利用血液蛋白及酶多态性分析技术鉴定山羊亲缘关系的研究

选择血红蛋白(Hb)、运铁蛋白(Tf)、亮氨酸胺肽酶(LAP)、前白蛋白3(Pa3)、淀粉酶(Amy)对可能为同卵双生波尔山羊的亲缘关系鉴定。结果发现，除淀粉酶(Amy)和亮氨酸胺肽酶(Lap)外，其余位点均有多态性。由蛋白位点的基因型判定羔1、羔2基因型完全一致，是供体后代的概率为75G，受体后代的概率为22．8％，从而确认羔1、羔2的双亲为供体公羊和供体母羊。     

马动脉炎病毒RT-PCR检测方法的建立

根据已报道的马动脉炎病毒基因组保守基因核苷酸序列，设计并合成了1对引物，通过对影响PCR扩增因素的筛选，成功地从病毒感染的细胞中扩增出约200bp的片段，与理论设计值(204bp)大小一致。而正常的RK-13、BHK-21和Vero细胞和同为动脉炎病毒科的猪繁殖与呼吸道综合征病毒(PRRSV)作为对照的扩增结果均为阴性。敏感性试验表明，该方法可以检测出10^-4个TCID50的病毒含量，说明具有较好的敏感性。     

肉仔鸡低血糖-尖峰死亡综合征的流行病学分析和病原初探

自2003年6月份以来，在我国北方肉鸡饲养基地普遍流行了一种主要侵害肉仔鸡的疾病，低血糖-尖峰死亡综合征(HSMS)。HSMS发病日龄集中于10～18日龄．临床表现为突然出现的高死亡率，病鸡头部震颤、运动失调、昏迷、失明、死亡。先后对黑龙江、吉林、辽宁、内蒙古、天津、河北、山东等省进行调查．发现HSMS在这几个省的很多肉鸡饲养区广泛存在。对发生该病的259个肉鸡饲养户的调查结果显示．该病与祖代种鸡没有关系．病原可能来自受污染的父母代种鸡场。平均发病率为17．9％。发病鸡群血糖浓度极显著的下降。从2个发病鸡群的病鸡肠道内容物中观察到病毒颗粒．病毒呈多形性．有囊膜，直径约80-100nm，表面有棒状纤突，长约10-20nm．与冠状病毒类似。     

抗鸡传染性腺胃炎酶标抗体的制备和应用

在前期从患病鸡腺胃内分离到呼肠孤病毒的基础上，以该病毒为免疫原对兔进行免疫制备高免血清，分离纯化IgG，进行辣根过氧化物酶（HRP）标记，并用ELISA双抗体夹心法对山东省青岛、枣庄、泰安等地区采集的传染性腺胃炎的腺胃样品进行检测，结果表明制备的抗鸡传染性腺胃炎呼肠孤病毒的酶标抗体检测传染性腺胃炎特异性强、灵敏度高、较简便。     

Noninferiority trial investigating the efficacy of a nonantibiotic intramammary therapy in the treatment of mild‐to‐moderate clinical mastitis in dairy cows with longer lasting udder diseases

A nonblinded, positively controlled, noninferiority trial was conducted to evaluate the efficacy of an alternative, nonantibiotic therapy with Masti Veyxym^® to reduce ineffective antibiotic usage in the treatment of nonsevere clinical mastitis (CM) in cows with longer lasting udder diseases. The solely intramammary treatment with Masti Veyxym^® (three applications, 12 hr apart) and the combined treatment with Masti Veyxym^® and antibiotics as usual on the farm according to label of the respective product were compared with the reference treatment of solely antibiotic therapy. The matched field study was conducted on eight free‐stall dairy farms located in Eastern Germany. Cases of mild‐to‐moderate CM in cows with longer lasting high somatic cell counts in preceding dairy herd improvement test days and with previous CM cases in current lactation were randomly allocated to one of the three treatment groups. A foremilk sample of the affected quarter was taken before treatment and again approximately 14 days and 21 days after the end of therapy for cyto‐bacteriological examination. Primary outcomes were clinical cure (CC) and no CM recurrence within 60 days after the end of treatment (no R60). Bacteriological cure (BC) and quarter somatic cell count (QSCC) cure were chosen as secondary outcomes although low probabilities of BC and QSCC cure for selected cows were expected. The study resulted in the following findings: the pathogens mostly cultured from pretreatment samples were Streptococcus uberis, followed by Staphylococcus aureus and coagulase‐negative staphylococci. There were no significant differences between the two test treatments in comparison with the reference treatment regarding all outcome variables. The sole therapy with Masti Veyxym^® resulted in a numerically lower likelihood of BC without significant differences to the reference treatment. The combined therapy group showed a numerically higher nonrecurrence rate than the two other treatment groups and noninferiority compared to the reference treatment was proven. Having regard to the selection criteria of cows in this study, the findings indicated that sole treatment with Masti Veyxym^® in nonsevere CM cases may constitute an alternative therapy to reduce antibiotics. However, noninferiority evaluations were mostly inconclusive. Further investigations with a larger sample size are required to confirm the results and to make a clear statement on noninferiority.     

