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Wheat production is dependent on costly insecticides that generate social and environmental issues. The growing demand for a rate reduction of spray applications and increased spray efficiency make essential the research for more adequate crop management. Therefore, this study aimed to evaluate technologies of imidacloprid application on wheat aphid, Schizaphis graminum and its natural enemies, Chrysoperla externa and Orius insidiosus. A set of experiments were performed to evaluate the effectiveness of S. graminum control and the toxicity to natural enemies using imidacloprid plus adjuvants sprayed with different nozzles and application rates. Improved S. graminum control was observed in the association between the adjuvant lauryl ether sodium sulfate and the hollow cone nozzle for both application rates (75 and 150 L ha-1), and the adjuvant copolymer of polyester + silicone + d-limonene with the asymmetric twin flat-fan nozzle at a low rate (75 L ha-1), since these treatments eliminate the wheat aphid after the fifth day of treatment application. The reduction of spray volume to wheat aphid control is possible with no loss of insecticide effectiveness. Imidacloprid associated with any one of the adjuvants tested is harmful to C. externa and O. insidiosus, even at the fifth day after application the insecticide continues being harmful to the natural enemies, regardless of the technology used. Key words: Triticum aestivum, Schizaphis graminum, ecological disturbance, insecticide impacts, natural control, neonicotinoid.

Thiamethoxam (TMX) belongs to a class of neuro-active insecticides referred as neonicotinoids, while actara® (AC) is one of the most popular TMX-based products in Brazil. The aim of this study was to evaluate the mutagenic, recombinogenic and carcinogenic potential of TMX and AC insecticides. The mutagenic and recombinogenic effect of TMX and AC were evaluated in vivo by the Somatic Mutation and Recombination Test (SMART) while carcinogenic effects were evaluated through the Test for Detection of Epithelial Tumor Clones (wts test), both in somatic cells of Drosophila melanogaster. In the SMART, third instar larvae from standard (ST) and high bioactivation (HB) crosses were treated with different concentrations of TMX and AC (2.4; 4.8; 9.7 × 10−4 mM and 1.9 × 10−3 mM). The results revealed mutagenic effects at the highest concentrations tested in the HB cross. In the test for the detection of epithelial tumor, third instar larvae resulting from the cross between wts/TM3, Sb1 virgin females and mwh/mwh males were treated with the same concentrations of TMX and AC used in the SMART. No carcinogenic effect was observed at any of the concentrations tested. In this work, the inhibition of the mechanism of repair by homologous recombination was observed in flies exposed to 9.7 × 10−4 and 1.9 × 10−3 mM of AC. In conclusion, TMX and AC demonstrated to be a promutagen in the highest concentrations tested.

Bees are recognized worldwide for their social, economic, and environmental value. In recent decades they have been seriously threatened by diseases and high levels of pesticide use. The susceptibility of bees to insecticides makes them an important terrestrial model for assessing environmental quality, and various biomarkers have been developed for such assessments. The present study aimed to evaluate the activity of the enzymes acetylcholinesterase (AChE), carboxylesterase (CaE), and glutathione-S-transferase (GST) in Africanized honeybees exposed to fipronil. The results showed that fipronil at a sublethal dose (0.01 ng/bee) modulates the activity of CaE in all isoforms analyzed (CaE-1, CaE-2, and CaE-3) in both newly emerged and aged bees, and does not affect the activity of AChE or GST. The recovery of the bees after fipronil exposure was also investigated, and these results demonstrated that even the cessation of fipronil ingestion might not lead to complete recovery of individual bees. Even at low doses, fipronil was shown to cause changes in the activity of key enzymes in bees. The possible consequences of these changes are discussed. Environ Toxicol Chem 2017;36:1652-1660. © 2016 SETAC.

