The Activity and Localization of 3β-hydroxysteroid Dehydrogenase/Δ5-Δ4 Isomerase and Release of Androstenedione and Progesterone by Uterine Tissues During Early Pregnancy and the Estrous Cycle in Pigs

Steroid hormones are produced by the porcine uterus. We hypothesized that the uterus in pigs possesses active 3β-hydroxysteroid dehydrogenase/Δ⁵-Δ⁴ isomerase (3β-HSD) responsible for progesterone and androstenedione production, that uterine steroids may supplement the amount of steroid hormones produced by embryos and corpus luteum and that these steroids are necessary for maintenance of pregnancy. In this study, we examined 1) endometrial and myometrial expression of 3β-HSD mRNA, 2) uterine 3β-HSD protein activity and 3) in vitro production of A₄ and P₄ by uterine slices harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle. The expression of 3β-HSD and the presence and activity of 3β-HSD protein were different in the endometrium and the myometrium during the examined periods of pregnancy and the estrous cycle. Production of A₄ by the endometrium and myometrium was highest on days 12 to 13 of pregnancy and the estrous cycle. Endometrial secretion of P₄ did not differ in the course of early pregnancy and on the respective days of the estrous cycle. The gravid myometrium was the highest source of P₄ in pregnant pigs on days 12 to 13. The release of P₄ by the cyclic myometrium rose during the examined days of the estrous cycle. The steroidogenic activity of the uterus, as described in this study, may support early pregnancy or the luteal phase of the estrous cycle in pigs.     

Pathological findings and probable causes of the death of Stejneger’s beaked whales (Mesoplodon stejnegeri) stranded in Japan from 1999 and 2011

One hundred and twenty stranding events of Stejneger’s beaked whales were reported in Japan between 1999 and 2011. The purpose of this study is to introduce pathological data and to discuss probable causes of death for 44 Stejneger’s beaked whales among them. The significant pathological findings were the pulmonary edema, parasitic granulomatous nephritis, emaciation, amyloidosis, suppurative bronchopneumonia and so on. The probable causes of death were categorized as noninfectious in 43 of the cases, which included drowning, starvation and secondary amyloidosis. One individual was diagnosed with septicemia, which was the only example of an infectious disease. Because we could not always perform advanced analyses, such as microbiology tests, biotoxin examinations or contaminant analyses, the finality of our findings may be impaired. However, the present study has broad implications on the causes of death of Stejneger’s beaked whales of the seas around Japan, which are valuable for the future studies and for the detection of emerging diseases.     

The Effect of Single Nucleotide Polymorphisms in the Tumor Necrosis Factor-α Gene on Reproductive Performance and Immune Function in Dairy Cattle

The present study aimed to assess the effect of polymorphisms in the tumor necrosis factor α (TNF-α) promoter (A/A, A/G and G/G) and exons (T/T, T/C and C/C) on immune function and reproductive performance in dairy cows. The occurrence of the first postpartum ovulation within 3 weeks in the cows with the TNF-α promoter A/G and G/G genotypes was higher than in the A/A group. Among the different TNF-α exon genotypes, the occurrence of early first postpartum ovulation was higher in the T/C and C/C genotype groups than in the T/T group. Single nucleotide polymorphisms (SNPs) in the TNF-α gene did not affect the rate of artificial insemination (AI) or duration from parturition to next conception (days open). The apoptosis rate of polymorphonuclear leukocytes (PMNs) did not differ among the TNF-α promoter genotypes, but the PMN transmigration rate was significantly higher for the A/A and A/G genotypes than for the G/G genotype. Interleukin 8 (IL-8) mRNA expression in PMNs and peripheral blood mononuclear cells (PBMCs) before culture was significantly higher for the A/A genotype compared with the G/G genotype. There were no significant differences between the genotypes in the mRNA expression of TNF-α, IL-6, IL-1β, and toll-like receptor 4 (TLR4) in PMNs and PBMCs before and 4 h after culture. IL-8 and IL-1β production by PBMCs cultured for 4 h was significantly higher for the animals with the A/A genotype than for those with the G/G genotype. On the other hand, no significant difference was observed in IL-8 and IL-1β production by PMNs among different TNF-α genotypes. Taken together, these results suggest that SNP in the TNF-α gene affects immune function and reproductive performance in dairy cows.     

