Biomechanical and biochemical investigation of erythrocytes in late stage human leptospirosis

Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira, which can cause lipid changes in the erythrocyte membrane. Optical tweezers were used to characterize rheological changes in erythrocytes from patients with leptospirosis in the late stage. Biochemical methods were also used for quantification of plasma lipid, erythrocyte membrane lipid, and evaluation of liver function. Our data showed that the mean elastic constant of erythrocytes from patients with leptospirosis was around 67% higher than the control (healthy individuals), indicating that patient's erythrocytes were less elastic. In individuals with leptospirosis, several alterations in relation to control were observed in the plasma lipids, however, in the erythrocyte membrane, only phosphatidylcholine showed a significant difference compared to control, increasing around 41%. With respect to the evaluation of liver function of individuals with leptospirosis, there was a significant increase in levels of alanine transaminase (154%) and aspartate transaminase (150%), whereas albumin was 43.8% lower than control (P<0.01). The lecithin-cholesterol acyltransferase fractional activity was 3.6 times lower in individuals with leptospirosis than in the healthy individuals (P<0.01). The decrease of the erythrocyte elasticity may be related to the changes of erythrocyte membrane phospholipids composition caused by disturbances that occur during human leptospirosis, with phosphatidylcholine being a strong candidate in the erythrocyte rheological changes.

Análise evolutiva da morfologia e da irrigação do timo de javalis (Sus scrofa Linnaeus, 1758) / Evolutionary analysis of morphology and irrigation of the thymus of boar's (Sus scrofa Linnaeus, 1758)

Objetivou-se descrever comparativamente as artérias responsáveis pela irrigação dos lobos tímicos cervicais e torácicos dos javalis, determinando-se também as características morfológicas do referido órgão nessa espécie, a ponto de correlacioná-las evolutivamente com os demais representantes da família Suidae. Os lobos tímicos cervicais direito e esquerdo receberam ramos distribuídos pelas artérias carótidas comuns direita e esquerda e cervicais superficiais direita e esquerda e, por vezes, pelas artérias torácica externa esquerda e subclávia esquerda. Essa característica vascular foi mantida ao longo da evolução, estando presente em linhagens e raças mais recentes de suídeos. A irrigação dos lobos tímicos torácicos direito e esquerdo foi realizada predominantemente por ramos diretos e indiretos ipsilaterais e contralaterais das artérias torácicas internas direita e esquerda, e também por ramos da artéria subclávia esquerda. Ramos diretos das artérias cervicais superficiais direita e esquerda e ramos diretos da artéria torácica interna direita foram encontrados para o lobo tímico médio, sendo essa uma descrição única nessa espécie, o que demonstra uma característica evolutiva primitiva nesse ancestral.(AU)

The objective of this study was to comparatively describe the arteries responsible for the irrigation of the cervical and thoracic thymus lobes of Boars, also determining the morphological characteristics of this body in this species to the point of evolutionarily correlating them with other representatives of the Suidae family. The left and right cervical thymic lobes received branches distributed by common right and left, superficial right and left carotid arteries, and neck and, sometimes, the outer left chest and left subclavian arteries. This feature was vascular maintained throughout evolution and is present in more recent strains and breeds of swine. Irrigation of the right and left thoracic thymic lobes was performed predominantly by direct and indirect ipsilateral and contralateral branches of the right and left internal thoracic arteries and also by branches of the left subclavian artery. Direct branches of the right and left superficial cervical artery and right branches of the right internal thoracic artery were found for the average thymic lobe, this being a unique description in this species, demonstrating a primitive feature in this evolutionary ancestor.(AU)

Identification of a historic horse ecotype analyzing speech content in central Brazil / Resgate de um ecótipo equino histórico mediante análise de conteúdo de discurso no Brasil central

This study represents a first attempt to rescue germplasm of a traditional domestic equine cited in early Iberian and South American literature as Curraleiro horse. As an effort to identify and possibly characterize this type of horse we accessed traditional knowledge in strategic rural areas. Most areas were configured by hinterland settlements founded by runaway slave communities from the sixteenth century. Twenty interviews were conducted and analyzed using classical Speech Content Analysis. As a result, five response categories were recognized: 1) Identification and location of Curraleiro horses; 2) Characterization of Curraleiro horses; 3) Historical aspects; 4) Curraleiro horse conservation prospects and 5) Sanitary and productive aspects of Curraleiro horse breeding. We concluded that although scarce, the Curraleiro horse, cited in literature, still exists and showed phenotypical features which allow fine discrimination from any officially recognized breed in Brazil. Our findings may open the door for the recognition of a genuine animal resource which could play a role in rural development for specific regions and traditional communities. Speech Content Analysis was effective for in loco identification and characterization of rare traditional livestock. Thus, we encourage the usage of this tool as an alternative to subsidize identification and genetic conservation programs worldwide.(AU)

