Contracaecum larvae: morphological and morphometric retrospective analysis, biogeography and zoonotic risk in the amazon / Larva de Contracaecum: análise retrospectiva morfológica e morfométrica, biogeografia e risco zoonótico na amazônia

Abstract In Brazil there are several records of parasitic nematodes of fish with zoonotic potential, especially those belonging to the family Anisakidae. This study considers the morphology, morphometry and prevalence of Contracaecum in Astronotus ocellatus , fish consumed in the Amazon and sold as ornamental and it also performs a retrospective analysis of the diversity of fish with larvae of Contracaecum, in studies carried out in Brazil over a period of 90 years. 40 specimens of A. ocellatus were necropsied, and the nematodes were collected and fixed in 93 parts 70% ethyl alcohol, 5 parts formaldehyde, and 2 parts glacial acetic acid (AFA) for morphological analysis under light microscopy and scanning electron microscopy. Of the 40 fish collected during this work, 27 were parasitized by Contracaecum larvae with a total intensity of 150 larvae. Retrospective analysis of intermediate host diversity for Contracaecum larvae resulted in 16 orders, 49 families, 96 genera, 140 species and a hybrid morphotype. In the retrospective study, half of the fish were from freshwater, with the order Perciformes being the most representative, with 16 families, 30 genera and 37 species. In Brazil, the occurrence of larvae of Contracaecum in fish was reported in 15 of the 26 states, with Rio de Janeiro presenting the most information regarding fish harboring Contracaecum larvae.

Resumo No Brasil existem vários registros de nematóides parasitos de peixes com potencial zoonótico, especialmente aqueles pertencentes à família Anisakidae. Este estudo considera a morfologia, morfometria e prevalência de Contracaecum em Astronotus ocellatus, peixe consumido na Amazônia e vendido como ornamental, e também realiza uma análise retrospectiva da ictiofauna parasitada com larvas de Contracaecum, em estudos realizados no Brasil, durante um período de 90 anos. 40 espécimes de A. ocellatus foram necropsiados, e os nematóides foram coletados e fixados em 93 partes de álcool etílico 70%, 5 partes de formaldeído e 2 partes de ácido acético glacial (AFA) para análise morfológica em microscopia de luz e microscopia eletrônica de varredura. Dos 40 peixes coletados durante este trabalho, 27 estavam parasitados por Contracaecum, com intensidade total de 150 larvas. Análises retrospectivas da diversidade de hospedeiros intermediários para larvas de Contracaecum resultaram em 16 ordens, 49 famílias, 96 gêneros,140 espécies e um morfotipo híbrido. No estudo retrospectivo, metade dos peixes eram de água doce, sendo a ordem Perciformes a mais representativa, com 16 famílias, 30 gêneros e 37 espécies. No Brasil, a ocorrência de larvas de Contracaecum em peixes foi relatada em 15 dos 26 estados, com o Rio de Janeiro apresentando mais informações sobre peixes portadores de larvas de Contracaecum.

Description of fourth instar larva and pupa of Atrichopogon delpontei Cavalieri and Chiossone (Diptera: Ceratopogonidae) from Brazilian Amazonia

ABSTRACT The fourth instar larva and pupa of Atrichopogon delpontei Cavalieri and Chiossone are described for the first time. The immatures were collected from stream margins in the northern Brazilian states Rondônia and Piauí, and subsequently reared to adults. Larvae and pupae are illustrated and photomicrographed. Details on the rearing process and feeding behavior in laboratory, bionomics and notes on habitats are also provided.

Acanthocephala Larvae parasitizing Ameiva ameiva ameiva (Linnaeus, 1758) (Squamata: Teiidae) / Larvas de Acanthocephala parasitando Ameiva ameiva ameiva (Linnaeus, 1758) (Squamata: Teiidae)

Abstract Knowledge concerning the taxonomy and biology of species of Acanthocephala, helminth parasites of the helminth species of the phylum Acanthocephala, parasites of lizards in Brazilian Amazonia, is still insufficient, but reports of Acanthocephala in reptiles are becoming increasingly common in the literature. Cystacanth-stage Acanthocephalan larvae have been found in the visceral peritoneum during necropsy of Ameiva ameiva ameivalizards from the “Osvaldo Rodrigues da Cunha” Herpetology Collection of the Emílio Goeldi Museum, Belém, Pará, Brazil. The aim of this study was to present the morphological study of the Acanthocephala larvae found in A. ameiva ameiva lizard.

