牛轮状病毒RT-LAMP检测方法的建立

建立一种适用于牛轮状病毒(BRV)的反转录-环介导等温扩增(RT-LAMP)的快速检测方法。根据GenBank中登录的BRV VP7基因序列,针对其保守区设计了4条引物,利用RT-LAMP方法进行检测,并优化了反应体系与反应时间,检测了该方法的特异性和灵敏度。结果表明,该方法在等温条件下只需要50min就能检测出结果,与普通RT-PCR相比,具有较短的检测时间、良好的特异性和较高的灵敏度。论文针对BRV建立的RT-LAMP快速检测方法操作简便、无需昂贵设备、结果可视,适用于BRV在基层的快速检测。     

基于牛呼吸道合胞体病毒F基因TB Green Ⅱ荧光定量PCR检测方法的建立

为了对牛呼吸道合胞体病毒(bovine respiratory syncytial virus, BRSV)进行准确定量检测，针对BRSV F基因设计特异性引物，构建pMD19T-BRSV-gF重组质粒，以其为模板优化反应条件及反应体系，建立TB GreenⅡ荧光定量PCR方法并应用于临床BRSV检测。结果显示，针对BRSV F基因建立的TB GreenⅡ荧光定量PCR方法标准曲线线性关系良好，相关系数为0.995 2;灵敏度高，BRSV最低检测限达3.9×10¹ copies/μL;特异性良好，可特异性检测出BRSV,对牛传染性鼻气管炎病毒和牛病毒性腹泻病毒的扩增呈阴性；重复性好，批内和批间重复性试验变异系数均小于2%。对50份临床样品进行检测，荧光定量PCR对BRSV的阳性检出率(26%)明显优于常规PCR方法(14%)。结果表明针对BRSV F基因建立的TB GreenⅡ荧光定量PCR检测方法适合BRSV的临床检测。     

猪繁殖与呼吸综合征病毒RT-LAMP检测方法的建立

针对猪繁殖与呼吸综合征病毒(PRRSV)ORF7基因设计了2对特异性引物,建立了快速检测猪繁殖与呼吸综合征病毒的环介导等温扩增(RT-LAMP)方法,并进行了酶切鉴定以及特异性、灵敏性和临床试验确定。酶切鉴定试验结果与RT-LAMP结果完全符合;特异性试验结果表明本研究所建立的RT-LAMP检测方法只针对猪繁殖与呼吸综合征病毒;灵敏度试验结果显示RT-LAMP是RT-PCR方法的100倍;临床试验中RT-LAMP方法的检出率与病毒分离的符合率为100%。本检测方法操作简单,不需要复杂的仪器,是适合基层和现场检测而无需送至诊断实验室的快速灵敏实用的分子生物学检测方法。     

羊布鲁氏菌病的实验室常用检测方法对比

对于羊布鲁氏菌病的实验室检测,比较常用的方法包括虎红平板凝集试验(RBPT)、试管凝集试验(SAT)、补体结合试验(CFT)以及酶联免疫吸附试验(ELISA)等,这几种方法各有优势。笔者主要针对前两种检测方法,对比其检测的敏感性和准确性,希望能够对防控布鲁氏菌病起到一定的积极作用。     

几种布鲁氏菌病检测方法的比较

为研究虎红平板凝集试验(RBT)、全乳环状试验(MRT)、间接酶联免疫吸附试验(iELISA)及胶体金检测试纸条对布病的检测效果,本研究采用以上方法分别对感染地区和净化地区的牛群血清、羊群血清、牛群奶样进行了检测效果的比对。结果显示,针对牛群血清,RBT结果阳性率最高,iELISA与胶体金结果阳性率相似,表明RBT的敏感性较好,iELISA与胶体金检测试纸条的特异性较RBT好;针对羊群血清,iELISA与胶体金检测试纸条的敏感性和特异性较RBT好;针对牛群奶样,MRT结果阳性率低于全乳iELISA和胶体金检测试纸条。全乳iELISA和胶体金检测试纸条在试验中显示出具有较高的敏感性和特异性。     

鸡传染性支气管炎病毒RT-LAMP快速检测方法的初步建立

鸡传染性支气管炎(Avian Infectious Bronchitis,IB)是由鸡传染性支气管炎病毒(Infectious Bronchitis Virus,IBV)引起的一种急性、高度接触传染性呼吸道疾病.该病造成了巨大的经济损失,使感染的肉鸡被淘汰,产蛋鸡产量下降. 目前针对IBV的检测方法主要有病原分离、RT-PCR、IBV Real-time PCR检测方法、基因芯片检测方法等,以上方法虽然也可达到检测结果准备、敏感、特异高的特点,但是操作过程复杂且需要较为昂贵的仪器,不适合在基层推广使用.针对上述方法的缺陷,本试验旨在建立一种快速、简便、经济实用的IBV检测方法.     

