Presence of Maedi Visna Virus (MVV)‐Proviral DNA in the Genital Tissues of Naturally Infected Ewes

Maedi Visna virus (MVV) causes progressive degenerative inflammatory disease in multiple organs including the lungs (pneumonia, ‘maedi’), mammary gland, joints and nervous system (meningoencephalomyelitis, ‘visna’) in sheep. Maedi Visna Virus has been detected in macrophages of several tissues and epithelial cells in vivo: bone marrow, cells of the central nervous system, lung and bronchial tissues, milk epithelial cells recovered from milk samples and epithelial cells of mammary tissue. However, the presence of MVV in the genital tracts of naturally infected ewes has not previously been studied. The aim of this study was to use nested‐PCR, targeting the gag gene, to determine whether genital tissues (ovaries, oviducts and uterus) from 83 ewes originating from various breeding herds in the South‐East of France were positive for MVV‐proviral DNA. Peripheral blood mononuclear cells (PBMC) tested positive for MVV‐proviral DNA, using nested‐PCR analysis, in 57.8% of ewes (48/83). The provirus was also identified in 47% (78/166) of the ovaries, 38.6% (64/166) of the oviducts and 45.8% (38/83) of the uteri sampled. These findings clearly demonstrate, for the first time, that tissue samples from the genital tract of ewes (ovary, oviduct and uterus) can be infected with MVV. This suggests that there is a risk of vertical and/or horizontal transmission of MVV during embryo transfer from embryos produced in vivo or in vitro.     

Measurement of Parity Nonconservation and an Anapole Moment in Cesium

The amplitude of the parity-nonconserving transition between the 6S and 7S states of cesium was precisely measured with the use of a spin-polarized atomic beam. This measurement gives Im(E1pnc)/beta = -1.5935(56) millivolts per centimeter and provides an improved test of the standard model at low energy, including a value for the S parameter of -1.3(3)exp (11)theory. The nuclear spin-dependent contribution was 0.077(11) millivolts per centimeter; this contribution is a manifestation of parity violation in atomic nuclei and is a measurement of the long-sought anapole moment.     

In Vitro Evaluation of Antioxidant Potential of the Invasive Seagrass Halophila stipulacea

Marine organisms with fast growth rates and great biological adaptive capacity might have biotechnological interests, since ecological competitiveness might rely on enhanced physiological or biochemical processes’ capability promoting protection, defense, or repair intracellular damages. The invasive seagrass Halophila stipulacea, a non-indigenous species widespread in the Mediterranean Sea, belongs to this category. This is the premise to investigate the biotechnological interest of this species. In this study, we investigated the antioxidant activity in vitro, both in scavenging reactive oxygen species and in repairing damages from oxidative stress on the fibroblast human cell line WI-38. Together with the biochemical analysis, the antioxidant activity was characterized by the study of the expression of oxidative stress gene in WI-38 cells in presence or absence of the H. stipulacea extract. Concomitantly, the pigment pool of the extracts, as well as their macromolecular composition was characterized. This study was done separately on mature and young leaves. Results indicated that mature leaves exerted a great activity in scavenging reactive oxygen species and repairing damages from oxidative stress in the WI-38 cell line. This activity was paralleled to an enhanced carotenoids content in the mature leaf extracts and a higher carbohydrate contribution to organic matter. Our results suggest a potential of the old leaves of H. stipulacea as oxidative stress damage protecting or repair agents in fibroblast cell lines. This study paves the way to transmute the invasive H. stipulacea environmental threat in goods for human health.     

Comparison of Membrane‐Permeant Fluorescent Probes for Sperm Viability Assessment in the Ram

This study was designed to compare the effectiveness and usability of four permeant fluorochromes (CFDA; SYBR‐14; Hoechst‐33342; and acridine orange), combined with propidium iodide to assess sperm membrane integrity. Three different experiments were conducted. The first trial was designed to study the optimal dye concentration and minimum incubation time required to achieve optimum fluorescence intensities and contrast for each fluorochrome combination using ram fresh semen samples. Both SYBR‐14 and acridine orange allowed a direct assessment of sperm membrane integrity, without the need of incubating samples, whereas a minimum of 4 and 6 min of incubation at 37°C was necessary to achieve optimum fluorescence intensities in the CFDA and Hoechst groups, respectively. In the second trial, fresh semen samples were mixed with different volumes of membrane‐affected sperm (semen treated with three cycles of freezing to ?20°C and thawing at room temperature) to produce semen samples with known proportions of damaged spermatozoa. The results were compared with the theoretical values predicted on the basis of the estimations made on fresh and frozen samples. The proportions of damaged sperm in each sample determined using the four fluorochrome combinations agreed with the predicted theoretical values, with the acridine orange/propidium iodide providing the best adjustment. The third experiment was performed to compare the results of sperm membrane integrity using the four fluorochrome combinations. The proportions of plasmalemma‐intact sperm determined by acridine orange and SYBR‐14 were greater (p < 0.0001) than the proportions of intact sperm determined by CFDA and Hoechst stains. It was concluded that the most efficient combinations to be used in ram sperm were AO/PI and SYBR/PI because it allowed a direct assessment of sperm viability without the need to incubate samples and obtaining reliable results.     

