Impact of vitronectin concentration and surface properties on the stable propagation of human embryonic stem cells

The standard method for culturing human embryonic stem cells (hESC) uses supporting feeder layers of cells or an undefined substrate, Matrigel(?), which is a basement membrane extracted from murine sarcoma. For stem cell therapeutic applications, a superior alternative would be a defined, artificial surface that is based on immobilized human plasma vitronectin (VN), which is an adhesion-mediating protein. Therefore, VN adsorbed to diverse polymer surfaces was explored for the continuous propagation of hESC. Cells propagated on VN-coated tissue culture polystyrene (TCPS) are karyotypically normal after >10 passages of continuous culture, and are able to differentiate into embryoid bodies containing all three germ layers. Expansion rates and pluripotent marker expression verified that a minimal VN surface density threshold is required on TCPS. Further exploration of adsorbed VN was conducted on polymer substrates with different properties, ranging from hydrophilic to hydrophobic and including cationic and anionic polyelectrolyte coatings. Despite differing surface properties, these substrates adsorbed VN above the required surface density threshold and were capable of supporting hESC expansion for >10 passages. Correlating wettability of the VN-coated surfaces with the response of cultured hESC, higher cell expansion rates and OCT-4 expression levels were found for VN-coated TCPS, which exhibits a water contact angle close to 65°. Importantly, this simple, defined surface matches the performance of the benchmark Matrigel, which is a hydrogel with highly complex composition.     

Diapause in Scelio chortoicetes Frogg

Ohne Zusammenfassung     

Survival and fertility after uterine prolapse in dairy cows

Sixty-eight cases of uterine prolapase in pastured dairy cows were treated in 2 consecutive spring calving seasons in East Gippsland, Victoria. Fifty cows survived (73.5%). Of 43 cows available for followup, 36 (84%) conceived in the mating period following the prolapse, taking 10 d longer to conceive than herd mates that calved on the same day. Three of the 36 cows (8%) that conceived, aborted, this occurring in the middle trimester of pregnancy. No prolapses occurred at the following calving but one case had suffered uterine prolapse 2 years previously. The conclusions drawn from these observations are that cows with uterine prolapse have a good chance of surviving if treated, that treatment is cost-effective, that uterine prolapse is unlikely to reoccur and treated cows have a good chance of conceiving. The veterinarians involved in this investigation were reasonably accurate in their ability to predict long term survival but not as good in predicting ability to conceive again.     

Vaccine efficacy for reducing turbinate atrophy and improving growth rate in piggeries with endemic atrophic rhinitis

Two vaccines, based on formalin-killed whole cells of toxigenic Pasteurella multocida type D and Bordetella bronchiseptica combined with a partially toxoided cell extract of P multocida, were prepared with Freund's incomplete adjuvant (vaccine 1) or by alum precipitation (vaccine 2). Each was tested for safety and efficacy in reducing the severity of nasal turbinate atrophy and improving the growth rate of pigs in three Western Australian commercial piggeries with endemic atrophic rhinitis. In safety experiments with vaccine 1, no adverse clinical effects were observed in vaccinated sows or their progeny. Piglets receiving vaccine 2 showed no injection site abnormalities, pyrexia or turbinate atrophy. In field trials, vaccine 1 significantly reduced the prevalence of moderate to severe nasal turbinate atrophy (Done score 3 to 5) when used in two piggeries (A and B). Progeny from vaccinated sows in piggery B also grew significantly faster than controls. When vaccine 2 was used in piggery A at a later date and in another piggery (C), growth rate was not improved in either piggery and the prevalence of moderate to severe turbinate atrophy was reduced only in piggery C.     

