Effects of sublethal doses of imidacloprid in malpighian tubules of africanized Apis mellifera (Hymenoptera, Apidae)

In Brazil, imidacloprid is a widely used insecticide on agriculture and can harm bees, which are important pollinators. The active ingredient imidacloprid has action on the nervous system of the insects. However, little has been studied about the actions of the insecticide on nontarget organs of insects, such as the Malpighian tubules that make up the excretory and osmoregulatory system. Hence, in this study, we evaluated the effects of chronic exposure to sublethal doses of imidacloprid in Malpighian tubules of Africanized Apis mellifera. In the tubules of treated bees, we found an increase in the number of cells with picnotic nuclei, the lost of part of the cell into the lumen, and a homogenization of coloring cytoplasm. Furthermore, we observed the presence of cytoplasmic vacuolization. We confirmed the increased occurrence of picnotic nuclei by using the Feulgan reaction, which showed the chromatin compaction was more intense in the tubules of bees exposed to the insecticide. We observed an intensification of the staining of the nucleus with Xylidine Ponceau, further verifying the cytoplasmic negative regions that may indicate autophagic activity. Additionally, immunocytochemistry experiments showed TUNEL positive nuclei in exposed bees, implicating increased cell apoptosis after chronic imidacloprid exposure. In conclusion, our results indicate that very low concentrations of imidacloprid lead to cytotoxic activity in the Malpighian tubules of exposed bees at all tested times for exposure and imply that this insecticide can alter honey bee physiology. © 2013 Wiley Periodicals, Inc.

Brain morphophysiology of Africanized bee Apis mellifera exposed to sublethal doses of imidacloprid

Several synthetic substances are used in agricultural areas to combat insect pests; however, the indiscriminate use of these products may affect nontarget insects, such as bees. In Brazil, one of the most widely used insecticides is imidacloprid, which targets the nervous system of insects. Therefore, the aim of this study was to evaluate the effects of chronic exposure to sublethal doses of imidacloprid on the brain of the Africanized Apis mellifera. The organs of both control bees and bees exposed to insecticide were subjected to morphological, histochemical and immunocytochemical analysis after exposure to imidacloprid, respectively, for 1, 3, 5, 7, and 10 days. In mushroom bodies of bees exposed to imidacloprid concentrations of LD50/10 and in optic lobes of bees exposed to imidacloprid concentrations of LD 50/10, LD50/100, and LD50/50, we observed the presence of condensed cells. The Feulgen reaction revealed the presence of some cells with pyknotic nuclei, whereas Xylidine Ponceau stain revealed strongly stained cells. These characteristics can indicate the occurrence of cell death. Furthermore, cells in mushroom bodies of bees exposed to imidacloprid concentrations of LD50/10 appeared to be swollen. Cell death was confirmed by immunocytochemical technique. Therefore, it was concluded that sublethal doses of imidacloprid have cytotoxic effects on exposed bee brains and that optic lobes are more sensitive to the insecticide than other regions of the brain. © 2013 Springer Science+Business Media New York.

Effect of sublethal doses of the insecticide imidacloprid on adaptive traits of Drosophila melanogaster: Response to treatment over and after 15 consecutive generations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

FIPRONIL promotes motor and behavioral changes in honey bees (APIS Mellifera) and affects the development of colonies exposed to sublethal doses

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of sublethal doses of methoxyfenozide on the development, survival and reproduction of Spodoptera frugiperda.

The lethal and sublethal effects of the ecdysone agonist methoxyfenozide on the fall armyworm, Spodoptera frugiperda (J. E. Smith), were investigated by feeding a methoxyfenozide-treated diet to ϐifth instars until pupation in doses corresponding to the LC 10 and LC 25 for the compound. Larval mortality reached 8% and 26% in the low and high concentration groups, respectively, on the seventh day of the experiment. A progressive larval mortality of 12% for the LC 10 and 60% for the LC 25 was observed before pupation. Treated larvae exhibited lower pupal weights, higher pupal mortality, presence of deformed pupae, and more deformed adults than untreated larvae. The incorporation of methoxyfenozide into the diet had a signiϐicant effect on the timing of larval development. The development period for males and females was about seven days longer than the controls for both concentrations tested. In contrast, the compound affected neither pupae nor adult longevity. Finally, S. frugiperda adults that resulted from ϐifth instars treated with methoxyfenozide were not affected in their mean cumulative number of eggs laid per female (fecundity), nor percentages of eggs hatched (fertility), or the sex ratio. Our results suggest that the combination of lethal and sublethal effects of methoxyfenozide may have important implications for the population dynamics of the fall armyworm

