Toxicity of silver nanoparticles on different tissues in adult Danio rerio.

Although silver nanoparticles (AgNP) are among the most studied nanomaterials by virtue of their broad application in many areas, little is known about their overall toxicity to aquatic organisms after their contamination of the water environment. This study aimed to analyze the effect of the exposure (96 h) to different AgNP concentrations on Danio rerio (zebrafish) tissues. AgNP were synthesized and characterized by transmission electron microscopy (TEM), showing spherical AgNP of 30.00 ± 16.80 nm size. The effects of different AgNP concentrations (1, 3, and 5 µg L-1) on brain, muscle, gill, and liver tissues of zebrafish were investigated. The results show a significant decrease in brain and muscle acetylcholinesterase (AChE) activity. Liver and gill catalase (CAT) activity also decreased significantly. At the highest exposure concentration, muscle AChE was more inhibited (37.3%) than brain AChE (26.4%) and gill CAT was more inhibited (67.4%) than liver CAT (51.2%). D. rerio also showed gill morphological changes such as fusion of secondary lamellae, curvature, dilated marginal channel, and epithelial lifting. This study indicates that gill CAT together with morphological studies are potential biomarkers for AgNP.

A novel de novo heterozygous ALPL nonsense mutation associated with adult hypophosphatasia.

Using genetic, clinical, biochemical, and radiographic assessment and bioinformatic approaches, we present an unusual case of adult HPP caused by a novel de novo heterozygous nonsense mutation in the alkaline phosphatase (ALPL). INTRODUCTION: Hypophosphatasia (HPP) is caused by genetic alterations of the ALPL gene, encoding the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP). Here, the purpose was to perform clinical and molecular investigation in a 36-year-old Caucasian woman suspected to present adult HPP. METHODS: Medical and dental histories were obtained for the proposita and family members, including biochemical, radiographic, and dental assessments. ALPL mutational analysis was performed by the Sanger sequencing method, and the functional impact prediction of the identified mutations was assessed by bioinformatic methods. RESULTS: We identified a novel heterozygous nonsense mutation in the ALPL gene (NM_000478.6:c.768G>A; W[TGG]>*[TGA]) associated with spontaneous vertebral fracture, severe back pain, musculoskeletal pain, low bone density, and short-rooted permanent teeth loss. Functional prediction analysis revealed that the Trp256Ter mutation led to a complete loss of TNSALP crown domain and extensive loss of other functional domains (calcium-binding domain, active site vicinity, and zinc-binding site) and over 60% loss of homodimer interface residues, suggesting that the mutant TNSALP molecules are nonfunctional and form unstable homodimers. Genotyping of the ALPL in the proposita's parents, sister, and niece revealed that in this case, HPP occurred due to a de novo mutation. CONCLUSION: The present study describes a novel genotype-phenotype and structure-function relationship for HPP, contributing to a better molecular comprehension of HPP etiology and pathophysiology.

Folha de amendoeira como aditivo em dietas para Betta splendens / Use of almond leaf as additive in diets for Betta splendens

O objetivo do presente estudo foi avaliar o desempenho zootécnico, os parâmetrosmorfométricos, o comportamento social, a viabilidade econômica e a análise microbiológica da água de cultivo e de tecidos corpóreos de Betta splendens alimentados com folha da amendoeira Terminalia catappa (FFDA) como aditivo. Foram utilizados 28 machos de Bettas, acondicionados individualmente em aquários de 1,5L, por 50 dias. O experimento foi realizado em delineamento experimental inteiramente ao acaso, com quatro tratamentos: 0,00%; 0,25%; 0,50% e 0,75% de inclusão da FFAD e sete repetições. Ao término do experimento, foram avaliados os parâmetros: desempenho zootécnico (ganho de peso diário, consumo de ração, conversão alimentar aparente, taxa de eficiência proteica, taxa de crescimento específico e fator de condição), morfométrico (comprimento total, padrão e da cabeça, altura, índice de perfil e índice de cabeça), comportamento social, viabilidade econômica da ração, análise microbiana do conteúdo intestinal, filé e escama e análise microbiológica da água. Pela ANOVA, pelo teste de Tukey e pela regressão (P>0,05), os parâmetros: peso final, ganho de peso, comprimento padrão, comprimento total e taxa de crescimento específico foram influenciados pelos tratamentos (P<0,05), apresentando um efeito quadrático. Assim, recomenda-se o nível de 0,50% de Terminalia catappa como aditivo em dietas de Betta splendens.(AU)

