Inferências carioevolutivas sobre grupos crípticos de peixes marinhos e estuarinos

Cytogenetic studies have been revealing a great diversity not detected, until then, in several families of fishes. Many of these groups, especially those that exhibit great diversity, like Perciformes and Siluriformes, possess species with difficult morphologic characterization, called cryptic species, commonly detected through karyotypic analyses, which reveals outstanding interespecific variations with relationship to the number and its chromosomal structures. Thus, the present work intends to contribute for the cytogenetic knowledge of marine and brackish fish species, because they peculiar life habits and by lack of cytogenetic data of your genetic aspects. Therefore, cytogenetic studies were developed in a species of Apogonidae (Perciformes), two species of sea catfishes of the family Ariidae (Siluriformes) and brackish fish Paurachenipterus galeatus (Siluriformes, Auchenipteridae), through C banding, Ag-NOR, use of base-specific flourochromes (DAPI and CMA3), as well as FISH (Fluorescent in situ hybridization) using ribosomal DNA probes 5S and 18S. The present results contribute to a better understanding of the processes of differentiation patterns and chromosome evolution in these groups. The use of other approaches (the morphology and molecular tools) will allow a larger understanding of the genetic and biological diversity of the Brazilian ichthyofauna.

Análise citogenética de células-tronco derivadas do subendotélio da veia umbilical humana

Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

Floração precoce em cana-de-açúcar - um estudo utilizando ferramentas de análise in silico e proteômica

Sugarcane is one of the most important products of the world and Brazil is responsible for 25 % of the world production. One problem of this culture at northeast of Brazil is the early flowering. In our laboratory, it has been made before four subtractive libraries using early and late flowering genotypes in order to identify messages related to the flowering process. In this work, two cDNAs were chosen to make in silico analysis and overexpression constructs. Another approach to understand the flowering process in sugarcane was to use proteomic tools. First, the protocol for protein extraction using apical meristem was set up. After that, these proteins were separated on two bidimensional gels. It was possible to observe some difference for some regions of these gels as well as some proteins that can be found in all conditions. The next step, spots will be isolated and sequence on MS spectrometry in order to understand this physiological process in sugarcane

caracterização in situ e estrutura genética de populações de Gossypium mustelinum

Gossypium mustelinum Miers ex Watt is the only cotton species native from Brazil. It is endemic of the semi-arid region from North-east of the country, where it occur near from resilient water sources. The threats to the in situ conservation of the populations are caused by human interference in its habitat, mainly by excessive cattle graze and deforestation. Establish efficient strategies of in situ conservation depend on the accomplishment of a diagnosis of how the specie is found in its natural environment, and the knowledge about the genetic structure of the populations. The objectives of this work were i) to determine the in situ conditions of two populations present in rivers from basin of Rio Paraguaçu at the Bahia State, ii) to evaluate the structure and genetic variability presented in both populations, iii) to establish in situ and ex situ conservation strategies. It were realized collection in november 2007, when was realized in situ characterization of G. mustelinum. SSR markers were used for analyze 218 genotypes deriving from two populations of the G. mustelinum, localized at Tocó river and the Capivara river. The allelic frequencies, the heterozigosity and the F statics were estimated. All the plants were classified as wild and natives, and there was no evidence of the use the plants or its parts. The populations showed different conservation conditions in situ. Few plantlets were found in sites with excessive cattle feed, an indication that the damages in young plants should be high enough to compromise the renovation of the populations. On the other hand, populations were well preserved when the anthropic damages was low or inexistent. The 14 SSR primer pairs amplified 17 loci with a medium number of 5 alleles per locus (a total of 85 alleles). The high level of endogamy estimated (FIS=0,808) and the low observed heterozygosity (H0=0,093) were indicatives that the populations reproduce mainly by selfing, geitonogamy and crosses between related individuals. The genetic diversity was high (HE=0,482) and the differentiation between the populations was very high (FST=0,328). At least two sites from both populations of G. mustelinum must be preserved to achieve suitable in situ conservation. Actions that preserve the gallery forest and keep the cattle away should implemented, and could be as simple as erecting a fence. It is not possible anticipated if the in situ preservation will be possible. Therefore collections and ex situ preservation of representative specimens are essential to conserve the genetic diversity of native G. mustelinum

