荷斯坦奶牛妊娠早期血浆差异蛋白质组学研究

采用二维凝胶电泳(2-DE)结合MADIL-TOF-TOF串联质谱法对荷斯坦奶牛妊娠早期血浆进行蛋白质组学分析鉴定,发现了6种差异蛋白在妊娠后表达量上调,分别是凝集素1前体、转甲状腺素蛋白前体、3,5-二碘水杨酸络合的牛血清白蛋白的晶体结构的A链、白蛋白、未命名蛋白产物及结合珠蛋白-25同源物1前体。对这6种蛋白在不同时期含量的变化趋势及可能引起每种蛋白含量变化的原因和在妊娠过程中的功能进行了分析。     

采用HPLC法测定双峰驼血浆咪达唑仑浓度

旨在建立测定双峰驼血浆咪达唑仑浓度的高效液相色谱紫外检测法(HPLC-UV),为后续的特异性探针药物法研究双峰驼CYP3A酶体内活性提供方法学支撑。首先采用乙腈沉淀血浆样品蛋白,离心后经022μm有机针孔滤膜过滤,采用色谱柱为Sigma C18柱(46 mm×150 mm,5μm),流动相为乙腈:001 mol·L-1PBS缓冲溶液=60∶40(V/V),流速为05 mL·min~(-1),柱温为30℃,检测波长为254 nm等色谱条件进行样品检测。结果表明:咪达唑仑的保留时间为1324 min,峰形良好、无杂峰干扰,并在039~25μg·mL~(-1)浓度范围内线性关系良好(r2=0999,n=7)。高、中、低3个浓度的相对回收率与绝对回收率均符合检测要求。因此,本试验建立的测定双峰驼血浆咪达唑仑浓度的HPLC紫外检测方法灵敏度高,稳定性好,分离完全,峰形良好。     

反向高效液相色谱法检测双峰驼血浆中对乙酰氨基酚的含量

试验旨在建立测定双峰驼血浆中对乙酰氨基酚浓度的反相高效液相色谱法（reversed phase high-performance liquid chromatography,RP-HPLC）。选用Supelco Discovery C18色谱柱（150 mm×4.6 mm,5 μm）,以甲醇：水=20：80为流动相,流速为0.5 mL/min,紫外检测波长为248 nm,采用甲醇直接沉淀蛋白方法处理血浆样品。结果表明,对乙酰氨基酚的保留时间为6.5 min,分离度良好,无血浆杂峰干扰,且在0.08~10.24 μg/mL浓度范围内线性关系良好（R²=0.9997,n=8）,检测限为0.10 μg/mL,日内和日间精密度RSD均小于7.0%,绝对回收率为80.39%~93.56%,相对回收率为94.66%~104.73%。因此,本试验所建立的RP-HPLC法操作简便、准确,重复性好,专属性强,适用于检测双峰驼血浆中对乙酰氨基酚浓度,为后续的借助特异性探针药物--对乙酰氨基酚研究双峰驼CYP1A酶的体内活性提供了可靠而准确的定量方法。     

双峰驼适应荒漠草原的特性

双峰驼适应进化的特性表现在多个方面,经过恶劣环境的自然选择所形成的双峰驼,其遗传资源无疑具有特别的珍贵性,由于人为和自然的因素,双峰驼的数量在急剧减少。对于双峰驼适应环境的具体表现和生理机制有一个较为全面的了解是必要的。笔者综述了该方面的研究进展,以期对双峰驼保护和利用工作有所帮助。     

