设置创新创业教育专业——破解创新创业教育改革困境的范式转换

培养创新人才是不同发展阶段、不同形态社会对教育的基本诉求。通过对我国部属农林高校创新创业教育改革实施方案文本分析可知,在开展创新创业教育改革发展目标方面高度认同,在组织机构设置方面高度重视,在选择举措方面高度完备。同时存在目标定位无特色又高度雷同,相关部门间协同育人机制缺失以及对发力点缺乏明晰的判断等问题。推进范式转换,设置创新创业教育专业、建立创新创业教育监督执行机构成为当下破解创新创业教育改革困境的重要选择。     

不同单酶制剂混合检测对比研究

试验以木聚糖酶、纤维素酶、淀粉酶和酸性蛋白酶为对象,首先研究了酸性蛋白酶分别和其他3种单酶进行混合后酶活测定值变化情况,结果表明,酸性蛋白酶对其他3种单酶的检测结果没有影响;试验第二部分研究了4种单酶整体混合在一起后各单酶测定值的变化情况,结果表明,木聚糖酶测定值出现很大偏高,其他3种单酶测定结果未出现显著偏差;进行进一步论证试验表明,淀粉酶可能是使木聚糖酶测定结果偏高的原因。     

光照强度对曙箣竹生长及观赏形态影响

以曙箣竹为研究材料,通过不同光照条件下盆栽曙箣竹的生长状况及生理指标分析,探讨光照对曙箣竹生长及观赏形态的影响。研究结果表明:(1)在温室全光照下曙箣竹新增叶量达到最大值,为11片,25%遮光下增量最少仅有3片;在50%光照条件下曙箣竹的平均主秆增长量较大为4.48 cm,25%遮光下增长量最低为1.63 cm;(2)叶片叶绿素总量、叶绿素a、叶绿素b质量分数均随遮光强度的增加而增长,以25%光照叶绿素总量、叶绿素a、b为最高,分别为全光照处理的106.3%、101.9%、110%;叶绿素a/b比值下降,以25%遮光降幅最大,与对照差异显著;(3)随着时间的延长,50%光照下SOD活性呈先增大后减小的现象,25%光照和全光照的SOD活性均呈逐渐增大;50%光照和全光照的POD活性变化呈现相反的现象,前者逐渐减小,后者缓慢增加,25%光照下则是先增大后减小。综合测定指标与观察结果显示:当光照强度为50%不仅促进曙箣竹的生长还有利于观赏性的表达。     

Evidence for functional G-coupled protein receptors 43 and 120 in subcutaneous and intramuscular adipose tissue of Angus crossbred steers

We conducted 3 independent experiments to demonstrate functional G-coupled protein receptor 43 (GPR43) and GPR120 in bovine intramuscular (i.m.) and subcutaneous (s.c.) adipose tissues. We hypothesized that media volatile fatty acids and long-chain fatty acids would affect cAMP-activated protein kinase-alpha (AMPKα) protein expression and cAMP concentrations differently in i.m. and s.c. adipose tissue. Experiment 1: oleic acid (18:1n-9) decreased phosphorylated AMPKα protein (p-AMPKα) and the p-AMPKα/AMPKα protein ratio in i.m. preadipocytes, increased the p-AMPKα/AMPKα protein ratio in bovine satellite cells, and had no effect in s.c. preadipocytes. Experment 2: ex vivo explants from the 5th to 8th longissimus thoracic rib muscle section of Angus crossbred steers were cultured 48 hr in media containing 0.25 µM ciglitizone, 5 mM glucose, and 5 mM acetate, in the absence or the presence of 100 µM oleic acid. Oleic acid increased acetate incorporation into fatty acids and GPR43 gene expression in i.m. adipose tissue (P < 0.05), but oleic acid had no effect on fatty acid synthesis or GPR43 expression in s.c. adipose tissue. Experiment 3: fresh s.c. and i.m. adipose tissue from the 5th to 8th longissimus thoracic rib muscle section of Angus crossbred steers was transferred immediately to 6-well culture plates containing 3 mL of KHB/Hepes/5 mM glucose. Samples were preincubated with 0.5 mM theophylline plus 10 μM forskolin for 30 min, after which increasing concentrations of acetate or propionate (0, 10⁻³, 10^−2.3, and 10⁻³ M) in the absence or the presence of 100 μM oleic acid or 100 µM palmitic acid (16:0) were added to the incubation media. Acetate had no effect on forskolin-stimulated cAMP production in s.c. adipose tissue but decreased cAMP in i.m. adipose tissue (P < 0.05); this indicates a functional GPR43 receptor in i.m. adipose tissue. The combination of 10⁻² M acetate and oleic acid decrease cAMP production in s.c. adipose tissue, consistent with GPR120 receptor activity, but oleic acid and palmitic acid attenuated the depression of cAMP production caused by acetate in i.m. adipose tissue. Palmitic acid depressed cAMP production in s.c. adipose tissue, and increased cAMP production in i.m. adipose tissue (P < 0.05). Propionate had no effect on cAMP production in s.c. or i.m. adipose tissue. These results provide evidence for functional GPR43 receptors in i.m. adipose tissue and GPR120 receptors in s.c. adipose tissue, both of which would suppress lipolysis.     