Estudos ecogenômicos e bioprospectivos de Shewanella ssp

Bacteria trom Shewanella and Geobacter ganera are the most studied iron-reducing microorganisms particularly due to their electron transport systems and contribution to some industrial and environmental problems, including steel corrosion, bioenergy and bioremediation of petroleum-impacted sites. The present study was focused in two ways: the first is an in silico comparative ecogenomic study of Shewanella spp. with sequenced genomes, and the second is an experimental metagenomic work to detect iron-reducing Shewanella through PCR-DGGE of a metabolic gene. The in silico study resulted in positive correIation between copy number of 16S rDNA and genome size in Shewanella spp., with clusters of rrn near lhe origin of replication. This way, the genus is inferred as opportunist. There are no compact genomes and their sequences length varied, ranging from 4306142 nt in S. amazonensis SB2B to 5935403 nt in S. woodyi ATCC 51908, without correIation to temperature range characteristic of each specie. Intragenomic 16S rDNA sequences possess little divergence, but reasonable to resuIt in different phyIogenetic trees, depending on the sequence that is chosen to compare. For moIecuIar detection of iron-reducing Shewanella, it is proposed the mtrB gene as new biomarker. because it codes to a fundamental protein at Fe (III)-reduction. The specific primers were designed and evaluated in silico and resulted in a fragment of 360 pb. In the second study, these primers were tested in a genomic sample from S. oneidensis MR-1, amplifying the expected region. After this successfuI resuIt, the primer set was used as a tool to assess the iron-reducing communities of ShewaneIla genus under an environmental stress, i.e. crude oil contamination in mangrove sediment in Rio Grande do Norte State (Brazil). The primers presented high specificity and the reactions performed resulted in one single band of ampIification in the metagenomic samples. The fingerprinting obtained at DGGE reveaIed temporal variation of Shewanella spp. in analyzed samples. The resuIts presented show the detection of a biotechnological important group of microorganisms, the iron-reducing Shewanella spp. using a metabolic gane as target. It is concluded there are eight or more 16S rDNA sequences in Shewanella genus, with little divergence among them that affects the phylogeny; the pair of primers designed to ampIify mtrB sequences is a viable alternative to detect iron-reducing ShewanelIa in metagenomic approaches; such bacteria are present in the mangrove sediment anaIyzed, with temporal variations in the samples. This is the first experimental study that screened the iron-reducing Shewanella genus in a metagenomic experiment of mangrove sediments subjected to oil contamination through a key metabolic gene

Análise das mudanças no perfil protéico durante o estresse oxidativo in vivo e atuação da MutY-Glicosilase em respostas celulares

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Avaliação de pastagens tropicais em sistemas de produção de ovinos de corte no nordeste brasileiro

He aim of this study was to evaluate the structural characters, herbage accumulation, nutritive value and performance of sheep in different tropical pastures. The treatments were two cultivars and two of the genera Panicum Brachiaria pastures under intermittent stocking and variable stocking rate, in the rainy season. We evaluated the masses and the components of herbage pre grazing in two layers, and after grazing. Chemical analyzes were made of the stems and leaf blades pre grazing in two layers. We used 48 male sheep and whole for the assessment of individual weight gain and area, and anestrous females to adjust the stocking rate. In the cv. Massai showed the highest herbage mass, leaf blades and dead material, and the largest volume density and leaf blade: stem pre grazing. There was no difference among cultivars for the percentage of leaf blade (PLB) in both strata, but the higher the PLB was higher than the bottom. The highest percentage of dead material (PDM) was observed in cvs. Massai and Marandu in the two strata. In cvs. Massai and Piatã were observed lower levels of crude protein in stem and leaf. In stratum 0-25 cm lower nutritional value was observed in the stem in the leaf blades did not grant the nutritional value among the strata. Herbage mass, leaf blade, PLB and proportion of stem in the residue of the Massai pastures were higher than cv. Aruana. There was no difference for efficiencies in harvest leaf and stem between the cultivars. The cv. Massai got the higher accumulation of leaf per cycle per day. Animals kept in grass swards Aruana had the highest average daily gain. The higher stocking rates and earnings per area were observed in grass swards and Marandu Massai. The cultivars are suitable for sheep meat production in the rainy season

