Dummy regression analysis for modelling the nutritionally tailored fillet fatty acid composition of turbot and sole using gilthead sea bream as a reference subgroup category

Farmed turbot and sole were sampled at different stages of the production cycle for analysis of fillet lipid content and fatty acid (FA) composition. The entire data set along with our own published data on gilthead sea bream were fitted to dummy regression equations with turbot and sole as dummy variables, gilthead sea bream as a reference subgroup category, and diet FA composition and fillet lipid content as independent variables. The relative contribution of each independent variable to the total variance was found to vary within and among FAs and fish species, but strong correlation coefficients (0.76 <  r² > 0.99) were found for almost all of the FA equations, including saturated FAs, monoenes and long‐chain polyunsaturated fatty acids (PUFA) of n‐3 and n‐6 series. Given the differences in lipogenic activities of the fish species, major interaction effects between fillet lipid content and dummy variables were found for monoenes and saturated FAs. The proposed equations (hosted at www.nutrigroup-iats.org/aquafat ) were able to fit different proportions of EPA, DPA and DHA underlying the fish species differences in FA desaturation/elongation pathways. The robustness of the model was proven with extra data from the three fish species, allowing a close linear association near to equality for the scatter plot of observed and predicted values.     

Isolation of Canine Mammary Cells With Stem Cell Properties and Tumour-Initiating Potential

Recent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self-renewal or by progenitor cells that have acquired the capacity for cell self-renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long-term non-adherent culture conditions. We showed that cells obtained from spheres that display self-renewing properties, have multi-lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour-initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.     

Equine amnionitis and fetal loss: The case definition for an unrecognised cause of abortion in mares

A series of abortions occurred in mares in New South Wales during 2004 that involved similar and unusual findings on post mortem examination of aborted fetuses and fetal membranes. The term Equine Amnionitis and Fetal Loss (EAFL) was developed to describe the condition. This form of abortion had not been previously recognised in Australia. The pathology alone is not specific for EAFL and diagnosis requires demonstration of a combination of certain pathological and bacteriological features. The purpose of this paper is to describe patterns considered consistent with EAFL cases as a working case definition for use by veterinarians and veterinary pathologists in identifying future cases of EAFL. More detailed papers are in preparation to fully describe the epidemiological, histopathological, and microbiological aspects of EAFL.     

The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa

To try new acrosomal probes for assessing ram spermatozoa, we compared the LysoSensor^? probe, which labels acidic organelles, with the frequently used peanut agglutinin acrosomal probe (PNA‐PE; phycoerythrin as fluorescent moiety). The previous microscopic observations showed a lack of relationship of LysoSensor^? with acrosomal status. Semen was obtained from five rams and frozen in four pools. Each pool was analysed carrying out a triple staining propidium ioide/PNA‐PE/LysoSensor^? Green DND‐189 to test acrosome labelling, and a double staining SYBR‐14/PI, to assess sperm viability. Stained samples were analysed by flow cytometry. All measurements were replicated. Data were processed using agreement and repeatability tests. LysoSensor^? labelling did not agree with PNA (mean of differences: 30.8%; coefficient of agreement: 22.6%), confirming microscopic observations. Nevertheless, when LysoSensor^? was compared with SYBR‐14/PI, the agreement was high (mean of differences: ?0.05%; coefficient of agreement: 5.07%). Repeatability of both methods was high and similar. LysoSensor^? did not seem to specifically stain the acrosome, but it may accumulate in the cytoplasm and label viable spermatozoa. Therefore, LysoSensor^? might not be used as an acrosomal probe in ram spermatozoa, but it could be used in other kind of studies, taking advantage of its pH sensitivity.     

Improvement of Fertility in Artificially Inseminated Ewes Following Vaginal Treatment with Misoprostol Plus Terbutaline Sulphate

