Total Mercury Determination in Muscle and Liver Tissue Samples from Brazilian Amazon Fish Using Slurry Sampling

This paper presents a slurry sampling method for total mercury determination by graphite furnace atomic absorption spectrometry (GFAAS) in tissue of fish from the Amazon. The tissue samples were lyophilized and macerated, and then the slurry samples were prepared by putting 20 mg of tissue, added to a solution containing Triton X-100, Suprapur HNO3, and zirconium nitrate directly in sampling vials of a spectrometer. Mercury standard solutions were prepared under the same conditions as the slurry samples. The slurry samples and the mercury standard solutions were sonicated for 20 s. Twenty microliters of slurry samples were injected into the graphite tube, which contained an internal wall lined with tungsten carbide. Under these conditions, it was possible to thermally stabilize the mercury up to an atomization temperature of 1700 °C. The method was validated by mercury determination in reference materials DORM-4 and DOLT-4. The LOD and LOQ were 0.014 and 0.045 mg kg−1, respectively, and recovery percentages in relation to the concentration values were certified in the order of 98%

Feedlot diets containing different starch levels and additives change the cecal proteome involved in cattle’s energy metabolism and inflammatory response

Diets for feedlot cattle must be a higher energy density, entailing high fermentable carbohydrate content. Feed additives are needed to reduce possible metabolic disorders. This study aimed to analyze the post-rumen effects of different levels of starch (25%, 35%, and 45%) and additives (monensin or a blend of essential oils and exogenous α-amylase) in diets for Nellore feedlot cattle. The cecum tissue proteome was analyzed via two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and then differentially expressed protein spots were identified with liquid chromatography–tandem mass spectrometry (LC–MS/MS). The use of blends of essential oils associated with α-amylase as a feed additive promoted the upregulation of enzymes such as triosephosphate isomerase, phosphoglycerate mutase, alpha-enolase, beta-enolase, fructosebisphosphate aldolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), l-lactate dehydrogenase B, l-lactate dehydrogenase A chain, l-lactate dehydrogenase, and ATP synthase subunit beta, which promote the degradation of carbohydrates in the glycolysis and gluconeogenesis pathways and oxidative phosphorylation, support pyruvate metabolism through the synthesis of lactate from pyruvate, and participate in the electron transport chain, producing ATP from ADP in the presence of a proton gradient across the membrane. The absence of proteins related to inflammation processes (leukocyte elastase inhibitors) in the cecum tissues of animals fed essential oils and amylase may be because feed enzymes can remain active in the intestine and aid in the digestion of nutrients that escape rumen fermentation; conversely, the effect of monensin is more evident in the rumen and less than 10% results in post-ruminal action, corroborating the hypothesis that ionophore antibiotics have a limited effect on the microbiota and intestinal fermentation of ruminants. However, the increase in starch in these diets promoted a downregulation of enzymes linked to carbohydrate degradation, probably caused by damage to the cecum epithelium due to increased responses linked to inflammatory injuries

The Genome of Anopheles darlingi, the main neotropical malaria vector

Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors ∼100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vectorhuman and vectorparasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible at www.labinfo.lncc.br/index.php/anopheles- darlingi. © 2013 The Author(s)

Oxigênio dissolvido (OD), demanda bioquímica de oxigênio (DBO) e demanda química de oxigênio (DQO) como parâmetros de poluição no ribeirão Lavapés/Botucatu - SP

Oxigênio dissolvido (OD), demanda bioquímica do oxigênio (DBO) e demanda química do oxigênio (DQO) foram utilizados como parâmetros para se avaliar o grau e capacidade de autodepuração do ribeirão Lavapés, que atravessa a cidade de Botucatu - SP. Avaliou-se o perfil de poluição orgânica e identificadas as zonas de autodepuração, e pontualmente foi realizado uma coleta de 24 horas, de hora em hora, onde foi possível correlacionar a poluição orgânica com as atividades domésticas. A DQO mostrou-se como a melhor técnica para avaliar o perfil de poluição orgânica, identificar as zonas de depuração, e para avaliar a poluição orgânica, neste curso d água. A relação DQO/DBO foi em média 3,4 caracterizando um esgoto biodegradável, indicando que praticamente não existe adição de efluentes industriais, permitindo assim estimar a DBO através da DQO. Nos trechos de água limpa, nascente e foz, em que a DQO estava abaixo de 5 mg L-1 O2, foram utilizados os valores de oxigênio consumido (método do permanganato), o que não invalidou a identificação das zonas de depuração. No entanto pesquisas para desenvolver o método de DQO (dicromato) para baixas concentrações, abaixo de 5 mg L-1 são necessárias para uma melhor avaliação da recuperação do curso d água.To evaluate the depuration capacity of the Lavapés brook, which crosses Botucatu - SP, it was used dissolved oxygen (DO), biochemical oxygen demand (BOD) and chemical oxygen demand (COD) as pollution parameters. The organic pollution profile was evaluated, the depuration zone identified and hourly 24-hour collection made, the last making it possible to correlate the organic pollution with household chores. Chemical oxygen demand (COD) showed itself to be the best technique to evaluate the organic pollution profile, to identify the depuration zones and to evaluate the organic pollution punctually in this water course. Characterizing a biodegradable sewage and indicating there that is no practical addition of industrial effluents, the average relation COD/BOD was 3.4, thus making it possible to estimate the BOD by the COD. At the clean water sections, source and mouth, where de COD was below 5 mg L-1 O2 it was used the consumed oxygen values (permanganate method), which did not invalidate the depuration zones identification. However, it is necessary researches to develop the method dichromate for low levels (below 5 mg L-1 ) in order to evaluate the water stream recuperation better

