INFLUENCE OF THE PRESENCE OF CROTAMINE IN THE COMPOSITION OF THE CROTALUS DURISSUS TERRIFICUS VENOM FACE TO CROTALIC ANTIVENOM, A HISTOLOGICAL APPROACH

Authors

  • Fâni Ribeiro Silva Author
  • Nathalia Margarida Cantuária Author
  • Jonas Eligio Garcia de Azevedo Author
  • Gabriel Ferreira dos Santos Author
  • Jocimar de Souza Author
  • Leonardo da Silva Rodrigues Calderon Author
  • Isabella Ravazoli Author
  • José Carlos Cogo Author
  • Rafael Stuani Floriano Author
  • Stephen Hyslop Author
  • Yoko Oshima-Franco Author

DOI:

https://doi.org/10.56238/arev8n3-041

Keywords:

Antivenom, Crotalus durissus terrificus, Histology, Phrenic Nerve-Diaphragm Preparation, Snake Venom

Abstract

The main fractions isolated from Crotalus durissus terrificus venom include crotamine, gyrotoxin, crotoxin, convulxin and phospholipase A2. The antivenom is produced by a pool from Crotalus durissus ssp venoms, which do not always express crotamine, which may reflect on the treatment. The objective was to evaluate, by light microscopy, the effectiveness of antivenom in cellular injuries induced by C. d. terrificus venom with (Vcrot+) and without (Vcrot-) crotamine. The study design was in vitro using routine histological procedures on donated neuromuscular preparations under Ethics Committee 201/2021, exposed to 1) control; 2) Vcrot+; 3) Vcrot-; and treated with antivenom in 4) preincubation and 5) post-venom models. The results were myotoxicity index (MI) in control preparations of 42.2 % ± 2.6 cells; Vcrot+ 95.2 % ± 1.8; Vcrot- 89.6 % ± 2.3. In the preincubation model, the antivenom significantly decreased the lesions to 74.1 % ± 3.8 of Vcrot+ and 71.9 % ± 4.5 of Vcrot-. In the post-venom model, the results were Vcrot+ ® 10’, 98.0 % ± 0.1; ® 30’, 84.8 % ± 0.1; and ® 60’ 85.6 % ± 0.1, while for Vcrot- were 84.7 % ± 0.1; 83.6 % ± 0.01 and 91.9 % ± 0.09, respectively. In conclusion, the antivenom neutralizes a major amount of constituents in the venom in the preincubation model, while in the post-venom model, it was not able against the myotoxic effects of both venoms, but allowed to show the sequence of pathological events in an ex vivo model. 

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References

Aird, S. D., Kaiser, I. I., Lewis, R. V., & Kruggel, W. G. (1985). Rattlesnake presynaptic neurotoxins: Primary structure and evolutionary origin of the acidic subunit. Biochemistry, 24, 7054–7058. DOI: https://doi.org/10.1021/bi00346a005

Aird, S. D., Yates, J. R., Martino, P. A., Shabanowitz, J., Hunt, D. F., & Kaiser, I. I. (1990). The amino acid sequence of the acidic subunit B-chain of crotoxin. Biochimica et Biophysica Acta, 1040, 217–224. DOI: https://doi.org/10.1016/0167-4838(90)90079-U

Alexander, G., Grothusen, J., Zepeda, H., & Schwartzman, R. J. (1988). Gyroxin, a toxin from the venom of Crotalus durissus terrificus, is a thrombin-like enzyme. Toxicon, 26, 953–960. DOI: https://doi.org/10.1016/0041-0101(88)90260-7

Azevedo-Marques, M. M., Hering, S. E., & Cupo, P. (1987). Evidence that Crotalus durissus terrificus (South American rattlesnake) envenomation in humans causes myolysis rather than hemolysis. Toxicon, 25, 1163–1168. DOI: https://doi.org/10.1016/0041-0101(87)90134-6

Azevedo-Marques, M. M., Hering, S. E., & Cupo, P. (2003). Crotalic accident. In J. L. C. Cardoso, O. S. F. França, F. H. Wen, C. M. S. Málaque, & V. Haddad Jr. (Eds.), Venomous animals in Brazil: Biology, clinic and therapeutics of accidents (pp. 91–98). Sarvier.

