Early Development and Putative Primordial Germ Cells Characterization in Dogs

Contents Previously, three distinct populations of putative primordial germ cells (PGCs), namely gonocytes, intermediate cells and pre-spermatogonia, have been described in the human foetal testis. According to our knowledge, these PGCs have not been studied in any other species. The aim of our study was to identify similar PGC populations in canine embryos. First, we develop a protocol for canine embryo isolation. Following our protocol, 15 canine embryos at 21-25 days of pregnancy were isolated by ovaryhysterectomy surgery. Our data indicate that dramatic changes occur in canine embryo development and PGCs specification between 21 to 25 days of gestation. At that moment, only two PGC populations with distinct morphology can be identified by histological analyses. Cell population 1 presented round nuclei with prominent nucleolus and a high nuclear to cytoplasm ratio, showing gonocyte morphology. Cell population 2 was often localized at the periphery of the testicular cords and presented typical features of PGC. Both germ cell populations were positively immunostained with anti-human OCT-4 antibody. However, at day 25, all cells of population 1 reacted positively with OCT-4, whereas in population 2, fewer cells were positive for this marker. These two PGCs populations present morphological features similar to gonocytes and intermediate cells from human foetal testis. It is expected that a population of pre-spermatogonia would be observed at later stages of canine foetus development. We also showed that anti-human OCT-4 antibody can be useful to identify canine PGC in vivo.

Fatty acid composition of intramuscular fat from Nellore steers fed dry or high moisture corn and calcium salts of fatty acids

The purpose of this work was to evaluate the fatty acid composition of the Longissimus muscle from carcasses of Nellore steers fed diets with calcium salts of fatty acids (CSFA) and high moisture corn. Forty eight steers were fed during 70 days four diets containing dry corn (DC), high moisture corn (HM). dry corn plus CSFA (DC-CSFA) or high moisture corn plus CSFA (HM-CSFA). Fatty acid composition of the Longissimus muscle was determined by gas chromatography. Corn type had no effect on the ether extract percentage and in the content of the majority of the fatty acids, although steers fed HMC showed higher levels of polyunsaturated fatty acids and polyunsaturated/saturated ratio. Feeding CSFA increased ether extract percentage but had no effect on total of saturated, unsaturated and saturated: unsaturated ratio. Both high moisture corn and calcium salts of fatty acids increased CIA (cis9, trans11) and total CIA concentrations in intramuscular fat (C) 2008 Elsevier B.V. All rights reserved.

PHYSICAL AND CHEMICAL CHARACTERISTICS OF FROZEN GROUND BEEF AND AGED BEEF MEAT FROM BOS INDICUS STEERS SUPPLEMENTED WITH alpha-TOCOPHEROL ACETATE

The objective of this study was to evaluate the effects of diet supplementation with vitamin E on the physical and chemical characteristics of ground, frozen and stored or aged Quadriceps femoris (QF) and Longissimus dorsi (LD) muscles from Nellore steers fed high concentrate diets. Muscles were obtained from 24 animals that were 30 months old with a mean live weight of 279 kg. Half of the animals received daily doses of 1,000 mg of alpha-tocopherol acetate (VIT E) per head per day that was added to 100 g of corn meal. The other half received 100 g of corn meal without the antioxidant. Twenty-four hours after slaughtering, QF samples from each animal were ground, frozen and stored for up to 6 months. In addition, 4 samples from the LD of each animal were vacuum packed individually and kept for 21 days. All samples were analyzed to determine the pH, color and water-holding-capacity. The VIT E supplementation improved only the water loss characteristics of frozen ground QF and did not have any positive effect on the physical-chemical characteristics of the aged LD.