The effect of vaginal administration of misoprostol plus terbutaline sulphate 6 h prior to artificial insemination (AI) upon the site of AI (vaginal or cervical) and fertility was studied using a total of 87 estrous synchronized Serra da Estrela ewes (control n = 42 and treated n = 45). Artificial insemination was performed using refrigerated semen at 54–55 h after sponge removal. Lambing rate (fertility) and prolificacy were compared between control and treated ewes. The effect of the site of semen deposition on fertility was also evaluated. Prolificacy rate was not different between control (1.5) and treated (1.59) ewes. The proportion of cervical AI achieved in control (45.2%) and treated (37.8%) ewes was not significantly different. Overall, fertility was significantly lower in control than in treated ewes (42.9% vs 64.4%; p < 0.04). Fertility following vaginal AI was significantly lower for control for than treated ewes (30.4% vs 60.7%; p < 0.03) but the difference was smaller and not significant for cervical AI (control 57.9% vs 70.6%). It was concluded that vaginal administration of misoprostol plus terbutaline sulphate 6 h prior to artificial insemination did not affect the proportion of cervical inseminations but significantly improved the fertility of treated ewes. Although needing confirmation, it was hypothesized that drugs might have induced local secretory modifications leading to an increase of cervical ability to retain more viable spermatozoa for fertilization.     

Effect of quercetin on platelet spreading on collagen and fibrinogen and on multiple platelet kinases

α₂β₁ and α_IIbβ₃ integrins, that support platelet adhesion to collagen and fibrinogen, respectively, share common signaling molecules. The effect of quercetin on platelet static adhesion to collagen and fibrinogen was assessed and correlated with its kinase inhibitory activity. Quercetin strongly abrogated PI3K and Src kinases, mildly inhibited Akt1/2, and slightly affected PKC, p38 and ERK1/2. Quercetin or the combined use of adenosine diphosphate and thromboxane A₂ inhibitors abrogated platelet spreading on these surfaces to a similar extent. We suggest that the inhibitory effect of quercetin on platelet kinases blocks early signaling events preventing a complete platelet spreading.     

VARIATION IN SOIL ORGANIC CARBON AND NITROGEN STOCKS ALONG A TOPOSEQUENCE IN A TRADITIONAL MEDITERRANEAN OLIVE GROVE

The impact of the topographical position on soil properties was evaluated in an olive grove with traditional tillage. Three topographical positions: summit, backslope and toeslope were chosen for evaluation. The soil samples were taken from four soil sections of 0·25 m (0–1 m). The soil organic carbon (SOC) and N content increased along the downslope direction (5·5, 6·5 and 7·1 g C kg⁻¹ and 0·3, 0·8 and 0·9 g N kg⁻¹ in the surface layer in the summit, backslope and toeslope respectively) as well as SOC and N stocks, considering the two first soil sections. In addition, there was movement of the most erodible textural fraction (silt). However, the total SOC stock (refer to 1 m of depth) did not vary with respect to the topographical position, but the total N stock (refer to 1 m of depth) varied significantly. These increases were due to erosion processes that occur along the toposequence, leading to organic matter transfers from the summit to the toeslope. All the stratification ratios calculated were lower than 2, indicating the low quality of the soils. Therefore, alternative management techniques that avoid soil erosion must be considered in olive grove in order to increase the soil quality and fertility. Copyright © 2014 John Wiley & Sons, Ltd.     

Selected Metabolic and Hormonal Profiles during Maintenance of Spontaneous Ovarian Cysts in Dairy Cows

Information is lacking regarding the relationship between metabolic and hormonal profiles and the maintenance of spontaneous ovarian cyst disease in dairy cows. For this reason, the concentrations of non‐esterified fatty acids (NEFA), insulin‐like growth factor I (IGF‐I) and cortisol (C) were investigated during the spontaneous course of ovarian cyst disease in dairy cows (n = 6) between the 7th and 16th weeks post‐partum (PP). The control group consisted of normally cycling cows (n = 6). Blood samples were collected twice a day, and plasma was analysed using different techniques. Progesterone and 15‐ketodihydro‐PGF_2α plasma profiles were investigated to confirm the ovulatory or anovulatory conditions of the cows. Cortisol plasma levels were not significantly different among sampling times within each group or between the two groups. NEFA plasma levels were significantly higher in cycling cows compared to cystic cows at the 16th week PP (p < 0.01), but with rather low values, indicating by now sparse mobilization of fat stores. Insulin‐like growth factor I plasma concentrations were higher in cystic cows during the 8th, 10th, 11th (p < 0.01) and 16th week PP (p < 0.05), indicating that the presence of ovarian cysts coincides with increased IGF‐I levels. These results suggest no influence of cortisol and NEFA levels in cysts maintenance, while a possible involvement of IGF‐I can be suspected not only in the pathogenesis, as already known, but also in the maintenance of spontaneous cystic ovarian disease in cattle.