牛胚胎移植技术的产业化研究

本文探讨了引进国外优质奶、肉牛冷冻胚胎建立核心群，以及供体牛选择、超数排卵、胚胎冷冻保存和分割等相关技术，为牛胚胎移植产业化奠定基础。     

Changes in fox squirrel anti-predator behaviors across the urban–rural gradient

Predator stimuli created by humans in the urban environment may alter animals’ anti-predator behaviors. I hypothesized that habituation would cause anti-predator behaviors to decrease in urban settings in response to humans. Additionally, I hypothesized that populations habituated to humans would show reduced responses to other predator stimuli. I observed three populations of squirrels (urban, suburban and rural) responses to human approaches, red-tailed hawk vocalizations (Buteo jamaicensis) and coyote (Canis latrans) vocalizations. Mahalanobis distances of anti-predator behaviors in response to human approaches were consistent with the urban–rural gradient. Flight initiation distances (X ² = 26.33, df = 2, P < 0.001) and amount of time dedicated to anti-predator behavior (X ² = 10.94, df = 2, P = 0.004) in response to human approaches were also consistent with the urban–rural gradient. Supporting the habituation hypothesis, naive juvenile squirrels increased flight initiation distances (X ² = 35.89, df = 1, P < 0.001) and time dedicated to anti-predator behaviors (X ² = 9.46, df = 1, P = 0.002) relative to adult squirrels in the same urban environment. Time dedicated to anti-predator behaviors differed among all three sites in response to both coyote (X ² = 9.83, df = 2, P = 0.007) and hawk (X ² = 6.50, df = 2, P = 0.035) vocalizations. Responses to both vocalizations on rural sites (coyote = 45%, hawk = 55%) greater than twice that found on the urban sites (coyote = 11%, hawk = 20%). This is possibly the first case of a transfer of habituation demonstrated under field conditions.     

湖羊胚胎和精液超低温保存效果的评价

【目的】 检验长期保存在国家家畜基因库的湖羊冷冻胚胎和冷冻精液的质量,评价超低温冷冻保存技术保种的效果。【方法】 对保存于国家家畜基因库的冷冻胚胎（保存20年）和冷冻精液（保存20和30年）进行复苏,进行相应鉴定后分别进行胚胎移植和人工授精,同时以0年冷冻精液和新鲜精液为对照,测定其受胎率和后代的羔皮性能、生长性能及繁殖性能,检验保存效果。【结果】 本试验共解冻胚胎36枚,其中,A级胚胎22枚,B级胚胎7枚,C级胚胎5枚,D级胚胎2枚,冷冻胚胎复苏利用率达到94.44%（34/36）,A级胚胎率达到61.11%（22/36）,移植34枚,产羔17只,冷冻胚胎复苏移植产羔率达50.00%;保存30年的冷冻精液复苏后平均精子活力达35%,受胎率为58.57%,平均产羔1.90只;保存20年冷冻精液复苏后平均精子活力达31%,受胎率为53.66%,平均产羔1.95只;胚胎复苏移植羊、30和20年冷冻精液后代体型外貌鉴定均符合纯种湖羊特征,没有畸形个体;羔羊一级羔皮率分别是31.25%、42.03%和23.81%,保持了当年湖羊的羔皮特性;生长发育及繁殖性能与现有湖羊群体性能基本保持一致。【结论】 利用超低温冷冻技术长期保存湖羊冷冻胚胎和冷冻精液的方法是可行的,对地方品种保种具有重要意义。     

Low levels of haptoglobin and putative amino acid sequence in Taiwanese Lanyu miniature pigs

Porcine haptoglobin (Hp) is an acute phase protein. Its plasma level increases significantly during inflammation and infection. One of the main functions of Hp is to bind free hemoglobin (Hb) and inhibit its oxidative activity. In the present report, we studied the Hp phenotype of Taiwanese Lanyu miniature pigs (TLY minipigs; n=43) and found their Hp structure to be a homodimer (beta-alpha-alpha-beta) similar to human Hp 1-1. Interestingly, Western blot and high performance liquid chromatographic (HPLC) analysis showed that 25% of the TLY minipigs possessed low or no plasma Hp level (<0.05 mg/ml). The Hp cDNA of these TLY minipigs was then cloned, and the translated amino acid sequence was analyzed. No sequences were found to be deficient; they showed a 99.7% identity with domestic pigs (NP_999165). The mean overall Hp level of the TLY minipigs (0.21 +/- 0.25 mg/ml; n=43) determined by enzyme-linked immunosorbent assay (ELISA) was markedly lower than that of domestic pigs (0.78 +/- 0.45 mg/ml; p<0.001), while 25% of the TLY minipigs had an Hp level that was extremely low (<0.05 mg/ml). In addition, the initial recovery rate (first 40 min) in the circulation of infused fluorescein isothiocyanate (FITC)-Hb was significantly higher in the TLY minipigs with extremely low Hp levels than those with high levels. This data suggests that the low concentration of Hp-Hb complex is responsible for the higher recovery rate of Hb in the circulation. TLY minipigs have been used as an experimental model for cardiovascular diseases; whether they can be used as a model for inflammatory diseases, with Hp as a marker, remains a topic of interest. However, since the Hp level varies significantly among individual TLY minipigs, it is necessary to prescreen the Hp levels of the animals to minimize variation in the experimental baseline. The present study may provide a reference value for future use of the TLY minipig as an animal model for inflammation-associated diseases.     

