Morphological regional differences of epithelial cells along the midgut in Diatraea saccharalis Fabricius (Lepidoptera : Crambidae) larvae

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

The influence of initial feeding on muscle development and growth in pacu Piaractus mesopotamicus larvae

The effects of different feeding schemes on pacu Piaractus mesopotamicus early development were evaluated with respect to growth, survival, muscle development, and differential gene expression of MyoD and myogenin. The pacu larvae (4 days post hatch-dph, 0.77 mg wet weight) were given six feeding treatments intentionally designed to cause variations in the larvae growth rate: (A) only artemia nauplii; (CD) only a commercial diet; (ED) only a semi-purified experimental diet; (ACD) and (AED) two treatments that involved weaning; and (S) starvation. Early weaning from artemia nauplii to the formulated diets (ACD and AED) affected growth and survival of the pacu larvae compared with the exclusive use of artemia (A). Starvation (S) and the commercial diet (CD) caused total mortality in pacu larvae at 18 dph. The experimental diet (ED) assured low fish survival and growth. The skeletal muscle morphology was not affected by the delay in somatic growth from early weaning onto the formulated diets. Three distinct muscle compartments were observed throughout the larval development in treatments A, ACD and AED: superficial, deep and intermediate, accompanied by muscle thickening. Severe undernourishment caused drastic differences in growth and in the morphology of the muscle fibers. Pacu larvae fed only formulated diets (CD and ED) showed muscle characteristics similar to the larvae in starvation (S) during the first 15 dph. At 27 and 35 dph, a slight increase in epaxial muscle mass was noted in larvae fed only the experimental diet (ED). At 35 dph, we observed a high frequency of fibers >= 40 mu m in the larvae that were weaned onto the formulated diets (ACD and AED), indicative of hypertrophy. In contrast, the larvae fed only artemia nauplii (A) displayed a larger number of fibers with diameters <= 20 mu m, which is indicative of hyperplasia. The expression of the MyoD and myogenin genes in pacu larvae at 35 dph was not affected by initial feeding (p>0.05). In conclusion, the formulated diets used impaired pacu larvae growth and survival; therefore, they were inadequate for pacu, at least at the times they were introduced. Artemia nauplii were the most adequate food source during first feeding of the pacu, and they produced bigger fish upon completion of the experiment. Moreover, the contribution of hyperplasia to the skeletal muscle growth appeared higher in fast- than in slow-growing pacu larvae. (C) 2011 Elsevier By. All rights reserved.

Ultramorphology of digestive tract of Anticarsia gemmatalis (Hubner, 1818) (Lepidoptera : Noctuidae) at final larval development

The digestive tract of insects is an important natural, physical, and chemical defense barrier against pathogen invasion. Certain. lepidopteran caterpillars are serious pests of agricultural crops and their biology has received much attention, but little is known about the larval noctuid gut. The morphological analysis of the digestive tract in Anticarsia gemmatalis under scanning electron microscopy (SEM) is a good model for studies about its defense mechanism. The material was fixed (2,5% glutaraldehyde solution; 0.1 M-phosphate buffer, pH 7.3), post-fixed (1% osmium tetroxide in the same buffer), dried at critical point, gold coated and analyzed in a SEM 515-Philips. A. gemmatalis digestive tract consists of a straight duct of varying length and diameter, subdivided in three main regions: the foregut formed by the oral cavity, pharynx, esophagus, and crop; the midgut that is the largest portion of the digestive tract without noticeable morphological differentiation along its length; and the hindgut that is morphologically differentiated in pylorus, ileum, colon, and rectum. Although the general morphology of the A. gemmatalis digestive tract is quite similar to the other Lepidoptera species, the anatomical array of the crop muscular layers is quite different comparing with the description for other larval insect.

Dispersal and predation behavior in larvae of Chrysomya albiceps and Chrysomya megacephala (Diptera : Calliphoridae)

Chrysomya albiceps and Chrysomya megacephala are exotic blowfly species known by producing myiasis in humans and other animals and by transmitting pathogens mechanically. C. albiceps stand out by being a facultative predator of other dipteran larvae. In this paper we investigated the influence of larval predation on the dispersal of larvae of C. albiceps and C. megacephala single and double species for three photophases. An experimental acrylic channel graduated and covered with wood shavings was used to observe the larval dispersal. The results showed that C. albiceps attacks C. megacephala larvae during dispersal and keeps an aggregated pattern close to the release point, in single and double species, independently of the different photophases. Chrysomya megacephala single species exhibited the same pattern, but in double species this was changed to a random distribution.

