Produtos naturais para o controle do ácaro Varroa destructor em abelhas africanizadas

O objetivo deste trabalho foi avaliar os efeitos do ácido oxálico e de óleos essenciais de plantas no controle da infestação pelo ácaro Varroa destructor em colônias de Apis mellifera africanizadas. O experimento foi realizado em delineamento inteiramente casualizado, em 30 colônias, com seis tratamentos e cinco repetições. As colmeias foram tratadas com óleos essenciais de arruda (Ruta graveolens), eucalipto (Eucalyptus spp.) e hortelã (Mentha piperita), além de timol, ácido oxálico e do tratamento controle sem aplicação de produtos. Foram avaliadas a mortalidade de varroas e as taxas de mortalidade de crias e de infestação de varroas em crias e em abelhas adultas, antes e depois da aplicação de cada produto. O ácido oxálico e os óleos de arruda, timol, eucalipto e de hortelã reduziram a mortalidade de crias parasitadas pelo ácaro em 92,1, 83,3, 81,7, 86,4 e 81,3%, respectivamente. O tratamento com ácido oxálico reduziu em 87,4% a infestação de varroas em abelhas adultas. O uso desses produtos é eficiente na redução da mortalidade de crias de A. mellifera parasitadas por V. destructor.

Atividade antimicrobiana de própolis de Apis mellifera obtidas em três regiões do Brasil

As propriedades biológicas da própolis de Apis mellifera são amplamente relatadas sendo comuns variações nas mesmas em função da região onde foram produzidas. A ação antimicrobiana de própolis obtidas em três regiões do Brasil (Botucatu-SP, Mossoró-RN e Urubici-SC) foi investigada sobre linhagens isoladas de infecções clínicas humanas (Staphylococcus aureus, Escherichia coli, Enterococcus sp, Pseudomonas aeruginosa e Candida albicans). Foram preparados extratos alcoólicos de própolis (EAP) e determinada a Concentração Inibitória Mínima (CIM) seguida do cálculo da CIM90%. A própolis de Botucatu foi a mais eficiente sobre S. aureus (0,3%v/v), Enterococcus sp (1,1%v/v) e C. albicans (2,1% v/v). Para E. coli, a própolis eficiente foi de Urubici (7,0%v/v) e para P. aeruginosa a de Mossoró (5,3%v/v). Os resultados mostram maior sensibilidade das bactérias Gram positivas e levedura em relação às Gram negativas. É possível concluir que, para os microrganismos testados e amostras de própolis testadas, há diferenças na atividade antimicrobiana em função do local de produção e que isso se explica pela diferença de composição química da própolis.

Cytotoxic constituents of propolis inducing anticancer effects: a review

Objectives Propolis is a honeybee product used extensively in traditional medicine for its antioxidant, anti-inflammatory, immunomodulatory and anticancer effects. Propolis exhibits a broad spectrum of biological activities because it is a complex mixture of natural substances. In this review, the antitumour effects of propolis extracts and its constituents (e. g. flavonoids, terpenes and caffeic acid phenethyl ester) are discussed.Key findings The effect of propolis on experimental carcinogenesis is discussed, as well as its possible mechanisms of action against tumours, involving apoptosis, cell cycle arrest and interference on metabolic pathways. Propolis seems to be efficient against different tumour cells both in vitro and in vivo, which suggests its potential in the development of new anticancer drugs.Summary Propolis extracts may be important economically and would allow a relatively inexpensive cancer treatment. Preclinical investigations are needed to further elucidate the benefits of propolis and its antitumour properties.