Avaliação de métodos de análises para determinação da contagem de células somáticas no leite cru, mantido em tanque de resfriamento

We analyzed the quality of raw milk from eight dairy farms in Rio Grande do Norte stored in a cooling tank , in order to evaluate methods for determining somatic cell counts (SCC). The Somaticell® kit and a portable Direct Cell Counter (DCC) were compared with each other and with the MilkoScanTM FT+ (FOSS Denmark), which uses Fourier Transform Infrared (FTIR) spectroscopy). Direct cell counter data were processed for somatic cell scores (log-transformed somatic cell count) and analyzed with the SAS®, statistical package , Statistical Analysis System, (SAS, INSTITUTE, 1998). Comparison of means and correlation of somatic cell scores were conducted using Pearson s correlation coefficient and the Tukey Test at 1 %. No significant difference was observed for comparison of means. The correlation between somatic cell scores was significant, that is, 0.907 and 0.876 between the MilkoScanTM FT+ and the Somaticell® kit and Direct Cell Count (DCC) respectively, and 0.943 between the Somaticell® kit and Direct Cell Count (DCC). The methods can be recommended for monitoring the quality of raw milk kept in a cooling tank in the production unit

Marcadores fenotípicos para caracterização de caprinos com diferentes níveis de resistência às endoparasitoses gastrintestinais

The aim of this study was to evaluate and characterize phenotypically goats with different levels of resistance to gastrointestinal nematodes. For a period of 93 days, 60 F2 goats originated from ½ Saanen and ½ Anglo- nubian animals were kept in the same area of pasture. Every seven days, feces and blood were collected for eggs per gram counts of feces (EPG) and cultures of feces and to determinate the number of eosinophils, packed cell volume and total plasma protein, respectively. On the same day, the animals were weighed and submitted to body score condition and FAMACHA method to worm control. Based on the average of EPG, the twelve animals with the highest average (susceptible group) and the twelve animals with the lowest average of EPG (resistant group) were selected, slaughtered and necropsied to recovery, counting andparasites identification. The resistant animals present lower EPG mean (P <0.0001) and 4.7 folder less parasites than susceptible animals. The resistant group presented higher mean packed cell volume (26.48%) and total plasma protein (6.24 g / dl) than susceptible one (24,04% e 5,82g/dl, respectively). The average number of eosinophils was similar in both groups The Haemonchus sp. was the most prevalent in the culture of feces, followed by Trischostrongylus sp. and Oesophagostomum sp.. The counting of nematodes in the abomasum of susceptible group was higher than in resistant one. The species identified were H. contortus in abomasums and T. colubriformis in small intestine. It can be concluded that EPG, packed cell volume and total plasma protein were useful phenotypic markers to identify animals as resistant and susceptible to gastrointestinal nematodes infections

Fatores de influência na produção de leite bovino sobre os níveis da contagem bacteriana total

The objective of this study was to evaluate the influence of milking procedures on the levels of total bacterial count (TBC) in bovine milk. In the first study the influences of procedures for hygienic milking, cleaning of milking equipment and milk cooling tanks on the TBC levels were evaluated. Four bulk samples of milk were collected from each tank in eight properties for TBC analysis, employing the flow cytometry method. A questionnaire was applied in each property to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology for analysis of longitudinal data was considered, focusing on random effects models. The results showed that the handling procedures for milking and the cleanliness of the cooling tank contributed to a further reduction in the levels of TBC raw milk cooling tanks. The second study aimed to describe the percentage of the properties that comply with the Normative Instruction Nº 51 (Brazil s IN 51) with regard to total bacterial count (TBC) in bovine milk. The study was conducted from January 2010 to July 2011. Milk samples were collected from the eight properties selected for TBC analysis by the flow cytometry method. Again, on each property a questionnaire was applied to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology of marginal models based on Generalized Estimate Equations (GEEs) was followed in the statistical analysis. The results showed that the handling procedures of the milking and the cleanliness of the cooling tanks contributed to a considerable percentage of the properties that reached the limits of TBC established by IN 51

