Anisakidae Skrjabin & Karokhin, 1945 and Raphidascarididae Hartwich, 1954 nematodes in lutjanidae (pisces: perciformes) from the Brazilian Northeast Coast / Nematodas Anisakidae Skrjabin & Karokhin, 1945 e Raphidascarididae Hartwich, 1954 em Lutjanidae (Pisces: Perciformes) do litoral nordeste brasileiro

Abstract The present study aimed at describing and evaluating the fauna of Anisakidae and Raphidacarididae nematodes of lutjanid fish (snappers) from the Brazilian northeastern coast unloaded at the city of Aracaju, capital of the State of Sergipe. A total of 186 lutjanids of 5 different species were analyzed including 69 Lutjanus analis, 51 L. vivanus , 29 Ocyurus chrysurus, 23 L. synagris, and 14 L. jocu. Nematode specimens found in the viscera of these fish were clarified with lactophenol. Illustrations of each of these specimens were prepared to help identify these samples and compose their taxonomic description and classification. At necropsy, 3,183 nematodes belonging to two families were collected from fish viscera which included the following: Anisakidae: Anisakis, Terranova , Contracaecum, and Goezia; Raphidascarididae: Raphidascaris (Ichthyascaris) and Hysterothylacium . A total of six genera of nematodes were found at postmortem examination of which adult worms of only three genera (Goezia, Raphidascaris and Hysterothylacium) were detected in the fish examined. Larvae of Terranova sp. were found only in the musculature of O. chrysurus . Most of the nematode larvae were present in the mesentery and organs of the fish necropsied. Therefore, the zoonotic potential of these nematodes cannot be ruled out. There was a significant negative correlation between the intensity of parasitism by anisakids and the total body length of O. chrysurus; the intensity of parasitism was greater in smaller and shortef fish. There was a significant positive correlation between the abundance of these parasites and the length of L. synagris; the presence of these parasites increased according to the fish's body length. To the authors' knowledge, these are the first records of Raphidascaris (Ichthyascaris ) sp. and Hysterothylacium sp. in L. vivanus , of Terranova sp. in O. chrysurus and L. jocu, and of Goezia sp. in L. analis. These novel findings add the Brazilian northeast coast and the State of Sergipe in the geographic distribution of these parasites in the country.

Resumo O objetivo deste trabalho é descrever e avaliar a fauna de Anisakidae e Raphidacarididae de peixes lutjanídeos do litoral nordeste do Brasil desembarcados na cidade de Aracaju, capital do estado de Sergipe. Foram analisados 186 lutjanídeos das espécies Lutjanus analis 69, L. vivanus 51, Ocyurus chrysurus 29, L. synagris 23 e L. jocu 14. Exemplares dos nematodas encontrados nas vísceras foram clarificados em lactofenol e ilustrados para auxiliar na identificação e compor a descrição taxonõmica. Foram coletados 3.183 nematodas das vísceras dos peixes, pertencentes a duas famílias: Anisakidae: Anisakis, Terranova, Contracaecum e Goezia; e Raphidascarididae: Raphidascaris (Ichthyascaris) e Hysterothylacium. Totalizando seis gêneros encontrados, apenas nos três últimos gêneros foram encontrados vermes em estágio adulto. Apenas em O. chrysurus foram encontradas larvas de Terranova sp. na musculatura. A maioria das larvas estava restrita ao mesentério e órgãos dos peixes, e em grande intensidade não podendo se descartar o potencial zoonótico. Observou-se uma correlação significativa negativa entre a intensidade anisakídeos e o comprimento total de O. chrysurus, onde a intensidade dos parasitas é maior em peixes de menor comprimento e uma correlação significativa positiva entre a abundância desses mesmos parasitos com o comprimento de L. synagris, onde a presença dos parasitas aumenta conforme o comprimento do peixe. Esse é o primeiro registro de Raphidascaris ( Ichthyascaris) sp. e Hysterothylacium sp. em L. vivanus ; de Terranova sp. em O. chrysurus e L. jocu e Goezia sp. em L. analis, adicionando o litoral nordeste do Brasil e o estado de Sergipe na distribuição geográfica desses parasitos.

