透明颤菌血红蛋白基因(vgb)在大肠杆菌中的高效表达

利用PCR技术将透明颠菌血红蛋白基因（vgb）克隆到融合表达载体pET28a，在大肠杆菌BL21（DE3）表达。重组蛋白在30℃诱导获得可溶表达。利用Ni^2＋亲和层析对重组蛋白进行了纯化，得到重组的透明颠菌血红蛋白，蛋白呈红色。实验现象显示重组蛋白与血红索相结合。紫外光区和可见光区波长扫描分析显示：蛋白粗提物和纯化后的血红蛋白在230nm和413nm都有强吸收峰，初步表明，尽管重组蛋白比天然蛋白多36个氨基酸，重组蛋白仍然具有生理活性。诱导后4h和6h的培养液氨基乙酰丙酸含量分别达到17mg／L和21mg／L。说明重组蛋白的表达明显促进了体内heme合成途径。     

鸡蛋消毒方法的对比研究

采用5种消毒方法对鸡蛋蛋壳表面细菌进行杀菌效果试验,结果表明,各处理组之间抑菌效果差异显著(P＜0.05),杀菌效果优劣依次为开水浸烫＞甲醛熏蒸＞过氧乙酸熏蒸＞高锰酸钾浸泡＞过氧乙酸浸泡,开水浸烫操作技术难度较大,容易引起蛋壳破裂,适宜实际生产的消毒方法为过氧乙酸熏蒸和甲醛熏蒸.将消毒处理后的鸡蛋在温度25℃,相对湿度60%～80%贮存20天,开水浸烫处理组鸡蛋仍保持较鲜的品质,其余各组鸡蛋品质较差.结果表明,常规消毒方法(除开水浸烫法)只能抑制蛋壳表面微生物的生长繁殖,不能延缓鸡蛋品质下降达到保鲜鸡蛋的作用.     

Virulence traits of avian pathogenic (APEC) and fecal (AFEC) <Emphasis Type="Italic">E. coli</Emphasis> isolated from broiler chickens in Algeria

Avian pathogenic E. coli (APEC) is the etiologic agent of avian colibacillosis, the most common disease responsible for chicken morbidity in the world. Although multiple virulence-associated factors were identified, their prevalence in Algeria is still poorly known. In the present research, 92 avian pathogenic E. coli (APEC) isolates were recovered from broilers with clinical signs and lesions of colibacillosis. In addition, 32 E. coli isolates collected from feces of healthy birds (AFEC) were included for comparison. All isolates were investigated by PCR for the presence of a total of 11 virulence-associated genes described for avian pathogenic (iroN, ompT, hlyF, iss, iutA, and fimC) and diarrheagenic E. coli (eae, stx, elt/est, ipaH, and aggR). The sensitivity of 39 APEC isolates to 16 antibiotics was also determined using antimicrobial pretreated microplates. Here, we report that 98% of the examined isolates host at least one of the tested virulence factors. The most prevalent genes in APEC were iutA (90.6%), ompT (86.9%), and iss (85.8%); whereas, iutA (78.1%), fimC (78.1%), and iroN (68.7%) were the highest prevalent genes in AFEC. Our data showed that none of the AFEC isolates harbor any of the tested diarrheagenic genes. Moreover, only elt/est (5.4%), stx (2.1%), and ipaH (2.1%) genes were carried by APEC isolates. We further established that ceftazodime, ceftiofur, mequindox, amoxicillin/clavulanic acid, and meropenem were the most efficient antibiotics against the analyzed APEC isolates. Overall, our findings provide more insights about APEC and AFEC virulence potential in Algeria which could participate in the fight against colibacillosis.     