Avaliação dos impactos ambientais e sociais gerados pela utilização do kit Embrapa de ordenha manual na produção de leite de cabra

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Caracterização dos sistemas de produção de leite bovino na microrregião Seridó do estado do Rio Grande do Norte

The objective was to contribute to a reading of the characteristics and diversity of production systems from bovine milk in the microregion Seridó of Rio Grande do Norte, discuss the social, economic, environmental and husbandry-related primary activity of milk production. We randomly selected 28 agricultural establishments that performed the activity of dairy bovine culture with subsequent applicat ion of a structured questionnaire during September and October 2011. Data were analyzed with application of measures of descriptive analysis and determination of the index rural development (IRD). The results showed that 53.57% of the interviewees were owners of the land, the median area of the properties amounted to 135 hectares, the median number of animals in the herd was 51 head, with minimum 11 and maximum of 350 heads establishments in the sample, 85.72% of establishments had maximum 23 cows in lactation, 100% performed manual milking with suckling calves, average productivity of 3.91 liters / cow / day, 92.86% of the interviewees did not produce silage and / or hay, 64.29% had no access to technical assistance, the average age of interviewees was 51 ± 10.85 years, 78.57% had only elementary education. The average of the IRD computed in the test sample amounted to 0.43 ± 0.11 on a scale 0-1. The sites were grouped into 04 classes, based on your IRD. The average IRD in each dimension was 0.43, 0.55 and 0.34, respectively for economic / husbandry, social and environmental. The determination of the rate of rural development - IDR found for the sample could add establishments hierarchically. It is necessary that the inclusion of best practices in herd management, bookkeeping zootechnical, technical assistance and reorganization practices in land and environmental preservation

Caracterização genética de vacas leiteiras por meio de marcadores moleculares e suas implicações na composição e qualidade do leite

The objective of this study was to identify DNA polymorphisms at the genes leptin, β-lactoglobulin and pituitary-specific transcription factor in three genetic groups of Holstein x Guzerat dairy cows and investigate the relationship between their genotypes and the composition and quality of milk of dairy cows. Samples were collected in August 2009, being 113 blood samples from lactating crossbred cows and 58 milk samples. For analysis of DNA polymorphisms blood samples were collected, analyzed later in the Genetic Laboratory affiliated to the Zootechny Institute of São Paulo and individual milk samples were collected according to standards established by the laboratory of Management Program of Northeast Dairy Herds (PROGEN), at Federal Rural University of Pernambuco (UFRPE) for analysis of milk composition and quality. The characterization of genotypes was performed by PCR-RFLP, for which were designed specific primers for each studied gene and restriction enzymes Kpn2I, HaeIII and HinfI that cut the DNA of the following genes: leptin, β-lactoglobulin and a PIT, respectively. The leptin estimate genotypic frequence were CC 0.112, TT 0.225 and CT 0.661, for β-lactoglobulin were AA 0.136, AB 0.323 and BB 0.539, and for PIT were ++ 0.655, -- 0.311 and +- 0.032. The results show that the population is in Hardy-Weinberg disequilibrium for leptin, β-lactoglobulin and a PIT due to excess of heterozygotes in the population, however, as these genes are associated with the milk production it is considered that the animals have genetic potential for milk production in the Brazilian semi-arid conditions. Through the characterization of the studied herd there were not found implications of the polymorphism of leptin, β-lactoglobulin and PIT in the composition and quality of milk from cows in the different genetic groups 1/2, 3/4 and 7/8 Holstein x Guzerat. Key words: β-lactoglobulin, crossbred cows, leptin, PCR-RFLP, PIT1, semi-arid.