This study used the pearl cichlid Geophagus brasiliensis as a bioindicator to survey the health of the aquatic environment on four sites (P1, P2, P3 and P4) of the Mumbuca stream located at Monte Carmelo/MG, Brazil. The selection of different sites was made with reference to the gradient of urban activity and via physicochemical and biological evaluation of water quality and genotoxicity. The water quality index was classified as 'good' for P1 and P4, regular in P2 and 'poor' for P3. The micronuclei (MN) frequency obtained from blood analysis was in agreement with the water quality, such that the higher values of MN were detected in sites evaluated as poor. Water degradation conditions worsen according to the flow of the stream over the sites P1, P2 and P3, but for site P4, located after the Monte Carmelo Sewage Treatment Plant, improvements in the micronuclei frequency are detected. Our results showed high levels of potentially toxic metals (chromium, lead, aluminum and nickel) in specific stream sites (P2 and P3). We suggest that the micronuclei induction in G. brasiliensis could be due to the presence of these compounds.

Made available in DSpace on 2018-11-26T16:16:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Several investigations have revealed the toxic effects that neonicotinoids can have on Apis mellifera, while few studies have evaluated the impact of these insecticides can have on the larval stage of the honeybee. From the lethal concentration (LC50) of thiamethoxam for the larvae of the Africanized honeybee, we evaluated the sublethal effects of this insecticide on morphology of the brain. After determine the LC50 (14.34 ng/mu L of diet) of thiamethoxam, larvae were exposed to a sublethal concentration of thiamethoxam equivalent to 1.43 ng/mu L by acute and subchronic exposure. Morphological and immunocytochemistry analysis of the brains of the exposed bees, showed condensed cells and early cell death in the optic lobes. Additional dose-related effects were observed on larval development. Our results show that the sublethal concentrations of thiamethoxam tested are toxic to Africanized honeybees larvae and can modulate the development and consequently could affect the maintenance and survival of the colony. These results represent the first assessment of the effects of thiamethoxam in Africanized honeybee larvae and should contribute to studies on honey bee colony decline. (C) 2015 Elsevier Ltd. All rights reserved. Univ Estadual Paulista, Dept Biol, BR-13506900 Rio Claro, SP, Brazil Univ Fed Uberlandia, Inst Ciencias Agr, BR-38400 Uberlandia, MG, Brazil Univ Fed Sao Carlos, Dept Biol, Sorocaba, SP, Brazil Univ Estadual Paulista, Dept Biol, BR-13506900 Rio Claro, SP, Brazil FAPESP: 2009/06701-5 FAPESP: 2013/09419-4 FAPESP: 2012/50197-2

The stingless bee Scaptotrigona postica is an important pollinator of native and cultivated plants in Brazil. Among the factors affecting the survival of these insects is the indiscriminate use of insecticides, including the neonicotinoid imidacloprid. This work determined the toxicity of imidacloprid as the topical median lethal dose (LD50) and the oral median lethal concentration (LC50) as tools for assessing the effects of this insecticide. The 24 and 48 h LD50 values were 25.2 and 24.5 ng of active ingredient (a.i.)/bee, respectively. The 24 and 48 h LC50 values were 42.5 and 14.3 ng a.i./µL of diet, respectively. Ours results show the hazard of imidacloprid and the vulnerability of stingless bees to it, providing relevant toxicological data that can used in mitigation programs to ensure the conservation of this species.

Made available in DSpace on 2018-12-11T17:32:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Under laboratory conditions, the effects of thiamethoxam were investigated in larvae, pupae and emerging honey bees after exposure at larval stages with different concentrations in the food (0.00001 ng/μL, 0.001 ng/μL and 1.44 ng/μL). Thiamethoxam reduced the survival of larvae and pupae and consequently decreased the percentage of emerging honey bees. Thiamethoxam induced important physiological disturbances. It increased acetylcholinesterase (AChE) activity at all developmental stages and increased glutathione-S-transferase (GST) and carboxylesterase para (CaEp) activities at the pupal stages. For midgut alkaline phosphatase (ALP), no activity was detected in pupae stages, and no effect was observed in larvae and emerging bees. We assume that the effects of thiamethoxam on the survival, emergence and physiology of honey bees may affect the development of the colony. These results showed that attention should be paid to the exposure to pesticides during the developmental stages of the honey bee. This study represents the first investigation of the effects of thiamethoxam on the development of A. mellifera following larval exposure. UNESP Universidade Estadual Paulista Departamento de Biologia INRA Laboratoire de Toxicologie Environnementale UR 406 Abeilles & Environnement INRA UR 546 Biostatistique et Processus Spatiaux UFLA Universidade Federal de Lavras, Lavras UFSCar Universidade Federal de São Carlos INRA Laboratoire de L'Environnement et de L'Alimentation de La Vendée, La Roche sur Yon UNESP Universidade Estadual Paulista Departamento de Biologia FAPESP: 2013/21634-8

