Techniques for the In Vitro Production of Queens in Stingless Bees (Apidae, Meliponini)

Considering the ecological importance of stingless bees as caretakers and pollinators of a variety of native plants makes it necessary to improve techniques which increase of colonies' number in order to preserve these species and the biodiversity associated with them. Thus, our aim was to develop a methodology of in vitro production of stingless bee queens by offering a large quantity of food to the larvae. Our methodology consisted of determining the amount of larval food needed for the development of the queens, collecting and storing the larval food, and feeding the food to the larvae in acrylic plates. We found that the total average amount of larval food in a worker bee cell of E varia is approximately 26.70 +/- 3.55 mu L. We observed that after the consumption of extra amounts of food (25, 30, 35 and 40 mu L) the larvae differentiate into queens (n = 98). Therefore, the average total volume of food needed for the differentiation of a young larva of F. varia queen is approximately 61.70 +/- 5.00 mu L. In other words; the larvae destined to become queens eat 2.31 times more food than the ones destined to become workers. We used the species Frieseomelitta varia as a model, however the methodology can be reproduced for all species of stingless bees whose mechanism of caste differentiation depends on the amount of food ingested by the larvae. Our results demonstrate the effectiveness of the in vitro technique developed herein, pointing to the possibility of its use as a tool to assist the production of queens on a large scale. This would allow for the artificial splitting of colonies and contribute to conservation efforts in native bees.

Two new species of Hudsonimyia Roback, 1979 (Diptera: Chironomidae: Tanypodinae) from Neotropical Region unveiled by morphology and DNA barcoding

Two new species of Hudsonimyia Roback, 1979 (Diptera: Chironomidae: Tanypodinae) are described and illustrated as male, pupa and larva. The generic diagnosis of pupa is emended and keys to males, pupae and larvae of known species are provided. The different life stages for one of the described species were associated by DNA barcodes.

Descriptions of Macrolygistopterus subparallelus Pic, 1930 immatures from southeastern Brazil (Coleoptera, Lycidae, Lycinae, Calochromini)

Mature larva and pupa of Macrolygistopterus subparallelus Pic are described and compared to known Calochromini immatures. Larvae were collected alive inside dead trunk in the Atlantic Forest at Estacao Biologica de Boraceia, Salesopolis, Sao Paulo, Brazil. They were maintained in laboratory conditions, and the pupal period was 12 days (one observation). The pupa of this genus is described for the first time. This larva differs from known Calochromini larvae mainly because of its almost cylindrical campodeiform body and longer urogomphi.

NEW RECORDS OF ANOPHELES HOMUNCULUS IN CENTRAL AND SERRA DO MAR BIODIVERSITY CORRIDORS OF THE ATLANTIC FOREST, BRAZIL

Two new records of Anopheles homunculus in the eastern part of the Atlantic Forest are reported. This species was found for the first time in Barra do Ouro district, Maquine municipality, Rio Grande do Sul state, located in the southern limit of the Atlantic Forest. The 2nd new record was in the Serra Bonita Reserve, Camacan municipality, southeast Bahia state. These records extend the geographical distribution of An. homunculus, suggesting that the species may be widely distributed in coastal areas of the Atlantic Forest. It is hypothesized that the disjunct distribution of the species may be caused by inadequate sampling, and also difficulties in species identification based only on female external characteristics. Species identification was based on morphological characters of the male, larva, and pupa, and corroborated by DNA sequence analyses, employing data from both 2nd internal transcribed spacer of nuclear ribosomal DNA and of mitochondrial cytochrome c oxidase subunit I.

