On the Relationship between Zaprionus spinipilus Chassagnard & McEvey and Z. vittiger Coquillett, the Type Species of the Genus Zaprionus (Diptera: Drosophilidae)

Zaprionus vittiger Coquillett is the type species of the genus Zaprionus Coquillett. However, the species is only known from five old museum specimens collected from South Africa and Malawi. It has often been confused with many other Zaprionus species, especially with Z. spinipilus Chassagnard & McEvey, a widespread species in Africa known from Madagascar, Malawi, Ethiopia and Cameroon. We have recently collected flies from the type localities of both species (South Africa and Madagascar, respectively). This has prompted us to test the taxonomic boundaries of these two nominal species using molecular (the mitochondrial COII and the nuclear Amyrel genes), chromosomal, morphological (internal and external genitalia), and reproductive isolation analyses. The results suggest Z. spinipilus to be a junior synonym to Z. vittiger.

Genetic structure of Partamona helleri (Apidae, Meliponini) from Neotropical Atlantic rainforest

Stingless bees of the genus Partamona are distributed from southern Mexico to southern Brazil. This genus has been subject to different approaches to solve questions concerning general biology, taxonomy, systematics and biogeography, but population studies applying molecular techniques are inexistent. We analyzed the genetic structure of P. helleri across its geographic distribution along the coastal Atlantic tropical rainforest in Brazil. Ten mtDNA haplotypes were observed in 47 colonies of P. helleri of which some were exclusive and others shared among geographic sub-groups. Statistical analysis showed high genetic differentiation between geographic areas sampled. Fragmentation of the Atlantic forest during Pleistocene glaciations is discussed as a possible cause of the present haplotype distribution and frequency.

Digestive morphophysiology of Gryllodes sigillatus (Orthoptera: Gryllidae)

The evolution of the digestive system in the Order Orthoptera is disclosed from the study of the morphophysiology of the digestive process in its major taxa. This paper deals with a cricket representing the less known suborder Ensifera Most amylase and trypsin activities occur in crop and caeca. respectively. Maltase and aminopeptidase are found in soluble and membrane-bound forms in caeca, with aminopeptidase also occurring in ventriculus. Amaranth was orally fed to Gryllodes sigillatus adults or injected into their haemolymph. The experiments were performed with starving and feeding insects with identical results. Following feeding of the dye the luminal side of the most anterior ventriculus (and in lesser amounts the midgut caeca) became heavily stained. In injected insects, the haemal side of the most posterior ventriculus was stained This suggested that the anterior ventriculus is the main site of water absorption (the caeca is a secondary one). whereas the posterior ventriculus secretes water into the gut. Thus, a putative counter-current flux of fluid from posterior to anterior ventriculus may propel digestive enzyme recycling. This was confirmed by the finding that digestive enzymes are excreted at a low rate. The fine structure of midgut caeca and ventriculus cells revealed that they have morphological features that may be related to their involvement in secretion (movement from cell to lumen) and absorption (movement from lumen to cell) of fluids. Furthermore, morphological data showed that both merocrine and apocrine secretory mechanisms occur in midgut cells. The results showed that cricket digestion differs from that in grasshopper in having (1) more membrane-bound digestive enzymes; (2) protein digestion slightly displaced toward the ventriculus; (3) midgut fluxes, and hence digestive enzyme recycling, in both starved and fed insects. (C) 2009 Elsevier Ltd. All rights reserved.

Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut

Musca domestica larvae display in anterior and middle midgut contents, a proteolytic activity with pH optimum of 3.0-3.5 and kinetic properties like cathepsin D. Three cDNAs coding for preprocathepsin D-like proteinases (ppCAD 1, ppCAD 2, ppCAD 3) were cloned from a M. domestica midgut cDNA library. The coded protein sequences included the signal peptide, propeptide and mature enzyme that has all conserved catalytic and substrate binding residues found in bovine lysosomal cathepsin D. Nevertheless, ppCAD 2 and ppCAD 3 lack the characteristic proline loop and glycosylation sites. A comparison among the sequences of cathepsin D-like enzymes from some vertebrates and those found in M. domestica and in the genomes of Aedes aegypti, Drosophila melanogaster, Tribolium castaneum, and Bombyx mori showed that only flies have enzymes lacking the proline loop (as defined by the motif: DxPxPx(G/A)P), thus resembling vertebrate pepsin. ppCAD 3 should correspond to the digestive cathepsin D-like proteinase (CAD) found in enzyme assays because: (1) it seems to be the most expressed CAD, based on the frequency of ESTs found. (2) The mRNA for CAD 3 is expressed only in the anterior and proximal middle midgut. (3) Recombinant procathepsin D-like proteinase (pCAD 3), after auto-activation has a pH optimum of 2.5-3.0 that is close to the luminal pH of M. domestica midgut. (4) Immunoblots of proteins from different tissues revealed with anti-pCAD 3 serum were positive only in samples of anterior and middle midgut tissue and contents. (5) CAD 3 is localized with immunogold inside secretory vesicles and around microvilli in anterior and middle midguit cells. The data support the view that on adapting to deal with a bacteria-rich food in an acid midgut region, M. domestica digestive CAD resulted from the same archetypical gene as the intracellular cathepsin D, paralleling what happened with vertebrates. The lack of the proline loop may be somehow associated with the extracellular role of both pepsin and digestive CAD 3. (C) 2009 Elsevier Ltd. All rights reserved.

Morphometrical, biochemical and molecular tools for assessing biodiversity. An example in Plebeia remota (Holmberg, 1903) (Apidae, Meliponini)

We see today many efforts to quantify biodiversity in different biomes. It is very important then to develop and to apply other methodologies that allow us to assess biodiversity. Here we present an example of application of three tools with this goal. We analyzed two populations of Plebeia remota from two distinct biomes that already showed several differences in morphology and behavior. Based on these differences, it has been suggested that the populations of Cunha and Prudentopolis do not represent a single species. In order to verify the existence or absence of gene flow between these two groups, we characterized the patterns of mtDNA through RFLP, the patterns of wing venation through geometric morphometry, and the cuticular hydrocarbons through gas chromatography-mass spectrometry. We used bees collected in these two locations and also from colonies which have being kept for around 9 years at Sao Paulo University. We found six different haplotypes in these specimens, of which three of them occurred exclusively in the population of Cunha and three only in the Prudentopolis population. The fact that the populations do not share haplotypes suggests no maternal gene flow between them. The two populations were differentiated by the pattern of the wing veins. They also had different mixtures of cuticle hydrocarbons. Furthermore it was shown that the colonies kept at the university did not hybridize. These two groups may constitute different species. We also show here the importance of using other methodologies than traditional taxonomy to assess and understand biodiversity, especially in bees.

The DNA puff 4C expresses a salivary secretion protein in Trichosia pubescens (Diptera; Sciaridae)

DNA puffs are genomic regions of polytene chromosomes that undergo developmentally controlled DNA amplification and transcription in salivary glands of sciarid flies. Here, we tested the hypothesis that DNA puff genes code for salivary proteins in Trichosia pubescens. To do that, we generated antibodies against saliva and immunoscreened a cDNA library made from salivary glands. We isolated clones corresponding to DNA puff regions, including clone D-50 that contained the entire coding sequence of the previously isolated C4B1 gene from puff 4C. Indeed, we showed that puff 4C is a DNA puff region detecting its local transcription and its extra rounds of DNA incorporation compared to neighboring regions. We further confirmed D-50 clone identity in Western blots reacted with the anti-saliva anitiserum. We detected a recombinant protein expressed by this clone that had the expected size for a full-length product of the gene. We end with a discussion of the relationship between DNA puff genes and their products.

