Expression of TGFBR1, TGFBR2, TGFBR3, ACVR1B and ACVR2B is altered in ovaries of cows with cystic ovarian disease

The objective of this study was to examine the expression of transforming growth factor beta receptor (TGFBR)1, TGFBR2, TGFBR3, activin receptor (ACVR)1B and ACVR2B in ovaries of cows with cystic ovarian disease (COD). The expression of the selected receptors was determined by immunohistochemistry in sections of ovaries from cows with ACTH‐induced and spontaneous COD. Expression of TGFBR1 and TGFBR3 was higher in granulosa cells of cysts from cows with spontaneous COD than in tertiary follicles from the control group. Additionally, TGFBR3 expression was higher in granulosa cells of cysts from cows with ACTH‐induced COD than in those from the control group and lower in theca cells of spontaneous and ACTH‐induced cysts than in tertiary control follicles. There were no changes in the expression of TGFBR2. ACVR1B expression was higher in granulosa cells of tertiary follicles of cows with spontaneous COD than in the control group, whereas ACVR2B expression was higher in cysts of the spontaneous COD group than in tertiary follicles from the control group. The alterations here detected, together with the altered expression of the ligands previously reported, indicate alterations in the response of the ligands in the target cells, modifying their actions at cellular level.     

Sperm Susceptibility to Oxidative Stress in the Retuertas Endangered Horse

In order to determine whether the high level of inbreeding of the Retuertas endangered Horses may increase the vulnerability to oxidative stress of the sperm, the susceptibility to lipid peroxidation (LPO) of sperm membranes was evaluated in cooled extended ejaculates. Extended ejaculates (INRA 96) from four Retuertas horses and nine ejaculates from pure Spanish horses were received within 24 hours of collection at our laboratory. An aliquot was analyzed and served as time 0 control. The rest of the ejaculates were split into two additional aliquots: the first aliquot served as an incubation control and the second aliquot received an oxidative insult (Fe₂SO₄). Membrane LPO, motility parameters, membrane integrity, and mitochondrial membrane potential were evaluated. The oxidative insult induced a significant increase in the percentage of sperm with LPO in the Retuertas horses (P < .05). Total and progressive motility were not affected by the oxidative insult in both groups. However, curvilinear velocity (VCL), average velocity (VAP), straightness index (STR), and amplitude of lateral head displacement (ALH) were significantly affected by the treatment (P < .05) in the Retuertas horse population. The membrane integrity of sperm in this group was also affected: the percentage of sperm with intact membranes decreased (57.31% ± 7.76% vs. 47.52% ± 8.28%); and the percentage of necrotic sperm increased (33.27% ± 7.83% vs. 44.85% ± 9.16%) (P < .05). Finally, the oxidative insult increased the population with low mitochondrial membrane potential in these wild horses. We concluded that the high level of inbreeding of this particular breed apparently leads to a higher susceptibility to lipid peroxidation.     

The use of gelatine in long‐term storage (up to 48 hr) at 5°C preserves the pre‐freezing and post‐thawing quality of brown bear sperm

