湖羊羔羊早期断奶前后瘤胃发酵与微生物区系的变化

本试验旨在研究湖羊羔羊早期断奶前后的瘤胃发酵与微生物区系变化,为幼龄反刍动物瘤胃发育变化理论以及羔羊的早期培育等提供依据。选择初生重接近的湖羊公羔(3.81 kg±0.55 kg)16只,1~7日龄饲喂母乳,8日龄与母羊分离,开始饲喂代乳粉(按8日龄体重的2%,分3次等量饲喂)和开食料(自由采食),35日龄断奶。分别于断奶前(21日龄)、后(42日龄)各选择5只羔羊进行屠宰,采集瘤胃内容物,测定瘤胃发酵、酶活和微生物区系。结果表明,断奶后羔羊的瘤胃总挥发性脂肪酸、乙酸、丙酸、丁酸及纤维素酶和α-淀粉酶活性均极显著高于断奶前(P<0.01)。断奶后瘤胃菌群多样性和丰富度均低于断奶前(P<0.05)。断奶前后的优势菌门均为拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes),其中,拟杆菌门为第一优势菌门,在断奶前后分别占61.96%和65.36%,厚壁菌门为第二优势菌门,在断奶前后分别占32.08%和24.03%;两菌门之和在断奶前后分别占瘤胃总菌门的94.04%和89.39%。断奶前后的优势菌属均为unidentified_Prevotellaceae,分别占21.85%和38.49%。断奶前后羔羊瘤胃微生物的功能没有显著变化,都主要集中在复制和修复、碳水化合物代谢和翻译等途径。以上结果说明,羔羊早期断奶后的瘤胃发酵和酶活增强,菌群多样性和丰富度有所降低,在早期断奶前后的优势菌群和功能相似。     

Effects of perioperative saphenous and sciatic nerve blocks,lumbosacral epidural or morphine–lidocaine–ketamine infusion on postoperative pain and sedation in dogs undergoing tibial plateau leveling osteotomy

ObjectiveTo compare the quality of postoperative analgesia and sedation after preoperative saphenous and sciatic nerve blockade, preoperative lumbosacral epidural injection and perioperative intravenous (IV) morphine, lidocaine and ketamine infusions in dogs undergoing stifle arthroscopy and tibial plateau leveling osteotomy (TPLO) under general anesthesia.Study designProspective, blinded, randomized, clinical comparison study.AnimalsA total of 45 dogs weighing 33.9 (15.9–56.7) kg and aged 5.2 (1.0–12.0) years, mean (range), undergoing elective unilateral TPLO for spontaneous cranial cruciate ligament rupture.MethodsClient-owned dogs were enrolled. Dogs were randomly assigned to one of three groups: group MLK, perioperative IV morphine, lidocaine and ketamine infusion; group EPID, lumbosacral epidural with ropivacaine and morphine; or group SSNB, saphenous and sciatic nerve blockade with ropivacaine. Routine stifle arthroscopy followed by TPLO surgery was performed. Sedation and pain scores were assessed at 0, 2, 4, 8 and 24 hours following extubation. Rescue analgesia was administered as prescribed by Glasgow composite pain score–short form score >5.ResultsSedation scores for MLK were higher than EPID and SSNB. Pain scores for SSNB were lower than those for EPID and MLK. No significant differences were found in anesthesia duration or surgery duration among groups. No dogs required rescue analgesia.Conclusions and clinical relevanceAlthough analgesia was adequate in all groups, the best combination of analgesia without increased sedation was recorded for SSNB.     

我国不同奶业产区规模化牧场饲料结构分析

在奶牛养殖规模化水平不断提高,养殖成本日益增加,且区域间差异明显的背景下,饲料作为奶牛养殖主要投入品,了解其在规模化牧场生产成本中的结构对于优化奶牛日粮结构、降低养殖成本、缩小区域间养殖效益差距等具有重要意义。本文利用2018—2020年规模牧场月度调研数据,分析了我国规模化牧场饲料结构情况。研究发现,大城市周边产区和南方产区的精饲料和粗饲料的比例相对更加科学,华北产区仍有较大提升空间,并且玉米、豆粕、玉米青贮和苜蓿草在日粮中占有较大比重,达到84.62%;不同产区奶牛日粮结构具有明显差距,玉米青贮和苜蓿草在西北产区占比最高,玉米和豆粕在南方产区占比最高。注重发挥区域优势,加大优质粗饲料和可替代饲料的利用,有利于降低养殖成本,提高养殖的科学性和可持续性。     