Relative populations and toxin production by Aspergillus flavus and Fusarium verticillioides in artificially inoculated corn at various stages of development under field conditions

The presence, development and production of mycotoxins by Aspergillus flavus and Fusarium verticillioides were studied in corn ears under field conditions after artificial contamination of corn silks. The planted area was divided into five treatments: T1, inoculated with A.flavus solution containing 1 x 10(8) spores, ears covered; T2, inoculated with F. verticillioides solution containing 1 X 10(8) spores, ears covered; T3, inoculated with E verticillioides plus A. flavus solution containing 1 x 10(8) spores of each, ears covered; T4, sprayed with sterile phosphate-buffered saline, ears covered; TS, non-sprayed silks, uncovered ears. Soil and air samples were also collected and analysed for the occurrence of fungi. Water activity, relative air humidity, rainfall and temperature were determined to assess the correlation between abiotic factors and the presence of fungi in the samples. Contamination with the inoculated fungus predominated in T1 and T2. In the other treatments, F. verticillioides was the most frequently isolated contaminant irrespective of treatment. Considering the production of mycotoxins, a positive relation between the production of fumonisins B-1 and B-2 and the frequency of F. verticillioides was statistically verified in all treatments. (C) 2007 Society of Chemical Industry.

Influence of macro- and micronutrient fertilization on fungal contamination and fumonisin production in corn grains

The aim of this study was to investigate the effects of nutrients (nitrogen, zinc and boron) on fungal growth and fumonisins production in corn samples obtained at the beginning of grain formation and at harvest. Three nitrogen doses were applied to the corn plants through soil in combination with three zinc doses and two boron doses during sowing. Mycological analysis of grains, using Dichloran Rose-Bengal Chloramphenicol Agar, collected at the beginning of formation demonstrated a fungal population predominantly of yeasts. Analysis of freshly harvested corn revealed a higher frequency of Penicillium spp. (72%) and F verticillioides (27%). High Performance Liquid Chromatography analysis revealed that 100% of grains were contaminated with fumonisins B, at levels ranging from 0.3 to 24.3 mg/kg and 93% contaminated with fumonisin B(2) at levels ranging from 0.05 to 5.42 mg/kg. Nitrogen (50 kg/ha) in combination with boron (0.5 kg/ha) resulted in an increased fumonisin B2 production. (c) 2007 Elsevier Ltd. All rights reserved.

Molecular characterization and fumonisin production by Fusarium verticillioides isolated from corn grains of different geographic origins in Brazil

Gibberella moniliformis is most commonly associated with maize worldwide and produces high levels of fumonisins, some of the most agriculturally important mycotoxins. Studies demonstrate that molecular methods can be helpful for a rapid identification of Fusarium species and their levels of toxin production. The purpose of this research was to apply molecular methods (AFLP, TEF-1 alpha partial gene sequencing and PCR based on MAT alleles) for the identification of Fusarium species isolated from Brazilian corn and to verify if real time RT-PCR technique based on FUM1 and FUM19 genes is appropriated to estimate fumonisins B(1) and B(2) production levels. Among the isolated strains, 96 were identified as Fusarium verricillioides, and four as other Fusarium species. Concordant phylogenies were obtained by AFLP and TEF-1 alpha sequencing, permitting the classification of the different species into distinct clades. Concerning MAT alleles, 70% of the F. verricillioides isolates carried the MAT-1 and 30% MAT-2. A significant correlation was observed between the expression of the genes and toxin production r=0.95 and r=0.79 (correlation of FUM1 with FB(1) and FB(2), respectively, P < 0.0001): r=0.93 and r =0.78 (correlation of FUM19 with FB(1) and FB(2). respectively, P < 0.0001). Molecular methods used in this study were found to be useful for the rapid identification of Fusarium species. The high and significant correlation between FUM1 and FUM19 expression and fumonisins production suggests that real time RT-PCR is suitable for studies considering the influence of abiotic and biotic factors on expression of these genes. This is the first report concerning the expression of fumonisin biosynthetic genes in Fusarium strains isolated from Brazilian agricultural commodity. (c) 2010 Elsevier B.V. All rights reserved.

The doodle of a finitely determined map germ from R(2) to R(3)

Let f : U subset of R(2) -> R(3) be a representative of a finitely determined map germ f : (R(2), 0) -> (R(3), 0). Consider the curve obtained as the intersection of the image of the mapping f with a sufficiently small sphere s(epsilon)(2) centered at the origin in R(3), call this curve the associated doodle of the map germ f. For a large class of map germs the associated doodle has many transversal self-intersections. The topological classification of such map germs is considered from the point of view of the associated doodles. (C) 2009 Elsevier Inc. All rights reserved.