Relationships Between the Appearances and Changes of Estrous Signs and the Estradiol-17β Peak,Luteinizing Hormone Surge and Ovulation During the Periovulatory Period in Lactating Dairy Cows Kept in Tie-stalls

Lactating Holstein-Friesian cows kept in tie-stall barn were used as subjects in this study. Rectal examination, ultrasonography and blood sampling were conducted every other day and then daily after the day on which diameter of the corpus luteum decreased. After the luteal diameter decreased for 2 consecutive days, rectal and ultrasound examinations, blood sampling, and observation of estrous signs were conducted at 6-h intervals. Most of the estrous signs became obvious with the increase in estradiol-17β (E₂) and became most remarkable 24 to 30 hours before ovulation, at which point the E₂ peak and luteinizing hormone (LH) surge were achieved, and then weakened which progression to ovulation. The correlation between the intensity of four estrous signs (hyperemia and swelling of the intravaginal part of the uterus, opening of the external uterine orifice and viscosity of the cervical mucus) and the plasma E₂ concentration was higher than that of three estrous signs (swelling of the vulva, contraction of the uterus, diameter of uterine horn) and the plasma E₂ concentration. The relaxation of the intravaginal part of the uterus showed a unique change compared with the other estrous signs, and it became most obvious 6, 12 and 18 h before ovulation; this obviously relaxed period was consistent with the generally accepted theoretical optimal time for artificial insemination (AI), i.e., 6 to 24 h after initiation of estrus. These results suggest that observation of estrous signs by vaginoscopic examination gave useful information for detection of the optimal timing of AI in the periovulatory period in lactating dairy cows kept in a tie-stall barn.     

Effect of Conceptus on Transforming Growth Factor (TGF) β1 mRNA Expression and Protein Concentration in the Porcine Endometrium—In Vivo and In Vitro Studies

Transforming growth factor (TGF) β and its receptors are expressed at the conceptus-maternal interface during early pregnancy in the pig. The present studies were conducted to examine: (1) the effect of conceptus products on TGFβ1 mRNA expression and protein concentration in the porcine endometrium using in vivo and in vitro models, and (2) the effect of TGFβ1 on proliferation of porcine trophoblast cells in vitro. During in vivo experiments, gilts with one surgically detached uterine horn were slaughtered on days 11 or 14 of the estrous cycle and pregnancy. For in vitro studies, endometrial explants and luminal epithelial (LE) cells co-cultured with stromal (ST) cells were treated with conceptus-exposed medium (CEM). Moreover, porcine trophoblast cells were treated with TGFβ1, and the number of viable cells was measured. On day 11, the presence of conceptuses had no effect on TGFβ1 mRNA expression, but decreased the TGFβ1 protein concentration in the connected uterine horn compared with the detached uterine horn. In contrast to day 11, on day 14 after estrus, TGFβ1 mRNA expression and protein content in the endometrium collected from the gravid uterine horn were greater when compared with the contralateral uterine horn. The treatment of endometrial slices with CEM resulted in greater TGFβ1 mRNA expression and protein secretion. LE cells responded to CEM with an increased TGFβ1 mRNA level. Moreover, TGFβ1 stimulated the proliferation of day 14 trophoblast cells. In summary, porcine conceptuses may regulate TGFβ1 synthesis in the endometrium at the time of implantation. TGFβ1, in turn, may promote conceptus development by increasing the proliferation of trophoblast cells.     

