Efeitos tardios da co exposição larval a doses realísticas do inseticida clotianidina e do fungicida piraclostrobina : avaliação da longevidade das operárias adultas de apis mellifera africanizada

Nos últimos anos, as abelhas vêm sofrendo um declínio populacional que afeta os serviços ecossistêmicos de polinização, dentre os fatores desse declínio estão os agrotóxicos. Com o objetivo de avaliar o efeito da exposição a uma mistura do inseticida clotianidina e do fungicida piraclostrobina no desenvolvimento de Apis mellifera africanizada, larvas de primeiro instar obtidas de três colônias foram transferidas para cúpulas com dieta artificial padronizada. Realizou-se exposição repetida por via oral nas larvas, do terceiro ao sexto dia do bioensaio. O consumo total do inseticida foi de 0,2364 ng/larva e do fungicida de 23,63 ng/larva. Avaliou-se a longevidade das abelhas emergidas e realizou a caracterização química dos compostos por infravermelho. Os agrotóxicos não apresentaram efeitos deletérios nas larvas e pupas. Entretanto, as abelhas expostas ao inseticida isolado e aos dois agrotóxicos combinados apresentaram redução na longevidade dos adultos emergidos (p<0,01). O decréscimo no tempo de sobrevivência foi maior no grupo exposto apenas ao inseticida, sendo que a presença do fungicida no grupo co-exposto atenuou este decréscimo na longevidade, possivelmente por uma interação dos compostos químicos dos agrotóxicos, evidenciada pela mudança no padrão do espectrograma. O fungicida isolado não alterou a longevidade das abelhas (p>0,1). A co-exposição pode ter diminuído a toxicidade do inseticida por meio de interações químicas e/ou facilitado o processo de desintoxicação destes xenobióticos no organismo, reduzindo a mortalidade. Do ponto de vista da dinâmica populacional da colônia, estes dados são preocupantes, visto que as concentrações dos agrotóxicos foram encontradas nos recursos alimentares das abelhasFil: Menezes Tadei, Rafaela. Universidade Federal de Sao Carlos (Brasil)Fil: Aparecido, J.. Universidade Federal de Sao Carlos (Brasil)Fil: Silva Zacarin, Elaine C. M.. Universidade Federal de Sao Carlos (Brasil

Fipronil effect on the frequency of anomalous brood in honeybee reared in vitro

Larvae of honeybee workers were exposed to the insecticide fipronil during the feeding phase. To evaluate the effect of fipronil in the post-embryonic development of africanized Apis mellifera, bioassays of toxicity were done. The bioassays were performed by acute exposure applying 1μL of distilled water for control (I) and for experiments: 0.5 ng a.i./μL of fipronil; 5 ng a.i./μL of fipronil and 20 ng a.i./ μL of fipronil. Triplicates were performed for all treatments. The results showed that the rate of anomalous pupae in exposed honeybees was statistically significant in relationship to the control (p <0:03). The most frequent abnormalities were: high pigmentation on the proximal and distal larval body and body malformation, such as absence of head and limbs. Pink eye pupa and white eyed pupae presented malformations in their larval bodies, but with the eye developed. It is assumed that the fat body is related to the high rate of anomalies, since this tissue has proteins linked to the process of metamorphosis. Furthermore, the fat body may be participating in the regulation of juvenile hormone during the process of metamorphosis, and consequently in the release of ecdysteroid hormones that are involved in the change from larva to adult. The high rate of abnormalities in the pupal stage of individuals exposed to fipronil raises concerns about the impacts caused in the colonies of bees and population decline of pollinators. Keywords: bees, larvae, pupae, metamorphosis, anomalies, fiproni

Losses of Brazilian bees: an overview of factors that may affect these pollinators