Barrabin, H., Martiarena, J. L., Vidal, J. C., & Barrio, A. (1978). Isolation and characterization of gyroxin from Crotalus durissus terrificus venom. In P. Rosemberg (Ed.), Toxins: Animal, plant, microbial (pp. 113–133). Pergamon. DOI: https://doi.org/10.1016/B978-0-08-022640-8.50017-2

Barros, L. C., Soares, A. M., Costa, F. L., Rodrigues, V. M., Fuly, A. L., Giglio, J. R., Gallacci, M., Thomazini-Santos, I. A., Barraviera, S. R. C. S., Barraviera, B., & Ferreira Junior, R. S. (2011). Biochemical and biological evaluation of gyroxin isolated from Crotalus durissus terrificus venom. Journal of Venomous Animals and Toxins including Tropical DOI: https://doi.org/10.1590/S1678-91992011000100004

Diseases, 17, 23–33.

Batista da Cunha, D., Pupo Silvestrini, A. V., Gomes da Silva, A. C., Maria de Paula Estevam, D., Pollettini, F. L., de Oliveira Navarro, J., Alves, A. A., Remédio Zeni Beretta, A. L., Annichino Bizzacchi, J. M., Pereira, L. C., & Mazzi, M. V. (2018). Mechanistic insights into functional characteristics of native crotamine. Toxicon, 146, 1–12. DOI: https://doi.org/10.1016/j.toxicon.2018.03.007

Brazil, Ministry of Health. (2001). Manual for diagnosing and treating accidents caused by venomous animals - National Health Foundation (2nd ed.). https://www.gov.br/saude/pt-br/assuntos/saude-de-a-a-z/a/animais-peconhentos/aguas-vivas-e-caravelas/publicacoes/manual-de-diagnostico-e-tratamento-de-acidentes-por-animais-peconhentos.pdf/view

Brazil, Ministry of Health. (2018). Brazil: Epidemiological bulletin. Snake accidents in Brazil; Health Surveillance Secretariat (Vol. 51, No. 09). https://www.gov.br/saude/pt-br/assuntos/saude-de-a-a-z/a/animais-peconhentos/acidentes-por-abelhas/arquivos/boletim-epidemiologico-09-vol-51mar-2020-acidentes-ofidicos-no-brazil-2018.pdf/view

Brazil, Ministry of Health. (2022). Brazil: Notifiable Diseases Information System - Sinan Net.: Accidents caused by venomous animals - notifications registered in the information system for notifiable diseases. http://tabnet.datasus.gov.br/cgi/tabcgi.exe?sinannet/cnv/animaisbr.def

Brazil, O. V., & Excell, B. J. (1971). Action of crotoxin and crotactin from the venom of Crotalus durissus terrificus (South American rattlesnake) on the frog neuromuscular junction. The Journal of Physiology, 212, 34–35.

Brazil, O. V., & Fontana, M. D. (1993). Toxins as tools in the study of sodium channel distribution in the muscle fibre membrane. Toxicon, 31, 1085–1098. DOI: https://doi.org/10.1016/0041-0101(93)90124-2

Camargo, T. M., Nazato, V. S., Silva, M. G., Cogo, J. C., Groppo, F. C., & Oshima-Franco, Y. (2010). Bothrops jararacussu venom-induced neuromuscular blockade inhibited by Casearia gossypiosperma Briquet hydroalcoholic extract. Journal of Venomous Animals and Toxins including Tropical Diseases, 16, 432–441. DOI: https://doi.org/10.1590/S1678-91992010000300009

Cameron, D. L., & Tu, A. T. (1978). Chemical and functional homology of myotoxin a from prairie rattlesnake venom and crotamine from South American rattlesnake venom. Biochimica et Biophysica Acta, 532, 147–154. DOI: https://doi.org/10.1016/0005-2795(78)90457-9

Carvalho, L. H., Teixeira, L. F., Zaqueo, K. D., Bastos, J. F., Nery, N. M., Setúbal, S. S., Pontes, A. S., Butzke, D., Cavalcante, W., Gallacci, M., Fernandes, C. F. C., Stabeli, R. G., Soares, A. M., & Zuliani, J. P. (2019). Local and systemic effects caused by Crotalus durissus terrificus, Crotalus durissus collilineatus, and Crotalus durissus cascavella snake venoms in Swiss mice. Revista da Sociedade Brasileira de Medicina Tropical, 52, Article e20180526. DOI: https://doi.org/10.1590/0037-8682-0526-2018