Supplementation with sunflower seeds in beef cattle did not impact on oocyte and in vitro embryo production

Supplementation with compounds rich in linoleic acid, including sunflower seed supplementation, promotes increase in conception rates in cows. We aimed to evaluate whether the sunflower seed (linoleic acid source) supplementation in beef donor females alters the plasma concentrations of cholesterol, triglycerides, HDL and LDL, increases the number and quality of oocytes, increases the cleavage rates and determines an improvement in number and quality of in vitro produced blastocysts. Thus, Nelore females were divided into two groups of 15 animals to receive supplementation with or without sunflower seed for 57 days. Females underwent follicular aspiration and the oocytes were subjected to in vitro embryo production. There was no difference (p > .1) between control group and group supplemented with sunflower seed on the number of displayed follicles; number of aspired oocytes; recovery rate; cleavage rate; number of embryos; number of blastocysts; embryos number of grades I and II; plasma concentrations of total cholesterol, triglycerides; HDL and LDL. Therefore, sunflower seed supplementation in oocyte donors did not increase the number and quality of oocytes, cleavage rates and the number and quality of blastocysts produced in vitro.     

Effects of the supplementation with an high‐polyphenols extra‐virgin olive oil on kinetic sperm features and seminal plasma oxidative status in healthy dogs

The aim of this work was to evaluate the effects of the supplementation of two extra‐virgin olive oils (EVOO) having different polyphenols content, on canine spermatozoa kinetic parameters and seminal plasma oxidative status. The study was conducted on 12 clinically healthy dogs of different breeds (2–7 years, 5–48 kg of body weight) divided into two groups: an experimental group supplemented with EVOO (Coratina cultivar) high in polyphenols (H‐P) and a control group fed EVOO (Cima di Bitonto cultivar) low in polyphenols (L‐P). The oil was daily administered per os (1 ml/3 kg BW) before meal. Semen collection was made twice at 15 days distance (D0₁ and D0₂) and then at 30 (D30), 60 (D60) and 90 (D90) days. Semen concentration and kinetic parameters were measured using computer‐assisted sperm analysis (CASA) system to evaluate: sperm total count, sperm motile (MOT%), progressive motility (PROGR%) and its fractions, straight‐line velocity (VSL, μm/s), curvilinear velocity (VCL, μm/s), average path velocity (VAP, μm/s), amplitude of lateral head displacement (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR%) and linearity (LIN%). On seminal plasma, reactive oxygen species (ROS) and biological antioxidant potential (BAP) were tested. From findings, no differences were found for sperm MOT, VSL, VCL, VAP, ALH, BCF, STR, LIN and BAP. A gradual enhancement of PROGR% was observed in H‐P group (p < .01). The ROS levels were higher in dogs H‐P compared to the other group (p < .05). In conclusion, our results highlight the positive effects of EVOO polyphenols on sperm PROGR% in healthy dogs.     

Chemical composition and the nutritive quality of different wheat cultivars for broiler chickens

1. Different wheat cultivar samples were grown in replicated trials on a single site in three harvest years. A total of 23 wheat samples were harvested. Nutritionally complete, meal-form diets that included each wheat sample at 650 g/kg were used to compare broiler growth performance and determine apparent metabolisable energy (AME). The relationships were examined between these variables of nutritive value for broilers and the chemical composition and tests of quality on the wheat samples. 2. The total starch contents of the wheat samples ranged from 594 to 732 g/kg dry matter (DM). The mean total non-starch polysaccharide (NSP) content was 100g/kg DM comprising 73.5 g/kg DM of insoluble material. Endosperm hardness ranged between 10 and 77 relative units and Beaver was the softest cultivar in two of the batches of samples. The AME of the wheat-based diets had a maximum range of 0.5 MJ/kg within each of the three batches of wheat that were tested. 3. Endosperm hardness of the wheat grains, within a harvest year, was positively correlated with broiler weight gain and the 1000-grain weight was negatively correlated with feed conversion efficiency (FCE). Starch content and gross energy were positively correlated with determined AME, and DM, 1000-grain weight and water holding capacity were negatively correlated with AME. 4. A step-wise regression technique indicated that endosperm hardness and ash content of the wheat were the explanatory variables that, within harvest years, significantly reduced the unexplained variation in broiler growth rate, feed intake and FCE. The contents of total starch, crude protein and ether extract were the explanatory variables that, within harvest years, significantly reduced the unexplained variation in AME.