The experimentally pathogenesis of Aeromonas hydrophila in Astacus leptodactylus with emphasis on temperature in bacterial pathogenesis

In this study, four hundred freshwater Crayfish (A. leptodactylus) with average weight of 25-40g were purchased from Aras dam reservoirs in west Azerbayjan province and transported to Iranian Artemia Research Center of Uromya province in September 2010. (One hundred crayfish extra purchased for probably mortality). Before implement of experiment the Crayfish were acclimated for ten days. These experiments was designed in four group treatments (Number, 1,2,3,4) and one control group (Number 5) in triplicate with 20 Crayfish in each repetition prepared of glass aquarium with size (50x40x30cm). Many of infected Crayfish were used for isolation of bacteria. Haemolymph sample had been gathered from infected Crayfish with cutting their antennules and transferred to TSA medium (tryptic soy agar) and then A. hydrophila were determined in order to biochemical test. The treatments and repetitions has exposed to A. hydrophila. The concentrations of the bacteria in 4 treatments were respectively 3 x 108 (T=10-15°C), 3x106 (T=25°C), 3 x 106 and 3 x 104 Cfu mL-1 (T=10- i5oc) (4, 2, 3 and 1) that were prepared in individual containers for exposure of treatments. The control (5) prepared without any bacteria and disinfected by oxytetracycline antibiotic with concentration 100 ppm for 24 hours. The hemolymph samples were withdrawn from abdominal second segments of Crayfish for measuring of THC and TPC in interval hours (2, 6, 12, 24, 48, 96, 144, 240 and 336). For histopatological studies the crayfish samples fixed in Davidson fixative. The results indicated that interval 2 hours after experiment the difference of THC value between treatment 4 with control and treatments 1,2, and 3 was significant (P< 0.05). After 48 hours of experiment the difference of THC value between control group with treatment 1 ,2 and 3 was significant (P< 0.05). The interval 336 hours after experiment also the difference of THC value between treatment 2 with treatments 1, 3 and 4 was significant (P< 0.05). The finding of TPP value showed that the last time after challenge (336 h) there was significant difference between treatment 2 with treatment 4 and control group (P< 0.05). In histopathology studying, in hepatopancreas observed hemocyte aggregated and necrosis withof peknosis nucleus that with increased concentration of bacteria and temperature, The value of hemocyte has increased. Gill revealed necrosis and cell death especially with increased concentration of bacteria and temperature. In lower concentration of bacteria in heart no difference observed, but with increased concentration of bacteria (3 x 108) the low aggregation of hemocyte observed in heart. In treatment 3 x 106 with high temperature also distributed of high hemocyte in heart was observed. In digestive system didn't appear any difference in treatments land 3 but in concentration of 3 x 108 Cfu m1-1 and 3 x 106 (T=25°C) in digestive system was revealed the low aggregation of hemocyte.

Biomarkers of Nitrosative Stress: Development and validation of a new analytical method for 3-Nitrotyrosine quantification

Background: The nitration of tyrosine residues in proteins is associated with nitrosative stress, resulting in the formation of 3-nitrotyrosine (3-NT). 3-NT levels in biological samples have been associated with numerous physiological and pathological conditions. For this reason, several attempts have been made in order to develop methods that accurately quantify 3-NT in biological samples. Regarding chromatographic methods, they seem to be very accurate, showing very good sensibility and specificity. However, accurate quantification of this molecule, which is present at very low concentrations both at physiological and pathological states, is always a complex task and a target of intense research. Objectives: We aimed to develop a simple, rapid, low-cost and sensitive 3-NT quantification method for use in medical laboratories as an additional tool for diagnosis and/or treatment monitoring of a wide range of pathologies. We also aimed to evaluate the performance of the HPLC-based method developed here in a wide range of biological matrices. Material and methods: All experiments were performed on a Hitachi LaChrom Elite® HPLC system and separation was carried out using a Lichrocart® 250-4 Lichrospher 100 RP-18 (5μm) column. The method was further validated according to ICH guidelines. The biological matrices tested were serum, whole blood, urine, B16 F-10 melanoma cell line, growth medium conditioned with the same cell line, bacterial and yeast suspensions. Results: From all the protocols tested, the best results were obtained using 0.5% CH3COOH:MeOH:H2O (15:15:70) as the mobile phase, with detection at wavelengths 215, 276 and 356 nm, at 25ºC, and using a flow rate of 1 mL/min. By using this protocol, it was possible to obtain a linear calibration curve (correlation coefficient = 1), limits of detection and quantification in the order of ng/mL, and a short analysis time (<15 minutes per sample). Additionally, the developed protocol allowed the successful detection and quantification of 3-NT in all biological matrices tested, with detection at 356 nm. Conclusion: The method described in this study, which was successfully developed and validated for 3-NT quantification, is simple, cheap and fast, rendering it suitable for analysis in a wide range of biological matrices.