Cheymol, J., Gonçalves, J. M., Bourillet, F., & Roch-Arveiller, M. (1971). Action neuromusculaire comparée de la crotamine et du venin de Crotalus durissus terrificus var. crotaminicus. 1. Sur préparations neuromusculaires in situ [A comparison of the neuromuscular action of crotamine and the venom of Crotalus durissus terrificus var. crotaminicus. 1. Neuromuscular preparations in situ]. Toxicon, 9, 279–286. DOI: https://doi.org/10.1016/0041-0101(71)90081-X

Cupo, P., Azevedo-Marques, M. M., & Hering, S. E. (1990). Acute myocardial infarction-like enzyme profile in human victims of Crotalus durissus terrificus envenoming. Transactions of the Royal Society of Tropical Medicine and Hygiene, 84, 447–451. DOI: https://doi.org/10.1016/0035-9203(90)90358-L

Da Silva-Júnior, L. N., Abreu, L. S., Rodrigues, C. F. B., Galizio, N. D. C., Aguiar, W. D. S., Serino-Silva, C., Dos Santos, V. S., Costa, I. A., Oliveira, L. V. F., Sant’Anna, S. S., Grego, K. F., Tanaka-Azevedo, A. M., Rodrigues, L. N. D. S., & de Morais-Zani, K. (2020). Geographic variation of individual venom profile of Crotalus durissus snakes. Journal of Venomous Animals and Toxins including Tropical Diseases, 26, Article e20200016. DOI: https://doi.org/10.1590/1678-9199-jvatitd-2020-0016

De Oliveira, S. A., Magalhães, M. R., Salazar, V. C., Valadares, M. C., & da Cunha, L. C. (2015). Identification of crotamine in the venom of Crotalus durissus collilineatus by three different methods. Toxicon, 95, 46–51. DOI: https://doi.org/10.1016/j.toxicon.2014.12.015

De Souza, J., Oliveira, I. C. F., Yoshida, E. H., Cantuaria, N. M., Cogo, J. C., Torres-Bonilla, K. A., Hyslop, S., Silva Junior, N. J., Floriano, R. S., Gutiérrez, J. M., & Oshima-Franco, Y. (2022). Effect of the phospholipase A2 inhibitor Varespladib, and its synergism with crotalic antivenom, on the neuromuscular blockade induced by Crotalus durissus terrificus venom (with and without crotamine) in mouse neuromuscular preparations. Toxicon, 214, 54–61. DOI: https://doi.org/10.1016/j.toxicon.2022.05.001

Ferraz, M. C., Yoshida, E. H., Tavares, R. V., Cogo, J. C., Cintra, A. C., Dal Belo, C. A., Franco, L. M., dos Santos, M. G., Resende, F. A., Varanda, E. A., Hyslop, S., Puebla, P., San Feliciano, A., & Oshima-Franco, Y. (2014). An isoflavone from Dipteryx alata Vogel is active against the in vitro neuromuscular paralysis of Bothrops jararacussu snake venom and bothropstoxin I, and prevents venom-induced myonecrosis. Molecules, 19, 5790–5805. DOI: https://doi.org/10.3390/molecules19055790

Franco, Y. O., & Belo, C. D. (2017). Recognizing antiophidian plants using the neuromuscular junction apparatus. International Journal of Complementary & Alternative Medicine, 5, 11–12. DOI: https://doi.org/10.15406/ijcam.2017.05.00165

Fusco, L. S., Rodríguez, J. P., Teibler, P., Maruñak, S., Acosta, O., & Leiva, L. (2015). New immunization protocol to produce crotalic antivenom combining Crotalus durissus terrificus venom and its PLA2. Biologicals, 43, 62–70. DOI: https://doi.org/10.1016/j.biologicals.2014.09.001

Ginn, F. L., Shelburne, J. D., & Trump, B. F. (1968). Disorders of cell volume regulation. I. Effects of inhibition of plasma membrane adenosine triphosphatase with ouabain. The American Journal of Pathology, 53, 1041–1071.

Gonçalves, J. M., & Vieira, L. G. (1950). Studies on Brazilian snake venoms. I – Electrophoretic analysis. Anais da Academia Brasileira de Ciências, 22, 141–150.

Gonçalves, J. M. (1956). Studies on Brazilian snake venoms. II - Crotalus terrificus crotaminicus, subespécie biológica. Anais da Academia Brasileira de Ciências, 28, 365–367.

Gutiérrez, J. M., & Cerdas, L. (1984). Mechanism of action of myotoxins isolated from poisons from snakes [Mechanism of action of myotoxins isolated from snake venoms]. Revista de Biología Tropical, 32, 213–222.

