转露地菊CgDREB22基因的烟草抗逆性分析

DREB转录因子是AP2/ERF转录因子的一个亚家族,主要参与植物对干旱、高盐和低温等逆境的分子应答机制。前期研究从露地菊中分离出一个CgDREB22基因,构建植物过表达载体并用农杆菌介导法转化烟草。对获得的T₁代转基因烟草种子和幼苗进行高盐(150 mmol·L^-1 NaCl )、干旱(150 mmol·L^-1甘露醇)、低温(4 ℃)等处理,进而观察表型并测定相关生理指标。结果表明:在低温、干旱和高盐胁迫下,转基因烟草幼苗鲜重和根长都低于野生型;在高盐和干旱胁迫下,转基因烟草幼苗脯氨酸含量、抗氧化物酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)]活性均显著低于野生型。综上说明,露地菊CgDREB22基因在植物受到非生物胁迫的过程中起负调控的作用。     

多胎基因FecB对德×寒×细三元杂交羊产羔性能的影响

为了研究FecB基因在德×寒×细三元杂交羊群体中的多态性分布及与产羔性能指标的相关性,该试验将以德国肉用美利奴羊（以下简称德肉美）为父本,以小尾寒羊公羊与东北细毛羊母羊级进杂交生产的F₂代羊为母本（以下简称寒×细F₂母羊）,生产的三元杂交后代羊（以下简称德×寒×细三元杂交羊）作为研究对象,采用KASP方法分析FecB基因型并计算基因频率和基因型频率,分析FecB基因频率与德×寒×细三元杂交羊群体产羔性能的相关性。结果发现,德肉美并不含有FecB基因,德×寒×细三元杂交羊B等位基因频率增加明显且处于Hardy-weinberg不平衡状态,提示人工选择对杂交羊群体FecB基因富集影响较大。对寒×细F₂代母羊和德×寒×细三元杂交羊群体不同基因型个体产羔数进行秩和检验分析发现,P值均小于0.01,平均产羔率均表现为++基因型小于B+和BB基因型（P<0.05）,说明B等位基因对德×寒×细三元杂交羊群体产羔性能的提升发挥重要作用。该试验为利用FecB基因提升德×寒×细三元杂交羊群体产羔性能提供参考。     

“杜蒙萨”三元杂交羔羊产肉性能测定试验

为更好地指导乌兰察布市优质育肥羔羊生产体系建设,进一步了解在相同的饲养管理条件下,“杜蒙萨”三元杂交羔羊与当地蒙古羊羔羊在养殖效益方面的对比情况,进行了“杜蒙萨”三元杂交羔羊产肉性能试验。试验分为试验组（“杜蒙萨”三元杂交羔羊组）和对照组（蒙古羊羔羊组）2个组,每组采取随机取样方式选择试验用羊。经过体重测定与屠宰试验测定,统计试验数据,通过分析试验数据进行经济效益对比。试验结果显示,在自然环境和饲养管理条件完全相同的条件下,经过对生长及产肉性能测定数据的整理、统计和分析,“杜蒙萨”三元杂交组羔羊平均初生重、断乳重、5月龄出栏活重、5月龄平均日增重均高于对照组蒙古羊羔羊。试验充分证明了在牧区推广“杜蒙萨”三元杂交羊生产体系建设,是实现牧区草原生态高效养殖和促进农牧民养殖户提质增效的有效途径之一。     

柱花草SgNCED1基因的克隆及功能分析

脱落酸(ABA)作为一种调控生长发育的激素,对植物多种生理过程起着调控作用。9-顺式环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase,NCED)是高等植物ABA生物合成途径的关键酶。NCED基因受逆境胁迫诱导,导致逆境ABA的积累。研究NCED基因克隆、表达,调控ABA的生物合成,是提高植物抗逆性的重要途径。柱花草(Stylosanthesgu ianensis)是一种热带和亚热带地区广泛种植的耐旱草种。     

