cDNA of Canine Heat Shock Protein 70 (HSP70)

Full-length canine HSP70 cDNA was sequenced and the expression of HSP70 mRNA was investigated. The full-length cDNA sequence of the HSP70 gene (2322 bp) contained a single long open reading frame (1920 bp) coding a protein of 640 amino acids. The amino acid sequence of the canine HSP70 gene shared about 90-95% sequence similarity with bovine, human and mouse HSP70 proteins. Southern blot analysis with HSP70 probe gave three distinct bands of 9.4 kb, 5 kb and 4.4 kb in BamHI digests and two distinct bands of 19 kb and 4 kb in EcoRI digests. Canine HSP70 mRNA was detectable in canine peripheral blood mononuclear cells and stomach but not in liver, kidney, spleen, small intestine, large intestine and skin of dogs.     

Effect of bilateral cecal ligation on use of dietary energy in growing chickens

In order to determine the role of the cecum on energy use in growing chickens, metabolizability of the dietary energy and energy expenditure were examined in the week following bilateral ligation and washing out of the cecum. Apparent metabolizable energy (AME) values were 14.30 and 13.69 kJ/g air‐dry matter for sham‐operated and cecally ligated chickens, respectively. These values were found to be significantly different (P < 0.05). Although AME intake and fasting heat production were decreased by cecal ligation, the distribution of AME (measured as fasting and feeding heat production, as well as heat increment and energy retention, as a proportion of AME intake) was not affected. These results suggest that the cecum helps chickens extract AME from corn‐soybean type diets with little, if any, effect on AME use, based on the present study of growing chickens in the week following cecal ligation.     

Assessment of Uterine Vascular Perfusion During the Estrous Cycle of Mares in Connection to Circulating Leptin and Nitric Oxide Concentrations

The objective of this study was to find difference in vascular perfusion of uterine horns or uterine body throughout the estrous cycle and their relation to circulating nitric oxide and leptin concentrations. Five cyclic mares were subjected to transrectal Doppler ultrasonography and blood sampling for 18 days. Area of color and power Doppler modes was measured in pixels. Day (P = .0001) of the estrous cycle and ovulation (P = .0001) influenced uterine blood flow. Uterine body blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day 0 (day of ovulation), and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until days 12 and 10, respectively. Conversely to the contralateral uterine horn, ipsilateral uterine horn blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day −1, and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until day 10. Nitric oxide concentrations (P = .0001) attained two major peaks; the first on day −3 and the other persisted from day 2 until day 5. Leptin concentrations increased (P > .001) with a maximum value on day 0 and then decreased until a minimum value on day 9. In conclusion, during the estrous cycle, ipsilateral uterine horn and uterine body blood vessels had similar blood flow. Both leptin and nitric oxide played a role during follicle growth, ovulation, and corpus luteum development and modulated uterine blood flow before and after ovulation.     

鸡Hsc70基因5′侧翼区SNP与耐热性状的相关分析

为研究Hsc70基因作为鸡抗病育种中一种分子标记的可能性,本试验利用克隆测序技术对清远麻鸡、广东温氏矮脚黄鸡、灵山鸡和隐性白羽洛克鸡4个品种共20个个体Hsc70基因的5′侧翼区进行多态性筛查,发现10处变异,包括8个SNPs和2个插入/缺失突变。通过分析,选取C.-521A〉C位点,利用PCR-RFLP方法分析了此位点在灵山鸡和隐性白羽洛克鸡群体中的基因型分布,所得分型结果与测定的耐热性状（T3、皮质酮、CD3＋、CD4＋和CD8＋）进行关联分析,结果表明：常温状态下（15℃）,在隐性白羽洛克鸡群体中,位点C.-521A〉C与CD3＋、CD4＋和CD8＋显著相关（P〈0.05）,其中AA基因型个体CD3＋值显著低于AC基因型个体,而CD4＋与CD8＋值显著高于AC基因型个体;热应激状态下（35℃）,在灵山鸡群体中,位点C.-521A〉C与CD8＋极显著相关（P〈0.01）,CC基因型个体CD8＋值极显著高于AA和AC基因型。     

产蛋下降综合征（EDS－76）病毒DNA探针的制备及应用研究

用纯化的产蛋下降综合征（ＥＤＳ－７６）病毒（Ｈ９１毒株）悬液提取的核酸，经琼脂糖凝胶电泳只出现１条分子量较大、背景清晰的核酸带。用ＢａｍＨＩ酶消化产生４个片段，大小分别为１７ｋｂ、１０ｋｂ、４．０ｋｂ和２．０ｋｂ。将其中的２．０ｋｂ片段克隆进ｐＵＣ１８ＤＮＡ载体中，重组质粒ＤＮＡ用ｄｉｇｏｘｉｇｅｎｉｎ标记作为ＤＮＡ探针，在ｄｏｔｂｌｏｔ中该探针不与鹅胚尿囊液核酸抽提物、马立克氏病病毒（ＭＤＶ）ＤＮＡ、鸡传染性贫血病毒（ＣＡＶ）ＤＮＡ、ＳＰＦ鸡基因组ＤＮＡ等核酸反应，只与３株ＥＤＳ病毒（ＥＤＳ标准毒株ＡＶ－１２７、ＥＤＳＨ９１毒株和从健康鹅体中分离的ＥＤＳＹ８１毒株）ＤＮＡ呈阳性反应。人工感染产蛋母鸡和雏鸡后２４ｈ即可用该探针从泄殖腔棉拭子样品中检测出ＥＤＳ病毒ＤＮＡ，在感染后３５ｈ仍能从部分感染鸡样品中检出。对部分探针检测阳性鸡的泄殖腔棉拭子样品用ＳＰＦ鸡胚分离病毒，并用ＨＡ和ＨＩ试验检测分离病毒的特异性；在感染过程中，同时检测了血清中ＨＩ抗体的消长情况。结果表明，人工感染鸡排毒至少可以维持２个月左右，而且血清中ＨＩ抗体即使很高，也不能阻止感染鸡的排毒。     