荷斯坦奶牛产后期血浆差异蛋白质组学研究

为了检测和分析荷斯坦奶牛分娩后不同时期血浆中的差异蛋白。采用二维凝胶电泳(2-DE)分离血浆蛋白,考马斯亮蓝染色,PDQuest8.0软件检测分析差异蛋白点,MADIL-TOF-TOF串联质谱鉴定差异蛋白点。结果发现3种差异蛋白,分别是结合珠蛋白、免疫球蛋白γ2链C区和3,5-二碘水杨酸络合的牛血清白蛋白。对3种蛋白在产后不同时期的趋势进行分析,可能分别与产后应激、免疫应答和物质运输有关。荷斯坦奶牛产后不同时期血浆中蛋白表达存在差异,对差异表达蛋白的研究有利于探索奶牛产后生理机制,可作为产后饲养、配种及防止产后感染等研究的基础。     

不同环境温度下的育雏效果

不同环境温度下的育雏效果＠张永江＠陈万中￥吉林省农科院畜牧分院不同环境温度下的育雏效果张永江陈万中（吉林省农科院畜牧分院怀德１３６１００）目前各养鸡场家和专业户采用不同的给温方法进行雏鸡培育工作,所取得的结果也各有不同,本文就常规低温、高温和超低温育雏谈一...     

双峰驼在不同饮水条件下血清中促甲状腺激素三碘甲腺原氨酸和甲…

双峰驼血清中促甲状腺和甲状腺在不同饮水条件下的含量均是冬季高于夏季。     

阿拉善驼妊娠前后卵巢差异蛋白质组学分析

为了比较阿拉善双峰驼妊娠前后卵巢中差异表达的蛋白质,采集非繁殖季节未妊娠和妊娠期阿拉善驼卵巢,运用双向电泳技术构建双峰驼卵巢蛋白质凝胶电泳图谱,凝胶经考马斯亮蓝染色后,用PDQuest 8.0软件检测3倍以上的表达变化的蛋白点。结果表明在两组样品中共检测到10个差异蛋白点,这些蛋白点经MALDI-TOF/TOF-MS鉴定为8种蛋白质,包括谷胱甘肽S-转移酶、过氧化物酶-6、丙酮酸激酶同工酶M1/M2异构体1、纤维蛋白原β链前体、核纤层蛋白A/C转录物变异体1、视黄醛脱氢酶1和一种未命名的蛋白质。其中核纤层蛋白A/C转录物变异体1仅在未妊娠卵巢中表达,视黄醛脱氢酶1在双峰驼妊娠卵巢组织中表达量下降,而其他蛋白质在妊娠卵巢中表达量增加。这些差异表达蛋白质的发现为了解双峰驼妊娠期卵巢的生理机制奠定了基础。     

基于转录组测序筛选高盐培养下双峰驼肾皮质细胞差异表达基因

为探讨双峰驼的高盐适应机制并筛选出受高盐调控的基因,本试验采用RNA-Seq技术对双峰驼肾皮质细胞进行转录组测序分析。试验分为2个组,等渗培养基处理的肾皮质细胞为对照组（Control）,高渗培养基处理的肾皮质细胞为高渗组（HS）,进行转录组测序,从而筛选出一系列差异表达基因（DEGs）,并用GO富集和KEGG富集分析对DEGs进行基因功能和途径的注释。结果显示,在高盐环境下双峰驼肾皮质细胞中共有4 854个显著DEGs;GO富集分析显示,DEGs显著富集在G蛋白偶联受体活性、受体结合、核酸结合转录因子活性、质膜、离子通道的活动和免疫反应等功能中;KEGG通路富集显示,DEGs显著富集在信号传导、辅助因子和维生素代谢、信号分子相互作用、运输和分解代谢、免疫系统等通路中;采用实时荧光定量PCR技术随机检测了参与高盐代谢的3个差异表达基因,其表达趋势与RNA-Seq一致,可以验证RNA-Seq技术的准确性。因此,双峰驼肾皮质细胞的高盐耐受性可能与这些途径和通路中重要基因的调控有关。本试验结果将为进一步揭示双峰驼耐盐性的分子调控机制提供重要的参考依据。     