Effects of concentrate‐to‐forage ratios and 2‐methylbutyrate supplementation on ruminal fermentation,bacteria abundance and urinary excretion of purine derivatives in Chinese Simmental steers

This study evaluated the effects of dietary concentrate levels and 2‐methylbutyrate (2MB ) supplementation on performance, ruminal fermentation, bacteria abundance, microbial enzyme activity and urinary excretion of purine derivatives (PD ) in steers. Eight ruminally cannulated Simmental steers (12 months of age; 389 ± 3.7 kg of body weight) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Moderate‐concentrate (400 g/kg diet [MC ]) or high‐concentrate (600 g/kg diet [HC ]) diets were fed with or without 2MB (0 g/day [2MB ?] or 15.0 g/day [2MB +]). Dry matter intake and average daily gain increased, but feed conversion ratio decreased with the HC diet or 2MB supplementation. Ruminal pH decreased, but total volatile fatty acid increased with the HC diet or 2MB supplementation. Molar proportion of acetate and acetate‐to‐propionate ratio decreased with the HC diet, but increased with 2MB supplementation. Propionate molar proportion and ruminal NH ₃‐N content increased with the HC diet, but decreased with 2MB supplementation. Neutral detergent fibre degradability decreased with the HC diet, but increased with 2MB supplementation. Crude protein degradability increased with the HC diet or 2MB supplementation. Abundance of Ruminococcus albus , Ruminococcus flavefaciens , Fibrobacter succinogenes and Bufyrivibrio fibrisolvens as well as activities of carboxymethyl cellulase, cellobiase, xylanase and pectinase decreased with the HC diet, but increased with 2MB supplementation. However, abundance of Prevotella ruminicola and Ruminobacter amylophilus as well as activities of α‐amylase and protease increased with the HC diet or 2MB supplementation. Total PD excretion also increased with the HC diet or 2MB supplementation. The results suggested that growth performance, ruminal fermentation, CP degradability and total PD excretion increased with increasing dietary concentrate level from 40% to 60% or 2MB supplementation. The observed diet × 2MB interaction indicated that supplementation of 2MB was more efficacious for improving growth performance, ruminal fermentation and total PD excretion with promoted ruminal bacteria abundance and enzyme activity in the MC diet than in the HC diet.     

Effect of gut stress induced by oxidized wheat gluten on the growth performance,gut morphology and oxidative states of broilers

This study was to investigate the effect of oxidized wheat gluten (OG) on growth performance, gut morphology and its oxidative states of broilers. One hundred and eighty‐day‐old male broilers (10 chicks/pen) were randomly allocated into three dietary treatments: control diet (CON), diet with 8% wheat gluten (WG) and diet with 8% OG with six pens/treatment. Body weight (BW) (21 and 35 days) and average daily gain (ADG) (1–21 days and 22–35 days) decreased (p < .05) and feed conversion ratio (FCR) (1–21 days and 22–35 days) increased (p < .05) in OG treatment. Feed intake (FI) decreased (p < .05) in WG and OG treatments during 22–35 days. However, FI was not influenced by dietary treatments during 1–21 days (p > .05). The OG‐fed broilers had a lower faecal pH value (p < .05) and higher faecal moisture content (p < 05) at 14, 21, 28 and 35 days. Villus height, crypt depth and V/C value were not different (p > .05) among treatments at 21 and 35 days. Lipid peroxidation (LPO) (21 and 35 days) and malondialdehyde (MDA) (35 days) content in crop of OG treatment increased (p < .05). Oxidized glutathione (GSSG) (21 days), LPO (21 and 35 days) and MDA (21 and 35 days) content in ileum of OG treatment increased (p < .05). The reduced glutathione/oxidized glutathione (GSH/GSSG) (21 days) and (GSH) (35 days) in ileum of OG treatment decreased (p < .05). The present findings indicate that OG might be a stressor for broiler gut, which could induce oxidative stress both in crop and in ileum, and the diarrhoea as well. The growth performance of broiler was consequently depressed.     

