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.     

Two-year development of southern pine seedlings and associated vegetation following spray-and-burn site preparation with imazapyr alone or in mixture with other herbicides

Eight herbicide combinations containing imazapyr alone or in mixture with glyphosate, triclopyr, or picloram+2,4-D were applied July–August 1991 at each of three upland sites in the southeastern U.S., with or without a subsequent broadcast burn in September. Seedlings of loblolly (Pinus taeda L.) or slash (P. elliottii Engelm. var. elliottii) pine were planted that winter. Second-year (August 1993) vegetation responses indicate that broadcast burning, in combination with herbicides, was associated with increased abundances of grasses, forbs, and shrubs and decreased abundances of vines and hardwood trees. In the presence of burning, imazapyr-in-mixture treatments were associated with a high abundance of grass (53% cover) and a mean size of pines less than that of seedlings growing in imazapyr-alone treatments. In the absence of burning, imazapyr-alone treatments were associated with a high abundance of blackberry (21% cover), an average height of hardwoods (131 cm) exceeding that of pines (71 cm), and a mean size of pines less than that of trees growing in imazapyr-in-mixture treatments. Hardwood height in the absence of burning was less when imazapyr was combined with glyphosate (75 cm) than with triclopyr (102 cm). Regardless of the presence or absence of burning, size of pines and abundance of shrubs and hardwoods did not differ between imazapyr rates of 0.56 and 0.84 kg acid equivalent/ha or among herbicides applied in mixture with imazapyr.     

THE EFFECT OF CUTTING HEIGHT AND NITROGEN LEVEL ON THE YIELD, IN VITRO DIGESTIBILITY AND CHEMICAL COMPOSITION OF ITALIAN RYEGRASS SWARDS

Three cutting heights, 2±5, 7±6 and 12±7 cm (1, 2±5 and 5 in.) and three levels of fertilizer N, 168, 280 and 392 kg N/ha (150, 250 and 350 Ib N/ac) were imposed on a sward of Italian ryegrass (Lolium multiflorum) cv. Irish. Lowering the cutting height and increasing the level of applied N increased the yield of herhage DM. Increasing the level of applied N had a greater effect on the chemical composition of the herbage than altering the cutting height     

Temporal and spatial dynamics of insecticide resistance in Myzus persicae (Hemiptera: Aphididae)

Advances in understanding insecticide resistance in the peach-potato aphid, Myzus persicae (Sulzer), at the genotypic, biochemical and molecular levels have led to rapid and precise methods for the detection of several resistance mechanisms (elevated carboxylesterase, modified acetylcholinesterase or MACE, and knockdown resistance or kdr) in individual insects, and for monitoring their frequencies over space and time. This paper summarises the results of two long-term surveys of resistance dynamics in M persicae in England, based on samples collected directly from field and glasshouse crops or from four 12.2-m suction traps. The study showed marked fluctuations in resistance frequencies that probably reflect the counteracting forces of selection imposed by insecticides for aphids possessing more copies of esterase resistance genes, and selection against these forms when insecticide use is relaxed. There is growing evidence that several different resistance mechanisms in M persicae have associated fitness costs. In the case of esterase and MACE, these costs are apparently strong enough to effect a decline in resistance frequency over winter, and a more prolonged decline over successive cropping seasons when aphid numbers are insufficient to trigger intensive chemical applications. Changes in the overall frequency of resistance genotypes may also be influenced by the predominance of year-round parthenogenesis in M persicae in the UK, leading to non-random associations between mechanisms and selection operating on clonal lineages rather than individual genotypes.