Evaluación y producción masiva de hongos entomopatógenos para el manejo del psilido de la papa (Bactericera cockerelli Sulc) en cultivos de tomate (Solanum lycopersicum L.)y chile (Capsicum annuum L.) en invernadero

En la búsqueda de nuevas estrategias para el manejo de la Paratrioza, Bactericera cockerelli, se ha incluido recientemente el uso de hongos entomopatógenos, principalmente debido a su modo de acción. En este trabajo se evaluaron cuatro cepas de hongos entomopatógenos, dos pertencientes a Beauveria bassiana (BB09 y BB42) y dos a Metarhizium anisopliae (MA25 y MA28). Para las cuatro cepas se valoraron cuatro medios de cultivo (Agar Agua, Papa Dextrosa Agar, Papa Dextrosa Agar + 5% Levadura y Papa Dextrosa Agar + 5% Sacarosa) y tres temperaturas diferentes (20, 25 y 30°C); de igual manera, las cuatro cepas se sujetaron a prueba en tres diferentes medios de producción masiva (arroz, bagazo de caña y bagazo de uva). Para estas tres pruebas se evaluó crecimiento de micelio, viabilidad y esporulación. De manera conjunta se observó la adhesión de conidias de cada una de las cepas a ninfas de B. cockerelli bajo condiciones de laboratorio. Para concluir las evaluaciones, se realizaron dos experimentos en invernaderos para valorar las mortalidades generadas en dos cultivos diferentes (chile y tomate) a dos diferentes concentraciones (1x107conidias por mL y 1x108conidias por mL). De las evaluaciones se concluyó que para éstas cepas el medio de cultivo más adecuado es aquél enriquecido con levadura y que dichas cepas se desarrollan mejor a 25°C, a excepción de las cepas de B. bassiana, las cuales generan mayor cantidad de esporas a 30°C; en cuanto a la reproducción masiva, la búsqueda de sustratos alternativos debe continuar, ya que el bagazo de uva y el de caña no fueron capaces de dar las condiciones adecuadas para generar la misma cantidad de esporas que en el medio usual, el arroz. Finalmente, utilizar hongos entomopatógenos para el manejo de B. cockerelli bajo condiciones de invernadero promete ser una estrategia confiable, ya que dependiendo de la concentración y la cepa utilizada, pueden lograrse niveles de control que van desde un 50 a un 82%.

Óleos essenciais e extratos vegetais no controle de fitopatógenos de grãos de soja.

A utilização de sementes sadias é um importante método para diminuir a ocorrência de doenças desde o inicio de seu cultivo. Uma das alternativas ao uso de agrotóxicos é a utilização de óleos essenciais e extratos de plantas que possuam propriedades tóxicas aos fitopatógenos. O objetivo deste trabalho foi avaliar o efeito de óleos essenciais e extratos hidroalcoólicos de plantas medicinais na sanidade e germinação de sementes de soja. Os óleos essenciais e os extratos hidroalcoólicos foram diluídos em água destilada, na concentração de 2 mg/mL para os extratos e 20% para os óleos essenciais. Em seguida, as sementes foram pulverizadas com os tratamentos e secas a sombra durante 4 horas em temperatura ambiente (25ºC). Os tratamentos utilizados foram: T1=testemunha (água destilada); T2 = óleo essencial de gengibre; T3 = óleo essencial de limão Taiti; T4 = extrato de pariparoba e T5 = extrato de penicilina. Os testes de germinação e sanidade foram realizados segundo as Regras para Análises de Sementes (RAS). Os resultados obtidos no teste de germinação demonstraram que o percentual de germinação inicial das sementes não tratadas (4%) encontrava-se muito abaixo dos níveis recomendados pelas RAS (80%), não tendo como avaliar a interferência dos tratamentos na germinação. Em função deste resultado, foi realizado o teste de sanidade visando o tratamento de grãos. O óleo essencial de gengibre apresentou uma redução da incidência de 75% de Cladosporium sp; Rhizopus sp.; Fusarium spp. quando comparado à testemunha. Pode-se concluir que o óleo essencial de gengibre na concentração de 20% apresenta atividade positiva no controle de Cladosporium sp, Rhizopus sp e Fusarium spp em grãos de soja.