基于部位组合HOG特征的行人检测

本文针对传统的基于梯度直方图特征(HOG)的行人检测耗时长的问题,提出结合头部与腿部特征的行人检测方法,根据腿部和头部的检测结果,综合采用部位约束,特征转化以及分类器阈值调整的方法对检测结果融合,大幅提升对于行人的辨识能力。     

鲤浮肿病毒环介导等温扩增(LAMP)检测方法的建立及应用

为了给鲤浮肿病(carp edema virus disease,CEVD)的防控提供特异、灵敏、快速的检测方法,针对鲤浮肿病毒(carp edema virus,CEV)的P4a基因序列设计5条特异性引物,建立CEV的环介导等温扩增检测方法(loop-mediated isothermal amplification,LAMP)。结果表明该方法具有很高的特异性。最低可检出6 000拷贝/μL,检测灵敏度与Real-time PCR方法相当。对30个鲤及锦鲤养殖场样品进行检测并与Real-time PCR方法进行比较,该方法的检测结果符合率达到96.7%以上。本方法的建立及应用为CEVD的防控提供依据。     

反转录环介导等温扩增技术快速检测猪水疱病毒方法的建立

为了建立一种针对猪水泡病毒(SVDV)的反转录环介导等温扩增技术(RT-LAMP),用于减少猪水泡病与口蹄疫因临床症状相似而造成的误诊,试验针对SVDV VP1基因序列设计4个LAMP引物,其中外侧引物用于SVDV PCR检测,并利用RT-LAMP和RT-PCR方法进行敏感性试验、特异性试验以及临床样品检测。结果表明:针对SVDV的RT-LAMP检测方法的敏感性是RT-PCR的10倍; RT-LAMP方法的特异性良好,与口蹄疫病毒无交叉反应;两种方法对临床样品检测的符合率为100%。说明试验建立的RT-LAMP方法具有敏感性高、特异性强、快速等特点,适合实验室和临床对SVDV的快速诊断。     

猪瘟病毒NASBA-RT-LAMP快速检测方法的建立

本研究基于核酸序列依赖性扩增技术(NASBA)和反转录环介导等温扩增技术(RT-LAMP),建立了一种针对猪瘟病毒的,快速、灵敏、特异的两步法检测技术。该方法的检测灵敏度与反转录巢式PCR(RT-NestPCR)相当,最低能检测出46 pg/μL的病毒RNA。该方法的建立为猪瘟病毒的快速诊断提供了新思路。     

三探草坪的起源、演化及其概念——几种外文、外语“草坪”一词及其他

初步发掘、整理了英、德、拉丁、俄、日等五种文字、语言中草坪一词内积淀的有关草坪起源与演化的信息。三证草坪草的分化 ,草坪的起源与早期演化均属自然。人类活动于草坪中 ,认识、利用和发展了草坪 ,可以劲直、也可以曲折。人类利用草坪的方式 ,无论中、外大致相同 ,利用的水平与社会整体发展的水平大体相应。草坪由草地分化发育 ,是草地的一个特殊类型。     

2017年国内外家兔遗传育种研究进展

作者对2017年国内外家兔遗传育种（传统育种、分子育种、繁殖技术）相关的研究进展进行了综述，发现国内外研究侧重不同，且与往年相比存在明显变化。国内传统育种主要包括品种选育和性状评定两方面研究，并取得了良好的进展；在分子育种领域做了较多的研究，主要涉及性状包括生长、肉质、皮毛、繁殖、抗病性能等，并筛选了与目标性状相关联的功能性基因和分子标记；繁殖技术研究少而散，包括人工授精、配种季节和光控管理等环境效应对家兔健康和生产力影响的研究。国外传统育种包括性状遗传评估、遗传与环境互作、评估方法等，在改进生长性能和肉质方面得到了提升；分子育种主要涉及生长、肉质、抗病及繁殖等性状，也进行了分子标记筛选与研究，但在皮毛性状方面研究相对欠缺；繁殖技术主要包括环境和添加物对家兔繁殖、精液冷冻等方面的影响研究，也取得了良好进展。本综述可为后期家兔育种研究和生产提供参考。     

奶牛CRP基因多态性与奶牛乳腺炎及乳质乳量的相关性研究及生物信息学分析

选用400头中国荷斯坦奶牛取血样400份,每份各2管。根据奶牛CRP基因序列设计2对引物,利用PCR-SSCP技术检测CRP基因的多态性,并用透射免疫比浊法测定奶牛血清CRP水平,然后运用SPSS等统计学软件对其与乳房炎及乳质、乳量的关系进行统计分析。结果显示,在第1对引物的扩增片段中检测到多态位点,出现3种基因型(AA、AB、BB),第2对引物的扩增片段中并未检测到多态位点。将第1对引物所扩片段的不同基因型与奶牛乳腺炎的相关性作关联分析,结果3种不同基因型奶牛的体细胞数、乳质率、蛋白率、305d产奶量均无显著差异,但患乳房炎奶牛个体全为AA型。而CRP含量检测结果可知患乳房炎的奶牛与正常个体之间的血清CRP含量并无显著差异。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