Diversification in the use of resources by Idarnes species: bypassing functional constraints in the fig-fig wasp interaction

Mutualisms such as the figfig wasp mutualism are generally exploited by parasites. We demonstrate that amongst nonpollinating fig wasps (NPFWs) parasitic on Ficus citrifolia, a species of Idarnes galls flowers and another species feeds on galls induced by other wasps killing their larvae. The galling wasp inserts its ovipositor through the fig wall into the fig cavity. The ovipositor then follows a sinuous path and is introduced through the stigma and style of the flower. The egg is deposited between the integument and nucellus, in the exact location where the pollinating mutualistic wasp would have laid its egg. Gall induction is a complex process. In contrast, the path followed by the ovipositor of the other species is straightforward: attacking a larva within a developed gall poses different constraints. Shifts in feeding regime have occurred repeatedly in NPFWs. Monitoring traits associated with such repeated evolutionary shifts may help understand underlying functional constraints. (c) 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106, 114122.

The sialotranscriptome of Antricola delacruzi female ticks is compatible with non-hematophagous behavior and an alternative source of food

The hosts for Antricola delacruzi ticks are insectivorous, cave-dwelling bats on which only larvae are found. The mouthparts of nymphal and adult A. delacruzi are compatible with scavenging feeding because the hypostome is small and toothless. How a single blood meal of a larva provides energy for several molts as well as for oviposition by females is not known. Adults of A. delacruzi possibly feed upon an unknown food source in bat guano, a substrate on which nymphal and adult stages are always found. Guano produced by insectivorous bats contains twice the amount of protein and 60 times the amount of iron as beef. In addition, bacteria and chitin-rich fungi proliferate on guano. Comparative data on the transcriptome of the salivary glands of A. delacruzi is nonexistent and would help to understand the physiological adaptations of salivary glands that accompany different sources of food as well as the steps taken by the Acari toward haematophagy, believed to have evolved from scavenging dead animals. Annotation of the transcriptome of salivary glands from female instars of A. delacruzi collected on guano categorized 5.7% of the clusters of expressed genes as putative secreted proteins. They included abundantly expressed TIL-domain-containing proteins (possible anti-microbials), an abundantly expressed protein similar to a serum amyloid found in the sialotranscriptomes of Ornithodoros spp., a savignygrin, a family of mucin/peritrophin/cuticle-like proteins, anti-microbials and an HIV envelope-like glycoprotein also found in soft ticks. When comparing the transcriptome of A. delacruzi with those of blood-feeding female soft and hard ticks some notable differences were observed; they consisted of the following transcripts over- or under-represented or absent in the sialotranscriptome of A. delacruzi that may reflect its source of food: ferritin, mucins with chitin-binding domains and TIL-domain-containing proteins versus lipocalins, basic tail proteins, metalloproteases, glycine-rich proteins and Kunitz protease inhibitors, respectively. (C) 2012 Elsevier Ltd. All rights reserved.

Illustrated key for the identification of the known zoeal stages of brachyuran crabs (Crustacea: Decapoda) from tropical and subtropical Brazil, southwestern Atlantic

Approximately 370 brachyuran species have so far been recorded from the Brazilian coast, 123 of which have had their larval stages fully or partially described. The pictorial guide allows the identification of the first zoea of 110 species. The remaining 13 species with known larval stages are treated to the genus level because of difficulties in the morphological differentiation of closely related species.

Immatures of Dicrepidius Eschscholtz, 1829 and Dipropus Germar, 1839 (Elateridae, Elaterinae, Ampedini: Dicrepidiina)

The larva of Dicrepidius ramicornis (Palisot de Beauvois) and pupa of Dipropus brasilianus (Germar) are described. The larva of D. brasilianus is redescribed. The immatures of both species are illustrated. Comparisons among the known larvae of Dipropus species and between the two genera are presented. A provisional key to Dicrepidiina genera with known larvae is provided.

A review of the genus Corynoneura Winnertz (Diptera: Chironomidae) from the Neotropical region

