Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)

Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity.

Systematics of the neotropical fish subfamily Glandulocaudinae (Teleostei: Characiformes: Characidae)

The systematics of the Glandulocaudinae is reviewed in detail and justification for the recognition of the group as a subfamily is discussed. The subfamily Glandulocaudinae consists of three genera: Lophiobrycon with one species plesiomorphic in some anatomical features but some others exclusively derived relative to the species in the other genera; Glandulocauda with two species intermediate in phylogenetic derivation; and Mimagoniates with seven species (one new), all more phylogenetically derived concerning their pheromone producing caudal-fin organs and with other anatomical characters presumably more derived than in the species of the other genera. Glandulocauda melanogenys Eigenmann, 1911, is considered a junior synonym of Hyphessobrycon melanopleurus Ellis, 1911. A replacement name, Glandulocauda caerulea Menezes & Weitzman, is proposed for G. melanopleura Eigenmann, 1911. Gland cells found in the caudal-fin organs of all species are histologically indistinguishable from club cells and probably secrete a pheromone during courtship. The club cells are associated with somewhat modified to highly derived caudal scales forming a pheromone pumping organ in the more derived genera and species. This subfamily is distributed in freshwaters of eastern and southern Brazil, Paraguay, and northeastern Uruguay.

Epigean and subterranean ichthyofauna in Cordisburgo karst area, eastern Brazil

After an ichthyofaunistic survey conducted in May 2007 on surface (epigean) water bodies of Cordisburgo karst area, State of Minas Gerais, 13 species were recorded, mostly characiforms; in addition three non-troglomorphic (normally eyed and pigmented) and one troglomorphic catfish (siluriforms) species were recorded in two caves surveyed at different occasions, totaling 17 fish species for the area. All the nominal species herein reported for Cordisburgo area have been previously reported for the Rio das Velhas basin. None of the species observed in caves were found in epigean habitats and vice-versa. The four cave species are distributed throughout subterranean stream reaches, with individuals at different size/age classes. This, associated to the lack of conspicuous morphological differences in relation to epigean congeners, indicate that Trichomycterus brasiliensis, Gymnotus cf. carapo and Pimelodella cf. vittata are troglophiles (species encompassing individuals able to live and complete their life cycle either in the surface or in the subterranean environment) in the Morena Cave; the latter forms a large population and may be at the beginning of a differentiation process due to isolation in the subterranean habitat, as indicated by a slight reduction in eye size. Topographic isolation may be the cause for the incipient, but unmistakable troglomorphism of the Rhamdiopsis population found in the Salitre Cave, allowing for its classification as troglobite (exclusively subterranean species). The Cordisburgo area is subject to significant anthropic pressure, mainly represented by deforestation for agriculture, cattle raising and timbering. Tourism is an additional important threat for cave communities, calling for urgent protection measures.

Processo de maturação das gônadas de pintado (Pseudoplatystoma corruscans) alimentado com dois níveis proteicos e suplementados com óleo de milho

Investigou-se o efeito da adição de C18:2n6, por meio da inclusão de óleo de milho em dietas com dois níveis de proteína bruta, sobre o processo de maturação de gametas de pintado, Pseudoplatystoma corruscans, mantidos em tanques-rede. Foram avaliados: taxa de sobrevivência, relação peso x comprimento, fator de condição (K) e índice gonadossomático (IGS). O experimento foi realizado entre março de 2004 e fevereiro de 2006, em 12 tanques-rede, distribuídos em seis viveiros-escavados de 600m² e densidade de estocagem de 20 peixes/tanque-rede. Utilizaram-se três tratamentos (T) com duas repetições/viveiro: T1 com 28% de PB; T2 com 28% de PB + 5% óleo de milho e T3 com 40% de PB. O crescimento foi ligeiramente mais alto nos peixes do T3. As taxas de sobrevivência foram acima de 77%. Pode-se inferir que as rações ofertadas não causaram alterações histomorfológicas durante o processo de maturação gonadal dessa espécie. O IGS e o K foram ligeiramente mais altos nos animais alimentados com a ração enriquecida com óleo de milho.

Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7

No effective vaccine or immunotherapy is presently available for patients with the hemolytic uremic syndrome (HUS) induced by Shiga-like toxin (Stx) producedbyenterohaemorragic Escherichia coli (EHEC) strains, such as those belonging to the O157:H7 serotype. In this work we evaluated the performance of Bacillus subtilis strains, a harmless spore former gram-positive bacterium species, as a vaccine vehicle for the expression of Stx2B subunit (Stx2B). A recombinant B. subtilis vaccine strain expressing Stx2B under the control of a stress inducible promoter was delivered to BALB/c mice via oral, nasal or subcutaneous routes using both vegetative cells and spores. Mice immunized with vegetative cells by the oral route developed low but specific anti-Stx2B serum IgG and fecal IgA responses while mice immunized with recombinant spores developed anti-Stx2B responses only after administration via the parenteral route. Nonetheless, serum anti-Stx2B antibodies raised in mice immunized with the recombinant B. subtilis strain did not inhibit the toxic effects of the native toxin, both under in vitro and in vivo conditions, suggesting that either the quantity or the quality of the induced immune response did not support an effective neutralization of Stx2 produced by EHEC strains.

Photodynamic inactivation of four Candida species induced by photogem®

This study evaluated the in vitro susceptibility of C. albicans, C. dubliniensis, C. tropicalis and C. krusei to photodynamic therapy (PDT) induced by Photogem® and light emitting diode (LED). Suspensions of each Candida strain were treated with three photosensitizer (PS) concentrations (10, 25 and 50 mg/L) and exposed to 18.0, 25.5 and 37.5 J/cm² LED light fluences (λ ~ 455 nm). Control suspensions were treated only with PS concentrations, only exposed to the LED light fluences or not exposed to LED light or PS. Sixteen experimental conditions were obtained and each condition was repeated three times. From each sample, serial dilutions were obtained and aliquots were plated on Sabouraud Dextrose Agar. After incubation of plates (37 ºC for 48 hours), colonies were counted (cfu/mL) and the data were statistically analyzed by ANOVA and the Tukey test (α=0.05). Complete killing of C. albicans was observed after 18.0 J/cm² in association with 50 mg/L of PS. C. dubliniensis were inactivated after 18.0 J/cm² using 25 mg/L of PS. The inactivation of C. tropicalis was observed after photosensitization with 25 mg/L and subsequent illumination at 25.5 J/cm². For C. krusei, none of the associations between PS and light resulted in complete killing of this species. PDT proved to be effective for the inactivation of C. albicans, C. dubliniensis and C. tropicalis. In addition, reduction in the viability of C. krusei was achieved with some of the PS and light associations.

Aerobic and anaerobic metabolism for the zebrafish, Danio rerio, reared under normoxic and hypoxic conditions and exposed to acute hypoxia during development

In order to verify the influence of chronic and acute ambient oxygen levels from egg to adult stage of the zebrafish, in vivo oxygen consumption (MO2), critical tensions of oxygen (Pcrit), heart rate (fH) and total body lactate concentration (Lc) were determined for Danio rerio (Hamilton, 1822) raised at 28 °C under normoxic (7.5 mgO2.L-1 or 80 mm.Hg-1) and hypoxic conditions (4.3 mgO2.L-1) and exposed to acute hypoxia during different developmental stages. Our findings confirmed that very early stages do not respond effectively to ambient acute hypoxia. However, after the stage corresponding to the age of 30 days, D. rerio was able to respond to acute hypoxia through effective physiological mechanisms involving aerobic and anaerobic metabolism. Such responses were more efficient for the fishes reared under hypoxia which showed that D. rerio survival capability increased during acclimation to mild hypoxia. Measurements of body mass and length showed that moderate hypoxia did not affect growth significantly until the fish reached the stage of 60 days. Moreover, a growth delay was verified for the hypoxic-reared animals. Also, the D. rerio eggs-to-larvae survival varied from 87.7 to 62.4% in animals reared under normoxia and mild hypoxia, respectively. However, the surviving animals raised under moderated hypoxia showed a better aptitude to regulate aerobic and anaerobic capacities when exposed to acute hypoxia.

Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials.