Resumo O conhecimento a respeito da taxonomia e da biologia das espécies de Acanthocephala, helmintos parasitos das espécies de lagartos da Amazônia Brasileira ainda é insuficiente, mas o registro do encontro de acantocéfalos em répteis é cada vez mais comum na literatura. Foram encontradas larvas de acantocéfalos em fase cistacanto durante a necropsia, no peritônio visceral, de lagartos Ameiva ameiva ameiva provenientes da Coleção Herpetológica “Osvaldo Rodrigues da Cunha” do Museu Paraense Emílio Goeldi, Belém, Pará. O objetivo é apresentar um estudo morfológico de larvas de Acanthocephala encontradas em A. ameiva ameiva.

THE SUSCEPTIBILITY OF RECENT ISOLATES OF Schistosoma mansoni TO PRAZIQUANTEL

Introduction: Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma and its control is dependent on a single drug, praziquantel (PZQ), but concerns over PZQ resistance have renewed interest in evaluating the in vitro susceptibility of recent isolates of Schistosoma mansoni to PZQ in comparison with well-established strains in the laboratory. Material and methods: The in vitro activity of PZQ (6.5-0.003 µg/mL) was evaluated in terms of mortality, reduced motor activity and ultrastructural alterations against S. mansoni. Results: After 3 h of incubation, PZQ, at 6.5 µg/mL, caused 100% mortality of all adult worms in the three types of recent isolates, while PZQ was inactive at concentrations of 0.08-0.003 µg/mL after 3 h of incubation. The results show that the SLM and Sotave isolates basically presented the same pattern of susceptibility, differing only in the concentration of 6.5 µg/mL, where deaths occurred from the range of 1.5 h in Sotave and just in the 3 h range of SLM. Additionally, this article presents ultrastructural evidence of rapid severe PZQ-induced surface membrane damage in S. mansoni after treatment with the drug, such as disintegration, sloughing, and erosion of the surface. Conclusion: According to these results, PZQ is very effective to induce tegument destruction of recent isolates of S. mansoni.

International cooperation for science and technology development: a way forward for equity in health / Cooperação internacional para o desenvolvimento científico e tecnológico: um caminho para equidade em saúde

Since 1990, international organizations have been increasingly involved in building an international sub-regime for research, development and innovation in health. This article analyzes the priorities of developing countries in health since the 1990s. It is a descriptive and analytical study that investigates the literature and contributions of key informants. Calling for the end of global inequities in the support for science and technology in health, international organizations recommend that developing countries focus their efforts on neglected diseases and operational research, an insufficient agenda for science and technology cooperation to effectively overcome the vulnerabilities between countries.

Desde 1990, intensificou-se o movimento das organizações internacionais na construção do sub-regime internacional de pesquisa, desenvolvimento e inovação em saúde. Este artigo analisa as prioridades dos países em desenvolvimento para essa área desde a década de 1990. Estudo descritivo-analítico, estabelece um diálogo entre a literatura e as contribuições de informantes-chave. Defendendo a superação da iniquidade mundial no fomento científico-tecnológico em saúde, as organizações internacionais recomendam aos países em desenvolvimento focalizar suas ações em doenças negligenciadas e pesquisa operacional - agenda insuficiente para a cooperação científico-tecnológica ser fator de superação de vulnerabilidades entre países.

Alterações ultraestruturais em larvas de Aedes aegypti submetidas ao diterpeno labdano, isolado de Copaifera reticulata (Leguminosae), e à uma fração rica em taninos de Magonia pubescens (Sapindaceae) / Ultrastructural alterations in larvae of Aedes aegypti subject to labdane diterpene isolated from Copaifera reticulata (Leguminosae) and a fraction enriched with tannins of Magonia pubescens (Sapindaceae)

