1例藏獒三氮脒中毒的生化检验及治疗

用三氮脒以3.5 mg/kg的剂量对患有附红细胞体病的藏獒进行肌肉注射,隔日用同等剂量的三氮脒肌肉注射,24 h后,藏獒发生以神经症状为主的中毒现象,并表现出食欲废绝,后肢发软,四肢抽搐呈划水状,卧地不起,昏迷,体温正常临床症状。经血清生化检验,藏獒的尿素氮、肌酸激酶等的生化指标明显升高,血糖降低占病例的100%。经过5 d的药物治疗后,中毒藏獒基本恢复健康。     

不同日龄滩羊皮肤组织化学特征比较

为比较2、35日龄滩羊皮肤毛囊的发育特点与血管内皮生长因子（vascular endothelial growth factor，VEGF）和血管内皮生长因子受体2（vascular endothelial growth factor receptor 2，VEGFR2）的分布特征，探究出生后滩羊被毛生长发育的变化特点，试验应用常规HE染色及改良Masson胶原纤维染色、Gomori银氨法染色、磷钨酸染色等特殊染色观察2与35日龄滩羊皮肤组织结构特点；应用免疫组织化学法结合免疫荧光染色法观察VEGF及VEGFR2在2与35日龄滩羊皮肤组织中的分布定位，并用IPP图像分析软件进行定量分析。结果显示：与2日龄滩羊皮肤组织比较，35日龄滩羊表皮与真皮间界限更加清晰，毛囊结构发育完整；毛囊密度显著降低（P<0.05）；胶原纤维与弹性纤维含量增加，形成网格状分布。免疫组化及免疫荧光结果显示，VEGF及VEGFR2在滩羊皮肤表皮及毛囊外根鞘、皮脂腺上均有表达。统计表明，VEGF及VEGFR2在2日龄滩羊皮肤组织中的表达量均显著高于35日龄（P<0.05）。综合上述结果，滩羊毛囊发育过程中，胶原纤维和弹性纤维增加明显；VEGF与VEGFR2通路在毛囊角质形成中起直接调节作用。     

Effects of Grazing Intensity on Soil and Vegetation Properties in a Mediterranean Rangeland

Grazing modifies the aboveground soil and plants of rangeland ecosystems. This study was conducted to determine the effects of various grazing intensity levels on soil bulk density, soil pH, soil electrical conductivity (EC), soil total organic carbon (TOC), aboveground biomass production, plant community richness (N), and vegetation diversity (H) in Gian Rangeland, Hamedan Province, Iran. Soil and vegetation samplings were conducted based on the randomized systematic method. Soil and vegetation samplings were carried out along 100-m-long transects placed at 100, 200, 400, 800, and 1600 m running perpendicular to six sheep corrals. The effects of various grazing intensities on studied parameters were detected by analysis of variance (ANOVA) (at α = 0.05). Results of this study indicated the soil bulk density, EC, TOC, aboveground biomass production, plant community richness, and vegetation diversity values were altered significantly at different distances from the sheep corrals. However, soil pH was not significantly affected by various grazing intensities.     

绵羊Musclin基因真核表达载体的构建及其对肌细胞糖代谢和胰岛素作用的影响

为了研究绵羊Musclin对糖代谢和胰岛素作用的影响,本试验运用生物信息学软件对其结构特征进行预测,并构建绵羊Musclin基因真核表达载体。分4组对绵羊胎儿成肌细胞进行处理：空载体组（pcDNA3.1）、Musclin过表达组（pcDNA3.1-Musclin）、空载体+胰岛素组（pcDNA3.1+Insulin）、Musclin过表达+胰岛素组（pcDNA3.1-Musclin+Insulin）。然后分别在分化48和72 h时收集细胞,检测Musclin对正常或添加胰岛素培养的分化状态绵羊肌细胞糖代谢的影响,并利用荧光定量PCR检测相关基因表达的变化。酶切鉴定和测序结果表明,成功构建了表达载体pcDNA3.1-Musclin。生物信息学分析表明,绵羊Musclin蛋白含有一个信号肽,并在28氨基酸位点处存在一个酶切位点。亚细胞定位在胞外,属于分泌蛋白。其二级和三级结构以α螺旋和无规则卷曲为主。细胞试验表明,绵羊Musclin基因过表达明显减少了正常或胰岛素处理的绵羊肌细胞糖原含量并增加了培养液中葡萄糖含量。诱导分化72 h时,过表达Musclin抑制了Musclin过表达组和Musclin过表达+胰岛素组绵羊肌细胞中IRS1、AKT1、AKT2、GLUT1、GLUT4基因mRNA表达,并促进了GSK3β mRNA表达;而诱导分化48 h时,只有GLUT4基因的表达受到抑制。综上所述,绵羊Musclin基因过表达可以影响肌细胞糖代谢并减弱胰岛素的作用,二者相互协调以维持糖代谢稳态。其作用机制涉及到上述相关基因的表达变化,且与绵羊成肌细胞的分化状态有关。     