The Neotropical region to which Brazil belongs, has a great and rich diversity of natives bees, up to a total of 3.000 species including the allochtone genus Apis that by natural crossing among European and African races produced a hybrid called Africanized honeybee. In this way, beekeeping enjoys a spectacular moment with good production mainly of honey and propolis from Apis mellifera, causing Brazil to be recognized around the world as a country with great potential. Brazilian bee losses nevertheless remain a question, compared with countries of the northern hemisphere where several reports show that the vanishing of honeybees is associated with diseases caused by Varroa, Nosema, virus or pesticides. We can suggest different issues in the Brazilian situation that are directly influencing the honeybee population. Given the extension of the territory and rich flora, all possible food resources and nest sites for the good development of bees may be considered present. However, we verified that annual bee losses in the Southeast can reach 20-30%, mainly due the genetic mechanisms of swarming (nest abandon). Many times the major factor leading to nest abandon is lack of food, often taken mistakenly by the untrained beekeeper as death of the hive caused by diseases or pesticides. Although in Brazil diseases do not represent an important problem for Africanized honeybees, some cases of presence of Nosema ceranae and Varroa destructor led the specialists to precaution and monitoring the colonies. In spite of this, the Brazilian beekeeping is managed without use of any acaricide or antibiotic, producing contaminant-free products. As to pesticides, Brazil has a particular climatic and soil condition that might differently affect the risk of exposure of bees to xenobiotics. For example, comparing the dynamics of carbamate pesticides in soil between Brazil and Europe, it was found that in our condition ten-fold more time is needed to obtain the metabolites sulfone and sulfoxide, both more soluble and toxic than it precursor. Comparing the pesticides consumption, currently Brazil has become the world leader followed by USA, with a total spending of 44.9% herbicide, 28.5% insecticide and 22.1% fungicide. Even with this consumption Brazil still belongs to the group that uses a relatively small amount of active ingredient per hectare, less than Japan and France. However cultures like tomato, potato, citrus, cotton and coffee that are often visited by bees during bloom, are also those where the use of pesticides is needed for the pest control. Thus, little is known yet about pesticide losses of the Brazilian bees! What are real effects of pesticides, toxic plants, diseases, genetic improvement, beekeeping management, starvation or interactions among these? Therefore, our local group on ecotoxicological assessment is trying to increase the knowledge on the pesticides hazard to bees (Apis and non-Apis) in order to protect these. Keywords: Brazilian bees, overview, pesticide, diseases, environment, protection

Novel findings on the impact of chytridiomycosis on the cardiac function of anurans: sensitive vs. tolerant species

Background Understanding of the physiological effects of chytridiomycosis is crucial to worldwide amphibian conservation. Therefore, we analyzed the cardiac function of two anuran species (Xenopus laevis and Physalaemus albonotatus) with different susceptibilities to infection by the causative agent of chytridiomycosis, Batrachochytrium dendrobatidis (hereafter Bd). Methods We analyzed the in situ heart rate (fH - bpm), relative ventricular mass (RVM -%), and Ca2+ handling in heart of Bd infected animals compared to uninfected controls of both study species. Results Bd infection resulted in a 78% decrease in contraction force values in P. albonotatus when compared to the less susceptible X. laevis. This negative effect was even more evident (82%) for the cardiac pumping capacity. The time to reach peak tension was 125% longer in P. albonotatus than in X. laevis, and cardiac relaxation was 57% longer. Discussion These results indicate a delay in the cardiac cycle of P. albonotatus on a beat-to-beat basis, which was corroborated by the bradycardia observed in situ. In summary, Bd-sensitive species present impaired cardiac function, which could be a factor in mortality risk. The more pronounced effects of Bd in P. albonotatus may not only result from electrolyte imbalance, as previously reported, but also could be an effect of toxins produced by Bd. For X. laevis, the ability to promote cardiac adjustments seems to be an important homeostatic feature that allows greater tolerance to chytridiomycosis. This study provides new physiological mechanisms underlying the tolerance or susceptibility of amphibian species to chytridiomycosis, which determine their adaptability to survive in the affected environments

In situ localization of heat-shock proteins and cell death labelling in the salivary gland of acaricide-treated honeybee larvae

The effects of the acaricides, rotenone and oxalic acid (OA) on salivary glands of honeybee larvae were evaluated. Immunohistochemical methods were used to detect cell death and heat-shock protein (HSP70 and 90) localizations. Heat-shock proteins (HSP70 and 90) were localized in the cytoplasm and/or the nuclei of secretory gland cells, both under stress and in normal conditions. In rotenone-treated larvae, there were no changes in the normal level of cell death and also there were no morphological alterations in the secretory cells. In the larvae treated with oxalic acid, the salivary gland showed varying degrees of morphological cellular alteration and an increase in the cell death level. The present data suggest that stress-induced HSP70 might have an antiapoptotic effect while the stress-induced HSP90 might have a chaperone function in the larval salivary glands