Effects of in ovo feeding of L‐arginine on the development of digestive organs,intestinal function and post‐hatch performance of broiler embryos and hatchlings

This study was to investigate the effects of in ovo feeding (IOF) L‐arginine (Arg) solution on the development of digestive organs, the duodenal mucosa of broiler embryos and hatchlings, and the growth performance of chicks during the first week post‐hatch. A total of 720 fertilized eggs with similar weight were randomly allocated to three groups, consisting of eight replicates of 30 eggs each. Three treatments were arranged as non‐injected control, diluent‐injected (0.75% NaCl solution) group and Arg solution‐injected group containing 1% Arg, dissolved in diluent. At 17.5 days of incubation, 0.6 ml of IOF solution was injected into amniotic fluid of each egg of injected groups. Results showed IOF of Arg solution increased (p < .05) the chick embryo weight at 19 days of incubation; the body weight gain of post‐hatch broilers during 1–7 days; the weights of liver, pancreas, proventriculus and gizzard; the concentrations of duodenal ghrelin, vasoactive intestinal peptide and glucagon‐like peptide 2; and the duodenum mucosal enzyme activities of alkaline phosphatase, maltase, sucrase and inducible nitric oxide synthase of 7‐day‐old post‐hatch broilers compared with other groups. The IOF of Arg solution also increased (p < .05) the villus height (VH) and the ratio of VH to crypt depth (CD) and decreased (p < .05) the CD in duodenum of broiler embryos and post‐hatch hatchlings, except for the CD at 19 days of incubation. In conclusion, IOF of 1% Arg solution into the amnion at 17.5 days of incubation could improve the development of digestive organs, the duodenal morphology, the releasing of gastrointestinal hormones and mucosal enzyme activities of broiler embryos and hatchlings and finally the growth performance of chicks during the first week post‐hatch. Therefore, IOF of appropriate Arg solution could be an effective technology for regulating early nutrition supply and subsequent growth development in poultry industry.     

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.     

气候变化对通辽草甸草原草本植物物候期的影响

植物物候期的变化直接反映气候变化,并成为全球变化研究的重点。基于通辽市巴雅尔吐胡硕牧业气象试验站1981-2016年主要植物羊草(Leymus chinensis)、冰草(Agropyron cristatum)、委陵菜(Potentilla chinensis)、车前(Plantago asiatica)物候观测数据和同期气象观测数据,研究了气候变化对草本植物返青期、黄枯期和生长季的影响,结果表明:1)研究区近36年来年均气温上升0.35℃·10 a-1(P0.01),年均降水量减少22.73 mm·10 a-1,气候暖干化特征明显。2)草本植物返青期呈一致性延迟趋势,平均每10年延迟1.16~7.60 d。羊草、车前返青推迟主要受2-3月累计降水量减少的影响,冰草、委陵菜返青推迟主要受2-4月、3-4月累计降水量减少的影响。3)草本植物黄枯期变化趋势不同,羊草、冰草黄枯期延迟主要受夏季、初秋累计降水量影响,委陵菜黄枯期提前主要受夏季、初秋的累计降水量和日照时数共同影响,车前草黄枯期延迟主要受夏季、初秋平均气温影响。4)草本植物生长季长度主要受气温和降水量影响。气候变暖后,研究区夏季、生长季平均气温显著上升,降水量逐渐减少,导致羊草、冰草、委陵菜生长季缩短;而车前对气温反应比较敏感,气温升高有利于车前生长季延长。     

旱、盐胁迫下黄芪种子萌发及其对水杨酸的响应

通过测定不同浓度聚乙二醇(PEG)和NaCl胁迫下膜荚黄芪(Astragalus membranaceus)和蒙古黄芪(A.membranaceus var.mongholicus)种子的最终发芽率、发芽势、简化活力指数、苗长、根长、苗干重、根干重等指标,以及不同施用方式和浓度的水杨酸(SA)对重度PEG和NaCl胁迫下两种黄芪种子最终发芽率的影响,旨在研究两种黄芪种子对PEG和NaCl胁迫响应方式的异同及SA对两种黄芪种子在重度PEG和NaCl胁迫下保护效果的差异。结果表明,低浓度PEG和NaCl只能促进膜荚黄芪种子萌发。在-0.5 MPa PEG和-0.7 MPa NaCl处理下,蒙古黄芪种子的活力指数显著优于膜荚黄芪种子(P0.05)。SA浸种能促进重度PEG和NaCl胁迫下两种黄芪种子的最终发芽率。蒙古黄芪种子的耐胁迫性高于膜荚黄芪种子。两种黄芪种子对SA的响应方式有区别。SA浸种处理优于拌种处理。     