Caves as a Winter Refuge by a Neotropical Harvestman (Arachnida, Opiliones)

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

A new micro-whip scorpion species from Brazilian Amazonia (Arachnida, Schizomida, Hubbardiidae), with the description of a new synapomorphy for Uropygi

Surazomus uarini n. sp. is described and illustrated based on specimens collected by beating on understory vegetation of Amazonian ""terra firme"" upland rain forests. A new cuticular structure, possibly a gland opening, is described on the female tarsus I and terminal flagellum. A putatively homologous structure is reported from the same body parts in all Undescribed species of Rowlandius Reddell and Cokendolpher 1995; Stenochrus portoricensis Chamberlin 1922; Mastigoproctus maximus (Tarnani 1889), and Thelyphonellus amazonicus (Butler 1872); suggesting it new synapomorphy for the clade Uropygi (i.e., Schizomida + Thelyphonida).

On the utility of female genitalia in the systematics of Curculionidae: examples from Entimini and Hylobiina (Coleoptera : Curculionidae)

The utility of the female genitalia and associated sclerites (tergite and sternite VIII) in the systematics of the Curculionidae is discussed. Examples from the tribe Entimini (Entiminae) and subtribe Hylobiina (Molytinae: Hylobiini) are given. The female characters prove to be informative for establishing the phylogenetic relationships among genera of Entimini. They are essential in determining species groups within the genus Amiticus Pascoe, Hylobiina.

Castoraeschna corbeti sp nov from Floresta Nacional de Carajas, Para state, Brazil (Odonata: Aeshnidae)

Castoraeschna corbeti sp. nov. is described and diagnosed based on four males (holotype: Brazil, Para State, Floresta Nacional cle Carajas [6 degrees 06`13.9 `` S, 50 degrees 08`13.1 `` W, ca 600 m a.s.l.], 28 ix 2007 to be deposited in Museu Nacional, Universidade Federal do Rio de Janeiro, Rio de Janeiro). This species is similar to C. longfieldae and C. coronata but can be distinguished mainly by the absence of medio-dorsal spots on S8; postero-dorsal spots on S8-9 very narrow; cerci external margin almost straight in lateral view, without a distinct angulation between stern and base of lamina; cerci apex blunt. The probable ultimate stadium larva is described based on two individuals, male and female, collected at the type locality. Adults were observed flying along margins of a small shaded second-order stream where the larvae were taken. The surrounding forest is under impact of iron ore extraction and will probably disappear in the next years.

Description of Toca, a new neotropical spider genus (Araneae, Ctenidae, Calocteninae)

Toca new genus is proposed to include two new species: the type species T. bossanova new species from Rio de Janeiro, Brazil, and T samba new species from Parana and Minas Gerais, Brazil. Toca may be related to Caloctenus Keyserling and Gephyroctenus Mello-Leitao, with which it shares the scales on the abdominal dorsum and the epigynum as a single, slightly sclerotized, fold. The genus can be distinguished among the Calocteninae genera by its unique genital structures.

Mating behavior of Sickius longibulbi (Araneae, Theraphosidae, Ischnocolinae), a spider that lacks spermathecae

We describe the mating behavior in the spermatheca-lacking theraphosid species Sickius longibulbi Soares & Camargo 1948. The behavior in captivity of nine pairs of S. longibulbi was videotaped and analyzed. The matting of this species presented an uncommon theraphosid pattern. There is little in the way of overt courtship by the male, the primary behavior seen being the male`s use of legs I and II to touch the female`s first pairs of legs and her chelicerae. Sometimes the male clasped one of the female`s first pairs of legs, bringing her close to him. While the female raised her body, the male clasped her fangs and held her tightly with his legs III wrapped around her prosoma. The male seemed to try to knock the female down, pushing her entire body until she lay on her dorsum. In this phase we observed the male biting the female on the sternum or on the leg joints. When the female fell, the male attempted to position himself at an angle of 90 degrees from the female. These movements appear to demand a lot of energy, particularly because the female is not passive during the mating. Our findings suggest that copulating in this position is, for the male, more successful than adopting other positions because it allows his extremely long palpal bulbs to deposit more sperm in the female oviduct where - since she lacks spermathecae - she retains the sperm. We suggest that the further he reaches into the oviduct, the greater the chance that he will fertilize the female`s eggs.

