Factors affecting premature chromosome condensation of cumulus cell nuclei injected into rat oocytes

To date, production of cloned rats by somatic cell nuclear transfer (NT) has not yet been successful. Inducing premature chromosome condensation (PCC) of injected cell nuclei in recipient cytoplasm is considered essential for successful mouse cloning by the Honolulu method. In the present study, some factors affecting PCC of rat cumulus cell nuclei injected into rat oocytes were examined. Wistar female rats (young: 4 to 5-week-old, mature: > or =10-week-old) were superovulated by injections of eCG and hCG, and oocytes recovered 14 or 17 h after hCG injection were received with cumulus cell nuclei using piezo-driven micromanipulator. When the oocytes were recovered 14 h post-hCG injection from young rats and the nuclear injection into oocytes was completed within 45 min, PCC was observed in 44-49% of NT oocytes. In the case of oocytes from mature rats, PCC occurred in 11-19% of the NT oocytes. Oocytes recovered 17 h post-hCG injection did not support PCC of the injected nuclei (0-7%) regardless of the donor age. Treatment of oocytes with a neutral cysteine protease inhibitor, N-acetylleucylleucylnorleucinal, slightly increased the incidence of PCC (48 vs 37%). Comparison of rat strains for oocyte donors indicated that proportions of NT oocytes undergoing PCC in Wistar and LEW oocytes (41-46%) were higher than those in Donryu and F344 oocytes (17-25%). Thus, ability of rat oocytes to promote PCC of the injected nuclei is dependent on the characteristics of oocytes, such as age or strain of donor rats, and timing of oocyte recovery.     

Effects of supplemental β‐carotene on colostral immunoglobulin and plasma β‐carotene and immunoglobulin in Japanese Black cows

Data from 26 Japanese Black cows were collected to clarify the effects of supplemental β‐carotene on colostral immunoglobulin (Ig) and plasma β‐carotene and Ig in the cows. Cows were assigned to control or β‐carotene groups from 21 days before the expected calving date to 60 days after parturition. Supplemental β‐carotene was provided at 500 mg/day in the β‐carotene group. Supplemental β‐carotene drastically increased plasma β‐carotene concentrations in the cows from parturition to 60 days after parturition, and plasma β‐carotene concentrations in the control and β‐carotene groups at parturition were 202 and 452 μg/dl, respectively. Supplemental β‐carotene had no effects on plasma IgG₁, IgA or IgM concentrations at parturition. Supplemental β‐carotene increased colostral IgG₁ concentrations in the cows, but colostral β‐carotene, IgA and IgM concentrations were not affected by supplemental β‐carotene. These results indicate that supplemental β‐carotene is effective to enhance colostral IgG₁ concentrations and plasma β‐carotene concentrations in Japanese Black cows.     

Availability of black soldier fly meal as an alternative protein source to fish meal in red sea bream (Pagrus major,Temminck & Schlegel) fingerling diets

Black soldier fly meal (BM) is an outstanding candidate as a fish meal (FM) substitute because it contains relatively high protein and essential amino acids. In this study, we replaced FM in red sea bream diets (Pagrus major) with BM to investigate its effect on growth and feed utilization. Six isonitrogenous and isolipidic experimental diets were prepared by substituting 0%, 20%, 40%, 60%, 80% and 100% FM protein with BM (Control, BM20, BM40, BM60, BM80 and BM100, respectively). After the 8-week feeding trial, final body weight, weight gain, specific growth rate and feed efficiency decreased linearly with increasing dietary BM level (p < 0.05). Apparent digestibility of protein and fat and serum total cholesterol concentration decreased with increasing dietary BM levels (p < 0.05) Final body weight, WG, SGR and FE against dietary BM level had breaking points of 76.2%, 41.7%, 76.5% and 60.0%, respectively, in segmental regression analysis. In conclusion, the results suggest that BM can replace a maximum of 41.7% of FM in the diet of red sea bream without compromising growth performance or feed efficiency for 56 days.     

