莫能菌素的诱变育种与高通量筛选

莫能菌素采用紫外线-氯化锂复合诱变育种,增加突变率,再用生测法进行初步筛选,以提高筛选效率。然后把两者结合运用,既提高筛选效率又提高了菌种的生产能力。     

羊同期发情腹腔镜输精最佳时间的研究

[目的]确定羊同期发情腹腔镜输精的最佳时间。[方法]选取繁殖机能正常、营养状况良好的适龄母羊,在试验母羊阴道内放置孕酮海绵栓10~14 d,撤栓时肌肉注射250~330 IU/只孕马血清促性腺激素（PMSG）。撤栓后12~24 h开始试情,早晚各1次。按撤栓后时间及发情时间不同分4组进行腹腔镜输精：1组为撤栓后46~50 h输精（n=138）,2组为撤栓后50~54 h输精（n=156）,3组为撤栓后54~58 h输精（n=275）,4组为羊发情后30~36 h输精（n=226）。输精后第40~45天进行B超检查,计算各组母羊受胎率。[结果]发情后30~36 h输精的母羊（4组）受胎率最高,为97.79%,其次为撤栓后50~54 h输精的母羊（2组）,受胎率为97.44%,撤栓后46~50 h输精的母羊（1组）受胎率为91.30%;撤栓后54~58 h输精的母羊（3组）受胎率最低,为89.82%。[结论]采用同期发情配合腹腔镜输精技术进行母羊人工授精,最佳输精时间为撤栓后50~54 h以及母羊发情后30~36 h。     

Multimarker and rare variants genomewide association studies for bone weight in Simmental cattle

Bone weight, defined as the total weight of the bones in all the forequarter and hindquarter joints, can reflect somebody conformation traits and skeletal diseases. To gain a better understanding of the genetic determinants of bone weight, we used a composite strategy including multimarker and rare‐marker association to perform genomewide association studies (GWAS) for that character in Simmental cattle. Our strategy consisted of three models: (i) A traditional linear mixed model (LMM) was applied (Q+K‐LMM); (ii) single nucleotide polymorphisms (SNPs) with p‐values less than .05 from the LMM were selected to undergo the least absolute shrinkage and selector operator (Lasso) in the second stage (LMM‐Lasso); (iii) genes containing two or more rare SNPs were examined by performing the sequence kernel association test (gene‐based SKAT). A total of 1,225 cattle were genotyped with an Illumina BovineHD BeadChip containing 770,000 SNPs. After the quality‐control procedures, 1,217 individuals with 608,696 common SNPs and 105,787 rare SNPs (with 0.001 < minor allele frequency [MAF] <0.05) remained in the sample for analysis. A traditional LMM successfully mapped three genes associated with bone weight, while LMM‐Lasso identified nine genes, which included all genes found by traditional LMM. Only a single gene, EPHB3, surpassed the significance threshold after Bonferroni correction in gene‐based SKAT. In conclusion, based on functional annotation and results from previous endeavours, we believe that LCORL, RIMS2, LAP3, PRKAR2B, CHSY1, MAP2K6 and EPHB3 are candidate genes for bone weight. In general, such a comprehensive strategy for GWAS may be useful for researchers seeking to probe the full genetic architecture underlying economic traits in livestock.     

基于GIMMS NDVI 3g.v1的近34年青海省植被生长季NDVI时空变化特征

在气候变化和人类活动的综合影响下,青海省生态环境发生了明显变化。在此背景下,以GIMMS NDVI 3g.v1为数据源,采用Sen+Mann-Kendal方法研究青海省1982-2015年植被覆盖区域NDVI时空变化,将趋势分析和R/S(rescaled range analysis)分析叠加,研究植被生长季NDVI变化的持续性特征,并揭示植被对气候变化及人类活动的响应规律。结果表明:1)近34年青海省植被NDVI整体呈从西北到东南的增加趋势;且变异系数显示,波动性较大地区集中在柴达木盆地周边和青南牧区西北部等植被NDVI较低的区域,波动性较小地区集中在祁连山东部、东部农业区和青南牧区东南部等植被NDVI较高的区域。2)近34年青海省植被NDVI整体呈增加趋势,增长率为0.38%·10a~(-1);且NDVI变化具有明显的阶段性,存在1994年和2000年两个突变点。3)近34年青海省植被改善区域(75.4%)远大于退化区域(24.6%),其中显著改善面积占植被覆盖区域面积的40.9%,退化区随时间变化在空间上表现出明显的转移现象。4)Hurst指数表明,青海省植被变化反持续性较强,趋势分析与Hurst指数叠加得出,由退化转为改善的区域占植被覆盖区面积的13.7%,由改善转为退化的区域占植被覆盖区面积的44.3%,另41.5%的区域无法确定未来变化趋势。5)青海省植被生长季NDVI受气候变化和人类活动的双重影响,且不同植被类型对气候变化的响应存在较大差异。     

Characterization of integrons-mediated antimicrobial resistance among Escherichia coli strains isolated from bovine mastitis