双峰驼初乳IgG的分离纯化及其向幼驼的转移

通过跟踪检测内蒙古阿拉善双峰母驼初乳IgG和幼驼血清IgG的动态含量，探讨了影响幼驼由母体获得被动免疫的因素。采用硫酸铵沉淀法2次沉淀乳清蛋白，再结合离子交换层析方法，从双峰驼初乳中分离纯化驼IgG；再用SDS—PAGE鉴定其纯度，测定其分子质量；用分离纯化的驼IgG免疫家兔，制备效价至少为1：32的兔抗驼IgG抗血清，采用单向免疫扩散法测定双峰驼初乳和幼驼血清IgG含量。揭示了初乳IgG向幼驼血液的转移模式，为生产中更好地饲养幼驼提供科学依据。     

双峰驼肾近曲小管和远曲小管的体视学研究

应用光镜和电镜技术,测算了双峰驼肾近曲小管和远曲小管微细结构的体视学参数,并与黄牛进行了比较.结果显示,双峰驼肾近曲小管刷状缘体积、微绒毛表面积、吞饮小泡和溶酶体体积均显著大于黄牛;远曲小管上皮细胞的线粒体体密度、侧基底面质膜面密度和周界长度也明显大于黄牛.这些结构的差异表明,双峰驼肾脏重吸收水分的能力更强大.     

The gross anatomy of the cranial cervical ganglion and its branches in the bactrian camel (Camelus bactrianus)

Ten specimens of the head and neck of the Bactrian camel (Camelus bactrianus) were dissected to study the situation, arrangement and branches of the cranial cervical ganglion (ganglion cervicale craniale). The ganglion was a greyish fusiform structure, averaging 15–20 mm in length, 4–6 mm in width and 3 mm in thickness, located on the rostro-lateral surface of the longus capitis and covered by the mandibular gland. The branches of the cranial cervical ganglion included the internal carotid nerve, external carotid nerve, jugular nerve and the branches connecting with the glossopharyngeal, vagus, hypoglossal and first cervical nerves.     

双峰驼奶对2型糖尿病大鼠肾组织结构及其细胞凋亡的影响

观察双峰驼奶对2型糖尿病大鼠肾组织结构及其细胞凋亡的影响,为临床预防和治疗糖尿病及其并发症提供理论参考。通过链脲佐菌素（streptozotocin,STZ）结合高糖高脂饮食诱导构建2型糖尿病（type 2diabetes mellitus,T2DM）大鼠模型,糖尿病大鼠随机分为糖尿病模型组（diabetes model control,DMC）、低剂量骆驼奶组（2ml/d）（low camel milk,LCM）、高剂量骆驼奶组（5ml/d）（high camel milk,HCM）和盐酸二甲双胍组（200mg/kg）（metformin hydrochloride,MTH）,每组12只,另设正常对照组（control group,C）。建模成功后,试验组处理4周后检测大鼠空腹血糖（FPG）浓度变化;采用苏木精-伊红（HE）染色方法检查肾脏组织的病理变化;采用荧光定量Real time-PCR法检测Bax（Bcl2-associated X protein,Bax）、Bcl-2（B cell lymphoma/lewkmia-2,Bcl-2）、细胞凋亡蛋白酶3（cysteinyl aspartate specific proteinase 3,Caspase 3）和核转录因子（nuclear factor-kappa B,NF-κB）mRNA的表达丰度;采用免疫组织化学染色法和免疫印迹法观察肾脏组织凋亡相关蛋白Bax和Bcl-2的表达变化。结果显示,与DMC组相比,LCM、HCM、MTH组大鼠FPG浓度均显著下降（P〈0.01）,LCM组大鼠胰岛素敏感性指数（ISI）显著升高（P〈0.01）;HE染色显示DMC组肾小球体积增大,囊腔变小,肾小球内出血严重,而LCM、HCM、MTH组肾小球体积、囊腔间隙较正常,肾小球出血明显减少;Bax mRNA和NF-κB mRNA在DMC组中表达量最高,Bcl-2mRNA和Caspase 3mRNA分别在LCM组和HCM组有较高的表达量;与DMC组相比,LCM、HCM、MTH组大鼠肾脏细胞Bax蛋白表达减低,Bcl-2蛋白表达升高。结果表明,双峰驼奶能显著降低糖尿病大鼠肾脏损伤,抑制其细胞凋亡,对糖尿病及其并发症的治疗发挥有效作用。     