INTRODUÇÃO: Dengue é um importante problema de saúde pública, em vários países, e tem como principal vetor o Aedes aegypti, mosquito mais adaptado às áreas urbanizadas. Apresenta-se, pela primeira vez, as alterações ultraestruturais em larvas de 3º estádio, desse mosquito, causadas pelos larvicidas naturais, um diterpeno labdano, extraído de Copaifera reticulata, e uma fração rica em taninos catéquicos, extraída de Magonia pubescens, evidenciando o mecanismo de ação dessas substâncias. MÉTODOS: Os experimentos foram realizados com larvas de 3º estádio em solução de 0,9ppm, do diterpeno (3-β-acetoxylabdan-8(17)-13-dien-15-óico) e de 3,7ppm, da fração majoritária de tanino catéquico de massa molecular 864Da. Obtiveram-se as substâncias através de fracionamentos cromatográficos sucessivos, identificadas por ressonância magnética nuclear de hidrogênio e espectrometria de massas. As larvas que atingiram estado letárgico foram coletadas e dissecadas e seus tubos digestórios fixados, desidratados, emblocados e polimerizados. Cortes ultrafinos foram feitos e contrastados com acetato de uranila 3 por cento e citrato de chumbo, posteriormente, levados ao microscópio eletrônico. RESULTADOS: As principais alterações ultraestruturais provocadas pelos diterpeno e tanino sobre larvas de Aedes aegypti foram vacuolização citoplasmática, desorganização e degeneração celular, mudança estrutural dos microvilos e deslocamento das células da lâmina basal. CONCLUSÕES: O diterpeno e a fração rica em taninos catéquicos provocaram a morte das larvas de Aedes aegypti através da destruição celular no intestino médio.

INTRODUCTION: Dengue is an important public health problem in many countries and its main vector Aedes aegypti, is the mosquito most adapted to urban areas. For the first time, the mechanism of action of labdane diterpenoid extracted from Copaifera reticulata and the fraction enriched of catechin tannins extracted from Magonia pubescens is demonstrated through ultrastructural alterations of Aedes aegypti larvae. METHODS: Experiments were performed using a 0.9ppm solution of diterpenoid and 3.7ppm of the fraction as the main catechin tannin of molecular mass 846Da. The compounds were obtained by thin layer chromatography and identified by nuclear magnetic resonance of hydrogen and mass spectrometry. Larvae that achieved lethargic state were collected and dissected. Next, they were contrasted with 1 percent uranyl acetate, dehydrated, embedded and polymerized. Ultrathin sections were made, mixed with 3 percent uranyl acetate and lead citrate and placed in an electron microscope. RESULTS: The main ultrastructural alterations caused by the diterpenoid and by tanins in larvae of Aedes aegypti were: cytoplasmic vacuolation, alteration of microvilli, cellular aging, cell disruption and degeneration, formation of secretion vesicles and structural changes in microvilli, irregular nuclei and displacement of cells in the basal lamina. CONCLUSIONS: The fraction containing tannins and the diterpenoid caused the death of Aedes aegypti larvae by cell destruction in the midgut.

Morphology of the larval shell of three oyster species of the genus Crassostrea Sacco, 1897 (Bivalvia: Ostreidae) / Morfologia de conchas larvais de três espécies de ostras do gênero Crassostrea Sacco, 1897 (Bivalvia: Ostreidae)

In this study we describe the morphology of the larval shell of three oyster species of Crassostrea genus. Two species, C. rhizophorae and C. brasiliana, are native to the Brazilian coast, and C. gigas is an introduced species. Samples of laboratory reared larvae, obtained through artificial fertilisation, were collected at intervals during the cultivation process for analysis using Scanning Electron Microscopy (SEM). Prodissoconch morphology was observed in relation to the presence, position, form and number of teeth in the three larval stages: D-shaped larva, umbo larva and pediveliger. Characteristic of D-shaped larvae of C. rhizophorae was the total absence of teeth in the provinculum area while C. brasiliana and C. gigas had two anterior and two posterior teeth in each valve. In the umbo larval phase, the three species had the same number of teeth in each valve: two posterior and two anterior teeth in the right valve and three posterior and three anterior in the left valve. In the pediveliger stage the three species could be differentiated by the number of anterior teeth of the right valve: C. rhizophorae had two teeth, C. brasiliana one tooth and C. gigas three teeth.

