Expression levels of mRNA-encoding PDGF receptors in goat ovaries and the influence of PDGF on the in vitro development of caprine pre-antral follicles

The aims of this study were to investigate the expression levels of mRNA for platelet-derived growth factor (PDGF) receptors (PDGFR-α and -β) in caprine follicles at different developmental stages and to evaluate the influence of PDGF on the in vitro development of pre-antral follicles. For this, goat primordial, primary and secondary follicles, as well as small (1-3 mm) and large (3-6 mm) antral follicles, were obtained, and PDGFR-α and -β mRNA levels were quantified by real-time PCR. Furthermore, pre-antral follicles (≥ 200 μm) were isolated from goat ovaries and cultured for 18 days in α- minimum essential medium supplemented with PDGF at 50 or 100 ng/ml, containing or not FSH. Real-time PCR showed highest PDGFR-α mRNA levels in secondary follicles, while PDGFR-β mRNA levels were highest in primary follicles onwards. Both receptors showed higher mRNA levels in granulosa/theca cells from small and large antral follicles than in their corresponding cumulus-oocyte complexes. In culture, the percentage of antrum formation was significantly higher in 100 ng/ml PDGF compared with the same PDGF concentration associated with FSH. After 18 days, PDGF in both concentrations associated with FSH promoted follicular growth significantly higher than the control. Moreover, the addition of FSH to 50 ng/ml PDGF positively influenced the follicular growth when compared with the same PDGF concentration in the absence of FSH. In conclusion, PDGF is important for early goat folliculogenesis, because the presence of PDGFR-α and -β mRNA was detected in all follicular categories, and PDGF associated with FSH stimulated the growth of goat pre-antral follicles isolated and cultured in vitro.     

Proteomic analyses of Ehrlichia ruminantium highlight differential expression of MAP1-family proteins

The Rickettsiales Ehrlichia ruminantium (ER) is the causative agent of heartwater, a fatal tick-borne disease of livestock in sub-Saharan Africa and in the Caribbean, posing strong economical constraints to livestock production. In an attempt to identify the most prominent proteins expressed by this bacterium, especially those encoded by the major antigenic protein 1 (map1) multigene family, a proteome map of ER cultivated in endothelial cells was constructed by using two dimensional gel electrophoresis combined with mass spectrometry. Among the sixty-four spots detected, we could identify only four proteins from the MAP1-family; the other proteins detected were mainly related to energy, amino acid and general metabolism (26%), to protein turnover, chaperones and survival (21%) and to information processes (14%) or classified as hypothetical proteins (23%). Additional studies on MAP1-family protein using immunochemical labeling also revealed that these proteins are differentially expressed along the bacterium life cycle, presenting different structural organization. Interestingly, when infectious elementary bodies (EBs) are released from host cells, MAP1 appears to be organized in SDS and heat-resistant dimers and trimers stabilized by disulfide bridges. Overall, the results presented herein not only reveal the first partial proteome map of ER but provide new insights on the expression ER MAP1-family proteins in host endothelial cells.     

Ovarian Follicular Dynamics of five-eighths Girolando Cows

The objective of this study was to examine the follicular dynamics of five-eighths Girolando cows by observing the number of follicular development waves, days of emergence of those waves, diameters of the dominant and largest subordinate follicles and the processes of follicular selection and dominance. Ovarian follicle dynamics were monitored for 24 oestrous cycles in 12 cows, of 4 to 10 years of age, presenting regular oestrous cycles and with body scores between 3 and 4. Ovaries were observed daily for two consecutive oestrous cycles and follicles were measured with ultrasonographic equipment. The dominant follicle was considered to be that which presented a diameter ≥ 10 mm for three consecutive days and on the day of wave emergence, when a group of follicles measuring 3–5 mm in diameter would appear. Of the 24 cycles monitored, 62.5% presented two waves of follicular development and 37.5% presented three waves. The cycles presenting two waves had an average duration of 20 days, with the emergence of the waves on days 1 and 9, whereas the cycles presenting three waves had an average duration of 22 days, with emergences on days 1, 10 and 16. For cycles with two waves, both the first and second dominant follicles reached an average size of 13.8 mm, with the first regressing on day 10 and the second ovulating around day 20. For cycles with three waves, the dominant follicles of the first and second waves reached maximum diameters of 11.8 and 12 mm, respectively, with the first regressing on day 11 and the second regressing on day 17. The third dominant follicle reached a maximum diameter of 12.4 mm on day 20, and ovulated on day 22. These results lead to the conclusion that the follicular dynamics of five-eighths Girolando cows are characterized by the presence of two to three waves of follicular growth.     

Heterofucan from Sargassum filipendula induces apoptosis in HeLa cells

Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated l-fucose. Heterofucan SF-1.5v was extracted from the brown seaweed Sargassum filipendula by proteolytic digestion followed by sequential acetone precipitation. This fucan showed antiproliferative activity on Hela cells and induced apoptosis. However, SF-1.5v was not able to activate caspases. Moreover, SF-1.5v induced glycogen synthase kinase (GSK) activation, but this protein is not involved in the heterofucan SF-1.5v induced apoptosis mechanism. In addition, ERK, p38, p53, pAKT and NFκB were not affected by the presence of SF-1.5v. We determined that SF-1.5v induces apoptosis in HeLa mainly by mitochondrial release of apoptosis-inducing factor (AIF) into cytosol. In addition, SF-1.5v decreases the expression of anti-apoptotic protein Bcl-2 and increased expression of apoptogenic protein Bax. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.     

