Spermatic and oxidative profile of domestic cat (Felis catus) epididymal sperm subjected to different cooling times (24, 48 and 72 hours)

Cooling stored epididymal samples for several days allows facilities to transport and process genetic material post‐mortem. Improvements to this practice allow the preservation of sperm from domestic cats, which are the ideal study model for wild felids. However, the modifications in spermatic features and the oxidative profile are not fully understood in cats. This information is necessary for the development of biotechniques, such as new extenders for cryopreservation. Therefore, the purpose of this study was to evaluate the spermatic and oxidative profile in samples from the epididymal cauda of domestic cats cooled at 5°C for 24, 48 and 72 hr. Spermatozoa were collected from the epididymis cauda. Evaluations consisted of computer‐assisted sperm analysis (CASA), plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, sperm DNA integrity (toluidine blue), mitochondrial activity (3′3 diaminobenzidine), activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD), measurement of lipid peroxidation (TBARS) and protein oxidation. A decrease in sperm motility parameters was observed after 72 hr of cooling (i.e. total and progressive) with a higher percentage of minor (37.7 ± 6.3%) and total defects (53.4 ± 6.3%). Additionally, a decrease in high mitochondrial activity (Class I: 16.6 ± 2.2%) occurred after 72 hr. The decrease in motility rates after a long cooling time probably was caused by the increase in sperm abnormalities. A long cooling time causes cold shock and mitochondrial exhaustion, but there was no observed change with the oxidative stress condition. Therefore, cat epididymal sperm stored at 5°C appear to maintain a high quality for up to 48 hr of cooling time.     

A fast,low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species

Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high‐precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For this purpose, we used six animals of each specie (n = 30), in reproductive age. Sperm was collected by different methods according to the particularities of each species, and such samples were divided into two aliquots: a sperm sample was kept at 5°C (considered as intact sperm DNA), and the remaining samples were submitted to the induction of DNA fragmentation by exposure to ultraviolet light for 4 hr. Samples were then mixed with the intact sample to obtain known and progressive proportions of sperm with fragmented DNA (0%, 25%, 50%, 75% and 100%). Semen smears were performed and subjected to staining with TB. Blue‐stained spermatozoa were considered to have DNA fragmentation. We observed high linear regression coefficients between the expected proportion of damaged DNA and the results of TB for dog, cat, ram, bull and stallion samples. In conclusion, TB stain was considered a fast and effective technique for the study of spermatozoa DNA in several species.     

Long-term tillage and rotation effects on soil microbial biomass,carbon and nitrogen

Summary Three mollisols, typical of the Palouse winter wheat region of eastern Washington and northern Idaho, were analyzed for microbial biomass, total C and total N after 10 years of combined tillage and rotation treatments. Treatments included till, no-till and three different cereal-legume rotations. All crop phases in each rotation were sampled in the same year. Microbial biomass was monitored from April to October, using a respiratory-response method. Microbial biomass, total C and total N were highest under no-till surface soils (0–5 cm), with minimal differences for tillage or depth below 5 cm. Microbial biomass differences among rotations were not large, owing to the relative homogeneity of the treatments. A rotation with two legume crops had the highest total C and N. Microbial biomass was significantly higher in no-till surface soils where the current crop had been preceded by a high-residue crop. The opposite was true for the tilled plots. There was little change in microbial biomass over the seasons until October, when fresh crop residues and rains had a strong stimulatory effect. The seasonal pattern of biomass in no-till surface soils reflected the dry summer/winter rainfall climate of the region. The results of this study show that numerous factors affect soil microbial biomass and that cropping history and seasonal changes must be taken into account when microbial biomass data are compared.Scientific paper no. 7634     

Sectional notes

American Journal of Potato Research -     

The effects of several supplementation frequencies on forage use and the performance of beef cattle consuming dormant tallgrass prairie forage

