On the larval stages of Macrocoeloma diplacanthum (Decapoda : Brachyura : Majidae), and a review of mithracine phylogenetic aspects

Larval development of Macrocoeloma diplacanthum (Stimpson) consists of two zoeal stages, followed by the megalopa. Each larval stage is described in detail. The duration of the zoeal stages was 2-3 (2.4 +/- 0.5) and 3-4 (3.5 +/- 0.5) days for the first and second zoea, respectively, the megalopa phase appearing 6-8 (7.0 +/- 0.5) days after hatching. Unlike for other majids, zoeal stages of M. diplacanthum can be readily distinguished by their distended forehead with strong underlying muscle bands, undercut dorsal carapace spine, and spine on the terminal endopod segment of the first maxilliped. No other known mithracine or majid zoeae exhibit this combination of features. Our zoeal account of M. diplacanthum from Mexico is remarkably consistent with Floridian specimens previously described. However, we have found some differences between descriptions, which could be attributed to natural variation or inadequate description. Previous attempts to evaluate the relationships within Mithacinae have been based on larval characters widely distributed throughout Majidae and therefore are considered inadequate to infer sister-group relationships. The phylogenetic analysis of majids suggested that the position of Mithracinae is still uncertain, as is its monophyletic status. We recommend that additional characters, particularly of the megalopa phase, be sought for a better resolution of majid evolutionary history.

Searching for larval support for majoid families (Crustacea : Brachyura) with particular reference to Inachoididae Dana, 1851

We re-evaluated the larval support for families within majoids using the Wilcoxon signed-rank test with emphasis on Inachoididae. To accomplish our objectives, we added 10 new taxa, two of which are traditionally assigned to the family of special interest, to a previous larval database for majoids, and re-appraised the larval characters used in earlier studies. Phylogenetic analysis was performed with PAUP* using the heuristic search with 50 replicates or the branch-and-bound algorithm when possible. Multi-state transformation series were considered unordered; initially characters were equally weighted followed by successive weighting, and trees were rooted at the Oregoniidae node. Ten different topological constraints were enforced for families to evaluate tree length under the assumption of monophyly for each taxonomic entity. Our results showed that the tree length of most constrained topologies was not considerably greater than that of unconstrained analysis in which most families nested as paraphyletic taxa. This may indicate that the present larval database does not provide strong support for paraphyly of the taxa in question. For Inachoididae, although the Wilcoxon signed-rank test rejected a significant difference between unconstrained and constrained cladograms, we were unable to provide a single synapomorphy for this clade. Except for the conflicting position of Leurocyclus and Stenorhynchus, the two clades correspond to the traditional taxonomic arrangement. Among inachoidids, the clade (Anasimus (Paradasygyius (Collodes + Pyromaia))) is supported, whereas for inachids, the clade (Inachus (Macropodia + Achaeus)) is one of the most supported clades within majids. As often stated, only additional characters will provide a better test for the monophyly of Inachoididae and other families within Majoidea.

Larval development of Apiomithrax violaceus (A. Milne Edwards, 1868) (Decapoda : Brachyura : Majoidea : Pisidae) reared in laboratory conditions, and a review of larval characters of Pisidae

Apiomithrax violaceus (A. Milne Edwards, 1868 ) is a pisid majoid crab occurring in tropical and subtropical coastal waters of the eastern and western South Atlantic. Larval development consists of two zoeal stages and a megalopa. Beginning with the first zoea, the duration of each larval stage at 24degreesC was 3-8 (5+/-1), 3-5 (4+/-0.5) and 9-15 (11+/-2) days, the megalopa and first crab instar appearing 9-11 (10+/-1) and 20-27 (23+/-2) days after hatching, respectively. Larval characters agree with those proposed for the Majoidea, in having nine or more setae on the scaphognathite in the first zoea and well-developed pleopods in the second zoea. However, larvae of A . violaceus do not fit larval pisid features. Zoeal stages differ from most other Pisidae in having lateral spines, a long rostral spine extending beyond the antenna, two spines per telson fork and a dorsolateral process on the third abdominal somite. The megalopa differs in having a spine dorsally on the carapace and on the basial segment of the second pereiopod. Two characters that are potentially unique to Apiomithrax include a zoeal antenna with an exopod that is much longer than the protopod, and a rostral spine that is longer than the dorsal spine. These characters should facilitate the identification of this taxon and could also be useful for phylogenetic studies. A review of larvae of 28 species among 14 genera indicated that there is no apparent single larval character that differentiates the Pisidae, with more limited phylogenetic analyses suggesting that this is a paraphyletic group. Apiomithrax , Eurynolambrus , Pisoides , Rochinia and Scyra have the most divergent morphological characters within the family. The analysis and inclusion of additional taxa is likely to shed more light on the sister-group relationships of the Pisidae. However, based on the extent of morphological interspecific variability of known larvae it is likely that the group, as presently defined by adult morphology, is not monophyletic.