The objective of this study was to evaluate the performance, morphometric parameters, social behavior, economic viability, the presence of enterobacteria in the intestinal contents and a microbiological analysis of the water culture of Betta splendens fed with almond-tree-leaf flour (ATLF) as an additive. Twenty-eight male B. splendens were individually put in 1.5 L aquariums. The experiment was done in a completely randomized experimental design with four treatments: 0.00%; 0.25%; 0.50% and 0.75%, of the ATLF, dehydrated Terminalia catappa with seven repetitions each. At the end of the experimental period, the parameters were evaluated: performance (daily weight gain, feed intake, apparent feed conversion, protein efficiency rate, specific growth rate and condition factor), morphometric (total length, standard head, height, profile index and head index), social behavior, economic viability of the feed, microbial analysis of intestinal contents, fillet and scales, and microbiological analysis of the water. Though ANAVA, Tukey test and regression analysis (P> 0.05), the parameters: final weight, weight gain, standard length, total length and specific growth rate were influenced by the treatments (P< 0.05), presenting a quadratic effect. Therefore, the 0.50% level of Terminalia catappa is recommended as additive in Betta splendens diets.(AU)

Exigência de proteína bruta para juvenis de curimatã-pacu / Crude protein requeriment to curimatã-pacu juveniles

O objetivo deste estudo foi determinar a exigência de proteína bruta (PB) de juvenis de curimatã-pacu (Prochilodus argenteus). O experimento foi conduzido por um período de 60 dias, utilizando um delineamento inteiramente ao acaso, com quatro tratamentos (dieta contendo 24,0; 28,0; 32,0 e 36,0% de PB) e cinco repetições cada, em que 100 juvenis (peso inicial de 0,33 ± 0,01g) foram distribuídos em 20 aquários experimentais com volume de 130L, sendo considerada como unidade experimental uma caixa com cinco peixes, sob recirculação de água. Avaliaram-se o desempenho produtivo, o crescimento heterogêneo, os parâmetros morfométricos, fisiológicos e a viabilidade econômica das rações em função do nível de PB. Pela análise de regressão, foi verificado um aumento linear nos parâmetros de desempenho, morfométricos e fisiológicos, com o aumento do nível proteico nas rações. Quando comparados pelo teste de Tukey, o melhor resultado (P>0,05) foi obtido com 36% de PB. Verificou-se menor heterogeneidade de crescimento dos peixes à medida que se aumentavam os níveis de PB na ração. Assim, considerando também a viabilidade econômica, recomenda-se a utilização de rações com 36% de proteína bruta para juvenis de curimatã-pacu.(AU)

The aim of this study was to determinate crude protein (CP) requirement to curimatã-pacu juveniles (Prochilodus argenteus). The experiment was conducted during 60 days in a completely randomized design, with four treatments (24.0, 28.0, 32.0 and 36.0% CP of diet) and five replications, where a hundred juvenile fishes (initial weight de 0.33 ± 0.01g), were distributed in twenty tanks (130L each). Each tank with five juveniles was considered an experimental unit, with water recirculation. Performance, heterogeneous growth, morphometric and physiological parameters, and economic viability of the rations according to the CP levels were evaluated. For the regression analysis, a linear increase in performance, heterogeneous growth and morphometric and physiological parameters with increased of the protein in rations were observed. When compared by the Tukey test (P>0.05), the best results were obtained with 36% CP. There was lower fish growth heterogeneity as the levels of CP in the diet increased. Thus, taking into consideration the economic viability it is recommended to use feed containing 36% of crude protein to curimatã-pacu juveniles.(AU)

Desempenho de tambaquis (Colossoma macropomum) submetidos a restrição alimentar e a realimentação em tanques-rede / Performance of tambaqui submitted feed deprivation and refeeding in cages