The development of agricultural activities coincides with the increased use of pesticides to control pests, which can also be harmful to nontarget insects such as bees. Thus, the goal of this work was assess the toxic effects of thiamethoxam on newly emerged worker bees of Apis mellifera (africanized honeybee—AHB). Initially, we determined that the lethal concentration 50 (LC50) of thiamethoxam was 4.28 ng a.i./lL of diet. To determine the lethal time 50 (LT50), a survival assay was conducted using diets containing sublethal doses of thiamethoxam equal to 1/10 and 1/100 of the LC50. The group of bees exposed to 1/10 of the LC50 had a 41.2% reduction of lifespan. When AHB samples were analyzed by morphological technique we found the presence of condensed cells in the mushroom bodies and optical lobes in exposed honeybees. Through Xylidine Ponceau technique, we found cells which stained more intensely in groups exposed to thiamethoxam. The digestive and regenerative cells of the midgut from exposed bees also showed morphological and histochemical alterations, like cytoplasm vacuolization, increased apocrine secretion and increased cell elimination. Thus, intoxication with a sublethal doses of thiamethoxam can cause impairment in the brain and midgut of AHB and contribute to the honeybee life

Fipronil is an insecticide widely used to control a great number of pests, thus the aim of this study was to determine the lethal dose and lethal concentration (LD(50) and LC(50)) of this insecticide to the stingless bees Scaptotrigona postica Latreille, 1807. The LD(50) and LC(50) values obtained after 24 h of exposition were of 0.54 ng a.i./bee and 0.24 ng a.i./μL diet, respectively. These values were considered highly toxic to stingless bees.

TOXICITY OF ACARICIDES/INSECTICIDES FOR CITRUS CROP TO THE AFRICANIZED HONEYBEE APIS MELLIFERA L., 1758 (HYMENOPTERA: APIDAE). The purpose of this work was to evaluate the toxicity of commercial pesticides used on citrus crop to the honey bees Apis mellifera Linnaeus. The number of dead honey bees was registered during time assays, which were carried out in laboratory conditions with the chemicals thiamethoxam, deltamethrin, lufenuron, tebufenozide, propargite, cyhexatin, methidathion and abamectin. The exposure of the honey bees to the chemicals was made by spraying, food contamination and contact with contaminated surfaces. In all assays, thiamethoxam, methidathion and abamectin were highly toxic, with average lethal times (LT 50 ) of 3.57; 3.34 and 23.12h, respectively. In spraying assays, deltamethrin showed low toxicity to the honey bees, however, in supplied food contamination and residue contact tests, this pyrethroid was very toxic. For propargite, medium toxicity was verified when the contaminated food was supplied to the bees, with a LT 50 of 64.65h, and in the other assays, a similar effect to cyhexatin, tebufenozide and lufenuron was verified, with low toxicity to the adult of africanized honeybees.

Aquatic organisms are considered excellent biomarkers of mercury (Hg) occurrence in the environment. Selenium (Se) acts in antagonism to this metal, stimulating its elimination, and reducing its toxicity. In this paper, tilapia (Oreochromis niloticus) were chronically acclimated in sub-lethal Hg(2+), Hg(2+) + Se(4+) and Hg(2+) + Se(6+) concentrations. Distribution and bioaccumulation of both elements were evaluated in fish tissues. The kidney was the main target of the Hg and Se uptake, and the presence of Hg induced the Se hepatic elimination. The Hg bioaccumulation in the gill, spleen and heart were higher in the presence of Se(6+) than in the presence of Se(4+).

