Proteomic analysis of amniotic and allantoic fluid from buffaloes during foetal development

The objective of this study was to describe the dynamic changes in protein composition and protein abundance in amniotic and allantoic fluids from buffaloes during gestation. Amniotic and allantoic fluids were collected during the first, second and third trimesters of gestation. The foetuses were measured and weighed. Fluid samples were centrifuged at 800 g for 10 min and then at 10,000 g for 60 min at 4°C. The supernatant was collected to determine the total protein concentration. Based on total protein concentration, an aliquot (50 μg) was used for in‐solution tryptic digestion, and mass spectrometry analysis (nano‐LC‐MS/MS) was performed. A multivariate statistical analysis of the proteomic data was conducted. Across the different stages of buffalo gestation, fifty‐one proteins were found in the amniotic fluid, and twenty‐one were found in the allantoic fluid. A total of twelve proteins were common among the stages, and four presented significant differences (VIP score α > 1). Fibronectin and alpha‐1‐antiproteinase were more abundant in the amniotic fluid than in the allantoic fluid. Alpha‐2‐macroglobulin and alpha‐2‐HS‐glycoprotein were more abundant in the allantoic fluid than in the amniotic fluid. Alpha‐2‐macroglobulin participates in remodelling and growth of the uterus at beginning of the gestation (first trimester), and these findings indicate that can serve as a potential tool for the early diagnosis of pregnancy in buffaloes.     

Differential interaction between two Glomus intraradices strains and a phosphate solubilizing bacterium in maize rhizosphere

Arbuscular mycorrhizal (AM) fungi and phosphate solubilizing bacteria (PSB) have a positive effect on plant productivity primarily through increasing phosphate availability. In order to study the interaction between AM fungi and PSB, we used Bacillus megaterium, a PSB isolated from the sterilized surface of AM germinated spores, and two strains of the AM fungus Glomus intraradices with different mycelial architecture. A greenhouse experiment was designed with maize as host plant with the addition of tribasic calcium phosphate. We tested the hypothesis that PSB, intimately linked with AM fungi, could interact differentially with the two AM strains. We concluded that inoculation with the PSB positively affected maize mycorrhization. Insoluble phosphate alone did not influence the AM extraradical mycelium (ERM) length and maize mycorrhization when bacteria were not inoculated. The results provide evidence that the adverse effect on infectivity for some AM strains might be caused by solubilized phosphorus release to the rhizosphere by PSB. Differences related to the mycelium architecture of each AM strain were observed: the density of PSB in rhizosphere soil was significantly higher only with the GA8 strain coinciding with the highest values of maize biomass. The density of bacteria associated with GA8 mycelium could be the result of the transfer of photosynthates through the rhizosphere; this close contact would favor the persistence of the intimate relationship between PSB and AM hyphae. In the bacteria-free treatments, soil adherence was not significantly altered. Although the highest development of ERM occurred with GA5, plants inoculated with GA8 showed the highest values for soil adherence. This may be due to the AM mycelium which modifies bacterial persistence in the rhizosphere and consequently soil adherence. Our results show that for potential applications, some characteristics of the AM strains are key in the selection of the AM fungi–PSB combinations. These include the tolerance to soluble phosphorus, the rate of root colonization, and ERM development that favors the persistence of bacteria in rhizosphere soil.     

Chemical interesterification of blends of palm stearin, coconut oil, and canola oil: physicochemical properties

trans-Free interesterified fat was produced for possible usage as a margarine. Palm stearin, coconut oil, and canola oil were used as substrates for chemical interesterification. The main aim of the present study was to evaluate the physicochemical properties of blends of palm stearin, coconut oil, and canola oil submitted to chemical interesterification using sodium methoxide as the catalyst. The original and interesterified blends were examined for fatty acid composition, softening and melting points, solid fat content, and consistency. Chemical interesterification reduced softening and melting points, consistency, and solid fat content. The interesterified fats showed desirable physicochemical properties for possible use as a margarine. Therefore, our result suggested that the interesterified fat without trans-fatty acids could be used as an alternative to partially hydrogenated fat.     