Esse estudo representa uma primeira tentativa de resgate de germoplasma em um equino doméstico tradicional citado em literatura precoce Iberiana e Sul Americana como um cavalo Curraleiro. Num esforço de identificar e possivelmente caracterizar esse tipo de cavalo, acessamos conhecimento tradicional em diversas áreas rurais estratégicas. A maioria consistia em assentamentos rurais fundados por comunidades escravas foragidas no século dezesseis. Vinte entrevistas foram conduzidas e analisadas utilizando análise de conteúdo de discurso clássico. Como resultado, cinco categorias de resposta foram reconhecidas: 1) Identificação e localização de cavalos Curraleiros; 2) Caracterização de cavalos curraleiros; 3) Aspectos históricos, 4) Perspectivas de conservação, e 5) Aspectos sanitários e produtivos da criação de cavalos Curraleiros. Concluímos que, apesar de escasso, o cavalo Curraleiro, citado na literatura, ainda existe e apresenta características fenotípicas que permitem discriminação de raças oficialmente reconhecidas no Brasil. Nossos achados podem abrir alas para o reconhecimento de um recurso animal genuíno que pode ter um papel no desenvolvimento rural em regiões específicas e comunidades tradicionais. Análise de conteúdo de discurso foi eficaz para identificação e caracterização in loco de criação tradicional rara. Desta forma, encorajamos o uso dessa ferramenta como alternativa para subsídio de identificação e programas de conservação genética ao redor do mundo.(AU)

SNPs rs471462296, rs456245081 and rs438495570 in the 5'UTR region of DGAT1 gene in Nelore / SNPs rs471462296, rs456245081 e rs438495570 na região 5'UTR do gene DGAT1 em Nelore

O objetivo desta pesquisa foi avaliar os SNPs rs471462296, rs456245081 e rs438495570 do gene DGAT1 em bovinos Nelore. Foram analisados 109 bovinos. A extração do DNA genômico foi realizada do sangue dos animais, usando-se o kit Ilustra Blood Genomic Prep Mini Spin® (GE Healthcare, UK). A concentração e o grau de pureza do DNA foram determinados por meio de espectrofotômetro (Nanodrop - Thermo Fisher Scientifc, USA). A genotipagem dos SNPs ocorreu mediante o emprego do ensaio Taqman® (Applied Biosystems, USA). Na análise genômica, não foram encontradas alterações nas frequências alélicas e genotípicas (P≥0,05) para os SNPs testados. Dessa forma, a região 5'UTR analisada apresentou-se monomórfica e a variação de SNPs não foi observada, o que limita seu uso como marcadores moleculares para o gene DGAT1 em Nelore.(AU)

Cardiac function and intracellular Ca2+ handling proteins are not impaired by high-saturated-fat diet-induced obesity

Obesity is often associated with changes in cardiac function; however, the mechanisms responsible for functional abnormalities have not yet been fully clarified. Considering the lack of information regarding high-saturated-fat diet-induced obesity, heart function, and the proteins involved in myocardial calcium (Ca2+) handling, the aim of this study was to test the hypothesis that this dietary model of obesity leads to cardiac dysfunction resulting from alterations in the regulatory proteins of intracellular Ca2+ homeostasis. Male Wistar rats were distributed into two groups: control (C, n=18; standard diet) and obese (Ob, n=19; high-saturated-fat diet), which were fed for 33 weeks. Cardiac structure and function were evaluated using echocardiographic and isolated papillary muscle analyses. Myocardial protein expressions of sarcoplasmic reticulum Ca2+-ATPase, phospholamban (PLB), PLB serine-16 phosphorylation, PLB threonine-17 phosphorylation, ryanodine receptor, calsequestrin, Na+/Ca2+ exchanger, and L-type Ca2+ channel were assessed by western blot. Obese rats presented 104% increase in the adiposity index (C: 4.5±1.4 vs Ob: 9.2±1.5%) and obesity-related comorbidities compared to control rats. The left atrium diameter (C: 5.0±0.4 vs Ob: 5.5±0.5 mm) and posterior wall shortening velocity (C: 36.7±3.4 vs Ob: 41.8±3.8 mm/s) were higher in the obese group than in the control. The papillary muscle function was similar between the groups at baseline and after inotropic and lusitropic maneuvers. Obesity did not lead to changes in myocardial Ca2+ handling proteins expression. In conclusion, the hypothesis was not confirmed, since the high-saturated-fat diet-induced obese rats did not present cardiac dysfunction or impaired intracellular Ca2+ handling proteins.

Evaluación toxicológica y de metales (Cu, Pb, Ni, Zn y Cd) en el sedimento del reservorio Paiva Castro en Mairiporã-SP, Brasil / Toxicological and metal evaluation (Cu, Pb, Ni, Zn and Cd) in the sediment of Paiva Castro reservoir, in Mairiporã-SP, Brasil

El hombre ha cambiado el ambiente para sostener la demanda global de recursos naturales como el agua. La gestión de los cuerpos hídricos tiene que ser constante, con el propósito preventivo y correctivo, dependiendo del estado de antropización de cada sistema. El objetivo de este artículo fue analizar la toxicidad del sedimento y los metales Mn, Zn, Pb, Ni, Cd y Cu, en el reservorio Paiva Castro, que abastece la Region Metropolitana de São Paulo. Se realizaron 2 muestreos (Mayo 2011, estación seca y Enero de 2012, estación húmeda) y se analizaron 5 puntos próximos a la captación de agua por la Companhia de Saneamento Básico do Estado de São Paulo. Se realizaron ensayos de toxicidad aguda y crónica en sedimento, a través de ensayos biológicos con el cladócero Daphnia similis y el insecto Chironomus xanthus. El tratamiento de datos se realizó con el test de Fisher (mortalidad). El nivel de asociación entre las variables en sedimento y en los test ecotoxicológicos fueron evaluados por test no-paramétricos, a través del coeficiente de correlación de Spearman's. Los resultados del presente trabajo señalaron bajas concentracones de metales en el sedimento del área de estudio y ausencia de toxicidad en los organismos ensayados. Se puede concluir que área estudiada del reservorio Paiva Castro se encuentra poco impactada por los metales, sin efectos directos sobre la calidad de vida los organismos bentónicos: D. similis y C. xanthus.