The Effects of Sample Size on the Outcome of Ovarian Tissue Cryopreservation

Cryopreservation of ovarian tissue is known to affect follicular survival. Several variables may be responsible for this. Little attention has focused on the effect of the size of the fragment to be cryopreserved. This study was conducted to assess the effect of the size of the tissue on follicular histology after freezing with 1,2-propanediol. Histological evaluations were performed of control and cryopreserved tissue. Fragments were cut 10 × 3 × 2 mm³ (2 mm group) or 10 × 3 × 4 mm³ (4 mm group). Percentages of normal follicles in control fragments cut into 2 and 4 mm slices were 56% and 34%, respectively. The relative risks to obtain normal follicles in the 2 mm and the 4 mm fragments after cryopreservation were 0.63 and 0.47, respectively. Freezing reduced follicle survival to a significantly greater extent in the larger tissue fragments. There is an increased risk of damage to primary and primordial follicles, when the tissue slices are cut with all dimensions larger than 2 mm.     

Association with TGF‐β1 Gene Polymorphisms and Reproductive Performance of Large White Pig

As a multifunctional cytokine, transforming growth factor‐beta1 (TGF‐β1) was detected in the utero‐placental interface during early pregnancy in the pig and believed to enhance trophoblast attachment to the endometrium. In this experiment, we selected TGF‐β1 as the candidate gene affecting litter size in pigs. Four polymorphic loci of TGF‐β1 gene were found by PCR‐SSCP (single‐strand conformation polymorphism) in Large white sows (n = 567): C→T mutation at 33nt in the intron 4; G→A mutation at 179nt in the intron 6; C→T mutation at 1043nt in the intron 6; GG→AA linkage mutations at 2490nt and 2494nt respectively. We haplotyped these SNPs as: CGCAA (denote as P) and TATGG (denote as K). The effects of three haplotypic combinations (HCs) of PP, PK and KK on litter sizes were investigated by a linear model. It was found that for the first parity litters, the least squares mean for total number born (TNB) of KK was 1.02 piglets/litter, higher than that of PK (p < 0.05), 0.49 piglets/litter higher than that of PP (p > 0.1). There were no significant differences between HCs on the second parity. The result indicated that KK HCs was significantly associated with pig litter size.     

The Influence of Powdered Coconut Water (ACP‐318®) in In Vitro Maturation of Canine Oocytes

The objective of this study was to determine the influence of powdered coconut water (ACP‐318^®) diluted in high glucose (11.0 mm ) TCM199 in the achievement of nuclear in vitro maturation (IVM) of canine oocytes. Cumulus oocyte complexes (COCs) (n = 632) were randomly allocated into three experimental groups named as group 1 (control group), group 2 (5% powdered coconut water) and group 3 (10% powdered coconut water). The percentage of meiotic resumption (MR) (GVBD to MII) was 39.1% (81/207), 50.2% (108/215) and 46.6% (98/210) for groups 1, 2 and 3 respectively (p < 0.05). There were no differences in MR rates among groups 2 and 3. The medium with ACP‐318^® slightly enhanced the nuclear maturation of canine oocytes when a comparison was established with rates of maturation exhibited by oocytes in the experimental group 1 without ACP‐318^® (p < 0.05). The results suggest that oocytes’ nuclear morphology integrity and meiosis achievement were positively influenced when exposed to high glucose TCM199 supplemented with 5% powdered coconut water. Further investigation must be performed for a better understanding of powdered coconut water influence in cellular events during IVM of dog oocytes.     

Replacing coniferous monocultures with mixed-species production stands: An assessment of the potential benefits for forest biodiversity in northern Europe

Conifer dominated plantations in central and northern Europe are associated with relatively low ecological values, and in some cases, may be vulnerable to disturbances caused by anthropogenic climate change. This has prompted the consideration of alternative tree species compositions for use in production forestry in this region. Here we evaluate the likely biodiversity costs and benefits of supplanting Norway spruce (Picea abies) monocultures with polycultures of spruce and birch (Betula spp.) in southern Sweden. This polyculture alternative has previously been evaluated in terms of economic, recreational, and silvicultural benefits. By also assessing the ecological implications we fill a gap in our understanding of the range of socio-ecological benefits that can be achieved from a single polyculture alternative. We project likely broad scale changes to species richness and abundance within production stands for five taxonomic groups including ground vegetation, tree-living bryophytes, lichens, saproxylic beetles, and birds. Our research leads us to three key findings. First, the replacement of spruce monocultures with spruce–birch polycultures in the managed forest landscapes of southern Sweden can be expected to result in an increase in biological diversity for most but not all taxa assessed, but it is unlikely to improve conditions for many red-listed forest species. Second, modification of other aspects of forest management (i.e. rotation length, dead wood and green tree retention, thinning regimes) is likely to contribute to further biodiversity gains using spruce–birch polycultures than spruce monocultures. Third, the paucity of empirical research which directly compares the biodiversity of different types of managed production stands, limits the extent to which policy relevant conclusions can be extracted from the scientific literature. We discuss the wider implications of our findings, which indicate that some climate change adaptation strategies, such as risk-spreading, can be readily integrated with the economic, environmental and social goals of multi-use forestry.