Growth of bovine oocyte-granulosa cell complexes cultured individually in microdrops of various sizes

In mammalian embryo culture, the embryo:medium volume ratio can substantially affect embryo developmental performance. In the present study, we tested the possibility of improving the growth of bovine oocytes by reducing the medium volume, from a typical volume used in mouse follicle culture to a minimum possible level. A total of 282 complexes, each containing a growing oocyte 87-100 mum in diameter, were individually placed in microdrops of 2, 5, 10 or 20 microl and cultured for 13 days in a modified TCM-199 supplemented with 4% polyvinylpyrrolidone (molecular weight: 360 kDa). Oocyte diameter was measured every other day to trace the growth of each oocyte. Half the medium was replaced every other day or every day, and comparison revealed that daily replacement was more favorable for culture of these microdrops. The highest survival rate, 95%, occurred in the 20-microl microdrops, where most oocytes continued to grow throughout the culture period. In comparison, in the 5- and 10-microl microdrops, more oocytes died, and growth slowed towards the end of culture. In the 2-microl microdrops, which had the highest death rate, growth virtually ceased after 9 days. The surviving oocytes were usually accompanied by a characteristic dome-like structure of the granulosa cell mass, except in the 2-microl microdrops. In conclusion, the 20-microl microdrops allowed oocyte growth at an acceptable level, and any further reduction of the volume only had a negative impact on oocytes.     

Evaluation of the temperature and relative humidity preferences of the western dry-wood termite <Emphasis Type="Italic">Incisitermes minor</Emphasis> (Hagen) using acoustic emission (AE) monitoring

Twenty-four different combinations of six temperatures (15°, 20°, 25°, 30°, 35°, and 40°C) and four relative humidity (RH) (60%, 70%, 80%, and 90%) conditions were used for pseudergates of the western dry-wood termite Incisitermes minor (Hagen). The feeding activities of the termites were monitored by the detection of generated acoustic emission (AE) events from feeder wood blocks in a test chamber. Temperature and RH showed independent and interactive significant effects on the feeding activity of I. minor. The optimal temperature and RH conditions for the feeding activities were 35°C and 70%, respectively, and the optimal combinations were 35°C-70% and 35°C-80% with an exceptionally higher feeding activity at the combination condition of 30°C-70%.     

农村电网优化电压制式及经济供电半径的研究

随着中国农村电气化的发展,农网的用电负荷和用电量与日俱增,现行的电压制式已不能满足需求,如何选取更好的电压等级制式方案成为迫切需要解决的问题。该文在对电网结构模型进行合理设计的基础上,通过对电网经济参数、费用的计算,建立以单位供电面积年费用最小为目标函数,以电网各电压等级供电半径为优化变量的有约束非线性规划模型。应用该模型优化所得的目标值对不同电压等级制式方案进行经济比较,研究结果表明未来中国农网电压制式应简化电压等级,中压配电电压应发展到20 kV及35 kV,农网供电半径应根据经济供电半径的参考取值进     

养殖密度对圆形循环水养殖池自清洗能力的影响

循环水养殖具有养殖密度大、环境污染低、经济效益高的优点，是重要的水产养殖模式。然而，如何快速高效地排出养殖池内的残饵粪便等污物，降低其对水质的影响是循环水养殖模式中面临的首要问题。该研究采用物理试验研究鱼类养殖密度对圆形循环水养殖池的水动力特性及污物运动汇集的影响，揭示不同流量驱动下养殖密度与养殖池自清洗能力的响应关系。结果表明：提高鱼类养殖密度会降低养殖池内整体流场的平均流速v_avg，衰减幅度在0.05 m/s（25％）以内，并提高水中阻力系数C_t；鱼类游动引起的湍流能够导致池内污物再悬浮，有助于污物排出；集污时间同时受到养殖密度和流量的影响，9.8 L/min进水流量下集污时间都在5 min以内；进水流量为6.54 L/min时，养殖密度从0提高到6.2 kg/m³，湍流强度提高2.4倍，集污时间减少了40 min以上。因此，设计循环水养殖系统时需要综合考虑进水流量和预期养殖密度对养殖池自清洗性能的综合影响。研究结果可为圆形循环水养殖池的设计和日常管理提供参考。     