Germinação de sementes de guandu sob efeito da disponibilidade hídrica e de doses subletais de alumínio

O objetivo deste trabalho foi verificar a germinação de sementes das cultivares de guandu, IAPAR 43-Aratã e IAC Fava Larga, sob efeito da disponibilidade hídrica e de doses subletais de alumínio. Os experimentos foram instalados na Faculdade de Ciências Agrárias e Veterinárias-UNESP, em Jaboticabal (SP), no período de março a maio de 2002. As sementes foram colocadas em caixas de germinação, previamente esterilizadas, revestidas com uma folha de papel germitest umedecido com diferentes concentrações de PEG 6000 (0,0; -0,6; -0,9; -1,2 e -1,5 MPa) e de sulfato de alumínio (0,0; 2,5; 5,0; 7,5; e 10,0 mmol.dm-3). O delineamento experimental utilizado foi o inteiramente casualizado em arranjo fatorial 2 x 5 x 5 (cultivares x disponibilidades hídricas x alumínio), com quatro repetições de 50 sementes. Para complementar a análise estatística foram realizadas análises multivariadas de agrupamento e componentes principais. O experimento foi desenvolvido em câmara de germinação, na temperatura de 25 ºC. A porcentagem de germinação foi verificada no 4.º e 10.º dias após a semeadura. No 10.º dia, as plântulas foram separadas em plúmulas e raízes primárias, para determinar a massa seca. A disponibilidade hídrica foi limitante para a germinação e o crescimento inicial. O efeito do alumínio associado ao estresse hídrico foi evidenciado somente até -0,9 MPa. As concentrações menos elevadas de alumínio estimularam o desenvolvimento das plântulas em condições de estresse hídrico até -0,6 MPa. Os efeitos simultâneos da disponibilidade hídrica e do alumínio reduziram a germinação de sementes da cv. IAC Fava Larga e o crescimento das raízes da cv. IAPAR 43-Aratã.

Biochemical and proteomic effects in Procambarus clarkii after chlorpyrifos or carbaryl exposure under sublethal conditions

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeitos de doses subletais do imidaclopride no cérebro, ventrículo e túbulo de Malpighi de Apis melifera africanizada

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeitos das doses subletais de fipronil para abelhas africanizadas (Apis mellifera L.), por meio de análises morfológicas e comportamentais

Pós-graduação em Ciências Biológicas (Biologia Celular e Molecular) - IBRC

Sublethal effects of a common neonicotinoid pesticide, thiamethoxam, on honey bees: impact on locomotion and thermoregulation

Neonicotinoids have been pointed to as a factor responsible for the increased honey bee colony losses in the last decades. Many studies have investigated the effects of the first marketed neonicotinoid, imidacloprid, while fewer have focused on thiamethoxam. One recent study showed that sublethal doses of thiamethoxam lead to colony failure by decreasing forager homing flight success. We thus decided to investigate the mechanism which caused this phenomenon. Our hypothesis was that this effect was caused by impairment of forager locomotion abilities. Therefore we tested the effects of sublethal acute and chronic exposures to thiamethoxam on forager walking (Chapter 2) and flight (Chapter 3) performances. The acute treatment (1.34 ng/bee) affected walking locomotion firstly triggering hyperactivity (30 min post-treatment) and then impairing motor functioning (60 min post-treatment). 2-day continuous exposures to thiamethoxam (32.5, 45 ppb) elicited fewer effects on walking locomotion, however both exposure modes elicited an increased positive phototaxis. Similarly, in flight experiments, the single dose (1.34 ng/bee) elicited hyperactivity shortly after intoxication (increased flight duration and distance), while longer and continuous exposures (32.5, 45 ppb) impaired forager motor functions (decreased flight duration, distance, velocity). It is known that flight muscles temperature needs to be precisely regulated by bees during flight. Therefore, we further hypothesized that the impaired flight performances of neonicotinoid intoxicated bees were caused also by thermoregulation anomalies. We tested the effects that acute thiamethoxam exposures (0.2, 1, 2 ng/bee) elicit on forager thorax temperature (Chapter 4). Foragers treated with high doses exhibited hyperthermia or hypothermia when respectively exposed to high or low environmental temperatures. In summary, we show that sublethal doses of thiamethoxam affected forager walking and flight locomotion, phototaxis and thermoregulation. We also display the intricate mode of action of thiamethoxam which triggered, at different extents, inverse sublethal effects in relation to time and dose.

