Prolonged exposure to hyaluronidase decreases the fertilization and development rates of fresh and cryopreserved mouse oocytes

Hyaluronidase is generally used to remove cumulus cells from mouse oocytes before oocyte cryopreservation, intracytoplasmic sperm injection or DNA injection. In general, use of cumulus-free mouse oocytes decreases in vitro fertilizing ability compared with cumulus-surrounded oocytes. The effect of hyaluronidase exposure on the quality of mouse oocytes is not fully understood. Here, we investigated the effect of hyaluronidase exposure time on the fertilization rate of fresh and vitrified mouse oocytes and their subsequent developmental ability in vitro. We found that the fertilization rate decreased with hyaluronidase treatments. This reduction in the fertilization rate following treatment with hyaluronidase was fully reversed by removal of the zona pellucida. In addition, oocytes treated with hyaluronidase for 5 min or longer had a reduced capacity to develop to the morula and blastocyst stage. The survival, fertilization, and developmental rates of vitrified-warmed oocytes were also reduced by longer exposure to hyaluronidase. In conclusion, these results suggest that prolonged exposure to hyaluronidase decreases the quality of mouse oocytes and shorter hyaluronidase treatment times may help achieve a stable and high fertilization rate in fresh and cryopreserved oocytes.     

Temperature dependence of the dynamic viscoelasticity of bases of Japanese cypress branches and the trunk close to the branches saturated with water

With the aim of obtaining findings on the dynamic properties of branches and their bases, as well as their support mechanisms, the present study examined the temperature dependence of the dynamic viscoelasticity of Japanese cypress samples saturated with water to clarify the responses in different regions, and identified factors influencing the characteristics. In the bases of the branches: E′ sharply decreased at approximately room temperature and significantly decreased at around 20 and 60 °C; a peak and shoulder peak of E″ or tan δ were noted at around 20 °C, and there was another peak of tan δ at around 60–80 °C; and mechanical relaxation was noted at around 20 °C and 60–80 °C. On the other hand, in some regions, including the trunks, branches, and their bases, mechanical relaxation was only noted on the high-temperature side. However, boiling treatment with about 12 % weight loss inhibited mechanical relaxation, and there were decreases in E′, E″, and tan δ at approximately room temperature. The bases of the branches of Japanese cypress are considered to develop its elasticity and viscosity to tolerate external stress by accumulating an extract, which enhances the strength of lignin.     

Varietal difference in vitamin C content in the fruit of kiwifruit and other actinidia species

Vitamin C content in the fruit of various cultivars of kiwifruit and other Actinidia species was estimated by determination of L-ascorbic acid and L-dehydroascorbic acid using ion-pair reversed-phase high-performance liquid chromatography. Fruit of A. deliciosa cv. Hayward, the most common commercially available cultivar, contained 65.5 mg/100 g fresh weight (FW) vitamin C. Vitamin C content in A. deliciosa fruit varied from 29 mg/100 g FW to 80 mg/100 g FW. In most cultivars of A. chinensis, vitamin C content in fruit was higher than that of Hayward. In particular, vitamin C content in cv. Sanuki Gold fruit reached more than 3-fold that of Hayward on a weight for weight basis. In A. argutafruit, there was wide variation in vitamin C content, with concentrations ranging from 37 to 185 mg/100 g FW. In cv. Gassan, Issai, and Mitsuko, vitamin C content of the fruit was much higher than that of Hayward. In A. arguta fruit, the ratio of L-ascorbic acid to total ascorbic acid tended to be higher than that of other species.     

Effects of wood kraft pulp feeding on feed digestibility and rumen fermentation of Japanese Black steer in the middle fattening stage

We investigated the effects of wood kraft pulp (KP) feeding on feed digestibility and rumen fermentation of Japanese Black (JB) steer in the middle fattening stage. The feeding experiment was carried out a replicated 3 × 3 Latin square design using six JB steers (16.2 ± 0.8 months of age). Steers were fed rice straw and three concentrated feeds: commercial formula feed (control), formula feed containing 7.5% KP on a dry matter (DM) basis (7.5% KP), and formula feed containing 15% KP on a DM basis (15% KP). Feed digestibility and rumen fermentation parameters were recorded for 3 days during the measurement period, after an adaptation phase of 11 days. There was no significant difference in DM intake per day among diets. A slightly higher digestibility of neutral detergent fiber was observed in steers fed 15% KP, compared with those fed other diets (P = 0.059). The butyric acid ratio in rumen fluid was higher in steers fed KP diets than in steers fed control (P = 0.083). The average ruminal pH was unaffected by KP diets. This study suggests that replacing 15% of concentrated feed with KP has little effect on feed digestibility and rumen fermentation in JB fattening steer.     