Interactions between the oocyte and surrounding somatic cells in follicular development: lessons from in vitro culture

Mammalian oogenesis occurs concomitantly with folliculogenesis in a coordinated manner in the ovaries. In vitro growth (IVG) culture systems of the oocytes have been developed as a new technology for utilizing incompetent oocytes in the ovary as a source of mature oocytes as well as for studying oogenesis, folliculogenesis, and oocyte-somatic cell interactions. The results of IVG experiments have suggested that direct association of oocytes and surrounding granulosa cells supports oocyte viability and growth through the gap junctions, which are efficient conduits for low molecular weight substances. It has been revealed that granulosa cells metabolize some molecules which are in turn transported into the oocytes. IVG systems have also provided evidence that FSH promotes the development of follicles at secondary or later stages by its stimulation of proliferation and differentiation of granulosa cells, and perhaps by its anti-apoptotic effects. In addition, interactions between granulosa cell-derived KIT ligands and oocyte KIT receptors have been suggested as initiating oocyte growth and follicular development. Furthermore, recent findings suggest there are growth factors derived from oocytes such as GDF-9 and BMP-15. With such factors, oocytes participate in follicular development by regulating the differentiation of surrounding somatic cells. These bidirectional communications between oocytes and somatic cells are important for oocyte growth and follicular development. IVG systems should provide further information regarding oogenesis and folliculogenesis in the ovary.     

Phage conversion of exfoliative toxin A in Staphylococcus aureus isolated from cows with mastitis

An exfoliative toxin produced by Staphylococcus aureus is the causative agent of staphylococcal scalded-skin syndrome (SSSS) in young children. Recently, we reported that only few isolates of S. aureus from bovine mastitis contained the eta gene encoding exfoliative toxin A (ETA) and produced ETA in vitro. In this study, we isolated temperate phages from two ETA-positive bovine isolates of S. aureus by treatment with mitomycin C. Polymerase chain reaction (PCR) assay of the phage genomes suggested that the temperate phages carried the structural gene for ETA. Moreover, the nucleotide sequence analysis of the PCR products revealed that the eta gene was located very close to an amidase gene on the phage genomes. The nucleotide sequence for the amidase gene of the bovine phage (bovine phi ETA) differed at nine positions from that of the amidase gene of phi ETA from a human isolate reported by Yamaguchi et al. [Mol. Microbiol. 38 (2000) 694], suggesting that eta-converting phages are heterogeneous. Bovine phi ETA had a head with a hexagonal outline and a non-contractile and flexible tail. Bovine phi ETA was able to lysogenize ETA-negative bovine isolates of S. aureus, and the lysogenized S. aureus isolates had the ability to produce ETA. These results suggest the possibility of horizontal transmission of the eta gene by temperate bacteriophages among bovine isolates of S. aureus.     

Age related decreases in neural sensitivity to NaCl in SHR-SP

To determine whether neurophysiological taste responses of young and old rats are different, recordings were made from the whole chorda tympani nerve which innervates taste buds on the anterior tongue. SHR-SP (Stroke-Prone Spontaneously Hypertensive Rats) in two age groups were studied. Chemical stimuli included single concentrations of 250 mM NH(4)Cl, 100 mM NaCl, 100 mM KCl, 500 mM sucrose, 20 mM quinine-hydrochloride, 10 mM HCl, 10 mM monosodium glutamate (MSG), 10 mM L- glutamic acid (L-Glu) and an NaCl concentration series. The magnitude of the neural response (response ratio) was calculated by dividing the amplitude of the integrated response by the amplitude of the spontaneous activity that preceded it. Substantial neural responses to all chemicals were obtained at both ages. The responses to KCl, sucrose, quinine-hydrochloride, HCl, monosodium glutamate (MSG) and glutamic acid (Glu) did not change with age, but the response to NaCl did decrease significantly. The profile of the response/concentration function for NaCl differed with age. In particular, the responses to solutions more concentrated than 100 mM NaCl were significantly weaker in aged than in young SHR-SPs. We also observed that recovery from amiloride treatment on the tongue of SHR-SPs was faster in aged rats than in young ones, suggesting that there is some functional difference in the sodium-specific channels on the taste cell. These results suggest that aged SHR-SP may be less able than young SHR-SPs to discriminate among higher concentrations of NaCl solutions.     