Contribuição da cidade de Botucatu - SP com nutrientes (fósforo e nitrogênio) na eutrofização da represa de Barra Bonita

As concentrações de nutrientes (fósforo e nitrogênio) na nascente, cidade e foz do ribeirão Lavapés/ Botucatu-SP foram avaliadas e comparadas com as do rio Capivara. O rio Capivara como o ribeirão Lavapés, possui nascente no alto da cuesta e desemboca suas águas na represa de Barra Bonita/ rio Tietê (Figura 1). O rio Capivara possui uma das nascentes próxima a uma do ribeirão Lavapés e, ambos possuem a foz próxima uma da outra na represa de Barra Bonita. O uso e tipo de solo nas margens de ambos cursos d´águas são bastante parecidos, o que implica em uma poluição rural semelhante, assim a grande diferença que afeta a qualidade da água, entre os dois cursos d´águas é a poluição urbana, na cidade de Botucatu (ribeirão Lavapés). Deste modo procurou-se avaliar a contribuição de nutrientes (fósforo e nitrogênio) do ribeirão Lavapés e rio Capivara na represa de Barra Bonita , e estimar a carga retida no lodo do ribeirão Lavapés e a despejada na represa em função dos lançamentos de esgotos sanitários no mesmo, na cidade de Botucatu. O trabalho foi realizado em um período de seca, sem alagamentos, o que permitiu medir as vazões próximas da foz e avaliar a carga diária, no período, de nutrientes lançados na represa. Foram avaliados também, oxigênio dissolvido, demanda química do oxigênio e condutividade Elétrica. Embora os resultados sejam estimativos e variáveis no tempo, podemos concluir que a poluição na cidade de Botucatu por esgoto sanitário além de inviabilizar o uso da água no seu percurso, e causar outros prejuízos, contribui para agravar a eutrofização na represa de Barra Bonita.It was compared the level content of nutrients (phosphorus and nitrogen) at the Lavapés brook with level content at source an cross-tow (domestic sewage discharge) and also with the level/ content at source and mouth the Capivari river. The head-waters of the Capivari river, as well as those of the Lavapés brook, are on the highlands (at the top the sierra/ mountain-range) and its waters flow into the Barra Bonita dan on the Tietê river (Figure 1). One of its sources is near a Lavapés source and their mouths are close to each other at the Barra Bonita dam. Along the banks of both streams the use of land and the types of soil are alike, which implies in a similar rural pollution; so, what makes the difference in water quality between them is the urban pollution affecting the Lavapés brook in Botucatu. Therefore it was evaluated the contribution of nutrients (phosphorus and nitrogen) to the Barra Bonita Dam of the Capivari river and the Lavapés brook. Taking into account the domestic sewage carried into the Lavapés brrok in Botucatu, it was estimated the nutrient load of its silt (kept in its silt) and the one discharged into the Barra Bonita dam. The study- was conducted during drought, without overflows, which made it possible to measure the near-mouth-flow and to evaluate the nutrient load daily discharged into the dam. It was also evaluated the dissolved oxygen, the chemical oxygen demand and the electrical conductivity. Although the results are estimated and variabel in time we can conclude that pollution of the Lavapés brook waters with domestic sewage in Botucatu, besides making the use of its waters not viable and causing other (environmental) damage (financial losses), contributes to aggravating the eutrophication at the Barra Bonita dam

Ultrasound-assisted extraction of Na and K from swine feed and its application in a digestibility assay: A green analytical procedure

