Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Hyperglycaemia in transition dairy cows: Effects of lactational stage and conjugated linoleic acid supplementation on glucose metabolism and turnover

Supplementing conjugated linoleic acid ( CLA ) is supposed to spare glucose due to the milk fat‐depressing effect of the trans ‐10, cis ‐12 CLA isomer, and allows repartitioning nutrients despite an energy deficiency in early lactation. However, there is still a lack of knowledge in terms of the dynamic pattern of the glucose turnover in transition dairy cows. We hypothesized that dairy cows supplemented with CLA have an altered rate of glucose turnover and insulin sensitivity during early lactation. We conducted three consecutive hyperglycaemic clamps (HGC ) in weeks ?2, +2 and +4 relative to parturition in Holstein cows supplemented daily either with 70 g of lipid‐encapsulated CLA (6.8 g trans ‐10, cis ‐12 and 6.6 g of the cis ‐9, trans ‐11 CLA isomer; CLA ; n  = 11) or with 56 g of control fat ( CON ; n  = 11). From week ?3 up to week +4 relative to parturition, milk yield and dry matter intake (DMI ) were recorded daily, while body weight (BW ) and milk composition were obtained once weekly. Blood samples were taken once weekly and every 30 min during the HGC . Plasma was analysed for concentrations of glucose, fatty acids (FFA ), beta‐hydroxybutyrate (BHB ), insulin, triglycerides and cholesterol. The CLA supplementation did not affect performance and metabolic parameters except for BHB and cholesterol. Furthermore, insulin concentrations and insulin sensitivity were affected by treatment. During the HGC in early lactation, insulin response was lower and decrease in FFA and BHB greater compared with the HGC in week ?2 although glucose target concentration achieved during the steady‐state period was similar for all three HGC . Our findings in terms of insulin and cholesterol suggest that body reserves are preserved through CLA feeding without restraining animal's performance. Furthermore, CLA effects on cholesterol and triglyceride concentrations indicated beneficial effects on hepatic lipid export contributing to an improved efficiency of prevailing metabolites in circulation.     

Patterns of SSR variation in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seeds under ex situ genebank storage and accelerated ageing

Maintaining seed viability and germplasm integrity is a challenging task in conservation of plant genetic resources, as seeds under storage will lose viability and genetic changes will occur. Attempt was made to analyze the patterns of genetic changes in wheat germplasm under ex situ genebank storage and accelerated ageing treatments. A set of 16 naturally aged wheat accessions under ex situ genebank storage since 1994 were sampled. Four recently regenerated wheat accessions were selected, four random seed samples were chosen from each accession, and three of them were exposed to three different accelerated ageing treatments. These 32 seed samples in two germplasm sets displayed a range of germination rates from 4 to 98 %. Thirty-seven microsatellite markers representing 21 wheat chromosomes were applied to screen 12 seeds of each sample and 449 SSR alleles were scored. Large SSR variation was found in each germplasm set. There was 73.1 % of the total SSR variation present among the naturally aged samples and 78.2 % present among the accelerated ageing samples. Several analyses for genetic association consistently revealed no clear genetic separations among samples of high or low germination rates in both germplasm sets. Samples under different accelerated ageing treatments did not show much genetic differentiations from the original sample of each accession. Mantel tests revealed non-significant associations between SSR variability and sample germination rates for both germplasm sets. These findings are useful for understanding seed deterioration under different ageing conditions and suggest that genome-wide SSR variability may not provide sensitive markers for the monitoring of wheat seed viability.     

不同质地土壤玉米秸秆还田配施腐熟剂效应的研究

为研究鉴选不同质地土壤秸秆还田适宜的秸秆腐熟剂,于2017—2018年在西辽河平原灌区中壤土和砂壤土秸秆还田配施人元腐熟剂和中农绿康腐熟剂,以秸秆还田不施腐熟剂为对照,共设置6个处理,测定玉米秸秆腐解率、玉米产量、玉米根系特性和土壤化学性状,研究不同秸秆腐熟剂对玉米产量及土壤特性的影响。结果表明:不同质地土壤2种腐熟剂秸秆腐解率间无显著差异;中壤土2年各处理玉米产量及根系特性、土壤化学性状均表现为人元腐熟剂中农绿康腐熟剂秸秆还田不施腐熟剂,且中农绿康腐熟剂和人元腐熟剂显著高于秸秆还田不施腐熟剂,而2种腐熟剂处理之间无显著差异;砂壤土2年各处理玉米产量及根系特性、土壤化学性状均表现为中农绿康腐熟剂人元腐熟剂秸秆还田不施腐熟剂,且中农绿康腐熟剂显著高于人元腐熟剂和秸秆还田不施腐熟剂。2年不同质地土壤中配施腐熟剂在根系特性、土壤化学性状方面表现为砂壤土配施中农绿康腐熟剂效果优于人元腐熟剂,而中壤土配施2种腐熟剂效果无显著性差异。     

芦荟对肉仔鸡生长性能和免疫功能的影响

将360只1日龄雄性AA肉仔鸡随机分成6个组,每组设6个重复,每个重复10只.Ⅰ组为空白对照组,饲喂基础日粮,Ⅱ-Ⅵ组分别在基础日粮中添加600 mg.kg-1芦荟粉、600 mg.kg-1芦荟水提取物、600 mg.kg-1芦荟醇提取物、600mg.kg-1芦荟水提取物+600 mg.kg-1芦荟醇提取物和150 mg.kg-1金霉素.结果表明:Ⅲ组3、6周鸡的体增重均显著高于Ⅰ组(P<0.05);Ⅲ-Ⅳ组的料肉比与Ⅰ组比较均极显著降低(P<0.01),Ⅲ组与Ⅵ组比较显著降低(P<0.05);Ⅱ-Ⅴ组3周鸡的脾脏指数与Ⅰ组相比,差异均显著提高(P<0.05);Ⅲ组3周鸡的胸腺指数显著高于Ⅰ组(P<0.05);Ⅱ-Ⅴ组3周鸡的自然杀伤细胞的杀伤活性均极显著高于Ⅰ组(P<0.01),Ⅱ-Ⅴ组6周鸡均极显著高于Ⅰ、Ⅵ组(P<0.01).可见芦荟粉、水提取物、醇提取物和提取物混合组均能提高肉仔鸡的生产性能和免疫功能,其中水提取物组的效果最佳.