Theoretical estimates of consumable food and probability of acquiring food in larvae of Chrysomya putoria (Diptera: Calliphoridae)

An indirect estimate of consumable food and probability of acquiring food in a blowfly species, Chrysomya putoria, is presented. This alternative procedure combines three distinct models to estimate consumable food in the context of the exploitative competition experienced by immature individuals in blowfly populations. The relevant parameters are derived from data for pupal weight and survival and estimates of density-independent larval mortality in twenty different larval densities. As part of this procedure, the probability of acquiring food per unit of time and the time taken to exhaust the food supply are also calculated. The procedure employed here may be valuable for estimations in insects whose immature stages develop inside the food substrate, where it is difficult to partial out confounding effects such as separation of faeces. This procedure also has the advantage of taking into account the population dynamics of immatures living under crowded conditions, which are particularly characteristic of blowflies and other insects as well.

Prey choice by facultative predator larvae of Chrysomya albiceps (Diptera: Calliphoridae)

In this study we investigated predation rates on third instar larvae of Chrysomya putoria and C. megacephala by third instar larvae of C. albiceps in a two-choice situation. The highest predation rate occurred on C. putoria larvae and this result is compared to previous experiments, in which C. macellaria larvae were present. Our results suggest that, when C. macellaria is absent C. albiceps larvae attack more C. putoria than C. megacephala larvae. Prey choice decisions and its implications for introduced and native blowflies are discussed.

Vertical migration of Haemonchus contortus third stage larvae on Brachiaria decumbens grass

The present study aimed at evaluating the vertical migration of Haemonchus contortus third stage larvae (1-3) on Brachiaria decumbens grass, as well as at verifying whether larval numbers on pasture varies over the day due to climatic conditions. Feces containing H. contortus L3 were deposited on the soil in the middle of herbage which was initially 30 cm high. Seven days later, samples of different herbage strata (0-10, 10-20 and >20 cm), remaining feces and a layer of approximately 1 cm soil were collected. Tests were carried out in four periods: September 2006, December 2006, March 2007, and June 2007. Samples were collected at sunrise, mid-day, sunset, and mid-night. The humidity and temperature conditions observed in different months influenced larval migration from the feces to the grass. In September, December and March, it rained after fecal deposition on pasture, which favored migration of larvae from the feces to the herbage. Conversely, in June 2007, when there was no rainfall after fecal deposition and temperatures were lower, L3 were mainly recovered from feces. As regards the vertical migration of larvae, the numbers of H. contortus L3 in the forage strata remained relatively constant over the day. This indicates there is not a determined period in which sheep on pasture are at higher risk of infection. Finally, in all collection periods a considerable amount of third stage larvae was observed on the herbage top, which is the first plant part consumed by sheep. (C) 2008 Elsevier B.V. All rights reserved.

Characterization of the excretory/secretory products of Dermatobia hominis larvae, the human bot fly

Proteolytic activity in excretory/secretory products (ESP) of first- (L1), second- (L2) and third-instar (L3) larvae of Dermatobia hominis was analyzed through gelatin-gel and colorimetric enzyme assays with the chromogenic substrates azocasein and BApNA. The functional characterization of proteases was based on inhibition assays including synthetic inhibitors. ESP were obtained from new-hatched larvae reared in the laboratory and from second- and third-instar larvae removed from naturally infested cattle. Gelatin-gel analysis evidenced few bands of proteolysis, predominantly of high apparent molecular masses, in ESP of L1, whereas in the gel of L2 and U ESP there was a wide range of proteolytic activity most of them not resolved in a single species. Azocasein assays revealed a progressive increase of protease activity from first- to third-instar larvae. Protease inhibitor assays revealed a predominance of metalloproteases in L1 ESP that could be related to a skin penetration process and to a diversion of host immune response. The predominance of serine proteases in L2 and L3 and the great tryptic activity presented by L3 ESP were attributed to an increasing trophic activity by the growing larvae, since the viability of adult flies strictly depends on larval abilities to assimilate nutrients from the host. Taking together, these results suggest that Dematobia larvae secrete/excrete different proteases that may be related to diverse functions during host penetration and infestation, which reinforces the relevance of the study of such proteolytic enzymes. (C) 2009 Elsevier B.V. All rights reserved.

