血液能量平衡指标对高产奶牛产后卵巢静止的影响

本试验旨在确立能量平衡对奶牛产后卵巢静止的作用,为今后预防该病提供科学依据。从黑龙江省某集约化牛场随机选取产后14~21d奶牛80头(年龄2~4岁,胎次1~3次,泌乳量(34.82±9.51)kg/d),跟踪观测至产后60~90d,进行奶牛发情鉴定,结合B超观察与生殖激素检测从观测奶牛中选出卵巢静止与发情奶牛各16头。对2组奶牛产后14~21d和60~90d的血浆能量代谢及肝功能指标进行了统计比较。结果显示:产后60~90d卵巢静止奶牛血浆β-羟丁酸和游离脂肪酸含量高于发情奶牛(P0.05),血浆促卵泡素、雌二醇和孕酮浓度极显著低于发情奶牛(P0.01);根据受试者工作特征曲线分析确定产后14~21d血浆β-羟丁酸0.69 mmol/L、游离脂肪酸0.52mmol/L作为产后卵巢静止预警值,灵敏度和特异度分别为85.7%和83.30%,92.3%和86.70%,产后60~90d2项指标的预警值分别为0.42,0.17mmol/L。奶牛产后14~21d与60~90d血浆β-羟丁酸和游离脂肪酸浓度超过预警值,奶牛产后发生卵巢静止的风险会增大。     

卵巢静止奶牛血液矿物质及生化指标变化研究

为了给牧场防控奶牛卵巢静止疾病提供科学依据,试验选取产后45～60天卵巢静止的和正常发情的中国荷斯坦奶牛各15头,采用抽血检测的方法分析了卵巢静止奶牛血液中矿物质及生化指标的变化情况。结果表明:与正常发情组奶牛相比,卵巢静止组奶牛日泌乳量显著提高(P0.05),发情次数显著减少(P0.05),卵泡直径极显著减小(P0.01);丙氨酸氨基转移酶(ALT)活性显著提高(P0.05),天门冬氨酸氨基转移酶(AST)活性极显著提高(P0.01);血中矿物质镁(Mg)含量显著降低(P0.05);另外,卵巢静止组奶牛血中雌二醇(E_2)浓度极显著降低(P0.01)。说明产后卵巢静止奶牛血液能量、肝功指标、矿物元素及生殖激素等发生了明显变化。     

高产奶牛产后卵巢静止血液临床病理学变化及其预警评估

选择黑龙江省2个牛场高产奶牛,在产后60~90d调查卵巢静止,检测发情和卵巢静止奶牛生殖激素,能量代谢,肝功能和矿物质等指标,通过Pearson相关分析、二元Logistic分析和ROC分析对牛场产后卵巢静止进行预警评估。结果显示:2个牛场(Ⅰ,Ⅱ)卵巢静止发病率分别为29.5%,47.2%,主要由于产后早期能量负平衡。2个牛场卵巢静止组与发情组相比,血浆中β-羟丁酸(BHBA)、游离脂肪酸(NEFA)和门冬氨酸氨基转移酶(AST)均显著的升高。牛场Ⅰ,当产后60~90d奶牛血浆中BHBA0.94 mmol/L、NEFA0.56 mmol/L、AST95.5U/L时;牛场Ⅱ,当奶牛产后60~90d血浆中BHBA1.00mmol/L、NEFA0.585mmol/L、AST103.5U/L时;奶牛患卵巢静止的风险增高。结果表明,奶牛产后发生的能量负平衡可扰乱奶牛发情周期的生殖激素分泌,是引起2个牛场奶牛产后卵巢静止的主要病因。奶牛泌乳早期血浆中NEFA、BHBA和AST等指标可用于产后卵巢静止的风险预警。     

基于iTRAQ技术的卵巢静止奶牛血清差异蛋白分析

旨在利用iTRAQ技术探究卵巢静止奶牛血清差异蛋白。本试验在奶牛产后60～90 d,根据发情表现、直肠检查和B超检查,按判定标准将试验奶牛分为卵巢静止组(IO)和正常发情对照组(CON),每组50头,采集血清。应用同位素标记绝对定量(iTRAQ)/质谱(MS)联用技术筛选差异蛋白,并应用免疫印迹以及ELISA方法鉴定奶牛卵巢静止血清差异蛋白。结果表明,试验共获得61种血清差异表达蛋白。GO分析表明,生物学过程包括34个注释,分子功能包括15个注释,细胞组成包括13个注释。蛋白质网络互作分析共得到57个节点,68条蛋白互作注释,蛋白质互作富集P<10^-16,得到11个信号代谢通路。经过筛选发现,糖酵解/糖异生、氨基酸的生物合成、胰高血糖素信号通路,维生素的消化与吸收可能与卵巢静止的发生存在相关性。与卵巢静止相关差异表达蛋白共有14种:其中GPX3、SCGB1D和PKM2表达下调,ADIPOQ、AHSG、APOA4、FETUB、ALDOB、SPAM1、LDHB、RBP4、IGFBP2、ITIH3及GLYCAM1表达上调。ADIPOQ、IGFBP2和RBP4通过生殖激素生物过程影响卵泡发育,GPX3通过氧化应激而影响卵泡发育。     

