Expression and function of the vitellogenin receptor in the hypopharyngeal glands of the honey bee Apis mellifera (Hymenoptera: Apidae) workers.

The vitellogenin receptor (VgR) is essential for the uptake and transport of the yolk precursor, vitellogenin (Vg). Vg is synthesized in the fat body, released in the hemolymph, and absorbed in the ovaries, via receptor-mediated endocytosis. Besides its important role in the reproductive pathway, Vg occurs in nonreproductive worker honey bee, suggesting its participation in other pathways. The objective was to verify if the VgR occurs in the hypopharyngeal glands of Apis mellifera workers and how Vg is internalized by these cells. VgR occurrence in the hypopharyngeal glands was evaluated by qPCR analyses of VgR and immunohistochemistry in workers with different tasks. The VgR gene is expressed in the hypopharyngeal glands of workers with higher transcript levels in nurse honey bees. VgR is more expressed in 11-day-old workers from queenright colonies, compared to orphan ones. Nurse workers with developed hypopharyngeal glands present higher VgR transcripts than those with poorly developed glands. The immunohistochemistry results showed the co-localization of Vg, VgR and clathrin (protein that plays a major role in the formation of coated vesicles in endocytosis) in the hypopharyngeal glands, suggesting receptor-mediated endocytosis. The results demonstrate that VgR performs the transport of Vg to the hypopharyngeal glands, supporting the Ovary Ground Plan Hypothesis and contributing to the understanding of the role of this gland in the social context of honey bees.

Acute oral exposure to imidacloprid induces apoptosis and autophagy in the midgut of honey bee Apis mellifera workers.

The honey bee Apis mellifera is an important pollinator that increases the yield and quality of crops. In recent years, honey bee populations have declined in some parts of the world, which has been associated with several causes, including pesticides used in agriculture. Neonicotinoids are neurotoxic insecticides widely used in the world with systemic action mode contaminating nectar and pollen that may be consumed by bees. This study evaluated the side effects of imidacloprid in the midgut of A. mellifera after acute oral exposure. Toxicity, histopathology, cytotoxicity, and expression of autophagy-related gene atg1 were evaluated in honey bee workers orally exposed to imidacloprid. The estimated imidacloprid LC50 was 1.44 mg L-1. The midgut epithelium of bees fed on imidacloprid LC50 has the occurrence of cytoplasm vacuoles, enlarged intercellular spaces, disorganization of the striated border, and nuclear pyknosis, with an organ injury index that increases with time exposure. The midgut digestive cells of treated bees have apical protrusions, damaged mitochondria, and autophagosomes that were characterized for content with organelle debris and high expression of atg1. These features indicate the occurrence of high cell death in the midgut of workers exposed to imidacloprid, which may affect the digestibility the physiology of the insect.

Cuticle melanization and the expression of immune-related genes in the honeybee Apis mellifera (Hymenoptera: Apidae) adult workers.

The global decline of bee populations has several factors, including pathogens, which need overcome the insect defenses such as the physical barriers, the body cuticle and peritrophic matrix (primary defenses), as well as the secondary defenses with antimicrobial peptides (AMPs) and the enzyme lysozyme. The regulation of immune defenses according to the infection risks raises questions about the immunity of social bees due to their exposition to different pathogens pressures during the adult lifespan and tasks performed. This study evaluated the primary (body cuticle melanization, peritrophic matrix and cpr14 expression) and secondary (AMPs and lysozyme expression) defenses of the honeybee Apis mellifera workers according to the age and tasks. The expression of malvolio was used to detect precocious forage tasks outside the colony. Forager workers have higher amount of cuticular melanization in the body cuticle than nurse, but not when the age effect is retired, indicating the gradual acquisition of this compound in the integument of adult bees. The relative value of chitin in the peritrophic matrix and cpr14 mRNA are similar in all bees evaluated, suggesting that these components of primary defenses do not change according to the task and age. Differential expression of genes for AMPs in workers performing different tasks, within the same age group, indicates that the behavior stimulates expression of genes related to secondary immune defense. The expression of malvolio gene, accelerating the change in workers behavior, and those related to immune defense suggest the investment in secondary defense mechanisms when the primary defense of the body cuticle is not yet completed.

Differential expression of aquaporin genes during ovary activation in the honeybee Apis mellifera (Hymenoptera: Apidae) queens.

Aquaporins are membrane proteins responsible for the rapid transport of water and solutes through the plasma membrane in almost all organisms including microorganisms, animals and plants. The knowledge about aquaporins in insects has advanced in the last years, especially in organs such as the gut and Malpighian tubules, but their roles in ovaries are poorly understood. This study reports the occurrence and the expression of gene profiles of predicted aquaporins in the ovaries of virgin and mated queens of the honeybee Apis mellifera. The expression of six aquaporin genes (Am-DRIP, Am-PRIP, Am-Eglp1, Am-Eglp2, Am-Eglp3 and Am-BIB) was investigated in the ovaries of queens of different ages and physiological status. The Am-Eglp3 has the higher level of transcript in newly emerged virgin queens. The Am-PRIP and Am-Eglp2 have a higher level of transcripts in 4 days-old virgin queens, aged expected to mate. In mated queens, only the Am-Eglp2 show high level of transcripts. Aquaporin was immunolocalized in the follicular cells of both nurse and oocytic chambers of the previtellogenic and vitellogenic follicles. This is the first report for the presence of aquaporins in the ovaries of bees.

Aprendizagem associativa em fêmeas e machos de Anastrepha obliqua (Diptera, Tephritidae) com diferentes idades e estados nutricionais / Associative learning in anastrepha obliqua (diptera, tephritidae) wild females and males of different ages and nutritional states

Adult Anastrepha obliqua fruit-flies need protein sources to promote sexual maturation and egg production. Learning can be a mechanism to improve the foraging behavior to acquire this nutrient. Learning could be defined as the acquisition of a new representation of neuronal information, and it can only be indirectly observed through its potential effect on behavior. In the present study, the occurrence or not of associative learning in A. obliqua females and males of different ages and nutritional states concerning sucrose (carbohydrate) and brewer´s yeast (protein source) was observed. Associative learning for sucrose was demonstrated in young and old females and males only when deprived of food for 2 days after the experience period. As concerns yeast, the associative learning was demonstrated in deprived young and old females only.

Adultos de moscas-das-frutas, Anastrepha obliqua, necessitam de fontes proteicas para promover a maturação sexual e produção de ovos. A aprendizagem pode ser um mecanismo para melhorar o comportamento de forrageamento na aquisição deste nutriente. Dentre as diferentes definições de aprendizagem uma delas a define como a aquisição de uma nova representação de informação neuronal e pode ser indiretamente observada através de seu efeito potencial no comportamento. No presente estudo, a ocorrência ou não de aprendizagem em fêmeas e machos de A. obliqua de diferentes idades e estados nutricionais foram observadas em relação à sacarose (carboidrato) e lêvedo de cerveja (fonte de proteína). A aprendizagem associativa para sacarose foi demonstrada em fêmeas e machos jovens e velhos somente quando privados de alimento por dois dias depois do período de experiência. Em relação ao lêvedo, a aprendizagem associativa foi demonstrada em fêmeas jovens e velhas, mas não nos machos.