Follicullar Development and Embryo Recovery Following 3 versus 8 FSH Injections in Heifers

Ovarian follicular dynamics and embryo yield were studied during 2 different FSH regimens for superovulation of cattle. Twenty heifers were given intramuscular injections of FSH (total of 35 mg NIH) either once daily for 3 days (Group 3×1) or twice daily for 4 days (Group 4×2). At 72 h after the first FSH injection, each animal was injected with 0.75 mg cloprostenol. Inseminations were performed at 12 h and 24 h after the onset of heat. Transrectal ultrasonography was performed on the day of the first FSH injection, the day of cloprostenol injection, the day of insemination and finally on the day of embryo recovery (day 6 or 7 after heat). The numbers of small (2–4 mm), medium (5–9 mm) and large (>10 mm) size follicles were recorded. The total number of corpora lutea, eggs and transferable embryos were recorded on the day of embryo recovery. No differences were found between the 2 groups in either of the parameters studied (p>0.05). It can be concluded that treatment with this FSH preparation once daily for 3 days gives a folliculogenic and superovulatory response similar to a treatment regimen where it is given twice daily for 4 days.     

Relationship between blood iron parameters and trichinellosis in swine

Reduced levels of total iron binding capacity and unsaturated iron binding capacity were observed in the blood of trichinous and iron-injected trichinous pigs. No change was observed in their serum iron and saturation concentration levels. Also, reduced iron concentration levels were observed in the livers of trichinous pigs, while increased iron concentration levels were observed in the spleens of trichinous pigs and the livers and spleens of iron-injected pigs. No difference was found with regard to weight gains, number of larvae per gram of tissues, or histologic characteristics of 'nurse cells'.     

Estimating adrenal cortical function in dogs with ACTH.

The peripheral blood response to intramuscular injection of 10 units ACTH in dogs was investigated because no experimental evidence for the standardization of this procedure for clinical use was available. Following the injection of ACTH in sodium chloride solution, neutrophilia, monocytosis, eosinopenia, and lymphopenia occurred. With the exception of eosinopenia, the greatest change in the concentration of each cell type in peripheral blood occurred between 2 and 4 hours post injection. The maximum change in eosinophil numbers occurred between 4 and 6 hours post injection. When all cell types were considered, 4 hours post injection was the most suitable time to measure the cellular response in peripheral blood in dogs which respond to ACTH. The data indicate that change in the ratio of neutrophils to lymphocytes (N/L) prior to and at 2 to 4 hours after ACTH injection in normal dogs was a sensitive index of response and occured sooner than eosinopenia. The extent of change in the N/L ratio was such that accuracy in interpretation could be obtained by counting as few as 40 cells.     

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.     

In Vitro Assessment of the Equine Hoof Wall Strains in Flat Weight Bearing and After Heel Elevation

Horseshoeing is a common practice, but effects on the hoof wall are poorly understood. Strain gauges were used to document and compare hoof behavior in vitro during flat weight bearing and after artificial heel elevation. Ten front limbs of Thoroughbred race horses, shod with conventional flat shoes, were used. Eight strain gauges were symmetrically distributed around the toe, quarters, and heels. Each limb was mounted to a testing machine (Kratos K5002; Kratos Dynamômetros, Ltda., Cotia-SP-Brazil) and subjected to a load equivalent to 30% of the donor's body weight. Strains (μ) were acquired by means of a computerized system and the results compared using Friedman and Wilcoxon statistical tests. There was greater strain variation when the heels were elevated. Compression predominated during flat weight bearing, with a tendency to horizontal traction after heel elevation. The changes in strain caused by heel elevation were not always symmetrical. Elevation of the heels tensed the toe and the medial quarter horizontally, increased load at the posterior portion of the hoof capsule, and hindered its expansion.     

Embryo Transplantation in Cattle: Non-Surgical Recovery of Embryos From Repeat Breeders

Fourteen true repeat breeders with entirely normal oestrous cyclicity more than 1 year after calving and 14 control donor cows were superovulated with PMSG (2000 i.u.) and flushed non-surgically 6–8 days after the superovulatory heat. The superovulatory response was identical for the 2 groups such as assessed by the number of corpora lutea (9.4 ± 1.8 C.L. per repeat breeder and 9.1 ± 1.5 per control cow), occurrence of ovarian overstimulation (polycysts), presence of a non-countable amount of corpora lutea, negative outcome of the flushings and the number of recovered embryos (5.8 ± 1.0 embryos per repeat breeder and 6.0 ± 1.8 embryos per control cow). The most pronounced difference between the 2 categories of animals was related to the fertilization rate of embryos. In the repeat breeder group only 2.4 embryos per cow or 41 % were fertilized, whereas the control animals attained a fertilization rate of 4.9 embryos or 82 %. Since most factors liable to interfere with the fertilization process were identical for both groups (age, breed, nutritional and management conditions, semen quality, dose, AI-technician e.g.), it is believed that intraovarian, follicular, or follicular-dynamic conditions were responsible for producing a high proportion of non-fertilizable oocytes.