奶牛产后肝功指数与卵巢静止的关系及其预警作用

为明确奶牛产后卵巢静止与肝脏功能的内在联系,以肝功指数(liver function index, LFI)为自变量,分为低LFI组(n=30头)和高LFI组(n=30头),采用B超、队列研究的M-Hχ²检验分析、双变量相关分析、二元Logistic回归模型和ROC分析的方法,分析60头奶牛产后21 d血浆中氧化应激、能量代谢、矿物质、激素等指标和50～55 d卵泡发育状况,并确定LFI对奶牛卵巢静止的风险预测值。结果显示,低LFI的奶牛在产后21 d的SOD、GSH、GH、Glu极显著降低(P<0.01);抑制羟自由基能力、E₂、P₄、IGF-1显著降低(P<0.05);BHBA极显著升高(P<0.01)、NEFA显著升高(P<0.05)。并且,低LFI使奶牛产后卵巢静止发生的风险增高2.67倍,LFI对奶牛卵巢静止的最佳预警值为LFI<-3.15,有较高的诊断意义。     

苜蓿素对脂多糖刺激下体外培养奶牛乳腺上皮细胞活性和炎症相关基因表达的影响

本试验旨在研究苜蓿素对脂多糖(LPS)刺激下体外培养奶牛乳腺上皮细胞活性和炎症相关基因表达的影响。将体外培养的奶牛乳腺上皮细胞分成6组,对照组采用基础培养基,试验组在基础培养基中分别加入1μg/m L LPS(L组)、1μg/m L LPS和2.5μg/m L苜蓿素(L+2.5组)、1μg/m L LPS和5.0μg/m L苜蓿素(L+5组)、1μg/m L LPS和10.0μg/m L苜蓿素(L+10组)以及1μg/m L LPS和15.0μg/m L苜蓿素(L+15组),培养24 h。结果表明:1)与对照组相比,L组奶牛乳腺上皮细胞相对增殖率极显著降低(P0.01),而L+2.5组和L+10组则极显著升高(P0.01);2)与L组相比,L+10组和L+15组细胞超氧化物歧化酶活性显著升高(P0.05),而乳酸脱氢酶活性和一氧化氮浓度显著降低(P0.05);3)与L组相比,L+5组和L+10组细胞的白细胞介素8、白细胞介素1β、肿瘤坏死因子α、Toll样受体2和Toll样受体4的相对表达量均极显著降低(P0.01),L+5组细胞的髓样分化因子88相对表达量均显著降低(P0.05)。结果提示,在LPS刺激下,添加苜蓿素能够提高奶牛乳腺上皮细胞活性和抗氧化性能,抑制炎症相关基因的表达,发挥抗炎作用。     

围生期奶牛能量负平衡与免疫抑制的关系

为了探讨围生期奶牛能量负平衡与免疫抑制的关系,试验随机选取2个集约化牛场围生期奶牛各60头,以能量负平衡为判定标准,β-羟丁酸(BHBA) 1. 2 mmol/L、游离脂肪酸(NEFA)0. 4 mmol/L、葡萄糖(GLU)0. 3 mmol/L为发病组,BHBA 1. 2 mmol/L、NEFA 0. 4 mmol/L、GLU0. 3 mmol/L为对照组,对试验牛能量代谢指标、矿物质代谢指标、免疫功能指标进行组间独立样本t检验和Spearman相关性分析,并利用二元Logistic回归分析预测疾病,最后利用受试者工作特征曲线(ROC)分析确立分界值和诊断效果。结果表明:发病组奶牛血液中Ca、Mg、白细胞介素-1(IL-1)、白细胞介素-2(IL-2)、白细胞介素-6(IL-6)的水平和对照组奶牛相比差异极显著(P0. 01),发病组奶牛血液中C反应蛋白(CRP)、结合珠蛋白(HP)、肿瘤坏死因子(TNF-α)的水平和对照组奶牛相比差异显著(P0. 05),并且发病组奶牛血液中的Ca、Mg水平低于对照组,IL-1、IL-2、IL-6、HP、CRP、TNF-α的水平高于对照组; IL-1、IL-2、IL-6、HP、TNF-α水平与BHBA浓度呈显著或极显著正相关(P0. 05或P0. 01),Ca水平与GLU浓度呈极显著正相关(P0. 01),IL-2、HP水平与GLU浓度呈显著负相关(P0. 05),Ca、P、Mg水平与NEFA浓度呈显著或极显著负相关(P0. 05或P0. 01),IL-2、HP、TNF-α水平与NEFA浓度呈显著正相关(P 0. 05);经二元Logistic回归分析确定Ca、IL-1、IL-2、IL-6、HP、TNF-α水平6项指标与围生期泌乳奶牛免疫抑制相关;经ROC分析确立了HP、TNF-α2项免疫抑制指标的预警值为HP 29. 49 ng/L、TNF-α219. 90 ng/L。     

