Relationship Between Fertility at Foal Heat and Blood Biochemistry Parameters Monitored During the Peripartum Period in Thoroughbred Mares

This study aimed to examine fertility at foal heat and its relevance to body condition score (BCS) and blood nutritional metabolites in Thoroughbred mares. Thoroughbred mares foaled from 2006 to 2009 were included and classified into two groups: group C (conception; n = 34), which included mares that conceived during foal heat (within 3 weeks after foaling), and group NC (nonconception; n = 39), which included mares that did not conceive despite mating during their foal heat. BCS and blood samples were obtained 1 month before the expected foaling date and 1, 2, 3, and 4 weeks after foaling. Total protein (TP), albumin (Alb), blood urea nitrogen (BUN), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), total cholesterol (T-Cho), triglyceride (TG), nonesterified fatty acid (NEFA), calcium (Ca), inorganic phosphate (iP), and magnesium (Mg) levels were measured using an automatic clinical chemistry analyzer. Repeated measures analysis of variance (ANOVA) and Student t-test were used to examine the differences between the two groups. No significant differences were observed between the two groups in any of the above-mentioned parameters at 1 month before the expected foaling date (Student t-test). Furthermore, no significant differences were observed between the two groups in serum levels of TP, Alb, AST, GGT, T-Cho, NEFA, Ca, and Mg and BCS at postpartum periods (repeated measures ANOVA). Serum TG, BUN, and iP levels remained lower in group NC than in group C after foaling (P < .05, repeated measures ANOVA). Although the mechanism by which these nutritional factors affect a decline in reproductive performance remains unclear, our results suggest that blood biochemical tests can detect potential imbalances in nutrition and metabolism, even if there is no difference in BCS.     

Fertilization Ability of Porcine Oocytes Reconstructed from Ooplasmic Fragments Produced and Characterized after Serial Centrifugations

Mitochondria are reported to be critical in in vitro maturation of oocytes and subsequent embryo development after fertilization, but their contribution for fertilization has not been investigated in detail. In the present study, we investigate the contribution of mitochondria to fertilization using reconstructed porcine oocytes by fusion of ooplasmic fragments produced by serial centrifugations (centri-fusion). Firstly, we evaluated the characteristics of ooplasmic fragments. Three types of fragments were obtained by centrifugation of porcine oocytes matured in vitro for 46 h: brownish (B), transparent (T) and large (L) fragments containing both B and T parts in a fragment. The production efficiencies of these types of fragments were 71.7, 91.0 and 17.8 fragments/100 oocytes, respectively. In experiments, L fragments were excluded because they contained both brownish and transparent components that were apparently intermediate between B and T fragments. Observations by confocal microscopy after staining with MitoTracker Red CMXRos^® and transmission electron microscopy revealed highly condensed active mitochondria in B fragments in contrast to T fragments that contained only sparse organelles. We reconstructed oocytes by fusion of a karyoplast and two cytoplasts from B and T fragments (B and T oocytes, respectively). The B oocytes showed higher sperm penetration (95.8%) and male pronuclear formation rates (94.2%) by in vitro fertilization than T oocytes (66.7% and 50.0%, respectively). These results suggest that the active mitochondria in oocytes may be related to their ability for fertilization.     

Modified resazurin microtiter assay for in vitro and in vivo assessment of sulfamonomethoxine activity against the fish pathogen <Emphasis Type="Italic">Nocardia seriolae</Emphasis>

Resazurin microtiter assay (REMA) was carried out using four sulfonamides, three culture media, and four inoculum sizes as a first screening step to establish an easy-to-interpret sulfonamides susceptibility testing method for Nocardia seriolae. The in vitro activity of sulfamonomethoxine (SMM) against 190 clinical N. seriolae isolates was then examined, and in vivo experimental treatment was performed. When the culture medium and the inoculum size were considered in tandem, a 0.5× the original concentration of cation-adjusted Mueller–Hinton broth and an inoculum size of 10² CFU/well showed the clearest endpoint reading for all tested drugs, and the REMA-generated data were in excellent agreement with those generated by the reference Etest method. SMM activity showed minimum inhibitory concentration (MIC) values of 4–32 μg/ml against all tested N. seriolae isolates. Treatment of amberjack groups experimentally infected with N. seriolae isolates having SMM MICs of 4 and 32 μg/ml, resulted in survival rates of 100% and 87.5% in the two groups, respectively. In this study, we developed a simple visual method to test SMM activity against N. seriolae.     