Humming as a mean of communicating during meal time situations : A Single Case study involving a women with severe dementia and her caregiver

Objective: ‘Music Therapeutic Caregiving’, when caregivers sing for or together with persons with dementia during morning care situations, has been shown to increase verbal and nonverbal communication between persons with dementia and their caregivers, as well as enhance positive and decrease negative emotions in persons with dementia. No studies about singing during mealtimes have been conducted, and this pilot project was designed to elucidate this. However, since previous studies have shown that there is a risk that persons with dementia will start to sing along with the caregiver, the caregiver in this study hummed such that the person with dementia did not sing instead of eat. The aim of this pilot project was threefold: to describe expressed emotions in a woman with severe dementia, and describe communication between her and her caregivers without and with the caregiver humming. The aim was also to measure food and liquid intake without and with humming. Method: The study was constructed as a Single Case ABA design in which the ordinary mealtime constituted a baseline which comprised a woman with severe dementia being fed by her caregivers in the usual way. The intervention included the same woman being fed by the same caregiver who hummed while feeding her. Data comprised video observations that were collected once per week over 5 consecutive weeks. The Verbal and Nonverbal Interaction Scale and Observed Emotion Rating Scale were used to analyze the recorded interactions. Results: A slightly positive influence of communication was shown for the woman with dementia, as well as for the caregiver. Further, the women with dementia showed a slight increase in expressions of positive emotions, and she ate more during the intervention. Conclusion: Based on this pilot study no general conclusions can be drawn. It can be concluded, however, that humming while feeding persons with dementia might slightly enhance communication, and positive expressed emotions in persons with dementia. To confirm this, more studies on group levels are needed. Because previous studies have found that caregiver singing during caring situations influences persons with dementia positively it might be desirable to test the same during mealtime.

Desempenho e eficiência biológica de bubalinos de três grupos genéticos terminados em confinamento e abatidos em diferentes estádios de maturidade

Neste trabalho objetivou-se estudar o desempenho e a eficiência biológica de 48 bubalinos (16 Murrah - MUR, 16 Jafarabadi - JAF e 16 Mediterrâneo - MED) com média de 18 meses de idade e de peso vivo inicial de 330 kg terminados em confinamento. Quatro animais de cada grupo genético foram designados por sorteio para o abate inicial e serviram de referência para o estudo dos pesos iniciais de corpo vazio e de carcaça. Os 12 animais restantes de cada grupo genético foram divididos aleatoriamente em três subgrupos de quatro animais e submetidos aos seguintes tratamentos: Maturidade 1 (400 kg PV ao abate); Maturidade 2 (450 kg PV ao abate); Maturidade 3 (500 kg PV ao abate). Foi fornecida uma ração única, ad libitum, para todos os animais, composta por 50% de feno de coastcross na MS e 50% de concentrado à base de fubá de milho, uréia e farelo de soja, devidamente suplementado com minerais. Após os abates pré-fixados, determinou-se o peso corporal vazio (PCVZ) dos animais, por meio do somatório das partes integrantes do corpo. Os consumos médios diários de MS e MO não diferiram entre grupos genéticos, embora animais JAF tenham apresentado os maiores valores numéricos. Animais MUR, JAF e MED não diferiram quanto aos ganhos de peso corporal vazio e de carcaça. Não houve diferenças quanto à eficiência biológica (ganho de peso corporal vazio e ganho de carcaça por unidade de energia metabolizável ingerida) entre os grupos genéticos, embora animais Murrah tenham apresentado os maiores valores. Bubalinos abatidos aos 400 e 450 kg apresentaram melhor eficiência de ganho de peso corporal vazio e ganho de carcaça que os abatidos aos 500 kg.