The effects of propolis on antibody production by laying hens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Insect Venom Peptides

The insects of the order Hymenoptera ( bees, wasps, and ants) are classified in two groups, based on their life history: social and solitary. The venoms of the social Hymenoptera evolved to be used as defensive tools to protect the colonies of these insects from the attacks of predators. Generally they do not cause lethal effects but cause mainly inflammatory and/or immunological reactions in the victims of their stings. However, sometimes it is also possible to observe the occurrence of systemic effects like respiratory and/or kidney failure. Meanwhile, the venoms of solitary Hymenoptera evolved mainly to cause paralysis of the preys in order to permit egg laying on/within the prey's body; thus, some components of these venoms cause permanent/transient paralysis in the preys, while other components seem to act preventing infections of the food and future progenies. The peptide components of venoms from Hymenoptera are spread over the molar mass range of 1400 to 7000 da and together comprise up to 70% of the weight of freeze-dried venoms. Most of these toxins are linear polycationic amphipatic peptides with a high content of alpha-helices in their secondary structures. These peptides generally account for cell lysis, hemolysis, antibiosis, and sometimes promote the delivery of cellular activators/mediators through interaction with the G-protein receptor, and perhaps some of them are even immunogenic components. In addition to these peptides, the Hymenopteran venoms also may contain a few neurotoxins that target Na+ and/or Ca+2 channels or even the nicotinic ACh receptor. This review summarizes current knowledge of the biologically active Hymenoptera venoms.

Morphometric analysis of the venom gland in worker bees of Apis mellifera (Hymenoptera : Apidae) from the pantanal region of Mato Grosso do Sul

In Apis mellifera the acid or venom gland is composed of secretory cells that surround a channel that opens into a reservoir devoid of musculature. This gland can at times present apical branching. In this study we recorded the frequency of branched venom glands in workers of Africanized bees (Apis mellifera Linnaeus) from six localities in the Pantanal region of Mato Grosso do Sul, and analyzed the relation among the length of the main duct, the length of the duct from the reservoir to the beginning of branching, the length of the branched segment (when present) and the total length of the gland. We sought to determine the probable genotypes of the bees from each population by using the model proposed by Alves-Junior. The frequency of branched glands varied from 50% to 83% in the worker bees coming from those places, indicating that this characteristic is primitive in these bees. The results of the Analysis of Discriminant Functions indicated significant differences in the morphometrical segments of the venom gland (Wilk's Lambda = 0.065; F-(27,F-30) = 4.507; P < 0.001), and permitted a differentiation of the populations studied. The genotypes inferred for the bees of each locality agree with the results obtained in the Analysis of Discriminant Functions and form three distinct groups, with some overlapping areas among them. In all of the populations considered the phenotype largevenom gland was predominant. It is inferred that bees with this phenotype (venom gland larger than S. 15 mm) have Gm(1) Gm(1) genotype, being therefore homozygotes for the major alleles and also for the modifier genes that codify this morphological trait. The high frequency of worker bees with large venom gland in all the places considered makes viable the development of a selection program in order to obtain bees with longer venom glands, aimed at the commercial production of venom by the beekeepers of the Pantanal region of Mato Grosso do Sul.

Effect of narcosis with CO2 on the ovarian development in queens of Apis mellifera (Hymenoptera, Apini)

Exposure of 1, 4, 7 & 10 day-old virgin queens of Apis mellifera Linne 1758 for 1 min to CO2 accelerated their ovarian development, having a similar effect as mating on the initial formation of the ovarian follicles. In 3 day-old queens the exposure to CO2 enhanced the initial stage of germ cell differentiation into oocytes and nurse cells in the ovarioles, a developmental stage only seen in 5 day-old untreated virgin queens, the age at which they are ready to mate. In 10 day-old untreated virgin queens, some regions of the ovarioles presented tissue disorganization and many cells with pycnotic nuclei. However, narcotized virgin queens of the same age did not present such ovary degeneration. Conversely, they showed nitid follicle formation, arising in the ovarioles' initial differentiation between nurse and oocytic chambers, although still without vitellogenesis. The accelerative effect of CO2 is limited to the ages near to those proper for mating, since 15 and 18 day-old treated virgin queens presented ovaries with extensive regions of high tissue disorganization and a great number of cells with pycnotic nuclei. According to the results, the narcosis presented three levels of effect on the ovary of honeybee virgin queens: 1) accelerated the germ cell differentiation, 2) preserved the tissue integrity even after the queen mating period and 3) stimulated the initial differentiation of a vitellarium. This later condition was only seen in untreated queens after mating. All these effects are not maintained if the queen is kept virgin beyond 15 days old.