Currently the genus Corynoneura Winnertz comprises 10 species already cited or described for the Neotropical region. In the present paper 15 new species are described for the Neotropics: 12 new species are described from Brazil, five of them are described as male, female, pupa and larva, Corynoneura canchim sp. n., C. diogo sp. n., C. espraiado sp. n., C. humbertoi sp. n., C. salviniatilis sp. n.; three as male, pupa and larva, C. franciscoi sp. n.; C. longiantenna sp. n. and C. renata sp. n.; the species C. sisbiota sp. n. as male, female and pupa; C. bodoquena sp. n. as male and pupa and C. boraceiasp. n. and C. vidiapodeme sp. n. as males. Corynoneura trondi sp. n. is described from Chile (male, pupa), C. guanacaste sp. n. from Costa Rica (male) and C. zempoala sp. n. from Mexico (male). The knowledge of the distribution of Corynoneura fortispicula, C. sertaodaquina, C. septadentata and C. unicapsulata all species described by Wiedenbrug and Trivinho-Strixino (2011) as well as Corynoneura ferelobata Sublette et Sasa is extended. A larval morphotype is also included. Keys for males, females, pupae and larvae to known species of the Neotropical Corynoneura are given.

Description of immature stages of Phelypera schuppeli (Boheman, 1834) with comments on natural history (Coleoptera: Curculionidae: Hyperinae)

Immatures of the Phelypera schuppeli (Boheman, 1834) (Curculionidae; Hyperinae; Cepurini) are described, illustrated and compared with available descriptions of larvae and pupae of Hyperini. Immatures and adults from midwest (Dourados, Mato Grosso do Sul; Pirenopolis, Goias) and southeast Brazil (Bauru, Sao Paulo) were found on leaves of the host plant, Pachira aquatica Aubl. (Malvaceae, formerly Bombacaceae), a tree used as an ornamental plant in many Brazilian frost-free cities. Larvae of P. schuppeli are exophytic, brightly colored, eruciform and possess abdominal ambulatory ampullae, resembling larvae of Lepidoptera. Mature larvae can spin globular lattice-like cocoons where pupation takes place. Data in the field and under laboratory conditions confirmed previously published biological observations on P. schuppeli. Additional information about defensive behaviors, process of cocoon construction and natural enemies, such as the larval predator Supputius cinticeps (Stal, 1860) (Hemiptera: Pentatomidae) and the prepupal and pupal parasitoid Jaliscoa nudipennis Boucek, 1993 (Hymenoptera: Pteromalidae), are reported.

The influence of captive breeding on the fatty acid profiles of Salminus hilarii (Characiformes: Characidae) eggs and larvae

In addition to the strong influence of the broodstock diet on the development and survival of offspring, domestication may also interfere with the larval life success. We obtained eggs from wild and domesticated Salminus hilarii females and domesticated males. Wild females were caught in the Tiete River and tributaries, and the domesticated females were born three years before the beginning of the experiment in the Ponte Nova Fish Farm. Animals from both groups were fed with the same feed to exclude feed variables. The eggs and larvae were sampled at 0, 8, 16, and 28 h after spawning (HAS), with the last sampling (28 HAS) coinciding with hatching time. After hatching, samplings proceeded at 32, 48, 66, and 96 HAS, with the last sampling (96 HAS) corresponding to the end of yolk sac consumption. Finally, the last experimental period was during the larvae exogenous feeding phase, at 102, 118, 166, and 214 HAS. Our data revealed that domestication of S. hilarii females influenced fatty acid (FA) metabolism during embryo and larva development. However, the structure of membrane phospholipid FA remained mostly stable, with changes principally in the neutral fraction. When the external conditions, mainly water and feed quality, remained constant, domestication of S. hilarii females did not significantly affect the structural FA composition but influenced the selectivity of consumption and/or storage of specific FA.

Molting dynamics and juvenile hormone titer profiles in the nymphal stages of a lower termite, Cryptotermes secundus (Kalotermitidae) - signatures of developmental plasticity