Sedimentation of spermatozoa occurs during long‐term liquid storage and this may produce deleterious changes. Our aim was to apply gelatine supplementation during long‐term pre‐freezing storage of bear sperm, applying final dilution and 6% glycerol at room temperature and cool in straws. We tested four models of sperm storage using a 1:1 dilution in TTF‐ULE‐Bear extender (TesT‐fructose‐egg yolk‐glycerol 6%): (i) second 1:1 dilution at room temperature (RT), cooling at 5°C in a tube and final dilution (100 × 10⁶ sperm ml^?1) (Standard); (ii) final dilution at RT and cooling in a tube (FD‐Tube); (iii) final dilution at RT and cooling in 0.25 ml plastic straw (FD‐Straw); and (iv) final dilution at RT in extender supplemented with 1.5% gelatine (Gelatine) and cooling in a 0.25 ml plastic straw. A Standard sample was stored at 5°C for 1 hr (Control); the rest of the samples (Standard, FD‐Tube, FD‐Straw, Gelatine) were stored for 24 or 48 hrs before freezing (100 × 10⁶ sperm ml^?1, glycerol 6%). The quality of the samples was assessed for motility by CASA, and viability (SYBR‐14/propidium iodide‐PI‐; VIAB), acrosomal status (PNA‐FITC/PI; iACR) and apoptotic status (YO‐PRO‐1/PI; YOPRO‐) by flow cytometry. At pre‐freezing, after 48 hr, Gelatine showed significantly higher viability (for VIAB and YOPRO‐) and progressiveness (PM, LIN and STR). At 48 hr, Gelatine showed similar YOPRO‐, iACR, LIN, STR and ALH respect to Control. At both 24 and 48 h post‐thawing, Gelatine sample had similar scores for YOPRO‐, iACR, LIN, STR, WOB and VIAB (only 24 hr) when compared with Control, and lower for TM, PM, rapidPM, VAP and ALH. No differences were found among others experimental groups with respect to Control. In conclusion, gelatine could be a suitable alternative to preserve the viability and progressive motility of brown bear ejaculates during long‐term pre‐freezing storage at 5°C.     

Distribution of Infectious Pancreatic Necrosis Virus (IPNV) Based on Surveillance Programs in Freshwater Trout Farms of Mexico

Diagnostic testing was performed between 2000 and 2012 to determine the distribution of infectious pancreatic necrosis virus (IPNV) in the main states of the Mexican Republic with freshwater Rainbow Trout Oncorhynchus mykiss (Walbaum) farms. This virus was positively identified from Rainbow Trout farms in seven of the eight states assessed. Due to nonnormal data distribution, a logistic regression model was applied for statistical analysis, the results of which indicated that virus prevalence was variable between states, with moderate but significant differences. Regarding the time periods evaluated, IPNV prevalence was higher during the first years of the study. The susceptible, infected, removed model was used to examine this phenomenon, which indicated that the decreased prevalence during the latter years of the study could be associated with a real elimination of the infection. The information of the cases analyzed also suggests a relationship with the irregularity in the submission of samples to the laboratory and emphasizes other factors that have contributed to the transmission of IPNV throughout the country.

Received November 10, 2014; accepted December 5, 2015.     

Effectiveness of mycorrhizal inoculation in the nursery on growth and water relations of Pinus radiata in different water regimes

We compared the performance of mycorrhiza-inoculated and non-inoculated radiata pine (Pinus radiata D. Don) seedlings in two field studies. In the first study, above ground growth of 2-year-old container-grown trees was compared after planting in two areas that differed in water availability. Mycorrhizal inoculation significantly improved above ground growth in the 2 years after planting in both areas, especially at the drier site. In the second study, the influence of Rhizopogon roseolus (Corda ex Sturm) Th. Fries and Scleroderma citrinum Pers. on xylem water potential and hydraulic conductance at the soil-root interface of trees established at the same sites was assessed. Nursery inoculations with R. roseolus and S. citrinum improved tree growth during the first 2 years after field outplanting, particularly at the drier site.     

The Impact of Reproductive Technologies on Stallion Mitochondrial Function

The traditional assessment of stallion sperm comprises evaluation of sperm motility and membrane integrity and identification of abnormal morphology of the spermatozoa. More recently, the progressive introduction of flow cytometry is increasing the number of tests available. However, compared with other sperm structures and functions, the evaluation of mitochondria has received less attention in stallion andrology. Recent research indicates that sperm mitochondria are key structures in sperm function suffering major changes during biotechnological procedures such as cryopreservation. In this paper, mitochondrial structure and function will be reviewed in the stallion, when possible specific stallion studies will be discussed, and general findings on mammalian mitochondrial function will be argued when relevant. Especial emphasis will be put on their role as source of reactive oxygen species and in their role regulating sperm lifespan, a possible target to investigate with the aim to improve the quality of frozen–thawed stallion sperm. Later on, the impact of current sperm technologies, principally cryopreservation, on mitochondrial function will be discussed pointing out novel areas of research interest with high potential to improve current sperm technologies.     

