Impact of environmental concentrations of fipronil on DNA integrity and brain structure of Bombus atratus bumblebees.

Fipronil (FP) is an insecticide used in the treatment and control of pests, but it also adversely affects bees. Currently, there is no data on the genotoxic effects of FP in the brain of bumblebees. Thus, through the comet assay and routine morphological analysis, we analyzed the morphological effects and potential genotoxicity of environmentally relevant concentrations of FP on the brain of Bombus atratus. Bumblebees were exposed at concentrations of 2.5 µg/g and 3.5 µg/g for 96 hours. After the exposure, the brains were removed for morphological and morphometric analysis, and the comet assay procedure - used to detect DNA damage in individual cells using electrophoresis. Our data showed that both concentrations (2.5 µg/g and 3.5 µg/g) caused DNA damage in brain cells. These results corroborate the morphological data. We observed signs of synapse loss in the calyx structure, intercellular spaces between compact inner and non-compact inner cells, and cell swelling. This study provides unprecedented evidence of the effects of FP on DNA and cellular structures in the brain of B. atratus and reinforces the need to elucidate its toxic effects on other species to allow future risk assessments and conservation projects.

Genotoxic effect of two environmentally safe doses of cadmium on the hepato-nephrocytic system of Bombus atratus forager workers.

Eusocial bees are declining due to anthropogenic actions. Individuals can be exposed to contaminants like Cd, which have been found in pollen. Thus, we evaluated the potential genotoxicity of Cd to the hepato-nephrocitic system of Bombus atratus foraging workers exposed to environmentally safe doses of Cd (0.001 mg/kg and 0.003 mg/kg) during 72 h. To assess the genotoxicity of Cd, we conducted an alkaline comet assay. Doses of 0.001 mg/kg of Cd caused comets of levels 2 and 3, and 0.003 mg/kg of Cd induced comets of level 4, while controls exhibited regular nucleoids. Also, 0.003 mg/kg doses caused higher Damage Index, Damage Frequency, and Total Damage. Our results bring new evidence that 0.003 mg/kg Cd exerted genotoxicity to the HNS cells of B. atratus, probably affecting secondary metabolism and the detoxification capability of bees.

Effects of mercury at field estimated concentration in brain of Bombus atratus (Hymenoptera: Bombini).

Although mercury neurotoxic effects are well known in several species, it is poorly studied in bees. Mercury contamination is increasing in several regions of the Brazilian Amazon Rainforest due to illegal and indiscriminate gold mining. Therefore, this study aimed to evaluate the effects of mercury (Hg) in brain Kenyon cells of foraging workers of Bombus atratus exposed to an average concentration (110 ppb) found in pots of honey from native bees of South America and Australia. Twenty forager workers were collected in the field (23° 34' S 47° 31' W), divided into control (n = 10) and exposed (n = 10) groups, and individually kept in special boxes for 48 h. For the exposed group, we offered Hg solution (at 110 ppb) ad libitum, while for the control group we offered water, and for both sucrose syrup at 70%. After the exposure time, the bees were crio-anesthezied at 4 °C. Brains were dissected and processed for morphological, morphometric, and histochemical analyses. Morphological results showed that the Kenyon cells of the Hg-exposed group presented both cytoplasmic vacuolization and nuclear pyknosis, which indicate cell death. These findings were corroborated by the acridine orange staining. Hg exposure also induced significant nuclear chromatin compaction in Kenyon cells. The calyces and peduncles of the mushroom bodies showed disorganization and vacuolization. In summary, these changes may imply in a severe impairment of the cognitive abilities of the bees, which could lead them to the loss of many tasks, such as foraging or even nest founding by the queen.