A.T. Peter

Postpartum anestrus in dairy cattle

Fertility of the postpartum period is negatively influenced by the incidence of anestrus. The latter condition is characterized by the absence of estrous behavior, which may be an indication of suboptimal conditions (e.g., inadequate peripartum nutrition) or pathologic conditions (e.g., chronic debilitating diseases or uterine and ovarian diseases). Although initiation of ovarian follicular growth in the postpartum period is generally not affected, subsequent development (deviation) and the fate of the dominant follicle are the primary factors that affect reestablishment of ovarian cyclicity. Anestrus can be classified based on the three functional states of follicular development; that is, follicle emergence, deviation, and ovulation. Prevention of anestrus is preferable to treatment and can be achieved in part by maintaining a healthy periparturient period. To better understand the etiology of anestrus and its prevention, research is urgently needed in the following three areas: the role of peripartum disease conditions that influence reproduction, genes involved in ovulation, and the influence of proteins (e.g., leptin) that appear to be important links between metabolic signals and the neuroendocrine axis.

Effect of developmental stage on bovine oocyte plasma membrane water and cryoprotectant permeability characteristics

Knowledge of bovine oocyte plasma membrane permeability characteristics at different developmental stages in the presence of cryoprotective agents (CPAs) is limited. The objective of this study was to determine the oolema hydraulic conductivity (Lp), cryoprotectant permeability (PCPA), and reflection coefficient (σ) for immature (germinal vesicle stage, GV) and in vitro–matured (metaphase II, MII) bovine oocytes. Two commonly used cryoprotective agents, dimethyl sulfoxide (DMSO) and ethylene glycol (EG), were studied. Osmometric studies were performed using a micromanipulator connected to an inverted microscope at 22 ± 2°C. Each oocyte was immobilized via a holding pipette, and osmotically induced volume changes over time (dv/dt) were recorded. The Lp values for GV and MII oocytes in DMSO (LpDMSO) were 0.70 ± 0.06 and 1.14 ± 0.07 μm/min/atm (mean ± SEM) and in EG (LpEG) were 0.50 ± 0.06 and 0.83 ± 0.07 μm/min/atm, respectively. Estimates of PDMSO for GV and MII oocytes were 0.36 ± 0.03 and 0.48 ± 0.03 μm/sec, and PEG values for GV and MII oocytes were 0.22 ± 0.03, 0.37 ± 0.03 μm/sec, respectively. The σ values for GV and MII oocytes in DMSO (σDMSO) were 0.86 ± 0.03 and 0.90 ± 0.04 and in EG (σEG) were 0.94 ± 0.03 and 0.76 ± 0.04, respectively. These data demonstrate that bovine oolema permeability coefficients to water and cryoprotectants change after in vitro maturation. Furthermore, the bovine oocyte PDMSO is higher than the PEG. These results may provide a biophysical basis for developing criteria for choosing optimal CPAs and for minimizing damage during addition and removal of the CPAs. Additionally, these data support the hypothesis that different procedures may be required for optimal cryopreservation of different oocyte developmental stages. Mol. Reprod. Dev. 49:408–415, 1998.

Compilation of classical and contemporary terminology used to describe morphological aspects of ovarian dynamics in cattle

Veterinarians and scientists involved in applied and basic research in cattle require a lexicon of terms that is used uniformly so that diagnoses and inference of results between and among studies can be correctly interpreted and substantiated or negated and therapy and hypotheses can be formulated without unnecessary confusion and redundancy in treatments and experiments. This review provides a compilation of many of the classical and contemporary terms used in association with ovarian dynamics primarily during the estrous cycle in cattle, which can also apply to other reproductive states. While many classical terms used to describe healthy and diseased conditions associated with follicles and corpora lutea are still applicable today, there are some that have become antiquated (e.g., cystic corpus luteum, cystic ovarian degeneration, luteolysis, and granulosa cell tumor), due, in part, to advanced technology (e.g., ultrasonography) and a more thorough understanding of ovarian function. In this regard, older terms have been revised (e.g., corpus luteum with a cavity, follicular and luteinized-follicular cysts, structural and functional luteal regression, and granulosa-theca cell tumor) and newer terms have been coined (e.g., follicle deviation) and advocated herein. Defining and adopting terminology used in bovine reproduction that is clear, precise and understandable and available in a single source, is expected to make the exchange of clinical and research information and outcomes more effective, safe, and economical.