Morphological alterations induced by boric acid and fipronil in the midgut of worker honeybee (Apis mellifera L.) larvae

Morphological alterations, by means of histological and ultrastructural analysis, have been used to determine the effects of boric acid and fipronil on midgut tissues of honeybee worker, Apis mellifera L. larvae. In order to observe possible morphological alterations in the midgut, two groups of bioassays were performed. In the first one, the larvae were chronically treated with different concentrations of boric acid added to the food (1.0, 2.5 and 7.5 mg/g). In the second group, the larvae were fed with diets containing different concentrations of fipronil (0.1 and 1 mu g/g) and compared with control groups without these chemical compounds. In the first bioassay, the larvae were collected on day 3 and in the second bioassay on day 4, when the mortality rate obtained in the toxicological bioassay was not very high. The larval midguts were removed and processed for morphological analyses using a light and transmission electron microscopy. We observed cytoplasmic vacuolizations, with the absence of autophagic vacuoles, and chromatinic compacting in most of the cells in the groups treated with pesticides. The morphological alterations were far greater in the larvae treated with boric acid than in the larvae treated with fipronil. Our data suggest that the midgut cell death observed was in response to boric acid and fipronil action. This study significantly improves the understanding of the toxicological effect of these insecticides from the ecotoxicological perspective.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Harmful Effects of Pyraclostrobin on the Fat Body and Pericardial Cells of Foragers of Africanized Honey Bee

Managed honey bees are daily exposed in agricultural settings or wild environments to multiple stressors. Currently, fungicide residues are increasingly present in bees’ pollen and nectar and can harm colonies’ production and survival. Therefore, our study aimed to evaluate the effects of the fungicide pyraclostrobin on the fat body and pericardial cells of Africanized honey bees. The foragers were divided into three experimental treatment groups and two controls: pyraclostrobin 0.125 ng/µL (FG1), 0.025 ng/µL (FG2), 0.005 ng/µL (FG3), untreated control (CTL), and acetone control (CAC). After five days of oral exposure (ad libitum), the bees were dissected and prepared for histopathological and morphometric analysis. The FG1-treated bees showed extensive cytoarchitecture changes in the fat body and pericardial cells, inducing cell death. Bees from the FG2 group showed disarranged oenocytes, peripheral vacuolization, and pyknotic nuclei of pericardial cells, but the cytoarchitecture was not compromised as observed in FG1. Additionally, immune system cells were observed through the fat body in the FG1 group. Bees exposed to FG3 demonstrated only oenocytes vacuolization. A significant decrease in the oenocyte’s surface area for bees exposed to all pyraclostrobin concentrations was observed compared to the CTL and CAC groups. The bees from the FG1 and FG2 treatment groups presented a reduced surface area of pericardial cells compared to the controls and the FG3 group. This study highlighted the harmful effects of fungicide pyraclostrobin concentrations at the individual bee cellular level, potentially harming the colony level on continuous exposure

Toxicological and histopathological effects of boric acid on atta sexdens rubropilosa (Hymenoptera: Formicidae) workers

The current study compared the toxicity of different concentrations of boric acid in adult workers of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae), with toxicological bioassays, and examining the dose-dependent and time-dependent histopathological changes, of the midgut, Malpighian tubules, and postpharyngeal glands. Our results revealed the importance of conducting toxicological bioassays combined with morphological analyses of the organs of ants chronically exposed to insecticides used in commercial ant baits. In vitro bioassays showed that boric acid significantly decreases the survivorship of workers regardless of concentration, whereas the morphological data suggested progressive dose-dependent and time-dependent changes in the organs examined, which were evident in the midgut. The midgut is the first organ to be affected, followed by the postpharyngeal gland and Malpighian tubules. This sequence is in agreement with the absorption pathway of this chemical compound in the midgut, its transference to the hemolymph, possibly reaching the postpharyngeal glands, and excretion by the Malpighian tubules. These progressive changes might be due to the cumulative and delayed effect of boric acid. Our findings provide important information for the understanding of the action of boric acid in ant baits in direct and indirect target organs. © 2010 Entomological Society of America