Estudo comparativo entre métodos manual e digital no cálculo do ângulo do platô tibial em cães / Comparative study of manual and digital method in the calculation of the tibial plateau angle in dogs

São poucos os dados publicados sobre a aplicabilidade ou a confiabilidade dos métodos digitais para mensuração do ângulo do platô tibial (APT) em comparação à mensuração manual. O objetivo deste estudo foi comparar o APT obtido pelo cálculo manual padrão, em filmes radiográficos, com os valores obtidos por cálculos em imagens digitais, por meio do software do computador. Foram analisadas radiografias digitais do membro pélvico, direito e esquerdo, em perfil verdadeiro, de 20 cães saudáveis, com idade superior a um ano. Três observadores com graus de experiência semelhantes em cálculo do APT realizaram as mensurações em cada radiografia. Primeiramente, foram calculados os ângulos dos platôs tibiais manualmente nas películas radiográficas e, posteriormente, os cálculos foram realizados na imagem digital, por meio do software para medicina veterinária do sistema de raios X digital. Os resultados do presente estudo demonstraram diferenças significativas entre os métodos, com valor das médias do cálculo digital 20,48±3.71 e do cálculo manual 23.90±4.02. Valor de P foi < 0,01 na comparação entre médias dos dois grupos. Na avaliação interobservadores, houve diferença significativa no cálculo manual no perfil direito, o que sugere maior variação no cálculo manual quando comparado ao digital. O cálculo realizado pelo software apresentou menor variação nos valores interobservadores. Acredita-se que a menor variação obtida nos resultados aqui apresentados se dê em razão das facilidades proporcionadas pelo software. Conclui-se que houve diferença entre os valores obtidos por meio dos métodos convencional e digital no cálculo do ângulo do platô tibial, sendo os valores digitais menores entre os observadores, e que o método digital no cálculo do ângulo do platô tibial levou à menor variação quando comparado ao método manual.(AU)

There are few published data on the applicability and reliability of digital methods for measuring the TPA compared with manual measurement. The objective of this study was to compare the TPA obtained by calculating standard manual on conventional radiographs, with the values obtained by calculations in digital images through a computer program. Bilateral digital radiographs of hind limb were analyzed in true profile for 20 healthy dogs older than two years. Three observers with similar levels of experience in calculating the TPA conducted measurements on each radiograph. First we calculated the angle of the tibial plateaus manually in x-ray film and then the calculations were made through the program offered by the computer program in the digital image. The results of this study showed significant differences between the methods, with average value of 20.48±3.71 digital calculating average and 23.90±2.4 in the manual calculation. P value was < 0.01 when comparing averages. No significant difference in manual calculation in right profile were found between interobserver assessments, suggesting greater variation in manual calculation compared to digital. The calculation performed by the software showed less variation in values. The slight variation obtained in our results is believed to be due to the facilities provided by the software. There was a difference between the values obtained by means of conventional and digital methods for calculating the angle of the tibial plateau, and the smaller digital values between the observers and the digital method in the calculation of the tibial plateau angle led to less variation when compared to the manual method.(AU)

Anisakidae Skrjabin & Karokhin, 1945 and Raphidascarididae Hartwich, 1954 nematodes in lutjanidae (pisces: perciformes) from the Brazilian Northeast Coast

Abstract The present study aimed at describing and evaluating the fauna of Anisakidae and Raphidacarididae nematodes of lutjanid fish (snappers) from the Brazilian northeastern coast unloaded at the city of Aracaju, capital of the State of Sergipe. A total of 186 lutjanids of 5 different species were analyzed including 69 Lutjanus analis, 51 L. vivanus , 29 Ocyurus chrysurus, 23 L. synagris, and 14 L. jocu. Nematode specimens found in the viscera of these fish were clarified with lactophenol. Illustrations of each of these specimens were prepared to help identify these samples and compose their taxonomic description and classification. At necropsy, 3,183 nematodes belonging to two families were collected from fish viscera which included the following: Anisakidae: Anisakis, Terranova , Contracaecum, and Goezia; Raphidascarididae: Raphidascaris (Ichthyascaris) and Hysterothylacium . A total of six genera of nematodes were found at postmortem examination of which adult worms of only three genera (Goezia, Raphidascaris and Hysterothylacium) were detected in the fish examined. Larvae of Terranova sp. were found only in the musculature of O. chrysurus . Most of the nematode larvae were present in the mesentery and organs of the fish necropsied. Therefore, the zoonotic potential of these nematodes cannot be ruled out. There was a significant negative correlation between the intensity of parasitism by anisakids and the total body length of O. chrysurus; the intensity of parasitism was greater in smaller and shortef fish. There was a significant positive correlation between the abundance of these parasites and the length of L. synagris; the presence of these parasites increased according to the fishs body length. To the authors knowledge, these are the first records of Raphidascaris (Ichthyascaris ) sp. and Hysterothylacium sp. in L. vivanus , of Terranova sp. in O. chrysurus and L. jocu, and of Goezia sp. in L. analis. These novel findings add the Brazilian northeast coast and the State of Sergipe in the geographic distribution of these parasites in the country.