羊源D型多杀性巴氏杆菌感染小鼠脾脏转录组分析

试验旨在探索羊源D型多杀性巴氏杆菌入侵宿主脾脏组织中引发的免疫应答途径。首先利用羊源D型多杀性巴氏杆菌(HN01菌株)感染小鼠,建立羊源D型多杀性巴氏杆菌感染的动物模型;之后利用转录组测序技术获得感染小鼠与正常小鼠的脾脏转录组数据,并使用COG、KOG、eggNOG、GO、KEGG数据库对测序结果中的差异表达基因(differentially expressed genes,DEGs)进行功能注释与分析,同时对于显著富集到关键免疫通路的差异表达基因使用STRING软件和KEGG mapper进行蛋白互作分析,筛选出核心通路中起关键作用的基因;最后选取关键的10个基因进行实时荧光定量RT-PCR验证。转录组分析结果显示,与正常组相比,感染组中筛选出3 380个差异表达基因(P<0.01,log₂|FoldChange|≥0.5),其中1 691个基因上调,1 689个下调。基因功能富集分析结果表明,感染组脾脏中的差异表达基因主要发挥信号转导的功能,其主要参与的生物途径包括细胞因子与细胞因子受体互作通路、趋化因子信号通路、HIF-1信号通路、TNF信号通路。蛋白互作分析筛选出约28个核心差异表达基因,结合实时荧光定量RT-PCR验证后,其中9个基因的表达结果与测序一致,分别是C3、Cd4、Cxcl13、Lck、Gnai1、Grap2、IL-6、Cxcr6及Serping1基因。本研究初步证明了脾脏在抵抗多杀性巴氏杆菌入侵中参与了一系列免疫应答反应,为进一步研究羊源D型多杀性巴氏杆菌与宿主之间相互作用的分子机制奠定理论基础。     

哺乳动物卵母细胞非整倍体产生机制的研究进展

雌性生殖细胞进行减数分裂时易发生染色体分离错误而产生非整倍体卵母细胞,其受精后会产生非整倍体胚胎,导致出生缺陷或胚胎致死,是影响哺乳动物繁殖的重要因素。卵母细胞在第一次减数分裂前期发生同源染色体联会,此时DNA双链断裂引发重组。重组时缺乏交叉、重组事件数量的减少及交叉靠近端粒或着丝粒导致染色体发生同向分离或不分离,从而产生非整倍体卵母细胞。减数分裂期间,当染色体的端粒共向于同一极或没有完全附着在纺锤体微管上时,纺锤体组装检查点（spindle assembly checkpoint,SAC）被激活,E3泛素连接酶APC/Cyclome （APC/C）沉默,保护分离酶抑制蛋白（securin）和细胞周期蛋白B （cyclin B）不被降解,从而抑制分离酶和染色体的分离。直到所有染色体与纺锤体实现稳定的双极定向并正确排列到赤道板上,SAC关闭,染色体正确分离。卵母细胞中SAC蛋白缺失,导致SAC不能有效地监测端粒在纺锤体上的正确附着,发生染色体分离错误,从而产生非整倍体卵母细胞。因此,通过现代分子技术手段解析非整倍体卵母细胞所涉及的机制是保护哺乳动物生育的重要目标。作者主要介绍了卵母细胞减数分裂的特点,详细阐述了卵母细胞非整倍体发生的染色体分离错误的分子机制,以期为开发卵母细胞非整倍体的治疗手段提供参考。     

拷贝数变异在畜禽中的研究进展

拷贝数变异（copy number variation,CNV）具有多种形式的变异结构,在品种多样性、生物进化和疾病相关性等研究中起着重要作用,并具有片段长度大、覆盖范围广等特点。随着分子生物学的发展及DNA测序技术的日渐成熟,人们对遗传变异的研究不断向DNA分子水平深入,多态性标记在畜禽育种中已逐渐成为动物育种研究的趋势和主流。由于CNV对基因的调控和表达所造成的影响更为显著,因此,CNV在重要畜禽中的研究越来越多。目前,已检测出大量有关畜禽重要经济性状的基因序列变异,并有许多研究均表明CNV与动物的重要经济特征及疾病的发生有关。笔者主要通过参考国内外相关的研究报道,简述了CNV的相关研究背景、概念、突变机制,归纳总结了CNV对牛、羊、猪、鸡的经济性状、繁殖性状和疾病调控的影响,以期通过对这些重要畜禽的基因组学研究揭示其适应性遗传机理和表型性状差异的遗传基础,开发相应的分子遗传标记,为畜禽的标记辅助选育提供理论基础。     