Objetivou-se avaliar o desempenho de tambaquis (Colossoma macropomum) submetidos à restrição alimentar e à realimentação em tanques-rede. Durante 60 dias, foram alocados 100 tambaquis (peso inicial de 8,0 ± 0,5g), em 20 tanques-rede. Foram realizados quatro tratamentos: T1=animais alimentados diariamente (controle), T2=alimentados durante seis dias com um dia de restrição de ração (6A/1R), T3=alimentados durante cinco dias com dois dias de restrição (5A/2R) e T4 = alimentados durante quatro dias com três dias de restrição (4A/3R). Os animais foram alimentados duas vezes ao dia, às oito e às 16 horas, até a aparente saciedade, com ração extrusada comercial (42%PB). Utilizaram-se cinco repetições para cada tratamento e cinco peixes por unidade experimental, em um delineamento inteiramente ao acaso. A coleta de dados para avaliação do desempenho produtivo foi feita com base no peso final, no ganho de peso, no consumo médio de ração, na conversão alimentar, no comprimento total, no comprimento padrão, na altura, no índice de cabeça e de perfil, na taxa de eficiência proteica, na taxa de crescimento específico e na sobrevivência. Os resultados foram submetidos à ANOVA e ao teste de Tukey (P≥5). O tratamento 5A/2R apresentou-se como a melhor ferramenta na redução de custos de produção sem prejuízo ao desempenho produtivo de juvenis de tambaqui.

The aim of this research was to evaluate the performance of tambaqui juveniles (Colossoma macropomum) submitted to feed deprivation and refeeding in cages. For 60 days of experimentation, 100 tambaquis were used (initial weight 8.0 ± 0.5g), distributed in 20 cages. The treatments were: T1=daily fed animals (control), T2=fed for six days and one day of restriction (6F/1R), T3=fed for five days and two days of restriction (5F/2R) and T4=fed for four days with three days of restriction (4F/3R). With five replicates each, and five fish for each experimental unit, arranged in a completely randomized design. In accordance with treatments, the fishes were fed twice a day at 08:00am and 4:00pm. The data collection for the performance evaluation was based on final weight, weight gain, average feed intake, alimentary conversion, total length, standard length, height, index head, index profile, protein efficiency rate, specific growth rate and survival at the end of the experimental period. Results were submitted to ANOVA and Tukey test (P≥5). Treatment 5F/2R resulted in the best performance results, which is a tool in reducing production costs without loss in performance of tambaqui juvenile.

Thioester reduction and aldehyde transamination are universal steps in actinobacterial polyketide alkaloid biosynthesis.

Actinobacteria produce a variety of polyketide alkaloids with unusual structures. Recently, it was shown that a type I modular polyketide synthase (PKS) is involved in the assembly of coelimycin P1, a polyketide alkaloid produced by Streptomyces coelicolor M145. However, the mechanisms for converting the product of the PKS to coelimycin P1 remain to be elucidated. Here we show that the C-terminal thioester reductase (TR) domain of the PKS and an ω-transaminase are responsible for release of the polyketide chain as an aldehyde and its subsequent reductive amination. Bioinformatics analyses identified numerous gene clusters in actinobacterial genomes that encode modular PKSs with a C-terminal TR domain and a homolog of the ω-transaminase. These are predicted to direct the biosynthesis of both known and novel polyketide alkaloids, suggesting that reductive chain release and transamination constitutes a conserved mechanism for the biosynthesis of such metabolites.

Mechanomyography signals in spastic muscle and the correlation with the modified Ashworth scale.