Neste estudo, foi descrita a morfologia das conchas larvais de três espécies de ostras do gênero Crassostrea. Duas espécies, C. rhizophorae e C. brasiliana, são nativas da costa brasileira e C. gigas é uma espécie introduzida. Amostras de larvas produzidas em laboratório através de fertilização artificial foram coletadas em intervalos durante o processo de cultivo para análises dos número de dentes nos três estágios larvais: larva D, umbo e pediveliger. As larvas D de C. rhizophorae caracterizaram-se pela total ausência de dentes na área do provinculum, enquanto C. brasiliana e C. gigas apresentaram dois dentes anteriores e dois posteriores em cada valva. Na fase umbo, as três espécies apresentaram o mesmo número de dentes em cada valva: dois dentes anteriores e dois posteriores na valva direita; e três dentes posteriores e três anteriores na valva esquerda. Na fase pediveliger, as três espécies se diferenciaram pelo número de dentes anteriores na valva direita: C. rhizophorae apresentou dois dentes; C. brasiliana, um dente; e C. gigas, três dentes.

Aspectos biológicos e morfologia externa dos imaturos de Memphismoruus stheno (Prittwitz) (Lepidoptera: Nymphalidae) / Biological aspects and external morphology of immature stages of Memphis moruus stheno (Prittwittz) (Lepidoptera: Nymphalidae)

Immature stages of Memphis moruus stheno (Prittwittz) were subject of a behavior, biological and morphological study. The morphological study was carried out through observation by stereoscopic microscopy with camera lucida and micrometric lens attached for illustrations and measurements, respectively; and scanning electron microscopy for ultrastructural analysis. Behavioral data were obtained through direct observation in the field and laboratory. Eggs were laid on the underside of leaves of two Lauraceae species. The first to third instars build frass chains, the fourth and the fifth instars build a conical shelter using a single leaf of the host plant. Before pupation, larvae bend its body ventrally and the pupae are incapable of movement. Descriptions, illustrations and photographs of egg, all five instars and pupa are given. The cephalic morphology and chaetotaxy of first instar were described and illustrated. Results are compared with other Charaxinae immature stages.

Mitochondria in the midgut epithelial cells of sugarcane borer parasitized by Cotesia flavipes (Cameron, 1891) / Mitocôndria nas Células Epiteliais do Intestino Médio da Broca-da-cana Parasitada por Cotesia flavipes

The sugarcane borer Diatraea saccharalis (Lepidoptera: Crambidae) has been controlled by Cotesia flavipes (Hymenoptera: Braconidae); however, very little is known about the effect of the parasitism in the host organs, including the midgut. This work aims to verify mitochondrial alteration in the different midgut epithelial cells of D. saccharalis parasitized by C. flavipes. Midgut fragments (anterior and posterior region) of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. The mitochondria of midgut epithelial cell in the parasitized larvae exhibit morphological alteration, represented by matrix rarefaction and vacuolisation. These mitochondrial alterations are more pronounced in the anterior midgut region during the parasitism process, mainly in the columnar cell.

Diatraea saccharalis (Lepidoptera: Crambidae), broca da cana-de-açúcar, tem sido controlada por Cotesia flavipes (Hymenoptera: Braconidae); pouco se sabe sobre o efeito do parasitismo nos diferentes órgãos do inseto hospedeiro, principalmente no intestino médio. O objetivo desse trabalho foi verificar as alterações mitocondriais das diferentes células epiteliais do intestino médio de larvas de D. saccharalis parasitadas por C. flavipes. Fragmentos do intestino médio (regiões anterior e posterior) de larvas de D. saccharalis não-parasitadas e parasitadas foram processados para microscopia eletrônica de transmissão. As mitocôndrias das células epiteliais do intestino médio de larvas parasitadas exibem alterações, especialmente rarefação e vacuolização da matriz, que foram mais pronunciadas nas células epiteliais da região anterior do intestino médio na vigência do parasitismo, em especial nas células colunares.

Description of the immatures of Linepithema humile Mayr (Hymenoptera: Formicidae)

Linepithema humile Mayr is an ant species originally native to South America that has been spread accidently throughout the globe through international trade. It is a serious urban and crop pest. Despite its economic importance, little is known about the larvae of this species apart from a brief description based on a few specimens. The present investigation is aimed at describing every immature stage of L. humile. Three larval instars were determined through the frequency distribution of the maximum width of head capsules from a sample of 525 larvae. The morphological descriptions were based on 150 eggs, 70 larvae, and 90 pupae examined by light and scanning electron microscopy. Some morphological characteristics reported to be typical of Linepithema Mayr larvae were confirmed - dolichoderoid body shape, presence of dorsal protuberance, sparse simple body hairs, presence of nine pairs of spiracles and dolichoderoid mandibles. We concluded that an earlier published description was based on queen larvae, and that the protuberance is only present in worker larvae. The information provided in this study may aid ant systematics and phylogenetics, as well provide a better understanding of the biology of this species.