Termites are social cockroaches and this sociality is founded on a high plasticity during development. Three molting types (progressive, stationary and regressive molts) are fundamental to achieve plasticity during alate/sexual development, and they make termites a major challenge to any model on endocrine regulation in insect development. As the endocrine signatures underpinning this plasticity are barely understood, we studied the developmental dynamics and their underlying juvenile hormone OH) titers in a wood-dwelling termite. Cryptotermes secundus, which is characterized by an ancestral life style of living in dead wood and individuals being totipotent in development. The following general pattern elements could be identified during winged sexual development (i) regressive molts were accompanied by longer intermolt periods than other molting types, (ii) JH titers decreased gradually during the developmental transition from larva (immatures without wing buds), to nymph (immatures with wing buds), to winged adult, (iii) in all nymphal stages, the JH titer rose before the next molt and dropped thereafter within the first week, (iv) considerable variation in JH titers occurred in the midphase of the molting cycle of the 2nd and 3rd nymphal instar, inferring that this variation may reflect the underlying endocrine signature of each of the three molting types, (v) the 4th nymphal instar, the shortest of all, seems to be a switch point in development, as nymphs in this stage mainly developed progressively. When comparing these patterns with endocrine signatures seen in cockroaches, the developmental program of Cryprotermes can be interpreted as a co-option and repetitive use of hormonal dynamics of the post dorsal-closure phase of cockroach embryonic development. (C) 2012 Elsevier Ltd. All tights reserved.

Movement of Heterorhabditis amazonensis and Steinernema arenarium in search of corn fall armyworm larvae in artificial conditions

Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) is considered to be the main pest of maize crops in Brazil. Entomopathogenic nematodes (EPN) may be used to control this pest and exhibit different, unique abilities to search for their hosts. The movement of EPN in relation to S. frugiperda was evaluated. To test for horizontal movement, a styrofoam enclosure filled with sand was divided into segments, nematodes were placed at the entrance to the enclosure and a larva was placed at the end of each division. The same approach was used to evaluate vertical movement; however, PVC pipes were used in this case. In general, the mortality was inversely proportional to the initial distance between host and nematodes. In the vertical displacement test, both nematodes were able to kill the larvae up to a distance of 25 cm. Therefore, the infective juveniles of H. amazonensis and S. arenarium can search out, infect and kill larvae of S. frugiperda at distances of up to 60 cm and 25 cm of horizontal and vertical displacement, respectively.

Cardiac markers: profile in rats experimentally infected with Toxocara canis

The aim of this study was to evaluate the profile of the enzymes creatine kinase (CK), creatine kinase MB (CK-MB) and lactate dehydrogenase (LDH) in Wistar rats infected with 250 (GI, n = 24) or 1000 (GII, n = 24) Toxocara canis eggs. Animals were evaluated on days 7, 15, 30, 60, 120 and 180 post-infection (DPI). Only the GI rats showed an increase in CK and CK-MB, at 15 and 30 DPI, respectively. Anti-Toxocara spp. antibodies were detected by ELISA in infected animals. Despite of the presence of eosinophilic infiltrate in the heart of three infected animals, none larva was recovered from the organ neither by acid digestion nor by Baermann procedure. Eosinophilia was observed in both groups but there was no significant difference in the eosinophil counts between GI and GII (p = 0.2239). It is possible to consider that cardiac lesion is an eventual finding in murine model for toxocariasis.

Human toxocariasis: diagnosis, worldwide seroprevalences and clinical expression of the systemic and ocular forms.

Although human toxocariasis ranks among the most common zoonotic infections worldwide, it remains relatively unknown to the public. The causal agents are the nematode parasites Toxocara canis and T. cati, whose definitive hosts are dogs and cats, respectively. When embryonated eggs are accidentally ingested by humans, larvae hatch in the small intestine, penetrate the intestinal wall and migrate, via the bloodstream, to the liver, lungs, muscles, eye and central nervous system. Although most human infections are asymptomatic, two well-defined clinical syndromes are classically recognised: visceral larva migrans (a systemic disease caused by larval migration through major organs) and ocular larva migrans (a disease limited to the eyes and optic nerves). Two less-severe syndromes have recently been described, one mainly in children (covert toxocariasis) and the other mainly in adults (common toxocariasis). Here, the current laboratory diagnosis, epidemiology and main clinical features of both the systemic and ocular forms of human toxocariasis are reviewed. New developments in serological diagnosis are described, the available seroprevalence data are analysed, and the results of relevant clinical studies that have been published over the last decade are explored, to provide an updated overview of this neglected but highly prevalent human infection.