Análise morfológica e imunocitoquímica do cérebro de abelhas africanizadas Apis mellifera após exposição à doses subletais do inseticida tiametoxam

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

Toxicidade de Lantana camara (Verbenaceae) em operárias de Apis mellifera (Hymenoptera: Apidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cytotoxic Effects of Thiamethoxam in the Midgut and Malpighian Tubules of Africanized Apis mellifera (Hymenoptera: Apidae)

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

Modification of the brain proteome of Africanized honeybees (Apis mellifera) exposed to a subaEurolethal doses of the insecticide fipronil

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

Hyperalgesic and edematogenic effects of Secapin-2, a peptide isolated from Africanized honeybee (Apis mellifera) venom

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

Estrutura e função de peptídeos policatiônicos de Apis mellifera

Os venenos de animais são bastante estudados devido a um interesse industrial e comercial, pois sua composição é rica em moléculas de importância farmacológica. Os peptídeos presentes nesses venenos apresentam características diversas, como antibiose, analgesia, propriedades antiinflamatórias, vasoconstritora, entre outras. Para caracterização dos componentes peptídicos presentes no veneno de Apis mellifera, primeiramente o veneno foi extraído e filtrado para remoção de eventuais impurezas e depois submetido à cromatografia em HPLC com coluna de fase reversa, que gerou um perfil cromatográfico com 29 picos, dos quais os 3 mais abundantes apresentaram características peptídicas, confirmadas por espectrometria de massas e seqüenciamento. Além disso, a cromatografia de íons totais mostrou que esse veneno apresenta 123 compostos peptídicos, alguns com massas relacionadas aos já conhecidos, porém com diversos compostos de massas diferentes dos compostos já descritos na literatura. A partir deste estudo, detectou-se a presença da Melitina como componente peptídico majoritário no veneno de abelha, além da Secapina, que será caracterizada por estudos em andamento

Estudos sobre atividade nociceptiva/antinociceptiva de peptídeos isolados do veneno bruto de abelha Apis mellifera

O veneno bruto de abelha Apis mellifera é composto por uma série de peptídeos, entre eles a melitina, adolapina, procamina A e B, apamina, tertiapina, secapina, peptídeos desgranuladores de mastócitos, entre outros. Estas substâncias apresentam diversos efeitos já estudadas, dentre elas, efeitos antitumorais, atividades hiperalgésicas ou analgésicas, inflamatórias ou anti-inflamatórias. O peptídeo secapina foi descrito há 35 anos atrás e, posteriormente nada mais foi relatado na literatura. Portanto, este é o primeiro trabalho que visa caracterizar o possível efeito hiperalgésico e/ou analgésico, inflamatório e/ou antiinflamatório, bem como analisar alguns dos mecanismos envolvidos nestes fenômenos. Para tanto, para caracterizar os possíveis efeitos hiperalgésicos/analgésicos da secapina, em diversas doses (0,005, 0,35, 1, 2, 10 e 30 μg / 50 μL), foi utilizado o teste de avaliação da sensibilidade dolorosa denominado teste do von Frey eletrônico. Para a caracterização do efeito inflamatório/anti-inflamatório da secapina foi utilizado o paquímetro digital determinando o volume da pata dos animais. Para investigar os possíveis mecanismos envolvidos nestes efeitos, foram realizados tratamentos farmacológicos utilizando indometacina, zileuton e zafirlikaste (inibidores da síntese da via da cicloxigenase, lipoxigenase e antagonista de receptor de leucotrieno, respectivamente). Os resultados mostraram que a secapina, em todas as doses testadas, induziu efeito hiperalgésico a partir dos primeiros 15 minutos após a administração deste peptídeo. Este efeito hiperalgésico persistiu por até 240 minutos nas doses de 0,005 e 0,35 μg/ 50μL e 480 minutos nas doses de 1 e 2 μg/ 50μL. O efeito edematogênico da secapina na dose de 0,35 μg/50 μL foi iniciado nos primeiros 15 minutos após a administração da secapina e persistiu por até 120 minutos, sendo... (Resumo completo, clicar acesso eletrônico abaixo)

Avaliação dos efeitos de concentrações subletais de dimetoato no cérebro e ventrículo de Apis mellifera africanizada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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)

Avaliação bioquímica e imunohistoquímica dos efeitos subletais do tiametoxam sobre Apis mellifera (Hymenoptera, Apidae)

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

Análise metabolômica do cérebro de abelhas (Apis mellifera) submetidas a ensaio de Reflexo de Extensão de Probóscide (REP)

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

Study of visceral antinociceptive potential of bee Apis mellifera venom

Pain is one of the most common reasons for patients to seek medical care. Bee Apis mellifera venom (AMV) has traditionally been used to treat inflammatory diseases and the alleviation of pain. Herein, we aimed to investigate the visceral antinociceptive potential of A. mellifera bee venom and its possible mechanism of action. Acetic acid-induced writhing assay was used in mice to determine the degree of visceral antinociception. Visceral antinociceptive activity was expressed as the reduction in the number of abdominal constrictions. Mice received an intraperitoneal injection of acetic acid after administration of AMV (0.08 or 0.8 mg/kg; intraperitoneally (i.p.)). In mechanistic studies, separate experiments were realized to examine the role of α2-receptors, nitric oxide, calcium channels, K+ATP channel activation, TRPV1 and opioid receptors on the visceral antinociceptive effect of AMV (0.8 mg/kg), using appropriate antagonists, yohimbine (2 mg/kg), L-NG-Nitroarginine methyl ester (L-NAME, 10 mg/kg), verapamil (5 mg/kg), glibenclamide (5 mg/kg), ruthenium red (3 mg/kg) or naloxone (2 mg/kg). AMV presented visceral antinociceptive activity in both doses tested (0.08 and 0.8 mg/Kg). Visceral antinociceptive effect of AMV was resistant to all the antagonists used. Mice showed no significant alterations in locomotion frequency, indicating that the observed antinociception is not a consequence of motor abnormality. Although AMV efficient diminished the acetic acid-evoked pain-related behavior, its mechanism is unclear from this study and future studies are needed to verify how the venom exerts its antinociceptive action.

