Optimization of in vitro culture of honeybee nervous tissue for pesticide risk assessment

Author(s)

Azevedo, Patricia ; Butolo, Nicole Pavan ; de Alencar, Luciano Delmondes ; Lima, Hellen Maria Soares ; Sales, Victor Ribeiro ; Malaspina, Osmar ; Nocelli, Roberta Cornelio Ferreira

• Document type: Journal article
• Source: TOXICOLOGY IN VITRO; v. 84, p. 14-pg., 2022-10-01.

Abstract

The most used pesticides have neurotoxic action on the neurotransmitter system of target and non-targeted insects, such as honeybees. However, honeybees have foremost importance worldwide, which has encouraged the development of tools to evaluate the action of specific pesticide molecules on their nervous system, providing accurate data on damage to their brain. In this sense, our study aimed to optimize in vitro honeybee nervous tissue culture to assess pesticide risks. To this end, six forager honeybee brains were dissected and transferred to different combinations of Leibovitz-15 (L-15) culture medium supplemented with Fetal Bovine Serum (FBS), Hank's Balanced Salt Solution (HBSS), and Insect Medium Supplement (IMS). Nervous tissues were collected after different incubation times (1, 6, 12, and 24 h) for morphology and Kenyon cell analyses. Our results showed that L-15 medium supplemented with HBSS and with HBSS plus FBS were the best media for culturing honey nervous tissue for 24 h, as they resulted in less tissue spacing and cell disarrangement. Therefore, they may be assessed in future ecotoxicological tests. (AU)

• FAPESP's process: 17/21097-3 - Bee-agriculture interactions: perspectives to sustainable use
• Grantee: Osmar Malaspina
• Support Opportunities: BIOTA-FAPESP Program - Thematic Grants
• FAPESP's process: 18/24245-6 - Genomic, transcriptomic and metagenomic in populations of native stingless bees aiming the search for detoxification genes
• Grantee: Patricia Azevedo
• Support Opportunities: Scholarships in Brazil - Doctorate