Assessment of ovarian follicular dynamics and folliculogenesis associated endocrine profiles following gonadotropin stimulation in the bonnet monkey.

We evaluated ovarian follicular dynamics in bonnet monkeys by employing trans-abdominal ultrasonography. Following the administration of human follicle stimulating hormone (hFSH) and/or human menopausal gonadotropin (hMG), multiple follicular development was assessed and their numbers, size and growth profiles were monitored. The ultrasonograms showed that the follicular antrum appeared distinctly anechoic with well-defined hyperechoic borders. Depending on the type, quantity (12.5-25IU), and duration (6-9days) of hormones administered, the number of developing follicles was 2-12 per ovary with their lowest diameter being 2mm. With continued hormone administration, their numbers and diameters increased; which were more pronounced in animals administered with hFSH than with hMG, with follicles of 6-8mm. Interestingly, human chorionic gonadotropin (hCG) injection (2000-3000IU), when follicles acquiring >6-8mm sizes, induced the maximum expansion of antral follicles with sizes reaching up to 14mm. On days 3-5 post-hCG, the ultrasonograms showed loosely demarcated multiple hypoechoic structures and well-demarcated hyperechoic structures with anechoic/hypoechoic cores corresponding to unruptured luteinized follicles and corpora lutea, respectively. On day 4 post-hCG, there was a substantial reduction in the number of antral follicles. In stimulated animals, follicular growth, ovulation, and formation of luteal structures were accompanied by corresponding physiological changes in the serum estradiol and progesterone profiles. These findings, for the first time, showed that ultrasonographic imaging approach is useful for precise monitoring of temporal changes in follicular developmental dynamics and to time the hCG induced ovulation in the bonnet monkey.

Andrology laboratory review: Evaluation of sperm concentration.

This article is the result of the work of the andrology task-force of the Association of Applied Animal Andrology, American College of Theriogenologists, European College of Animal Reproduction, Society for Theriogenology, and National Association of Animal Breeders. It is intended to serve as a comprehensive reference on methods to evaluate sperm concentration and to contribute to the adoption of best practices in veterinary andrology laboratories. The information covered in the article includes sample preparation and the use of manual counts, spectrophotometers, computer-assisted semen analysis, NucleoCounter, and flow cytometry. Emphasis is given to the principles of the methods and equipment, performing the evaluation, and common mistakes and/or pitfalls. In addition, the precision and accuracy of the different methods are also discussed.

Astaxanthin present in the maturation medium reduces negative effects of heat shock on the developmental competence of porcine oocytes.

Astaxanthin, one of the most common carotenoids, elicits antioxidant effects on cellular viability and embryonic development. This study was conducted to investigate the effects of astaxanthin on maturation, fertilization and development of porcine oocytes matured in vitro under heat stress conditions, and then fertilized and cultured under standard conditions. Porcine oocytes were cultured in maturation medium supplemented with different concentrations of astaxanthin (0, 0.25, 0.5 or 1 ppm) for 46 h at either 38.5 or 41 °C. In comparison to oocytes cultured at 38.5 °C, the exposure of porcine oocytes to 41.0 °C during in vitro maturation (IVM) significantly inhibited maturation and development of fertilized oocytes to the blastocyst stage. Supplementation of maturation medium with astaxanthin (0.5 ppm) significantly improved oocyte maturation, fertilization and development to the blastocysts stage in both oocyte groups. However, the total cell number and apoptosis index of blastocysts did not differ among groups. Moreover, astaxanthin (0.5 ppm) significantly increased the rate of oocytes that reached metaphase II and decreased proportion of apoptotic oocytes exposed to H2O2 (1.0mM) during IVM. In summary, we demonstrated that supplementation of maturation medium with astaxanthin (0.5 ppm) exerted antioxidative effects and improved the ability of maturation, fertilization, and development of porcine oocytes exposed to heat stress.

Relaxin: a hormonal aid to diagnose pregnancy status in wild mammalian species.

In the beginning of 1960s, seminal studies characterizing circulating concentrations of immunoreactive relaxin in companion dogs and evaluating the differences in concentrations among pregnant, nonpregnant, and pseudopregnant bitches indicated the potential for relaxin to be applied clinically as a diagnostic aid to detect pregnancy status in wild animal species. A brief historical overview of the nature of relaxin and early work to develop and validate immunologic methods to analyze relaxin in the blood of rodents and pigs is initially discussed, which is followed by a summary of the development and validation of relaxin immunoassays to diagnose pregnancy in companion dogs and cats. Thereafter, observation of the pregnancy-specific increase in circulating concentrations of relaxin in laboratory, companion, and farm animal species leads to discussion on the application of radioimmunoassays, enzyme immunoassays, and a rapid immunomigration assay to diagnose pregnancy in wild terrestrial (e.g., wolves, lions, elephants, rhinoceros, panda) and marine (e.g., seals, dolphins) mammal species. A reference table is included with a comprehensive list of numerous species and essential reagents that have been used in various in-house and commercial immunoassays to successfully analyze relaxin quantitatively and qualitatively in blood (serum or plasma) and to some extent in urine. Although the detection of relaxin concentrations has the potential to aid in the diagnosis of pregnancy in many wild animal species, there are challenges in other species. Future efforts should focus on validation of nonradiolabeled relaxin immunoassays for broader application among species and improving techniques (e.g., extraction, purification) to analyze relaxin in samples other than blood (e.g., urine, feces, saliva, blow, skin, blubber) that can be collected in a less-invasive or -stressful manner and processed accordingly for basic and applied purposes, especially with application toward conservation of threatened or endangered species.