Absorption of Escherichia coli endotoxin (lipopolysaccharide) from the uteri of postpartum dairy cows.

An experiment was conducted to test the hypothesis that Escherichia coli (E. coli ) endotoxin is readily absorbed from uteri of early postpartum cows and that the absorbed endotoxin provokes systemic relcase of prostaglandins. Eleven postpartum Holstein dairy cows (aged 3 to 7 yr) with normal puerperium were selected and divided into a treatment group (n=7), which received intrauterine infusions of E. coli endotoxin, and a control group (n=4), which received intrauterine infusions of 10 ml of saline on Days 5 and 20 post partum. Blood samples were collected once every 30 min for 6 h starting from the time of infusion. Harvested sera samples were analyzed for concentrations of stable metabolites of prostacyclin (PCM), prostaglandin F(2alpha) (PGFM), and thromboxane A(2) (TXB(2)). Plasma samples were qualitatively tested for the presence of endotoxin. Endotoxin was detected in the plasma samples of cows that received endotoxin on Day 5 post partum 4 h after the infusion. Endotoxin was not detected in any of the samples from control cows on Days 5 and 20 post partum or from treatment group cows on Day 20 post partum. Cows treated on Day 5 post partum showed increases in serum PGFM concentrations from 710 +/-64pg/ml to peak concentrations of 1223 +/- 47 pg/ml within 2 h, followed by a decline to baseline concentrations within 4 h. The amount of PGFM released in treated cows on Day 5 post partum was higher (P < 0.05) than in control cows on Day 5 or in treated and control cows on Day 20 post partum. Serum PCM concentrations increased from 156+/-24 pg/ml to peak concentrations of 1348+/-127 pg/ml within 1 h. The amount of PCM released in treated cows on Day 5 postpartum was higher (P< 0.05) than in control cows on Day 5 or in treated and control cows on Day 20 post partum. The TXB(2) concentrations increased from 315+/-38 pg/ml to peak concentrations of 5043 +/- 242 pg/ml within 1 h and fell to baseline concentrations within 5 h. The amount of TXB(2) concentrations released in treated cows on Day 5 post partum was significant (P < 0.05) compared with those of cows in the other groups. The results support the hypothesis that uteri of early postpartum cows are capable of absorbing endotoxin, and the absorbed endotoxin provokes changes in the serum concentrations of prostanoids.

Apoptosis of ovarian granulosa cells: correlation with the reduced activity of ERK-signaling module.

Apoptosis of the ovarian granulosa cells plays a crucial role in the determination of the number of follicles destined to ovulate in each reproductive cycle. While the activation of specific apoptotic pathway or the inactivation of cell survival pathway can initiate apoptosis, the signaling mechanism(s) involved in initiating the onset of apoptosis in granulosa cells is not fully understood. In the present study, using granulosa cells derived from eCG‐primed immature rats, we investigated the temporal signaling events involved in the onset of apoptosis in the granulosa cells. The administration of 15 IU of eCG to 21‐day‐old immature female rats stimulate the growth and development of ovarian follicles until 72 h, after which the granulosa cells of the ovarian follicles undergo apoptosis due to the waning levels of tropic hormonal support. An analysis of the signaling events leading to apoptosis indicates that the DNA fragmentation can be seen in these cells from 96 h. A small increase in the levels of the pro‐apoptotic factor Bax can be seen from 96 h while an increase in the activity of JNK can be seen from 108 h onwards. By contrast, a reduction in ERK signaling can be seen by 48 h. Similar reduction in Raf‐1 kinase activity can be discerned from 48 h onwards. A concomitant decrease in the phosphorylated form of Bad can also be detected. These findings taken together, suggest that the loss of tropic hormone support is translated into the attenuation of Raf‐1‐MEK‐ERK signaling pathway and this reduction along with a reduction in the levels of phosphorylated form of Bad triggers the onset of apoptosis in the ovarian granulosa cells. J. Cell. Biochem. 75:547–554, 1999.