Effects of dietary Bacillus coagulans and yeast hydrolysate supplementation on growth performance,immune response and intestinal barrier function in weaned piglets

The present study investigated the effects of Bacillus coagulans and yeast hydrolysate supplementation on growth performance, immune response and intestinal barrier function of weaned piglets. Twenty-four weaned piglets with an average body weight (BW) of 6.89 ± 0.15 kg were divided into four diets for 28 days. The treatments were basal diet (control), basal diet supplemented with antibiotic (20 mg/kg colistin sulphate and 40 mg/kg bacitracin zinc, AT), probiotics (400 mg/kg Bacillus coagulans ≥5 × 109 CFU/g, BC) or yeast hydrolysate (5000 mg/kg yeast hydrolysate, YH). Average daily gain (ADG) and average daily feed intake (ADFI) were improved by AT and YH diets (p < 0.05), while BC diet only increased ADG (p < 0.05). The complement 3 (C3), lysozyme (LZM) and tumour necrosis factor-α (TNF-α) concentrations in serum were increased in BC diet (p < 0.05). Feeding AT and YH caused the increase of jejunal villus height (p < 0.05), and a higher ratio of villus height/crypt depth was observed in AT, BC and YH groups (p < 0.05). The mRNA expression of zonula occludens-1 (ZO-1) in jejunal mucosa was up-regulated by AT, BC and YH diets (p < 0.05). Dietary AT, BC or YH inclusion decreased the interleukin-1β (IL-1β) concentration and TNF-α mRNA expression (p < 0.05), and YH supplementation even down-regulated toll-like receptor 4 (TLR4) and CD14 expressions (p < 0.05). In summary, the dietary administration of BC or YH both improves growth performance through promoting the intestinal barrier function, indicating both of them can serve as potential alternatives to antibiotics growth promoters for the piglet production.     

口蹄疫病毒A型竞争ELISA抗体检测试剂盒在流行病学调查中的应用

为评价口蹄疫病毒A型竞争ELISA(cELISA)抗体检测试剂盒在流行病学调查中的应用前景,对2017年从福建省三明市采集的336份黄牛、奶牛、羊和猪血清样品,用A型cELISA抗体检测试剂盒进行抗体检测。结果显示,92份黄牛血清、92份羊血清、92份猪血清、60份奶牛血清的A型抗体阳性率分别为13.04%、11.96%、20.65%、86.67%。从上述4种血清中,各挑选10份血清(阴性、阳性各5份)共40份,采用口蹄疫病毒液相阻断ELISA(LPB-ELISA)抗体检测试剂盒进行验证。结果显示:cELISA检测为阳性的20份血清中,用LPB-ELISA检出阳性19份;cELISA检测为阴性的20份血清中,用LPB-ELISA检出阴性17份;两种方法的κ值为0.8,总符合率为90.00%。结果表明,A型cELISA试剂盒与LPB-ELISA试剂盒的符合率和一致性均较高,可用于口蹄疫流行病学调查和血清学监测。     

湖南省部分地区病猪和屠宰猪群猪圆环病毒感染状况调查

为了解湖南省猪圆环病毒（PCV）流行情况,采用荧光PCR方法,对来自湖南省14个市州的821份临床病料和屠宰场猪淋巴结样品进行PCV1、PCV2、PCV3和PCV4荧光PCR检测。结果显示：在821份样品中,PCV阳性检出率为88.06%（723/821）,阳性检出率由高到低依次为PCV2（81.49%）、PCV3（33.98%）、PCV1（8.04%）和PCV4（1.10%）;278个样品存在两种及以上的PCV混合感染,以PCV2+PCV3混合感染最多,占82.01%;在地域分布上,PCV1、PCV2、PCV3在湖南省各地区均有分布,而PCV4分布范围较小,感染率低。结果表明,湖南省内PCV流行较为普遍,4种基因型病毒都存在,但以PCV2流行最为广泛,需要加强防控。     