黄花苜蓿花药培养再生体系的建立与单倍体鉴定

花药组织培养再生体系的构建是单倍体育种的重要途径之一。本研究对呼伦贝尔黄花苜蓿(Medicago falcate‘Hulunbeier’)进行了花药组织培养研究并建立了花药组培再生体系。结果显示,液体悬浮培养基比固体培养基更适合呼伦贝尔黄花苜蓿花药愈伤组织的培养,其愈伤形成的培养条件为B5培养基+0.5 mg·L~(-1) 2,4-D+0.25 mg·L~(-1) 6-BA+0.4mg·L~(-1) NAA+3.0mg·L~(-1) KT;分化培养基为MS+1.0mg·L~(-1) 2,4-D+0.5mg·L~(-1) 6-BA+2%蔗糖+0.7%琼脂;生根培养基为1/2MS+0.1mg·L~(-1) NAA+2%蔗糖+0.7%琼脂;获得的再生植株经流式细胞仪鉴定,其单倍体比例高达27%。     

过量表达甘菊CBF1基因提高拟南芥抗旱耐盐能力

CBF是一类植物特异性、具有多种功能的转录因子。本研究将从甘菊(Chrysanthemum lavandulifolium)中分离的ClCBF1基因构建到植物表达载体pBI121上,并采用农杆菌介导的花序浸染法对拟南芥(Arabidopsis thaliana)进行遗传转化,共获得4个转基因株系经过抗性筛选和RT-PCR验证,对其中表达量较高的3个转基因株系进行研究,结果表明:在150mmol·L~(-1)甘露醇培养基中转基因拟南芥的种子萌发率和根长平均为野生型的2.0倍和1.2倍,在150mmol·L~(-1) NaCl培养基中转基因拟南芥种子萌发率和根长平均为野生型的1.1倍和1.4倍;在干旱和盐胁迫条件下,转基因拟南芥幼苗的成活率高于野生型,并且超氧化物歧化酶(SOD)和过氧化物酶(POD)活性显著高于野生型(P0.05),丙二醛(MDA)含量及相对电导率显著低于野生型植株(P0.05)。说明ClCBF1基因在拟南芥受到干旱和盐胁迫过程中发挥一定作用。     

陕西省榆林市某鸡场家禽H7N9流感紧急流行病学调查

2017年5月21日,陕西省榆林市某养殖场的蛋鸡出现异常死亡。经国家参考实验室确诊,该起疫情为H7N9流感疫情。为找出引发疫情的可能原因,采用描述性流行病学方法,描述疫情的三间分布,分析发病的风险因素。调查结果显示:该场饲养规模较大,共有8栋鸡舍,饲养4.5万只蛋鸡;疫情暴发突然,病程仅为5 d;蛋鸡死亡2.2万只,死亡率为48.9%;疫情具有明显的场内传播规律,从靠近鸡场入口的鸡舍逐渐向内蔓延;产蛋高峰期蛋鸡的发病风险是雏鸡的37.458倍,差异显著(P0.05);所分离毒株的HA基因裂解位点有4个氨基酸插入,为变异株;该起疫情可能是通过运输淘汰蛋鸡的车辆、野鸟、饲料和饮水,以及调入雏鸡等途径引入的。疫情发生后,当地政府按照《高致病性禽流感防治技术规范》,发布了封锁令,划定了疫点、疫区和受威胁区;在疫区采取了扑杀、无害化处理、关闭疫区内所有禽类产品交易市场等控制措施。采取以上处置措施后,疫情得以快速有效控制。