肌肉发生相关长链非编码RNA研究进展

肌肉参与机体的运动、协调和体温调节等一系列生命活动,同时畜禽肌肉是人类重要的蛋白质来源。肌肉发生过程中的不同调控机制可引起肌肉发育的阶段性差异,而整个肌肉发育主要以两个时期（胚胎期和出生后）的发育状态体现。长链非编码RNA（lncRNA）是一类不具有蛋白编码能力且长度>200 nt的RNA分子。近年来,随着基因组学和分子生物学技术的高速发展,发现lncRNA广泛参与到肌肉发育的各个阶段,以多种作用机制调控肌肉的发育过程。作者介绍了肌肉的发育过程,综述了目前发现的与肌肉发育相关的lncRNAs及其作用机制,并阐述其在肌肉发育的不同阶段发挥的重要作用,为进一步研究肌肉发育相关lncRNAs提供了参考。     

人工养殖西伯利亚狍肠道微生物组成的雌雄差异

肠道微生物是一个复杂的生态系统,与宿主的健康、疾病以及其他因素有着密切的关系。目前,关于肠道菌群结构和功能的研究较多,但是对西伯利亚狍肠道菌群的研究还未见报道。西伯利亚狍(Capreolus pygargus)属于反刍动物,有独特的消化特性和微生物群来帮助它们适应高纤维的食物。本研究利用高通量测序技术在Illumina MiSeq平台上对14只狍肠道微生物进行16S rRNA基因分析。结果表明:个体样本中门类组成的差异,雌性个体JY3和雄性个体JY9除Firmicutes,Bacteroidetes,Proteobacteria这3类菌群外,其他门类菌群所占比例都很低。在科水平上,雌性个体中Ruminococcaceae在JY7和JY13肠道内含量相对较少,Bacteroidaceae在JY7,JY10和JY13中含量较高。雄性个体中Ruminococcaceae在JY11和JY14肠道内含量较少,Prevotellaceae在JY4中含量较少。在属水平上,雌性个体特有的菌属为Roseburia,Alloprevotella,Lachnospiraceae_incertae_sedis和Coprococcus;雄性个体特有的菌属为Fusobacterium,Succiniclasticum,Streptococcus和Clostridium sensu stricto。Alpha多样性分析表明,Ace指数范围2128.93—772.98,Chao1指数范围2128.89—785.27,Shannon指数范围5.44—3.78,Simpson指数范围0.07—0.01。在不同物种分类水平上,雌雄狍肠道内的优势菌群有些不同。PCoA(principal co-ordinates analysis)分析显示,狍样本几乎混在一起,没有明显差异。     

Residue Detection of Tetracyclines in Eggs by High Performance Liquid Chromatography

The study was aimed to establish the method of simultaneous detection of four kinds of tetracyclines,including oxytraoycline,tetracycline,chlortetracycline and doxyeycline in eggs by high performance liquid chromatography (HPLC).The eggs samples were detected with HPLC after through the procedure of extraction of 0.1 mol/L Na₂EDTA-Mcllvaine buffered solution,purification of Oasis MAX solid-phase extraction column,gradient elution of taking acetonitrile-0.01 mol/mL trifluoroacetic acid as mobile phase,separation with reverse liquid chromatography.The results showed that the calibration curves of four kinds of tetracyclines were linear correlated in the concentration range of 30 to 1 000 ng/g which the limits of detection was 10 ng/g and the limit of quantification was 50 ng/g.The average recoveries were 67.3% to 81.4%,relative standard deviations of intraday was 2.7% to 14.3%,relative standard deviations of inter-day was 0.9% to 5.0%.This mothod established in this study could simultaneously detect oxytetracycline,tetracycline,chlortetracycline and doxycycline drug residues in egg and it had high sensitivity,accuracy and precision.The method was rapid and simple to operate.     

Effect of Weaning Stress on Blood Physiology and Biochemistry Indexes and Immune Function in Dairy Buffalo Calves

To study the effect of weaning stress on dairy buffalo calves,5 healthy calves were chosen and weaned,and the blood samples were collected at 1 day before weaning,and 1,14 and 21 days after weaning to analysis blood physiology and biochemistry indexes and immune function.The results showed that the count of white blood cell,lymphocyte,erythrocyte,monocytes and hematocrit,aspartate aminotransferase activity,blood glucose,cortisol and immunoglobulin G contents of dairy buffalo calves were no significant difference between before and after weaning treatments (P > 0.05).The count of neutrophils at 21 d after weaning were significantly lower than that of 1 d before weaning of dairy buffalo calves (P < 0.05).However,lactate dehydrogenase and alanine aminotransferase activity in weaned treatments were significantly higher than those of animals in the treatment before weaning (P < 0.05).The serum urea nitrogen of dairy buffalo calves at 21 d after weaning was significantly higher than that of 1 d before weaning (P < 0.05) and hemoglobin content at 21 d after weaning was significantly lower than that at 1 day before weaning (P < 0.05).It indicated that the physiological and immunity function of dairy buffalo calves were challenged under weaning stress.