SPECIAL ARTICLE Scorpion maintenance in captivity for venom extraction in Costa Rica Emanuel Brenes1* & Aarón Gómez2 1. Escuela de Ciencias Biológicas, Universidad Nacional, 3000 Heredia, Costa Rica; [email protected] 2. Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, 2060 San José, Costa Rica; [email protected] Received 09-IX-2015. Corrected 03-III-2016. Accepted 04-IV-2016. Abstract: Approximately 2 000 scorpion species can be found around the world; although few species are considered “harmful” to human beings, a high number of scorpionism cases are reported all over the world. The elaboration of anti-scorpion sera requires the establishment of an animal collection maintained in captivity for venom extraction purposes. The Clodomiro Picado Institute (ICP, for its acronym in Spanish), poses a vast tra- jectory in manufacturing snakebite antivenoms, and starts a scorpion collection in 2005 for this purpose. In total, 2 043 scorpions were classified in 11 species and collected during a seven-year period using a black-light flash- light and an intensive seeking methodology. The scorpions were collected from several localities of the Pacific and the Caribbean versants of Costa Rica. The venom extraction was performed by applying electrostimulation; the collected venom was characterized by total protein content in addition to median lethal doses. Centruroides bicolor showed higher amounts of venom yield, total protein content and more lethal dose, all of which were cor- related with its body mass. The techniques used to keep scorpions in captivity allowed the animals to live several years. Longevity analysis showed significant differences among scorpion genera (H= 353.80; df= 3; P < 0.0001); moreover, the genus Didymocentrus lived longer with an average of 4.46 years. One key factor of its longevity was that it did not go through venom extraction processes. Additionally, a high survival rate of Tityus pachyurus born in captivity, compared to other species within the same genus, was observed (H= 94.32; df= 3; P < 0.0001). This characteristic should be taken into consideration, when programs of reproduction in captivity are designed. In conclusion, the maintenance of a scorpion collection was efficient for venom extraction purposes and a longer life expectancy of the animals. Moreover, there is a scarcity on publications regarding scorpion maintenance in captivity for venom extraction purposes; therefore, a deeper research in aspects such as reproduction, death causes and feeding behaviors is required. Rev. Biol. Trop. 64 (3): 1019-1027. Epub 2016 September 01. Key words: Costa Rican scorpions, maintaining scorpions, captive scorpions, scorpion venom, venom extraction. Approximately 2 000 species of scorpions beings, where 19 species belong to the family can be found around the world, and are classi- Buthidae, and the genus Centruroides and Tity- fied in 15 families (Fet & Soleglad, 2005; Rein, us, found in America (Chippaux & Goyffon, 2013). Very few of these animals are consid- 2008). The Centruroides genus is distributed ered “dangerous” for human beings, and the from the North part of the American continent main scorpionism cases have been reported for to Central America, and the Tityus genus is regions such as: Australia (Isbister, Volschenk, found from Central America through South & Seymour, 2004), Asia (Dehghani, Vazirian- America (Borges et al., 2012). zadeh, Nasrabadi, & Moravvej, 2010), Africa The highest annual incidence of scorpion (Laïd et al., 2012), and America: especially stings in America occurs in Mexico, with 2 050 Latin America (Borges, Miranda, & Pascale, scorpion stings by 100 000 inhabitants (Chip- 2012). An approximate number of 30 species paux & Goyffon, 2008). Other Latin American of scorpions are potentially harmful for human countries have lower scorpion sting incidence, Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 64 (3): 1019-1027, September 2016 1019 for instance Brazil, Venezuela and Colombia a public health impact is fundamental. The with values of 142, 40, 5 scorpion sting by elaboration of scorpion antivenoms requires 100 000 inhabitants, respectively. It is worth the establishment of a collection in order to mentioning that some of these scorpion stings maintain the animals and for venom extrac- end up in fatalities, especially in children tion. A previous effort was made by Bucherl (Chippaux & Goyffon, 2008). Due to the scor- of the Butantán Institute, that described the pionism occurrence in Latin America, the need techniques for keeping a live collection of for an antivenom elaboration has emerged. scorpions in laboratory conditions for venom Thus, several commercial antivenoms are pro- extraction purposes (Whittemore, Keegan, duced in different