Germ Cell Transplantation in Felids: A Potential Approach to Preserving Endangered Species

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Farinha de resíduos da indústria de filetagem de tilápias como fonte de proteína e minerais para alevinos de tilápia-do-nilo (Oreochromis niloticus)

Neste trabalho, objetivou-se avaliar a utilização da farinha de resíduos da indústria de filetagem de tilápias (FR) como fonte de proteína e de minerais em rações práticas na alimentação de alevinos de tilápia do Nilo. Foram utilizados 120 alevinos de tilápias do Nilo com peso e comprimento iniciais de 0,58 ± 0,05 g e 3,49 ± 0,09 cm, respectivamente, distribuídos em 24 aquários com capacidade para 30 L, em um delineamento inteiramente casualisado, com quatro tratamentos e seis repetições. Os peixes foram alimentados com rações contendo 30% de proteína digestível e 3.000 kcal de energia digestível/kg, de acordo com os seguintes tratamentos: CO - ração à base de milho e farelo de soja, sem suplementação de fósforo; FB - ração à base de milho e farelo de soja, com fosfato bicálcico; FB + FR - ração à base de milho e farelo de soja, suplementada com fosfato bicálcico (50%) e farinha de resíduos (50%); FT - ração à base de milho e farelo de soja, suplementada com farinha de resíduos. Ao final do experimento, os melhores resultados de desempenho foram observados nos animais que receberam suplementação de P. Quanto às características de carcaça, os animais que receberam a ração CO apresentaram maior teor de gordura corporal e menores teores de cinzas, Ca e P. A FR pode ser utilizada em rações para alevinos de tilápia do Nilo como fonte de P, sem prejuízos no desempenho e na composição corporal.

Corn (Zea mays L.) cultivars tolerance to nicosulfuron

The effects of nicosulfuron on morphological, yield and growth characteristics and the histological modifications in the leaf blade were evaluated for eight corn cultivars: landrace popcorn, ESALQ-popcorn, Piranao, DSCC-Architecture, ESALQ VF7, ESALQ VD8, JAB 01F and JAB 02D. The experimental de sign was a randomized complete block with three replications in a factorial arrangement. Plots were four rows wide and 6.00m long with 0.90m spacing between rows and 0.20m between plants. Nicosulfuron was applied at the dose of 160 g a.i.ha(-1), when the plants was 25 cm high and has 5-6 expanded leaves. Nicosulfuron significantly reduced plant height at silking and the Final plant stand. There was also a significant reduction for plant height at 47 clays after sowing, for number of leaves, ear position. ear height, grain yield, ear weight and total number of ears due to nicosulfuron. The cultivars JAB 01 and JAB 02 were the least affected by the herbicide with similar yields to those of the control group, showing good tolerance to nicosulfuron. The leaves injury symptoms for all the studied cultivars consisted of chlorosis and puckering of the blades from the expanding central leaves of the plant at 7 days after application. Anatomical alterations in the epidermis due to the herbicide consisted of an increase in number and size of the bulliform cells and in the suppression of the epidermal cells in the epidermis and in the nearby of the vascular bundles. Some parenchymatic cells became voluminous and devoid of chloroplasts. SEM examination revealed a loss of bilateral symmetry of the stomata and disappearance of the subsidiary cells from the stomata near the midrib at the adaxial surface.

RAPD marker use for improving resistance to Helicoverpa zea in corn

Helicoverpa zea is responsible for great losses to the corn, Zen mays L., crops final productivity, and the best way to control it is by improving genetic resistance. In collaboration with corn improvement and increasing resistance to insects through molecular marker assisted selection, this work had as an objective the selection of resistant (RP) and susceptible progenies (SP) to H. zea based on the RAPD technique. Molecular markers were Found, among the resistant progenies and it is suggested that linkage of these within the Zapalote Chico corn race, be used to extract resistance genes from this race as a donor. The progenies were selected from a population of half-sibs exhibiting a broader genetic base (FCAVJ-VF14). After DNA extraction, two sample bulks were formed; one made up of the six most resistant plants, the other of the six least resistant plants. Eighty-six primers were tested for PCR reactions with the resistant and susceptible bulks and analyzed on agarose electrophoresis for the detection of RAPD band polymorphism. The results of the banding patterns and similarity values indicated a nucleotide sequence amplified by the primer OPC-2 as a possible molecular marker for the identification of resistant progenies and a homology region between them and the Zapalote Chico corn race.