The modified Ashworth scale (MAS) is the most widely used measurement technique to assess levels of spasticity. In MAS, the evaluator graduates spasticity considering his/her subjective analysis of the muscular endurance during passive stretching. Therefore, it is a subjective scale. Mechanomyography (MMG) allows registering the vibrations generated by muscle contraction and stretching events that propagate through the tissue until the surface of the skin. With this in mind, this study aimed to investigate possible correlations between MMG signal and muscle spasticity levels determined by MAS. We evaluated 34 limbs considered spastic by MAS, including upper and lower limbs of 22 individuals of both sexes. Simultaneously, the MMG signals of the spastic muscle group (agonists) were acquired. The features investigated involved, in the time domain, the median energy (MMGME) of the MMG Z-axis (perpendicular to the muscle fibers) and, in the frequency domain, the median frequency (MMGmf). The Kruskal-Wallis test (p<;0.001) determined that there were significant differences between intergroup MAS spasticity levels for MMGme. There was a high linear correlation between the MMGme and MAS (R2=0.9557) and also a high correlation as indicated by Spearman test (ρ=0.9856; p<;0.001). In spectral analysis, the Kruskal-Wallis test (p = 0.0059) showed that MMGmf did not present significant differences between MAS spasticity levels. There was moderate linear correlation between MAS and MMGmf (R2=0.4883 and Spearman test [ρ = 0.4590; p <; 0.001]). Between the two investigated features, we conclude that the median energy is the most viable feature to evaluate spasticity due to strong correlations with the MAS.

Aminas bioativas e qualidade da carne de frangos de corte / Bioactive amines and meat quality of broiler chickens

Com o objetivo de avaliar a qualidade da carne de frangos de corte mediante pesquisa dos níveis de aminas bioativas, foram coletadas, pelos serviços de inspeção oficiais, 160 amostras de carcaças provenientes de cinco regiões distintas do estado de Minas Gerais, durante o período de um ano. As poliaminas (espermidina e espermina) e as aminas biogênicas (putrescina, cadaverina, histamina, tiramina) foram pesquisadas por cromatografia líquida de alta eficiência e detecção ultravioleta (CLAE/UV). Os resultados encontrados demonstraram a presença das poliaminas, espermidina e espermina, em todas as amostras, em concentrações médias de 3,56mg/100g e 5,72mg/100g, respectivamente. Em todas as amostras foram detectadas, em concentrações muito baixas, as aminas putrescina, cadaverina, histamina e tiramina. Foi concluído que a carne de frangos de corte produzida no estado de Minas Gerais é uma fonte de poliaminas, importantes para o crescimento e a proliferação celular, e que os baixos teores de aminas biogênicas encontrados não representam riscos à saúde do consumidor, indicando que esse tipo de carne apresenta boa qualidade, tomando por base o critério de aminas bioativas.

In order to evaluate the meat quality of broiler chickens by searching the bioactive amines level, 160 samples of carcass from the five regions of the Minas Gerais State, were collected during one year by the official inspection service. The poliamines (spermidine and spermine) and the biogenic amines (putrescine, cadaverine, histamine and tyramine) were determined by high performance liquid chromatography with ultraviolet detection (HPLC/UV). The results demonstrated the presence of polyamines spermidine and spermine in all samples, at mean concentrations of 3.56mg/100g and 5.72mg/100g, respectively. The biogenic amines putrescine, cadaverine, histamine and tyramine were also found, but in low concentrations. It was concluded that the chicken broiler meat produced in Minas Gerais state is a source of polyamines, important for growth and cell proliferation; and that the biogenic amine levels found were low, and do not represent risks to consumer health, indicating that it has good quality, based on the criterion of bioactive amine.

Desempenho de alevinos de tilápia do nilo alimentados com folha de mandioca desidratada na dieta / Performance of nile tilapia fingerlings fed dehydrated cassava leaf in the diet

O objetivo foi avaliar o crescimento e a viabilidade econômica de alevinos de tilápia do Nilo (Oreochromis niloticus), alimentados com diferentes níveis de farinha da folha de mandioca desidratada na ração (FFMD). Foram utilizados 80 alevinos distribuídos em 20 recipientes de 18L, constando de sistema de recirculação de água, durante um período de 51 dias. Cem exemplares, alimentados três vezes ao dia até a saciedade, foram distribuídos aleatoriamente em um delineamento experimental inteiramente casualizado, constituindo-se de 4 tratamentos (0, 5, 10 e 15% de adição da FFMD na ração) e 5 repetições cada. Os peixes alimentados com 10% de FFMD apresentaram melhor homogeneidade no crescimento. Entretanto, os indivíduos alimentados com inclusão de 5% de FFMD apresentaram homogeneidade de crescimento similar aos alimentados com 10%. Além disso, os peixes alimentados com 5% de FFMD obtiveram melhor desempenho zootécnico e viabilidade econômica da ração. Dessa maneira, recomenda-se a utilização de 5% da farinha de folha de mandioca desidratada em rações para tilápia do Nilo.