Echinococcus granulosus: biological comparison of cattle isolates from endemic regions of Argentina and Spain / Echinococcus granulosus: comparación biológica de aislados de bovinos de regiones endémicas de Argentina y España

In the present study we have compared cattle isolates of Echinococcus granulosus from Argentina and Spain. The aim was to compare and determine if there exist phenotypic and genetic differences within E. granulosus cattle isolates between an endemic area of Spain (where the disease is mainly restricted to a sheep-dog cycle) and an endemic area of Argentina (where cattle are the most abundant intermediate hosts). The Spanish samples were previously identified as G1 genotype. The Argentinean samples were also identified as G1, but some variants were found for the cytochrome c oxidase-1 (CO1) and NADH dehydrogenase-1 (ND1) mitochondrial genes. When comparing the cyst features and the morphology of the larval rostellar hooks in both regions, some differences were found. The morphometric analyses of the larval rostellar hooks showed the existence of two distinct clearly separated groups (one corresponding to the Argentinean samples and the other to the Spanish ones). In conclusion, there are some genetic and phenotypic differences within E. granulosus cattle isolates from Argentina and Spain. Probably these differences, more important from an epidemiological point of view, are related to different steps in the disease control in both countries. Further studies involving other epidemiological, morphometric and molecular data, including other types of livestock, would contribute to clarify and expand the present work.

El objetivo del presente trabajo fue determinar si existen diferencias fenotípicas y genéticas entre los aislados de Echinococcus granulosus de origen bovino provenientes de dos regiones geográficas donde la hidatidosis es endémica, una de España (donde predomina el ciclo perro-oveja) y una de Argentina (donde el bovino es el hospedador intermediario más importante). Las muestras españolas fueron previamente identificadas como pertenecientes al genotipo G1. Las muestras argentinas también correspondían al genotipo G1, pero entre ellas se registraron algunas microvariantes de los genes mitocondriales citocromo c oxidasa-1 (CO1) y NADH deshidrogenasa- 1 (ND1). La comparación de las características de los quistes y de la morfología de los ganchos rostelares del metacestode mostró ciertas diferencias. En conclusión, existen algunas diferencias genéticas y fenotípicas entre los aislados de E. granulosus de Argentina y España. Probablemente estas diferencias, más importantes desde el punto de vista epidemiológico, podrían estar relacionadas con diferentes etapas en los programas de control de la enfermedad en los dos países. Estudios adicionales que involucren datos epidemiológicos, morfométricos y moleculares provenientes de otros tipos de ganado contribuirán a clarificar y ampliar la información aportada por este trabajo.

Pathological and ultrastructural studies on Anisakis simplex rudolphi-1809 infecting Carangoides Bajad with special reference to intestinal maturation in puppies

Thirty five [70%] of 51 Carangoides bajad were naturally infected with Anisakis simplex during the period from September 2007 to January 2008. The fish were collected from eastern south coast of the Red Sea at Hurgada. The morphological and ultrastructures of Anisakis larvae and adults, and the induced lesions in the fish [intermediate host], five puppies [final host] were orally given infected fish. The body of the larvae is gradually tapering towards the anterior part. It is covered by striated ornamentation longitudinally and horizontally, except the anterior region which is smooth. The morphological and ultrastrutural examinations of the anterior body end of larvae showed a prominent boring tooth, 3 pairs of lips inconspicuous and an excretory ventral pore between the rudimentary subventral lips. The anal end showed a distinct mucron and a slit-shaped anus. The pathological studies revealed encapsulated larvae with concentrical fibrous connective tissue infiltrated, with macrophages and lymphocytes on the surface of liver, spleen and peritoneum of the infected fish. The macrophages aggregated together to form the denser part of the capsule, and invaded the adjacent parenchymal tissue. The hepatocytes, under the affected capsule were necrotic and invaded by melanomacrophages

Morphological regional differences of epithelial cells along the midgut in Diatraea saccharalis Fabricius (Lepidoptera: Crambidae) larvae / Diferenças morfológicas regionais das células epiteliais ao longo do intestino médio de larvas de Diatraea saccharalis Fabricius (Lepidoptera: Crambidae)

