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Pdf/A07v18n4.Pdf 35 RESEARCH ARTICLE Effectiveness of Lonomia antivenom in recovery from the coagulopathy induced by Lonomia orientoandensis and Lonomia casanarensis caterpillars in rats Ida S. Sano-Martins1*, Camila GonzaÂlez2, Isabelle Valle Anjos1, Juana DõÂaz2, Luis Roberto C. GoncËalves1 a1111111111 1 LaboratoÂrio de Fisiopatologia, Instituto Butantan, São Paulo±SP, Brazil, 2 Centro de Investigaciones en a1111111111 MicrobiologõÂa y ParasitologõÂa Tropical (CIMPAT), Facultad de Ciencias, Universidad de los Andes, BogotaÂ, a1111111111 Colombia a1111111111 a1111111111 * [email protected], [email protected] Abstract OPEN ACCESS In South America, accidental contact with Lepidoptera larvae can produce a diversity of Citation: Sano-Martins IS, GonzaÂlez C, Anjos IV, reactions that vary from dermatological problems to severe hemorrhagic syndromes, such DõÂaz J, GoncËalves LRC (2018) Effectiveness of as those caused by contact with caterpillars of the genus Lonomia (Saturniidae). Lonomia Lonomia antivenom in recovery from the venom can alter the hemostatic system and lead to renal failure, internal and brain bleeding, coagulopathy induced by Lonomia orientoandensis and in severe cases, death. The only specific treatment available for these envenomations and Lonomia casanarensis caterpillars in rats. PLoS Negl Trop Dis 12(8): e0006721. https://doi. is the Lonomia Antivenom (LAV) produced by the Butantan Institute, in Brazil, using an org/10.1371/journal.pntd.0006721 extract of Lonomia obliqua scoli as the antigen. LAV has been used to treat exposure to Editor: Jean-Philippe Chippaux, Institut de other Lonomia species across South America. However, no experimental studies have Recherche pour le DeÂveloppement, BENIN been performed to test the efficacy of LAV in neutralizing the venom of species other than L. Received: May 17, 2018 obliqua found in Southern Brazil. In this study, we tested the effectiveness of LAV in revers- ing the hemostatic disturbances induced by injecting Lonomia casanarensis (Lca) and Lono- Accepted: July 26, 2018 mia orientoandensis (Lor) scolus extracts into rats and compared the effects to the case of Published: August 16, 2018 L. obliqua (Lob) scolus extract-induced envenomation. Lca and Lor caterpillars were col- Copyright: © 2018 Sano-Martins et al. This is an lected in Colombia, and some of them were reared to adults for identification. The Minimum open access article distributed under the terms of Defibrinating Doses (MDD) of Lca and Lor were estimated. Rats were injected (i.d.) with a the Creative Commons Attribution License, which permits unrestricted use, distribution, and dose of 3 MDD per rat of each scolus extract and treated (i.v.) with 1.5 mL of LAV or 1.5 mL reproduction in any medium, provided the original of saline. Twenty-four hours after the treatment, the fibrinogen levels and platelet counts author and source are credited. had recovered to the hemostatic levels in the groups treated with LAV. The groups treated Data Availability Statement: All relevant data are with the saline solution had fibrinogen levels and platelet counts at non-hemostatic levels. within the paper. Thromboelastometric analyses confirmed these results. In conclusion, the results showed Funding: This work was funded by Vicerectoria de that LAV is effective at neutralizing the envenomation induced by Lca and Lor spine extracts Investigaciones, Faculdad de Ciencias, Universidad in rats and restoring hemostasis. de Los Andes, Convocatoria 2016-2. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. PLOS Neglected Tropical Diseases | https://doi.org/10.1371/journal.pntd.0006721 August 16, 2018 1 / 16 Lonomia antivenom effectiveness against two new Lonomia species Author summary A specific antivenom therapy is recommended for treating envenomation caused by ven- omous animals, such as snakes, spiders, scorpions, and Lonomia caterpillars. Studies with snake bites have shown that antivenoms are effective for treating the envenomation caused by snakes of the same genus, even when the antigen composition used to generate the antivenom did not include the specific venom. The Lonomia antivenom produced by the Butantan Institute to treat envenomations caused by contact with Lonomia caterpillars only uses the extract of Lonomia obliqua scoli as the antigen. However, there are reports of other species of Lonomia in South America that demonstrate biological activities that are different from those observed in response to L. obliqua venom, but which cause similar symptoms in human envenomation. No information is available regarding the neutraliz- ing capacity of the Lonomia antivenom in these other species. This study showed that the Lonomia