The Effect of Oxidative Stress on Thawed Bulk‐Sorted Red Deer Sperm

The aims of this study were to assess the effects of the sex‐sorting process on post‐thaw sperm quality as well as on induced oxidative stress damage (H₂O₂ 0 mm  = H000; H₂O₂ 50 mm  = H050; H₂O₂ 100 mm  = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non‐sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H₂O₂ increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm‐sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non‐sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.     

Selection of red deer spermatozoa with different cryoresistance using density gradients

The objective of sperm selection media is selecting the best spermatozoa and to remove seminal plasma and diluent for using them in assisted reproductive techniques. It is known that individuals show different cryoresistance in response to the same freezing procedure. Our hypothesis was that the efficacy of selection media could be dissimilar for samples with different sperm quality after thawing. Epididymal sperm samples from mature Iberian red deer were collected and frozen. Males were classified as with high post‐thaw sperm quality when sperm motility (SM) ≥ 70%, or as with low post‐thaw sperm quality when SM ≤ 69%. Samples were centrifuged using the following density gradients (DG): Percoll^®, Puresperm^® and Bovipure^?, and several functional sperm parameters were assessed after sperm selecting and washing. Males classified with high sperm quality had higher post‐thawing values (p > .05) for all parameters evaluated, except for linearity index, than those categorized as low sperm quality. After selection, some sperm characteristics improved (viability, apoptosis and mitochondrial activity) for both groups, showing the males with high sperm quality higher values in all sperm parameters except for kinematic traits and DNA fragmentation index (%DFI), regardless of DG. Bovipure^? yield lower values of sperm motility, viability, apoptosis and mitochondrial activity in relation to Percoll^® and Puresperm^® considering both quality groups. There was an interaction between the type of DG and sperm quality group for sperm viability (p = .040) and apoptosis (p = .003). Thus, Percoll^® selected less live and more apoptotic spermatozoa than Puresperm^® and Bovipure^? for males with low sperm quality. In conclusion, the DG are more efficient selecting spermatozoa from samples with high sperm quality, acting differently depending on initial sperm quality.     

Effects of two large doses of equine recombinant growth hormone on clinical, haematological and serum biochemical variables in adult horses

Objective To evaluate the clinical, haematological, and serum biochemical effects of two large doses of recombinant equine growth hormone.
Design Duplicated Latin square.
Sample population
Three Thoroughbred and three Standardbred mares aged between 12 and 17 years.
Procedure Two horses were randomly assigned into one of three groups. On each of three successive days, each horse pair received one of two dosages of growth hormone or a saline placebo so that by the end of the experiment all three horse pairs had received both dosages and the saline placebo. Dose rates selected were 50 μg/kg, and 100 μg/kg. A clinical examination was performed and a venous blood sample drawn for a complete blood count and serum biochemical analysis before administration of growth hormone and at 1, 2, 3, 4, 6, 8 and 24 h after injection. Data were analysed by a repeated measures analysis of variance assessing the effects of dose and time.
Results There was an effect of time on a number of clinical, haematological, and serum biochemical variables. There were significant effects of growth hormone on heart rate and serum glucose concentration but values for both variables remained within the reference range.
Conclusion The results of the present study suggest that equine recombinant growth hormone has a wide margin of safety and show that the single administration of up to five times the recommended dose rate has no significant effects on clinical, haematological, or serum biochemical variables.     

Luteolytic Effect of Prostaglandin F2α on Bovine Corpus Luteum Depends on Cell Composition and Contact

Prostaglandin F_2α (PGF_2α) is a main luteolytic factor in vivo; however, its direct luteolytic influence on steroidogenic cells of bovine corpus luteum (CL) is controversial and not fully understood. The aim of the study was to clarify PGF_2α action on bovine CL in different in vivo and in vitro conditions and to examine whether the contact among all main types of CL cells is necessary for luteolytic PGF_2α action. In experiment 1, the bovine CL (day 15 of the oestrous cycle) was perfused using in vivo microdialysis system with dinoprost (an analogue of PGF_2α) for 0.5 h. Dinoprost caused a short‐time increase in progesterone (P4), whose concentration decreased thereafter (at 6‐, 10‐, 12‐ and 24‐h after treatment). In experiment 2, the direct effect of PGF_2α on P4 accumulation in CL steroidogenic cells cultured in monolayer (day 15 of the cycle) was determined. PGF_2α after 24 h of incubation increased P4 accumulation in steroidogenic CL cells. In experiment 3 steroidogenic, endothelial CL and immune cells (day 15 of the cycle) were incubated with PGF_2α in cocultures for 24 h in glass tubes and the levels of P4, stable metabolites of nitric oxide (NO) and leukotriene (LT) C₄ were determined. Although PGF_2α treatment increased P4 secretion in homogeneous steroidogenic CL cell culture, the decrease in P4 secretion in cocultures of all types of CL cells was observed. The secretion of NO and LTC₄ increased after the treatment of PGF_2α both in pure cultures of CL cells and in cocultures. The interactions between endothelial and immune cells with steroidogenic CL cells are needed for luteolytic PGF_2α action within the bovine CL. Our results indicate that the cell coculture model, including the main types of CL cells, is the most approximate to study PGF_2α role in vitro.     

不同Rht基因在冬小麦育种中的利弊分析

利用携带不同的Ｒｈｔ矮秆基因的６个冬小麦近等基因系,进行了两年试验,研究了不同矮秆基因在育种中的利用价值,结果表明,Ｒｈｔ１和Ｒｈｔ２两个半矮秆基因提高了单株（或单穗）粒数和粒重,经济系数和倒二叶面积。产Ｒｈｔ３和高秆系显著降低了千粒重,Ｒｈｔ３矮秆基因对单株粒数,单株产量和穗粒数均负向作用,显著提高了千粒重;Ｒｈｔ１＋Ｒｈｔ２较高秆系和Ｒｈｔ２＋Ｒｈｔ３系比较显著提高了经济系数,Ｒｈｔ２＋Ｒｈ５     

Effects of sheep stocking on the plant community and agricultural characteristics of upland Anthoxanthum odoratum–Geranium sylvaticum meadow in northern England

In the UK, upland hay meadows (Anthoxanthum odoratum–Geranium sylvaticum grassland) with high plant biodiversity are rare and confined to submontane areas of northern England. We report results from a 5‐year experiment to test suggestions that recent biodiversity declines were attributable to increased sheep‐stocking density and a longer spring stocking period, thus delaying the shut up date for the growth of the hay crop. Longer stocking periods and higher stocking densities decreased the forage mass at 8 July, but they increased herbage N content and digestibility, reduced plant species diversity and reduced populations and seed production of Rhinanthus minor. Compared with unstocked swards, the similarity of the vegetation to Anthoxanthum odoratum–Geranium sylvaticum grassland was reduced by 16·9% when stocking with sheep continued until 27 May each year, and by 8·3% when sward heights were maintained at 3 cm compared with 5 cm. Increased mean sward height and height of R. minor were positively correlated with accumulated temperatures. Results support suggestions that recent reductions in the nature value of these grasslands might be a consequence of high stocking densities persisting until later in the spring, carried out during a 1‐year period with warmer temperatures.