Estudo da sinterização de cavacos de aço ferritico

This research studies the sintering of ferritic steel chips from the machining process. Were sintered metal powder obtained from machining process chips for face milling of a ferritic steel. The chip was produced by machining and characterized by SEM and EDS, and underwent a process of high energy mill powder characterized also by SEM and EDS. Were constructed three types of matrixes for uniaxial compression (relation l / d greater than 2.5). The differences in the design of the matrixes were essentially in the direction of load application, which for cylindrical case axial direction, while for the rectangular arrays, the longer side. Two samples were compressed with different geometries, a cylindrical and rectangular with the same compaction pressure of 700 MPa. The samples were sintered in a vacuum resistive furnace, heating rate 20 °C / min., isotherm 1300 °C for 60 minutes, and cooling rate of 25 °C / min to room temperature. The starting material of the rectangular sample was further annealed up to temperature of 800 ° C for 30 min. Sintered samples were characterized by scanning electron microscopy, optical microscopy and EDS. The sample compressed in the cylindrical matrix did not show a regular density reflecting in the sintered microstructure revealed by the irregular geometry of the pores, characterizing that the sintering was not complete, reaching only the second phase. As for the specimen compacted in the rectangular array, the analysis performed by scanning electron microscopy, optical microscopy and EDS indicate a good densification, and homogeneous microstructure in their full extent. Additionally, the EDS analyzes indicate no significant changes in chemical composition in the process steps. Therefore, it is concluded that recycling of chips, from the processed ferritic steel is feasible by the powder metallurgy. It makes possible rationalize raw material and energy by manufacture of known properties components from chips generated by the machining process, being benefits to the environment

¿Cómo combatir el estrés calórico (II)?

En una primera parte de este artículo, publicado en Entorno Ganadero No. 66, el autor hace referencia a las condiciones ambientales reinantes en un alojamiento de vacuno lechero, las cuales son la mayor importancia para el confort y bienestar de los animales allí alojados, además de explicar que es una condición necesaria (aunque no suficiente) para que éstos puedan expresar todo su potencial productivo. También explicó que cuando las vacas están en unas condiciones de alojamiento óptimas aumenta su resistencia a las enfermedades, es decir, a la primera causa de NO BIENESTAR. Lo anterior debido a que el sistema inmunitario del animal se deprime cuando estas “necesidades ambientales” no están correctamente satisfechas. Ya entrado en materia de este trabajo se refirió fundamentalmente al factor temperatura, cuyos elevados valores van a causar lo que se conoce como “estrés térmico” o “estrés calórico”. Detallando que no obstante, otros factores ambientales como la humedad o la velocidad del aire pueden aliviar o agravar este estrés. Dentro del factor de Temperatura, detalló puntos como: Mecanismos de producción de calor; Mecanismos de eliminación de calor y Temperatura ambiental óptima. En el punto de Evaluación del Estrés Calórico, explicó entre otras cosas, que las vacas lecheras prefieren temperaturas entre 0 y 24ºC, pudiendo mantener su producción incluso a temperaturas de -10ºC. Sin embargo, las vacas empiezan a experimentar estrés por calor a una temperatura de 25ºC, con niveles normales de humedad relativa. En esta segunda entrega, se analizarán puntos como la Consecuencia del Estrés Calórico, los métodos para reducirlo, y se detallará el concepto de refrigeración.

Efeito das variáveis laboratoriais protéticas na adesão da porcelana com ligas de níquel-cromo

The metalceramic crowns are usually used in dentistry because they provide a resistant structure due to its metallic base and its aesthetics from the porcelain that recovers this structure. To manufacture these crowns, a series of stages should be accomplished in the prosthetic laboratories, and many variables can influence its success. Changes in these variables cause alterations in the metallic alloy and in the porcelain, so, as consequence, in the adhesion between them. The composition of the metal alloy can be modified by recasting alloys, a common practice in some prosthetic laboratories. The aim of this paper is to make a systematic study investigating metalceramic crowns as well as analyzing the effect of recasting Ni-Cr alloys. Another variable which can influence the mechanism of metalceramic union is the temperature used in firing porcelain procedure. Each porcelain has to be fired in a fixed temperature which is determined by the manufacturer and its change can cause serious damages. This research simulate situations that may occur on laboratory procedures and observe their consequences in the quality of the metalceramic union. A scanning eletron microscopy and an optic microscopy were accomplish to analyse the metal-ceramic interface. No differences have been found when remelting alloys were used. The microhardness were similar in Ni-Cr alloys casted once, twice and three times. A wettability test was accomplished using a software developed at the Laboratório de Processamento de Materiais por Plasma, on the Universidade Federal do Rio Grande do Norte. No differences were found in the contact angle between the solid surface (metallic substratum) and the tangencial plane to the liquid surface (opaque). To analyse if the temperature of porcelain firing procedure could influence the contact area between metal and porcelain, a variation in its final temperature was achieve from 980° to 955°C. Once more, no differences have been found