Screening for reproductive biomarkers in Bactrian camel via iTRAQ analysis of proteomes

Bactrian camel is an ancient and precious species of livestock; that is, unique resources exist in the desert and have important economic and scientific value. In recent years, the number of Bactrian camels has declined sharply. Due to its long reproductive cycle and seasonal oestrus, the mechanism of oestrus is unknown. To identify candidate biomarkers of reproduction, we performed a comprehensive proteomic analysis of serum from Bactrian camel in oestrus and non-oestrus, using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with tandem mass spectrometry. We identified 359 proteins, of which 32 were differentially expressed: 11 were up-regulated and 21 were down-regulated in samples from camels in oestrus. We validated the differential expression of a subset of these proteins using qPCR and Western blot. Gene ontology annotation identified that the differentially expressed proteins function in cellular processes, metabolic processes and immune system processes. Notably, five of the differentially expressed proteins, PCGF5, histone H1.2, RBP4, FOLR1 and ANTXR2, are involved in reproductive regulatory processes in other animals. KEGG enrichment analysis demonstrated significant enrichment in several cardiac-related pathways, such as ‘dilated cardiomyopathy’, ‘hypertrophic cardiomyopathy’, ‘cardiac muscle contraction’ and ‘adrenergic signalling in cardiomyopathy’. Our results suggest that candidate biomarker (PCGF5, histone H1.2, RBP4, FOLR1 and ANTXR2) discovery can aid in understanding reproduction in Bactrian camels. We conclude that the profiling of serum proteomes, followed by the measurement of selected proteins using more targeted methods, offers a promising approach for studying mechanisms of oestrus.     

Effects of high ambient temperature on plasma metabolomic profiles in chicks

Exposure to high ambient temperature is detrimental to the poultry industry. To understand the influence from a metabolic perspective, we investigated the effects of exposure to high ambient temperature on plasma low‐molecular‐weight metabolite levels in chicks using gas chromatography/mass spectrometry‐based non‐targeted metabolomic analysis. Heat exposure for 4 days suppressed growth and food intake. Of the 92 metabolites identified, the levels of 29 decreased, whereas the levels of nine increased. We performed an enrichment analysis on the identified metabolites and found 35 candidates for metabolic processes affected by heat exposure. Among them, the sulfur amino acid metabolic pathway was clearly detected and the levels of the following related metabolites were decreased: cystathionine, cysteine, cystine, homocysteine and hypotaurine. Changes in the kynurenine pathway in tryptophan metabolism, which is linked to the immune system and oxidative stress, were also observed: kynurenine and quinolinic acid levels increased, whereas nicotinamide levels decreased. These results suggest the possible involvement of various metabolic processes in heat‐exposed chicks. Some of these metabolites would be important to understand the mechanism of biological responses to high ambient temperature in chicks.     

The detection of subclinical mastitis in the bactrian camel (Camelus bactrianus) by somatic cell count and California mastitis test

Milk samples (n=160) from 7 clinically healthy bactrian camels were cultured to detect subclinical udder infection. The samples were assessed by the Californian mastitis test (CMT) and somatic cell count (SCC). Bacteria were recovered from 36 (22.5%) of the milk samples. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were the main organisms found.Infected quarters had significantly higher mean values for the SCC (p<0.01) and CMT (p<0.001) than non-infected quarters. All 7 camels were infected with CNS but only 4 with S. aureus. CMT values for S. aureus-infected camels were significantly higher than for those only infected with CNS. The values for SCC and CMT were significantly influenced by the stage of lactation (p<0.05). No significant difference was found from the effect of the quarters. Both SCC and CMT were of value in predicting the infection status of the udder.Abbreviations CMT California mastitis test - SCC somatic cell count - CNS coagulase-negative staphylococci     