绵羊FGF10基因克隆、序列分析及其在毛囊发育中的表达分析

本试验旨在获得中国美利奴羊成纤维细胞生长因子10（fibroblast growth factor 10,FGF10）基因的编码区（CDS）全长序列并进行生物信息学分析,随后对FGF10基因在中国美利奴羊毛囊发育过程中的表达特征进行分析,明确其在中国美利奴羊毛囊发育过程中的表达模式,为进一步研究FGF10 mRNA表达水平与中国美利奴羊毛囊生长发育的表达调控机制奠定理论基础。采用PCR扩增获得中国美利奴羊FGF10基因CDS,并克隆到zero PCR@^TM-Blunt进行测序验证;利用实时荧光定量PCR技术检测FGF10在中国美利奴羊毛囊发育过程中的表达差异。结果表明,绵羊FGF10基因CDS长度为696 bp（序列上传GenBank,获得登录号：MT872422）,编码231个氨基酸,与牛和山羊的氨基酸序列同源性达100%,存在1个信号肽和1个跨膜结构域,其为分泌通路信号蛋白;实时荧光定量PCR分析表明,FGF10基因在中国美利奴羊毛囊发育过程中均表达,在毛囊发育第85天表达最高,显著高于其他毛囊发育时期（P<0.05）。本研究获得中国美利奴羊FGF10基因完整的编码区序列和毛囊发育过程中的表达特征,生物信息学分析发现,FGF10基因编码区序列具有物种间的保守性,同时FGF10在绵羊毛囊不同发育阶段的皮肤组织中表达,由此表明,FGF10基因可能在绵羊毛囊的生长发育过程中发挥重要的生物学作用。     

马铃薯金线虫和白线虫DNA特异扩增产物的克隆与鉴定

通过采用Fullaondo（1999）研究的特异引物检测方法,对来自英国和比利时的两个马铃薯金线虫（Globodera rostochiensis）种群和英国的一个马铃薯白线虫（G.pallida）种群的DNA进行扩增,其产物以大肠杆菌（Eschetichia coli）为受体菌,经过连接、转化和克隆纯化,筛选获得阳性克隆.经PCR鉴定和测序,英国和比利时的马铃薯金线虫种群的特异性谱带分别是357bp和350bp,而英国的马铃薯白线虫的谱带是782bp,与Fullaondo等人研究报道结果基本相符,从而鉴别出来自不同地理种群的马铃薯金线虫和白线虫.     

Skin and coat traits in sheep in Brazil and their relation with heat tolerance

The objective of this study was to evaluate and compare physical parameters in groups of sheep with different phenotypic characteristics in the Centre-west region of Brazil. Five groups of sheep, with nine animals per group, were selected, three groups of Santa Inês animals with different coat colours (white, brown and black), one group with crossbred animals (Santa Inês × Bergamasca) and one group with animals of the Bergamasca breed. The following traits were evaluated: coat thickness, number and length of hair, pigmentation level in the coat and the skin as well as the percentage area of sweats glands in the skin tissue, carried out by histological analysis. The number of hairs and the area of sweats glands were not significantly different between the evaluated groups. The Bergamasca breed showed low pigmentation of the skin and long hairs. The levels of pigmentation of the hair and of the skin were highly correlated. Between the Santa Inês groups, the group with white hair showed the better parameters for heat adaptation, while the brown hair group showed the lower heat adaptation when compared with another hair breed groups.     

Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE₂ was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0?×?10⁵ cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.     

按不同氨基酸模式配制肉鸭日粮的比较研究

试验比较了不同氨基酸模式对肉鸭生产性能的影响。将320只1日龄的樱桃谷鸭随机分为4组,每组80只,设4个重复,每个重复20只,公母各半,分别饲喂四种日粮(按四种氨基酸模式配制),试验期21天。结果表明:日粮的氨基酸模式为Lys∶Met∶Trp∶Thr=100∶45∶21∶62时,其2周和3周体重高于其它各组,但差异不显著(P>0.05),且F/G和血清尿酸水平最低。     

基于深度学习的病虫害智能化识别系统

我国农作物种植覆盖面广、分散度高,病虫害发生种类多、区域性发生规律复杂,传统的人工鉴定技术从效率、能力与精度方面均难以满足新形势下重大病虫测报要求。针对这一实践需求,以测报灯下害虫图像数据库(约18万张)、田间病虫害图像数据库(约32万张)为基础,构建了基于深度学习方法的病虫害种类特征自动学习、特征融合、识别和位置回归计算框架,并研发了移动式病虫害智能化感知设备和自动识别系统。通过近2年的精确度和实操运行效率检验,该系统在自然状态下对16种灯下常见害虫的识别率为66%～90%,对38种田间常见病虫害(症状)的识别率为50%～90%。随基础数据库的不断丰富、神经网络深层特征提取的不断完善,该系统有望进一步提高识别准确率,从而真正实现田间病虫害识别自动化、智能化和高效率。