countries such as Brazil Fitzgerald, Bryant, & Flanigan, 1963). Thus, (Candido & Lucas, 2004), Mexico (Chippaux the aim of this work was to report the experi- & Goyffon, 2008) and Venezuela (Poggioli De ences gathered in keeping a live collection of Scannone, 1996). Costa Rican scorpions, for venom extraction, In Central America the studies on scor- at the Clodomiro Picado Institute. pions are scarce, probably due to the reduced amount of species that inhabit these countries. GENERAL PROCEDURES AND Costa Rica reports 14 species within seven gen- PRELIMINARY FINDINGS era and four families; with the genus Centruroi- des and Tityus presenting medical importance An emergence and importance of the scor- due to their envenomation features. Moreover, pionism in Latin America has increased in the these two genera represent an emerging poten- last few years, especially in countries such as tial problem to the public health system, for Costa Rica, where hundreds of scorpion stings Costa Rica and the other Central American have been reported, including one fatality in countries (Leveridge, 2000). 1995 (Leveridge, 2000). For this reason, a In Costa Rica, the Clodomiro Picado Insti- group of scientists of the Clodomiro Picado tute has a remarkable work on research and Institute, started a research project on the elaboration of snakebite antivenoms (Rojas, characterization and, eventually, of a pilot anti- Jiménez, & Gutiérrez, 1993); however, the scorpion serum batch elaboration. scorpionism was not been attended until now. Furthermore, the National Center of Intoxi- Scorpions capture field trips: Sched- cation has reported an increased number of uled field trips were conducted from 2005 scorpion stings in the country since the 90’s through 2011 to the following locations: Santa decade: only 24 cases for 1992, 206 cases in Rosa National Park, Palo Verde National Park, 1998, and 405 cases in 1999, practically a Quepos, Playa Piro, Cabo Blanco and Carara 100 % increase (Leveridge, 2000). The current in the Pacific versant; Hitoy Cerere National problem is that there is no proper antivenom Park, Chiroles, La Selva Biological Station, for its treatment, and the gap between the scor- Siquirres, Guayacan, Barbilla National Park pionism stings problem and an effective treat- and La Gamba in the Caribbean versant of ment became evident. Considering the scorpion Costa Rica (Fig. 1). Additionally, some scorpi- stings relevance in public health and the poten- ons were provided by particulars. The scorpions tial harm that it may cause, the Clodomiro were collected using an ultraviolet flashlight Picado Institute decided to study the toxins of and an intensive seeking strategy; captured the major scorpion species inducing pathologic scorpions were transported to the Clodomiro effects, as well as the establishment of a live Picado Institute located in Dulce Nombre de animal maintenance installation for venom Coronado, San José, Costa Rica; and were reg- extraction purposes. istered in the collection. The following species Nowadays, the establishment of collec- were collected during such field trips: Ananter- tions in captivity of animals that may cause is platnicky (Lourenço, 1982), Didymocentrus 1020 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 64 (3): 1019-1027, September 2016 -86º0.000’ -84º0.000’ Nicaragua Caribbean Sea Species Ananteris platnicky Centruroides bicolor 10º0.000’ Centruroides koesteri Centruroides limbatus Centruroides edwarsii Didymocentrus krausi Tityus cerroazul Tityus championi Tityus dedoslargos Ocean Pacic Tityus ocelote Tityus pachyurus Panamá Fig. 1. Scorpion collecting sites in Costa Rica during field trips conducted by the Clodomiro Picado Institute. krausi (Francke, 1978), Centruroides edwardsii this genus in Costa Rica can tolerate alterations (Gervais, 1843), C. koesteri (Kraepelin, 1911), in the forest and are even well adapted to urban C. bicolor, C. limbatus, Tityus championi and environments (Víquez, 1999). On the other T. pachyurus (Pocock, 1898), T. cerroazul hand, C. limbatus is the most common spe- (Lourenço, 1986), T. dedoslargos and T. oce- cies in the Caribbean, and C. bicolor the most lote (Francke & Stockwell, 1987). common in the South Pacific (Víquez, 1999). On the other hand, 88 % of the animals Scorpions of the genus Tityus showed arboreal were collected by the Clodomiro Picado Insti- habits and can be found in unaltered to less tute staff during field trips. In Santa Rosa, altered forests (Borges et al., 2012); this made Guanacaste, 800 scorpions were captured dur- its observation and capture difficult, resulting ing several field trips. The most common spe- in less encounters. cies found in this locality was C. edwardsii, however, C. koesteri and D. krausi