The aim was to evaluate the growth and the economic viability of the Nile tilapia (Oreochromis niloticus) fed with different levels of dehydrated cassava leaf flour in the diet (FFMD). 80 fingerlings were distributed into 20 18L containers, consisting of a water recirculation system over a period of 51 days. 100 copies fed three times a day to satiation were randomly assigned in a completely randomized design, constituting 4 treatments (0%, 5%, 10% and 15% addition of FFMD in diet) with 5 repetitions each. The fishes fed with 10% FFMD had better homogeneity in growth. However, individuals fed with inclusion of 5% FFMD homogeneity grew similarly to those fed with 10% FFMD. In addition, the fish from the 5% had better growth performance and economic viability of the feed. Thus, it is recommended to use 5% of dehydrated cassava leaf meal in diets for Nile tilapia.

Monitoring of metals, organic compounds and coliforms in water catchment points from the Sinos River basin.

Unplanned use and occupation of the land without respecting its capacity of assimilation and environmental purification leads to the degradation of the environment and of water used for human consumption. Agricultural areas, industrial plants and urban centres developed without planning and the control of effluent discharges are the main causes of water pollution in river basins that receive all the liquid effluents produced in those places. Over the last decades, environmental management has become part of governmental agendas in search of solutions for the preservation of water quality and the restoration of already degraded resources. This study evaluated the conditions of the main watercourse of the Sinos River basin by monitoring the main physical, chemical and microbiological parameters described in the CONAMA Resolution no. 357/2005.The set of parameters evaluated at five catchment points of water human consumption revealed a river that has different characteristics in each reach, as the upper reach was class 1, whereas the middle and lower reaches of the basin were class 4. Monitoring pointed to households as the main sources of pollutants in those reaches, although metals used in the industrial production of the region were found in the samples analyzed.

Molecular characterization of Corynebacterium pseudotuberculosis isolated from goats using ERIC-PCR.

Corynebacterium pseudotuberculosis, the infectious agent of caseous lymphadenitis (CLA), is responsible for substantial economic losses in goat and sheep production. Molecular characterization of C. pseudotuberculosis isolates by enterobacterial repetitive intergenic consensus (ERIC)-PCR has shown promising results in genotyping strains isolated from sheep with CLA. We evaluated the genetic diversity of C. pseudotuberculosis isolates collected from the Sertão region of the Pernambuco (PE) State, Brazil, and investigated the potential of ERIC-PCR as a tool for the molecular typing of strains of C. pseudotuberculosis isolated from goats. Thirty-two C. pseudotuberculosis strains isolated from goats in the municipalities of Floresta and Ibimirim, PE, C. pseudotuberculosis type strain ATCC 19410, the 1002 vaccine strain, and a field isolate of Rhodococcus equi were fingerprinted using the primers ERIC-1R and ERIC-2 and the primer pair ERIC- 1R+ERIC-2. Using 100% similarity as the cutoff, 8, 10, and 7 genotypes were obtained with ERIC-1-PCR, ERIC-2-PCR, and ERIC-1+2-PCR, respectively. The Hunter-Gaston discriminatory index calculated for the ERIC-1-PCR was 0.75. The index for the ERIC-2-PCR was 0.88, and the index for the ERIC-1+2-PCR was 0.79. Among goat isolates of C. pseudotuberculosis, three, two and four genotypes (found by ERIC-1-PCR, ERIC-2-PCR, and ERIC-1+2-PCR, respectively) had been previously described among sheep isolates from Minas Gerais State, Brazil. These results showed that ERIC-PCR has good discriminatory power and typeability, making it a useful tool for discrimination among C. pseudotuberculosis isolates from goats.

Seroprevalence of Toxoplasma infection among pregnant women in Bahia, Brazil.