戴瑞奶绵羊产奶性状的全基因组关联分析

旨在通过对奶绵羊产奶性状的全基因组关联分析（GWAS）,寻找和定位与奶绵羊产奶性状相关的遗传标记和功能基因。本研究以135只戴瑞奶绵羊为试验材料,基于全基因组重测序技术,通过SAMTOOLS进行单核苷酸多态性（SNP）检测,使用PLINK v1.90进行质控,利用GEMMA v 0.98.1的混合线性模型对质控结果进行奶绵羊产奶性状相关的全基因组关联分析。结果表明,有1个SNP与泌乳后90天日均产奶量在全基因组范围内显著相关,8个SNPs达到潜在显著关联,相关的候选基因包括TRNAQ-CUG-2、LOC114117240、ACADL、MYL1、CHD6、SLCO3A1;有2个SNPs与150天日均产奶量达潜在显著关联,相关的候选基因包括PRMT6、RNF180;有2个SNPs与泌乳周期达潜在显著关联,相关的候选基因包括PRMT6、TRNAW-CCA-68、TRNAS-GGA-61。进一步基因功能分析推测,ACADL、SLCO3A1可能是影响奶绵羊产奶性状的候选基因。本研究为奶绵羊产奶性状的分子机制解析提供了一定的基础,为我国奶绵羊新品种培育提供了一定的理论参考。     

Optimizing injection time of GFP plasmid into sheep zygote

Microinjection of exogenous DNA into the cytoplasm of matured oocytes or zygotes is a promising technique to generate transgenic animals. However, the data about the microinjection time and procedure in sheep are limited and have not treated in detail. To obtain more in-depth information, the Sarda sheep oocytes from abattoir-derived ovaries were subjected to IVM and IVF. Then, the GFP plasmid as a reporter gene was injected into the cytoplasm of MII oocytes (n: 95) and zygotes at different post-insemination intervals (6–8 hpi, n: 120; 8–10 hpi, n: 122; 10–12 hpi, n: 110 and 12–14 hpi, n: 96). There were no significant differences in the cleavage rates between the groups. However, blastocyst rate of injected zygotes at all-time intervals was significantly lower than injected MII oocytes and control group (p < 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, p  < 0.01). Among these, the proportion of mosaic embryos was high and two of those embryos developed to the blastocyst stage. In conclusion, we settled on the cytoplasmic microinjection of GFP plasmid at 8–10 hpi as an optimized time point for the production of transgenic sheep and subsequent experiments.     

复合双歧杆菌制剂对肉羊生长性能、养分表观消化率及血液生化指标的影响

为研究复合双歧杆菌制剂对肉羊生长性能、养分表观消化率及血液生化指标的影响,试验选择健康、体重相近的杜寒杂交肉羊60只随机分成4组,每组15个重复,每个重复1只,1组为对照组饲喂基础日粮,试验2、3、4组分别在基础日粮中添加2.5%、5.0%、10%的复合双歧杆菌制剂,预试验7 d,试验期56 d。结果表明:(1)试验3、4组的平均日增重较1组比分别提高13.5%、9.3%(P<0.05),料重比较1组比分别降低10.3%、9.4%(P<0.05),试验2、3、4组的干物质采食量均高于1组(P>0.05);(2)试验2、3、4组的干物质、钙、磷、酸性洗涤纤维表观消化率均高于1组(P>0.05),试验3、4组的粗蛋白质、中性洗涤纤维消化率较1组比分别提高17.2%、15.3%、12.5%、9.1%(P<0.05);(3)试验3、4组血清中的总蛋白(TP)含量较1组比分别提高16.0%、12.9%(P<0.05),试验2、3、4组血清中的白蛋白(ALB)含量均高于1组(P>0.05),试验2、3、4组血清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)含量均低于1组(P>0.05),试验3、4组血清中尿素氮(BUN)含量较1组比分别降低15.4%、13.8%(P<0.05)。综上,不同水平的复合双歧杆菌制剂可以提高肉羊生长性能、养分表观消化率,且对血液生化指标无明显影响,以5.0%添加为适宜。     