Larval cannibalism rates in the mangrove crab Ucides cordatus (Decapoda : Ocypodidae) under laboratory conditions

Predation of zoeas by megalopae of Ucides cordatus is frequently observed in the laboratory during larval rearing, a phenomenon that could considerably reduce the output of larviculture. Experiments were carried out in the present study to assess how the survivorship of larvae at the end of the larviculture is influenced by cannibalism by megalopae on the larvae of earlier stages, as well as on other megalopae. In addition, tests were performed to assess whether the adoption of different feeding protocols can decrease cannibalism rates. Experiments were carried out in plastic vials containing ocean water (salinity 25 g L-1) under controlled environmental conditions (26 degrees C and 16:8 h LD photoperiod). An ensemble analysis of all the developmental stages indicated that zoeal mortality rates were significantly higher in the presence of megalopae, a result that is consistent with cannibalism by megalopae. However, separate analysis for each developmental stage indicated that only zoea IV, V and VI show reduced survivorship. No cannibalism was detected among megalopae. Food supplementation using Artemia sp. at a density of 6 nauplii mL(-1) proved to be successful in reducing cannibalism rates, whereas supplementation at a lower density (0.3 nauplii mL(-1)) failed to show such an effect. The implications of these results for the larviculture of U. cordatus are discussed.

FIRST ZOEAL STAGE and MEGALOPA of UCA (UCA) MARACOANI (DECAPODA: BRACHYURA), WITH COMMENTS on THE LARVAL MORPHOLOGY of SOUTH-AMERICAN SPECIES of OCYPODIDAE

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

Ultrastructure of last larval instar fat body cells of Pachycondyla (= Neoponera) villosa (Formicidae : Ponerinae): cytochemical and chemical analysis

The ultrastructure of the fat body cells (trophocytes) of the last larval instar of Pachycondyla (= Neoponera) villosa is presented. The cytoplasm is restricted to the cell periphery and to the smaller strips among the vacuoles, protein granules, lipid droplets, and around the nucleus. Cytochemically, the presence of basic amino acids in the protein granules and in the nuclei was observed by using the ethanolic phosphotungstic acid technique (EPTA). The lipid droplets stained for unsaturated lipids. This result was further confirmed by gas chromatography and mass spectrometry, where the unsaturated fatty acids were identified as oleic and linoleic acids together with saturated fatty acids such as palmitic and stearic acid. Carbohydrates (glycogen) were also detected in the fat body. The glycogen is present as beta particles distributed among the lipid droplets and sometimes attached to them.

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

Ultramorphology and histology of the larval salivary gland of Pachycondyla villosa (Fabricius) (Hymenoptera: Formicidae, Ponerinae)

A glândula salivar apresenta-se com um duto anterior único, formado por um epitélio colunar, dois dutos laterais curtos, os quais apresentam-se com epitélio cúbico simples e que na sua porção mais proximal torna-se colunar. Posterior a estes, encontram-se os dois reservatórios, os quais possuem o epitélio bastante delgado e é neste reservatório que a região secretora da glândula se abre. Os ramos dorsal e ventral da região secretora da glândula conectam-se por meio de comissuras transversais, sendo que, posteriormente, a região secretora termina em forma de alça. A região secretora é uniforme, não apresenta tipos celulares distintos e é formada por um epitélio cúbico simples. Neste trabalho é apresentada, também, a revisão sobre a morfologia da glândula salivar larval em insetos, principalmente com relação aos Hymenoptera-Aculeata.

Nuclear alterations associated to programmed cell death in larval salivary glands of Apis mellifera (Hymenoptera : Apidae)

The silk glands of bees are a good model for the study of cell death in insects. With the objective to detect the nuclear features during glandular regression stage, larvae at the last instar and pre-pupae were collected and their silk glands were dissected and processed for ultrastructural analysis and histologically for cytochemical and imunocytochemical analysis. The results showed that the cellular nuclei exhibited characteristics of death by atypical apoptosis as well as autophagic cell death. Among the apoptosis characteristic were: nuclear strangulation with bleb formation in some nuclei, DNA fragmentation in most of the nuclei and nucleolar fragmentation. Centripetal chromatin compaction was observed in many nuclei, forming a perichromatin halo differing from typical apoptotic nuclei. With regards to the characteristics of autophagic-programmed cell death, most relevant was the delay in the collapse of many nuclei. (C) 2006 Elsevier Ltd. All rights reserved.

Effect of larval food amount on ovariole development in queens of Trigona spinipes (Hymenoptera, Apinae)

Caste determination in Trigona spinipes Fabricius (Hymenoptera, Apidae, Meliponini) is trophogenic. Larvae that eat about 360 mu l of food become queens, while those who consume 36 mu l develop into workers. We studied the effect of larval nutrition on the number and length of ovarioles and on ovarian development in fifth instar larvae, white eyed, pink eyed and black-eyed pupae as well as newly emerged adults. All larvae have four ovarioles per ovary, while in queen pupae this number ranged from 8 to 15. Cyst formation, the cell death and other characteristics of ovary morphogenesis were the same regardless of the quantity of food consumed. These results are discussed in relation to caste differentiation in other bees.