Infection by Toxoplasma gondii represents a risk to fetal development during pregnancy. In this study, the serological profile of pregnant women from different regions of Bahia, Brazil, was determined. Tests were conducted at LACEN-BA (Salvador, Brazil). The mean age of the women was 24.5 years (±7.41 years) and 56.4% were positive for IgG and negative for IgM specific for T. gondii. Seronegative women represented 35.9% of the total (IgG- and IgM-negative) and inconclusive results comprised 4%. Differences were observed in distinct regions. Therefore, a preventive action must be reinforced in specific regions aimed at early diagnosis to minimise the risk of congenital toxoplasmosis development.

Low sampling rates bias outcomes from the Wingate test.

The purpose of this work was to apply a simple method for acquisition of power output (PO) during the Wingate Anaerobic Test (WAnT) at a high sampling rate ( S(R)) and to compare the effect of lower S(R) on the measurements extracted from the PO. 26 male subjects underwent 2 WAnTs on a cycle ergometer. The reference PO was calculated at 30 Hz as a function of the linear velocity, the moment of inertia and the frictional load. The PO was sampled at 0.2, 0.5, 1, 2 and 5 Hz. Both the peak (16.03±2.22 W·kg (-1)) and mean PO (10.34±1.01 W·kg (-1)) presented lower relative values when the S(R) was lower. Peak PO was attenuated by 0.29-42.07% for decreasing sampling rates, resulting in different values for 0.2 and 1 Hz ( P<0.001). When the S(R) was 0.2 Hz, the time to peak was delayed by 53.81% ( P<0.001) and the fatigue index was attenuated by 22.12% ( P<0.001). In conclusion, due to the differences achieved here and the fact that the peak flywheel frequency is around 2.3 Hz, we strongly recommend that the PO be sampled at 5 Hz instead of 0.2 Hz in order to avoid biased errors and misunderstandings of the WAnT results.

Effect of kinin B2 receptor ablation on skeletal muscle development and myostatin gene expression.

Bradykinin (BK) is an active peptide that binds to the kinin B(2) receptor and induces biological events during the development and adult life. In this study we aimed to investigate the effect of kinin B(2) receptor ablation in the postnatal skeletal muscle development and body composition in adult life. For studies of skeletal muscle development, control (C57Bl6 - WT) and B(2) receptor knockout mice (B(2)(-/-)) were sacrificed at 15, 30 and 90days after birth, the gastrocnemius skeletal muscle was weighed and myostatin gene expression evaluated by real time PCR. For energy balance determination, data from control and B(2)(-/-) at 90 and 120days were collected by calorimetric method. Body composition at 120days was determined by chloroform-methanol (total body fat) and Lowry-modified method (total body protein). The results show that B(2)(-/-) have significantly increased total body weight at 15, 30 and 90days of life, when compared to WT. The weight of the gastrocnemius skeletal muscle was also significantly increased at 30 and 90days of life. Body composition analyses revealed that B(2)(-/-) mice exhibit more total corporal protein and less total corporal fat. Energy balance revealed that B(2)(-/-) have increased metabolizable energy intake and energy expenditure when compared to control mice, resulting in a lower energy gain. Interestingly, myostatin mRNA expression was significantly decreased in 15 and 30days old B(2)(-/-) mice and after icatibant treatment of WT adult mice for 5days. In conclusion, together our results show that kinin B(2) receptor deletion increases lean mass, reduces fat mass and improves metabolism efficiency in mice. The mechanism involved in this phenotype could be related to the reduction of myostatin gene expression during postnatal life.

Bradykinin-related peptides from Phyllomedusa hypochondrialis.

Bradykinin related peptides (BRPs) present in the water-soluble secretion and freshly dissected skin fragments of Phyllomedusa hypochondrialis were investigated by mass spectrometry techniques. Eighteen BRPs, along with their post-translational modifications, were characterized in the secretion by de novo MS/MS sequencing and direct MALDI imaging experiments of the frog skin. These molecules revealed strong sequence similarities to the main plasma kinin of some mammals and reptiles. Such a diversity of molecules, within the same peptide family, belonging to a single amphibian species may be related to functional specializations of these peptides and a variety of corresponding receptors that might be present in a number of different predators. Also, a novel analog, [Val]1,[Thr]6-bradykinyl-Gln,Ser had its biological activity positively detected in cell culture expressing the human bradykinin B2 receptor and in guinea pig ileum preparations.