Effect of oxytocin infusion on luteal blood flow and progesterone secretion in dairy cattle

The objective of this study was to investigate the effects of oxytocin infusion on corpus luteum (CL) function during early to mid-diestrus by measuring luteal size (LS) and luteal blood flow (LBF) along with plasma levels of progesterone (P4) and prostaglandin metabolites (13,14-dihydro-15-keto-prostaglandin F_2α, PGFM). On day (D) 7 of the estrus cycle (D1 = ovulation), seven cows received 100 IU of oxytocin (OXY) or placebo (PL) following a Latin square design. LS and LBF increased in both groups over time and no differences were observed between the groups. PGFM did not differ either within the groups over time or between the groups at any time point. P4 of the OXY group was higher compared to that of the the PL group 360 min after the infusion (p = 0.01) and tended to be higher at the time points 450 min, 48 h, and 72 h (all p = 0.08). Results from this study support the hypothesis that OXY is not directly involved in the mechanism(s) governing blood flow of the CL and has no remarkable effects either on luteal size or P4 and PGFM plasma levels. Further investigation is needed to elucidate the role of OXY in CL blood flow during early and late luteal phases.     

持续高温应激对肥育猪生产性能、胴体性状、背最长肌营养物质含量及肌纤维特性的影响

本试验旨在研究持续30℃的环境高温应激对肥育猪生产性能、胴体性状、背最长肌质构分析、营养物质含量及肌纤维特性的影响.试验选取8窝共24头体重为(79.0±1.5) kg的“杜长大”三元阉公猪,其中每3头来自同一窝的猪只随机分到3组,每组8个重复,每个重复1头,3组分别为对照组(22℃适温+自由采食)、高温应激组(30℃高温+自由采食)和采食量配对组(22℃适温+采食量配对),试验期21d.结果表明:1)持续高温应激显著降低肥育猪平均日增重(ADG) (P＜0.05),极显著降低肥育猪平均日采食量(ADFI) (P＜0.01).2)3组之间的肥育猪胴体重、胴体长、平均背膘厚和皮厚均差异不显著(P＞0.05),高温应激组肥育猪背最长肌大理石花纹评分在宰后24 h有低于对照组的趋势(P＜0.10).3)高温应激组肥育猪背最长肌的剪切力极显著高于对照组和采食量配对组(P＜0.01).与采食量配对组相比,高温应激组肥育猪背最长肌胶黏性和硬度有降低的趋势(P＜0.10).4)高温应激组和采食量配对组肥育猪背最长肌粗蛋白质含量显著高于对照组(P＜0.05),且2组的背最长肌肌内脂肪(IMF)含量极显著低于对照组(P＜0.01).5)高温应激组肥育猪背最长肌Ⅰ型肌纤维含量有低于采食量配对组的趋势(P＜0.10),采食量配对组的Ⅱx型肌纤维含量显著低于对照组(P＜0.05).由此可见,肥育后期高温应激会改变猪表观肉品质和肌肉营养物质含量.     

小反刍兽疫病毒及山羊痘病毒N-ITR基因二联实时荧光定量RT-PCR检测方法的建立

Peste des petits ruminants virus (PPRV) and goat pox virus (GTPV) are the causative agents of two kinds of goats’ diseases-peste des petits ruminants and goat pox which can cause disaster economic losses. In order to detect the two viruses simultaneously and quickly, two sets of primers and relative probes were designed based on the nucleoprotein (N) gene of PPRV and the inverted terminal repeat (ITR) segment of GTPV, respectively. In order to work together in the same reaction, the probes were labeled with different fluorescent materials 5′FAM-TAMRA3′and 5′JOE-Eclipse3′,respectively. Results showed that the duplex Real-time RT-PCR assay was identified to be specific for PPRV and GTPV only and specific fluorescent signal could be detected, but the related viruses including fowl pox virus（FPV）and canine distemper virus (CDV) had no specific fluorescent signal. Positive recombinant plasmids (PPRV pMD18-T-N and GTPV pMD18-T-ITR) were built and used for positive quantitative templates to establish duplex standard curves. The developed assay based on the probe N-ITR was found to be highly specific and sensitive with a detection limit of 10² copies/μL cDNA and 10³ copies/μL DNA for PPRV and GTPV, respectively. Finally, the duplex Real-time RT-PCR assay for simultaneous detection of PPRV and GTPV was established preliminarily in the study.