A specialized midgut region in stingless bees (Hymenoptera: Apidae: Meliponinae)

Ultrastructure of the anterior end of the midgut of three Neotropical stingless bees, including an obligate necrophagous species Trigona hypogea, suggests that this midgut region has no role in nutrient absorption because its lumen is reduced by a thick cuticle that lines the outer epithelium of the cardiac valve. A possible involvement of this midgut region in the production of the peritrophic membrane compounds is discussed.

Evaluation of a larval development assay for the detection of anthelmintic resistance in Ostertagia circumcincta

Two experiments were carried out to evaluate a larval development assay for the detection of anthelmintic resistance in O. circumcincta. In Experiment I, the dose responses to levamisole (LEV), thiabendazole (TBZ) and ivermectin (IVM) of 8 isolates of O. circumcincta were measured 34 days after infection (DAI). Four of these isolates were shown to be resistant to 1 or more anthelmintics. With 2 exceptions, all isolates considered to be resistant had higher LD50 values than the susceptible isolates for that anthelmintic. One exception was isolate RM8, which was considered to be resistant to all 3 anthelmintics based on faecal egg count reduction tests in goats, but the LD50 value for LEV did not differ from that for the susceptible isolates. The other exception was an isolate considered to be susceptible to TBZ which had a relatively high LD50 value. In an unrelated trial that was prompted by this finding, this isolate was confirmed to be benzimidazole-resistant. Isolate RM8 and an isolate susceptible to all 3 anthelmintics (SK2) were used in the second experiment, which was conducted to monitor changes in the LD50 values of LEV, TBZ and IVM over time following a single infection of 35 000 infective larvae in young sheep. Faecal samples were collected weekly from 24 to 115 DAI. With all 3 anthelmintics, the LD50 values increased with time to a peak around 50-60 DAI, and then declined to levels similar to those observed soon after patency. This trend was consistent for both isolates. The highest mean LD50 values for isolates SK2 for IVM and TBZ and RM8 for IVM and RM8, respectively, were 1.7 and 1.8 times, and 2.2 and 2.9 times higher than the initial mean LD50 values. There was a clear distinction in LD50 values between isolates at each sampling day for both IVM and TBZ. However, as a consequence of the changes in LD50 values with time, the peak LD50 values of IVM for isolate SK2 were higher than the minimum LD50 values of isolate RM8. As there was no apparent difference in LEV efficacy between these 2 isolates, the data were pooled. The highest mean LD50 value was 2.3 times higher than the initial LD50 value. (C) 1997 Australian Society for Parasitology.

Larval stages of Pyromaia tuberculata (Lockington, 1877) (Decapoda, Majidae, Inachinae) reared in the laboratory

The larval development of Pyromaia tuberculata (Lockington, 1877) is described from specimens reared in the laboratory. Larvae were obtained from ovigerous females collected by trawl in Ubatuba, São Paulo state, Brazil. Rearing was carried out at 24 +/- 1 degrees C in 35 parts per thousand S. The larvae pass through two zoeal stages before moulting to the megalopa. A comparison is made with previously studied majid species occurring in the southern and southeastern regions of Brazil.

Electrophoretical studies of proteins of the hypopharyngeal glands and of the larval food of Melipona quadrifasciata anthidioides Lep (Hymenoptera, Meliponinae)

The electrophoretical protein patterns of hypopharyngeal glands, larval food of Melipona, and royal jelly of Apis were compared.Since protein patterns of hypopharyngeal glands from newly emerged workers, brood cell provisioners and foragers are similar to freshly deposited larval food, the identical protein bands probably represent actual gland secretion. This suggests that, as in Apis, the glands secrete proteins to the larval food, and maintain this ability throughout life, although at slightly different intensities, according to the activity of the bees.The similarity on the electrophoretic profiles of the major larval food protein in Apis and Melipona is an interesting finding because of its probable evolutionary significance.

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.

Comparative larval morphology of eight species of Hypsiboas Wagler (Amphibia, Anura, Hylidae) from Argentina and Uruguay, with a review of the larvae of this genus

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

Larval dispersal in Chrysomya megacephala, Chrysomya putoria and Cochliomyia macellaria (Dipt., Calliphoridae): Ecological implications of aggregation behaviour

In this study we investigate aggregated patterns as a consequence of post-feeding larval dispersal in three blowfly species, based on the frequency distribution of sampling units in the substrate having 0, 1, 2,..., n pupae. Statistical analysis revealed that aggregated patterns of distribution emerge as a consequence of larval dispersal, and Cochliomyia macellaria has higher levels of aggregation when compared to Chrysomya megacephala and C. putoria. Aggregation during dispersal is associated with a spatial pattern where most larvae in the species tend to pupariate near the food source. The possible consequences for the population ecology of these species are discussed.

Western blot analysis of tick antigens from a Rhipicephalus sanguineus unfed larval extract and identification of antigenic sites in tick sections using immunohistochemistry. A comparative study between resistant and susceptible host species

Most parasite-host relationships are characterized by the development of resistance by the host, thus limiting the number of parasites. However, some cases are very unusual. In the relationship of the domestic dog with the brown dog-tick Rhipicephalus sanguineus this does not occur, whereas guinea pigs develop efficient resistance. Sera from domestic dogs, crab-eating foxes and guinea pigs collected before and after infestation with R. sanguineus ticks, and after immunization with a whole tick adult or larval homogenate, were used in Western blot analysis to compare and identify potential important antigens from a tick larval homogenate. The same sera were tested in an indirect immunohistochemistry assay in an attempt to compare relevant antigenic sites on histological tick sections. The immunoblotting displayed antigens recognized only by the guinea pigs, as well as several shared antigens between host species, depending on the kind of immunization. Immunohistochemistry revealed probable antigenic sites on the cells and tissues of ticks, which varied depending on the kind of immunization (infestation or vaccination) and the animal species involved.