REPRODUCTION IN LLAMAS AND ALPACAS : A REVIEW

In this review we attempt to compile and summarize the diverse and often contradictory material presented on the reproduction of llamas and alpacas (hereafter referred to as lamoids). Lamoids have recently gained international popularity, and theriogenologists are often asked to intervene in clinical management of reproductive problems of these animals. We therefore present a discussion of the reproductive anatomy, physiology, and behavior of llamas as well as the follicular dynamics as observed with ultrasonography. The nonsurgical embryo transfer procedure and the nutrient requirements of llamas are also discussed.

Fractionation of bovine spermatozoa for sex selection: A rapid immunomagnetic technique to remove spermatozoa that contain the H-Y antigen

Abstract A study was conducted to rapidly fractionate bovine spermatozoa on the basis of cell-surface H-Y antigen (i.e., Y chromosome-bearing spermatozoa). A novel, rapid immunomagnetic method was developed for removal of spermatozoa that bound to anti-H-Y IgG. Fluorescent labeling and flow cytometry were used to measure the efficiency with which spermatozoa binding to anti-H-Y were removed by the immunomagnetic technique. Washed bovine spermatozoa (n=7 bulls) were treated with a mouse monoclonal IgG antibody to H-Y antigen (MoAb 12 49 ). Fluorescent labeled goat antibody against mouse IgG was added to label those spermatozoa with cell-surface H-Y antigens. Supermagnetized polymer beads coated with an anti-antibody to the MoAb 12 49 were then added to the spermatozoa. After 20 min of incubation, spermatozoa were exposed for 2 min to a magnet, causing the magnetized particles to adhere to the sides of the tube. Nonmagnetized spermatozoa in the supernatent were aspirated and analyzed for fluorescent label by flow cytometry. Approximately 50% of spermatozoa not subjected to immunomagnetic separation were fluorescent labeled, and about one-half of the spermatozoa were observed microscopically to be bound to the magnetized polymer beads prior to magnetic separation (P

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.

Cryobiology of Rat Embryos I: Determination of Zygote Membrane Permeability Coefficients for Water and Cryoprotectants, Their Activation Energies, and the Development of Improved Cryopreservation Methods

Abstract New rat models are being developed at an exponential rate, making improved methods to cryopreserve rat embryos extremely important. However, cryopreservation of rat embryos has proven to be difficult and expensive. In this study, a series of experiments was performed to characterize the fundamental cryobiology of rat fertilized 1-cell embryos (zygotes) and to investigate the effects of different cryoprotective agents (CPAs) and two different plunging temperatures (Tp) on post-thaw survival of embryos from three genetic backgrounds. In the initial experiments, information on the fundamental cryobiology of rat zygotes was determined, including 1) the hydraulic conductivity in the presence of CPAs (Lp), 2) the cryoprotectant permeability (PCPA), 3) the reflection coefficient (σ), and 4) the activation energies for these parameters. PCPA values were determined for the CPAs, ethylene glycol (EG), dimethyl sulfoxide (DMSO), and propylene glycol (PG). Using this information, a cryopreservation method was developed and the cryosurvival and fetal development of Sprague-Dawley zygotes cryopreserved in either EG, DMSO, or PG and plunged at either −30 or −80°C, were assessed. The highest fetal developmental rates were obtained using a Tp of −30°C and EG (61.2% ± 2.4%), which was not different (P > 0.05) from nonfrozen control zygotes (54.6% ± 3.0%).

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).