit in the country, only species from Tityus genus can cause severe ac­cidents. The differentiation between the scorpions of highest medical importance in Argentina (Tityus trivittatus), regarding other scorpions, is easy in the provinces of the central region. However, the rules used for the differentiation of medically important scorpions regarding those not dangerous, is not similar through the country. In this update, we provide simple and clear tools for the health personnel to differentiate between species of Tityus with medical importance and the scorpions from Bothriuridae fam­ily, widely distributed in the country. Thus it is expected that the accurate diagnosis of the species of scorpions may improve the attention in the case of the sting of potentially dangerous scorpions. On the other hand, the differentiation of the not dangerous scorpions from those with medical importance will result in the best use of the available resources in primary care level, consider­ing the emergency that represents a scorpion sting.

Identificación de los escorpiones de importancia médica en la Argentina / Identification of scorpions with medical importance in Argentina

. Los escorpiones de importancia médica en el mundo pertenecen mayoritariamente a la familia Buthidae. En Argentina, los miembros de esta familia no son numerosos, siendo mucho mayor el número de especies de la familia Bothriuridae, cuyos miembros hasta el momento no revisten importancia médica. Dentro de los tres géneros de Buthidae que viven en el país, el género Tityus es el que posee especies cuya picadura e inoculación de veneno puede provocar accidentes severos. En general, la diferenciación entre Tityus trivittatus, el escorpión de mayor importancia médica de Argentina, respecto a otros escorpiones, es fácil en las provincias de la zona central del país. Sin embargo, las reglas a utilizar para la diferenciación entre escorpiones de importancia médica de aquellos que no la tienen, no son similares en todo el país. En esta actualización, se brindan elementos sencillos y claros para que el personal de salud pueda diferenciar entre las especies de Tityus de importancia médica de los escorpiones de la familia Bothriuridae, ampliamente distribuidos en la superficie del país. De esta forma, se espera que el diag¡nóstico acertado sobre la especie de escorpión pueda mejorar la atención en el caso de la picadura por animales potencialmente peligrosos. Por otro lado, permitirá diferenciar a los escorpiones sin importancia médica lo que redundará en la mejor utilización de los recursos disponibles en los niveles de atención primaria, ante la emergencia que representa una picadura de alacrán.(AU)

The scorpions of medical importance in the world mostly belong to Buthidae family. In Argentina, members of this family are not numerous, being much most numerous the species from the Bothriuridae family, whose members so far are of no medical importance. Within the three genera of Buthidae that inhabit in the country, only species from Tityus genus can cause severe ac¡cidents. The differentiation between the scorpions of highest medical importance in Argentina (Tityus trivittatus), regarding other scorpions, is easy in the provinces of the central region. However, the rules used for the differentiation of medically important scorpions regarding those not dangerous, is not similar through the country. In this update, we provide simple and clear tools for the health personnel to differentiate between species of Tityus with medical importance and the scorpions from Bothriuridae fam¡ily, widely distributed in the country. Thus it is expected that the accurate diagnosis of the species of scorpions may improve the attention in the case of the sting of potentially dangerous scorpions. On the other hand, the differentiation of the not dangerous scorpions from those with medical importance will result in the best use of the available resources in primary care level, consider¡ing the emergency that represents a scorpion sting.(AU)

Intraspecific differences in the immunochemical reactivity and neutralization of venom from Argentinean Bothrops (Rhinocerophis) alternatus by specific experimental antivenoms.

The venoms of Bothrops (Rhinocerophis) alternatus (B.a.) from different regions of Argentina have shown biochemical, toxicological and immunological variations. Considering these variations, we produced nine experimental antisera (rabbit, IgG) against venoms from snakes of nine different regions and a pool of venom, comprised of equal amounts of venoms from each region. The immunologic studies (ELISA, Westernblot) showed significant cross reactivity among all regional antivenoms with all regional venoms, with no significant differences regarding the specificity of the immunogens used for the production of antivenom. Neutralization of hemorrhage was variable (although all the antivenoms neutralized this activity in all venoms) and the neutralization of coagulant and phospholipase activities were evident in all cases. Some antivenoms neutralized toxic activities that were absent or very low in the venoms used as immunogen, on other non-homologous venoms (e.g. thrombin like activity). Despite the different toxic potencies of regional venoms, antivenoms developed using venoms of snakes from a particular region showed high immunochemical reactivity and cross-neutralizing capacity on snake venoms from different and distant regions, in occasions over those of the homologous antivenoms. These findings could be used to improve the generation of pools of venoms for the production of antivenoms.

Neutralization of Bothrops alternatus regional venom pools and individual venoms by antivenom: a systematic comparison.

In this study we report that variation in lethality, hemorrhagic potency and procoagulation between individual samples of Bothrops alternatus venom from a single region, and variation between regional pools at the national level are comparable in range. Furthermore, the range of relative neutralization potencies of individual venoms within a region overlaps, and sometimes exceeds, the range of neutralization of regional venom pools throughout the country. Thus, the potency of neutralization of a national venom pool is poorly predictive of the potencies of neutralization of its constituent regional venom pools and, furthermore, the potency of neutralization of a regional venom pool is poorly predictive of the potencies of neutralization of its individual venom constituents. The efficiencies of neutralization of each of these effects (lethality, hemorrhage and procoagulation) were not significantly related to each other and did not correlate to the corresponding toxic potency of each venom or venom pool. Some implications of these findings are discussed in the context of the distinction between experimental quantitation of antivenom potency and the amount of antivenom that might be actually required to successfully treat two apparently comparable B. alternatus envenomations.