Bovine placenta: a review on morphology, components, and defects from terminology and clinical perspectives.

The bovine placenta has been the subject of many studies. Concurrently, several specialized terms have been developed to describe its development, morphology, components, function, and pathology. Many of these terms are simple, some are difficult to understand and use, and others are antiquated and may not be scientifically accurate. Defining and adopting terminology for the bovine placenta that is clear, precise and understandable, and available in a single source is expected to facilitate exchange of clinical and research information. This review presents a brief overview of the current knowledge regarding the bovine placenta and attempts to define terms. In this process, conventional terminology is presented, and contemporary and novel terms are proposed from a biological perspective. For example, use of terms such as syndesmochorial, retained placenta, and large offspring syndrome should be revisited. Furthermore, the clinical relevance of the structure and function of the bovine placenta is reviewed. Finally, terms discussed in this review are summarized (in table format).

Cyclic estrous-like behavior in a spayed cat associated with excessive sex-hormone production by an adrenocortical carcinoma.

A 15-year-old, spayed female domestic shorthair cat was evaluated for 1-year duration of cyclic intermittent estrous behavior. Diagnostic testing performed before referral, including baseline progesterone concentration, human chorionic gonadotropin (hCG) hormone stimulation test and surgical exploratory laparotomy, had remained inconclusive for a remnant ovary. Evaluation of sex hormones before and after adrenocorticotropic hormone (ACTH) administration revealed increased basal concentrations of androstenedione, estradiol, progesterone, and 17α-hydroxyprogesterone and normal ACTH-stimulated hormone concentrations. Enlargement of the right adrenal gland was identified by abdominal ultrasound. The cat underwent an adrenalectomy and histopathology of the excised adrenal gland was consistent with an adrenocortical carcinoma. Clinical signs resolved immediately following surgery, and most hormone concentrations declined to within or below the reference interval (RI) by 2 months after surgery.

Catheter related bacteremia in rats: A preliminary report on the effect of methylene blue coated catheter.

The safety and efficacy of methylene blue (MB) coated indwelling jugular vein/cranial vena cava catheter made up of polyurethane material was tested in a rat model, receiving bacterial culture suspension of Pseudomonas aeruginosa, and Staphylococcus aureus. Daily blood samples were collected from the catheter and peripheral vein for bacterial culture. The clinical parameters (rectal temperature, respiratory rate, total white blood cell count, and loss in body weight) were not different between the groups. All the rats became bacteremic with similar changes in the number of colony forming units in the catheter and peripheral samples. Histopathological lesions were not different between the groups. The findings suggest that rats receiving MB coated catheters behaved similar to non-coated catheters. Based on the results it can be concluded that for this type of gross contamination, catheter coating alone may not eliminate infection/bacteremia.

Effect of caspase inhibitors on the post-thaw motility, and integrity of acrosome and plasma membrane of cryopreserved equine spermatozoa.

The present study was designed to test the hypothesis that addition of anticaspase cocktails (inhibiting caspases and thus blocking apoptosis) to the extenders increases the post-thaw viability of equine spermatozoa. The addition of caspase inhibitors failed to improve the acrosome and plasma membrane integrity of spermatozoa, suggesting that in equine sperm cryopreservation protocols, the addition of these caspase inhibitors to cryopreservation medium may not be beneficial in protecting the sperm from the stress of cryopreservation.

Efficacy of the anticaspase agent zVAD-fmk on post-thaw viability of canine spermatozoa.

Cryopreservation protocols for gametes are constantly improved with the aim of increasing the post-thaw viability of gametes. It is becoming clear that stress, resulting from cryopreservation, reduces cell numbers by apoptosis. Apoptosis, or programmed cell death, is a gene-activated event that occurs as a normal consequence of development and as a result of cellular stress. Apoptosis is mediated by the family of cysteine-dependent asparate-specific proteases (caspases). The present study was designed to test the hypothesis that addition of an anticaspase (zVAD-fmk) that has inhibitory properties against caspases and apoptosis to semen extenders and to the thaw medium would increase post-thaw viability of canine spermatozoa. Extenders were added in a two-step process. A dose of 100 microM caspase inhibitor was used. Four groups (n=6 for each) were composed based on the presence or absence of the caspase inhibitor: Group I (control), no caspase inhibitor in the extender or the thaw medium; Group II, caspase inhibitor in the thaw medium; Group III, caspase inhibitor in Extender II; and Group IV, caspase inhibitor in both Extender II and the thaw medium. Post-thaw motility, plasma membrane integrity, and acrosome status were investigated. The addition of caspase inhibitor to Extender II or to the thaw medium failed to improve the parameters that were studied. The results suggest that this caspase inhibitor may not be beneficial to the post-thaw motility of canine spermatozoa if used at this concentration.