Man had changed the natural environment in an attempt trying to supply the global demand for resources. The management of the hydric bodies has to be constant, with preventive and corrective purpose, depending on the eutrophization state of each one. The objective of this article was to analyze the sediment toxicity and the metals Mn, Zn, Pb, Ni, Cd and Cu, in the Paiva Castro reservoir, that supply the Metropolitan Region of São Paulo. It was made 2 collections (May 2011, dry season and January 2012, wet season). It was analyzed 5 points next to the water captation station by the Companhia de Saneamento Básico do Estado de São Paulo. The sediment was analyzed as for acute and chronic toxicity through bioassays with the cladocerans Daphnia similis and the insect Chironomus xanthus. Data treatment was done with Fisher Exact Test (mortality). The association level between the variables in sediment and ecotoxicological tests was available in non-parametric tests, through the Spearman's correlation coefficient. Oriented on the results presented in this work, pointing low concentrations of heavy metals in the sediments presented in the collect local, and the absence of toxicity, we can say that at this reservoir, at least in the collect area, it's low impacted, not implicating in direct interferences in the quality of life of benthonic organisms.

Effect of energetic cost to maintain the trap for Myrmeleon brasiliensis (Neuroptera, Myrmeleontidae) in its development and adult size / Efeito do custo energético com a manutenção da armadilha de Myrmeleon brasiliensis (Neuroptera, Myrmeleontidae) no seu desenvolvimento e no tamanho dos adultos

Abstract Antlion larvae Myrmeleon brasiliensis Návas, 1914 (Neuroptera, Myrmeleontidae) are sit-and-wait predators who build traps to catch their prey. The aim of this study was to observe under laboratory conditions, how the energy cost spent on maintenance of their traps affects: the larval developmental time, time spent as a pupa, mortality rate of larvae and adult size. M. brasiliensis larvae were collected in the municipality of Aquidauana, Mato Grosso do Sul, Brazil and were individually maintained in plastic containers and subjected to two treatments. In the control treatment larvae did not have their traps disturbed while in the manipulated treatment, larvae had their traps disturbed three times a week. The experiments were followed until adult emergence. When the adults emerged, their body size (head-abdomen), anterior and posterior wing span and width were measured. Furthermore, the number of larvae that died during the experiment was recorded. The results showed that the larvae whose traps were manipulated had longer larval development time, smaller pupal development time and were smaller adults. It can be concluded that the energy expenditure spent on maintenance of the trap constructed by M. brasiliensis larvae can affect the development of negative ways, represented by a longer larval development and reduced adult size.

Resumo Efeito do custo energético com a manutenção da armadilha de Myrmeleon brasiliensis no seu desenvolvimento e no tamanho dos adultos. Larvas de formiga-leão Myrmeleon brasiliensis são predadores senta-espera que constroem armadilhas para a captura de suas presas. O objetivo deste trabalho foi observar em laboratório, como o gasto energético despendido com a manutenção dessas armadilhas afeta: o tempo de desenvolvimento larval, o tempo de pupa, a taxa de mortalidade das larvas e o tamanho dos adultos. M. brasilienses foram coletadas no município de Aquidauana, Mato Grosso do Sul, então foram individualizadas em potes plásticos e submetidas a dois tratamentos. No tratamento controle, as larvas não tiveram suas armadilhas perturbadas e no tratamento manipulado, as larvas tiveram as suas armadilhas perturbadas três vezes por semana. Os experimentos foram acompanhados até a emergência dos adultos. Quando esses emergiam, era medido o seu tamanho corporal (cabeça-abdômen), envergadura da asa anterior e posterior e largura da asa anterior e posterior. Além disso, foi contabilizado o número de larvas mortas no decorrer dos experimentos. Como resultado foi observado que as larvas manipuladas apresentaram o tempo de desenvolvimento larval maior, o tempo de pupa menor e o tamanho dos adultos foi menor. Pode-se concluir que o gasto energético despendido com a manutenção da armadilha construída pelas larvas M. brasiliensis pode afetar o seu desenvolvimento de maneiras negativas, representados pelo maior tempo de desenvolvimento larval e menor tamanho do adulto.

