Teste de eficiência de vacinas antiaftosa. II. Relação entre o Índice C em cobaias e o Índice Proteção Camundongos

O exame da eficiência de seis vacinas antiaftosa, realizado com os testes “Indice Proteção Camundongos” e "Índice C” utilizando 15 ou mais cobaias por diluição de vírus na titulação, demonstrou a presença de boa correlação e alta significância entre os resultados das duas provas. O valor do índice Proteção Camundongos foi aproximadamente o dobro do valor do índice C. Assim, o índice Proteção Camundongos pode ser recomendado como teste de eficiência de vacinas antiaftosa. principalmente quando se necessita um grande número de provas, devido a seu baixo custo e facilidade de execução, por usar exclusivamente camundongos.The efficiency of six foot and mouth disease vaccines was examined by the Mouse Protection Index and C Index tests using 15 or more guinea pigs per viral dilution for titration. A good correlation and high significance were obtained between the tests. The value of the Mouse Protection Index was approximately double the C Index value. Thus, in view of its low cost and easy execution due to the exclusive use of mice, the Mouse Protection Index can be recommended for testing the efficiency of foot and mouth disease vaccines, especially when a large number of tests is needed

Teste de eficiência de vacinas antiaftosa. I. Redução da variação do índice C pelo aumento do número de cobaias (Cavia cavia)

Two foot and mouth disease vaccines were submitted to the “C Index” efficiency test with six replicates each using four guinea pigs per viral dilution for titration. The values obtained, when transformed into quality of the vaccine, demonstrated that the same vaccine could be scored as “rejected regular”, “approved good” or “approved very good”, indicating that the random variation in the results may prevent a classification of the immunogen. To determine whether these variations are due to the small number of guinea pigs used, one vaccine was submitted to six replicates using five guinea pigs per viral dilution in the “C Index” test. Analyses of the results using 2 by 2, 3 by 3, 4 by 4 and 5 by 5 arrangements from the data corresponding to all possible combinations when 5, 10, 20, 25 or 30 guinea pigs were used per viral dilution demonstrated that the plus or minus (±) 0.5 log10 variation with 95% confidence limits corresponds to 15 guinea pigs.Duas vacinas antiaftosa foram submetidas a seis repetições cada, à prova de eficiência, “índice C”, usando-se para titulação, quatro cobaias por diluição de vírus. Os valores encontrados, quando transformados em qualidade de vacina, demonstraram que uma mesma vacina poderia ser enquadrada como sendo “reprovada regular”, “aprovada boa” ou “aprovada muito boa”, indicando que a variação dos resultados, dependendo do caso, pode indefinir a classificação do imunógeno. Para verificar se tais variações são devidas ao pequeno número de cobaias, uma vacina foi submetida a seis repetições, usando-se cinco cobaias por diluição do vírus na prova “índice C”. Os arranjos 2 a 2, 3 a 3, 4 a 4, 5 a 5, realizados a partir dos resultados correspondentes a todas as combinações possíveis quando são usadas 5, 10, 15, 20, 25 ou 30 cobaias por diluição viral, demonstraram que a variação de mais ou menos 0,5 logaritmo com 95% de segurança, corresponde a 15 cobaias

Comparison of diode array and electrochemical detection in the C30 reverse phase HPLC analysis of algae carotenoids

Qualitative and quantitative determination of carotenoids pigments can provide valuable information about the organisms in which this important class of compounds is found. In the HPLC analysis of pigments, diode array (DAD), electrochemical (ED) and other kinds of detector may be used. The aim of this work is to develop an HPLC method using a C30 column to identify and quantify sixteen different pigments from algae. A further aim is to compare precision and accuracy obtained by DAD and ED. ED is normally more sensible than DAD. On the other hand, the highest precision and accuracy was obtained with DAD. In conclusion, the method was efficient for quantitative and qualitative analyses of pigments from cyanobacteria and different microalgae classes. Their pigment patterns for several organisms are also discussed.A determinação qualitativa e quantitativa de carotenóides pode fornecer diferentes e importantes informações sobre os organismos que os contêm. Na análise de pigmentos por HPLC diversos detectores podem ser utilizados, como diode array (DAD) e eletroquímico (ED). O presente trabalho tem como objetivo desenvolver um método por HPLC utilizando uma coluna C30 para a identificação e quantificação de dezesseis pigmentos em diferentes classes de algas, além de comparar as respostas obtidas nos detectores DAD e ED por meio da análise dos resultados de precisão e exatidão. Apesar do ED ser geralmente um detector mais sensível que o DAD, os resultados de precisão e exatidão foram mais satisfatórios para o DAD. O método desenvolvido foi eficiente para a análise quantitativa dos pigmentos de cianobactérias e diferentes classes de algas, sendo que o padrão cromatográfico encontrado em cada classe foi discutido.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Milênio-Redoxom