Gutiérrez, J. M., León, G., Rojas, G., Lomonte, B., Rucavado, A., & Chaves, F. (1998). Neutralization of local tissue damage induced by Bothrops asper (terciopelo) snake venom. Toxicon, 36, 1529–1538. DOI: https://doi.org/10.1016/S0041-0101(98)00145-7

Gutiérrez, J. M., & Lomonte, B. (1995). Phospholipase A2 myotoxins from Bothrops snake venoms. Toxicon, 33, 1405–1424. DOI: https://doi.org/10.1016/0041-0101(95)00085-Z

Hampe, O. G., Vozári-Hampe, M. M., & Gonçalves, J. M. (1978). Crotamine conformation: effect of pH and temperature. Toxicon, 16, 453–460. DOI: https://doi.org/10.1016/0041-0101(78)90142-3

Hörtnagel, H., & Hanin, I. (1992). Toxins affecting the cholinergic system. In H. Herken & F. Hucho (Eds.), Selective neurotoxicity (pp. 293–332). Berlin. DOI: https://doi.org/10.1007/978-3-642-85117-9_9

Iglesias, C. V., Aparicio, R., Rodrigues-Simioni, L., Camargo, E. A., Antunes, E., Marangoni, S., de Oliveira Toyama, D., Beriam, L. O., Monteiro, H. S., & Toyama, M. H. (2005). Effects of morin on snake venom phospholipase A2 (PLA2). Toxicon, 46, 751–758. DOI: https://doi.org/10.1016/j.toxicon.2005.07.017

Kouyoumdjian, J. A., Harris, J. B., & Johnson, M. A. (1986). Muscle necrosis caused by the sub-units of crotoxin. Toxicon, 24, 575–583. DOI: https://doi.org/10.1016/0041-0101(86)90178-9

Kruse, H., Van Wimersma Greidanus, T. B., & De Wied, D. (1977). Barrel rotation induced by vasopressin and related peptides in rats. Pharmacology Biochemistry and Behavior, 7, 311–313. DOI: https://doi.org/10.1016/0091-3057(77)90226-X

Lago, L. A., Ferreira, P. M., Facury Filho, E. J., Melo, M. M., & Alzamoura Filho, F. (2000). Clinical picture of experimental crotal poisoning in cattle (Crotalus durissus terrificus - crotamine positive). Brazilian Journal of Veterinary Research and Animal Science, 37, 312–315. DOI: https://doi.org/10.1590/S1413-95962000000400010

Laure, C. J. (1975). Die primärstruktur des crotamins [The primary structure of crotamine (author's transl)]. Hoppe-Seyler's Zeitschrift für Physiologische Chemie, 356, 213–215. DOI: https://doi.org/10.1515/bchm2.1975.356.1.213

Lima, S. C., Porta, L. C., Lima, Á. D. C., Campeiro, J. D., Meurer, Y., Teixeira, N. B., Duarte, T., Oliveira, E. B., Picolo, G., Godinho, R. O., Silva, R. H., & Hayashi, M. A. F. (2018). Pharmacological characterization of crotamine effects on mice hind limb paralysis employing both ex vivo and in vivo assays: Insights into the involvement of voltage-gated ion channels in the crotamine action on skeletal muscles. PLoS Neglected Tropical Diseases, 12, Article e0006700. DOI: https://doi.org/10.1371/journal.pntd.0006700

Lomonte, B., Angulo, Y., & Calderón, L. (2003). An overview of lysine-49 phospholipase A2 myotoxins from crotalid snake venoms and their structural determinants of myotoxic action. Toxicon, 42, 885–901. DOI: https://doi.org/10.1016/j.toxicon.2003.11.008

Lourenço, A. Jr., Zorzella Creste, C. F., de Barros, L. C., Delazari dos Santos, L., Pimenta, D. C., Barraviera, B., & Ferreira, R. S. Jr. (2013). Individual venom profiling of Crotalus durissus terrificus specimens from a geographically limited region: crotamine assessment and captivity evaluation on the biological activities. Toxicon, 69, 75–81. DOI: https://doi.org/10.1016/j.toxicon.2013.01.006

Melgarejo, A. R. (2009). Venomous snakes of Brazil. In J. L. C. Cardoso, S. F. O. França, F. H. Wen, C. M. S. Málaque, & V. Haddad Junior (Eds.), Venomous animals in Brazil: Biology, clinic and therapeutics of poisoning (pp. 61–62).