补饲水平对甘肃高山细毛羊生产性能的影响

为了探讨补饲水平对高寒牧区甘肃高山细毛羊生产性能的影响,本研究选择12月龄甘肃高山细毛后备母羊48只,随机分为补饲Ⅰ组(颗粒饲料)、Ⅱ组[玉米(Zea mays)和苜蓿(Medicago sativa)干草]、Ⅲ组[玉米、苜蓿干草和燕麦(Avena sativa)干草]和对照组(不补饲),进行冬春季"放牧"+"补饲"试验,比较了后备母羊体重,剪毛量,毛纤维长度、细度、白度、强度和伸度等物理指标。结果表明,试验Ⅰ组、Ⅱ组和Ⅲ组试验羊平均日增重分别比对照组高148.29%、133.09%、96.76%;剪毛量各组之间差异不显著(P0.05);自然长度和伸直长度对照组均极显著低于Ⅰ、Ⅱ、Ⅲ组(P0.01),且自然长度Ⅰ组极显著高于Ⅱ组和Ⅲ组(P0.01),伸直长度Ⅲ组极显著低于Ⅰ组和Ⅱ组(P0.01),Ⅱ组显著高于Ⅰ组(P0.05);细度(直径)Ⅰ组最大,Ⅰ组极显著高于对照组(P0.01),且显著高于Ⅱ组(P0.05);白度Ⅰ组最高,Ⅰ组显著高于对照组(P0.05),其他各组之间均差异不显著(P0.05);强度和伸度补饲组均极显著高于对照组(P0.01),且强度Ⅰ组极显著高于Ⅱ组(P0.01),伸度Ⅰ组显著高于Ⅱ组(P0.05)。由此可见,补饲对体重、羊毛品质都有显著正向影响,高寒牧区冬春季节牧草营养缺乏,为了满足羊只营养需要,使甘肃高山细毛羊后备母羊能在1.5岁进行配种繁殖,在冷季放牧的基础上补饲1号料为最佳选择。     

拮抗草坪蘑菇圈生防菌剂的关键发酵工艺及优化

为获得草坪蘑菇圈病原菌拮抗生防菌剂的优化参数,本研究以拮抗蘑菇圈病原-肉褐麟小伞菌(Lepiota brunneo-incarnata)的黄曲霉(Aspergillus flavus)CP01为目的菌,对其分别进行液、固态发酵条件的优化。通过单因素试验优化发酵时间、培养温度、培养基类型及初始pH等条件,对最佳液态发酵条件进行筛选;并通过单因素试验优化培养物料、外源养分、发酵时间和温度等条件,对最佳固态发酵条件进行筛选。结果表明:液体发酵条件下,马铃薯葡萄糖液体培养基(PDB)中菌丝体干重含量最高,而在青秸秆粉浸提液培养基(CSE)中有效菌数最高,且该发酵液对蘑菇圈病原的抑菌效果最佳,培养基最适初始pH为5、最佳发酵条件为30℃、5d;在固态发酵条件下,最适发酵培养物料为经浸提法去营养化的青秸秆粉,外源添加养分为料水比1∶50的青秸秆粉浸提液,最适发酵条件为33℃、9d。以上优化参数为草坪蘑菇圈生防菌剂的研发奠定了基础。     

紫花苜蓿9个盐响应相关基因表达模式

紫花苜蓿(Medicago sativa)是世界上种植面积最大、应用最广泛的豆科牧草。由于其耐盐性中等,其在我国北方地区的产量和种植受到土壤盐渍化的限制。因此提高紫花苜蓿的耐盐性具有重要的科学和生产意义。为此,以龙牧801紫花苜蓿(M.sativa‘Longmu 801’)为试验材料,采用实时荧光定量PCR技术分析了不同浓度NaCl胁迫下9个盐胁迫蛋白质组筛选出的盐响应相关基因的表达模式。结果显示,处理时间和处理浓度对9个基因的相对表达量均有显著性影响,表明这9个基因均在紫花苜蓿盐胁迫应答中发挥着一定作用。处理1h时,G6PI、ABP19a、Trx-h1、PR bet 1、FBPA、6PGDH和ALDH这7个基因的相对表达量在不同NaCl胁迫浓度处理的紫花苜蓿中均显著上调,RRM和GDPD在NaCl处理2h后开始上调。除G6PI基因,其他8个基因在0.4%NaCl处理下相对表达量显著高于0.2%和0.8%NaCl处理。这些基因参与糖代谢、信号转导和胁迫响应。以上结果表明,紫花苜蓿的耐盐性极其复杂,涉及到多基因的表达和代谢通路的调控。研究结果有助于全面研究并了解紫花苜蓿的盐响应相关基因的表达模式,促进紫花苜蓿耐盐分子育种。