Effects of high ambient temperature and restricted feed intake on nitrogen utilization for milk production in lactating Holstein cows

Three experiments were performed to examine the effects of high ambient temperature and feed restriction on nitrogen (N) utilization in lactating cows. Experiment 1 investigated N utilization in four cows fed ad libitum in a 2 × 2 crossover design under constant moderate (18°C) or high (28°C) ambient temperatures. The milk N secretion (P < 0.01) and protein concentration (P < 0.05) decreased under high ambient temperature. Experiment 2 investigated N utilization in four cows under constant moderate ambient temperature in a 2 × 2 crossover design with ad libitum or 70% ad libitum feed intake. The milk N secretion and protein concentration both decreased with feed restriction (P < 0.05). Experiment 3 investigated N utilization in four cows fed 70% ad libitum in a 2 × 2 crossover design under constant moderate or high ambient temperatures. The milk protein concentration decreased under high ambient temperature (P < 0.01). The milk N secretion tended to decrease under high ambient temperature (P < 0.10). Therefore, decreased N utilization for milk production at high ambient temperature is mainly caused by a reduced feed intake and the high ambient temperature itself.     

西藏黄牛不同海拔群体血清GCK、CK-MB、LDH酶含量比较

选择不同海拔高度（2 900～3 100,3 700～3 800,3 900～4 100,4 500～4 600m）饲养的西藏黄牛,用ELISA法测定不同海拔高度下牛血清中GCK、CK-MB、LDH酶的含量。采血后2h内对血清酶含量进行实地测定,并对结果进行统计分析。结果表明,GCK随海拔高度的升高,酶含量明显降低,呈现负相关性,组间方差分析为F=92.636,P=0.000,差异极显著;CK-MB含量亦随海拔高度的升高表现出显著性降低,呈现负相关性,组间方差分析结果为F=39.238,P=0.000,差异极显著;LDH酶系的含量变化幅度不大,组间方差分析结果F=3.223,P=0.029,差异显著。这表明高海拔对血清GCK、CK-MB酶（系）的含量具有一定的影响。     

Evaluation of feeding spray-dried bovine plasma protein on production performance of laying hens exposed to high ambient temperatures

The purpose of this study was to evaluate feeding 2 levels of spray-dried bovine plasma protein (SDP) on production performance of laying hens subjected to high ambient temperatures. Two groups of 96 Hy-Line W-98 hens (38 wk of age) were housed in each of 2 environmentally controlled chambers. At 40 wk of age, all hens were fed 3 diet treatments consisting of (1) a control diet (0% SDP); (2) the control diet supplemented with 0.75% SDP; and (3) the control diet supplemented with 1.50% SDP. Hens in each chamber (8 cages of 4 hens per cage) were ad libitum fed 1 of each diet for 5 wk. The heat stress (HS) chamber was maintained at 21°C (wk 1), 29°C (wk 2), and 35°C (wk 3 to 5). The thermoneutral chamber was maintained at 21°C during wk 1 to 5. A significant main effect of week was observed for hens maintained in the HS chamber for egg production, egg weight, egg mass, and feed consumption, which resulted in acute heat stress causing a reduction in these parameters. Hens fed the 1.50% SDP diet in the HS chamber produced greater (P < 0.05) egg mass on average than hens fed the control or 0.75% SDP diet (wk 1 to 5). During the second week of acute HS (wk 4), hens fed the control and 1.50% SDP diets had greater (P < 0.05) egg production than those fed the 0.75% SDP diet. During wk 5, hens in the HS chamber that were fed the 1.50% SDP diet produced more (P < 0.05) eggs than those fed the control diet. Therefore, based on the results of this study, acute HS negatively affected short-term production performance. In addition, feeding hens an SDP-supplemented diet may have a slight positive effect on production performance when maintained in acute HS conditions.