计算机视觉技术在智慧羊场中的研究进展

智慧羊场具有科学化、标准化管理的优点,是羊场转型发展的趋势。计算机视觉技术是人工智能的核心,可以实现非接触性、自动化、实时监测羊的个体信息。本文总结并分析了计算机视觉在羊场的个体识别方法、行为识别方法、体尺与体重估测、疾病监测的研究现状,并指出计算机视觉在智慧羊场的进一步发展趋势。 [关键词] 计算机视觉|智慧羊场|个体识别|行为识别|体尺与体重估测|疾病监测     

妊娠后期营养限制对蒙古绵羊胎儿生物原胺类神经递质的影响

本试验旨在研究妊娠后期营养限制对蒙古绵羊胎儿生物原胺类神经递质的影响。选择健康、体况相近的蒙古绵羊18只,对其进行同期发情、配种后,从妊娠第90天开始,随机分为3个组:营养限制1组[NG1组,n=6,代谢能(ME)=0.175 MJ/(kg BW 0.75·d)]、营养限制2组[NG2组,n=6,ME=0.330 MJ/(kg BW 0.75·d)]和对照组[CG组,n=6,ME=0.670 MJ/(kg BW 0.75·d)],在妊娠第140天时屠宰,取各组胎儿及其脑组织和血液。结果显示:NG1组胎儿的脑重较CG组显著增高(P<0.05),并且NG1组胎儿脑中5-羟色胺(5-HT)(P<0.01)、肾上腺素(EPI)(P<0.05)含量显著或极显著高于CG组,去甲肾上腺素(NA)的含量极显著低于CG组(P<0.01)。NG2组胎儿脑中5-HT含量极显著高于CG组(P<0.01),而NA(P<0.01)、多巴胺(DA)(P<0.05)含量显著或极显著低于CG组。另外,NG1组胎儿血浆中5-HT(P<0.01)、EPI(P<0.01)、NA(P<0.05)、DA(P<0.01)含量显著或极显著低于CG组;NG2组胎儿血浆中5-HT(P<0.01)、NA(P<0.01)、DA(P<0.05)含量显著或极显著低于CG组。由此得出,妊娠后期营养限制使得蒙古绵羊胎儿脑中5-HT、EPI含量升高和NA含量降低,血浆中5-HT、EPI、NA、DA含量降低。     

Toll-like receptor signaling is changed in ovine lymph node during early pregnancy

Toll-like receptors (TLRs) participate in regulation of adaptive immune responses, and lymph nodes play key roles in the initiation of immune responses. There is a tolerance to the allogenic fetus during pregnancy, but it is unclear that expression of TLR signaling is in ovine lymph node during early pregnancy. In this study, lymph nodes were sampled from day 16 of nonpregnant ewes and days 13, 16, and 25 of pregnant ewes, and the expressions of TLR family (TLR2, TLR3, TLR4, TLR5 and TLR9), adaptor proteins, including myeloid differentiation primary-response protein 88 (MyD88), tumor necrosis factor receptor associated factor 6 (TRAF6), and interleukin-1-receptor-associated kinase 1 (IRAK1), were analyzed through real-time quantitative polymerase chain reaction, Western blot, and immunohistochemistry analysis. The results showed that mRNA and protein levels of TLR2, TLR3, TLR4, TRAF6, and MyD88 were upregulated in the maternal lymph node, but TLR5, TLR9, and IRAK1 were downregulated during early pregnancy. In addition, MyD88 protein was located in the subcapsular sinus and lymph sinuses. Therefore, it is suggested that early pregnancy induces changes in TLR signaling in maternal lymph node, which may be involved in regulation of maternal immune responses in sheep.