Differences in the number of sensilla coeloconica and sensilla ampullacea of segment 10 of the antennae of Africanized and Caucasian bees and of their F1 hybrids

The numbers of sensilla coeloconica and sensilla ampullacea of segment 10 of the antennae of Caucasian and Africanized worker bees and of their hybrids were counted with the aid of a scanning electron microscope. Africanized bees have fewer sensilla than Caucasian bees and the continuous distribution obtained from the F1 values suggests polygenic inheritance for the control of this trait. There are also indications that the hybrid Brazilian Apis mellifera studied here are close to typical African bees (Apis mellifera scutellata) in terms of number of sensilla.

Polybitoxins: A group of phospholipases A2 from the venom of the neotropical social wasp paulistinha (Polybia paulista)

The neotropical wasp Polybia paulista is very aggressive and endemic in south-east Brazil, where it frequently causes stinging accidents. By using gel filtration on Sephadex G-200, followed by ion-exchange chromatography on DEAE-Cellulose under a pH gradient, a group of four toxins (designated as polybitoxins-I, II, lII and IV) presenting phospholipase A2 (PLA2) activities was purified. These toxins are dimeric with mol. wts ranging from 115,000 to 132,000 and formed by different subunits. The four toxins contain very high sugar contents attached to their molecules (22-43% w/w) and presented different values of pH optimum from 7.8 to 9.0; when dissociated, only residual catalytic activities were maintained. The catalytic activities of polybitoxins (from 18 to 771 μmoles/mg per minute) are lower than that of PLA2 from Apis mellifera venom and hornetin from Vespa basalis. The polybitoxins presented a non-linear steady-state kinetic behavior for the hydrolysis of phosphatidylcholine at pH 7.9, compatible with the negative co- operativity phenomena. All of the polybitoxins were very potent direct hemolysins, especially the polybitoxins-III and IV, which are as potent as the lethal toxin from V. basalis and hornetin from Vespa flavitarsus, respectively; polybitoxin-IV presented hemolytic action 20 times higher than that of PLA2 from A. mellifera, 17 times higher than that of neutral PLA2 from Naja nigricolis and about 37 times higher than that of cardiotoxin from Naja naja atra venom.

Differential morphology of the hemocytes present in pupal ovaries of the queen and worker honeybee

The location and morphological features of the blood cells found in the pupal ovary of workers and queens of Apis mellifera are described in relationship with their probable function in the ovary differentiation. The hemocytes from inside the ovarioles are different in both castes. In queens their morphology suggest an action in the tunica propria production, while in workers it suggest a phagocytic activity. The hemocytes present in the intersticial tissue are phagocytes in both castes, and may be responsible by the ovary shapping during metamorphosis.

Enzymes in the hypopharyngeal gland extracts from workers of Scaptotrigona postica (Hymenoptera, Apinae, Meliponini) related to food storing in the colony

This study reports on research of enzymes produced by the hypopharyngeal glands, which are related to food storing in the colony, from gland extracts from nurse and forager workers of S. postica. Only the presence of the saccharase was detected in the extracts from the glands of forager workers. The results were compared to the enzymatic content of similar extracts of A. mellifera taking into account the behavioral differences among the two species.

Contribuição ao protocolo de controle de qualidade da própolis e de seus extratos

A quality control protocol for the analysis of propolis and its extracts is proposed in the present work. Propolis of Apis mellifera L. bees collected in beehives of an apiary located in the northwest of the Paraná State was triturated and submitted to the following analysis: particles medium size determination, loss of dry, ashes drift, waxes drift, drift of extractive (in water and in ethanol) and total flavonoids drift determination. Propolis ethanolic extracts (96 °GL) at 10% (w/w) and at 30% (w/w) were prepared and submitted to the determination of the pH, relative density, dry residue, alcoholic drift and total flavonoids drift determination. Propolis was analyzed, through High Performance Liquid Cromatography (HPLC). Comparing the obtained results with other works, it was observed that is possible to establish the intervals of values for parameters in order to evaluate the quality of a propolis sample and its extracts.

Enzymatic activity of the hypopharyngeal gland extract from workers and males of Scaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponini)

The objective of this research was to contribute to elucidation of the function of the hypopharyngeal glands of S. postica in enzyme production, using the Api Zym kit (Bio Mérieux). Dealing with a comparative analysis between the enzymatic content of the hypopharyngeal gland extracts from newly emerged, nurse and forager workers, as well as, newly emerged males and males mature for mating of S. postica. The hypopharyngeal glands from nurse workers of Apis mellifera, that produce part of the royal jelly, were used for comparison. While in A. mellifera, the hypopharyngeal glands are present only in workers, in S. postica, the hypopharyngeal glands are present and functioning in all adult individuals of the colony. The higher enzymatic activity was observed in the hypopharyngeal gland extracts from nurse workers and may be related to a larger demand for energy, compared to other individuals. The occurrence of large quantities of leucine arylamidase in all individuals may mean that protein processing is happening.