Intrafollicular Concentrations of Steroid Hormones and PGF2α in Relation to Follicular Development in the Mares during the Breeding Season

The concentrations of androstenedione, estradiol-17β, progesterone and PGF_2α contained in the follicular fluid produced by the follicles in collected ovaries of mares that have had estrous phase during the breeding season were measured and analyzed the relation between the growth stage of follicles and the hormone levels in the follicular fluid. An ultrasonographic diagnostic instrument was used to measure the diameter of the follicles in order to categorize the follicles into three groups the following: 8 small follicles (from 1.0 to less than1.5 cm), 8 medium follicles (from 1.5 to less than 3.0 cm), and 8 large follicles (from 3.0 to 5.0 cm), respectively. The analysis of the follicular fluid in ovaries of estrous mares showed that the concentrations of androstenedione were significantly higher in the medium or large follicles than in the small follicles and the concentrations of estradiol-17β were significantly higher in larger follicles than in the small or medium follicles (P<0.05). The concentrations of progesterone and PGF_2α, on the other hand, did not significantly vary regardless of follicluar size. In the follicles within the mare ovaries that have had estrous stage, the concentrations of the hormones related the ovulation, namely androstenedione and estradiol-17β, were higher with larger follicles.     

Number of ovulations in culled Landrace × Yorkshire gilts in the tropics associated with age,body weight and growth rate

The objective of the present study was to determine the number of ovulations in culledLandrace × Yorkshire (LY) crossbred gilts in the tropics associated with age, body weightand growth rate. The genital organs from 316 gilts were examined for gross abnormalities,and those with normal cyclic ovaries (n=155, 307 ± 4.1 days of age, 148 ± 1.6 kg bodyweight) were included in the analyses. Number of ovulations was defined by a count of thecorpora lutea (CL) from both ovaries. On average, the number of ovulations in LY gilts was15.9 ± 0.3 (range 4 to 27). The number of ovulations correlated with the body weight(r=0.31, P<0.001) and growth rate (r=0.20, P=0.015)of the gilts, but not with their age (P>0.05). Gilts with a bodyweight of 141 to 150 kg (17.0 CL, n=31) ovulated more than those with a body weight ≤130kg (14.1 CL, P=0.014, n=23). In conclusion, both the body weight andgrowth rate of the gilts were significantly correlated with the number of ovulations. Themaximum number of ovulations was found in gilts at a body weight of above 141 kg.     

Reproduction and Development of the Released Przewalski’s Horses (Equus przewalskii) in Xinjiang,China

In China, the first Przewalski’s horse (Equus przewalskii) group was released in Kalamaili Ungulate Protected Area in Xinjiang, in August 2001. The objective of this study was to investigate reproduction and development of released Przewalski’s horses in Xinjiang, China from 2002 to 2006. Twenty-four descendants were naturally born, average reproduction rate was 38.7%, and average survival rate of foals was 69.1% in this interim. Frequent alternation of the leading stallion and infertility in female horses due to environmental factors were main causes for the low reproduction rate. The infant mortality rate of released Przewalski’s horses was 25.0%, and 83.3% of death in infants was due to the leading stallion infanticide. The released Przewalski’s horses exhibited seasonal breeding, 70.8% of infants were born in May and June. The fertility rates were 8.3% in April and 37.5% in May, which were lower compared to those of the captive Przewalski’s horse groups (18.3% and 44.3%, respectively). Furthermore, the fertility rate in June was 33.3% and higher than the captive groups (18.3%). These findings showed that the breeding peak of the released Przewalski’s horse groups was later than the captive groups, and suggested that the altered survival environment and food supplies were the main reasons contributing to the delayed breeding peak.     