Crescimento e esporulação de duas espécies de arthrobotrys corda em diferentes meios de cultura e dois ambientes

The researches on the biological control of nematodes with nematophagous fungi has been intensified in recent years. The knowledge of the ecological conditions for the growth and sporulation of these fungi is a prerequisite for attainment of pure cultures needed to attend the demand for formulation of these organisms. With the objective to evaluate the micelial growth and sporulation of Arthrobotrys musiformis and A. oligospora in two environments (B.O.D at 25 +/- 1 degrees C and the environment of the Laboratory), 20 cultures media prepared with common materials found in the communities and industrialized media such as mycological agar, PDA and CMA were evaluated. The media were tested in Petri dishes, being the micelial growth of the fungi evaluated daily, during six days. The measured sporulation at the end of the experiment was done by estimation of the number of conidia/Petri dish. The experiment was carried out in a random design following a factorial arrangement of 20 x 2 x 2, corresponding to 20 media, two fungi and two environments, with five replicates. The variance analysis of the data evidenced significant statistical difference by the F Test, at 1% probability, among media x fungi x environment interaction. Fifty percent of the tested media provided the adequate micelial growth of A. musiformis and there was no statistical difference among them, namely: cassava meal (FM), sweet starch (PD), "corn meal agar" (CMA), oat in fine flakes (AFF), agar-water + dextrose (AA+D), mycological agar (AM), potato dextrose agar (BDA), meal of maize (FMI), flour of wheat (FT) and wheat for kibble (TK). In relation to A. oligospora, 75% of the tested media promoted the maximum growth of the fungus, which are: AFF, AM, FM, PD, CMA, AA+D, BDA, FT, TK, the water from the decoction of rice (AAZ), rice in grains (AZG), triturated rice (AZT), thread flour (FR), oats flour (FA), oats in thick flakes (AFG) and flour of maize (FU). In relation to the sporulation the media that had better role for A. musiformis, in decreasing order, were: FR, TK, AFG, BDA, FA, AFF, AM, FMI, AZT and FM, varying between 1,01 x 10(6) and 1,4 x 10(4) conidia/Petri dish. For the A. oligospora sporulation, the CMA medium provided the maximum level with an estimated average of 5,7 x 10(6) conidia/Petri dish. In the general, the best media for the micelial growth and sporulation of A. musiformis had also been the best for A. oligospora. However, some that had been the best for the A. oligospora did not had been efficient for the micelial growth or the sporulation of A. musiformis, indicating that the isolate of A. musiformis in case is more demanding than that A. oligospora one. The evidences from the study indicate that, in Jaboticabal, São Paulo state, the growth and the sporulation of these fungi do not demand special chambers. Some adaptations of an environment at the laboratory, enough to obliterate the light are sufficient.

Anaerobic biodigestion of pigs feces in the initial, growing and finishing stages fed with diets formulated with corn or sorghum

Objetivou-se avaliar a produção e a qualidade do biogás e do biofertilizante obtidos em biodigestores abastecidos com dejetos de suínos, nas fases inicial, crescimento e terminação, alimentados com dietas formuladas à base de milho ou sorgo. Foram utilizados 20 biodigestores de bancada, com tempo de retenção hidráulica de 30 dias, e cargas diárias que continham 4,0 a 6,0% de sólidos totais (ST) e 3,6 a 5,2% de sólidos voláteis (SV). Nos efluentes dos biodigestores, foram verificados teores médios de ST variando de 1,6 a 2,0% e de SV entre 1,2 e 1,6%. As reduções médias de ST foram de 57,7 a 64,7% e de SV de 61,7 a 69,0%, sendo que houve diferença somente na fase de terminação, na qual as maiores reduções médias foram para biodigestores abastecidos com dejetos de animais alimentados com dietas à base de milho. Nos biodigestores abastecidos com dejetos de animais em fase inicial e de crescimento, alimentados com dietas formuladas à base de milho, foram verificadas maiores produções médias de biogás e os maiores potenciais médios de produção de biogás. Os potenciais médios obtidos foram 0,033; 0,181; 0,685; 0,788 e 1,132 m³ por kg de afluente, estrume, ST adicionados, SV adicionados e SV reduzidos, respectivamente. Não foram verificadas diferenças quanto ao teor médio de metano no biogás entre dietas e fases. As concentrações médias dos nutrientes N, P, K, Ca, Mg, Na, Fe, Mn, Zn e Cu no afluente e efluente dos biodigestores variaram entre as dietas e fases.