Classification of different degrees of adiposity in sedentary rats

In experimental studies, several parameters, such as body weight, body mass index, adiposity index, and dual-energy X-ray absorptiometry, have commonly been used to demonstrate increased adiposity and investigate the mechanisms underlying obesity and sedentary lifestyles. However, these investigations have not classified the degree of adiposity nor defined adiposity categories for rats, such as normal, overweight, and obese. The aim of the study was to characterize the degree of adiposity in rats fed a high-fat diet using cluster analysis and to create adiposity intervals in an experimental model of obesity. Thirty-day-old male Wistar rats were fed a normal (n=41) or a high-fat (n=43) diet for 15 weeks. Obesity was defined based on the adiposity index; and the degree of adiposity was evaluated using cluster analysis. Cluster analysis allowed the rats to be classified into two groups (overweight and obese). The obese group displayed significantly higher total body fat and a higher adiposity index compared with those of the overweight group. No differences in systolic blood pressure or nonesterified fatty acid, glucose, total cholesterol, or triglyceride levels were observed between the obese and overweight groups. The adiposity index of the obese group was positively correlated with final body weight, total body fat, and leptin levels. Despite the classification of sedentary rats into overweight and obese groups, it was not possible to identify differences in the comorbidities between the two groups.

Ocorrência de cryptosporidium spp. em animais exóticos de companhia no Brasil / Occurrence of cryptosporidium spp. in exotic animals raised as pets in Brazil

A infecção por algumas espécies ou genótipos de Cryptosporidium representa um risco em potencial para a saúde pública, principalmente por causa de morbidade e mortalidade em crianças de zero a cinco anos de idade e em pacientes imunodeprimidos. Embora existam alguns relatos de infecção por Cryptosporidium em animais de companhia, sua participação na epidemiologia da criptosporidiose humana é incerta, e a literatura sobre esse tema ainda é bastante escassa. O objetivo deste estudo foi determinar a ocorrência e realizar a classificação molecular de Cryptosporidium spp. em amostras fecais de animais exóticos criados como animais de estimação no Brasil. Um total de 386 amostras de seis espécies de animais foi colhido e armazenado em solução de dicromato de potássio 5% a 4°C. Os oocistos foram purificados por centrífugo-sedimentação em água/éter, seguindo-se a extração de DNA genômico e a realização da nestedPCR para amplificação de fragmento parcial do gene da subunidade 18S do rRNA. Positividade para Cryptosporidium spp. foi observada em 11,40% (44/386) das amostras. O sequenciamento de fragmentos amplificados permitiu a identificação de Cryptosporidium tyzzeri em camundongos,Cryptosporidium murisem camundongos, hamster e chinchila, Cryptosporidium parvumem chinchila, Cryptosporidiumgenótipo hamsterem hamstere Cryptosporidium sp. em porquinho-da-índia. Os resultados deste estudo mostram que há uma variedade de espécies de Cryptosporidium presentes em animais exóticos de companhia no Brasil. Os dados sugerem que esses animais podem participar da epidemiologia da criptosporidiose humana, particularmente por seu estreito convívio.

Infection by some species or genotypes of Cryptosporidium represents a potential risk to public health, mainly because of the morbidity and mortality in children from zero to five years of age and in immunocompromised patients. Although there are some reports of Cryptosporidium infection in animals raised as pets, their participation in the epidemiology of human cryptosporidiosis is uncertain and studies on this topic are still scarce. The aim of this study was to determine the occurrence, as well as to perform the molecular classification of Cryptosporidium spp. in faecal samples of exotic animals raised as pets in Brazil. A total of 386 faecal samples from six species of animals was collected and stored in a solution 5% potassium dichromate at 4°C. The oocysts were purified by centrifugal sedimentation in water-ether, followed by genomic DNA extraction and the performance of the nested-PCR to amplify a partial fragment of 18S rRNA gene. Positivity for Cryptosporidium spp. was obtained in 11.40% (44/386) of samples. The sequencing of the amplified fragments allowed the identification of Cryptosporidium tyzzeri in mice, Cryptosporidium muris in mice, hamster and chinchilla, Cryptosporidium parvum in chinchilla, Cryptosporidium hamster genotype in hamster and Cryptosporidium sp. in guinea pig. The results of this study show that there is a variety of species of Cryptosporidium present in exotic animals raised as pets in Brazil. The data suggest that these animals may have zoonotic potential and participate in the epidemiology of human cryptosporidiosis.

Microbial dynamics during azo dye degradation in a UASB reactor supplied with yeast extract

The present work aimed to investigate the microbial dynamics during the anaerobic treatment of the azo dye blue HRFL in bench scale upflow anaerobic sludge bed (UASB) reactor operated at ambient temperature. Sludge samples were collected under distinct operational phases, when the reactor were stable (low variation of color removal), to assess the effect of glucose and yeast extract as source of carbon and redox mediators, respectively. Reactors performance was evaluated based on COD (chemical oxygen demand) and color removal. The microbial dynamics were investigated by PCR-DGGE (Polimerase Chain Reaction - Denaturing Gradient of Gel Electrophoresis) technique by comparing the 16S rDNA profiles among samples. The results suggest that the composition of microorganisms changed from the beginning to the end of the reactor operation, probably in response to the presence of azo dye and/or its degradation byproducts. Despite the highest efficiency of color removal was observed in the presence of 500 mg/L of yeast extract (up to 93%), there were no differences regarding the microbial profiles that could indicate a microbial selection by the yeast extract addition. On the other hand Methosarcina barkeri was detected only in the end of operation when the best efficiencies on color removal occurred. Nevertheless the biomass selection observed in the last stages of UASB operation is probably a result of the washout of the sludge in response of accumulation of aromatic amines which led to tolerant and very active biomass that contributed to high efficiencies on color removal.

