FINE-STRUCTURE AND MORPHOGENESIS OF THE MICROPYLE APPARATUS IN BEES EGGS

The present paper deals with the description of the formation of the micropylar apparatus in some species of Apidae bees. The features of the cells located in the anterior pole of the oocyte chamber are described at light microscopy and with SEM and TEM. The resulting micropylar region has the form of a sieved plate, slightly elevated in relationship to the oocyte surface. It is not clear if all the holes in the sieve are opened.

Ultrastructure and histochemistry of the mineral concretions in the midgut of bees (Hymenoptera: Apidae)

Mineral concretions in the digestive cells of bees were examined under transmission electron microscope and histochemically. Ultrastructure shows two types of mineral deposits: 1) mineral concretions which are organized in granules with a striking concentrically layered organization of opaque and clear zones and 2) electron dense granules which appear inside small vacuoles (0.4-0.7 mu m). These two structures are present in the apex of the digestive cells of the posterior midgut. Histochemical data reveal that mineral concretions are composed of calcium, iron and uric acid or its salts while calcium determination gives a positive reaction for electron dense granules. Morphological and chemical similarities between the mineral concretions of bees and those described for other insects suggest that they have an important physiological role regulating the composition of the internal environment and to avoid intoxication. Since concretions and granules are structurally distinct, it is suggested that they are functionally different.

A comparative study of digestive cells in different midgut regions of stingless bees (Hymenoptera: Apidae: Meliponinae)

Ultrastructure of the digestive cells was analyzed in three midgut regions (anterior, middle and posterior) of stingless bees. Variations occurs in the presence of lipid inclusions in the cells from posterior midgut and presence of double-membraned vesicles associated to microvilli in the anterior midgut. However, basal plasmic membrane infoldings and augmentation of surface area achieved by microvilli are very similar in all midgut regions. These results not supported the existence of fluid fluxes in the ectoperitrophic space and suggest that digestive cells in stingless bees are polifunctional, that is, there is not midgut region specialized in secretion or absorption as observed in other insects.

Heritability of sting characters in Africanized honeybees

In this paper we proposed to estimate the heritability of seven morphological characters that compose the sting apparatus ofthe Africanized honeybee workers. An experimental design to estimate genetic parameters was based on the method developed by Oldroyd and Moran. This method was modified to eliminate within-colony environmental effects associated with the additive genetic variance. The estimated h2 values ranged from 0.17 ± 0.11 (maximum width of bulb of sting stylet and height of the valve of right lancet) to 0.74 ± 0.30 (length of the lancet).

Cell death in ovarioles causes permanent sterility in Frieseomelitta varia worker bees

Frieseomelitta varia worker bees do not lay eggs even when living in queenless colonies, a condition that favors ovary development and oviposition in the majority of highly social bees. The permanent sterility of these worker bees was initially attributed to a failure in ovary morphogenesis and differentiation. Using transmission electron microscopy we found that at the beginning of the pupal phase the ovaries of F. varia workers are formed by four ovarioles, each of them composed of 1) a terminal filament at the apex of the ovarioles, containing juxtaposed and irregularly shaped cells, 2) a germarium with clusters of cystocytes and prefollicular cells showing long cytoplasmic projections that envelop the cystocyte clusters, 3) fusiform interfollicular and basal stalk precursor cells, and 4) globular, irregularly contoured basal cells with large nuclei. However, during the pupal phase an accentuated and progressive process of cell death takes place in the ovarioles. The dying cells are characterized by large membrane bodies, electron-dense apoptotic bodies, vacuoles, vesiculation, secondary lysosomes, enlarged rough endoplasmic reticulum cisternae, swollen mitochondria, pycnotic nuclei, masses of chromatin adjacent to the convoluted nuclear envelope, and nucleoli showing signs of fragmentation. Cell death continues in ovarioles even after the emergence of the workers. Once they become nurse bees, the ovaries have become transformed into a cell mass in which structurally organized ovarioles can no longer be identified. In F. varia workers, ovariole cell death most certainly is part of the program of caste differentiation.