Enriched Topographic Microsites for Improved Native Grass and Forb Establishment in Reclamation

Low seed germination and seedling establishment are the greatest challenges for revegetation success. Topographic microsites are known to enhance seed germination and seedling establishment due to their unique soil properties and provision of shelter from elements and herbivores; soil amendments can supply organic matter and nutrients for plant establishment and growth when limited. We investigated the effect of three topographic microsites and six soil amendments and their additive effects on three disturbed grasslands in central and southern Alberta, Canada. Treatments were topographic microsites of mounds, pits, and flats, with and without amendments (erosion control blanket, hay, straw, manure, hydrogel, control) and were seeded with four native grasses and three native forb species. Seedling emergence and survival and soil temperature and water content were assessed over two seasons and plant cover over three seasons. The effect of microsites and amendments was not additive. The addition of erosion control blanket, hay, and straw to flat sites was just as productive as on topographic microsites. These amendments increased grass and forb emergence and buffered soil temperature. Mounds increased first year forb emergence and reduced over winter survival rates for grasses and forbs. Pits were not beneficial for revegetation. The effect of topographic microsites and amendments was influenced by site conditions.     

Research Progress on Diagnosis and Treatment of Canine and Feline Dermatophytosis

Canine and feline dermatomycosis is the common skin disease in small animal,which not only affects the appearance of the canine and feline, but also leads to itching or pain, and even increases the risk of dog and cat owners suffering from dermatomycosis. Dermatomycosis is harmful to health of animals and human beings. The dermatomycosis is difficult to identify, has long treatment cycle and high recurrence, and affected by regional or/and environmental factors, lead to its prevalent and brought great difficulties to the clinical diagnosis and treatment. Canine and feline dermatomycosis could be diagnosed according to illness history survey, clinical symptoms, isolation and identification of pathogens and histopathological examination. In order to curing canine and feline dermatomycosis effectively, systemic therapy combined with topical administration, scientific and rational use of antibiotics, and improvement of animal feeding management should be carried.     

Micronutrient Content of Certain Tropical Conventional and Unconventional Feed Resources of Southern India

The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies.     

Nutritional value and physiological effects of soya-free diets fed to rats during growth and reproduction

The objective of the study was to assess the effects of substitution milk and egg for soya products in breeding diets for rats, with concomitant decrease of the dietary protein level and supplementation with amino acids. Soya-containing (S) and two soya-free (NS and NSA) diets were evaluated as protein and energy sources, and their effects on reproductive performance during two cycles, and on the quality of the offspring were assessed. Organ weights were registered in females and blood parameters were determined in males. In the offspring males from S and NS groups, plasma LH, testosterone and prolactin levels were measured on the 22nd and the 60th day of life. The S diet contained more protein of smaller concentration of methionine and cystine and lower biological value than both NS and NSA diets and promoted similar post-weaning growth rate, similar body weight changes of dams during gestation and lactation and slightly lower mating efficiency. Within each reproductive cycle, the number and individual and total body weight of newborn and weanling pups did not differ but in two cycles mean number of neonates per litter and mean litter weight were significantly lower on S than on NSA diet. Plasma concentration of hormones did not differ in 22-day-old offspring males while in the older ones LH and prolactin levels were higher in animals fed on S than on NS diet. It is concluded that replacing soya protein by milk and egg protein with concomitant lowering dietary protein level and amino acid supplementation does not impair the growth rate and tends to improve reproductive performance. Feeding soya-free vs. soya-containing diets differentiates hormonal status of young males.     

反刍家畜瘤胃及肠道微生物研究进展

反刍家畜瘤胃及肠道微生物处于一个复杂的微生态系统,其多样性及营养代谢与该系统中的各项因素存在紧密联系。了解反刍家畜瘤胃及肠道微生物多样性变化和营养代谢机制,是通过人为干预手段改善动物福利、提高动物生产效率、提升动物产品质量、减少温室气体排放的基础。关于反刍动物瘤胃及肠道微生物多样性及营养代谢与单个环境因子的关系得到了广泛的研究并取得了一些进展,对指导生产实践和保护环境起到了一定作用。综述了近年来国内外反刍家畜瘤胃及肠道微生物多样性,以及微生物与宿主动物、日粮结构、环境因子相互作用及机制方面的研究进展,以期为探索瘤胃及肠道微生物在反刍动物营养代谢中的作用及其机制提供参考。