Groundwater Deterioration Caused by Induced Recharge: Field Survey and Verification of the Deterioration Mechanism by Stochastic Numerical Simulation

Our field survey showed that thequality of shallow groundwater around the KatsuraRiver in the Kyoto Basin was strongly affected by theinfiltration of river water. Furthermore, that thedeterioration of the groundwater in the southern areato the west of the Katsura River may be related to theincrease in groundwater extraction. To clarify themechanism of groundwater deterioration, we havedeveloped a stochastic method to simulate groundwaterflow. The results showed that there was a largereduction in the groundwater level where groundwaterextraction was intense and recharge flowed from theKatsura River to the high extraction areas in thesouthern region. Another simulation showed that if thegroundwater extraction was 10% of the present removalrate, there would be little recharge from the KatsuraRiver into the groundwater and the quality of thegroundwater would be improved. Thus, we conclude thatthe cause of groundwater deterioration is probably dueto the induced recharge of deteriorated river waterfrom the Katsura River.     

Isoflavone-free soy protein prepared by column chromatography reduces plasma cholesterol in rats

To know whether isoflavones are responsible for the hypocholesterolemic effect of soy protein, the effect on plasma cholesterol of isoflavone-free soy protein prepared by column chromatography was examined in rats. Five-week-old male Sprague-Dawley rats were fed cholesterol-enriched AIN-93G diets containing either 20% casein (CAS), 20% soy protein isolate (SPI), 20% isoflavone-free SPI (IF-SPI), 19.7% IF-SPI + 0.3% isoflavone-rich fraction (isoflavone concentrate, IC), or 20% CAS + 0.3% IC for 2 weeks. Plasma total cholesterol concentrations of rats fed SPI and IF-SPI were comparable and were significantly lower than that of rats fed CAS. The addition of IC to the CAS and IF-SPI did not influence plasma cholesterol level. Fecal steroid excretion of the three SPI groups was higher than that of the two CAS groups, whereas the addition of IC showed no effect. Thus, a significant fraction of the cholesterol-lowering effect of SPI in rats can be attributed to the protein content, but the isoflavones and other minor constituents may also play a role.     

The mode of operation of a regulating reservoir for effective use of river flow

In the absence of a reservoir on the downstream side, a river flow that increases temporarily by rainfall cannot be stored and discharged apart from requirements in a command area. This unused water (excess water) occurs to some extent even in a drought year and making effective use of this supply is considered as one of the countermeasures for water shortage. Setting up a regulating reservoir on the upstream side of head works is regarded as one of the methods for making effective use of excess water. The inflow to the river and water delivery loss occurred from rainfall are stored in the regulating reservoir and supplied to the command area on the downstream side as agricultural water. An operational method for the regulating reservoir to effectively utilize the excess water was examined for a canal system by simulation. In an investigated area, there was excess water even in drought years, and making use of it through the regulating reservoir whose capacity is smaller compared to that of a dam can become one of the countermeasures for the drought period. The simulation in this study could estimate quantitatively the effect of the regulating reservoir to make the excess water effective.     

Magnitude of mitral valve closure plays a pivotal role in enhancing the forward blood flow during cardiac massage in dogs with ventricular fibrillation

Motion of mitral valve during cardiac massage was examined using beagle dogs with ventricular fibrillation (n=4). Active compression-decompression cardiac massage (ACD-CM) exhibited greater peak aortic pressure than standard cardiac massage (S-CM), reverse of which was true for peak pulmonary capillary wedge pressure in each animal. Accordingly, peak aortic pressure was greater than peak pulmonary capillary wedge pressure with ACD-CM, whereas its reverse was true with S-CM. Transesophageal echocardiography revealed that mitral valve was incompletely closed with S-CM with showing regurgitation. The valve was more effectively closed during ACD-CM. These results indicate that effective closure of mitral valve during cardiac massage may increase forward blood flow, supporting “cardiac pump theory” rather than “thoracic pump theory” as a principle in dogs.