Resumo O objetivo deste trabalho é descrever e avaliar a fauna de Anisakidae e Raphidacarididae de peixes lutjanídeos do litoral nordeste do Brasil desembarcados na cidade de Aracaju, capital do estado de Sergipe. Foram analisados 186 lutjanídeos das espécies Lutjanus analis 69, L. vivanus 51, Ocyurus chrysurus 29, L. synagris 23 e L. jocu 14. Exemplares dos nematodas encontrados nas vísceras foram clarificados em lactofenol e ilustrados para auxiliar na identificação e compor a descrição taxonõmica. Foram coletados 3.183 nematodas das vísceras dos peixes, pertencentes a duas famílias: Anisakidae: Anisakis, Terranova, Contracaecum e Goezia; e Raphidascarididae: Raphidascaris (Ichthyascaris) e Hysterothylacium. Totalizando seis gêneros encontrados, apenas nos três últimos gêneros foram encontrados vermes em estágio adulto. Apenas em O. chrysurus foram encontradas larvas de Terranova sp. na musculatura. A maioria das larvas estava restrita ao mesentério e órgãos dos peixes, e em grande intensidade não podendo se descartar o potencial zoonótico. Observou-se uma correlação significativa negativa entre a intensidade anisakídeos e o comprimento total de O. chrysurus, onde a intensidade dos parasitas é maior em peixes de menor comprimento e uma correlação significativa positiva entre a abundância desses mesmos parasitos com o comprimento de L. synagris, onde a presença dos parasitas aumenta conforme o comprimento do peixe. Esse é o primeiro registro de Raphidascaris ( Ichthyascaris) sp. e Hysterothylacium sp. em L. vivanus ; de Terranova sp. em O. chrysurus e L. jocu e Goezia sp. em L. analis, adicionando o litoral nordeste do Brasil e o estado de Sergipe na distribuição geográfica desses parasitos.

Impact of insulin concentration and mode of FSH addition on the in vitro survival and development of isolated bovine preantral follicles.

UNLABELLED: The insulin and FSH are two important substances in the folliculogenesis process. Thus, the hypothesis of this experiment is that insulin concentration and the form of FSH addition affect the in vitro survival, growth, and estradiol production after culture of isolated bovine preantral follicles. The effects of insulin concentration (experiment 1) and the influence of both fixed and sequential concentrations of FSH (experiment 2) on the in vitro survival and development of bovine preantral follicles were investigated in this study by IVC for 18 days. In experiment 1, on Day 18 of culture, the addition of insulin at all concentrations promoted follicular survival rates significantly higher than that of the control, with the 10-ng/mL insulin treatment showing values significantly higher than the other treatments. The addition of 5- and 10-ng/mL insulin promoted higher follicular growth than the control and other treatments. In experiment 2, FSH 100 had a higher percentage of follicular viability compared with the control. FSH 100 produced follicle diameters significantly higher than those of the control and FSH seq. TREATMENT: Estradiol levels in the presence of FSH (fixed concentration) were significantly higher than the other treatments. In conclusion, the association of insulin (10 ng/mL) and fixed concentration FSH (100 ng/mL) provides high rates of survival, growth, and estradiol production in bovine preantral follicles.

DNA vaccines against dengue virus based on the ns1 gene: the influence of different signal sequences on the protein expression and its correlation to the immune response elicited in mice.

We analyzed four DNA vaccines based on DENV-2 NS1: pcENS1, encoding the C-terminal from E protein plus the NS1 region; pcENS1ANC, similar to pcENS1 plus the N-terminal sequence from NS2a (ANC); pcTPANS1, coding the t-PA signal sequence fused to NS1; and pcTPANS1ANC, similar to pcTPANS1 plus the ANC sequence. The NS1 was detected in lysates and culture supernatants from pcTPANS1-, pcENS1- and pcENS1ANC-transfected cells and not in cells with pcTPANS1ANC. Only the pcENS1ANC leads the expression of NS1 in plasma membrane, confirming the importance of ANC sequence for targeting NS1 to cell surface. High levels of antibodies recognizing conformational epitopes of NS1 were induced in mice immunized with pcTPANS1 and pcENS1, while only few pcENS1ANC-inoculated animals presented detectable anti-NS1 IgG. Protection against DENV-2 was verified in pcTPANS1- and pcENS1-immunized mice, although the plasmid pcTPANS1 induced slight higher protective immunity. These plasmids seem to activate distinct patterns of the immune system.

