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.

Bioinsecticide action of the venom of Africanized bees Apis mellifera Linnaeus 1758 (Hymenoptera: Apidae): I - The most susceptible age of Diatraea saccharalis Fabricius 1794 (Lepeletier: Pyralidae) eggs to the venom action

The present paper aimed at testing the action of non-lyophilized venom of Africanized bees Apis mellifera through topical applications on Diatraea saccharalis egg masses. The CL50, DL50 and the most susceptible age of eggs to the venom topic application were also determined. Three-day-old eggs were the most susceptible to the venom action with CL50 equal to 8.6 mg/ml and DL50 equal to 0.173 mg/mass. The venom loses its action after being stored for 15 days.

Geographic variation of Africanized honey bees (Apis mellifera L.) in Brazil: Multivariate morphometrics and racial admixture

The correspondence between morphometric and isozymic geographic variation patterns of Africanized honey bees in Brazil was analyzed. Morphometric data consisted of mean vectors of 19 wing traits measured in 42 local populations distributed throughout the country. Isozymic data refer to allelic frequencies of malate dehydrogenase (MDH), and were obtained from Lobo and Krieger. The two data sets were analyzed through canonical trend surface, principal components and spatial autocorrelation analyses, and showed north-south dines, demonstrating that Africanized honey bees in southern and southeastern Brazil are more similar to European honey bees than those found in northern and northeastern regions. Also, the morphometric variation is within the limits established by the racial admixture model, considering the expected values of Africanized honey bee fore wing length (WL) in southern and northeastern regions of Brazil, estimated by combining average values of WL in the three main subspecies involved in the Africanization process (Apis mellifera scutellata, A. m. ligustica and A. m. mellifera) with racial admixture coefficients.

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.

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.

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.

Distribution of acid phosphatases in the hypopharyngeal glands from workers, queens and males of a Brazilian stingless bee Scaptotrigona postica Latreille: An ultrastructural cytochemical study

The present study reports the localization of acid phosphatase in the hypopharyngeal gland cells from workers (newly-emerged, nurse and forager), queens (newly-emerged and laying) and males (newly-emerged and mature for mating) of the Brazilian stingless bee, Scaptotrigona postica. The phosphatase activity varied in intensity and localization depending on the individual class, physiological age and the substrate used. In newly-emerged workers, the phosphatase-positive sites suggest the involvement of the enzyme with cellular differentiation that occurs in the presecretory phase, in nurse workers with protein synthesis and in forager workers with changes in cellular activity or glandular regression. In males mature for mating and laying queens, the positive sites are related to secretory activity, showing that the gland maintains some activity in spite of the regressive aspect. Of the substrates used, β-glycerophosphate gave the least specific localization.

Enzymes present in the thoracic gland extracts from workers and males of Apis mellifera (Hymenoptera: Apidae)

This research deals with the analysis of the enzymes present in thoracic gland extracts from newly emerged, nurse workers, forager workers, newly emerged males, and mature males of A. mellifera L. (Hymenoptera, Apoidea, Apidae). The enzymes found in larger quantities in the thoracic gland occurred in all classes of workers and are digestive. Acid phosphatase and Naphtol-AS-BI-phosphohydrolase act in protein synthesis, leucine arylamidase hydrolyses proteins and a-glucosidase actuate in the nectar processing into honey. Naphtol-AS-BI-phosphohydrolase was found in larger quantities only in workers, this suggests action in protein synthesis by the thoracic gland, b-galactosidase is in larger amounts in the newly emerged bees (workers and males) this aids in the provision of other substances to be used as an energy source when glucose or sucrose are absent. Differences between enzymatic profiles from workers and males are usually related to their colony tasks, or related to their physiological necessities per individual in specific life stages.

Enzymatic activity of hypopharyngeal gland extracts from 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 Mérieux) 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-down double arrow sign-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 down double arrow sign-glucosidases, in the pollen digestion.

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.

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.

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.

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.