Evaluation of bovine embryos produced in high performance serum-free media

This review evaluates the quality of bovine embryos developed from in vitro-matured (IVM) and -fertilized (IVF) oocytes cultured in either serum-free or serum-containing media. Bovine embryos cultured in serum-supplemented medium contain numerous cytoplasmic lipid droplets and immature mitochondria compared to those cultured in serum-free medium. The accumulation of cytoplasmic lipids in embryos developed in serum-containing medium may be a result of incorporation of lipoproteins from the serum and may result in impaired function of mitochondria. The improved serum-free media (IVMD101 and IVD101) offer several advantages over culture in serum-containing medium, including increased rates of blastocyst formation and higher cell numbers. Additionally, the survival and hatching rates of embryos produced in serum-free media after post-thaw culture were superior to those of embryos produced in the serum-containing medium, suggesting that the abnormal accumulation of cytoplasmic lipids in embryos may have a negative effect on the sensitivity of embryos to chilling and freezing. These serum-free culture systems have proven to be beneficial for the production of good quality embryos from IVM-IVF bovine oocytes. Furthermore, recent studies have shown a correlation between mitochondrial function (oxygen consumption) and embryo quality. A new method using scanning electrochemical microscopy may be capable of assessing the viability and developmental potential of bovine embryos.     

Nationwide survey of hepatitis E virus infection among wildlife in Japan

In Japan, hepatitis E virus (HEV) causes hepatitis in humans through the consumption of raw or undercooked meat, including game meat. In the present study, nationwide surveillance of HEV infection among a total of 5,557 wild animals, including 15 species, was conducted in Japan. The prevalence of anti-HEV antibodies in wild boar was 12.4%, with higher positive rates in big boars (over 50 kg, 18.4%) than in small individuals (less than 30 kg, 5.3%). Furthermore, HEV RNA was more frequently detected in piglets than in older boars. Interestingly, the detection of HEV among wildlife by ELISA and RT-PCR suggested that HEV infection in Sika deer was a very rare event, and that there was no HEV infection among wild animals except for wild boar, Sika deer and Japanese monkeys. In conclusion, wild boar, especially piglets, are at high risk of HEV infection, while other wild animals showed less risk or no risk of HEV transmission.     

Correlation between villous height and the disaccharidase activity in the small intestine of piglets from nursing to growing

Early weaning induces villous atrophy in the small intestine. Reduction in villous height in the small intestine after weaning is associated with reductions in brush‐border enzyme activity. Body weight gain after weaning is, therefore, correlated with villous height. This evidence suggested that the maintenance of small intestinal structure and function after weaning is important for the growth of young pigs. On the other hand, the relationship between villous height and the activity of the digestive enzymes in the small intestine has not been studied with piglets from the suckling to the growing period. Five suckling piglets, four piglets in the proximal stage of weaning, four pigs in the distal stage of weaning and four growing pigs were used. The activities of lactase (LA), sucrase (SA) and maltase (MA) were determined. LA showed a positive correlation with villous height in weaning. SA and MA were positively correlated with villous height from suckling to growing. In a previous study, non‐infectious dyspeptic diarrhea was frequently observed in growing piglets on Japanese swine farms. The maintenance of villous height to retain disaccharidase activity may prevent dyspepsic diarrhea in this stage.     

Comparison of dendritic cell-mediated immune responses among canine malignant cells

Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials.     

Spatial and interspecific variability in phenological responses to warming temperatures

A comprehensive understanding of species phenological responses to global warming will require observations that are both long-term and spatially extensive. Here we present an analysis of the spring phenological response to climate variation of twelve taxa: six plants, three birds, a frog, and two insects. Phenology was monitored using standardized protocols at 176 meteorological stations in Japan and South Korea from 1953 to 2005, and in some cases even longer. We developed a hierarchical Bayesian model to examine the complex interactions of temperature, site effects, and latitude on phenology. Results show species-specific variation in the magnitude and even in the direction of their responses to increasing temperature, which also differ from site-to-site. At most sites the differences in phenology among species are forecast to become greater with warmer temperatures. Our results challenge the assertion that trends in one geographic region can be extrapolated to others, and emphasize the idiosyncratic nature of the species response to global warming. Field studies are needed to determine how these patterns of variation in species response to climate change affect species interactions and the ability to persist in a changing climate.     