antivenom efficiently reversed the hemostatic disorders induced in rats by injec- tion of extracts of the scoli of L. casanarensis and L. orientoandensis from Colombia. These results support the possibility of using this antivenom for treating envenomations caused by other Lonomia species in humans. Introduction Several families within the Lepidoptera (moths and butterflies) produce harmful venoms. One group, the family Saturniidae, includes a genus, Lonomia (Saturniidae: Hemileucinae), of par- ticular public health concern [1±3]. As a part of their defense mechanisms, Lonomia caterpillars are gregarious and form groups of over fifty individuals, have cryptic colors, and feed at night on host plant leaves [4]. The spines of Lonomia obliqua Walker, 1855 caterpillars are associated with a gland that is a single large cell in the subapical region [5]. Venom is released when the spine breaks after touching or penetrating a rigid surface, such as the human skin, allowing venom absorption [4±6]. Symptoms of the envenomation that results from contacting Lonomia caterpillars can vary from intense local pain or a burning sensation at the site of contact, to renal failure and intra- cerebral and intraperitoneal hemorrhages that can lead to death [7±10]. Several studies of the composition of the scolus extract and toxicity have been conducted in Brazil and Venezuela. Regarding their epidemiological importance, two species are involved in accidents: Lonomia achelous Cramer, 1777 and L. obliqua [9,11±13]. Lonomia obliqua is distributed in the southern region of South America, including Brazil, Uruguay, Paraguay, and northern Argentina, while L. achelous is present in the northern part of South America in Brazil, Ecuador, Peru, Venezuela, Guyana, and Colombia [14]. The first records of envenomation by Lonomia caterpillars in South America came from Ven- ezuela in 1967 and involved L. achelous caterpillars [8,15,16]. Brazil and Venezuela have reported high numbers of cases of contact with L. obliqua and L. achelous [8,9], and in recent years, Argen- tina [17±19] and Peru [20] have also indicated an increasing number of accidents and fatalities. In Colombia, most cases are recorded in the Casanare Department, but there is no available information regarding the real number of accidents in Colombia or the species that were involved. The official records noted 32 cases between 2000 and 2010, with two deaths [4,21,22]. Since 1994, the recommended treatment of envenomation from contacting Lonomia cater- pillars in Brazil is the use of Lonomia antivenom (LAV), which is produced using an extract of the L. obliqua scoli as the antigen [23]. In Venezuela, the treatment is based on the use of antifi- brinolytics, such as aprotinin, or the transfusion of fibrinogen or cryoprecipitate [24]. In PLOS Neglected Tropical Diseases | https://doi.org/10.1371/journal.pntd.0006721 August 16, 2018 2 / 16 Lonomia antivenom effectiveness against two new Lonomia species Colombia, treatment LAV has been successful, although the efficacy of this antivenom for Colombian Lonomia species and treatment protocols have not been yet established [21,22]. In this context, this study aimed to test the effectiveness of the Lonomia antivenom pro- duced in Brazil by the Butantan Institute for treating rats envenomed by an injection of scolus extracts from two caterpillar species collected in Colombia, Lonomia casanarensis Brechliin, 2017 and Lonomia orientoandensis Brechliin, 2011. Materials and methods Ethics statements and animals All procedures involving the use of animals were performed under the resolutions of the National Council for the Control of Animal Experimentation (Conselho Nacional de Controle de ExperimentacËão AnimalÐCONCEA) and approved by the Ethics Committee of Animal Use the Butantan Institute (protocol 1353/14 and 63000505/17). Male Wistar rats that weighed 250±300 g were supplied by the Central Animal Laboratory of the Instituto Butantan. Rats were housed in plastic cages, within a room in which the temperature was controlled to between 22 and 23ÊC. Rats were exposed to a 12:12 h light:dark cycle and were allowed free access to food and water. Scolus extracts and Lonomia antivenom Colonies of L. orientoandensis (Lor) and L. casanarensis (Lca) in the 5th and 6th instars were col- lected in the Department of Casanare, Colombia. Lca caterpillars were collected in the munici- pality of Tauramena (4.90773ÊS, -72.5437ÊW) and Lor caterpillars in the municipality of NunchõÂa (5.64629ÊS, -72.2011ÊW) (Fig 1). The colonies were transported to Bogota in plastic boxes supplied with host plant leaves. The extraction of the venom was carried out in the Centro de MicrobiologõÂa y ParasitologõÂa
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