Morphological and biochemical analysis of the stored and larval food of an obligate necrophagous bee, Trigona hypogea

Trigona hypogea, T. crassipes, and T. necrophaga are obligate necrophagous bees that differ from the majority of bees by using animal material instead of pollen as a protein resource. Since T. hypogea does not store protein in cerumen pots, it was thought that glandular secretions were its only larval protein source. This is in contrast to T. necrophaga which stores a yellowish proteinaceous jelly in the pots. Our results show that the larval food of T. hypogea has a higher protein content than the food stored in the pots and that it presents an electrophoretical protein pattern similar to that of the hypopharyngeal gland, indicating that workers add glandular secretions to the larval food while provisioning the brood cells. Thus, it can be suggested that T. hypogea has a provisioning behavior similar to other Meliponinae. The presence of several bands of proteins in the food stored in the pots shows that this species stores carrion mixed with honey in storage pots. Morphological data suggest that both larvae and adults make use of the same foodstuffs. These results also suggest that T. hypogea is more similar to other necrophagous species than it is to T. necrophaga (a more specialized bee).

Ant predation of larval Diatraea saccharalis Fab. (Lep., Crambidae) in new sugarcane in Brazil

Ant predation of natural field populations of larval sugarcane borers, Diatraea saccharalis, was evaluated by using insecticidal check techniques, and through survivorship studies of artificial high-density infestations, in plantations in Brazil. Areas of ant suppression had higher levels of D. saccharalis than unsuppressed areas, although both types of areas normally had extremely low D. saccharalis populations. Artifical infestations of D. saccharalis yielded significantly lower larval survivorship in unsuppressed areas than in suppressed areas. Fire ants (Solenopsis spp.) were always the most abundant ant. This study has demonstrated that ants are important pest predators in Brazilian sugarcane fields.

Larval ecomorphology of 13 Libellulidae (Anisoptera, Odonata) of the Middle Rio Doce Valley, Minas Gerais, Brazil

Nos lagos do Médio Rio Doce (MG), dois grupos de larvas de Libellulidae apresentam diferenças no uso do habitat: um utiliza preferencialmente macrófitas aquáticas e o outro utiliza a superfície do fundo. O objetivo deste trabalho foi verificar se existe uma distinção morfológica entre estes dois grupos. Treze medidas morfológicas foram tiradas das larvas e analisadas. Nenhuma diferença, com relação ao tamanho corporal, foi encontrada entre os dois grupos, embora diferenças de forma tenham sido observadas com relação a duas variáveis morfológicas. As espécies que usam principalmente macrófitas tendem a possuir maiores medidas relativas do lábio e menores medidas da largura do abdome, o oposto ocorrendo com as larvas habitantes do fundo. Vantagens na captura de presas e na vulnerabilidade à predação são provavelmente as explicações para a divergência morfológica observada entre os dois grupos de larvas.

A review of postfeeding larval dispersal in blowflies: implications for forensic entomology

Immature and adult stages of blowflies are one of the primary invertebrate consumers of decomposing animal organic matter. When the food supply is consumed or when the larvae complete their development and migrate prior to the total removal of the larval substrate, they disperse to find adequate places for pupation, a process known as postfeeding larval dispersal. Several important ecological and physiological aspects of this process were studied since the work by Green (Ann Appl Biol 38:475, 1951) 50 years ago. An understanding of postfeeding larval dispersal can be useful for determining the postmortem interval (PMI) of human cadavers in legal medicine, particularly because this interval may be underestimated if older dispersing larvae or those that disperse longer, faster, and deeper are not taken into account. In this article, we review the process of postfeeding larval dispersal and its implications for legal medicine, in particular showing that aspects such as burial behavior and competition among species of blowflies can influence this process and consequently, the estimation of PMI.

Larval competition for patchy resources in Chrysomya megacephala (Dipt., Calliphoridae): implications of the spatial distribution of immatures

In the present study, a single procedure was established to investigate the effect of the spatial distribution of immatures in patchy resources, on the outcome of larval competition for food. in experimental populations of Chrysomya megacephala. A theoretical model of intraspecific competition was extended and applied to experimental data on survival to adulthood for 20 larval densities, to obtain the theoretical mean number of individuals that will survive, considering a hypothetical previous random adult oviposition in a system of homogeneous patches. The survival curve obtained suggests that the larval competition for food in C. megacephala is of the scramble/exploitative type, which corroborates results from previous studies, although the latter did not consider the correlation between local and global abundances. The present model allows that experimental data could be perfectly applicable, and it incorporates fundamental assumptions about the spatial context of competition for patchy resources in blowflies, and may be applied to the optimization of mass rearing techniques and to the maintenance of insect colonies under experimental conditions.