Detecção e classificação molecular de Chlamydophila psittaci em amostras fecais de aves assintomáticas / Detection and molecular characterization of Chlamydophila psittaci in asymptomatic birds

Chlamydophila psittaci é uma bactéria que causa doença respiratória ou sistêmica em aves e em seres humanos. Em vista do risco de transmissão para humanos, o objetivo deste estudo foi detectar a presença de Chlamydophila spp. em amostras de fezes ou suabes cloacais de aves assintomáticas. Foram colhidas 403 amostras fecais ou suabes cloacais, provenientes de aves domésticas, selvagens ou exóticas. As amostras foram submetidas à PCR em tempo real para C. psittaci, para amplificação de fragmento parcial do gene da subunidade 16S do rRNA, utilizando o SsoFastTM EvaGreen® Supermix (Bio-Rad) e análise da curva de dissociação. Para determinação do genótipo de C. psittaci, foi usada a hemi-nested PCR visando à amplificação de fragmento parcial do gene OMP-A, realizada nas amostras positivas pela PCR em tempo real, seguida de sequenciamento dos fragmentos amplificados. A PCR em tempo real revelou positividade em 17 (4,21%) amostras. A hemi-nested foi positiva em 2 amostras positivas pela PCR em tempo real. O genótipo A de C. psittaci foi identificado pelo sequenciamento de uma amostra amplificada pela hemi-nested PCR. Os resultados deste experimento demonstram que a PCR em tempo real, visando à amplificação de fragmento parcial da subunidade 16S do rRNA, seguida da análise da curva de dissociação, pode ser utilizada para detecção de DNA de Chlamydophila sp. em amostras fecais de aves assintomáticas. A classificação da espécie de Chlamydophila e do genótipo de C. psittaci deve ser realizada por meio de PCR tendo como alvo o gene ompA e sequenciamento dos fragmentos amplificados.

Chlamydophila psittaci is a bacterium that causes respiratory or systemic disease in birds and humans. Owing to the risk of transmission from asymptomatic birds to humans, the objective of this study was to detect the presence of Chlamydophila spp. in asymptomatic birds. Four hundred and three fecal samples or cloacal swabs were collected from domestic, wild or exotic birds. The 403 samples were examined by real time PCR specific for the 16S subunit of rRNA gene using SsoFastEvaGreen®SupermixTM (Bio-Rad) and melting curve analysis. Hemi-nested PCR specific for the OMP-A gene, accomplished in real-time PCR positive samples, was followed by sequencing of the amplified fragments to determine the genotype of C. psittaci. Real-time PCR was positive in 17 (4.21%) samples. Hemi-nested PCR revealed positivity in two samples previously positive by real-time PCR. Sequencing of the fragment amplified by hemi-nested PCR allowed for the identification of genotype A of C. psittaci in one sample. The results of this experiment show that the real-time PCR targeting the 16S rRNA gene followed by melting curve analysis can be used for diagnosis of Chlamydophila sp. in fecal samples of asymptomatic birds. The classification of the Chlamydophila species and the genotype of C. psittaci must be accomplished by PCR targeting the ompA gene and sequencing of the amplified fragments.

Efeitos do sombreamento na anatomia foliar de Gallesia integrifolia (Spreng) Harms e Schinnus terebinthifolius Raddi / Effects of shading on leaf anatomy of Gallesia integrifolia (Spreng) Harms and Schinnus terebinthifolius Raddi

Realizou-se estudo com o objetivo de analisar as alterações na anatomia foliar de Gallesia integrifolia (Spreng) Harms e Schinnus terebinthifolius Raddi quando cultivadas em ambientes de sombra moderada ou densa, simulando as condições naturais encontradas em sistemas agroflorestais tradicionais do sul da Bahia, Brasil. Plantas das duas espécies, com aproximadamente um ano de idade, foram cultivadas em casa de vegetação sob quatro níveis de sombreamento (25%, 17%, 10% e 5%). Estudos anatômicos do limbo foliar foram realizados a partir de material incluído em parafina e seccionado em micrótomo rotativo. Os diferentes níveis de sombreamento ocasionaram alterações na estrutura do mesofilo de ambas as espécies, com diferenças significativas na espessura do parênquima paliçádico, limbo foliar, e densidade estomática. Nas condições em que o experimento foi realizado os resultados obtidos indicaram que G. integrifolia apresenta maior capacidade de aclimatação a ambientes de sombra moderada e densa do que S. terebinthifolius, sendo mais indicada para o cultivo em sistemas agroflorestais pré-estabelecidos.