新型莫西菌素浇泼剂对羊蜱蝇和消化道线虫驱虫药效评价

试验旨在通过新型莫西菌素（moxidectin,MOX）浇泼剂对羊的驱虫试验,评价MOX浇泼剂对羊蜱蝇和消化道线虫的临床药效。将150只阿勒泰羊经产母羊随机分为6组,每组25只,分别为不用药对照组、螨净药浴组、MOX注射组和MOX低（0.05 mL/（kg·BW））、中（0.1 mL/（kg·BW））和高（0.2 mL/（kg·BW））剂量浇泼组,试验期为1周。试验第1天按前述分组中方法和剂量驱虫1次。观察受试动物临床表现,测定给药前1天和给药后第7天体温、脉搏和呼吸（TPR）,血液生理、生化和尿常规指标;测定羊蜱蝇减虫率、治愈率。采取粪样测定消化道线虫每克粪中虫卵个数（EPG）减少率及转阴率。结果显示,与不用药对照组相比,给药后第7天各驱虫组TPR、血液生理、生化及尿常规指标均无显著差异（P>0.05）。螨净药浴组羊蜱蝇减虫率为43.7%,而MOX注射组及MOX低、中、高剂量浇泼组均为100%。螨净药浴组治愈率为13.0%,而MOX注射组及MOX低、中、高剂量浇泼组均为100%;与不用药对照组相比,螨净药浴组消化道线虫卵减少率为0,而MOX注射组及MOX低、中、高剂量浇泼组分别为91.6%、93.1%、94.9%和97.8%。螨净药浴组虫卵转阴率为0,而MOX注射组及MOX低、中、高剂量浇泼组分别为77.0%、69.2%、69.2%和84.6%。本试验结果表明,新型MOX浇泼剂对体外寄生虫和消化道线虫均有显著疗效,且优于螨净药浴,与MOX注射剂等效。实践应用中最适MOX浇泼剂量推荐为0.05 mL/（kg·BW）（即0.25 mg/（kg·BW））。     

哈萨克羊iNOS基因多态性与布鲁氏菌病的相关性分析

试验旨在探讨哈萨克羊诱导型一氧化氮合酶（iNOS）基因多态性与布鲁氏菌病的相关性。使用虎红平板凝集试验（RBPT）方法对231只哈萨克羊血清进行布鲁氏菌病血清学检测,参考GenBank中绵羊iNOS基因序列,针对其第6、7、8外显子及其邻近内含子片段设计引物,利用PCR-SSCP技术和DNA测序技术对231只哈萨克羊的iNOS基因进行多态性检测,分析其SNPs与哈萨克羊布鲁氏菌病易感性的相关性。结果表明,67只哈萨克羊为布鲁氏菌感染阳性,阳性检出率为29.00%。在哈萨克羊iNOS基因的外显子6和8片段上未检测到多态位点,在外显子7片段上检测出F7-T18054C和F7-C18084T 2个多态位点,在F7-T18054C多态位点上检测到3种基因型（TC、TT、CC）,优势等位基因和基因型分别是C型和CT型,其等位基因频率和基因型频率分别是0.660和0.446。在F7-C18084T多态位点上检测到2种基因型（CT、CC）,优势等位基因频率和基因型分别是C和CC型,其等位基因和基因型频率分别是0.946和0.892。F7-C18084T属于低度多态（PIC<0.25）,F7-T18054C属于中度多态（0.25 < PIC < 0.5）。相关性分析表明,F7-T18054C和F7-C18084T多态位点与布鲁氏菌病易感性无显著相关性（P>0.05）。试验结果表明,哈萨克羊iNOS基因F7-T18054C和F7-C18084T多态位点与布鲁氏菌病易感性不存在相关性。     

气肿疽梭菌CctA基因的原核表达及其间接ELISA检测方法的建立

研究旨在原核表达气肿疽梭菌细胞毒素A （CctA）基因,用纯化的重组蛋白建立其间接ELISA检测方法,a。利用大肠杆菌密码子的偏爱性优化CctA基因序列,克隆至原核表达载体pET-28a （+）,双酶切鉴定原核表达质粒pET28a-CctA并测序。将重组质粒转入大肠杆菌,经IPTG诱导得到高表达的重组CctA包涵体蛋白。包涵体蛋白变性后经镍（Ni）柱纯化,SDS-PAGE检测其纯化效果。以豚鼠抗气肿疽梭菌抗血清为一抗,用Western blotting方法检测重组CctA蛋白的反应原性。用棋盘滴定法建立间接ELISA检测方法,以复性后的重组CctA蛋白作为检测抗原,摸索抗原包被浓度、封闭液的种类及浓度、抗体的最适稀释度和反应条件等。选用3批气肿疽灭活疫苗免疫豚鼠后采集血清,同时用攻毒和间接ELISA两种方法验证疫苗的免疫效力。质粒双酶切结果显示,得到大小约853 bp的条带,与预期相符,且测序结果正确。SDS-PAGE结果表明,成功表达并纯化大小为35 ku的重组CctA蛋白。Western blotting结果显示,豚鼠抗气肿疽梭菌抗血清与重组CctA蛋白具有良好的反应原性。建立的间接ELISA法的最适条件为：抗原的包被浓度为0.5 μg/mL,于4 ℃包被过夜;封闭液选择10%胎牛血清,37 ℃孵育2 h;二抗的稀释度为1：8 000;室温避光显色10 min后终止反应,测定D_{450 nm}值。当P/N>4.6时,间接ELISA法检测结果与豚鼠攻毒试验结果拟合度较好。本研究成功表达CctA基因并纯化了重组CctA蛋白,建立的以重组CctA蛋白为检测抗原的间接ELISA检测方法,有望成为气肿疽灭活疫苗免疫效果验证的替代方法。