The study is aimed to evaluate the efficiency of ultrasound-assisted extraction (UAE) as a simple strategy focused on sample preparation for metal determination in biological samples. The extraction of sodium and potassium extraction was carried out from swine feed followed by determination of the concentration of these metals by flame atomic emission spectrometry (FAES). The experiment was performed to cover the study of the variables influencing the extraction process and its optimal conditions (sample mass, particle size, acid concentration, sonication time and ultrasound power); the determination of these analytical characteristics and method validation using certified reference material; and the analysis of pre-starter diets. The optimal conditions established conditions were as follows: mass: 100 mg, particle size:<60 μm, acid concentration: 0.10 mol L-1 HCl, sonication time: 50 s and ultrasound power: 102 W. The proposed method (UAE) was applied in digestibility assays of those nutrients present in different piglet pre-starter feeds and their results proved to be compatible with those obtained from mineralized samples (P < 0.05). The ultrasound extraction method was demonstrated to be an excellent alternative for handless sampling and operational costs and the method also has the advantage of does not generating toxic residues that may negatively affect human health and contaminate the environment. © 2013 Elsevier B.V. All rights reserved

Determination of chromium by GFAAS in slurries of fish feces to estimate the apparent digestibility of nutrients in feed used in pisciculture

This paper presents a simple, fast and sensitive method to determine chromic oxide (used as a biological marker of fish feed) in samples of fish feces by GFAAS through the direct introduction of slurries of the samples into the spectrometer's graphite tube. The standard samples of feces and of fish feed containing 0.10-1.00 mg kg(-1) of Cr2O3 were pre-frozen for I min in liquid nitrogen and then ground a cryogenic mill for 2 min, which reduced the samples' grain size to less than 60 mu m. The standard slurries were prepared by mixing 20 mg of standard samples of fish feed or feces with I mL of a solution containing 0.05% (v/v) of Triton X-100 and 0.50% (v/v) of suprapure HNO3 directly in the spectrometer's automatic sampling glass. The final concentrations of Cr2O3 present in the standard slurries were 2, 4, 8, 16 and 20 mu g L-1. After sonicating the mixture for 20s, 10 mu L of standard slurries were injected into the graphite tube, whose internal wall was lined with a metallic palladium film that acted as a permanent chemical modifier. The limits of detection (LOD) and quantification (LOQ) calculated for 20 readings of the blank of the standard slurries (2%, m/v of feces or feed devoid of minerals) were 0.81 and 2.70 mu g L-1 of Cr2O3 for the standard feces slurries, 0.84 and 2.83 mu g L-1 of Cr2O3 for the standard feed slurries. The proposed method was applied in studies of nutrient digestibility of different fish feeds and its results proved compatible with the results obtained from samples pre-mineralized by acid digestion. (c) 2005 Elsevier B.V. All rights reserved

Disponibilidade aparente do fósforo em alimentos vegetais e suplementação da enzima fitase para tilápia-do-nilo

Este estudo foi realizado com o objetivo de avaliar o efeito da fitase (0, 1.000 e 2.000 uf/kg) na disponibilidade aparente de P de cinco alimentos energéticos (milho, milho extrusado, farelo de trigo, farelo de arroz e sorgo baixo tanino) e cinco alimentos protéicos (soja extrusada, farelo de soja, farelo de girassol, farelo de algodão e glúten de milho) utilizando-se uma ração purificada como controle. Foram utilizados 100 juvenis (100,00 ± 5,00 g) de tilápia-do-nilo (Oreochromis niloticus) alojados em dez gaiolas para manejo de alimentação e coleta de fezes. Para cada alimento e cada nível de fitase avaliado, foram coletadas amostras em cinco gaiolas, totalizando cinco repetições por tratamento. Os resultados obtidos indicaram que a suplementação de até 2.000 uf/kg de fitase não aumentou a disponibilidade aparente de P do farelo de trigo e do farelo de algodão. Entretanto, o nível de 1.000 uf/kg aumentou a disponibilidade aparente de P da soja extrusada e do farelo de girassol, enquanto o nível de 2.000 uf/kg proporcionou essa resposta para o farelo de milho, o milho extrusado, o sorgo baixo tanino, o farelo de arroz, o farelo de soja e o glúten de milho.This study was carried out to evaluate effects of phytase (0, 1,000, and 2,000 uf/kg) on apparent P availability of five energetic feeds (corn, extruded corn, wheat meal, rice meal and low-tannin sorghum) and five protein feeds (extruded soy, soybean meal, sunflower meal, cottonseed meal and corn gluten) using a purified diet as control. One hundred (100.00 ± 5.00 g) Nile tilapia juveniles (Oreochromis niloticus) were allotted to ten cages to allow feeding and fecal collection. For each feed and each phytase level evaluated samples were collected from five cages, in a total of five replications per treatment. The obtained results showed that the phytase supplementation up to 2,000 uf/kg did not increase apparent P availability present in wheat meal or cottonseed meal. However, the level of 1,000 uf/kg increased the apparent P availability present in extruded soy and sunflower meal, while the level of 2,000 uf/kg promoted this response for corn meal, extruded corn, low-tannin sorghum, rice meal, soybean meal and corn gluten