奶牛产后子宫炎早期预警指标确立与评估

为研究奶牛血清中代谢指标和激素指标与子宫炎的关系以及对子宫炎发生风险的预警作用,试验在新疆石河子地区某牛场选择产后1～10 d荷斯坦奶牛35头,根据临床症状、阴道分泌物性状、B超检查分为患病组(20头)、正常组(15头),检测子宫炎奶牛与正常奶牛血清中代谢指标和激素指标。结果表明:子宫炎奶牛血清中雌二醇(E2)、促卵泡生成素(FSH)的水平低于正常奶牛(P0.05),孕酮(P4)、前列腺素(PGE2)、门冬氨酸氨基转移酶(AST)含量高于正常奶牛(P0.05),其他指标各组之间差异不显著;通过对筛选出的指标E2、P4、FSH、PGE2、AST进行Pearson相关系数分析,发现P4与E2、FSH呈负相关,其中与FSH极显著相关,P4与AST、PGE2极显著正相关。根据接收者操作特征曲线(ROC)分析确定了该牛场子宫炎预警值为产后5～9 d奶牛血清中AST6.39 ng/mL,P49.53ng/mL,PGE23.92ng/mL。结果显示奶牛血清中P4、PGE2、AST指标超过预警值可作为子宫炎早期风险预测依据。     

同期发情处理的乏情奶牛血液病理学变化及其妊检阴性的风险预警

为了探究乏情奶牛同期发情处理前血浆生化指标的变化及对其输精后28 d妊检阴性(-)的风险预警意义,本试验在黑龙江省某集约化牛场选择了产后实施同期发情的奶牛共48头,根据试验奶牛产后40 d内是否发情分为发情组(E)23头和乏情组(A)25头,并对同期发情前试验牛进行血浆生化指标分析和输精后28 d妊检。结果显示:与发情奶牛相比,乏情奶牛血浆中孕酮(P_4)含量显著升高;雌二醇(E_2)、葡萄糖(Glu)、硒(Se)、牛组胺(HIS)、脑源性神经营养因子(BDNF)、牛组胺(HIS)含量显著降低。应用Pearson相关分析和二元Logistic回归模型确定了应用三针法前当奶牛血浆HIS含量 26.12μg/L,输精后28 d妊检为阴性(-)的风险增大;应用五针前当奶牛血浆中E_2含量46.45 pmol/L时,输精后28 d妊检为阴性(-)的风险增大。     

高产奶牛产后卵巢囊肿的血液临床病理学变化及其预警评估

选择黑龙江省某牛场产后60~90d奶牛共70头(平均泌乳量(34.55±8.44)kg/d,(2.24±1.01)胎次)。通过对发情和卵巢囊肿奶牛生殖激素,能量代谢,肝功和矿物质等10项指标检测,结合Pearson相关分析、二元Logistic分析以及ROC分析,结果显示:牛场卵巢囊肿发病率为9.8%,其主要原因为能量负平衡;卵巢囊肿奶牛血浆中牛促卵泡素(FSH)、牛促黄体激素(LH)和牛孕酮(P_4)的质量浓度均低于发情组,但仅牛雌二醇(E_2)质量浓度((101.95±6.2)ng/L)显著高于发情组((91.97±10.62)ng/L);牛场在14~21d和60~90d卵巢囊肿组与发情组相比游离脂肪酸(NEFA)、β-羟丁酸(BHBA)和门冬氨酸氨基转移酶(AST)均显著的升高,而Glu显著低于发情组;当产后14~21d奶牛血浆中BHBA>0.855 mmol/L、NEFA>0.585 mmol/L、AST>77.0U/L时,患卵巢囊肿的风险增高。结果表明:能量负平衡引起奶牛产后生殖激素分泌紊乱,进而造成卵泡囊肿。奶牛产后血浆NEFA、BHBA和AST等指标可作为卵巢囊肿发生的风险预警。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.     

Mechanism of Action of Probiotic Function of Lactobacilli

乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.     

Evaluation of serum concentrations of biomarkers of skeletal metabolism and results of radiography as indicators of severity of osteochondrosis in foals

OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.