Improvement of the gelling properties of meat emulsion gel by the addition of porcine sarcoplasmic proteins

The effects of porcine sarcoplasmic proteins (SP) on the physicochemical properties of meat emulsion gel were examined. Meat emulsion was prepared from water‐washed pork meat (WWM), corn oil and SP. Whole SP (W‐SP) enhanced the breaking stress of the WWM emulsion gel as well as other animal proteins in the presence of 0.2 mol/L NaCl. The breaking stress of the emulsion with W‐SP increased with an increase in corn oil content. Furthermore, this tendency was more noticeable at a lower NaCl concentration (0.15 mol/L) rather than at 0.45 mol/L NaCl. Ammonium sulfate (AS) treatment fractionated W‐SP into three portions (SP‐f1, SP‐f2 and SP‐f3), which were the precipitates at 0–50% and 50–75% AS saturation and the supernatant at 75% AS saturation, respectively. The fractions SP‐f2 and SP‐f3 increased the gel strength more than W‐SP. In particular, the fraction SP‐f3 increased the gel strength approximately 10‐fold compared to the control. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis analysis showed that SP‐f3 had several kinds of proteins and a main protein with a molecular mass of 35 kDa, which corresponded to glyceraldehyde 3‐phosphate dehydrogenase. These results indicate that the influence of SP should not be ignored when processing of low‐salt meat products and the fraction SP‐f3 has a gel‐enhancing factor for myofibrillar proteins.     

Decreases of apolipoprotein B-100 and A-I concentrations and induction of haptoglobin and serum amyloid A in nonfed calves

Reduced feed intake near parturition is suggested to be one of the major causal factors for the development of fatty liver in cows, and nonfeeding has been used as an experimental model for fatty liver. In cows with fatty liver, concentrations of lipoprotein lipids and proteins are decreased. In addition, the acute-phase protein haptoglobin is induced. The purpose of the present study was to examine whether the decrease of lipoprotein concentrations and the induction of acute-phase proteins were similarly reproduced by non-feeding. Holstein female calves (n=5) were nonfed for 3 days and thereafter refed. Serum concentrations of nonesterified fatty acids and beta-hydroxybutyric acid were initially increased by the nonfeeding, and followed by decreases in concentrations of cholesteryl esters, phospholipids, apolipoprotein (apo) B-100 and apoA-I. The apoC-III concentration was not distinctly decreased. Haptoglobin and serum amyloid A were induced during the nonfeeding and refeeding process. Haptoglobin was distributed in different proportions in the high-density lipoprotein, very high-density lipoprotein and the lipoprotein-deficient fractions, whereas almost all serum amyloid A was associated with the high-density lipoprotein fraction. These results suggest that the decreases in lipoprotein concentrations and induction of acute-phase proteins found in cows with fatty liver and those with fatty liver-related diseases such as ketosis are primarily due to the reduced feed intake near parturition.     

Production efficiency and telomere length of the cloned pigs following serial somatic cell nuclear transfer

The aim of the present study was to examine the production efficiency of cloned pigs by serial somatic cell nuclear transfer (SCNT) and to ascertain any changes in the telomere lengths of multiple generations of pigs. Using fetal fibroblasts as the starting nuclear donor cells, porcine salivary gland progenitor cells were collected from the resultant first-generation cloned pigs to successively produce second- and third-generation clones, with no significant differences in production efficiency, which ranged from 1.4% (2/140) to 3.3% (13/391) among the 3 generations. The average telomere lengths (terminal restriction fragment values) for the first, second and third generation clones were 16.3, 18.1 and 20.5 kb, respectively, and were comparable to those in age-matched controls. These findings suggest that third-generation cloned pigs can be produced by serial somatic cell cloning without compromising production efficiency and that the telomere lengths of cloned pigs from the first to third generations are normal.