Variation in 16S-23S rRNA intergenic spacer regions in Photobacterium damselae: a mosaic-like structure

Phenotypically, Photobacterium damselae subsp. piscicida and P. damselae subsp. damselae are easily distinguished. However, their 16S rRNA gene sequences are identical, and attempts to discriminate these two subspecies by molecular tools are hampered by their high level of DNA-DNA similarity. The 16S-23S rRNA internal transcribed spacers (ITS) were sequenced in two strains of Photobacterium damselae subsp. piscicida and two strains of P. damselae subsp. damselae to determine the level of molecular diversity in this DNA region. A total of 17 different ITS variants, ranging from 803 to 296 bp were found, some of which were subspecies or strain specific. The largest ITS contained four tRNA genes (tDNAs) coding for tRNA(Glu(UUC)), tRNA(LyS(UUU)), tRNA(Val(UAC)), and tRNA(Ala(GGC)). Five amplicons contained tRNA(Glu(UUC)) combined with two additional tRNA genes, including tRNA(Lys(UUU)), tRNA(Val(UAC)), or tRNA(Ala(UGC)). Five amplicons contained tRNA(Ile(GAU)) and tRNA(Ala(UGC)). Two amplicons contained tRNA(Glu(UUC)) and tRNA(Val(UGC)). Two different isoacceptor tRNA(Ala) genes (GGC and UGC anticodons) were found. The five smallest amplicons contained no tRNA genes. The tRNA-gene combinations tRNA(Glu(UUC)) -tRNA(Val(UAC)) -tRNA(Ala(UGC)) and tRNA(Glu(UUC)) -tRNA(Ala(UGC)) have not been previously reported in bacterial ITS regions. The number of copies of the ribosomal operon (rrn) in the P. damselae chromosome ranged from at least 9 to 12. For ITS variants coexisting in two strains of different subspecies or in strains of the same subspecies, nucleotide substitution percentages ranged from 0 to 2%. The main source of variation between ITS variants was due to different combinations of DNA sequence blocks, constituting a mosaic-like structure.

Characterization of the 23S and 5S rRNA genes and 23S-5S intergenic spacer region (ITS-2) of Photobacterium damselae

The 23S ribosomal RNA (rRNA) gene has been sequenced in strains of the fish pathogens Photobacterium damselae subsp. damselae (ATCC 33539) and subsp. piscicida (ATCC 29690), showing that 3 nucleotide positions are clearly different between subspecies. In addition, the 5S rRNA gene plus the intergenic spacer region between the 23S and 5S rRNA genes (ITS-2) were amplified, cloned and sequenced for the 2 reference strains as well as the field isolates RG91 (subsp. damselae) and DI21 (subsp. piscicida). A 100% similarity was found for the consensus 5S rRNA gene sequence in the 2 subspecies, although some microheterogeneity was detected as inter-cistronic variability within the same chromosome. Sequence analysis of the spacer region between the 23S and 5S rRNA genes revealed 2 conserved and 3 variable nucleotide sequence blocks, and 4 different modular organizations were found. The ITS-2 spacer region exhibited both inter-subspecies and inter-cistronic polymorphism, with a mosaic-like structure. The EMBL accession numbers for the 23S, 5S and ITS-2 sequences are: P. damselae subsp. piscicida 5S gene (AJ274379), P. damselae subsp. damselae 23S gene (Y18520), subsp. piscicida 23S gene (Y17901), R damselae subsp. piscicida ITS-2 (AJ250695, AJ250696), P. damselae subsp. damselae ITS-2 (AJ250697, AJ250698).

Solar thermal drying of canarium indicum nuts

Many situations exist in rural areas of developing countries where the help of simple technology can make substantial positive impacts on living conditions, finance, and in this case; sustainability. In the Melanesia region, there are numerous areas identified as needing improvement, including indigenous food preservation which will be addressed with a proposed solar thermal solution utilizing locally available materials as much as possible for low cost local construction. The current knowledge of the drying requirements for the product chosen in this study is quite limited. However, it is believed that solar thermal drying might be feasible for the remote sunny regions as in Melanesia. This paper describes the processes involved in determining the drying parameters of the Canarium indicum nut, and the proposed solar dryer designs that have been considered for the particular environmental conditions and product specifications. Through the selection process, a mixed mode, low-tunnel solar dryer was chosen as the best match to satisfy the different parameters.

Práticas agroecológicas para o cultivo do gergelim (Sesamum indicum L.)