Performance of Nellore heifers, forage mass, and structural and nutritional characteristics of Brachiaria brizantha grass in integrated production systems

Our objective was to evaluate production, nutritive value and carrying capacity of piatã grass (Brachiaria brizantha cv. BRS Piatã), and performance of Nellore heifers in agrosilvopastoral systems (ASPS) with three eucalyptus (Eucalyptus urograndis) tree densities, during winter, spring, summer, and fall. Three integrated systems were evaluated: ASPS-1 (357 trees ha^?1), ASPS-2 (227 trees ha^?1), and CON (5 trees ha^?1). In each system, two sward heights were evaluated: short and tall. A total of 80 11-month-old Nellore heifers were randomly allocated in a randomized split-plot block, 3?×?2 factorial. Greater dry matter availability was observed on CON pastures during the fall season. Greater percentage of leaf lamina was detected on ASPS-1 with short sward height and greater during summer, compared with other seasons. A greater forage production was observed between tree rows and for tall sward height. Spring was the season with less forage nutritive value. Average daily gain was greater during summer and fall. Gain per hectare and stocking rate were greater on CON system and on ASPS-2. Pastures with short sward height had greater gain per hectare and stocking rate. Agrosilvopastoral systems with intermediate tree density seem to be a good choice for producers willing to diversify their revenue sources without decreasing animal production.     

Mutations that increase the life span of C. elegans inhibit tumor growth

Mutations in gld-1 cause lethal germline tumors in the nematode Caenorhabditis elegans. We find that a wide variety of mutations that extend C. elegans' life span confer resistance to these tumors. The long life spans of daf-2/insulin-receptor mutants were not shortened at all by gld-1 mutations; we attribute this finding to decreased cell division and increased DAF-16/p53-dependent apoptosis within the tumors. Mutations that increase life span by restricting food intake or inhibiting respiration did not affect apoptosis but reduced tumor cell division. Unexpectedly, none of these longevity mutations affected mitosis in normal germlines; this finding suggests that cellular changes that lead to longevity preferentially antagonize tumor cell growth.     

In vitro activity of VEGF-E produced by orf virus strains isolated from classical and severe persistent contagious ecthyma

Proliferative orf virus infections in adult sheep have increased in Italy in the past few years: these extreme cases are frequently fatal and difficult to differentiate from other infectious diseases of sheep such as blue tongue. A probable explanation for the proliferative and highly vascularized nature of the lesions was found in the expression of the VEGF-E gene encoded by the orf virus. To investigate a possible role of the viral VEGF in the pathogenesis of severe persistent orf virus lesions, the activity of four VEGF-E variants was compared by an angiogenesis in vitro model. Similar angiogenic activity was found between strains isolated from the classical and the proliferative forms of the disease, even if the latter was able to develop a higher number of vessels during the first 24 h of infection. Our in vitro findings seems to exclude that the VEGF variants encoded by the strain isolated from the atypical form of the disease could be the responsible for the histopathological aspect of the proliferative lesions.     

Occurrence of ochratoxin A- and aflatoxin B1-producing fungi in Lebanese grapes and ochratoxin a content in musts and finished wines during 2004

This paper reports the results of an extensive survey on the occurrence of filamentous fungi isolated from wine-grapes in Lebanon and to test their ability to produce ochratoxin A (OTA) and aflatoxin B1 (AFB1) on CYA culture medium, in order to assess their potential for producing these mycotoxins on grapes. From the 470 grapes samples taken during season 2004, 550 fungi strains were isolated with 490 belonging to Aspergillus spp. and 60 belonging to Penicillium spp. All these isolated fungi starins were tested for their ability to produce OTA and AFB1. Aspergillus carbonarius shows that it is the only species able to produce OTA with a production percentage reaching 100% and a maximum concentration of 52.8 microg/g of Czapek yeast extract agar (CYA). In its turn, Aspergillus flavus was considered as the only AFB1-producing species with production percentage of 45.3% and a maximum concentration reaching 40 microg/g CYA. A total of 47 handmade musts produced from the collected grapes were also analyzed in order to correlate the presence of OTA in must and the occurrence of filamentous fungi on grapes; 57.4% were contaminated with OTA at low level with concentrations ranging between 0.011 and 0.221 microg OTA L(-1). The analysis of these must samples was not performed with regard to AFB1. Seventy samples of finish red wine were also assayed for OTA content. The results showed that 42 of the tested samples (60%) were found to be positive for OTA with low levels (0.012-0.126 microg OTA L(-1)).     