Citotoxicidade de curcumina presente em nanoparticulas de quitosana/sultato de condroitina contendo lecitina

Dissertação (mestrado)–Universidade de Brasília, Universidade UnB de Planaltina, Programa de Pós-Graduação em Ciência de Materiais, 2015.

Análise da interação entre moléculas desenhadas a partir do ácido anacárdico e a proteína PPAR usando ferramentas in silico

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Molecular, 2016.

Use of Bioscreen C for growth of Mucor hiemalis in different carbon and nitrogen sources

O sistema automatizado Bioscreen C foi utilizado para o crescimento de quatro linhagens de Mucor hiemalis, isoladas do solo da Estação Ecológica de Juréia-Itatins (EEJI), estado de São Paulo, em meios líquidos com uma única fonte de carbono (2%) ou de nitrogênio (1%), pH 5,0, a 25ºC, e agitação de 150rpm por 5 dias. O meio com somente uma única fonte de nitrogênio foi adicionado com 2% de glicose. As leituras de densidade óptica foram realizadas a 540nm, em intervalos de 2h, por cinco dias. Os resultados foram analisados estatisticamente com o Teste de Friedman (alfa = 5%). Os melhores crescimentos foram obtidos com as linhagens M1, M2 e M3, que atingiram o início da fase log em 60 horas de cultivo. As melhores fontes de carbono variaram de acordo com a linhagem estudada, e extrato de levedura provou ser a melhor fonte de nitrogênio para todas as linhagens. Acetato de sódio inibiu o crescimento das quatro linhagens, sendo a M3 a mais afetada. O uso do sistema automatizado foi muito conveniente para as culturas em meio liquido, sendo rápido e automático, constituindo em uma boa técnica para a determinação das condições ambientais ótimas para crescimento de fungos filamentosos.

Flavobacterium plurextorum sp. nov. Isolated from Farmed Rainbow Trout (Oncorhynchus mykiss)

Five strains (1126-1H-08(T), 51B-09, 986-08, 1084B-08 and 424-08) were isolated from diseased rainbow trout. Cells were Gram-negative rods, 0.7 µm wide and 3 µm long, non-endospore-forming, catalase and oxidase positive. Colonies were circular, yellow-pigmented, smooth and entire on TGE agar after 72 hours incubation at 25°C. They grew in a temperature range between 15°C to 30°C, but they did not grow at 37°Cor 42°C. Based on 16S rRNA gene sequence analysis, the isolates belonged to the genus Flavobacterium. Strain 1126-1H-08(T) exhibited the highest levels of similarity with Flavobacterium oncorhynchi CECT 7678(T) and Flavobacterium pectinovorum DSM 6368(T) (98.5% and 97.9% sequence similarity, respectively). DNA-DNA hybridization values were 87 to 99% among the five isolates and ranged from 21 to 48% between strain 1126-1H-08(T), selected as a representative isolate, and the type strains of Flavobacterium oncorhynchi CECT 7678(T) and other phylogenetic related Flavobacterium species. The DNA G+C content of strain 1126-1H-08(T) was 33.2 mol%. The predominant respiratory quinone was MK-6 and the major fatty acids were iso-C15∶0 and C15∶0. These data were similar to those reported for Flavobacterium species. Several physiological and biochemical tests differentiated the novel bacterial strains from related Flavobacterium species. Phylogenetic, genetic and phenotypic data indicate that these strains represent a new species of the genus Flavobacterium, for which the name Flavobacterium plurextorum sp. nov. was proposed. The type strain is 1126-1H-08(T) ( = CECT 7844(T) = CCUG 60112(T)).

Influence of the vintage, clone and rootstock on the chemical characteristics of Syrah tropical wines from Brazil.