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.

Diffusion Model Applied to Postfeeding Larval Dispersal in Blowflies (Diptera: Calliphoridae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of temperature on larval survival, development and respiration in Chasmagnathus granulata (Crustacea, Decapoda)

Larvae of an estuarine grapsid crab Chasmagnathus granulata Dana 1851, from temperate and subtropical regions of South America, were reared in seawater (32 ‰) at five different constant temperatures (12, 15, 18, 21, 24 °C). Complete larval development from hatching (Zoea I) to metamorphosis (Crab I) occurred in a range from 15 to 24 °C. Highest survival (60% to the first juvenile stage) was observed at 18°C, while all larvae reared at 12°C died before metamorphosis. The duration of development (D) decreased with increasing temperature (T). This relationship is described for all larval stages as a power function (linear regressions after logarithmic transformation of both D and T). The temperature-dependence of the instantaneous developmental rate (D-1) is compared among larval stages and temperatures using the Q10 coefficient (van't Hoff's equation). Through all four zoeal stages, this index tends to increase during development and to decrease with increasing T (comparing ranges 12-18, 15-21, 18-24 °C). In the Megalopa, low Q10 values were found in the range from 15 to 24 °C. In another series of experiments, larvae were reared at constant 18°C and their dry weight (W) and respiratory response to changes in T were measured in all successive stages during the intermoult period (stage C) of the moulting cycle. Both individual and weight-specific respiration (R, QO2) increased exponentially with increasing T. At each temperature, R increased significantly during growth and development through successive larval stages. No significantly different QO2 values were found in the first three zoeal stages, while a significant decrease with increasing W occurred in the Zoea IV and Megalopa. As in the temperature-dependence of D, the respiratory response to changes in temperature (Q10) depends on both the temperature range and the developmental stage, however, with different patterns. In the zoeal stages, the respiratory Q10 was minimum (1.7-2.2) at low temperatures (12-18 °C), but maximum (2.2-3.0) at 18-24 °C. The Megalopa, in contrast, showed a stronger metabolic response in the lower than in the upper temperature range (Q10 = 2.8 and 1.7, respectively). We interpret this pattern as an adaptation to a sequence of temperature conditions that should typically be encountered by C. granulata larvae during their ontogenetic migrations: hatching in and subsequent export from shallow estuarine lagoons, zoeal development in coastal marine waters, which are on average cooler, return in the Megalopa stage to warm lagoons. We thus propose that high metabolic sensitivity to changes in temperature may serve as a signal stimulating larval migration, so that the zoeae should tend to leave warm estuaries and lagoons, whereas the Megalopa should avoid remaining in the cooler marine waters and initiate its migration towards shallow coastal lagoons.

Theoretical approaches to forensic entomology: II. Mathematical model of larval development

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

Larval predation by Chrysomya albiceps on Cochliomyia macellaria, Chrysomya megacephala and Chrysomya putoria

Chrysomya albiceps, the larvae of which are facultative predators of larvae of other dipteran species, has been introduced to the Americas over recent years along with other Old World species of blowflies, including Chrysomya megacephala, Chrysomya putoria and Chlysomya rufifacies. An apparent correlate of this biological invasion has been a sudden decline in the population numbers of Cochliomyia macellaria, a native species of the Americas. In this study, we investigated predation rates on third instar larvae of C. macellaria, C. putoria and C. megacephala by third instar larvae of C. albiceps in no-choice, two-choice and three-choice situations. Most attacks by C. albiceps larvae occurred within the first hour of observation and the highest predation rate occurred on C. macellaria larvae, suggesting that C. albiceps was more dangerous to C. macellaria than to C. megacephala and C. putoria under these experimental conditions. The rates of larvae killed as a result of the predation, as well as its implications to population dynamics of introduced and native species are discussed.

Larval aggregation and competition for food in experimental populations of Chrysomya putoria (Wied.) and Cochliomyia macellaria (F.) (Dipt., Calliphoridae)

In blowflies, larval aggregation in patches of food can be both intra- and interspecific, depending upon the degree to which competitors are clumped among the patches. In the present study, the implications of spatial aggregation for larval competition was investigated in experimental populations of the introduced blowfly Chrysomya putoria and the native Cochliomyia macellaria, using data from survival to adulthood in a range of single- and double-species larval cultures. The reduction in C. macellaria survival rate in the presence of C. putoria suggests that the former species is the inferior competitor. The results on survival to adulthood for both species in single- and double-species cultures can be explained in the light of the relationship between the level of intra- and interspecific aggregation and the efficiency of the larval feeding process. The possible implications of these results for the population biology of both species in natural environments are discussed.

Larval development of Epialtus bituperculatus H. Milne Edwards, 1834 (Crustacea: Decapoda: Brachyura: Majidae) with comments on majid larvae from the southwestern Atlantic

The larval development of the spider crab Epialtus bituberculatus H. Milne Edwards which lives on rocky shores with algae such as Sargassum and Hypeneia, is described. Larvae were obtained from ovigerous females collected in Ubatuba, State of São Paulo, Brazil. Rearing was carried out at 24 ± 1°C, with an average salinity of 35%. Larval development consists of two zoeal stages and one megalopa. Zoeal development was completed in 9.5 days. Analysis indicated that zoeae of E. bituberculatus are very similar to those of E. brasiliensis Dana and Acanthonyx scutiformis (Dana). Differences noted between these species pertain to the setation of the carapace, maxillule and second maxilliped. The main morphological features useful for identification are presented together with a summary of features that characterize larvae of majid subfamilies in Brazil. A key for the identification of southwestern Atlantic majid zoeae to the family level is provided.