Made available in DSpace on 2018-12-11T17:00:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-03-01 Multiple stressors, such as chemicals and pathogens, are likely to be detrimental for the health and lifespan of Apis mellifera, a bee species frequently exposed to both factors in the field and inside hives. The main objective of the present study was to evaluate comparatively the health of Carniolan and Africanized honey bees (AHB) co-exposed to thiamethoxam and Nosema ceranae (N. ceranae) spores. Newly-emerged worker honey bees were exposed solely with different sublethal doses of thiamethoxam (2% and 0.2% of LD50 for AHB), which could be consumed by bees under field conditions. Toxicity tests for the Carniolan bees were performed, and the LD50 of thiamethoxam for Carniolan honey bees was 7.86 ng bee-1. Immunohistological analyses were also performed to detect cell death in the midgut of thiamethoxam and/or N. ceranae treated bees. Thiamethoxam exposure had no negative impact on Nosema development in experimental conditions, but it clearly inhibited cell death in the midgut of thiamethoxam and Nosema-exposed bees, as demonstrated by immunohistochemical data. Indeed, thiamethoxam exposure only had a minor synergistic toxic effect on midgut tissue when applied as a low dose simultaneously with N. ceranae to AHB and Carniolan honey bees, in comparison with the effect caused by both stressors separately. Our data provides insights into the effects of the neonicotenoid thiamethoxam on the AHB and Carniolan honey bee life span, as well as the effects of simultaneous application of thiamethoxam and N. ceranae spores to honey bees. Agricultural Institute of Slovenia Universidade Federal de São Carlos (UFSCar) Universidade Federal de Uberlândia University of Maribor Faculty of Agriculture and Life Sciences Laboratório de Sanidade Apícola (LASA) Agência Paulista de Tecnologia dos Agronegócios (APTA) Centro de Estudos de Insetos Sociais (CEIS) Instituto de Biociências UNESP, Campus de Rio Claro Centro de Estudos de Insetos Sociais (CEIS) Instituto de Biociências UNESP, Campus de Rio Claro

Fipronil is a neurotoxic insecticide that inhibits the gamma-aminobutyric acid receptor and can affect gustative perception, olfactory learning, and motor activity of the honeybee Apis mellifera. This study determined the lethal dose (LD50) and the lethal concentration (LC50) for Africanized honeybee and evaluated the toxicity of a sublethal dose of fipronil on neuron metabolic activity by way of histochemical analysis using cytochrome oxidase detection in brains from worker bees of different ages. In addition, the present study investigated the recovery mechanism by discontinuing the oral exposure to fipronil. The results showed that mushroom bodies of aged Africanized honeybees are affected by fipronil, which causes changes in metabolism by increasing the respiratory activity of mitochondria. In antennal lobes, the sublethal dose of fipronil did not cause an increase in metabolic activity. The recovery experiments showed that discontinued exposure to a diet contaminated with fipronil did not lead to recovery of neural activity. Our results show that even at very low concentrations, fipronil is harmful to honeybees and can induce several types of injuries to honeybee physiology.

For a better evaluation of the model using Apis mellifera in toxicology studies with insecticides, the oral acute toxicity of the insecticide fipronil against the stingless bee Melipona scutellaris was determined. The results showed that fipronil was highly toxic to M. scutellaris, with a calculated LC(50) (48 h) value of 0.011 ng a.i./μL of sucrose solution and an estimated oral LD(50) (48 h) of 0.6 ng a.i./bee. Our results showed that M. scutellaris bee is more sensitive to fipronil than the model specie A. mellifera.

International audience; This study describes the development of acetylcholinesterase (AChE), carboxylesterases (CaE1, CaE2, CaE3), glutathion-S-transferase (GST), alkaline phosphatase (ALP) and catalase (CAT) as enzyme biomarkers of exposure to xenobiotics such as thiamethoxam in the honey bee Apis mellifera. Extraction efficiency, stability under freezing and biological variability were studied. The extraction procedure achieved good recovery rates in one extraction step and ranged from 65 percent (AChE) to 97.3 percent (GST). Most of the enzymes were stable at −20 °C, except ALP that displayed a slight but progressive decrease in its activity. Modifications of enzyme activities were considered after exposure to thiamethoxam at the lethal dose 50 percent (LD50, 51.16 ng bee−1) and two sublethal doses, LD50/10 (5.12 ng bee−1) and LD50/20 (2.56 ng bee−1). The biomarker responses revealed that, even at the lowest dose used, exposure to thiamethoxam elicited sublethal effects and modified the activity of CaEs, GST, CAT and ALP. Different patterns of biomarker responses were observed: no response for AChE, an increase for GST and CAT, and differential effects for CaEs isoforms with a decrease in CaE1 and CaE3 and an increase in CaE2. ALP and CaE3 displayed contrasting variations but only at 2.56 ng bee−1. We consider that this profile of biomarker variation could represent a useful fingerprint to characterise exposure to thiamethoxam in the honey bee A. mellifera. This battery of honey bee biomarkers might be a promising option to biomonitor the health of aerial and terrestrial ecosystems and to generate valuable information on the modes of action of pesticides.