The first zoeal stage of two species of Grapsidae (Decapoda, Brachyura) and a key to such larvae from the Brazilian coast

First zoeal stages of the grapsinid Goniopsis cruentata (Latreille, 1803) and the sesarminid Aratus pisonii (H. Milne Edwards, 1837), are described and illustrated. Grapsinae zoeae can be distinguished from the other grapsid larvae by the absence of lateral spines on the carapace and the reduction of the antennal exopod to a small seta. A key to the first zoeal stage of the Brazilian coast Grapsidae is provided.

Larval development of Epialtus bituberculatus 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 Hypneia, is described. Larvae were obtained from ovigerous females collected in Ubatuba, State of São Paulo, Brazil. Rearing was carried out at 24 +/- 1 degreesC, with an average salinity of 35 parts per thousand. 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.

Ultrastructure of the midgut epithelium of Meliponinae larvae with different developmental stages and diets

The comparative study of the ultrastructure of the midgut epithelium of stingless bee larvae that eat plant protein (pollen) and animal protein (carrion) throughout the larval phase, shows variations in the digestive cells that are only relative to larval aging and not to the type of larval diet. The cells of older larvae present a cytoplasm with empty spaces that result from emptying of lipid and glycogen stocks, and the presence of autophagic vacuoles. These results are discussed in relation to the hypothesis that variations in the digestive tract of insects may be associated with different diets or phylogeny. We conclude that different diets do not determine cell morphology adaptations in the studied species. As the variations in the ultrastructure of the midgut epithelium are the same in all studied species, including the necrophagous species Trigona hypogea, throughout the larval stage, this sequence of changes seems to be due to different physiological state during larval development.

Differentiation of the worker's ovary in Apis mellifera L. (Hymenoptera, Apidae) during life of the larvae

The aim of the present study is to characterize the way worker and queen ovaries differentiate in, Apis mellifera, a species with trophic determination of female castes. A morphological study carried out with light and transmission electron microscopy showed that the differences in ovary development between the two castes begin as soon as the differential nursing of larvae is initiated. The decrease in ovariole number in worker ovaries is due to a process of cell death occurring in germinative cells and autophagic regression of somatic cells in the ovarioles that commence in the third instar larvae and proceed until the fifth instar where the process is more intense. Germinative cell death leads to ovariole disintegration and incorporation of the remaining somatic cells of the latter into the stromatic cells in such a way that the total volume of the ovary is little affected during larval development, although the ovariole number decreases. By the end of the larval stage, loss of cells is observed among the stromatic cells of the ovary. As a result, the ovary starts to decrease in volume and takes on the adult form.

In situ localization of heat-shock proteins and cell death labelling in the salivary gland of acaricide-treated honeybee larvae

The effects of the acaricides, rotenone and oxalic acid (OA) on salivary glands of honeybee larvae were evaluated. Immunohistochemical methods were used to detect cell death and heat-shock protein (HSP70 and 90) localizations. Heat-shock proteins (HSP70 and 90) were localized in the cytoplasm and/or the nuclei of secretory gland cells, both under stress and in normal conditions. In rotenone-treated larvae, there were no changes in the normal level of cell death and also there were no morphological alterations in the secretory cells. In the larvae treated with oxalic acid, the salivary gland showed varying degrees of morphological cellular alteration and an increase in the cell death level. The present data suggest that stress-induced HSP70 might have an antiapoptotic effect while the stress-induced HSP90 might have a chaperone function in the larval salivary glands.

Lipid distribution in salivary glands of larvae and adult bees (Hymenoptera : Apidae)

Cytochemical studies were carried out to establish lipid distribution in the salivary glands of larvae and adult bees, using the imidazole buffer technique. In the duct cells of the larval salivary gland, the reaction was positive in the epicuticle and negative in the glandular lumen. The absence of smooth endoplasmic reticulum and the presence of lipids in the intercellular space suggest that lipids absorbed from the haemolymph could be used in the constitution of the epicuticle, after having been conveyed through the epithelium. In adult workers (new-emerged, nurse and forager workers), the head salivary glands presented a positive reaction in the secretion in glandular lumen, identifying its lipidic nature.