First Web-Based Database on Total Phenolics and Oxygen Radical Absorbance Capacity (ORAC) of Fruits Produced and Consumed within the South Andes Region of South America

This paper reports the first database on antioxidants contained in fruits produced and consumed within the south Andes region of South America. The database ( www.portalantioxidantes.com ) contains over 500 total phenolics (TP) and ORAC values for more than 120 species/varieties of fruits. All analyses were conducted by a single ISO/IEC 17025-certified laboratory. The characterization comprised native berries such as maqui ( Aristotelia chilensis ), murtilla ( Ugni molinae ), and calafate ( Barberis microphylla ), which largely outscored all other studied fruits. Major differences in TP and ORAC were observed as a function of the fruit variety in berries, avocado, cherries, and apples. In fruits such as pears, apples, apricots, and peaches, a significant part of the TP and ORAC was accounted for by the antioxidants present in the peel. These data should be useful to estimate the fruit-based intake of TP and, through the ORAC data, their antioxidant-related contribution to the diet of south Andes populations.     

Effect of β-endorphin on adherence, chemotaxis and phagocytosis of Candida albicans by peritoneal macrophages

There is growing evidence about the role of neuroendocrine hormones in the regulation of the immune system. In the present study we have examined effects on different stages of phagocytic function of peritoneal macrophages from BALB/c mice induced by β-endorphin. Peritoneal macrophages were incubated (30 min at 37°C) in vitro with 0.22, 0.5 or 2200 ng/ml of this hormone. Adherence capacity was evaluated by means of a substrate adherence technique, chemotaxis in Boyden chambers, and ingestion of Candida albicans on migration inhibitory factor (MIF) dishes. No changes in adherence capacity were found. Chemotaxis, however, increased, and concentration of β-endorphin correlated directly with stimulation. Incubation of macrophages with 0.5 ng/ml of β-endorphin also stimulated phagocytosis of Candida albicans. These results indicate that β-endorphin acts on peritoneal murine macrophages, stimulating some stages of their phagocytic function.     

Management of water consumption in agriculture A model for the economic optimisation of water use: application to a sub-humid area

This paper present a set of solutions for new irrigation transformation in a sub-humid area such as Salvatierra (Alava, Spain). The research is based on the choice of crop rotation (cultivated species and its degree of participation) being able to economically optimise the use of available water for irrigation. Hence, we will be able in helping make decisions to plan, from origin, the transformation of a dry area into an irrigated area. To do this, a model of economic use of water has been used in an area with climatic features very similar to a large part of Europe, representing an interesting scenario compared to the places where the model had not been applied. Crops taking part in this rotation to optimise use of water are mainly vegetables, with high water needs, that coexist together with crops for dryland farming even in conditions of lack of water. Crops for dryland farming allow an interesting diversification of activity as well as an average resource assignment to the farms that make possible to cultivate many farms at a time, which obviously implies socio-economic benefits for the achievements in the zone.     

Rapid detection of Fusarium oxysporum f. sp. lactucae on soil,lettuce seeds and plants using loop‐mediated isothermal amplification

Fusarium oxysporum f. sp. lactucae (FOL) is a soil‐ and seedborne pathogen and the causal agent of fusarium wilt on lettuce. Four races have been identified within FOL, with different worldwide distribution. Several molecular techniques have been used to detect and identify this pathogen; however, not all of them have the optimal characteristics in terms of sensitivity to perform FOL detection in plant and seed material. A loop‐mediated isothermal amplification (LAMP) assay was developed based on the sequence‐characterized amplified region (SCAR) obtained in a previous rapid amplification of polymorphic DNA (RAPD) study. The LAMP assay has been validated according to the EPPO standard PM7/98. The LAMP assay was tested with lettuce seeds, soil and plant material, and can be used successfully to amplify DNA from each of these matrices. In seed lots artificially inoculated with FOL, the detection limit of the LAMP test was 0.004% infected seed.