不同地区种植的豫玉22营养成分分析

本文对河南6个地区生产的豫玉22的常规营养成分(粗蛋白质、粗脂肪、粗灰分、水分、钙、磷)和微量元素(铁、铜、锰、锌)进行了测定。结果表明:常规营养成分和微量元素因种植地区的不同而存在明显差异。研究结果将为饲料企业选择饲料原料提供理论依据。     

光敏型高丹草在陇东旱塬的生物学特性和营养成分比较研究

在陇东旱塬地区研究了海牛、BJ0603和大卡,共3个光敏型高丹草品种的生物学特性、不同生育时期营养成分变化及刈割次数对产量的影响。结果表明:4月下旬播种、10月上旬收获时,各品种都处于抽穗或开花期,不能完成生殖生长。收获前测定表明,品种间在株高、节间数、叶数和茎粗方面差异不显著(P>0.05),但叶片和单秆重差异显著(P<0.05);每个品种全年刈割1茬的标准草产量(65%含水量)和干物质产量均高于全年刈割2茬的,但差异不显著(P>0.05),全年刈割2茬的鲜草产量极显著高于(P<0.01)1茬的。无论全年刈割1茬或2茬,品种间的鲜草、标准草(65%含水量)和干物质产量均差异显著(P<0.05),产量依次为海牛>BJ0603>大卡;从拔节前期到孕穗期,各品种茎秆中粗蛋白(CP)、粗脂肪(EE)含量呈下降趋势;酸性洗涤纤维(ADF)、中性洗涤纤维(NDF)含量呈上升趋势,粗纤维(CF)、可溶性糖含量呈先升后降的趋势;粗灰分(ASH)含量呈直线下降的趋势;无氮浸出物(NFE)含量呈先下降后上升的趋势。茎秆中CP、ADF、NDF、ASH、NFE及可溶性糖含量在品种间差异显著(P<0.05),但EE、CF含量差异不显著(P>0.05)。综合考虑产量和养分含量变化,我们认为,在陇东旱塬地区春播光敏型极晚熟高丹草生产模式中,作为青贮利用时,以全年刈割1茬且在霜降前收割最佳;作为干草调制或青饲利用时,以全年刈割2茬且第1茬在拔节后期孕穗前刈割,第2茬在霜降前刈割最佳。     

流产型布鲁氏菌Omp22基因酵母双杂交诱饵载体的构建及鉴定

通过PCR获得流产型布鲁氏菌Omp22基因的编码区,将其克隆到pSOS载体中,构建酵母双杂交系统的诱饵载体pSOS-Omp22。测序正确后,将重组质粒导入cdc25H检测其表达产物对酵母细胞有无毒性和自激活作用,且在酵母细胞中定位是否正确。结果表明,获得了正确的流产型布鲁氏菌Omp22基因编码区,并成功克隆到pSOS诱饵载体中,且转化有诱饵载体的cdc25H在SD/Glucose(-L)营养缺陷平板上生长良好,说明表达产物对酵母细胞无毒性,对报告基因也无自激活作用且其定位正确。表明pSOS-Omp22可应用在酵母双杂交系统中,用于寻找巨噬细胞cDNA文库中与Omp22相互作用的蛋白质。     

Quantitative Analysis of Stress Echocardiograms in Healthy Horses with 2‐Dimensional (2D) Echocardiography,Anatomical M‐Mode,Tissue Doppler Imaging,and 2D Speckle Tracking