Reciprocal interference between ants and stingless bees attending the honeydew-producing homopteran Aetalion reticulatum (Homoptera: Aetalionidae)

We evaluated the reciprocal effects between foragers of the ants Camponotus crassus and of the stingless bees Trigona hyalinata on aggregations of the honeydew-producing treehopper Aetalion reticulatum. The interactions were observed in Bauhinia variegata (Caesalpiniaceae) and Mangifera indica (Anacardiaceae) trees. We recorded the presence/absence of each attendant species in homopteran aggregations to test if the observed co-occurrence is lower than that expected by chance. An exclusion experiment was performed in which each attendant species was excluded from aggregations in order to test if an attendant species is more likely to occupy aggregations where the other attendant is not present. We also recorded the number of individuals of each attendant species in homopteran aggregations to search for any correlation between homopteran and attendant abundances. Additionally, we performed experiments using termites (Termitidae, Isoptera) as models to verify if the attendant species have the potential to defend A. reticulatum against natural enemies. The co-occurrence of attendant species was lower than that expected by chance. Homopteran aggregations without stingless bees were more visited by ants than those in which T. hyalinata was present, and vice-versa. The abundance of stingless bees was proportional to homopteran abundance, while ants abundance was not correlated to homopteran abundance. Both attendant species attacked the natural enemies models when we glued the termites ca. 1 cm away from homopteran aggregations, but only ants removed termites glued 5-7 cm away from aggregations. We suggest that the effects of non-formicid attendants should be included as another factor influencing the costs and benefits of ant-homopteran interactions, since honeydew availability for ants also depends on the presence and behavior of interspecific attendants.

Eficiência de extratos de plantas medicinais e aromáticas em atrair a abelha africanizada (Apis mellifera L.) em cultura de abacateiro (Persea americana Mili.)

The objective of the experiment was to test attractive substances for Africanized honey bees, Apis mellifera L., in avocado, Persea americana Mill. Were pulverized extracts of lemon grass (Cympobogon citratus), ocimum (Ocimum sellowii), Lippia alba, orange leaves (Citrus sinensis), eucaliptus leaves (Eucalyptus sp), eugenol and linalol and the frequency of bees on flowers were evaluated. The frequency of honey bees did increase with the application of the extracts falsa melissa, eucaliptus leaves and orange leaves, used as attractants in Quintal variety. The products tested in Eppendorf tubes did not increased the frequency the bees in Quintal and Fortuna varieties.

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.

Enzymatic variability among venoms from different subspecies of Apis mellifera (Hymenoptera: Apidae)

The enzymatic variability was analyzed in venom extracts from bees reared in different colonies of the Africanized, A. m. ligustica and A. m. carnica subspecies. The implications of this variation focused on the biochemistry differentiation and immunogenicity of these venoms. The results showed the existence of a huge variability among the subspecies as well as among the colonies for three out of the six tested components - hyaluronidase, acid phosphatase and proteases - suggesting the utilization of these features as possible biochemical markers. Furthermore, although not statistically significant, it was found that the Africanized bee venom presented slightly higher levels of protein content and esterase activity, when compared to the other subspecies. If the esterase plays a role in the pain intensity caused by the sting, as suggested elsewhere, this might suggest a reason for a bigger algogenicity of this venom in relation to that of European bees. On the other hand, A. m. ligustica bees presented the highest levels of proteolytic and acid phosphatase activities, whose functions are not enlightened in Hymenoptera venoms. The A. m. carnica workers presented the highest hyaluronidase and the lowest acid phosphatase activity levels. The extremely variable results among colonies of the same subspecies and among subspecies, for the tested venom components, justify the absence of correlation between allergic reactions and tests with pooled venom.