O objetivo deste trabalho foi avaliar a toxicidade de inseticidas/acaricidas utilizados em cultura de citros para operárias africanizadas de Apis mellifera Linnaeus. A exposição das abelhas aos compostos foi realizada usando-se técnicas de pulverização, contaminação da dieta e contato em superfícies tratadas (folhas de citros e placas de Petri), empregando-se as doses máximas recomendadas para a cultura. Os bioensaios foram realizados em laboratório a 25±2°C, UR 70±10% e fotofase de 12h, sendo os dados de mortalidade submetidos à análise estatística, e as médias comparadas por contraste, obtendo-se grupos de efeitos semelhantes. Independente do modo de exposição, o acefato foi extremamente tóxico, matando mais de 90,0% das abelhas 24h após a aplicação. Os produtos espirodiclofeno e piriproxifem, quando aplicados diretamente sobre as abelhas, causaram níveis de mortalidade de 11,0 e 15,0%, respectivamente; os compostos buprofezina, enxofre e tetradifona apresentaram níveis de mortalidade ainda menores, com média de 5,0% entre eles. Para os ensaios de contaminação de superfície (folhas de citros e placas de Petri) e contaminação de alimento, foram obtidos dois grupos de toxicidade, um grupo somente com acefato e outro, com buprofezina, enxofre, espirodiclofeno, piriproxifem, tetradifona e água. A mortalidade média para esse segundo grupo, após 96h do início da exposição, foi de 31,0; 8,3 e 15,7%, respectivamente, para cada método de contaminação. The aim of this research was to evaluate the toxicity of several acaricides/insecticides used in Brazilian citrus crop to africanized workers of Apis mellifera Linnaeus. The exposition of honey bees to the chemicals was performed by direct spraying, contamination of food, and contact in treated surface (citrus leaves and Petri dishes), using recommended rates of application. The assays were carried out at 25±2°C, RH 70±10%, 12h of photophase and the data was statistically analyzed, with mean values of mortality being compared through cluster analysis. In all assays acephate was highly toxic, with mortality at 24 hours around 90.0%. When spirodiclofen and pyriproxyfen, was sprayed directly into the honeybees, they caused mortality levels of 11.0 and 15.0%, respectively; buprofezin, sulphur and tetradifon were less toxic, with mean mortality of 5.0% among these compounds. For the assays from contamination surface (citrus leaves and Petri dishes) and food, two groups of chemicals with the same toxic effects were observed, one with acephate and other with buprofezin, sulphur, spirodiclofen, pyriproxyfen, tetradifon and water. The average mortality after 96 hours of exposition was 31.0; 8.3 and 15.7%, respectively, for each method of contamination.

For an important and expensive crop such as corn, the resistance of Spodoptera frugiperda J.E. Smith to various pesticides has led to research throughout the world for a potential insecticide from a natural source. For the management of pest resistance, natural compounds associated with synthetic insecticides can be a promising tool because they can reduce the application of the synthetics molecules while maintaining their effectiveness and promoting the control of the pests. Linalool is a potential insecticide that is easily obtained because it is found in high concentrations in the essential oil of Ocimum basilicum L. Therefore, the present study aimed to evaluate the toxicity of this essential oil and its combination with deltamethrin to control S. frugiperda. Through dose response assays, the acute toxicities (LD50) of the essential oil and deltamethrin were estimated. Additionally, the combination of these materials was also assessed, attaining a reduction of 80% of the LD50 of deltamethrin while obtaining the same result as when the pyrethroid was administered alone. From these results, it is expected that the combination of natural compounds and synthetic insecticides will be a promising practice, helping to manage resistance while reducing the environmental impact of toxic compounds.