Carotenoid composition of hydroponic leafy vegetables

Because hydroponic production of vegetables is becoming more common, the carotenoid composition of hydroponic leafy vegetables commercialized in Campinas, Brazil, was determined. All samples were collected and analyzed in winter. Lactucaxanthin was quantified for the first time and was found to have concentrations similar to that of neoxanthin in the four types of lettuce analyzed. Lutein predominated in cress, chicory, and roquette (75.4 +/- 10.2, 57.0 +/- 10.3, and 52.2 +/- 12.6 microg/g, respectively). In the lactucaxanthin-containing lettuces, beta-carotene and lutein were the principal carotenoids (ranging from 9.9 +/- 1.5 to 24.6 +/- 3.1 microg/g and from 10.2 +/- 1.0 to 22.9 +/- 2.6 microg/g, respectively). Comparison of hydroponic and field-produced curly lettuce, taken from neighboring farms, showed that the hydroponic lettuce had significantly lower lutein, beta-carotene, violaxanthin, and neoxanthin contents than the conventionally produced lettuce. Because the hydroponic farm had a polyethylene covering, less exposure to sunlight and lower temperatures may have decreased carotenogenesis.     

First successful frozen semen of the maned wolf (Chrysocyon brachyurus)

This study aimed to describe successful cryopreservation of sperm from maned wolves (Chrysocyon brachyurus). Three ejaculates from 2 maned wolves were collected by digital manipulation of the penis and evaluated subjectively, centrifuged and frozen in BotuCrio^® (Botupharma, Botucatu, Brazil) or Tris–yolk egg extender. Spermatozoa were thawed at 37ºC/30s or 70ºC/4s and evaluated for kinetics, morphology, plasma and acrosome membrane integrity, mitochondrial potential, hydrogen peroxide, superoxide anion and lipid peroxidation. From 5 thawed samples, two had sperm total motility >55% (56.0% and 64.0%) and progressive motility ~35% (35% and 40%), both frozen with Tris–yolk egg. Plasma and acrosome membrane integrity decreased and percentage of sperm defects increased post-thawing. We concluded that is possible to freeze spermatozoa from maned wolves using semen collection and processing methods applied for domestic dogs.     

Vitrification of canine ovarian tissue using the Ovarian Tissue Cryosystem (OTC) device

The present study evaluated the effect of Ovarian Tissue Cryosystem (OTC) on follicular morphology and density, as well as on stromal cell density of vitrified canine ovarian tissue. Canine ovarian fragments collected from adult female dogs in stages of the random oestrous cycle were fixed (FC, fresh control) or vitrified (VIT) with an OTC device. After vitrification and warming, the fragments were fixed for histological analysis. Overall, the mean percentage of normal pre-antral follicles decreased after vitrification procedure (FC: 74.5% ± 1.6% vs. VIT: 52.05% ± 1.5%). Although the rates of normal primordial (71.1% ± 1.8%) and secondary (0.7% ± 0.4%) follicles vitrified showed a reduction (p < .05), vitrification using OTC showed considerable preservation of follicles, when compared to the fresh control (81.1% ± 1.5% and 2.3% ± 0.6%, respectively). The mean follicular density was maintained after vitrification (FC: 199.65 ± 12.8 vs. VIT: 199.68 ± 10.8), whereas the stromal cell density decreased in the VIT group. Based on the results, we recommend the use of OTC for vitrification of canine ovarian tissue.