Hydrolases in the hypopharyngeal glands of workers of Scaptotrigona postica and Apis mellifera (Hymenoptera, Apinae)

Hydrolytic enzymes from hypopharyngeal gland extracts of newly emerged, nurse and foraging workers of two eusocial bees, Scaptotrigona postica, a native Brazilian stingless bee, and the Africanized honey bee (Apis mellifera) in Brazil, were compared. The hypopharyngeal gland is rich in enzymes in both species. Fifteen different enzymes were found in the extracts, with only a few quantitative differences between the species. Some of the enzymes present in the extracts may have intracellular functions, while others seem to be digestive enzymes. Scaptotrigona postica, had lower β-glucosidase and higher lipase esterase activities than A. mellifera. The differences may be due to different feeding habits and behavioral peculiarities of the two species. ©FUNPEC-RP.

Éster de sacarose no controle do Varroa estructor em abelhas africanizadas

This study aimed to determine the effect of sucrose ester on the control of Varroa destructor mite infestation in Africanized honeybees. For the in vitro experiments, the product was tested in bees and mites at five concentrations obtained through dilution in water (T0: 100% distilled water; T1: 0.5%; T2: 1%; T3: 2%; T4: 5%; and T5: 10% sucrose ester).For the field studies, the experimental design was completely randomized, with four treatments and seven replicates, totaling 28 colonies, from which seven were the controls, seven were treated with 0.1% sucrose ester, seven with 0.2% sucrose ester, and seven hives with 0.5% sucrose ester diluted in water. In the in vitro study, the sucrose ester at 0.5% concentration caused mite and bee mortality. In the field tests, the product at 0.2% concentration reduced Varroa destructor infestation in Africanized honeybees and, therefore, may be used as a tool to control this pest. At 0.1, 0.2, and 0.5% concentrations, sucrose ester did not impair the establishment of open and capped brood areas, as well as stored food areas in the hive, suggesting it is not toxic to Africanized honeybees.

Anticancer effects of geopropolis produced by stingless bees on canine osteosarcoma cells in vitro

Geopropolis is produced by indigenous stingless bees from the resinous material of plants, adding soil or clay. Its biological properties have not been investigated, such as propolis, and herein its cytotoxic action on canine osteosarcoma (OSA) cells was evaluated. OSA is a primary bone neoplasm diagnosed in dogs being an excellent model in vivo to study human OSA. spOS-2 primary cultures were isolated from the tumor of a dog with osteosarcoma and incubated with geopropolis, 70% ethanol (geopropolis solvent), and carboplatin after 6, 24, 48, and 72 hours. Cell viability was analyzed by the crystal violet method. Geopropolis was efficient against canine OSA cells in a dose- and time-dependent way, leading to a distinct morphology compared to control. Geopropolis cytotoxic action was exclusively due to its constituents since 70% ethanol (its solvent) had no effect on cell viability. Carboplatin had no effect on OSA cells. Geopropolis exerted a cytotoxic effect on canine osteosarcoma, and its introduction as a possible therapeutic agent in vivo could be investigated, providing a new contribution to OSA treatment. © 2013 Naiara Costa Cinegaglia et al.

Proteome and phosphoproteome of Africanized and European honeybee venoms

Honey bee venom toxins trigger immunological, physiological, and neurological responses within victims. The high occurrence of bee attacks involving potentially fatal toxic and allergic reactions in humans and the prospect of developing novel pharmaceuticals make honey bee venom an attractive target for proteomic studies. Using label-free quantification, we compared the proteome and phosphoproteome of the venom of Africanized honeybees with that of two European subspecies, namely Apis mellifera ligustica and A. m. carnica. From the total of 51 proteins, 42 were common to all three subspecies. Remarkably, the toxins melittin and icarapin were phosphorylated. In all venoms, icarapin was phosphorylated at the 205Ser residue, which is located in close proximity to its known antigenic site. Melittin, the major toxin of honeybee venoms, was phosphorylated in all venoms at the 10Thr and 18Ser residues. 18Ser phosphorylated melittin-the major of its two phosphorylated forms-was less toxic compared to the native peptide. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Porcentagem de aceitação de larvas de diferentes grupos genéticos de Apis mellifera na produção de abelhas rainhas

Pós-graduação em Zootecnia - FMVZ