Quantitative evaluation of the degree of sprout leaf bending of rice cultivars using P-type Fourier descriptors and principal component analysis

For decades, seedling peaches have been used as the standard rootstock in California almond orchards. Vigorous, deep rooted trees are needed in almond orchards for maximum yields and to withstand the annual tree-shaking at harvest. Currently, researchers are actively evaluating rootstocks for almonds in field trials and in various screening protocols. In this study, seedling rootstocks, obtained from male-sterile advanced generation peach-almond (PEAL) hybrid mother trees, were compared with ‘Nemaguard’ peach seedlings for emergence in the nursery row, trunk caliper at propagation time, and end of season dormant above ground tree weight. Seedling emergence was affected significantly (P ≤ 0.05) by seed source, as was trunk caliper and end of season dormant above ground tree weight. Trunk caliper and dormant above ground tree weight were also affected significantly (P ≤ 0.05) by planting year. Results obtained in this study demonstrate the enhanced first year growth from seedlings of advanced generation PEAL hybrids, as compared to ‘Nemaguard’ seedlings. Seedling emergence in the rootstock bed was not affected significantly by planting year. Advanced generation PEAL hybrid seedlings were ready for June-budding at an earlier date compared to ‘Nemaguard’ seedlings, providing the potential for larger-sized finished nursery stock by the end of the growing season. Due to the male-sterile status of the advanced generation PEAL mother trees, bloom periods of several root-knot nematode resistant rootstock cultivars were examined for their degree of synchronicity with the mother trees. The examined rootstocks and mother trees varied in both chill hour and post-chill heat requirements necessary to effect bloom. ‘Flordaguard’ peach rootstock began bloom in advance of the male-sterile mother trees, whereas the bloom period of ‘Tsukuba No. 4’ occurred well after, suggesting they would not be effective synchronous pollenizers for consistent hybrid seed production. Based on more limited flowering period data, better bloom synchronicity was achieved with a Tsukuba No. 4 X Flordaguard hybrid.     

Effect of atropine and pyridostigmine on growth hormone response to GH-releasing peptide-2 and GH-releasing hormone in swine

To investigate the effects of high and low somatostatinergic tone on GH-releasing peptide-2 (GHRP-2) and GH-releasing hormone (GHRH)-induced growth hormone (GH) secretion in swine, we examined GHRP-2- and GHRH-induced GH secretion after pretreatment with atropine or pyridostigmine. Pretreatment of swine with atropine (80 µg/kg bodyweight (BW), intravenous (i.v.)) 15 min before i.v. administration of saline, GHRP-2 (30 µg/kg BW), GHRH (1 µg/kg BW) or a combination of GHRP-2 and GHRH, reduced plasma GH area under the curve ( P  < 0.05), completely blocked GH response to GHRH, and attenuated GH response to GHRP-2 and GHRH combined ( P  < 0.05), without affecting GH response to GHRP-2 only. A synergistic effect of GHRP-2 and GHRH was not observed. In contrast, pretreatment of swine with pyridostigmine (100 µg/kg BW, i.v.), under the same pretreatment conditions as above, increased plasma GH concentration ( P  < 0.01), augmented GH response to GHRP-2 ( P  < 0.05), and GHRP-2 and GHRH combined ( P  < 0.05), but did not affect GH response to GHRH. These results suggest that the cholinergic muscarinic agents atropine and pyridostigmine modulate the GH response to GHRP-2 and GHRH, and that GHRP-2 acts antagonistically on the inhibitory effect of somatostatin in swine.     

Response of Biochemical Markers of Bone Metabolism to Exercise Intensity in Thoroughbred Horses

We studied the response of biochemical markers of bone metabolism to exercise intensity in horses. Four horses were walked on a mechanical walker for one week (pre-exercise). Then they performed low-speed exercise on a high-speed treadmill in the first week and medium-speed exercise in the second week and high-speed exercise in the third week of training. We measured two indices of bone resorption, serum hydroxyproline concentration and the urinary deoxypyridinoline/creatinine ratio, and serum osteocalcin (OC) concentration as an index of bone formation. Both indices of bone resorption gradually decreased during the experiment. Serum OC concentration did not change in the first week but was significantly lower in the second and the third weeks compared to in the pre-exercise period and in the first week. These results suggest that the low-speed exercise decreased bone resorption but did not affect bone formation, which possibly results in increasing bone mineral content and strengthening of bones. The high-speed exercise decreased bone formation and bone resorption, i.e., bone turnover was suppressed. The low-speed exercise may be preferable for increasing bone mineral content.