Genes involved in thoracic exoskeleton formation during the pupal-to-adult molt in a social insect model, Apis mellifera

Background: The insect exoskeleton provides shape, waterproofing, and locomotion via attached somatic muscles. The exoskeleton is renewed during molting, a process regulated by ecdysteroid hormones. The holometabolous pupa transforms into an adult during the imaginal molt, when the epidermis synthe3sizes the definitive exoskeleton that then differentiates progressively. An important issue in insect development concerns how the exoskeletal regions are constructed to provide their morphological, physiological and mechanical functions. We used whole-genome oligonucleotide microarrays to screen for genes involved in exoskeletal formation in the honeybee thoracic dorsum. Our analysis included three sampling times during the pupal-to-adult molt, i.e., before, during and after the ecdysteroid-induced apolysis that triggers synthesis of the adult exoskeleton. Results: Gene ontology annotation based on orthologous relationships with Drosophila melanogaster genes placed the honeybee differentially expressed genes (DEGs) into distinct categories of Biological Process and Molecular Function, depending on developmental time, revealing the functional elements required for adult exoskeleton formation. Of the 1,253 unique DEGs, 547 were upregulated in the thoracic dorsum after apolysis, suggesting induction by the ecdysteroid pulse. The upregulated gene set included 20 of the 47 cuticular protein (CP) genes that were previously identified in the honeybee genome, and three novel putative CP genes that do not belong to a known CP family. In situ hybridization showed that two of the novel genes were abundantly expressed in the epidermis during adult exoskeleton formation, strongly implicating them as genuine CP genes. Conserved sequence motifs identified the CP genes as members of the CPR, Tweedle, Apidermin, CPF, CPLCP1 and Analogous-to-Peritrophins families. Furthermore, 28 of the 36 muscle-related DEGs were upregulated during the de novo formation of striated fibers attached to the exoskeleton. A search for cis-regulatory motifs in the 5′-untranslated region of the DEGs revealed potential binding sites for known transcription factors. Construction of a regulatory network showed that various upregulated CP- and muscle-related genes (15 and 21 genes, respectively) share common elements, suggesting co-regulation during thoracic exoskeleton formation. Conclusions: These findings help reveal molecular aspects of rigid thoracic exoskeleton formation during the ecdysteroid-coordinated pupal-to-adult molt in the honeybee.

Honey Bee Apis mellifera Parasites in the Absence of Nosema ceranae Fungi and Varroa destructor Mites

Few areas of the world have western honey bee (Apis mellifera) colonies that are free of invasive parasites Nosema ceranae (fungi) and Varroa destructor (mites). Particularly detrimental is V. destructor; in addition to feeding on host haemolymph, these mites are important vectors of several viruses that are further implicated as contributors to honey bee mortality around the world. Thus, the biogeography and attendant consequences of viral communities in the absence of V. destructor are of significant interest. The island of Newfoundland, Province of Newfoundland and Labrador, Canada, is free of V. destructor; the absence of N. ceranae has not been confirmed. Of 55 Newfoundland colonies inspected visually for their strength and six signs of disease, only K-wing had prevalence above 5% (40/55 colonies = 72.7%). Similar to an earlier study, screenings again confirmed the absence of V. destructor, small hive beetles Aethina tumida (Murray), tracheal mites Acarapis woodi (Rennie), and Tropilaelaps spp. ectoparasitic mites. Of a subset of 23 colonies screened molecularly for viruses, none had Israeli acute paralysis virus, Kashmir bee virus, or sacbrood virus. Sixteen of 23 colonies (70.0%) were positive for black queen cell virus, and 21 (91.3%) had some evidence for deformed wing virus. No N. ceranae was detected in molecular screens of 55 colonies, although it is possible extremely low intensity infections exist; the more familiar N. apis was found in 53 colonies (96.4%). Under these conditions, K-wing was associated (positively) with colony strength; however, viruses and N. apis were not. Furthermore, black queen cell virus was positively and negatively associated with K-wing and deformed wing virus, respectively. Newfoundland honey bee colonies are thus free of several invasive parasites that plague operations in other parts of the world, and they provide a unique research arena to study independent pathology of the parasites that are present.

Infra-Population and -Community Dynamics of the Parasites Nosema apis and Nosema ceranae, and Consequences for Honey Bee (Apis mellifera) Hosts

Nosema spp. fungal gut parasites are among myriad possible explanations for contemporary increased mortality of western honey bees (Apis mellifera, hereafter honey bee) in many regions of the world. Invasive Nosema ceranae is particularly worrisome because some evidence suggests it has greater virulence than its congener N. apis. N. ceranae appears to have recently switched hosts from Asian honey bees (Apis cerana) and now has a nearly global distribution in honey bees, apparently displacing N. apis. We examined parasite reproduction and effects of N. apis, N. ceranae, and mixed Nosema infections on honey bee hosts in laboratory experiments. Both infection intensity and honey bee mortality were significantly greater for N. ceranae than for N. apis or mixed infections; mixed infection resulted in mortality similar to N. apis parasitism and reduced spore intensity, possibly due to inter-specific competition. This is the first long-term laboratory study to demonstrate lethal consequences of N. apis and N. ceranae and mixed Nosema parasitism in honey bees, and suggests that differences in reproduction and intra-host competition may explain apparent heterogeneous exclusion of the historic parasite by the invasive species