Suplementacao de zinco orgânico na racão de frangos de corte alojados em cama nova ou reutilizada

Abstract: Several factors can affect the development of the broiler, among them we can highlight nutrition and management. In the context nutritional, mineral supplementation is a necessary practice because, in general, the diets did not contain these elements in sufficient quantity to meet the needs of poultry. Zinc is a trace mineral essential to life, participating in several important functions in the body. Generally zinc is added to diets of birds in inorganic forms (oxides, carbonates or sufatos), however in its organic form or chelated presents more bioavailable. The objective of this study was to evaluate the effect of levels of organic zinc (ZnO) in the diet of broilers from 1 to 42 days, housed in new or reused litter. The experiment was conducted in the poultry sector of the Special Unit for Agricultural Sciences EAJ / UFRN. 576 chicks were used 1 day of commercial strain Cobb, distributed in a completely randomized in a 4x2 factorial arrangement with four levels of ZnO 0, 40, 80 and 120 ppm and two environments, new bed (COn) and reused litter (CRE) resulting in eight treatments with six replications of 12 birds. In the pre-initial responses were linearly increasing levels of ZnO on feed intake and quadratic effect on body weight and weight gain. The levels of 72.41 and 70.05 ppm of ZnO in the diet of chicks improved body weight and weight gain, respectively. There was interaction between ZnO and the type of bedding used. The ZnO did not affect broiler performance in the growing phase. There was an interaction between levels of ZnO and type of bed used. The levels of 61.50 and 85.30 ppm organic zinc improves immunity and increases the deposition of zinc in tibia of broilers at 42 days, respectively. ZnO also increases the resistance of the skin of broilers at 42 days of age. Using Cre improves performance of broilers from 1 to 42 days old

Características da carcaça e qualidade da carne de ovinos terminados em pastagens cultivadas

The objective of this study was to evaluate the quality of housing and the physical and chemical characteristics of meat from sheep raised on pasture Brachiaria brizantha and Panicum maximum. The experiment was conducted in the physical area of the Study Group on Forage (GEFOR), located in the Academic Unit Specialized in Agricultural Sciences - Federal University of Rio Grande do Norte - UFRN in Macaíba, RN, Brazil. We used 32 lambs SPRD, obtained from herds in the state, with liveweight (LW) of 24.5 kg were assigned randomly to four treatments consisting of tropical grasses, two cultivars of Brachiaria brizantha, Marandu and Piatã, and two of Panicum maximum, Aruana and Massai. The experimental area was 2.88 ha, divided into 4 paddocks of 0.72 ha, where each picket consisted of a farm and was divided into six plots of 0.12 ha, where the animals remained under rotational grazing. The period of adaptation to the pickets was seven days. At the beginning of the experiment the animals were weighed, identified with plastic earrings and necklaces colored according to the treatment, and treated against. The lambs were loose in the paddock at 8 am and collected at 16 hours, which returned to collective pens. During the time of grazing animals had free access to mineral supplement with monensin Ovinofós ® and water. Before entering the paddocks of pasture were sampled to characterize the chemical composition. Every seven days occurred at weighing, with fasting, to monitor the weight development. Cultivars Marandu, Aruana, Piatã and Massai were grazed for 133, 129, 143 and 142 days, respectively, until the lambs reach slaughter weight. Arriving at 32 kg lambs were evaluated subjectively for body condition score by, passed through fasting period, diet and water for 16 hours were slaughtered. Measurements were made in the inner and outer casings in addition to subjective evaluations regarding muscling, finish and quantity of pelvic-renal fat, then each was divided longitudinally into two half-carcases and cuts were made in the commercial left half, and after heavy calculated their income. Between the 12th and 13th thoracic vertebrae, was performed a cut to expose the cross section of the Longissimus dorsi, which was drawn on the rib eye area (REA) in transparent film. Fat thickness and extent of AOL GR were determined using a caliper. A tissue composition was determined by dissection of the legs. Analyzes were performed physical (color, cooking loss and shear force) and chemical composition of meat (moisture, ash, protein and lipids) in Longissimus dorsi muscle. Grazing tropical grass Brachiaria brizantha cvs. Marandu and Piatã and Panicum maximum cvs. Aruana and Massai can be used for lambs SRPD in the rainy season, because not alter the physico-chemical and chemical composition of meat