Heterofucans from the brown seaweed Canistrocarpus cervicornis with anticoagulant and antioxidant activities

Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated l-fucose. We extracted six fucans from Canistrocarpus cervicornis by proteolytic digestion followed by sequential acetone precipitation. These heterofucans are composed mainly of fucose, glucuronic acid, galactose and sulfate. No polysaccharide was capable of prolonging prothrombin time (PT) at the concentration assayed. However, all polysaccharides prolonged activated partial thromboplastin time (aPTT). Four sulfated polysaccharides (CC-0.3/CC-0.5/CC-0.7/CC-1.0) doubled aPTT with only 0.1 mg/mL of plasma, only 1.25-fold less than Clexane, a commercial low molecular weight heparin. Heterofucans exhibited total antioxidant capacity, low hydroxyl radical scavenging activity, good superoxide radical scavenging efficiency (except CC-1.0), and excellent ferrous chelating ability (except CC-0.3). These results clearly indicate the beneficial effect of C. cervicornis polysaccharides as anticoagulants and antioxidants. Further purification steps and additional studies on structural features as well as in vivo experiments are needed to test the viability of their use as therapeutic agents.     

Genetic screening of FecB,FecXG and FecXI mutations and their linkage with litter size in Barki and Rahmani sheep breeds

Characterization of fecundity genes offers the opportunity to improve production efficiency, and the consequent increase in litter size in livestock industry, through utilizing them in breeding programs. The main objective of this study was to detect the BMPR‐IB, BMP15 and GDF9 gene mutations and to investigate whether these mutations are associated with litter size in Egyptian sheep breeds. To achieve this goal, 73 adult ewes representing Barki (n = 33) and Rahmani (n = 40) breeds were used. Polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) screening approach was used to detect the presence of FecB, FecX^G and FecX^I mutations in the two selected breeds. Results of this study showed that the three different candidate gene mutations, namely FecB, FecX^G and FecX^I are not present among these selected populations of the Egyptian breeds. Further studies regarding other mutations and/or other genes, which may influence ovulation rate, should be carried out to determine the type and mode of inheritance of such genes in Egyptian sheep breeds.     

Replacement of fish oil with vegetable oil blends in feeds for greater amberjack (Seriola dumerili) juveniles: Effect on growth performance,feed efficiency,tissue fatty acid composition and flesh nutritional value

This study was undertaken to assess the effects of fish oil (FO) substitution by a mixture of alternative vegetable oils (VO) on Seriola dumerili culture performance. A 154‐day feeding experiment was conducted using juveniles (39.2 ± 1.6 g average weight). Three isolipidic and isoenergetic meal‐based diets were formulated varying their lipid component. The control diet contained 100% FO (FO100), whereas diets VO50 and VO100 included 1/2 of oil blend and all the oil from blend of palm oil (PO) and linseed oil (LO) as substitute for FO, respectively. Dietary regime did not significantly affect growth performance, biometric indices, feed efficiency, plasma chemistry and liver and muscle lipid contents. Nonetheless, dietary VO inclusion impacted on the fatty acid profile of target tissues, especially in the liver. Fatty acid profiles of the fillets reflected those of the dietary oils except that there was apparent selective utilization of palmitic acid (C16:0) and oleic acid (C18:1n‐9) and apparent selective retention of long‐chain polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA, C20:5n‐3) and docosahexaenoic acid (DHA, C22:6n‐3). The nutritional value and the potential ability to prevent the development of coronary heart diseases of the flesh lipid fraction decreased with gradual FO substitution.     

Diversity of Brazilian biovar 2 strains of Ralstonia solanacearum

Ralstonia solanacearum is responsible for bacterial wilt disease. Specific and accurate identification of this pathogen is essential for protection of susceptible crops as well as breeding resistant varieties. Historically, R. solanacearum has been classified into biovars based on the use of sugar and alcohol as carbon sources, into races based on its ability to infect different hosts, more recently into phylotypes based on the intergenic transcribed sequence of the ribosomal RNA genes 16S and 23S and into sequevars based on the endoglucanase gene (egl) sequence. Race 3 biovar 2 (R3Bv2) is widespread in South and Central America, and in Brazil it is present in all potato-producing regions as the most prevalent strain. In this study, we classified 53 Brazilian R. solanacearum biovar 2 (Bv2) strains by traditional and molecular methods. PCR with specific primers confirmed all 53 bacterial strains as belonging to the R. solanacearum species complex, and all were classified as biovar 2A or 2T based on acidification of sugars and alcohols. Multiplex phylotype PCR assigned all strains to phylotype II. Phylogenetic analysis of egl sequences showed that most Bv2 strains from Brazil analyzed in this study did not cluster with known sequevars and are less clonal than the R3Bv2 strains reported for other countries. This is the first study to address the diversity of a collection of Brazilian R. solanacearum strains using the phylotype and sequevar classification scheme.     

Left-side displacement of the abomasum in dairy cows at pasture

The presentation of approximately 40 dairy cows affected with left-side displacement of the abomasum (LDA) per annum in a cattle practice in East Gippsland, Victoria provided an opportunity to conduct a survey and case-control study of the disease in a grazing environment. The study, involving 37 dairy cows at pasture, revealed significant differences from the pattern of the disease occurring in the northern hemisphere where cows in older age groups, of larger frame size, higher production and fed high grain rations are at increased risk. Affected cows were diagnosed over a 10-month period and represented approximately 0.06% of the dairy cow population. Most cases were diagnosed in the early lactation period. Evidence for a genetic predisposition was suggested by the discovery that one sire generated a disproportionately large number (9) of the cows with LDA. Although affected cows were average producers in their herds, being a member of a high-producing herd was a significant risk factor.