Cryopreservation of Ram Semen in Extenders Containing Soybean Lecithin as Cryoprotectant and Hyaluronic Acid as Antioxidant

A soybean lecithin‐based extender supplemented with hyaluronic acid (HA) was assayed for effectiveness to improve the quality of frozen–thawed ram semen. HA has not been tested yet in an extender containing soybean lecithin for freezing ram semen. Thus, the aim of this study was to analyse the effects of soybean lecithin at 1% or 1.5% along with HA at 0, 0.5 and 1 mg ml^‐1 in a Tris‐based extender on the motion characteristics, membrane integrity (HOST), viability, GSH peroxidase (GSH‐PX) activity, lipid peroxidation and acrosomal status after freezing–thawing. Semen was collected from four Mehraban rams during the breeding season and frozen in the six lecithin×HA extenders. The extender containing 1.5% lecithin supplemented with no HA yielded higher total motility (52.5%±1.6), viability (55.8%±1.6) and membrane integrity (44.5%±1.7), but the effects of the lecithin concentration did not reach signification. Linearity‐related parameters, ALH, BCF, lipid peroxidation, GSH‐PX activity, morphology and acrosomal status were not affected by the extender composition. In general, adding HA significantly decreased sperm velocity (1 mg ml^‐1 HA), total motility (only with 1.5% lecithin), viability (1 mg ml^‐1 HA for 1% lecithin; both concentrations for 1.5% lecithin) and membrane integrity. In conclusion, adding HA to the freezing extender supplemented with soybean lecithin failed to improve quality‐related variables in ram semen. Increasing the lecithin content could have a positive effect, but further studies are needed.     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

The toxic mixing zone of neutral and acidic river water: Acute aluminium toxicity in brown trout (Salmo trutta L.)

Mixing of acid river water containing aluminium (pH 5.1, Al 345 g.l^–1) with neutral water of a lake (pH 7.0, Al 73 g.l^–1) resulted in water (pH 6.4, Al 245 g.l^–1) with a pH (6.4) and Al concentration (245 g.l^–1) expected to have low toxicity to fish on the basis of current Al toxicity models. However, under semi-field conditions the freshly mixed water (a few sec. after mixing) proved to be highly toxic to brown trout. The fish were exposed to the water at different places along a 30 m channel. At the beginning of the channel acid and neutral water were continuously mixed; the mixed water left the channel after 340 sec. The cells of the gills showed a highly increased rate of cell death by apoptosis and necrosis. Intercellular spaces were enlarged, and many leucocytes penetrated in these spaces. Mucus release was stimulated to depletion. Plasma chloride levels were hardly affected. There was a clear gradient in the deleterious effects on the fish along the channel. The fish at the beginning of the channel (about 12 sec. after mixing of the water), were severely affected, whereas the fish kept at the end of the channel (340 sec. after mixing) were only mildly affected. In the natural situation fish will relatively quickly pass through a mixing zone. In our study we therefore focused on the effects on fish after a 60 min exposure to a mixing zone (5 sec after mixing), with subsequent recovery in a region downstream of the confluence and in neutral water with low Al. The recovery in the downstream area (at the end of the channel, i.e. 5 min after mixing) was clearly hampered when compared to the recovery in neutral water with low aluminium. Thus, a short exposure to the toxic mixing zone followed by a stay in water downstream of this zone, as may occur in nature, is detrimental to migrating trout. We conclude that freshly mixed acid and neutral water contain toxic components during the first seconds to minutes after mixing, that can not be explained by current models on aluminium toxicity.     

Superradiance in a torus magnetosphere around a black hole

The coalescence of a neutron star and a black hole in a binary system is believed to form a torus around a Kerr black hole. A similarly shaped magnetosphere then results from the remnant magnetic field of the neutron star. In the strong-field case, it contains a cavity for plasma waves located between the barrier of the gravitational potential and the surrounding torus. This cavity may be unstable to superradiance of electromagnetic waves. Superradiant amplification of such waves, initially excited by turbulence in the torus, should inflate into a bubble in a time as short as approximately 0.75 (1 percent/&cjs3539;epsilon&cjs3539;2)(M/7M middle dot in circle) seconds approximately 0.15 to 1.5 seconds, assuming an efficiency &cjs3539;epsilon&cjs3539;2 = 0.5 to 5 percent and a mass M = 7M middle dot in circle. These bubbles may burst and repeat, of possible relevance to intermittency in cosmological gamma-ray bursts. The model predicts gamma-ray bursts to be anticorrelated with their gravitational wave emissions.     

Salmon farming affects the fatty acid composition and taste of wild saithe Pollachius virens L.

There is an ongoing controversy on whether fish farming affects the quality of wild fish in fjords. In northern Norway, local people prefer not to eat saithe, Pollachius virens L., from areas in the vicinity of fish farms because they say the taste is inferior to saithe from other areas. To address this issue, saithe were collected in the vicinity of a salmon fish farm in a fjord in northern Norway and in two reference areas: one site 6 km away from the nearest fish farm in the same fjord, and the other in a fjord nearby with no fish farms. The objective of this study was to clarify whether the physiology or taste of saithe near fish farms differs from saithe in areas with no or limited influence from fish farms. The fish collected near farms were larger than those from control sites of the same age and had been eating pellets. Analysis of fatty acid composition of the fillet indicated that fish collected near the farms had fatty acid profiles that resembled that of pellets more than fish from reference sites. The results suggest that there were individual differences in the time spent near cages for saithe collected near the farm, and that saithe collected at the reference area in the same fjord had also been eating pellets, i.e. had visited at least one of the fish farms in the fjord. In a sensory test, the test panel found that saithe from the fjord without fish farms tasted better than saithe collected near the cages. The test panel found no clear differences in taste between saithe collected near the fish farm and saithe from the same fjord, but 6 km from the nearest fish farm.     