Is the number of antennal plate organs (sensilla placodea) greater in hygienic than in non-hygienic Africanized honey bees?

Hygienic behavior is a desirable trait in honey bees (Apis mellifera L.), as hygienic bees quickly remove diseased brood, intermpting the infectious cycle. Hygienic lines of honey bees appear to be more sensitive to the odors of dead and diseased honey bee brood, and Africanized honey bees are generally more hygienic than are European honey bees. We compared the number of sensilla placodea, antennal sensory structures involved in the perception of odor, in 10 bees from each of six hygienic and four non-hygienic colonies of Africanized honey bees. The sensilla placodea of three of the terminal segments (flagellomeres) of the right antenna of each bee were counted with a scanning electron microscope. There were no significant differences in the mean numbers of sensilla placodea between the hygienic and non-hygienic bees, though the variance was higher in the hygienic group. Flagellomere 4 had significantly more sensilla placodea than flagellomeres 6 and 8. However, there was no significant difference between the other two flagellomeres. As hygienic bees are capable of identifying dead, injured, or infested brood inside a capped brood cell, sensilla placodea probably have an important role in enabling worker bees to sense sick brood. However, we did not find greater numbers of this sensory structure in the antennae of hygienic, compared to non-hygienic Africanized honey bees.

Development of worker ovaries in Brazilian Africanized honeybees (Hymenoptera: Apidae)

The present paper compares the worker ovaries development in queen right and orphaned colonies of Brazilian Africanized honeybee. The results show that in both colonies some workers starting 20 days after emergence, present some kind of ovary development. As the workers from queen right colony get older the degree of the ovary development increases. Vitellogenesis in worker ovaries may occur in queen right and orphaned colonies, but its frequency is lower in queen right colonies.

Morphology and protein patterns of honey bee drone accessory glands.

We used light and transmission electron microscopy to examine the morphology of the accessory glands of immature and mature adult males of Apis mellifera L. We also made an electrophoretic analysis of the protein content of the mature gland. The glands of the immature male actively secrete a mucous substance that can be seen in the lumen of the gland of the mature male. This secretion stains with mercury bromophenol blue and with periodic acid-Schiff reaction, which stain glyconjugates. The protein content was higher in the lumen secretion than in the gland wall extracts. The electrophoresis patterns of the wall extracts were different from those of the secretion found in the gland lumen.

Africanized honeybees: Biological characteristics, urban nesting behavior and accidents caused in Brazilian cities (Hymenoptera: Apidae)

In 1956 African honeybee queens (Apis mellifera scutellata) were imported from South Africa and Tanzania to Brazil, as part of a government project to increase Brazilian honey production. The European honeybees existing in that country had not adapted well to the tropical conditions and consequently, had a low productivity. The newly introduced bee was known to produce substantially more honey than the other subspecies, but was also famous for its great aggressiveness and quicker attack of intruders with less disturbance. Hoping to create a new hybrid bee that would be both docile and productive, the scientist Warwick Estevam Kerr tried to cross the African and the European subspecies under controlled conditions. However, an accident resulted in the escape of 26 swarms into the Brazilian countryside, where their queens mated with drones of the European resident honeybees. The poly-hybrid bees resulting from these crossings expressed scutellata-like reproductive, foraging, and defensive behaviors and, for this reason, were called Africanized honeybees. They spread rapidly from the introduction area of the African honeybees (near Rio Claro, São Paulo state) to as far south as mid-Argentina and to the north of Texas, also settling in Arizona, New Mexico, California and Nevada, due to their high adaptability to variable ecological conditions. In spite of a few undesirable behaviors, these bees have been invoking larger economic interest because they produce much more honey, have good resistance to diseases and are excellent pollinators. In Brazil, because people frequently disturb the environment, the occupation of urban refuges by Africanized honeybees has been increasing in the last years. The concern with accidents is generally associated with the high swarming frequency recorded during the year and the variety of shelters available in urban areas. This paper deals with the biological characteristics of the Africanized honeybees, their nesting behavior in urban environments, and accidents caused by these bees in Brazilian cities.