Ichthyofauna of Rio Jurubatuba, Santos, São Paulo: a high diversity refuge in impacted lands

Ichthyofaunistic surveys in the Atlantic Rainforest have been published in relatively few works, in spite of the major biological importance of this once vast biome which is rapidly vanishing due to disordered human population growth and natural resources overexploitation. The present study aimed to access the fish fauna of a relatively well preserved basin between the cities of Santos and Cubatão (SP), an area highly modified by human activities where recent ichthyofaunistic surveys are still missing. Collections were made during three field trips in Rio Jurubatuba, a medium sized costal river, and Riacho Sabão, one of its main tributaries. A total of 2773 specimens were sampled, representing 25 species from 14 families. Six species were primary marine using the upper reaches of Rio Jurubatuba. Twelve of the 19 freshwater species are endemic of the Atlantic Rainforest and four are present in regional lists of endangered species. Only five species occurred in both Rio Jurubatuba and Riacho Sabão. The most diverse family was Characidae, followed by Poeciliidae, Rivulidae and Heptapteridae. Phalloceros caudimaculatus was the most abundant species, followed by Poecilia vivipara and Geophagus brasiliensis. The study area is considered well preserved and due to its critical location, urges for conservation policies to protect its fish diversity.

Capture fishery in northern Todos os Santos Bay, tropical southwestern Atlantic, Brazil

Information on marine and estuarine capture fishery activity in northern Todos os Santos Bay, northeastern Brazil, based on daily data collected between September 2003 and June 2005 is presented. Small-scale artisanal fishery in this area includes the use of traditional vessels both nonmotorized and motorized for locomotion, being carried out mainly by canoe or on foot, and involves many different kinds of gear, including gillnet, hook and line, seine nets, and traps. A total of 113 taxa were grouped into 77 resources, including 88 fish, 10 crustaceans, and 15 mollusks. Data on nominal catches of fish, crustaceans and mollusks are presented by month and location. A total of 345.2 tonnes of fishery resources were produced (285.4 tonnes of fish, 39.2 tonnes of fresh invertebrates, and 20.6 tonnes of processed invertebrates). Temporal variation in the fish catch was associated with the life cycle of the species or with the hydrographic conditions. The first-sale value of this catch amounted to around US$ 615,000.00, fishes representing 71.3% of it. A table of the average price of each fishery resource is presented. The results produced in this study may be considered a reference for future monitoring programs of fishery resources in the area.

Biologia reprodutiva de Heros efasciatus Heckel, 1840 (Pisces, Cichlidae) na Reserva de Desenvolvimento Sustentável Amanã-AM, visando seu manejo sustentável

Com o principal objetivo de fornecer ferramentas para auxiliar na implementação do manejo sustentável de peixes ornamentais na Reserva de Desenvolvimento Sustentável Amanã, Amazonas, foi realizado o estudo da biologia reprodutiva de Heros efasciatus Heckel, 1840, um ciclídeo com potencial ornamental e com poucos trabalhos sobre a sua biologia e ecologia, apesar de já ser comercializado em algumas regiões amazônicas. Coletas bimestrais foram realizadas de fevereiro de 2006 a janeiro de 2007 em dez igarapés contribuintes do Lago Amanã e Urini, sendo utilizados três aparelhos de pesca (rede de arrasto, rapiché e armadilha tipo matapi) e ainda galhadas artificiais nas amostragens realizadas próximas aos lagos. Foram capturados 140 exemplares de H. efasciatus, sendo 50 fêmeas, 42 machos, e 46 indivíduos cujo sexo não foi identificado devido ao pequeno tamanho. O tipo de crescimento encontrado foi isométrico, sendo que o maior indivíduo observado apresentava 174 mm e o menor 14 mm. Os resultados encontrados auxiliarão na adoção de medidas de manejo, como a determinação de tamanhos mínimos de captura, superiores aos tamanhos médios de maturação (97 mm para as fêmeas) e o estabelecimento de períodos de defeso durante a época de sua reprodução (outubro a janeiro). A pequena abundância de indivíduos da espécie, quando comparada com o total de exemplares capturados (apenas 0,07%) e a baixa fecundidade média, de 2502 ovócitos, indica que se deve trabalhar anualmente apenas com um pequeno número de indivíduos, a fim de garantir a continuidade do estoque.