Mebs, D., & Ownby, C. L. (1990). Myotoxic components of snake venoms: Their biochemical and biological activities. Pharmacology & Therapeutics, 48, 223–236. DOI: https://doi.org/10.1016/0163-7258(90)90081-C

Montecucco, C., Gutiérrez, J. M., & Lomonte, B. (2008). Cellular pathology induced by snake venom phospholipase A2 myotoxins and neurotoxins: Common aspects of their mechanisms of action. Cellular and Molecular Life Sciences, 65, 2897–2912. DOI: https://doi.org/10.1007/s00018-008-8113-3

Nepal, M. R., & Jeong, T. C. (2020). Alternative methods for testing botulinum toxin: Current status and future perspectives. Biomolecules & Therapeutics, 28, 302–310. DOI: https://doi.org/10.4062/biomolther.2019.200

Oguiura, N., Boni-Mitake, M., & Rádis-Baptista, G. (2005). New view on crotamine, a small basic polypeptide myotoxin from South American rattlesnake venom. Toxicon, 46, 363–370. DOI: https://doi.org/10.1016/j.toxicon.2005.06.009

Oliveira, K. C. (2014). Biochemical and pharmacological characterization of crotamine irradiated by gamma rays in co-60 [Tese de doutorado, Instituto de Pesquisas Energéticas e Nucleares - Universidade de São Paulo].

Ownby, C. L., Aird, S. D., & Kaiser, I. I. (1988). Physiological and immunological properties of small myotoxins from the venom of the midget faded rattlesnake (Crotalus viridis concolor). Toxicon, 26, 319–323. DOI: https://doi.org/10.1016/0041-0101(88)90223-1

Peigneur, S., Orts, D. J., Prieto da Silva, A. R., Oguiura, N., Boni-Mitake, M., de Oliveira, E. B., Zaharenko, A. J., de Freitas, J. C., & Tytgat, J. (2012). Crotamine pharmacology revisited: Novel insights based on the inhibition of KV channels. Molecular Pharmacology, 82, 90–96. DOI: https://doi.org/10.1124/mol.112.078188

Pereira, A. (2011). Selective toxicity of crotamine from Crotalus durissus terrificus venom on tumor-inducing cells [Tese de doutorado, Universidade de São Paulo].

Pinho, F. M. O., & Pereira, I. D. (2001). Ophidism. Revista da Associação Médica Brasileira, 47, 24–29. DOI: https://doi.org/10.1590/S0104-42302001000100026

Prado-Franceschi, J., & Brazil, O. V. (1981). Convulxin, a new toxin from the venom of the South American rattlesnake Crotalus durissus terrificus. Toxicon, 19, 875–887. DOI: https://doi.org/10.1016/0041-0101(81)90085-4

PubChem. (n.d.). Crotamin (CID 85470858). National Center for Biotechnology Information. https://pubchem.ncbi.nlm.nih.gov/compound/Crotamin#section=2D-Structure

Rodrigues, M., Santos, A., de la Torre, B. G., Rádis-Baptista, G., Andreu, D., & Santos, N. C. (2012). Molecular characterization of the interaction of crotamine-derived nucleolar targeting peptides with lipid membranes. Biochimica et Biophysica Acta, 1818, 2707–2717. DOI: https://doi.org/10.1016/j.bbamem.2012.06.014

Rosenfeld, G. (1971). Symptomatology, pathology, treatment of snake bites in South America. In W. Bucherl & E. E. Buckley (Eds.), Venomous animals and their venoms (pp. 345–384). New York. DOI: https://doi.org/10.1016/B978-0-12-138902-4.50021-0

Sampaio, S. C., Hyslop, S., Fontes, M. R., Prado-Franceschi, J., Zambelli, V. O., Magro, A. J., Brigatte, P., Gutierrez, V. P., & Cury, Y. (2010). Crotoxin: Novel activities for a classic beta-neurotoxin. Toxicon, 55, 1045–1060. DOI: https://doi.org/10.1016/j.toxicon.2010.01.011

Silvestrini, A. V. P., de Macedo, L. H., de Andrade, T. A. M., Mendes, M. F., Pigoso, A. A., & Mazzi, M. V. (2019). Intradermal application of crotamine induces inflammatory and immunological changes in vivo. Toxins, 11, Article 39. DOI: https://doi.org/10.3390/toxins11010039

Siqueira, A. M., Martins, N. F., De Lima, M. E., Diniz, C. R., Cartier, A., Brown, D., & Maigret, B. (2002). A proposed 3D structure for crotamine based on homology building, molecular simulations and circular dichroism. Journal of Molecular Graphics and Modelling, 20, 389–398. DOI: https://doi.org/10.1016/S1093-3263(01)00139-5

Slotta, K. H., & Fraenkel-Conrat, H. (1938). Two active proteins from rattlesnake venom. Nature, 142, 213.