The exceeded use of the natural resources required by the modern agriculture has been caused soil impoverishment, soil salinization and soil compaction. The unreasonable use of chemical fertilizers and pesticides causes chemical imbalances in the plant tissues, nutritional losses, taste chances and human health problems. The monocrops are more vulnerable to the pest and disease attacks. The aim of this work were to indicate the better relative planting time of the cowpea bean for the sesame/cowpea bean intercropping based on the evaluation of agronomic, economic, ecological and physiological parameters and to evaluate the viability of the cow urine and cassava wastewater use as alternative fertilizers on the sesame cultivation. In a field essay, when the sesame/cowpea bean was evaluate, the mainly treatments were the single planting of the two cultures and the multiple cropping, with the cowpea bean being planted on the same time, 7, 14 and 21 days after the sesame. The mainly treatments of the second essay were the combined doses of the cow urine (0 and 50 ml) and cassava wastewater (0, 500 and 1000 mL). In the intercrop evaluation significant differences were found between the treatments for the most of the parameters. It was verified that as the sowing of the cowpea bean was retarded in relation to the sesame sowing, the sesame performance increased when the cowpea bean performance decreased. The treatment 7 days balanced the competition relations that occurred between the two crops in the intercrop. The sesame presented little pronounced responses to the different doses of cassava wastewater. The cow urine affected the sesame growth and the growth rates increased with the increasing of the application of the product. The responses of the sesame growth to the cow urine application must be related both the nutritional richness of the product and the presence of growth stimulating substances. The obtained results indicated that in the sesame/cowpea bean intercropping, when the sesame is planted 7 days after the sesame, there is a higher possibility of the net gains to the farmer. Cow urine and cassava wastewater may be used as additional organic fertilizers

First report of Monticellius indicum Mehra, 1939 (Digenea: Spirorchiidae) infecting Chelonia mydas Linnaeus, 1758 (Testudines: Chelonidae) from Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

EFFECT OF SESAME (SESAMUM-INDICUM L) ON NEST DEVELOPMENT OF ATTA-SEXDENS-RUBROPILOSA FOREL (HYM, FORMICIDAE)

The present study showed a strong deleterious effect on leaf-cutting ant Atta sexdens rubiopilosa nests supplied exclusively with sesame (Sesamum indicum) leaves in the laboratory. Sesame leaves were initially attractive to workers and later became antifeedant. Eucaliptus alba and Hibiscus rosa-sinensis leaves were used as control.

RAPD optimization for genetic studies with Citrulus lanatus and Sesamum indicum genotypes

The Random Amplified Polymorphic DNA (RAPD) technique is powerful for DNA polymorphism determinations and is widely used in research involving different organisms, but it is known that RAPD can be affected by many factors that may result in false positive bands and non-reproducible assays. In this study, we analyzed the effect of several factors such as DNA template, primer and Taq DNA polymerase concentrations to optimize and standardize the RAPD technique for further genetic studies with Citrulus lanattus and Sesamum indicum L. The best combination of DNA, Taq DNA polymerase enzyme and primer concentrations in RAPD amplification procedures for sesame and watermelon genotypes was established.

Estudos de pré-formulação e formulação de Heliotropium indicum (L.) DC (Boraginaceae)

Heliotropium indicum L. (Boraginaceae) é um subarbusto que atinge até 70 cm de altura, é amplamente distribuído geograficamente. A espécie é conhecida como fedegoso na região norte e nordeste. Diante do potencial biológico desta espécie este trabalho objetiva determinar parâmetros de qualidade da droga vegetal até o produto final e avaliar a atividade antimicrobiana de suas folhas, com finalidade de se obter uma formulação fitoterápica semi-sólida. Para isso utilizou-se parâmetros de controle de qualidade físico, químico e físico-químico descritos na Farmacopéia Brasileira e literatura pertinente. A droga vegetal foi classificada como pó grosso, apresentou valores médios de perda por dessecação e cinzas totais de 12,88% e 17,14%, respectivamente. A análise termogravimétrica do pó e do extrato liofilizado mostrou que ambos apresentaram boa estabilidade térmica até 180°C. Os espectros na região do IV mostraram um aumento na intensidade das bandas de absorção do extrato liofilizado, que pode está relacionado à extração dos constituintes químicos da matriz celular. A prospecção química do extrato confirmou a presença de classes de metabólitos secundários já relatados em literatura. A fração clorofórmica sugere a presença de alcalóides pelo teste de precipitação com reagente de Dragendorff. A CCD e a CLAE mostraram uma possível presença de uma mesma substância nas frações alcaloídicas e hexânicas. O extrato bruto de H. indicum L. inibiu o crescimento de Staphylococcus aureus apresentando halos de 12,5 mm±0,707 e 10,5 mm±0,707 para as concentrações de 500 mg/mL e 250 mg/mL, respectivamente. As misturas físicas do extrato com os adjuvantes farmacêuticos, utilizados no desenvolvimento da formulação fitoterápica, não apresentaram incompatibilidade física e não houve modificações significativas no perfil de absorção entre os compostos analisados. A formulação fitoterápica semi-sólida manteve-se estável após sua preparação, após a submissão do gel a força centrípeta e após ação do estresse térmico.