A new species of Glaphyropoma: the first subterranean copionodontine catfish and the first occurrence of opercular odontodes in the subfamily (Siluriformes: Trichomycteridae)

A new species of the rare copionodontine genus Glaphyropoma is described from subterranean waters in the Diamantina Plateau, Bahia State, central northeastern Brazil. This is the first troglomorphic species in the subfamily Copionodontinae. It is distinguished from all other copionodontines by the presence of opercular odontodes, and further distinguished from its only congener, G. rodriguesi, by the reduction of dark integumentary pigmentation. The new species shares the single synapomorphy previously proposed for Glaphyropoma, the marked narrowing of the first hypobranchial and indirect character evidence also supports its inclusion in the genus. The presence of opercular odontodes in the new species, in combination with a reviewed hypothesis of sister group relationship between Copionodontinae and Trichogeninae, indicate that the absence of opercular odontodes in previously-known copionodontines is secondary, rather than primitive.

Trichomycterus igobi, a new catfish species from the rio Iguaçu drainage: the largest head in Trichomycteridae (Siluriformes: Trichomycteridae)

A new species of Trichomycterus is described for the rio Iguaçu drainage in Southern Brazil. Trichomycterus igobi, new species, is readily distinguishable from all other species currently in the genus by its extremely large head (23.8-26.8 % SL), which is proportionally the largest head in any Trichomycteridae. That characteristic plus the relatively deep body result in a very short-bodied overall aspect, the most extremely such case in the genus Trichomycterus. Other diagnostic features that distinguish the new species from most or all of its congeners include a short caudal peduncle (15.4-19.7 % SL); an almost entirely cartilaginous second hypobranchial (with only vestigial ossification); a mesial expanded palatine ossification; a narrow cleithrum, falciform in shape; and the lack of a proximal posterior concavity on the third ceratobranchial. The new species seems to form a monophyletic group with T. stawiarski and other undescribed species (T. sp. C), also endemic to the rio Iguaçu. As putative synapomorphies, the three species share a rigid spine-like morphology of individual procurrent caudal-fin rays, an extended area of dorsal caudal-fin procurrent rays, and numerous branchiostegal rays (ten or eleven).

Rediscovery of the syntypes of Corydoras meridionalis R. von Ihering, 1911 (Teleostei, Siluriformes, Callichthyidae) and designation of lectotype

Os síntipos de Corydoras meridionalis Rodolpho von Ihering, 1911, tidos como desaparecidos da coleção de peixes do Museu de Zoologia da USP, foram encontrados; os exemplares tinham sido identificados como C. ehrhardti Steindachner, 1910, espécie atualmente considerada sinônimo sênior da primeira. O exame desses sintipos permite confirmar a sinonímia proposta. Dados sobre os síntipos são apresentados e comparados com os dados das descrições originais de C. ehrhardti e C. meridionalis, sendo comentadas as poucas diferenças encontradas entre eles. É designado o lectótipo de C. meridionalis e uma fotografia deste é apresentada.

Peixes da Estação Ecológica Serra Geral do Tocantins, bacias dos Rios Tocantins e São Francisco, com observações sobre as implicações biogeográficas das "águas emendadas" dos Rios Sapão e Galheiros

Um levantamento preliminar da ictiofauna que ocorre na Estação Ecológica Serra Geral de Tocantins, situada no Sudeste do Estado do Tocantins e Noroeste do Estado da Bahia, é apresentado. A Estação Ecológica Serra Geral de Tocantins situa-se no divisor de águas entre as bacias do Rio São Francisco (Rio Sapão) e Rio Tocantins (bacias dos Rios Novo, Balsas e Manuel Alves). A cabeceira comum ou "água emendada" do Rio Sapão e Rio Galheiros, este um afluente do Rio Novo, situa-se no interior da estação e é considerada na literatura como uma possível área de intercâmbio ictiofaunístico entre a bacia do Rio São Francisco e a bacia do Rio Tocantins. Trinta e cinco espécies de peixes foram registradas dentro da Estação Ecológica Serra Geral de Tocantins e em seu entorno imediato, algumas delas desconhecidas da ciência e possivelmente endêmicas da região. Um total de 111 espécies de peixes foi registrado regionalmente (incluindo espécies de peixes registrados nos trechos do Rio Sapão e do Rio Novo/do Sono abaixo da estação). O acará Cichlasoma sanctifranciscense é aqui registrado pela primeira vez na bacia do Rio Tocantins. A ocorrência desta espécie, bem como do lambari Astyanax novae, no Rio Sapão e no Rio Novo/do Sono, são considerados os únicos exemplos inequívocos de transposição natural de espécies de peixes entre as bacias do Rio São Francisco e Tocantins efetuado pelas águas emendadas dos rios Sapão e Galheiros.