This study was conducted with the aim of analyzing the changes in the leaf anatomy of Gallesia integrifolia (Spreng) Harms and Schinnus terebinthifolius Raddi when grown in environments with moderate to dense shade, simulating the natural conditions found in traditional agroforestry systems in southern Bahia, Brazil. Plants of both species, being approximately one year-old, were grown at four irradiance levels (25%, 17%, 10% and 5%) in a greenhouse. Leaf anatomical studies were made from material embedded in paraffin and sectioned on a rotary microtome. The different irradiance levels caused a shift in the mesophyll anatomy of both species, with significant differences in palisade parenchyma thickness and leaf stomatal density. In the conditions of this experiment, the results indicated that G. integrifolia has a higher capacity for acclimatization to moderate to dense shade than S. terebinthifolius, and it is thus more suitable for cultivation in pre-established agroforestry systems.

Detecção dos genes de Staphylococcus aureus, enterotoxinas e de resistência à meticilina em leite / Detection of genes of Staphylococcus aureus, enterotoxins and methicillin resistance in milk

Realizou-se a detecção do gene de Staphylococcus aureus, de enterotoxinas e de resistência à meticilina com extração de DNA feita diretamente de amostras de leite. Das 200 amostras estudadas, 145 (72,5%) amplificaram o gene femA, e estas foram analisadas quanto à presença dos genes sea, seb, sec e mecA. Os genes das enterotoxinas mais prevalentes foram: sea (60%), seb (37,9%) e sec (6,9%). Foram encontradas 18 amostras de leite (11,0 %) com S. aureus portadores do gene mecA. A detecção de S. aureus diretamente do leite, sem a necessidade de isolamento bacteriano e a caracterização do potencial enterotoxigênico, demonstra que a técnica de PCR é muito útil para estudos epidemiológicos das infecções estafilocócicas da glândula mamária. O alto percentual (72,5%) de amostras de leite positivas para a presença do gene femA sugere que S. aureus constitui um dos principais agentes causadores de infecções intramamárias na microrregião de Sete Lagoas-MG e que seu potencial enterotoxigênico e presença do gene mecA, que identifica o S. aureus resistente à meticlina, representa um risco potencial à saúde pública.

This work was performed to detect the Staphylococcus aureus gene, enterotoxins resistance to methicillin with the extraction of DNA directly from milk samples. Of the 200 samples studied 145 (72.5%) amplified the femA gene, which were analyzed regarding the presence of sea, seb, sec and mecA genes. The most prevalent enterotoxins genes were: sea (60%), seb (37.9%) and sec (6.9%). 18 milk samples (11 %) had S. aureus carrying the mecA gene. The detection of S. aureus directly from the milk, with no need for bacterial isolation and the characterization of the enterotoxigenic potential demonstrate that the PCR technique is very useful for epidemiological studies of staphylococcal infections of the mammary gland. The high percentage (72.5%) of positive milk samples for the presence of the femA gene suggests that S. aureus constitutes of the main agents which cause intramammary infections in the micro region of Sete Lagoas-MG and that its enterotoxigenic potential and the presence of the mecA gene, which identifies the S. aureus resistant to methicillin, represent a potential risk to public health.

Sensibilidade da PCR na amplificação do DNA bovino em diluição seriada e mistura de amostra macho e fêmea / Sensitivity of PCR to amplify sex-specific bovine DNA in diluted samples

A mixture of bovine DNA from a male and a female Jersey (Bos taurus taurus) bred in different proportions was used to determine the sensitivity of PCR to amplify and discriminate the bovine DNA samples. Samples were obtained from the peripheral blood of a bull and a heifer and DNA was isolated using a commercial kit for extraction and purification of nucleic acids. Two primers sets were designed to flank genomic regions: one autosomal and one Y-specific. DNA samples were diluted in water to a final concentration of 4x10-14 ng. The results showed positive amplification of the samples diluted to a concentration of 4x10-10ng and 4x10-4ng for the autosomal and Y-specific regions, respectively. PCR was able to discriminate the male DNA in a mixture of 99:1 (DNA ♀: DNA ♂) heifer to bull ratio. Therefore, the PCR was successful in amplifying the bovine genome in samples containing low concentrations of DNA. Thus, PCR can be used as a sensitive and efficient tool to determine the sex of the fetus in pregnant cows, helping to promote correct and efficient animal management, sex selection, and breeding in commercial herds.

Uso de pinças ópticas para avaliação do potencial zeta de hemácias em EDTA, CPD-SAGM e estocadas / Use of optical tweezers for evaluation of szeta potential of red blood cells in EDTA, CPD-SAGM and stored in blood banks

A hemácia é carregada negativamente, principalmente devido ao ácido siálico que gera um potencial elétrico denominado Potencial Zeta que impede a aglutinação intravascular. Os testes de hemaglutinação na rotina transfusional, necessitam de substâncias potencializadoras, das quais muitas agem diminuindo o Potencial Zeta para se ter maior sensibilidade. Através da pinça óptica, ferramenta capaz de capturar células utilizando a luz, foi proposta uma metodologia para quantificar o potencial zeta e aplicar em hemácias coletadas com EDTA e estocadas em CPD-SAGM (visando avaliar alterações de cargas da membrana relacionadas a lesões de armazenamento. Os potenciais zeta em CPD-CAGM foram superiores (-14,8 mV) aos em EDTA (-7,9 mV) e decrescentes a partir do primeiro dia de armazenamento, estabilizando-se a partir da terceira semana com potencial zeta -7,6 mV. Hemácias com CPD-SAGM apresentaram potencial zeta maior, pois possivelmente este conservante evitou lesões mais significativas da membrana que poderiam alterar as cargas. A redução do potencial zeta no armazenamento pode ser consequência de enzimas liberadas de leucócitos lisados que tenham alterado as glicoforinas da membrana. A metodologia permitiu avaliar o potencial zeta em diferentes condições e poderá contribuir na padronização de técnicas de hemaglutinação com diferentes meios potencializadores e no melhor conhecimento das lesões de estocagem para fins transfusionais.

