Vitrification of mouse two-cell and blastocyst stage embryos in simplified closed system using either a hemi-straw or a hollow fiber device

Two-cell stage and blastocyst stage mouse embryos were equilibrated in a medium containing 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 8–15 min. Vitrification was performed in a medium containing 0.5 M sucrose and either 15% EG + 15% DMSO, 17.5% EG + 17.5% DMSO, or 20% EG + 20% DMSO for 30 s. They were then placed either on a hemi-straw (HS) or a hollow fiber vitrification (HFV) device and vitrified by cooled air inside a 0.5-ml straw. In two-cell embryos, a 100% survival rate was obtained from all groups except the 20% HS group (P > .05). All vitrified two-cell groups showed similar rates of blastocyst development to that of fresh control group (P > .05), except 17.5% and 20% HFV groups, which were significantly lower than the other groups (P < .05). In the blastocyst embryos, the HFV groups were divided into two subgroups (non-collapsed; HFV-NC and collapsed; HFV-C blastocyst). Re-expansion rate in 15% HFV-NC, 17.5% HFV-NC, and 15% HFV-C groups was reduced (P < .05), whereas the rest were similar to control. In conclusion, we established a simplified, reliable, and closed system for HFV vitrification applying hemi-straw, which does not require skilled practitioners.     

The effects of density and artificial substrate on intensive shrimp Litopenaeus vannamei nursery production

Recirculating aquaculture systems (RAS) can be installed indoors, allowing year-round production of tropical animals in nearly any climate. A nursery phase is commonly used in Litopenaeus vannamei production since it allows for enhanced biosecurity and better quantification of animals while reducing space requirements. However, it is unclear whether animal density and inclusion of artificial substrate may improve shrimp performance during the nursery phase. In this experiment, we compared shrimp production parameters in two stocking densities with or without the use of an artificial substrate by creating four treatments: low-density LD; 1500 PL/m⁻³, low-density with substrate LDS, high-density HD; 3000 PL/m⁻³), and high-density with substrate (HDS). The LDS and HDS treatments included 0.46-m² of high-density polyethylene 2.5-cm mesh as a substrate, which increased the tank surface area by 21 %. Each treatment was randomly assigned to four 160-l culture tanks, each with a biofilter. The shrimp had an initial weight of 4 mg and were grown for 50 days. The low-density treatments had significantly higher dissolved oxygen (DO) and pH than the high-density treatments (P ≤ 0.001). Specifically, LDS had the highest DO and pH followed by the LD, HD, and HDS treatments, respectively. High-density treatments had significantly higher NO₂-N levels than low-density treatments during week 2 of the experiment when an unusually high concentration of nitrite was observed. FCR was significantly lower in both low-density treatments than in high-density treatments. At harvest, the total biomass (kg m⁻³) was significantly higher in high-density treatments than in low-density treatments (P ≤ 0.001), and the HDS treatment had a significantly greater biomass output than HD. Producers should consider artificial substrate and higher densities during nursery production to maximize shrimp production; however, the effects on water quality should also be taken into account.     

昆明鼠卵母细胞的孤雌发育及对其干扰核移植试验的预防措施

本文着重研究了在昆明鼠细胞核移植过程中，卵母细胞孤雌发育的激活原因、形成过程，并提出了一些防止其干扰细胞核移植的措施。     

Genotype evaluation,muscle histopathology and ultrastructure in a Duchenne muscular dystrophy animal model

AIM: To evaluate the genotype , muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.     

Passive protection of mice with antiserum to neuraminidase fromPasteurella multocida serotype A:3

Antiserum to a partially purified neuraminidase fromPasteurella multocida, type A:3, was adsorbed with protease-digestedP. multocida type 3 lipopolysaccharide (LPS) to remove LPS immunoreactivity. The LPS-adsorbed antineuraminidase caused a 77% reduction in the neuraminidase activity of homologousP. multocida in anin vitro enzyme neutralization test. All 14 mice passively immunized with the adsorbed antineuraminidase were protected against challenge infection with homologousP. multocida in a mouse protection test. Ten out of 14 mice in one group that received antisera containing antibodies to both neuraminidase and LPS were protected. In contrast, only 1 out of 14 mice that were immunized with pre-immune serum survived the challenge. These results suggest that antiserum toP. multocida neuraminidase was, at least partly, responsible for the protection observed in this study. Neuraminidase may be one of the immunogenic protective proteins present in aqueous extracts ofPasteurella multocida.     

切割取样对胚胎发育的影响

体视显微镜下,采用徒手持金属刀片和自制的玻璃切割针分别对小鼠和牛胚胎进行切割取样,并对取样后的胚胎进行体外培养48h,其发育率分别为76.7%(46/60)和80%(16/20),两者差异不显著(P>0.05);奶牛新鲜胚胎切割取样后,胚胎移植妊娠率47.1%(8/17),比对照组妊娠率59.0%(23/39)差异显著(P<0.05)。冷冻-解冻胚胎切割取样后移植妊娠率42.9%(6/17)虽然也比对照组50%低,但是差异不显著(P>0.05)。     

提高中锰白口铸铁韧性的研究

为改善中锰白口铸铁的韧性以扩大其应用范围,本文探讨了C, Si, Mn等合金元素、变质处理和热处理工艺对中Mn白口铸铁组织和性能的影响.     

黑龙江省西部地区重瓣白玫瑰引种栽培及繁育技术

经在黑龙江省齐齐哈尔地区对重瓣白玫瑰引种栽培及繁育技术研究表明：该树种　１９９７～２０００年的平均抗寒指数为　２．７，冻害小于Ⅲ级，枝条冻伤长度小于　５０％，能正常开花，可以在此安家落户。同时对其形态特征、主要物候期、繁育技术和抚育管理作了介绍。     

农户防鼠密闭储粮袋

筛选出了防鼠效果良好的微胶囊化的放线菌酮及其最佳使用浓度，并用其制出了用维纶聚氯乙烯双面涂塑为材料的具有良好防鼠性能、防潮、密封性好，便于防治害虫、重量轻、容量大、成本低的农户储粮袋。