The sugarcane borer, Diatraea saccharalis Fabricius, is a pest to sugarcane and many other crops. This work aims to characterize morphological variability in the epithelial cells (columnar, goblet and regenerative) along the midgut of D. saccharalis larvae. Fragments of the midgut (anterior, middle and posterior regions) were fixed and processed by light and scanning electron microscopy. There are both cytochemical and ultrastructural differences in the morphology of the epithelial cells, depending on their localization along the midgut. The apical surface of columnar cells shows an increase in both number and size of the apical protrusions from the anterior to the posterior midgut regions. There is an increase in the amount of PAS-positive (Periodic Acid-Schiff Reaction) granules detected in the cytoplasm of both the columnar and regenerative cells, from the anterior to the posterior region. The goblet cell apical surface is narrow in the anterior region, and enlarged in the posterior midgut; the chamber's cytoplasm extrusion are small and thin at the apical cavity surface, being thicker, longer and more numerous at the basal portion of the cavity. Our results suggest that the sugarcane borer midgut has two morphologically different regions, the anterior and the posterior; the middle region is a transitional region.

A broca da cana, Diatraea saccharalis Fabricius, é uma praga da cana-de-açúcar e outras plantações. O objetivo deste trabalho foi caracterizar variações morfológicas nas células epiteliais (colunares, caliciformes e regenerativas) ao longo do intestino médio de larvas de D. saccharalis. Fragmentos do intestino médio (anterior, mediano e posterior) foram fixados e processados para microscopia de luz e eletrônica de varredura. Existem diferenças morfológicas citoquímicas e ultra-estruturais nas células epiteliais, dependendo da sua localização no intestino médio. A superfície apical de algumas células colunares exibe projeções citoplasmáticas que aumentam em número e volume da região anterior para a posterior do intestino médio. Existe aumento dos grânulos PAS-positivos (Reação de Schiff) no citoplasma apical das células colunares e regenerativas, da região anterior para a posterior. A câmara das células caliciformes, na região anterior do intestino médio, mostra seu ápice estreito, enquanto que na posterior essa porção da câmara é alargada; as evaginações citoplasmáticas da câmara são pequenas e finas no ápice, sendo numerosas, longas e mais espessas na porção basal. Os resultados sugerem que o intestino médio da broca da cana apresenta duas regiões morfologicamente distintas, a anterior e posterior; a região mediana é uma região de transição.

Spherites in the midgut epithelial cells of the sugarcane borer parasitized by Cotesia flavipes

Diatraea saccharalis, the main pest of sugarcane, has been controlled by Cotesia flavipes. Very little is known about the effect of parasitism on the host organs, including the midgut. The Lepidoptera midgut epithelium is composed of columnar, goblet, regenerative, and endocrine cells. Spherites have been described in columnar and regenerative cells of several Lepidoptera species, and presented a lot of functional meaning. We identified spherites in the midgut epithelial cells of non-parasitized D. saccharalis larvae analyzed the effect of parasitism on spherite morphology and distribution along the length of the midgut. Midgut fragments of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. All the midgut epithelial cells showed spherites, but they were not preferentially located in a particular part of the cells. Parasitized larvae had more spherites, mainly in the columnar cells, than non-parasitized larvae. This observation was associated with an ionic imbalance within the insect host. Spherites were more abundant in the anterior midgut region than in other regions, which suggests that this region is involved in ion transport by intracellular and/or paracellular route.The morphological variability of spherites in the cells of parasitized larvae was related to the developmental stages of these structures.

Detection of Gnathostoma spinigerum Third-Stage Larvae in Snakeheads Purchased from a Central Part of Myanmar

To examine the infection status of freshwater fish with Gnathostoma spp. larvae in Myanmar, we purchased 15 snakeheads, Channa striatus, from a local market in a suburban area of Naypyidaw, the new capital city. Two larval gnathostomes were collected using an artificial digestion technique, and observed by a light microscope and a scanning electron microscope. The size of an intact larva was 2.65 mm long and 0.32 mm wide. The characteristic morphology of the larvae included the presence of a long esophagus (0.80 mm long), 2 pairs of cervical sacs (0.43 mm long), and a characteristic head bulb with 4 rows of hooklets. The number of hooklets in the 1st, 2nd, 3rd, and 4th row was 45, 48, 50, and 52, respectively. Based on these morphological characters, the larvae were identified as the advanced 3rd-stage larvae of Gnathostoma spinigerum. This is the first report of detection of G. spinigerum 3rd-stage larvae in the central part of Myanmar. Our study suggests that intake of raw meat of snakehead fish in Myanmar may result in human gnathostomiasis.