Digestibilidade e metabólitos sanguíneos de vacas da raça Holandesa superovuladas que receberam Lac100® ou linhaça em grão como fontes de gordura / Digestibility and blood concentration of metabolites in superovulated dairy cows fed Lac100® or linseed as fat sources

Foram avaliados os efeitos de duas fontes de gordura, Lac100® (Yakult), fonte de ácido graxo ômega-6, e linhaça em grão (Linum usitatissimum) (LIN), fonte de ácido graxo ômega-3, sobre a digestibilidade aparente da matéria seca (DMS), proteína bruta (DPB), fibra em detergente neutro (DFDN), fibra em detergente ácido (DFDA) e extrato etéreo (DEE), e concentrações sanguíneas de HDL, LDL, VLDL, colesterol, triglicerídeos, glicose, cálcio, fósforo, N-ureico e progesterona. Foram utilizadas 10 vacas da raça Holandesa em lactação, distribuídas nos dois tratamentos, em um delineamento estatístico inteiramente casualizado. Os animais alimentados com Lac100® apresentaram maiores valores de DFDN (57,44 por cento vs. 50,80 por cento) e DEE (77,70 por cento vs. 72,18 por cento) quando comparados aos alimentados com LIN. Os animais alimentados com Lac100® apresentaram maiores concentrações de LDL (111,00 vs. 45,46mg/dL), HDL (82,27 vs. 64,93mg/dL) e colesterol total (203,60 vs. 116,13mg/dL) e menor concentração de N-ureico (13,22 vs. 15,19mg/dL) em relação aos animais alimentados com linhaça em grão. As concentrações sanguíneas de glicose, cálcio, fósforo e progesterona não foram alteradas. Os resultados sugerem que a fonte de gordura na dieta modifica a DFDN e a DEE e, as concentrações sanguíneas de LDL, HDL e colesterol total em vacas em lactação.

The effect of two sources of fat, calcium salts of soybean oil, Lac100® (Yakult), a source of omega-6 fatty acids or whole flaxseed (Linum usitatissiumum), a source of omega-3 fatty acids were evaluated to determine the apparent digestibility of dry matter (DDM), crude protein (DCP), neutral detergent fiber (DNDF), acid detergent fiber (DADF), and ether extract (DEE); and blood concentrations of HDL, LDL, VLDL, cholesterol, triglycerides, glucose, calcium, phosphorus, N-urea, and progesterone. Ten lactating Holstein cows were allocated in two treatments and a completely randomized design was used. Cows fed Lac100® had higher values of DNDF (57.44 percent vs. 50.80 percent) and DEE (77.70 percent vs. 72.18 percent) compared to those fed flaxseed. Cows fed Lac100® had higher blood concentrations of LDL (111.00 vs. 45.46mg/dL), HDL (82.27 vs. 64.93mg/dL), and total cholesterol (203.60 vs. 116.13mg/dL) and lower N-urea (13.22 vs. 15.19mg/dL) compared to those fed flaxseed. Blood concentrations of glucose, calcium, phosphorus, and progesterone were similar between the treatments. These results suggest that dietary fat modifies DNDF and DEE and blood concentrations of LDL, HDL, VLDL, and total cholesterol in lactating dairy cows.

Inclusão de silagem de rama de mandioca em substituição à pastagem na alimentação de vacas em lactação: produção, qualidade do leite e da gordura / Substitution of pasture by cassava foliage silage in the diet of dairy cows: production and quality of milk and milkfat

Avaliou-se o efeito da substituição da pastagem pela silagem de rama de mandioca (SRM) sobre a produção e constituintes do leite e sobre a qualidade da gordura do leite de vacas da raça Holandesa. O delineamento experimental utilizado foi um duplo quadrado latino 4x4, e para o fator níveis de suplementação, foi usado regressão. A dieta-controle foi 50 por cento concentrado e 50 por cento volumoso, e as proporções de substituição do volumoso foram 0, 20, 40 e 60 por cento com base na matéria seca. Todas as dietas eram isoprotéicas. Foram analisados: produção de leite (PL), PL corrigida a 4 por cento (PLC), acidez, gordura ( por cento), densidade, proteína, lactose, sólidos totais, contagem de células somáticas (CCS), ureia e perfil dos ácidos graxos da gordura do leite. Houve efeito linear decrescente da substituição do volumoso pela SRM sobre a PL, PLC e ureia no leite, e efeito (P<0,05) linear crescente sobre a concentração de ácido gama linolênico e ácido palmítico. A acidez, densidade, proteína, gordura, lactose, sólidos totais, CCS e demais ácidos graxos da gordura do leite permaneceram inalterados com a incorporação da SRM na dieta.