Composition of stingless bee honey: Setting quality standards

Compositional data from 152 stingless bee (Meliponini) honey samples were compiled from studies since 1964, and evaluated to propose a quality standard for this product. Since stingless bee honey has a different composition than Apis mellifera honey, some physicochemical parameters are presented according to stingless bee species. The entomological origin of the honey was known for 17 species of Meliponini from Brazil, one from Costa Rica, six from Mexico, 27 from Panama, one from Surinam, two from Trinidad & Tobago, and seven from Venezuela, most from the genus Melipona. The results varied as follows: moisture (19.9-41.9g/100g), pH (3.15-4.66), free acidity (5.9-109.0meq/Kg), ash (0.01-1.18g/100g), diastase activity (0.9-23.0DN), electrical conductivity (0.49-8.77mS/cm), HMF (0.4-78.4mg/Kg), invertase activity (19.8-90.1IU), nitrogen (14.34-144.00mg/100g), reducing sugars (58.0-75.7g/100g) and sucrose (1.1-4.8g/100g). Moisture content of stingless bee honey is generally higher than the 20% maximum established for A. mellifera honey. Guidelines for further contributions would help make the physicochemical database of meliponine honey more objective, in order to use such data to set quality standards. Pollen analysis should be directed towards the recognition of unifloral honeys produced by stingless bees, in order to obtain standard products from botanical species. A honey quality control campaign directed to both stingless beekeepers and stingless bee honey hunters is needed, as is harmonization of analytical methods. © 2007 Asociación Interciencia.

Propriedade físico-química da própolis em função da sazonalidade e método de produção

The aim of this work was to investigate monthly, for one year the seasonality effect on the physicochemical properties (dry weight, flavonoids contents, pH and antioxidant activity) of ethanolic extract (EAP) of propolis produced by three different techniques (intelligent propolis collector, plastic screen or scraping) fifteen colonies of africanized bees. No differences (p>0.05) in the physicochemical properties due to the technique of propolis production and seasonality were observed.

Hyaluronidase from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae): Cloning, structural modeling, purification, and immunological analysis

In this study, we describe the cDNA cloning, sequencing, and 3-D structure of the allergen hyaluronidase from Polybia paulista venom (Pp-Hyal). Using a proteomic approach, the native form of Pp-Hyal was purified to homogeneity and used to produce a Pp-specific polyclonal antibody. The results revealed that Pp-Hyal can be classified as a glycosyl hydrolase and that the full-length Pp-Hyal cDNA (1315 bp; GI: 302201582) is similar (80-90%) to hyaluronidase from the venoms of endemic Northern wasp species. The isolated mature protein is comprised of 338 amino acids, with a theoretical pI of 8.77 and a molecular mass of 39,648.8 Da versus a pI of 8.13 and 43,277.0 Da indicated by MS. The Pp-Hyal 3D-structural model revealed a central core (α/β)7 barrel, two sulfide bonds (Cys 19-308 and Cys 185-197), and three putative glycosylation sites (Asn79, Asn187, and Asn325), two of which are also found in the rVes v 2 protein. Based on the model, residues Ser299, Asp107, and Glu109 interact with the substrate and potential epitopes (five conformational and seven linear) located at surface-exposed regions of the structure. Purified native Pp-Hyal showed high similarity (97%) with hyaluronidase from Polistes annularis venom (Q9U6V9). Immunoblotting analysis confirmed the specificity of the Pp-Hyal-specific antibody as it recognized the Pp-Hyal protein in both the purified fraction and P. paulista crude venom. No reaction was observed with the venoms of Apis mellifera, Solenopsis invicta, Agelaia pallipes pallipes, and Polistes lanio lanio, with the exception of immune cross-reactivity with venoms of the genus Polybia (sericea and ignobilis). Our results demonstrate cross-reactivity only between wasp venoms from the genus Polybia. The absence of cross-reactivity between the venoms of wasps and bees observed here is important because it allows identification of the insect responsible for sensitization, or at least of the phylogenetically closest insect, in order to facilitate effective immunotherapy in allergic patients. © 2013 Elsevier Ltd.

Effects of sublethal dose of fipronil on neuron metabolic activity of africanized honeybees

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. © 2012 Springer Science+Business Media New York.

Age and Morphological Changes in the Epiponini Wasp Polybia paulista Von Ihering (Hymenoptera: Vespidae)

The division of labor is a central theme in the study of social insects. In bees and wasps, this activity is regulated by age polyethism. Important physiological and morphological changes have been widely studied in the polyethism of honeybee workers. In contrast, this is a relatively unexplored subject in social vespids. Our goal was to determine if there are detectable morphological changes in the body of the Epiponini wasp Polybia paulista Von Ihering or in certain glands in relation to age polyethism. We observed changes in the body weight, the salivary gland, and the mandibular gland that were associated with age, and our results suggest that social relationships and task performance are important to these changes. This contrasts with observations in Polistes and is different from the Apis mellifera Linnaeus age polyethism model. © 2013 Sociedade Entomológica do Brasil.