Morphological method for sexing anopheline larvae.

BACKGROUND & OBJECTIVES: Most of autocidal control of malaria vectors relies on the rearing and release of large numbers of sterile male into a wild population and it would be crucial to separate the males from females before release. This could result in enormous economic benefits in the mass rearing and raise the efficiency of the field operations. The development of genetic sexing of mosquitoes, enabling the release of males only, but impairing the overall fitness of the released insect has been considered greatly. Here we report on a morphological sexing method for the preferential diagnosis and separation of males in late III and IV instar larvae for the mosquitoes Anopheles stephensi Liston and An. culicifacies s.l. (Diptera: Culicidae), the principal vectors of human malaria in Asia and Indian subcontinent. METHODS: Male mosquitoes are identified by their tube like organ at the 9th abdomen segment which originates from segment parallel to the spiracles. Length and width of this organ is measured as 66.66 +/- 9.5 and 14.3 +/- 1.5 microm respectively. The whole length of the organ is 201.63 +/- 23.4 microm. Two fried eggs in the anterior portion of the segment are apparent in males. The length of tube in female is shorter than the male (almost half of the length--37.95 +/- 4.0 microm), its width is slightly stout and wider than the male (16.72 +/- 1.4 microm). Two fried eggs in the anterior portion of the segment are absent. After separation of live male larvae by those characteristics, they were transferred into the trays and emerged adults were identified to ascertain correct identification of sex. RESULTS: All the larvae with male organs developed into male adults with hairy antennae and club shaped palpi, whereas all the female larvae developed into adult females. INTERPRETATION & CONCLUSION: The sex separation at the larval stage will provide a clue for embryonic origin of sex organs, insecticide selection at the larval stage, sex related genes, male sterility and other measures.

Histochemical and ultrastructural evidence of lipid secretion by the silk gland of the sugarcane borer Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae)

The silk gland in Lepidoptera larvae is responsible for the silk production used for shelter or cocoon construction. The secretion of fibroin and sericin by the different silk gland regions are well established. There are few attempts to detect lipid components in the insect silk secretion, although the presence of such element may contribute to the resistance of the shelter to wet environment. This study characterizes the glandular region and detects the presence of lipid components in the secretion of the silk gland of Diatraea saccharalis(Fabricius). The silk gland was submitted to histochemical procedure for lipid detection or conventionally prepared for ultrastructural analyses. Lipid droplets were histochemically detected in both the apical cytoplasm of cell of the anterior region and in the lumen among the microvilli. Ultrastructural analyses of the anterior region showed lipid material, visualized as myelin-like structures within the vesicular Golgi complex and in the apical secretory globules, mixed up with the sericin; similar material was observed into the lumen, adjacent to the microvilli. Lipids were not detected in the cells neither in the lumen of the posterior region. Our results suggest that the silk produced by D. saccharalis has a minor lipid content that is secreted by the anterior region together with the sericin.

A glândula da seda de larvas de Lepidoptera é responsável pela produção da seda usada na construção do casulo ou do abrigo. A secreção de fibroína e sericina pelas diferentes regiões da glândula da seda está bem estabelecida. Existem poucos trabalhos tentando identificar componentes lipídicos na secreção de seda de insetos, embora a presença desse componente contribua para a resistência da seda a ambiente úmidos. Este estudo teve como objetivo identificar a presença de componente lipídico na secreção da glândula da seda de larvas de Diatraea saccharalis (Fabricius), bem como caracterizar a região glandular responsável pela secreção. A glândula da seda foi submetida a procedimentos histoquímicos para detecção de lipídeos e convencionalmente preparada para análise ultra-estrutural. Gotas lipídicas foram histoquimicamente detectadas no citoplasma apical de células da região anterior e no lúmen, entre os microvilos. A análise ultra-estrutural da região anterior mostrou material lipídico, visualizado como figuras do tipo mielina dentro do complexo de Golgi vesicular e nos glóbulos secretores apicais, misturados com sericina; material semelhante foi observado dentro do lúmen, adjacente aos microvilos. Nenhum componente secretor lipídico foi detectado nas células ou no lúmen da região posterior. Os resultados sugerem que a seda produzida pela D. saccharalis tem, pelo menos, um discreto componente lipídico, que é secretado pela região anterior, junto com a sericina.

