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. 相似文献
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. 相似文献
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. 相似文献
The cellular immune response to tissue damage and infection requires the recruitment of blood leukocytes. This process is mediated through a classical multistep mechanism, which involves transient rolling on the endothelium and recognition of inflammation followed by extravasation. We have shown, by direct examination of blood monocyte functions in vivo, that a subset of monocytes patrols healthy tissues through long-range crawling on the resting endothelium. This patrolling behavior depended on the integrin LFA-1 and the chemokine receptor CX(3)CR1 and was required for rapid tissue invasion at the site of an infection by this "resident" monocyte population, which initiated an early immune response and differentiated into macrophages. 相似文献
Field monitoring and scenario-based modelling were used to assess exposure of small ditches in the UK to the herbicide sulfosulfuron following transport via field drains. A site in central England on a high pH, clay soil was treated with sulfosulfuron, and concentrations were monitored in the single drain outfall and in the receiving ditch 1 km downstream. Drainflow in the nine months following application totalled 283 mm. Pesticide lost in the first 12.5 mm of flow was 99% of the total loading to drains (0.5% of applied). Significant dilution was observed in the receiving ditch and quantifiable residues were only detected in one sample (0.06 microg litre(-1)). The MACRO model was evaluated against the field data with minimal calibration. The parameterisation over-estimated the importance of macropore flow at the site. As a consequence, the maximum concentration in drainflow (2.3 microg litre(-1)) and the total loading to drains (0.76 g) were over-estimated by factors of 2.4 and 5, respectively. MACRO was then used to simulate long-term fate of the herbicide for each of 20 environmental scenarios. Resulting estimates for concentrations of sulfosulfuron in a receiving ditch were weighted according to the prevalence of each scenario to produce a probability distribution of daily exposure. 相似文献
Aims: To assess the prevalence of obesity in adult dogs and cats presented to first-opinion veterinary clinics in the North Island of New Zealand for routine vaccination appointments, using electronic medical records.
Methods: Ten first-opinion veterinary clinics across the North Island of New Zealand provided electronic medical records for all routine vaccination appointments for adult (>1 year old) dogs and cats between 1 January 2011 and 30 June 2016. Animals with a body condition score (BCS) of 6 or 7 on a 9-point scale and 4 on a 5-point scale were classified as overweight; those with a BCS of 8 or 9 on a 9-point scale and 5 on a 5-point scale were classified as obese. A total of 106,144 records were available over the study period, of which 48,041 (45.2%) had both a recorded weight and BCS.
Results: Of the 24,247 records for dogs with both BCS and weight, 6,324 (26.1%) were classified as overweight, and 551 (2.3%) as obese. The prevalence of dogs classified as overweight or obese was highest in dogs aged between 5–13 years. The odds of desexed dogs being classified as overweight or obese was greater than the odds for intact dogs (OR=1.42 (95% CI=1.29–1.57), p<0.001) adjusting for the effects of age. Of the 23,794 records for cats with a recorded weight and BCS, 5,222 (21.9%) were classified as overweight, and 622 (2.6%) as obese. The prevalence of cats classified as overweight or obese was highest in cats aged between 5–11 years. The odds of desexed cats being classified as overweight or obese tended to be greater than the odds for intact cats (OR=1.14 (95% CI=0.98–1.31); p=0.075), adjusting for the effects of age.
Conclusions: Although there are limitations with using electronic medical records to estimate the prevalence of obesity in companion animal populations, the results highlight that a significant number of animals presenting for routine vaccination appointments were classified as overweight or obese.
Clinical Relevance: It is important for veterinarians to record both patient body condition and weight during routine preventative care appointments to allow accurate ongoing monitoring of trends in obesity at both the patient and population levels.