The identity of Mygale brunnipes CL Koch 1842 (Araneae, Theraphosidae), with a redescription of the species and the description of a new genus

We describe Kochiana new genus to accommodate a small Brazilian theraphosine species described originally as Mygale brunnipes by Koch (1842), resulting in Kochiana brunnipes new combination. Recently, specimens were rediscovered in northeastern Brazilian Atlantic rainforest. A preliminary cladistic analysis using equal weights parsimony and implied weights, was carried out to examine its phylogenetic placement. Kochiana new genus was monophyletic in all trees regardless of weighting scheme or concavity used. There is preliminary evidence for Kochiana new genus monophyly and weak evidence for its placement as sister group of Plesiopelma. Kochiana new genus can be characterized by the presence of a hornshaped spermatheca in females and males with a palpal bulb having prolateral accessory keels and a well developed medial crest on the embolus apex.

Tenacity and silk investment of two orb weavers: considerations about diversification of the Araneoidea

Orbiculariae consists of two major clades: the cribellate Deinopidea and the much more diverse ecribellate Araneoidea. It has been hypothesized that the higher diversity of Araneoidea is a consequence of the superiority of the viscid orb web. However, this explanation seems incomplete: for example, cribellate silk may perform better than viscid silk in some contexts. Here, we consider the hypothesis that the diversification of Araneoidea was facilitated by changes in microhabitat occupation behavior due to the cheaper viscid orb web. In the present work we investigate the idea that the reduction in site tenacity caused by the emergence of the viscid orb web has led to an increase in the exploration of different resources and to a greater diversification of the Arancoidea through the evolutionary time. To test this idea, we evaluated the response of one cribellate orb web spider (Zosis geniculata Olivier 1789, Uloboridae) and one ecribellate orb web spider (Metazygia rogenhoferi Keyserling 1878, Arancidae) to abrupt prey absence. The changes in site tenacity and the day-to-day investment in web silk were evaluated. Spiders with three-dimensional webs tend to exhibit greater site tenacity than spiders making orb webs. Zosis geniculata and M. rogenhoferi show similar site tenacity when prey is ample. When prey is unavailable, the tenacity of the cribellate species increases while the tenacity of the ecribellate remains unchanged, and the silk investment of both species decreases. However, this decrease in silk investment is more extensive in Z. geniculata. These results coincide with the idea that a less costly ecribellate orb web leads to a lower tenacity and suggest that more frequent microhabitat abandonment in a context of insect radiation (Neiptera) leads to more diverse and opportunistic exploration of microhabitats that, in the long term, may be one explanation for the greater Araneoidea diversification.

Resting metabolic rates of two orbweb spiders: A first approach to evolutionary success of ecribellate spiders

Spiders are considered conservative with regard to their resting metabolic rate, presenting the same allometric relation with body mass as the majority of land-arthropods. Nevertheless, web-building is thought to have a great impact on the energetic metabolism, and any modification that affects this complex behavior is expected to have an impact over the daily energetic budget. We analyzed the possibility of the presence of the cribellum having an effect on the allometric relation between resting metabolic rate and body mass for an ecribellate species (Zosis geniculata) and a cribellate one (Metazygia rogenhoferi), and employed a model selection approach to test if these species had the same allometric relationship as other land-arthropods. Our results show that M. rogenhoferi has a higher resting metabolic rate, while Z. geniculata fitted the allometric prediction for land arthropods. This indicates that the absence of the cribellum is associated with a higher resting metabolic rate, thus explaining the higher promptness to activity found for the ecribellate species. If our result proves to be a general rule among spiders, the radiation of Araneoidea could be connected to a more energy-consuming life style. Thus, we briefly outline an alternative model of diversification of Araneoidea that accounts for this possibility. (C) 2011 Elsevier Ltd. All rights reserved.