Thakshila, P., Hodgson, W. C., Isbister, G. K., & Silva, A. (2022). In vitro neutralization of the myotoxicity of Australian mulga snake (Pseudechis australis) and Sri Lankan Russell's Viper (Daboia russelii) venoms by Australian and Indian polyvalent antivenoms. Toxins, 14, Article 302. DOI: https://doi.org/10.3390/toxins14050302

Tasima, L. J., Silva, C. S., Hatakeyama, D. M., Nishiduka, E. S., Tashima, A. K., Sant’Anna, S. S., Grego, K. F., Zani, K. M., & Azevedo, A. M. T. (2020). Crotamine in Crotalus durissus: Distribution according to subspecies and geographic origin, in captivity or nature. Journal of Venomous Animals and Toxins including Tropical Diseases, 6, Article e20190053. DOI: https://doi.org/10.1590/1678-9199-jvatitd-2019-0053

Teixeira-Araújo, R., Castanheira, P., Brazil-Más, L., Pontes, F., Leitão de Araújo, M., Machado Alves, M. L., Zingali, R. B., & Correa-Netto, C. (2017). Antivenomics as a tool to improve the neutralizing capacity of the crotalic antivenom: A study with crotamine. Journal of Venomous Animals and Toxins including Tropical Diseases, 12, 23–28. DOI: https://doi.org/10.1186/s40409-017-0118-7

Toyama, M. H., Marangoni, S., Novello, J. C., Leite, G. B., Prado-Franceschi, J., da Cruz-Höfling, M. A., & Rodrigues-Simioni, L. (2003). Biophysical, histopathological and pharmacological characterization of crotamine isoforms F22 and F32. Toxicon, 41, 493–500. DOI: https://doi.org/10.1016/S0041-0101(02)00390-2

Vonk, F. J., Jackson, K., Doley, R., Madaras, F., Mirtschin, P. J., & Vidal, N. (2011). Snake venom: From fieldwork to the clinic: Recent insights into snake biology, together with new technology allowing high-throughput screening of venom, bring new hope for drug discovery. BioEssays, 33, 269–279. DOI: https://doi.org/10.1002/bies.201000117

World Health Organization. (2023). Snakebite envenoming. https://www.who.int/news-room/fact-sheets/detail/snakebite-envenoming

World Health Organization. (2018, August 19). WHO issues new recommendation on antivenom for snakebites. https://www.who.int/news/item/19-08-2018-who-issues-new-recommendation-on-antivenom-for-snakebites

Yamane, E. S., Bizerra, F. C., Oliveira, E. B., Moreira, J. T., Rajabi, M., Nunes, G. L., de Souza, A. O., da Silva, I. D., Yamane, T., Karpel, R. L., Silva, P. I. Jr., & Hayashi, M. A. (2013). Unraveling the antifungal activity of a South American rattlesnake toxin crotamine. Biochimie, 95, 231–240. DOI: https://doi.org/10.1016/j.biochi.2012.09.019

Zanetti, G., Negro, S., Pirazzini, M., & Caccin, P. (2018). Mouse phrenic nerve hemidiaphragm assay (MPN). Bio-protocol, 8, Article e2759. DOI: https://doi.org/10.21769/BioProtoc.2759

Slotta, K. H., & Fraenkel-Conrat, H. (1938). Chemical studies on snake venoms. Purification and crystallization of rattlesnake venom. Nature, 142, 213. DOI: https://doi.org/10.1038/142213a0

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SILVA, Fâni Ribeiro et al. INFLUENCE OF THE PRESENCE OF CROTAMINE IN THE COMPOSITION OF THE CROTALUS DURISSUS TERRIFICUS VENOM FACE TO CROTALIC ANTIVENOM, A HISTOLOGICAL APPROACH. ARACÊ , [S. l.], v. 8, n. 3, p. e12462, 2026. DOI: 10.56238/arev8n3-041. Disponível em: https://periodicos.newsciencepubl.com/arace/article/view/12462. Acesso em: 14 mar. 2026.