title: Combined venomics, venom gland transcriptomics, bioactivities, and antivenomics of two Bothrops jararaca populations from geographic isolated regions within the Brazilian Atlantic rainforest.
abstract: Bothrops jararaca is a slender and semi-arboreal medically relevant pit viper species endemic to tropical and subtropical forests in southern Brazil, Paraguay, and northern Argentina (Misiones). Within its geographic range, it is often abundant and is an important cause of snakebite. Although no subspecies are currently recognized, geographic analyses have revealed the existence of two well-supported B. jararaca clades that diverged during the Pliocene ~3.8Mya and currently display a southeastern (SE) and a southern (S) Atlantic rainforest (Mata Atlântica) distribution. The spectrum, geographic variability, and ontogenetic changes of the venom proteomes of snakes from these two B. jararaca phylogroups were investigated applying a combined venom gland transcriptomic and venomic analysis. Comparisons of the venom proteomes and transcriptomes of B. jararaca from the SE and S geographic regions revealed notable interpopulational variability that may be due to the different levels of population-specific transcriptional regulation, including, in the case of the southern population, a marked ontogenetic venom compositional change involving the upregulation of the myotoxic PLA2 homolog, bothropstoxin-I. This population-specific marker can be used to estimate the proportion of venom from the southern population present in the B. jararaca venom pool used for the Brazilian soro antibotrópico (SAB) antivenom production. On the other hand, the southeastern population-specific D49-PLA2 molecules, BinTX-I and BinTX-II, lend support to the notion that the mainland ancestor of Bothrops insularis was originated within the same population that gave rise to the current SE B. jararaca phylogroup, and that this insular species endemic to Queimada Grande Island (Brazil) expresses a pedomorphic venom phenotype. Mirroring their compositional divergence, the two geographic B. jararaca venom pools showed distinct bioactivity profiles. However, the SAB antivenom manufactured in Vital Brazil Institute neutralized the lethal effect of both venoms to a similar extent. In addition, immobilized SAB antivenom immunocaptured most of the venom components of the venoms of both B. jararaca populations, but did not show immunoreactivity against vasoactive peptides. The Costa Rican bothropic-crotalic-lachesic (BCL) antivenom showed the same lack of reactivity against vasoactive peptides but, in addition, was less efficient immunocapturing PI- and PIII-SVMPs from the SE venom, and bothropstoxin-I, a CRISP molecule, and a D49-PLA2 from the venom of the southern B. jararaca phylogroup. The remarkable paraspecificity exhibited by the Brazilian and the Costa Rican antivenoms indicates large immunoreactive epitope conservation across the natural history of Bothrops, a genus that has its roots in the middle Miocene. This article is part of a Special Issue entitled: Omics Evolutionary Ecolog.
authors.affiliation: Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Laboratório de Hemostase e Venenos, Universidade Federal do Rio de Janeiro (UFRJ), Brazil; Instituto Nacional de Biologia Estrutural e Bioimagem, Rede Proteomica do Rio de Janeiro, Universidade Federal do Rio de Janeiro (UFRJ), Brazil; Laboratorio de Venómica Estructural y Funcional, Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain.
authors.forename: Larissa
authors.initials: L
authors.lastname: Gonçalves-Machado
authors.affiliation: Laboratorio de Venómica Estructural y Funcional, Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain. Electronic address: dpla@ibv.csic.es.
authors.forename: Davinia
authors.initials: D
authors.lastname: Pla
authors.affiliation: Laboratorio de Venómica Estructural y Funcional, Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain.
authors.forename: Libia
authors.initials: L
authors.lastname: Sanz
authors.affiliation: Departamento de Fisiologia e Farmacologia, Universidade Federal do Ceará (UFC), Fortaleza 60430-270, Brazil.
authors.forename: Roberta Jeane B
authors.initials: RJB
authors.lastname: Jorge
authors.affiliation: Fundação Zoobotânica do Rio Grande do Sul, Museu de Ciências Naturais, Núcleo Regional de Ofiologia de Porto Alegre, RS, Brazil.
authors.forename: Moema
authors.initials: M
authors.lastname: Leitão-De-Araújo
authors.affiliation: Fundação Zoobotânica do Rio Grande do Sul, Museu de Ciências Naturais, Núcleo Regional de Ofiologia de Porto Alegre, RS, Brazil.
authors.forename: Maria Lúcia M
authors.initials: MLM
authors.lastname: Alves
authors.affiliation: Laboratorio de Herpetologia, Departamento de Zoologia, Instituto de Biociências, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçãlves 9500, Agronomia, 91501-970 Porto Alegre, RS, Brazil.
authors.forename: Diego Janisch
authors.initials: DJ
authors.lastname: Alvares
authors.affiliation: Hygeia Biotecnologia Aplicada S.A., Fundação Bio-Rio, Rio de Janeiro, Brazil.
authors.forename: Joari
authors.initials: J
authors.lastname: De Miranda
authors.affiliation: Hygeia Biotecnologia Aplicada S.A., Fundação Bio-Rio, Rio de Janeiro, Brazil.
authors.forename: Jenifer
authors.initials: J
authors.lastname: Nowatzki
authors.affiliation: Laboratorio de Herpetologia, Instituto Butantan, Avenida Vital Brazil 1500, São Paulo 05503-900, SP, Brazil.
authors.forename: Karen
authors.initials: K
authors.lastname: de Morais-Zani
authors.affiliation: Laboratorio de Herpetologia, Instituto Butantan, Avenida Vital Brazil 1500, São Paulo 05503-900, SP, Brazil.
authors.forename: Wilson
authors.initials: W
authors.lastname: Fernandes
authors.affiliation: Laboratorio de Herpetologia, Instituto Butantan, Avenida Vital Brazil 1500, São Paulo 05503-900, SP, Brazil.
authors.forename: Anita Mitico
authors.initials: AM
authors.lastname: Tanaka-Azevedo
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
authors.forename: Julián
authors.initials: J
authors.lastname: Fernández
authors.affiliation: Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Laboratório de Hemostase e Venenos, Universidade Federal do Rio de Janeiro (UFRJ), Brazil; Instituto Nacional de Biologia Estrutural e Bioimagem, Rede Proteomica do Rio de Janeiro, Universidade Federal do Rio de Janeiro (UFRJ), Brazil. Electronic address: lzingali@bioqmed.ufrj.br.
authors.forename: Russolina B
authors.initials: RB
authors.lastname: Zingali
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
authors.forename: José María
authors.initials: JM
authors.lastname: Gutiérrez
authors.affiliation: Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Laboratório de Hemostase e Venenos, Universidade Federal do Rio de Janeiro (UFRJ), Brazil; Instituto Nacional de Biologia Estrutural e Bioimagem, Rede Proteomica do Rio de Janeiro, Universidade Federal do Rio de Janeiro (UFRJ), Brazil; Instituto Vital Brazil, Niterói, Rio de Janeiro, Brazil. Electronic address: netto@bioqmed.ufrj.br.
authors.forename: Carlos
authors.initials: C
authors.lastname: Corrêa-Netto
authors.affiliation: Laboratorio de Venómica Estructural y Funcional, Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain. Electronic address: jcalvete@ibv.csic.es.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
mesh_terms: D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D003435:Crotalid Venoms / Q000096:biosynthesis*; D005088:Exocrine Glands / Q000378:metabolism*; D020869:Gene Expression Profiling; D065947:Rainforest; D059467:Transcriptome / Q000502:physiology*
chemical_list: D003435:Crotalid Venoms
keywords: Antivenomics; Bothrops jararaca; Geographic venom variation; Snake venom proteomics; Venom gland transcriptomics