Ability of alphas-Casein to suppress the heat aggregation of ovotransferrin

The effects of alphas-casein on heat aggregation of ovotransferrin (OT) were studied by heating at 80 degrees C for 20 min in 10 mM phosphate buffer, pH 7.0. The heat interactions between alphas-casein and OT were followed by turbidity development and polyacrylamide gel electrophoresis. We found that alphas-casein can effectively suppress the heat-induced aggregation of heat-labile OT. The suppressive ability of alphas-casein was reduced by the presence of NaCl on heating. Dephosphorylated alphas-casein had less ability to suppress the aggregation of OT than native alphas-casein. Our results indicate that alphas-casein interacts with the heat-denatured OT through its exposed hydrophobic surface and phosphoserine residue. Such interactions seem to be important in helping to suppress the aggregation of heated OT. The suppressive effects of alphas-casein on heat aggregation of OT would be partially ascribed to the formation of transparent gel from egg white by the addition of alphas-casein.     

Atomic force microscopy studies on heat-induced gelation of curdlan

Heat-induced gelation of a cold-water insoluble polysaccharide, Curdlan, was investigated using atomic force microscopy (AFM). Curdlan dissolved into NaOH aqueous solutions exhibited a spectral transition around 0.2 mol/L NaOH, which is an indicative of conformational transitions from a single helix at a lower alkali concentration to a disordered chain at a higher concentration. Nevertheless, AFM images of Curdlan solubilized in 0.01 mol/L NaOH revealed the presence of heterogeneous supramolecular assemblies of Curdlan: the majority of the molecules were in the form of microfibrils, the lengths of which were on the order of micrometers and the cross-sectional heights of which were approximately 2-3 nm, whereas single molecular chains, partially dissociated from these microfibrils, were also observed. Heating such a sol resulted in the formation of densely cross-linked microgel networks. Heat-induced gelation of Curdlan appears to be initiated by partial dissociation of single chains from supramolecular microfibrils and followed by cross-linking of microfibrils via hydrophobic interactions among these partially dissociated chains.     

Protective effects of dietary chamomile tea on diabetic complications

Matricaria chamomilla L., known as "chamomile", has been used as an herbal tea or supplementary food all over the world. We investigated the effects of chamomile hot water extract and its major components on the prevention of hyperglycemia and the protection or improvement of diabetic complications in diabetes mellitus. Hot water extract, esculetin (3) and quercetin (7) have been found to show moderate inhibition of sucrase with IC50 values of 0.9 mg/mL and 72 and 71 microM, respectively. In a sucrose-loading test, the administration of esculetin (50 mg/kg body weight) fully suppressed hyperglycemia after 15 and 30 min, but the extract (500 mg/kg body weight) and quercetin (50 mg/kg body weight) were less effective. On the other hand, a long-term feed test (21 days) using a streptozotocin-induced rat diabetes model revealed that the same doses of extract and quercetin showed significant suppression of blood glucose levels. It was also found that these samples increased the liver glycogen levels. Moreover, chamomile extract showed potent inhibition against aldose reductase (ALR2), with an IC50 value of 16.9 microg/mL, and its components, umbelliferone (1), esculetin (3), luteolin (6), and quercetin (7), could significantly inhibit the accumulation of sorbitol in human erythrocytes. These results clearly suggested that daily consumption of chamomile tea with meals could contribute to the prevention of the progress of hyperglycemia and diabetic complications.     

Mortality processes of hatchery‐reared Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) larvae in relation to their piscivory

In mass culture of Pacific bluefin tuna Thunnus orientalis, yolk‐sac larvae of other species are fed as a major prey item to tuna larvae from 7 to 8 mm in total length. Marked growth variations in tuna larvae are frequently observed after feeding of yolk‐sac larvae, and this variation in the growth of tuna larvae is subsequently a factor leading to the prevalence of cannibalistic attacks. To elucidate details of the mortality process of hatchery‐reared tuna larvae after the initiation of yolk‐sac larvae feeding, we compared the nutritional and growth histories of the surviving (live) tuna larvae to those of the dead fish, found dead on the bottom of the tank, as direct evidence of their mortality processes. Cause of mortality of tuna larvae 3 and 5 days after the initiation of feeding of yolk‐sac larvae was assessed from nitrogen stable isotope and otolith microstructure analyses. Stable isotope analysis revealed that the live fish rapidly utilized prey fish larvae, but the dead fish had depended more on rotifers relative to the live fish 3 and 5 days after the initiation of feeding of yolk‐sac larvae. The growth histories based on otolith increments were compared between the live and dead tuna larvae and indicated that the live fish showed significantly faster growth histories than dead fish. Our results suggest that fast‐growing larvae at the onset of piscivory could survive in the mass culture tank of Pacific bluefin tuna and were characterized by growth‐selective mortality.     

Phenotypic and genotypic characterization of Actinobacillus suis sensu stricto isolated from a dairy calf

The species of the genus Actinobacillus have so far been associated with specific animal hosts, and A. suis sensu stricto, an opportunistic pathogen of swine, is rarely isolated from ruminants. We describe here the isolation of A. suis sensu stricto from a newborn calf that died on a dairy farm in Japan. Identification of the isolate was performed by phenotypic and genotypic characterization, with the latter consisting of nucleotide sequence analyses of the 16S rRNA gene plus three housekeeping genes, rpoB, infB and recN.