Standardization of in vitro nervous tissue culture for honeybee: A high specificity toxicological approach

Author(s)

Azevedo, Patricia ^{[1, 2]} ; Butolo, Nicole Pavan ^[1] ; de Alencar, Luciano Delmondes ^[2] ; Soares-Lima, Hellen Maria ^[1] ; Sales, Victor Ribeiro ^{[3, 4]} ; Malaspina, Osmar ^[1] ; Ferreira Nocelli, Roberta Cornelio ^{[3, 4]}

Affiliation

^[1] Univ Estadual Paulista Julio de Mesquita Filho UN, Dept Biol, CEIS, Programa Posgrad Biol Celular & Mol, Campus Rio Claro, Rio Claro, SP - Brazil

^[2] Univ Estadual Campinas UNICAMP, Inst Biol, Grp Genet & Genom Conservacao, Programa Posgrad Genet & Biol Mol, Campinas, SP - Brazil

^[3] Univ Fed Sao Carlos, Ctr Ciencias Agr, Dept Ciencias Nat Matemat & Educ, Grp Abelhas, Campus Araras, Sao Carlos, SP - Brazil

^[4] Univ Fed Sao Carlos, Ctr Ciencias Agr, Dept Ciencias Nat Matemat & Educ, Serv Ambientais, Campus Araras, Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY; v. 189, FEB 2020.
• Web of Science Citations: 0

Abstract

Bees are important pollinators that help to maintain the biodiversity of wild and cultivated plants. However, the increased and inappropriate use of agrochemicals has caused an imbalance in the populations of these insects visiting flowers for pollen and nectar collection. Therefore, new research methods for understanding the mechanisms of action of pesticides and their impacts on the brains of bees, such as neurotoxicity and cellular changes, in response to different active characteristics and dosages of insecticides are necessary. Thus, with the aim of developing tests with greater specificity at the level of cells or tissues, this study sought to standardize a method for the in vitro culture of the nervous tissue of Apis mellifera. For this purpose, the brains of six foragers bees were transferred to three different insect cell culture media and it supplementation with 10% foetal bovine serum (FBS): Grace, Schneider, Leibovitz, Grace + FBS, Schneider + FBS and Leibovitz + FBS media for each collection time. Nervous tissue was collected after 1, 6, 12 and 24 h of incubation in a humidified CO2 incubator at 32 degrees C, and histological sections of the organs were analysed. The results showed that Leibovitz medium and Leibovitz medium + serum are potential culture media for the cultivation of nervous tissue, since they resulted in less tissue spacing and tissue disarrangement. Therefore, additional supplements are necessary to obtain an ideal medium for the cultivation of A. mellifera nervous tissue. (AU)

• FAPESP's process: 14/12488-0 - Risk assessment of pesticides on bees and its impact on the pollination process
• Grantee: Roberta Cornélio Ferreira Nocelli
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/09555-5 - Synergistic effects between Nosema sp. and insecticide imidacloprid on honeybees africanized Apis mellifera
• Grantee: Hellen Maria Soares Lima
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/21097-3 - Bee-agriculture interactions: perspectives to sustainable use
• Grantee: Osmar Malaspina
• Support Opportunities: BIOTA-FAPESP Program - Thematic Grants