To assess the prevalence of antimicrobial resistance and class I integrons in Escherichia coli strains (n=58) isolated from bovine mastitis in Inner Mongolia, antimicrobial susceptibility and the presence of various types of integrons were characterized. Most isolates were susceptible to amikacin, colistin, ceftazidime, gentamicin and kanamycin, while those also exhibited high resistant incidence rates to ampicillin, amoxicillin, sulfadiazine and sulfamethoxydiazine. The integrase gene of integrons was amplified by PCR using degenerate primers. The integrons were confirmed by restriction fragment length polymorphism (RFLP) analysis of positive PCR products. Neither class II nor class III integron was detected, while 56.90% (n=33) of the isolates were positive for the presence of intI1 gene. Sequencing analysis of gene cassettes revealed that seven gene cassettes were found, which encoded resistance to trimethoprim (dfrA1 and dfrA17), aminoglycosides (aacA4, aadA1 and aadA5) and chloramphenicol (catB3), respectively. Of them, the gene cassette array dfrA17-aadA5 was found most prevalent (62.96%). The percentage of positive-integron among the isolates whose resistant profile was relatively broad (n> or =7) is 100.00%, while the one in narrow-profile isolates (n=2-6) is 30.56%. The correlation analysis revealed the incidence of integrons among the isolates were highly related to the resistant profile, indicating integrons play an important role in the dissemination and spread of the antimicrobial resistant strains.     

UPLC-MS／MS法同时测定鸡肉中的莫能菌素和盐霉素残留

建立了同时测定鸡肉中莫能菌素和盐霉素残留检测方法。在国内外已有研究的基础上,对样品前处理和仪器条件进行了改进,采用乙腈提取鸡肉中莫能菌素和盐霉素残留,石墨化碳黑SPE小柱对提取液进行净化,然后用Waters ACQUITY UPLCTMTQD超高效液相色谱-串联质谱仪测定。结果表明,莫能菌素和盐霉素的检测限为0.2μg/kg,最低定量限为0.5μg/kg。选取0.5、1.0、2.0μg/kg三个浓度进行空白添加回收率实验,回收率为71.9%~95.1%,批内变异系数和批间变异系数均小于15%,适合大量鸡肉样品中莫能菌素和盐霉素的残留检测。     

猪链球菌9型荚膜多糖cps9G基因的原核表达

本研究旨在克隆表达猪链球菌9型荚膜多糖部分抗原表位。设计1对引物,采用PCR法以猪链球菌9型河南分离株PY-2的基因组DNA为模板扩增出cps9G全基因序列。用限制性核酸内切酶BamHⅠ、XhoⅠ进行双酶切后,将其定向克隆到pET32a（＋）中,构建重组表达质粒pET32a—cps9G,并将其转化至大肠杆菌BL21（DE3）感受态细胞中进行诱导表达,并优化表达条件。结果表明,经1．2mmol／L IPTG诱导4h后SDS-PAGE分析表明,重组菌株表达出了约50700的融合蛋白条带,与预期一致。Western-blotting分析表明,该融合蛋白具有特异的生物学活性。这为以后建立快速、简便、特异的免疫学检测方法及制备单克隆抗体提供了较好的抗原,同时为研制猪链球菌亚单位疫苗和诊断试剂盒奠定了基础。     

模拟增温与施肥对高寒草甸土壤酶活性的影响

摘要：采用开顶式生长室模拟增温的方法,研究了气温升高和施肥对高寒草甸土壤酶活性的影响。研究表明,单独增温导致高寒草甸土壤纤维素酶和磷酸酶活性分别提高了12.4%、29.1%,而脲酶活性降低了18.0%,过氧化物酶和多酚氧化酶活性无明显变化,说明增温促进高寒草甸土壤中碳磷循环。在不增温条件下,施肥抑制了土壤纤维素酶、多酚氧化酶和过氧化物酶活性,而施NPK则提高了脲酶和磷酸酶活性。增温条件下,施肥不会引起土壤纤维素酶、多酚氧化酶和过氧化物酶活性的显著变化,而施可溶性碳肥使脲酶活性显著升高（P<0.05）,施NPK使磷酸酶活性显著降低。增温和施肥的交互作用对土壤纤维素酶、过氧化物酶、磷酸酶和脲酶活性有显著影响,而对多酚氧化酶无明显影响。因此,预测在未来气候变暖背景下,高寒草甸土壤中的多种酶活性对施肥的响应可能不显著。     

绵羊食管、网胃和瘤胃电活动的研究

用RM-6008型记录仪记录了绵羊在安静、进食、反刍和嗳气等状态下的食管、网胃和瘤胃的前背囊、后背盲囊、后腹盲囊、前腹囊的电活动及其先后次序。结果表明:食管电呈群集型,只在进食、嗳气和反刍时出现。在各种生理状态下,网胃电最先出现,波型随生理状态不同而有差异。瘤胃电由第一次波(P波)和第二次波(S波)组成;P波开始于网胃电活动之后,出现频率与网胃电频率相同,其活动次序为,前背囊→后背囊→后腹盲囊→前腹囊,有时电活动止于后背盲囊;S波位于P波之后,其频率低于P波,且与进食和嗳气有关;进食时,S波出现频率最高,其活动次序是,前腹囊→后腹盲囊→后背盲囊→前背盲囊;嗳气时,S波活动次序是,后腹盲囊→后背盲囊→前背囊;安静和反刍时的活动次序与嗳气时相似。     

氨化油菜秸秆对威宁黄牛饲养效果的影响

为了提高秸秆利用率,试验将油菜秸秆通过氨化处理饲喂威宁黄牛,研究秸秆氨化前后营养成分及威宁黄牛生产性能的变化,并进行了经济效益分析。结果表明:与对照组相比,试验组日增重显著提高(试验组日增重为0.59 kg,对照组日增重为0.48 kg)(P0.05),料重比显著降低(试验组料重比为14.42,对照组料重比为17.17)(P0.05),试验组效益增加1.89元/(d·头)。说明氨化油菜秸秆替代30%粗饲料能提高威宁黄牛日增重,降低料重比,带来较好的经济效益。