Evaluation of nutritional values of selected commercial fish meal sources in turbot (Psetta maxima) diets

This study was conducted to evaluate nutritional values of selected commercial fish meals (FM) with two experiments in diets of turbot. In experiment I, five diets based on two Peruvian anchovy meals (one is a flame‐dried FM: Peru‐1 and the other steam‐dried FM: Peru‐2), a Danish FM (Danish LT), a domestic anchovy LT meal (Domestic LT) and an Alaskan white FM (WFM) as major protein source were formulated to contain 56% protein. The diets were fed to fish with an initial average weight of 119.4 ± 0.1 g for 9 weeks. Growth rate, feed utilization, whole body and visceral compositions, nitrogen, lipid and energy balances did not significantly change in Experiment 1 among the treatments. In Experiment 2, four diets based on three FMs (Domestic LT, Danish LT and Peru‐1) and their mixture (Mix FM) were formulated to contain 50% protein and fed to triplicated groups of fish with an average initial weight of 51.4 ± 0.1 g for 6 weeks. Once again, there were no significant differences in growth rate and feed consumption among the treatments. However, fish on Mix FM diet had significantly better FCR and PER than the others. The results of two experiment indicated that Danish LT and Domestic LT were better in terms of phosphorus (P) retention and losses than two Peruvian FMs and WFM, suggesting that although different sources of FMs were comparable in terms of growth and feed utilization, they could lead to substantial differences in environmental impacts.     

Genetic characterization of the endangered Kiso horse using 31 microsatellite DNAs

In order to contribute to conservation of the endangered Kiso horse, we clarified their genetic information using 31 microsatellite DNAs, and genotyped 125 horses, 83% of the existing breed. First, we clarified the current status of the horses. The horses were confirmed to have experienced rapid loss of population causing a bottleneck, and their effective population size was much smaller than their census size. Moreover, the number of alleles (6.3), observed heterozygosity (0.674), and expected heterozygosity (0.662) were in the same range as other endangered horses all over the world. Therefore, although their inbreeding level was not so severe (F(is): -0.017), the Kiso horse is surely one of the endangered. Second, we obtained genetic information of individuals. This information allowed us to understand the genetic distance of individuals, and might help in development of a reproductive strategy concerning the genetic distance between the mating pairs. Moreover, there appeared to be 4 subpopulations of Kiso horse, and this result was in good agreement with their historical background. Third, we confirmed that the parentage test for identification using the 31 microsatellite DNAs was highly reliable (probability of exclusion: 0.999999993). This identification increases the reliability of stud certification, and is also helpful for effective management. Understanding the genetic diversity within the population and the relationships among individuals is important to ensuring effective management for maintenance of genetic variation, and this study may help in conservation of the endangered Kiso horse.     

Assessment for predicting parturition in mares based on prepartum temperature changes using a digital rectal thermometer and microchip transponder thermometry device

The purpose of this study was to observe the changes in body temperature before parturition using a wireless temperature monitoring device (WTMD) and to evaluate the usefulness of body temperature measurements using a digital rectal thermometer (DRT) and a microchip transponder thermometry device (MTTD) for predicting parturition in mares. The body temperatures using a WTMD at 0 hr and -1 hr were significantly different from those at the same time on Days 1-5 (P<0.01). The temperature differences between the morning of Day 0 and at -3 hr, -2 hr, -1 hr and 0 hr using the DRT and MTTD showed a significant drop compared with the temperature differences between the morning and evening of Days 1-5 (P<0.05). Furthermore, when the cutoff value of the temperature differences between the morning and other times was set to ≤0, the sensitivities of the DRT and MTTD in the evening of Day 0 and at -3 hr were 43% and 100% and 71% and 86%, respectively. The results suggested that monitoring the body temperature differences between morning and within 3 hr before the time of parturition is a valuable method for predicting parturition in mares. Conversely, this method would be more useful in predicting parturition when used in combination with other observations such as the mammary gland size and waxing of the teat ends because it has nearly a 20% probability of false-positive results prior to the day of parturition.     

Estimation of the internal shear strength distribution of the element for laminated veneer lumber by nonlinear least-squares method

Until now we developed an estimation method for strength distributions of laminated veneer lumber (LVL) element by nonlinear least-squares method (NLM). Estimated strengths by this method were modulus of elasticity (MOE) and modulus of rupture (MOR) in the horizontal use direction and the vertical use direction, tensile strength and compression strength. But to use LVL for structural members, shear strength was also needed. Therefore, we tried to estimate the shear strength distribution of LVL element by NLM same as MOE and MOR in the horizontal use direction and the vertical use direction, the tensile strength of LVL and the compression strength of LVL in the previous reports. We conducted shear strength test for LVL and estimated element shear strength distribution by LVL strength data in the horizontal and vertical use direction. Next, we simulated LVL shear strength distribution using element shear strength distribution and compared with experimental ones in each use direction. They were overlapped in both use direction. Therefore, we could validate NLM for estimating element shear strength distribution.