Sesquiterpenes from the essential oil of Laurencia dendroidea (Ceramiales, Rhodophyta): isolation, biological activities and distribution among seaweeds

Two known sesquiterpenes (1R*,2S*,3R*,5S*,8S*,9R*)-2,3,5,9-tetramethyltricyclo[6.3.0.0(1,5)]undecan-2-ol and (1S*,2S*,3S*,5S*,8S*,9S*)-2,3,5,9-tetramethyltricyclo-[6.3.0.0(1,5)]undecan-2-ol were isolated for the first time from the essential oil of the red seaweed Laurencia dendroidea collected in the Brazilian coast. These compounds were not active against eight bacteria strains and the yeast Candida albicans, but showed some antioxidant activity. Both compounds were also found in other seaweed species showing that they are not exclusive taxonomic markers to the genus Laurencia.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Ministério da SaúdeMinistério de Ciência e TecnologiaCNPq - INCT-Redoxom

Antioxidant and antimicrobial properties of 2-(4,5-dihydro-1H-pyrazol-1-yl)-pyrimidine and 1-carboxamidino-1H-pyrazole derivatives

Five previously synthesized 4-trifluoromethyl-2-(5-aryl-3-styryl-1H-pyrazol-1yl)-pyrimidines and six 5-aryl-3-styryl-1-carboxamidino-1H-pyrazole derivatives were screened for their antioxidant proprieties. The antioxidant activities were evaluated by using the DPPH and the HRP/luminol/H2O2 chemiluminescence assay systems and for their antimicrobial activity (MIC). The results were good for those series in some concentration in comparison with the standards.Cinco derivados de 4-trifluorometil-2-(5-aril-3-stiril-1H-pirazol-1il)-pirimidinas e seis 5-aril-3-estiril-1-carboxamidino-1H-pirazois previamente sintetizados foram avaliados de acordo com suas propriedades antioxidantes e antimicrobianas. Estas atividades foram avaliadas por ensaios de DPPH e HRP/luminol/H2O2 quimioluminescência e suas atividades antimicrobianas (CIM). Os resultados foram bons para alguns compostos da série em certas concentrações em comparação direta com padrões.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Growth and microcystin production of a Brazilian Microcystis aeruginosa strain (LTPNA 02) under different nutrient conditions

Cyanobacteria are prokaryotic and photosynthetic organisms, which can produce a wide range of bioactive compounds with different properties; including a variety of toxic compounds, also known as cyanotoxins. In this work, we describe the isolation of seven cyanobacterial strains from two reservoirs in São Paulo State, Brazil. Seven different chemical variants of microcystins (MC-RR, MC-LR, MC-YR, MC-LF, MC-LW, and two demethylated variants, dm-MC-RR and dm-MC-LR) were detected in three of the ten isolated strains. One particular Microcystis aeruginosa strain (LTPNA 02) was chosen to evaluate its growth by cell count, and its toxin production under seven different nutritional regimes. We observed different growth behaviors in the logarithmic growth period for only three experiments (p < 0.05). The total growth analysis identified four experiments as different from the control (p < 0.01). Three microcystin variants (MC-RR, MC-LR and MC-YR) were quantified by liquid chromatography-tandem mass spectrometry. At the experimental end, the toxin content was unchanged when comparing cell growth in ASM-1 (N:P = 1), MLA and BG-11 (N:P = 10) medium. In all other experiments, the lowest microcystin production was observed from cells grown in Bold 3N medium during the exponential growth phase. The highest microcystin content was observed in cultures using BG-11(N:P = 100) medium.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Alfried Krupp von Bohlen und Halbach-StiftungUniversidade de São Paulo Departamento de Análises Clínicas e ToxicológicasSaarland University Institute of Bioanalytical ChemistryUniversidade Federal de São Paulo (UNIFESP) Instituto de Ciências Ambientais, Químicas e FarmacêuticasFriedrich-Schiller University of Jena Institute of NutritionHans Knöll Institute Leibniz Institute for Natural Product Research and Infection BiologyUniversidade de São Paulo Instituto de Química Departamento de BioquímicaUNIFESP, Instituto de Ciências Ambientais, Químicas e FarmacêuticasCNPq: 201609/2012-6SciEL