Background: Stress echocardiography is used to diagnose myocardial dysfunction in horses, but current methods are not well standardized. The influence of heart rate (HR) on measurements is largely unknown. Objectives: To investigate the use of 2‐dimensional echocardiography (2DE), anatomical M‐mode (AMM), tissue Doppler imaging (TDI), and 2D speckle tracking (2DST) at rest and after exercise for quantification of regional and global left‐ventricular (LV) function. Animals: Five athletic Warmblood horses; 11.6 ± 3.6 years; 529 ± 48 kg. Methods: Prospective study. Three separate echocardiographic examinations were performed before (baseline) and over 5 minutes after treadmill exercise with 2DE (1st, short‐axis view; 2nd, long‐axis view) and pulsed‐wave TDI (3rd examination). Offline analyses were performed at baseline and after exercise at HR 120, 110, 100, 90, and 80 minute^?1. Global and segmental measurements were compared by analysis of variance. Results: Quantitative analyses of stress echocardiograms were feasible in all horses. None of the AMM indices changed significantly after exercise. Stroke volume and ejection fraction by 2DE and strain by 2DST decreased, whereas strain rate by 2DST increased significantly at HR > 100 minute^?1. TDI analyses were technically difficult and provided little additional information. Conclusions and Clinical Importance: Volumetric indices by 2DE and strain and strain rate by 2DST are applicable for quantitative assessment of stress echocardiograms. In healthy horses, they are significantly altered at a HR > 100 minute^?1 and need to be evaluated in view of the instantaneous HR. Further investigations are needed to define the clinical value of stress echocardiography in horses with cardiac disease.     

Efficiency of microbial phytase supplementation in diets formulated with different calcium:phosphorus ratios,supplied to broilers from 22 to 33 days old

An experiment was conducted with broilers from 22 to 33 days of age to evaluate the efficiency of six microbial phytases supplemented in diets (1500 FTU/kg) that were formulated with three different calcium:available phosphorus (Ca:P_avail) ratios (4.5:1.0, 6.0:1.0 and 7.5:1.0). A positive control diet without phytase was formulated with a Ca:P_avail ratio of 7.5:3.4 to meet the nutritional requirements of the broilers. The P and ash contents of the tibia, magnesium in the plasma, performance, balance and retention of phytate phosphorus (P_phyt), intake of total P and nitrogen (N), nitrogen‐corrected apparent metabolizable energy and apparent digestibility of dry matter of the diets were not influenced (p > 0.05) by the type of phytase or the dietary Ca:P_avail ratio. However, there was an interaction (p < 0.05) between the phytase type and the Ca:P_avail ratio for the retention coefficients of total P, Ca and N. Phytase B resulted in the highest Ca deposition in the tibia (p < 0.01). Phytases D, E and F reduced the Ca concentrations in the tibia (p < 0.01) and plasma (p < 0.05). Phytase D increased the P level in the plasma and decreased the total P excretion (p < 0.01). Phytases E and F increased Ca excretion, while phytase A reduced it (p < 0.01). Regardless of the phytase type, increasing the dietary Ca:P_avail ratio reduced (p < 0.05) the plasma P concentration and the excretion of total P and N and, conversely, increased (p < 0.05) the plasma concentration, intake and excretion of Ca. For the rearing period evaluated, it is possible to reduce the P_avail of the diet to 1.0 g/kg when Ca is maintained at 7.5 g/kg, and the diet is supplemented with 1500 FTU of phytase A, C, D or E/kg. This diet allows the maintenance of performance and adequate bone mineralization, and it improves the Ca, total P and P_phyt utilization in addition to reducing the excretion of N and P into the environment.     

Analysis of results from 21 years of milking system inspections in Japanese dairy farms

To prevent mastitis caused by inappropriate milking systems, inspection of the system, and maintenance of optimal function are crucial. This study aimed to clarify the problems with milking systems in Japan by analyzing the results of milking system inspections over the past 21 years. A total of 190 dairy farms (358 systems; 153 high‐line systems, 205 low‐line systems) were inspected for the checkpoints of problems originating from installation (PI) or problems originating from insufficient maintenance (PIM). Results were divided into initial and periodic inspections, then analyzed by year of inspection or years elapsed since equipment installation. With increasing years, inadequacy of milk piping and regulator maintenance tended to increase for high‐line systems. On the other hand, defects in milking units tended to increase for low‐line systems. This difference was attributed to the structures of these milking systems. The present study revealed potential problem areas in high‐ and low‐line milking systems from recent years. To maintain normal functioning in milking systems, the frequency and content of inspections as currently being conducted need to be reviewed.     