Expression of Glucocorticoid Receptor α and Its Regulation in the Bovine Endometrium: Possible Role in Cyclic Prostaglandin F2α Production

Cortisol (Cr), the most important glucocorticoid (GC), is well known to suppress uterine prostaglandin F2α (PGF) production. However, the details of the regulatory mechanisms controlling the cyclic changes in endometrial PGF production remain unclear. Here we investigated the expression of the GC receptor (GC-Rα), the actions of cortisol throughout the estrous cycle and the regulatory mechanism of GC-Rα in the bovine endometrium. The levels of GC-Rα protein were greater at the mid-luteal stage (Days 8–12) than at the other stages. Cr more strongly suppressed PGF production at the mid-luteal stage than at the follicular stage. GC-Rα expression was increased by progesterone (P4) but decreased by estradiol-17β (E2) in cultured endometrial stromal cells. The overall results suggest that ovarian steroid hormones control the cyclic changes in endometrial PGF production by regulating GC-Rα expression in bovine endometrial stromal cells.     

Genetic structure of the Amur tiger (Panthera tigris altaica) population: Are tigers in Sikhote‐Alin and southwest Primorye truly isolated?

We used molecular genetic analyses to noninvasively identify individual Amur tigers and define subpopulations of tigers in the Russian Far East. We identified 63 individuals after genotyping 256 feces, 7 hair and 11 blood samples collected within southern, central and northern Sikhote‐Alin, as well as Southwest Primorye. Analysis of nuclear DNA at 9 microsatellite loci demonstrated greater genetic similarity between animals from southern and northern Sikhote‐Alin (some 500 km apart) than between animals from Ussuriskii State Nature Reserve and Southwest Primorye (less than 10 km apart at their nearest point), suggesting that a true barrier exists preventing movements of tigers between Southwest Primorye and the southern Sikhote‐Alin Mountains.     

β-carotene and retinol contents in the meat of herbivorous ungulates with a special reference to their public health importance

The aim of this study was to estimate total carotenoids, β-carotene and retinol concentrations in the liversand muscles of some ungulates (cattle, buffalo, sheep, goats and horses) in comparison with the Wistar rats asa control. Cattle and horses had the highest contents of total carotenoids and β-carotene. Unexpectedly, sheepwas the highest accumulator of retinol with a mean concentration of 203 ± 23.34 µg/g, whilethe least accumulator was buffalo, having a mean value of 58.28 ± 13.77 µg/g. Livers hadhigher contents of the examined phytochemicals than muscles. Consumption of these tissues may provide humanwith some needs from these important phytochemicals, though ingestion of livers, especially that of the sheep,is not advisable for the pregnant women.     

Comparison of a 3-hole 4.5-mm Dynamic Compression Plate and a 7-hole 5.5-mm Y Locking Compression Plate for Arthrodesis of the Proximal Interphalangeal Joint in Horses—an Ex Vivo Biomechanical Study

The objective of this study was to compare the biomechanical properties in a single cycle axial loading test and the types of failures in two constructs (a 3-hole 4.5-mm dynamic compression plate (DCP) and 7-hole 5.5-mm Y locking compression plate (Y-LCP)) in equine proximal interphalangeal joint (PIJ) arthrodesis. One limb in each pair was randomly assigned to PIJ arthrodesis using a 3-hole 4.5-mm DCP combined with two transarticular 5.5-mm cortical screws, whereas the contralateral limb was submitted to PIJ arthrodesis using a 7-hole Y-shaped 5.0-mm LCP in conjunction with one transarticular 4.5-mm cortical screw inserted through the central plate hole. Cortical screws were inserted in lag fashion. Constructs were submitted to a single axial load cycle to failure. Construct stiffness, load, and deformation were analyzed. Dynamic compression plate and Y-LCP arthrodesis constructs did not differ significantly and were equally resistant to axial loading under the conditions studied (DCP and Y-LCP group stiffness, 5685.22 N/mm and 6591.10 N/mm, respectively). Arthrodesis of the PIJ using a DCP and two transarticular 5.5-mm cortical screws or a Y-LCP yielded biomechanically equivalent outcomes under the test conditions considered. However, Y-LCP provides less impact in the palmar/plantar bone. Application of Y-LCP with unicortical screws has equivalent biomechanical characteristics of DCP and may be a safe option for PIJ arthrodesis, where potential trauma secondary to applying bicortical screws in the palmar/plantar aspect of the pastern can be avoided.