Circulating human antibodies against dengue NS1 protein: potential of recombinant D2V-NS1 proteins in diagnostic tests.

The dengue virus (DV) causes one of the most important arthropod-borne human viral diseases throughout the tropical and subtropical countries. However, the morbidity and mortality of DV infections could be reduced with an early hospitalization care and a rapid risk identification of developing the dengue haemorrhagic fever (DHF). The nonstructural glycoprotein 1 (NS1) has been pointed as a reagent for immune-assay diagnostic test optimization. To evaluate this potential, recombinant DV2-NS1 proteins (rNS1) were produced from Escherichia coli (NS1EC) and insect cells (NS1IC) expression. The tests were performed by analysis of a human serum panel reacted against different rNS1 forms. The results demonstrated high correspondence between the DV positive sera and the assay results using native or refolded forms of either NS1IC or NS1EC. Also, the IgG and IgM anti-rNS1 level profiles showed distinct distribution, depending on protein form and disease status. However, the IgM anti-rNS1 reactions did not show sensibility to detect the DV in primary infections. The data obtained from the paired serum samples reactivity comparison suggested a heterogeneous human immune response and absence of correspondence between the IgG and IgM profile levels. Moreover, a patient with negative reference test could be detected by specific IgG anti-rNS1 assays presented here. Therefore, these results sustain the usefulness of dengue nonstructural proteins, in particular the NS1, in diagnostic tests as a complementary reagent.

Different physiological roles of ATP- and PP(i)-dependent phosphofructokinase isoenzymes in the methylotrophic actinomycete Amycolatopsis methanolica.

Cells of the actinomycete Amycolatopsis methanolica grown on glucose possess only a single, exclusively PP(i)-dependent phosphofructokinase (PP(i)-PFK) (A. M. C. R. Alves, G. J. W. Euverink, H. J. Hektor, J. van der Vlag, W. Vrijbloed, D.H.A. Hondmann, J. Visser, and L. Dijkhuizen, J. Bacteriol. 176:6827-6835, 1994). When this methylotrophic bacterium is grown on one-carbon (C(1)) compounds (e.g., methanol), an ATP-dependent phosphofructokinase (ATP-PFK) activity is specifically induced, completely replacing the PP(i)-PFK. The two A. methanolica PFK isoenzymes have very distinct functions, namely, in the metabolism of C(6) and C(1) carbon substrates. This is the first report providing biochemical evidence for the presence and physiological roles of PP(i)-PFK and ATP-PFK isoenzymes in a bacterium. The novel ATP-PFK enzyme was purified to homogeneity and characterized in detail at the biochemical and molecular levels. The A. methanolica ATP-PFK and PP(i)-PFK proteins possess a low level of amino acid sequence similarity (24%), clearly showing that the two proteins are not the result of a gene duplication event. PP(i)-PFK is closely related to other (putative) actinomycete PFK enzymes. Surprisingly, the A. methanolica ATP-PFK is most similar to ATP-PFK from the protozoon Trypanosoma brucei and PP(i)-PFK proteins from the bacteria Borrelia burgdorferi and Treponema pallidum, both spirochetes, very distinct from actinomycetes. The data thus suggest that A. methanolica obtained the ATP-PFK-encoding gene via a lateral gene transfer event.

Consideraciones anatómicas sobre la anastomosis arterial recto-sigmoídea / Anatomical considerations of the recto-sigmoid arterial anastomosis

El conocimiento de la circulación colateral abdominal es importante en las cirugías de la región. Con la finalidad de esclarecer la capacidad de flujos de la anastomosis recto-sigmoídea, su localización y las eventuales variaciones anatómicas, estudiamos las arterias componentes de ella en 43 cadáveres de individuos adultos, brasileños, a los que se inyectó una mezcla de alginato con agua y teñida de tal manera de visualizar su recorrido. Los resultados obtenidos mostraron que la anastomosis recto-sigmoídea está presente en 86.1 por ciento del total de casos; se observó permeable en 70,3 por ciento y se presentó sin permeabilidad en 29,7 por ciento. Estos datos permiten asegurar que esta anastomosis está presente y es funcional en más de la mitad de los casos