Differences in the number of some antennal sensilla of four honey bee (Apis mellifera) types and comparisons with the defensive behaviour

The number of sensilla coeloconica, ampullacea and campaniformia of flagellomeres 3 to 10 of the antennae of workers of four honey bee types (Italian, Caucasian, African and Africanized) was studied by scanning electron microscopy. Comparisons of the four bee types showed that only African and Africanized honey bees did not differ from one another with respect to the number of sensilla coeloconica and ampullacea of flagellomere 10. African and Africanized honey bees and Caucasian and Italian honey bees also did not differ from one another in terms of flagellomere 9. In the other flagellomeres there were no differences among bee types. Italian and Caucasian honey bees differed from Africanized honey bees in terms of number of sensilla campaniformia on flagellomere 6, and Caucasian honey bees differed from African and Africanized honey bees in terms of flagellomere 3. Five significant but random correlation values were obtained between number of antennal sensilla and defensive behaviour in Africanized honey bees. Thus,there is no relationship between antennal structures and defence behaviour.

Enzymatic activity of hypopharyngeal gland extractsfrom workers of Apis mellifera (Hymenoptera, Apidae, Apinae)

This research presents a comparative study of enzymatic activity of the hypopharyngeal gland extracts from workers of Apis mellifera in three physiologic stages: newly emerged, nurse and forager workers, with the objective of contributing to the comprehension of the gland function. In order to determinate the enzymes present in the extracts, the Api Zym kit (Bio Merieux) was used to test the activity of 19 different enzymes. The enzymes found in larger amounts only in the hypopharyngeal glands from certain individuals were the following: in newly emerged workers, the N-acetyl-double down arrow-glucosaminidase that may be digesting the chitin of some food ingested by the bee; in forager workers, the acid phosphatase that is likely acting in authophagic processes, the a-glucosidase, in the processing of nectar into honey, and the double down arrow-glucosidases, in the pollen digestion.

Alterations induced by the juvenile hormone in glandular cells of the Apis mellifera venom gland: Applications on newly emerged workers (Hymenoptera, Apidae)

Histological and histochemical analyses were carried out in order to evaluate the influence of the topical application of a synthetic juvenile hormone on the secretory cycle and degeneration of the venom gland of Apis mellifera. Newly emerged workers received the topical application of synthetic hormone and the results were compared to the normal development of the secretory cycle in virgin and mated queens. The first worker group received the juvenile hormone diluted in hexane (2 mu g/mu L), the second received only mu L of hexane, and the third did not receive any kind of application. After the application the workers were returned to the colony and collected at the ages of 14 and 25 days of adult life. The groups with virgin queens and the other with mated queens, did not receive the treatment. The results show that the individuals treated with juvenile hormone and with pure hexane presented differences in the histological and cytochemical aspects of the secretory cells of the venom gland. The data indicate that both the juvenile hormone and hexane accelerate the activity of the secretory cycle and the degeneration of the venom gland; however, the juvenile hormone proved to be more effective than hexane. (c) 2006 Elsevier Ltd. All rights reserved.