Reproducción y ultraestructura del huevo y larva de primer estadio de Gyriosomus kingi (Coleoptera: Tenebrionidae) del desierto de Atacama

Reproduction and ultrastructure of egg and first instar larvae of Gyriosomus kingi (Coleoptera: Tenebrionidae) from the desert of Atacama. Gyriosomus is an important Chilean genus of Tenebrionidae, inhabiting the coastal desert of this country. in this environment, the Gyriosomus-species are both eremic and endemic components of the epigean entomofauna. Despite the ecological importance of these species in the Chilean desert, there is a lack of knowledge on the preimaginal stages of them. in this paper we describe the egg and first instar larval morphology of G. kingi Reed, 1873, along with some bionomic aspects of the female. The eggs and larva were obtained from couples kept under laboratory conditions, after being captured during the spring of year 2002, in coastal sites of National Park Llanos de Challe, located in the southern border of the Atacama Desert (28°01’S, 71°03’W). The first third of the chorion presents subhexagonal cells lacking aeropyles. The larvae is oligopod and elongated. its cephalic capsule is strongly chitinized with the prothoracic legs longer and thicker than the rest. According laboratory observations, each egg-laying includes seven to ten eggs each time, with a maximum of six egg-laying per female. The eggs are buried into the soil, covered by a mucilaginous layer which creates a clay-sandy film attached to the chorion. Rev. Biol. Trop. 55 (2): 637-644. Epub 2007 June, 29.

Gyriosomus es un importante género chileno de Tenebrionidae, habitante del desierto costero del país. En este ambiente, las especies de Gyriosomus son un componente erémico y endémico de la entomofauna epígea. A pesar de la importancia ecológica que estas especies tienen en el desierto chileno, poco se conoce respecto de la morfología de los estados preimaginales. En este trabajo se describe la morfología del huevo y de la larva de primer estadio de G. kingi Reed, 1873, y se entregan algunos antecedentes acerca de la bionomía de la hembra. Los huevos y las larvas fueron obtenidos a partir de parejas mantenidas en condiciones de laboratorio. Los adultos fueron capturados durante la primavera del año 2002, en sitios costeros del Parque Nacional Llanos de Challe, localizado en la frontera sur del Desierto de Atacama (28°01’S; 71°03’W). El primer tercio del corión del huevo presenta celdas subhexagonales sin aeropilas. La larva es oligópoda y elongada. Su cápsula cefálica está fuertemente quitinizada y sus patas protorácicas son más largas y más gruesas que las restantes. Según observaciones de laboratorio, cada ovipostura incluye de siete a diez huevos cada vez, con un máximo de seis oviposturas por hembra. Los huevos son enterrados en el suelo, recubiertos con un mucílago que genera una película de arena-arcilla adherida al corión.

The tadpole of Hypsiboas leptolineatus (Braun and Braun, 1977), a species in the Hypsiboas polytaenius clade (Anura; Hylidae)

The larval morphology of Hypsiboas leptolineatus was studied. The tadpole has an ovoid body in lateral view, wider than deep; snout rounded with dorsal reniform nostrils; spiracle sinistral with lateral wall attached to body; anal tube dextral; tail fins convex with acuminate tip; oral disc ventral; labial tooth row formula is 2(2)/3(1); moderately developed beaks with serrated jaw sheaths. These external oral features are compared with those of the known tadpoles in the Hypsiboas polytaenius clade. The oral cavity was studied using an electron microscope. Life history aspects are commented.

A morfologia larval de Hypsiboas leptolineatus foi estudada. O girino possui corpo ovóide em vista lateral, mais largo que alto; focinho redondo com narinas dorsais; espiráculo esquerdo, com parede lateral junto ao corpo; tubo anal destro; nadadeiras convexas, com ponta acuminada; disco oral ventral; fórmula dentária 2(2)/3(1); bico córneo moderadamente desenvolvido serrilhado. Estas características externas são comparadas com as dos outros girinos conhecidos do clado Hypsiboas polytaenius. A cavidade oral foi estudada usando microscopia eletrônica. Aspectos de história natural são comentados.