New vaccine strategies against enterotoxigenic Escherichia coli. I: DNA vaccines against the CFA/I fimbrial adhesin

Stimulation of the mammalian immune system by administration of plasmid DNA has been shown to be an important approach for vaccine development against several pathogens. In the present study we investigated the use of DNA vaccines to induce immune responses against an enteric bacterial pathogen, enterotoxigenic Escherichia coli (ETEC). Three plasmid vectors encoding colonization factor antigen I (CFA/I), an ETEC fimbrial adhesin, were constructed. Eukaryotic cells transfected with each of these plasmids expressed the heterologous antigen in different compartments: bound to the cytoplasmic membrane (pRECFA), accumulated in the cytoplasm (pPolyCFA) or secreted to the outside medium (pBLCFA). BALB/c mice were intramuscularly (im) inoculated with purified plasmid DNA and the systemic, cellular and secreted CFA/I-specific immune responses were analyzed. The results showed that all three DNA vaccine formulations could elicit CFA/I-specific immune responses. Moreover, cellular location of the plasmid-encoded CFA/I seems to have an important role in the induced immune response. Taken together, these results indicate that DNA vaccines also represent a promising approach against enteric bacterial pathogens

New vaccine strategies against enterotoxigenic Escherichia coli. II: enhanced systemic and secreted antibody responses against the CFA/I fimbriae by priming with DNA and boosting with a live recombinant Salmonella Vaccine

The induction of systemic (IgG) and mucosal (IgA) antibody responses against the colonization factor I antigen (CFA/I) of enterotoxigenic Escherichia coli (ETEC) was evaluated in mice primed with an intramuscularly delivered CFA/I-encoding DNA vaccine followed by two oral immunizations with a live recombinant Salmonella typhimurium vaccine strain expressing the ETEC antigen. The booster effect induced by the oral immunization was detected two weeks and one year after the administration of the DNA vaccine. The DNA-primed/Salmonella-boosted vaccination regime showed a synergistic effect on the induced CFA/I-specific systemic and secreted antibody levels which could not be attained by either immunization strategy alone. These results suggest that the combined use of DNA vaccines and recombinant Salmonella vaccine strains can be a useful immunization strategy against enteric pathogens

Identification of ATP-dependent phosphofructokinase as a regulatory step in the glycolytic pathway of the actinomycete Streptomyces coelicolor A3(2).

The ATP-dependent phosphofructokinase (ATP-PFK) of Streptomyces coelicolor A3(2) was purified to homogeneity (1,600-fold) and characterized (110 kDa, with a single type of subunit of 40 kDa); it is allosterically inhibited by phosphoenolpyruvate. Cloning of the pfk gene of S. coelicolor A3(2) and analysis of the deduced amino acid sequence (343 amino acids; 36,667 Da) revealed high similarities to the PPi-PFK enzyme from Amycolatopsis methanolica (tetramer, nonallosteric; 70%) and to the allosteric ATP-PFK enzymes from other bacteria, e.g., Escherichia coli (tetramer; 37%) and Bacillus stearothermophilus (tetramer, 41%). Further structural and functional analysis of the two actinomycete PFK enzymes should elucidate the features of these proteins that determine substrate specificity (ATP versus PPi) and allosteric (in)sensitivity.

Measurement of lipogenesis in isolated preputial gland cells of the rat and the effect of oestrogen.

Lipogenesis was measured in isolated preputial gland cells of female rats after ovariectomy and after the administration of oestradiol benzoate. Ovariectomy decreased preputial gland cell lipogenesis and also altered the pattern of lipid synthesis, producing a relative decrease in the proportion of polar lipids and an increase in the proportion of 'triglycerides'. Although daily administration of 2 or 10 micrograms oestradiol benzoate for 7 days produced slight increases in preputial gland cell lipogenesis in ovariectomized rats, the effects were not significant. A single injection of 10 micrograms oestradiol benzoate, however, produced significant increases in preputial gland cell lipogenesis of ovariectomized rats at both 2 and 24 h and, moreover, at 24 h the pattern of polar lipid and triglyceride labelling was restored to normal. Prior administration of actinomycin D reduced the lipogenic effect of oestradiol benzoate. Oestradiol benzoate had little or no effect on preputial gland cell lipogenesis in male rats. These results confirm that oestrogen is able to stimulate preputial lipogenesis in female rats. Whether this action of oestrogen is related to its pheromone-producing effect on the preputial glands is not yet known.