Effects of dietary cadmium on calcium homeostasis, Ca mobilization and bone deformities in Atlantic salmon (Salmo salar L.) parr

Atlantic salmon parr were reared for 4 months on experimental fish‐meal‐based diets supplemented with 0 (control), 0.5, 5, 25, 125 or 250 mg Cd kg^?1 feed to assess the effects of dietary Cd on active Ca uptake, mobilization of Ca from internal reservoirs, and development of bone malformations. The accumulation of dietary Cd in tissues was, in decreasing order, intestine > kidney >> gill > bone. No significant accumulation of Cd in the scales was observed. Strongest inhibition of ATP dependent Ca uptake (measured as Ca²⁺‐ and Na⁺/K⁺‐ATPase) was observed in the intestine of salmon fed 25 mg Cd kg^?1. This suppression in active intestinal Ca uptake did not lead to disturbed plasma Ca levels. Significant reduction of Ca from the scales in salmon fed 25 mg Cd kg^?1, indicates remobilization of Ca to maintain Ca homeostasis. At the end of the experiment no significant differences were observed in bone Ca levels, nor were any bone malformations observed in any of the dietary Cd‐exposed salmon. This indicates that bone as an endogenous Ca reservoir is spared compared with scales. It was concluded that dietary Cd‐induced disturbance of Ca homeostasis did not lead to bone deformities, even when Atlantic salmon were fed high amounts of cadmium. This indicates a low risk of spinal deformations being developed in Atlantic salmon sub‐chronically exposed to high Cd concentrations in the feed.     

Tissue lipid peroxidative responses in Atlantic salmon (Salmo salar L.) parr fed high levels of dietary copper and cadmium

Atlantic salmon (Salmo salar L.) parr were fed for one month on fish meal based diets supplemented with Cd (0, 0.7, or 204 mg Cd kg^–1 DW) or Cu (0, 34, or 691 mg Cu kg^–1 DW) to assess the effects of non-essential (Cd) and essential (Cu) dietary metals on lipid peroxidation and the oxidative defence system. Cadmium accumulated significantly in the liver, intestine, and kidney of 204 mg Cd kg^–1exposed fish compared to controls. Copper accumulated significantly in the intestine, kidney, and liver of fish exposed to 691 mg Cu kg^–1, and in the intestine of 34 mg Cu kg^–1 exposed fish. Tissue Cu accumulation significantly increased intestinal and hepatic lipid peroxidation (as seen from thiobarbituric acid reactive substances, TBARS, levels) and subsequently decreased intestinal -tocopherol levels and increased intestinal and hepatic selenium dependent glutathione peroxidase (SeGSH-Px) activity. Dietary Cd significantly reduced SeGSH-Px activity in the intestine and liver of 204 mg Cd kg^–1 exposed fish compared to controls. No significant increase in tissue TBARS or reduction of -tocopherol levels was observed in the intestine of fish exposed to dietary Cd, with exception of the highest exposure group (204 mg Cd kg^–1). Dietary Cu caused depletion of tissue Se and glutathione levels, however the reduced availability of GSH and Se did not seem to explain the differences in SeGSH-Px activity. Dietary Cu had a direct effect on lipid peroxidation at a relatively low concentration (34 mg Cu kg^–1). Cadmium indirectly affected tissue lipid peroxidation by damaging the oxidative defence system at the highest dietary concentration (204 mg Cd kg^–1).     

The Adverse Effect of Phytoestrogens on the Synthesis and Secretion of Ovarian Oxytocin in Cattle

The current investigations were undertaken to study the mechanism of the adverse effect of phytoestrogens on the function of bovine granulosa (follicles >1< cm in diameter) and luteal cells from day 1–5, 6–10, 11–15, 16–19 of the oestrous cycle. The cells were incubated with genistein, daidzein or coumestrol (each at the dose of 1 × 10^?6 m ). The viability and secretion of estradiol (E2), progesterone (P4) and oxytocin (OT) were measured after 72 h of incubation. Moreover, the expression of mRNA for neurophysin‐I/OT (NP‐I/OT; precursor of OT) and peptidyl‐glycine‐α‐amidating monooxygenase (PGA, an enzyme responsible for post‐translational OT synthesis) was determined after 8 h of treatment. None of the phytoestrogens used affected the viability of cells except for coumestrol. The increased secretion of E2 and P4 was only obtained by coumestrol (p < 0.05) from granulosa cells from follicles <1 cm in diameter and decreased from luteal cells on days 11–15 of the oestrous cycle, respectively. All three phytoestrogens stimulated (p < 0.05) OT secretion from granulosa and luteal cells in all stages of the oestrous cycle and the expression of NP‐I/OT mRNA in the both types of cells. The expression of mRNA for PGA was stimulated (p < 0.05) by daidzein and coumestrol in granulosa cells, and by genistein and coumestrol in luteal cells. In conclusion, our results demonstrate that these phytoestrogens can impair the ovary function in cattle by adversely affecting the synthesis of OT in follicles and in corpus luteum. However, their influence on the ovarian steroids secretion was less evident.     

Effects of herbivory on the reproductive effort of 4 prairie perennials

Background  

Herbivory can affect every aspect of a plant's life. Damaged individuals may show decreased survivorship and reproductive output. Additionally, specific plant species (legumes) and tissues (flowers) are often selectively targeted by herbivores, like deer. These types of herbivory influence a plant's growth and abundance. The objective of this study was to identify the effects of leaf and meristem removal (simulated herbivory within an exclosure) on fruit and flower production in four species (Rhus glabra, Rosa arkansana, Lathyrus venosus, and Phlox pilosa) which are known targets of deer herbivory.