Genetic Analysis of Dobrava–Belgrade Virus from Western Serbia – A Newly Detected Focus in the Balkan Peninsula

Dobrava–Belgrade virus (DOBV) is a hantavirus species that causes the most severe form of haemorrhagic fever with renal syndrome (HFRS) in Europe. DOBV has been detected in three Apodemus rodents: A. flavicollis, A. agrarius and A. ponticus. These emerging viruses appear throughout the Balkan Peninsula including Serbia as its central part. In this study, we examined the seroprevalence, molecular epidemiology and phylogenetics of DOBV from A. flavicollis captured at six Serbian localities. Furthermore, we applied microsatellite typing of host animal genome to analyse the role of host kinship in DOBV animal transmission. The overall IgG seropositivity rate over 3 years (2008–2010) was 11.9% (22/185). All seropositive samples were subjected to RT‐PCR and DNA sequencing for S and L genome segments (pos. 291–1079 nt and 2999–3316 nt, respectively). DOBV was genetically detected in three samples from mountain Tara in western Serbia, a newly detected DOBV focus in the Balkans. No sequence data from human cases from Serbia are available for the studied period. However, collected DOBV isolates in this work phylogenetically clustered together with isolates from Serbian human cases dating from 2002, with 1.9% nucleotide divergence. We determined the level of kinship between seropositive and seronegative animal groups and found no significant difference, suggesting that horizontal virus transmission in the studied population was the same within and among the hatches. Our findings are the first genetic detection of DOBV in rodents in Serbia. We confirm wide and continuous hantavirus presence in the examined parts of the Balkans, underlying the necessity of continual monitoring of hantavirus circulation in A. flavicollis.     

贵州省猪源沙门氏菌对β-内酰胺类药耐药性及耐药基因分析

为了解贵州省猪源沙门氏菌对β-内酰胺类抗菌药物耐药性及其耐药基因的流行情况,本试验从贵州省9个地区规模养猪场中分离鉴定130株沙门氏菌,采用微量肉汤稀释法测定其对常用的8种β-内酰胺类抗菌药物的敏感性,并用PCR法对β-内酰胺酶耐药基因进行检测。结果显示,沙门氏菌对常用的β-内酰胺类抗菌药物耐药性十分严重,其中对头孢他啶的耐药率为100%,其次是氨苄西林和阿莫西林,耐药率分别为80.77%和76.15%,耐药率最低的是头孢噻呋和头孢氨苄,均为46.15%。所有菌株均为多重耐药,其中最少为二重,占总数的2.31%,最多为八重,占总数的4.62%,多重耐药主要集中在四至七重,占总数的88.46%。PCR结果显示,SHV耐药基因未检出,TEM、OXA、CTX-M 3种基因检出率分别是85%、75%和46%,细菌的耐药性与相关耐药基因的检出率基本呈正相关。结果表明,猪源沙门氏菌对β-内酰胺类药物具有普遍耐药性,其中头孢他啶尤为严重。TEM、OXA、CTX-M基因是贵州省猪源沙门氏菌主要耐药基因,临床日益严重的耐药现象与耐药基因的普遍存在有很大的关系。     

Molecular and Statistical Analysis of Campylobacter spp. and Antimicrobial‐Resistant Campylobacter Carriage in Wildlife and Livestock from Ontario Farms

The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial‐resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross‐sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial‐resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter‐specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C.  coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between livestock and wildlife samples, and the lack of similarity in molecular subtypes and AMR patterns, we concluded that the sharing of Campylobacter species between livestock and mammalian wildlife was uncommon.