Two experiments were conducted to quantify the impact on forage use and performance of varying supplementation frequency of cattle consuming forage diets across a range of frequencies. In both experiments, a common supplement was used that contained a relatively high concentration of CP (43%) and was fed at the following frequencies: 1) 2 d/wk; 2) 3 d/wk; 3) 5 d/wk; and 4) 7 d/wk. In Exp. 1, 120 Hereford x Angus cows (BW = 537 kg) grazing winter tallgrass-prairie range were supplemented at the various frequencies from December 7 until calving (average calving date = 3/7/99). All treatments provided the same quantity of supplement on a weekly basis (12.74 kg, as-fed) but divided the amount delivered on a given day equally among the number of supplementation events for that treatment. Less BW was lost from December 7 through calving (linear effect, P = 0.02) as frequency of supplementation increased, but the magnitude of difference in weight change was relatively small. Body condition responded similarly through early February (linear effect, P = 0.02), although treatment effects were not as distinct at calving (cubic effect, P = 0.11). In Exp. 2, 16 ruminally fistulated Hereford x Angus steers (BW = 257 kg) were blocked by weight and assigned to one of the four frequencies of supplementation. Steers were offered tallgrass prairie hay (73.5% NDF, 4.8% CP) ad libitum and were supplemented at a rate (relative to BW) similar to that of the cows in Exp. 1. Increasing frequency of supplementation increased (linear effect, P < or = 0.02) forage OM intake, OM and NDF digestion, and digestible OM intake. However, the most prominent differences in forage OM intake tended (cubic effect, P = 0.07) to occur with the two extreme frequencies of supplementation. In conclusion, forage use was improved with an increased frequency of supplementation, but the impact on performance is not likely to be large unless extreme differences in frequency occur.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Soil microbial, fungal, and nematode responses to soil fumigation and cover crops under potato production

Sodium N-methyldithiocarbamate (metam sodium) and 1,3 dichloropropene are widely used in potato production for the control of soil-borne pathogens, weeds, and plant parasitic nematodes that reduce crop yield and quality. Soil fumigation with metam sodium has been shown in microcosm studies to significantly reduce soil microbial populations and important soil processes such as C and N mineralization. However, few published data report the impact of metam sodium on microbial populations and activities in potato production systems under field conditions. Fall-planted white mustard (Brassica hirta) and sudangrass (Sorghum sudanense) cover crops may serve as an alternative to soil fumigation. The effect of metam sodium and cover crops was determined on soil microbial populations, soil-borne pathogens (Verticillium dahliae, Pythium spp., and Fusarium spp.), free-living and plant-parasitic nematodes, and C and N mineralization potentials under potato production on five soil types in the Columbia Basin of Eastern Washington. Microbial biomass C was 8–23% greater in cover crop treatments compared to those fumigated with metam sodium among the soil types tested. Replacing fumigation with cover crops did not significantly affect C or N mineralization potentials. Cumulative N mineralized over a 49-day laboratory incubation averaged 18 mg NO₃-N kg⁻¹ soil across all soil types and treatments. There was a general trend for N mineralized from fumigated treatments to be lower than cover-cropped treatments. Soil fungal populations and free-living nematode levels were significantly lowered in fumigated field trials compared to cover-cropped treatments. Fumigation among the five soil types significantly reduced Pythium spp. by 97%, Fusarium spp. by 84%, and V. dahliae by 56% compared to the mustard cover crop treatment. The percentage of bacteria and fungi surviving fumigation was greater for fine- than coarse-textured soils, suggesting physical protection of organisms within the soil matrix or a reduced penetration and distribution of the fumigants. This suggests the potential need for a higher rate of fumigant to be used in fine-textured soils to obtain comparable reductions in soil-borne pathogens.     

Earth tides can trigger shallow thrust fault earthquakes

We show a correlation between the occurrence of shallow thrust earthquakes and the occurrence of the strongest tides. The rate of earthquakes varies from the background rate by a factor of 3 with the tidal stress. The highest correlation is found when we assume a coefficient of friction of mu = 0.4 for the crust, although we see good correlation for mu between 0.2 and 0.6. Our results quantify the effect of applied stress on earthquake triggering, a key factor in understanding earthquake nucleation and cascades whereby one earthquake triggers others.