Extratos de sementes de gergelim preto (Sesamum indicum L) obtidos via CO2 supercrítico: isotermas de rendimento global, dados cinéticos e ajustes, ácidos graxos totais, fitosteróis e efeitos neuroprotetores

Na medicina popular brasileira, sementes de gergelim preto (Sesamum indicum L) é um dos mais importantes ingrediente presentes em um chá usado para tratar vítimas de acidente vascular encefálico (AVE). Porém, o isolamento de extratos de gergelim preto para fins medicinais usando a tecnologia do fluido supercrítico não foi realizado. O objetivo deste trabalho foi investigar algumas variáveis de processo da extração com fluido supercrítico em sementes de gergelim preto para gerar extratos aplicáveis a pesquisa do AVE isquêmico focal. Duas isotermas (40 e 60 ºC) foram exploradas, combinadas com pressões de 200-400 bar, em vazão mássica de CO₂ constante de 5,9 x 10^-5 kg/s. Os rendimentos globais foram de 37-53% em base seca. O maior rendimento foi obtido em 60 ºC e 400 bar. A composição de ácidos graxos mostrou uma elevada razão de insaturados/ saturados. A análise de conteúdo de fitosteróis no extrato de maior rendimento revelou maiores quantidades de β-sitosterol + sitostanol, colesterol, campesterol + campestanol + 24-metileno colesterol, Δ-5 avenasterol and estigmasterol, enquanto que menores níveis de Δ-5,24 estigmastadienol, brassicasterol, clerosterol + Δ-5-23 estigmastadienol, Δ-7 avenasterol, eritrodiol and Δ-7 estigmastenol foram observados. As curvas de extração das extrações com fluido supercrítico no menor e maior rendimento mássico (200 e 400 bar a 60 ºC) foram ajustadas pelos modelos de Tan e Liou (1989), Martinez et al. (2003), Esquível et al. (1999), Goto et al. (1993) e Sovová (1994 e 2012). Os modelos de Tan e Liou (1989), Goto et al. (1993) e Sovová (1994 e 2012) apresentaram as melhores valores nominais de somas residuais dos quadrados. Experimentos pilotos sugerem que o extrato obtido via fluido supercrítico de gergelim preto é neuroprotetor em relação a isquemia focal por endotelina-1 no córtex motor de ratos machos adultos, observada a redução da área de infarto isquêmico.

Teste de condutividade elétrica para avaliação da qualidade fisiológica de sementes de gergelim (Sesanun indicum L.)

Two experiments were conducted in order to study the effects of seed number, temperature and time of soaking sesame seeds on the results of the electrical conductivity test for physiological quality of seeds. In the experiment I evaluated the electrical conductivity varying the number of seeds (25, 50 and 100) and temperature (20, 25 and 30° C) in experiment II is different soaking periods (2, 4, 6, 8, 12, 16, 20 and 24 h). The tests were performed with three cultivars (cv. Trebol, cv. Cnpa G4 e cv. Comum). The treatments in factorial 3x3x3 and 3x8 in experiments I and II, respectively. The experimental design was completely randomized, with four replications. Means were compared by Tukey test at 5% probability. The characterization of cultivars held through the germination and vigor tests (physical, physiological and biochemical), which rated the Trebol cultivar seeds with better physiological then the Cnpa G4 and Comum. The electrical conductivity test was conducted with four sub-samples of 25 pure seeds soaked in 75 mL of deionized water at 25° C, to separate the lots from 2 hours of soaking, being feasible to evaluate the quality sesame seeds.

Vacunas inactivas frente a Photobacterium Damselae subsp. Piscicida en dorada (Sparus Aurata): efecto sobre la producción de células IGM positivas y la expresión génetica de moléculas proinflamatorias y MX

Programa de doctorado: Sanidad animal

Malavika et Agnimitra, drama indicum Kalidasae adscriptum;

The Latin translation was never published.