Morphological characterization of cell death during the ovary differentiation in worker honey bee

Cell death that occurs during ovary differentiation in the honeybee worker's larval development accounts for ovariole reabsorption. From a morphological standpoint, three modes of death were detected. Germinative cells in the ovarioles die by an apoptotic-like process, whereas the somatic cells die by an autophagic process, type 11 cell death; and during pupation, stromatic and ovarian capsular cells die through cytoplasmic disintegration, releasing their components into the hemolymph. These modes of cell death are in part determined by the pattern of tissue organization within which the cell occurs. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Jelleines: a family of antimicrobial peptides from the Royal Jelly of honeybees (Apis mellifera)

Four antimicrobial peptides were purified from Royal Jelly of honeybees, by using reverse phase-HPLC and sequenced by using Q-Tof-MS/MS: PFKLSLHL-NH2 (Jelleine-I), TPFKLSLHL-NH2 (Jelleine-II), EPFKLSLHL-NH2 (Jelleine-III), and TPFKLSLH-NH2 (Jelleine-IV). The peptides were synthesized on-solid phase, purified and submitted to different biological assays: antimicrobial activity, mast cell degranulating activity and hemolysis. The Jelleines-I-III presented exclusively antimicrobial activities against yeast, Gram+ and Gram- bacteria; meanwhile, Jelleine-IV was not active in none of the assays performed. These peptides do not present any similarity with the other antimicrobial peptides from the honeybees; they are produced constitutively by the workers and secreted into Royal Jelly. (C) 2004 Elsevier B.V. All rights reserved.

Structure determination of hydroxytrypargine: A new tetrahydro-beta-carboline toxin from the venom of the spider Parawixia bistriata

A new, highly active tetrahydro-p-carboline toxin from the spider Parawixia bistriata, the most-common species of social spider occurring in Brazil, was isolated. The new toxin was identified as 1,2,3,4-tetrahydro-6-hydroxy-beta-carboline (= N-[3-(2,3,4,9-tetrahydro-6-hydroxy-1H-pyrido[3,4-b]indol-1-yl)propyl]guanidine; 3). This type of alkaloid, not common among spider toxins, was found to be the most-potent constituent of the spider's chemical weaponry to kill prey. When P bistriata catch arthropods in their web, they apparently attack their prey in groups of many individuals injecting their venoms. In vivo toxicity assays with 3 demonstrated a potent lethal effect to honeybees, giving rise to clear neurotoxic effects (paralysis) before death. The compound's toxicity (LD50 value) was determined to be ca. 8 ng/g of honeybee. The investigation of the pharmacological properties and neurotoxic actions of 3 may be used in the future for the development of new drugs to be applied for pest control in agriculture.

The venomous secrets of the web droplets from the viscid spiral of the orb-weaver spider Nephila clavipes (Araneae, Tetragnatidae)

The capture web of N. clavipes presents viscous droplets, which play important roles in web mechanics and prey capture. By using scanning and transmission electron microscopy, it was demonstrated that the web droplets are constituted of different chemical environments, provided by the existence both of an aqueous and a lipid layer, which, in turn, present a suspension of tenths of vesicles containing polypeptides and/or tipids. GC/EI-MS Analysis of the contents of these vesicles led to the identification of some saturated fatty acids, such as decanoic acid, undecanoic acid, dodecanoic acid, tetradecanoic acid, octadecanoic acid, and icosanoic acid, while other components were unsaturated fatty acids, such as (Z)-tetradec-9-enoic acid, (Z)-octadec-9-enoic acid, and (Z)-icosa-11-enoic acid; and polyunsaturated fatty acids like (9Z,12Z)-octadeca-9,12-dienoic acid, (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid, and (11Z,14Z)-icosa-11,14-dienoic acid. Toxic proteins such as calcium-activated proteinase and metalloproteinase jararhagin-like precursor were also identified by using a proteomic approach, indicating the possible involvement of these enzymes in the pre-digestion of spiders' preys web-captured. Apparently, the mixture of fatty acids are relatively toxic to insects by topical application (LD50 64.3 +/- 7.6 ng mg(-1) honeybee), while the proteins alone present no topical effect; however, when injected into the prey-insects, these proteins presented a moderate toxicity (LD50 40.3 +/- 4.8 ng mg(-1) honeybee); the mixture of fatty acids and proteins is very toxic to the preys captured by the web droplets of the viscid spiral of Nephila clavipes when topically applied on them (LD50 14.3 +/- 1.8ng mg(-1) honeybee).

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.