The effect of a partial substitution of pasture for cassava foliage silage (CFS) on milk production and composition on fat quality of Holstein cow milk was studied using a replicated 4 X 4 Latin square design with four treatments. Regression was used for the comparison of nutrition levels. The control diet contained 50 percent pasture and 50 percent concentrate on a dry matter basis. The substitution levels of pasture for CFS were 0, 20, 40, or 60 percent of the forage dry matter. All diets were isonitrogenous. The analyzed parameters were: milk production (MP); 4 percent fat-corrected milk production (FCM); acidity; density; concentrations of fat, protein, lactose, urea, and total solids; somatic cell counts (SCC), and milk fatty acids profile. There was a linear decrease effect (P<0.05) in MP, FCM, and urea content with increasing levels of CFS in the diet. Concentrations of gamma linolenic and palmitic acids in milk fat linearly increased (P<0.05) with higher proportions of CFS in the diet. Other contents of milk fatty acids remained similar among treatments as well as acidity; density; concentrations of protein, fat, lactose, and total solids; and SCC.

The alga Bryothamnion seaforthii contains carbohydrates with antinociceptive activity

Bryothamnion seaforthii, a red alga common to the Northeastern coast of Brazil, was used to prepare the protein fraction F0/60 by ammonium sulfate precipitation. The chromatography of F0/60 on DEAE-Sephadel column resulted in two lectin fractions, PI and PII, which have antinociceptive properties in rodents. We determined the antinociceptive activity of the PII fraction and of a carbohydrate-containing fraction (CF) in mice. The CF was prepared from the dried algae, after digestion with 100 mM sodium acetate, pH 6.0, containing 5 mM cysteine, EDTA and 0.4 percent papain, at 60ºC. A 10 percent cetylpyridinium chloride was added to the filtrate, and the precipitate was dissolved with 2 M NaCl:ethanol (100:15, v/v) followed by the carbohydrate precipitation with ethanol. The final precipitate, in acetone, was dried at 25ºC. The PII fraction markedly inhibited acetic acid-induced abdominal writhing after ip administration (control: 27.1 ± 2.20; PII 0.1 mg/kg: 5.5 ± 1.85; 1 mg/kg: 1.6 ± 0.72 writhes/20 min) and after oral administration (control: 32.0 ± 3.32; PII 0.1 mg/kg: 13.1 ± 2.50; 1 mg/kg: 9.4 ± 3.96 writhes/20 min). PII was also effective against both phases of pain induced by 1 percent formalin (control, ip: 48.2 ± 2.40 and 27.7 ± 2.56 s; PII: 1 mg/kg, ip: 34.3 ± 5.13 and 5.6 ± 2.14 s; control, po: 44.5 ± 3.52 and 25.6 ± 2.39 s; PII 5 mg/kg, po: 26.5 ± 4.67 and 15.3 ± 3.54 s for the 1st and 2nd phases, respectively) and in the hot-plate test. The CF (ip) also displayed significant antinociceptive properties in all tests but at higher doses (1 and 5 mg/kg, ip and po). Thus, CF at the dose of 5 mg/kg significantly inhibited writhes (ip: 7.1 ± 2.47 and po: 14.5 ± 2.40 writhes/20 min) as well as the 1st (po: 19.6 ± 1.74 s) and 2nd (po: 7.1 ± 2.24 s) phases of the formalin test compared to controls ip and po. The antinociceptive effects of both the PII and CF in the formalin and hot-plate tests were prevented at least partially by pretreatment with the opioid receptor antagonist naloxone (2 mg/kg, sc). Moreover, both fractions retained antinociceptive activity in the acetic acid-induced writhing test following heating, a procedure which abolished the hemagglutinating activity of the fraction, presumably due to lectins also present. Finally, both fractions also prolonged the barbiturate-induced sleeping time...

Freqüencia e assimetria da displasia coxofemoral em cäes Pastor-Alemäo / Frequency and asymmetry of hip dysplasia in German Shepherd dog

Avaliaram-se a freqüência e a assimetria da displasia coxofemoral em 181 radiografias de cäes da raça Pastor-Alemäo, de ambos os sexos, no período de junho de 1986 a março de 1993. As radiografias foram feitas na posiçäo ventrodorsal, com os membros paralelos e a pélvis simétrica. A idade média dos animais era de 16,6 meses. Observaram-se 72,4 por cento de cäes displásicos, sendo 5,3 por cento unilateral, 89,4 por cento bilateral simétrica e 5,3 por cento bilateral assimétrica em graus diferentes

Familial adenomatous polyposis.

Familial adenomatous polyposis (FAP) is a rare disorder associated with less than 1% of colorectal carcinomas (CRCs). Since FAP is a potentially preventable cause of CRC clinicians should have an adequate knowledge of it to identify the disease and to manage the patient and family. FAP is an autosomal dominant inherited disorder characterised by the development of more than a hundred adenomatous polyps in the colon and rectum which can undergo malignant change. Children of an affected individual are at 50% risk of inheriting the predisposing gene. After the identification of an index patient, genetic testing in combination with the detection of extra-colonic manifestations allows more accurate identification of family members likely to have the faulty gene, enabling the targetting of screening and preventive surgery only to those at risk. FAP also provides insights into the development, progression and prevention of sporadic CRC.