Radiation metabolomics. 2. Dose- and time-dependent urinary excretion of deaminated purines and pyrimidines after sublethal gamma-radiation exposure in mice

Gamma-radiation exposure of humans is a major public health concern as the threat of terrorism and potential hostile use of radiological devices increases worldwide. We report here the effects of sublethal gamma-radiation exposure on the mouse urinary metabolome determined using ultra-performance liquid chromatography-coupled time-of-flight mass spectrometry-based metabolomics. Five urinary biomarkers of sublethal radiation exposure that were statistically significantly elevated during the first 24 h after exposure to doses ranging from 1 to 3 Gy were unequivocally identified by tandem mass spectrometry. These are deaminated purine and pyrimidine derivatives, namely, thymidine, 2'-deoxyuridine, 2'-deoxyxanthosine, xanthine and xanthosine. Furthermore, the aminopyrimidine 2'-deoxycytidine appeared to display reduced urinary excretion at 2 and 3 Gy. The elevated biomarkers displayed a time-dependent excretion, peaking in urine at 8-12 h but returning to baseline by 36 h after exposure. It is proposed that 2'-deoxyuridine and 2'-deoxyxanthosine arise as a result of gamma irradiation by nitrosative deamination of 2'-deoxycytidine and 2'-deoxyguanosine, respectively, and that this further leads to increased synthesis of thymidine, xanthine and xanthosine. The urinary excretion of deaminated purines and pyrimidines, at the expense of aminopurines and aminopyrimidines, appears to form the core of the urinary radiation metabolomic signature of mice exposed to sublethal doses of ionizing radiation.

Pre-treatment of Stegomyia aegypti mosquitoes with a sublethal dose of imidacloprid impairs behavioural avoidance induced by lemon oil and DEET

International audience

Low doses of a neonicotinoid insecticide modify pheromone response thresholds of central but not peripheral olfactory neurons in a pest insect

International audience

Environmental poisoning: Presentation and management

Environmental poisoning is most commonly associated with chronic longterm exposure to toxins rather than to acute exposure. Such repeated exposure to sublethal doses of compounds and elements presents problems in risk assessment. This is primarily because the data are unavailable to describe relationships between dose and effect at lower levels of exposure to toxins. Bioavailability of toxins also presents a problem because the data on bioavailability are sparse and seldom as high as the default of 100% bioavailability commonly used in risk assessment. Examples are presented of two toxins: arsenic as an elemental anthropogenic and geologic poison and ciguatoxin, a polyether ladder compound, as a toxin produced naturally by dinoflagellates. Bioavailability drives the toxicity of arsenic from contaminated sites, whereas tissue accumulation drives the toxicity of ciguatoxin. Considerable benefit is derived from the harmonization of regulatory processes where there is linkage of health and environmental factors in the derivation of credible risk assessment.

Evaluation of synergistic interactions between the Colorado potato beetle (Coleoptera : Chrysomelidae) pathogen Beauveria bassiana and the insecticides, imidacloprid, and cyromazine

Laboratory studies investigated the interaction between the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin and sublethal doses of the insecticides imidacloprid and cyromazine when applied to larvae of the Colorado potato beetle, Leptinotarsa decemlinenta (Say). When second instars were fed potato leaf discs treated with sublethal doses of imidacloprid and a range of doses of B. bassiana, a synergistic action was demonstrated. Similar results were observed when larvae were sprayed directly with B. bassiana conidia and immediately fed leaf discs treated with imidacloprid. No synergistic interaction was detected when larvae were fed leaf discs treated with sublethal doses of imidacloprid 24 h after application of R. bassiana conidia to larvae. However, a synergistic interaction was detected when larvae were fed leaf discs treated with imidacloprid and sprayed with B, bassiana conidia 24 h later. Although sublethal doses of both imidacloprid and the triazine insect growth regulator (IGR) cyromazine prolonged the duration of the second instar, only imidacloprid interacted with B. bassiana to produce a synergistic response in larval mortality. In leaf consumption studies, the highest dose of B, bassiana tested promoted feeding in inoculated second instars. Feeding was inhibited when larvae were fed foliage treated with sublethal doses of imidacloprid and significantly reduced when fed foliage treated with a sublethal dose of cyromazine. Starvation of larvae for 24 h immediately after B. bassiana treatment produced a similar result to the combined treatment of B. bassiana and imidacloprid and increased the level of mycosis when compared with B. bassiana controls. Imidacloprid treatment affected neither the rate of germination of B. bassiana conidia on the insect cuticle nor the rate at which conidia were removed from the integument after application. The statistical analysis used to detect synergism and the possible role of starvation-induced stress factors underlying the observed synergistic interactions are discussed.