In the Northeast of Brazil, vines can produce twice a year, because annual average temperature is 26ºC, with high solar radiation and water availability for irrigation. Many cultivars have been tested according to their adaptation to the climate and soil, and the main variety used for red wines is Syrah. This work aimed to evaluate five clones of Syrah, grafted on two rootstocks, in two harvests of the second semester of 2009 and 2010, according to the chemical analyses of the wines.The clones evaluated were 100, 174, 300, 470 and 525, the rootstocks were Paulsen 1103 and IAC 313 (Golia x Vitis caribeae). Grapes were harvested in November 2009 and 2010 and the yield was evaluated. Climate characteristics of each harvest was determined and correlated to the results. Wines were elaborated in glass tanks of 20 L, with alcoholic fermentation at 25ºC for seven days, then wines were pressed and malolactic fermentation was carried out at 18ºC for 20 days. The following parameters were analyzed: alcohol content, dry extract, total anthocyanins, total phenolic index. High performance liquid chromatography was used to determine tartaric, malic, lactic and citric organic acids. Results showed that wines presented different concentrations of classical analyses, phenolics and organic acids according to the harvest date, rootstocks and clones. Principal component analysis was applied on data and clusters with wine samples were formed, explaining the variability, and results are discussed.

Influência da temperatura na germinação de sementes de Digitaria insularis, Leptochloa virgata, Pennisetum setosum e Sorghum halepense

Objetivou-se com este trabalho avaliar a influência de temperaturas constantes na germinação de sementes de espécies de plantas daninhas da família Poaceae que apresentam ocorrência em áreas de pastagem, para isto, foram analisadas as sementes de Digitaria insularis, Leptochloa virgata, Pennisetum setosum e Sorghum halepense coletadas manualmente de diversas plantas em áreas de pastagem. As sementes foram dispostas em caixas gerbox sobre folha dupla de papel "germitest", umedecido com água destilada equivalente a 2,5 vezes a massa do papel seco. As caixas foram incubadas em três germinadores verticais tipo MANGELSDORF com as temperaturas de 25ºC, 30ºC e 35ºC e fotoperíodo de 12 horas. As avaliações foram realizadas, efetuando-se a contagem diária das ocorrências germinativas, a partir da protrusão da radícula, durante 28 dias após o início do estudo. Os parâmetros avaliados foram porcentagem e índice de velocidade de germinação (IVG), efetuando-se a análise estatística pela análise de variância com comparação de médias pelo teste de Tukey a 5% de probabilidade e transformação dos resultados de porcentagem de germinação. As temperaturas que proporcionam os melhores resultados de porcentagem e velocidade de germinação são 30 e 35°C, para as espécies de D. insularis, L. virgata e S. halepense, por outro lado, a espécie de P. setosum não apresenta germinação em nenhuma das temperaturas avaliadas (25º 30º e 35ºC) não são adequadas para a sua germinação.

Imagens orbitais e termografia infravermelho na avaliação da temperatura de superfície em diferentes usos e cobertura do solo na Floresta Nacional do Tapajós e seu entorno, PA.

A Temperatura da Superfície Terrestre (TST), além de ser uma componente importante no balanço de energia na superfície, modula a temperatura do ar nas camadas mais baixas da atmosfera. O objetivo do presente trabalho foi analisar a variação da temperatura de superfície em Diferentes Usos e Cobertura do Solo na Floresta Nacional do Tapajós e seu Entorno. Utilizou-se sensores ?Thermal Infrared Sensor? dos satélites Ladsat 5 e 8 através dos ?softwares? PCI Geomatica 2015 e o QGis 2.8. Fez-se campanha de campo para obtenção de imagens termogéficas no infravermelho (câmera ThermoVision, modelo A320). Os resultados apontaram maior variabilidade espacial de TST em função da heterogeneidade do uso da terra como a agricultura anual, pecuária extensiva, dentre outros. Os resultados evidenciaram uma amplitude térmica de 13ºC na FNT+ZA. Em área de Floretas, vegetação secundária, pasto, agricultura anual as maiores variações térmicas (TST) chegaram a valores de 25ºC, 26ºC, 35ºC e 33ºC, respectivamente. A amplitude térmica na FNT foi de 5ºC e na Zona de Amortecimento de 20º, indicando perdas na cobertura vegetal, principalmente na porção nordeste da ZA. Nas áreas de proteção legal a variação de TST foi entre 19ºC a 29ºC. Conclui-se que na FLONA Tapajós a manutenção da cobertura florestal reduz as amplitudes térmicas. Por outro lado, as extensas áreas com pastagens e cultivos anuais na porção nordeste da FLONA apresentam as maiores variações espaciais da TST na Zona de Amortecimento que podem comprometer o microclima na FNT+ZA