Effects of heavy metals and light levels on the biosynthesis of carotenoids and fatty acids in the macroalgae Gracilaria tenuistipitata (var. liui Zhang & Xia)

We present here the effect of heavy metals and of different light intensities on the biosynthesis of fatty acids and pigments in the macroalga Gracilaria tenuistipitata (var. liui Zhang & Xia). In order to verify the fatty acid content, gas chromatography with flame ionization detection (GC-FID) was employed. Pigments (major carotenoids and chlorophyl-a) were monitored by liquid chromatography with diode array detection (HPLC-DAD). Cultures of G. tenuistipitata were exposed to cadmium (Cd2+, 200 ppb) and copper (Cu2+, 200 ppb), as well as to different light conditions (low light: 100 µmol.photons.m-2.s-1, or high light: 1000 µmol.photons.m-2.s-1). Cd2+ and Cu2+ increased the saturated and monounsaturated fatty acid content [14:0, 16:0, 18:0, 18:1 (n-7) and 18:1 (n-9)] and all major pigments (violaxanthin, antheraxanthin, lutein, zeaxanthin, chlorophyll-a and &#946;-carotene). Both heavy metals decreased the levels of polyunsaturated fatty acids (PUFA) [18:2 (n-6), 18:3 (n-6), 18:5 (n-4), 20:4 (n-6), 20:5 (n-3), 22:6 (n-3)]. G. tenuistipitata cultures were exposed to high light intensity for five days and no statistically significant differences were observed in the content of fatty acids. On the other hand, the levels of pigments rose markedly for chlorophyll-a and all of the carotenoids studied.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Ministério da SaúdeMinistério de Ciência e TecnologiaCNPq - INCT-Redoxom

Persistência de anticorpos colostrais antiantígeno via ("Virus-Infection - Associated") e anticorpos soroneutralizantes induzidos por infecção natural pelo vírus da febre aftosa em bezerros búfalos indianos (Bubalus bubalis)

Antibody against virus-infection-associated (VIA) antigen and neutralizing serum antibody to foot-and-mouth disease virus were studied in 32 non vaccinated Indian buffalo calves. Antibody levels against VIA antigen were detected by double immunodifusion in 76.5% of the animals at 3 + ½, 30% at 4 + ½ and 6.2% at 5 + ½ months of age. Antibody against VIA antigen was no longer detectable at 7 + ½, 8 + ½ and 9 + ½ months of age. Neutralizing serum antibody levels were studied in 12 of 32 animals. At 3 = ½ months of age neutralizing serum antibody for O1, A24, and C3 virus were found respectively in 83.3%m 83.3% ans 65.6% of samples. At 5 + ½ months of age neutralizing serum antibody was present against the three types only in 20% of the animals and was not detected at 7 + ½ months of age. Twenty one days after the outbreak in cattle when the buffalo calves were 12 + ½ months old antibody against VIA antigen was again detected in 20 of 32 (62.5%) of the animals. Neutralizing serum antibody was found in up to 63.3% of the animals against type A and 16.6% against type O1 and C3.Anticorpos anti-antigeno VIA e anticorpos soroneutralizantes foram estudados em 32 bezerros búfalos indianos não vacinados, a partir de 3 + ½ meses de idade até 1 ano. Esses anticorpos foram detectados até os 5 = ½ meses de idade dos animais, até sofrerem infecção natural do vírus da febre aftosa, o que ocorreu aos 12 + ½ meses de idade. O significados destes achados em relação a susceptibilidade dos bubalinos a essa doença é discutido