Study on Expression and Preliminary Immunocompetence of CIC Protein Containing Molecular Adjuvants     

ZHANG Shao-duo  LAI Yi-jun  JIANG Hui  CHENG Xin-ran  YE Yu-ting  HE Xi-yu  XU Le  SONG Bai-fen  YU Li-quan  CUI Yu-dong  MA Jin-zhu 《中国畜牧兽医》2017,44(12):3612-3617

To express CTB-IsdBid-Clfais(CIC) protein and evaluate its immunogenicity, CTB (as a molecular adjuvant) could be tandem linked with IsdBid-Clfais gene by the overlapping PCR method, then CTB-IsdBid-Clfais(CIC) was inserted into pET-32a(+) vector to construct recombinant plasmids pET-32a(+)-CTB-IsdBid-Clfais. The recombinant plasmids pET-32a(+)-CTB-IsdBid-Clfais were transformed into Escherichia coli (E.coli) BL21 to express the CTB-IsdBid-Clfa(CIC) protein, the CIC expression protein and its immune activity were detected by Western blotting and ELISA,respectively. The results showed that the length of CIC gene were 2 072 bp, and CIC was correctly inserted into the pET-32a(+) plasmids, the pET-32a(+)-CTB-IsdBid-Clfais recombinant plasmids were successfully constructed. Western blotting result confirmed that the molecular weight of CIC proteins was 95.9 ku, which were correctly expressed by E.coli BL21 with pET-32a (+)-CTB-IsdBid-Clfais plasmids. ELISA results showed that there was no significant difference among the CIC, IsdBid and Clfais protein groups (P>0.05), and there was extremely significant difference between CIC and BSA protein groups (P<0.01). In conclusion, the pET-32a(+)-CTB-IsdBid-Clfais recombinant plasmids were successfully constructed, CIC proteins were correctly expressed, and were able to react with serum from mice immunized with IsdBid and Clfais,respectively,therefore, CIC proteins had strong immune activity.  相似文献   

Effect of feed restriction on dairy cow milk production: a review     

Antoine Leduc  Sylvain Souchet  Marine Gel  Fabienne Le Provost  Marion Boutinaud 《Journal of animal science》2021,99(7)

In the dairy cow, negative energy balance affects milk yield and composition as well as animal health. Studying the effects of negative energy balance on dairy cow milk production is thus essential. Feed restriction (FR) experiments attempting to reproduce negative energy balance by reducing the quantity or quality of the diet were conducted in order to better describe the animal physiology changes. The study of FR is also of interest since with climate change issues, cows may be increasingly faced with periods of drought leading to a shortage of forages. The aim of this article is to review the effects of FR during lactation in dairy cows to obtain a better understanding of metabolism changes and how it affects mammary gland activity and milk production and composition. A total of 41 papers studying FR in lactating cows were used to investigate physiological changes induced by these protocols. FR protocols affect the entire animal metabolism as indicated by changes in blood metabolites such as a decrease in glucose concentration and an increase in non-esterified fatty acid or β-hydroxybutyrate concentrations; hormonal regulations such as a decrease in insulin and insulin-like growth factor I or an increase in growth hormone concentrations. These variations indicated a mobilization of body reserve in most studies. FR also affects mammary gland activity through changes in gene expression and could affect mammary cell turnover through cell apoptosis, cell proliferation, and exfoliation of mammary epithelial cells into milk. Because of modifications of the mammary gland and general metabolism, FR decreases milk production and can affect milk composition with decreased lactose and protein concentrations and increased fat concentration. These effects, however, can vary widely depending on the type of restriction, its duration and intensity, or the stage of lactation in which it takes place. Finally, to avoid yield loss and metabolic disorders, it is important to identify reliable biomarkers to monitor energy balance.  相似文献   

番鸭产蛋各期肝脂肪酸组成及AMPK信号通路基因表达研究     

朱文俊  陈兴勇  刘乐  刘政权  赵羽彤  耿照玉 《畜牧兽医学报》2021,52(9):2429-2438

旨在探究产蛋各期番鸭肝腺苷酸活化蛋白激酶（AMPK）信号通路基因表达和肝脂肪酸组成,为番鸭肝脂质代谢应答产蛋提供机理研究。本研究选取开产前22周龄、产蛋初期30周龄、产蛋中期40周龄和产蛋末期60周龄母番鸭各15羽,全自动生化仪测定血脂水平,苏木精-伊红（HE）染色和油红O染色观察肝组织学结构,实时荧光定量PCR （qRT-PCR）检测肝AMPK通路基因表达,气相色谱-质谱联用法（GC-MS）检测产蛋各期肝脂肪酸组成。结果表明,总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、甘油三酯（TG）和极低密度脂蛋白胆固醇（VLDL-C）水平在40周龄显著高于22、30和60周龄（P<0.05）;高密度脂蛋白胆固醇（HDL-C）水平在30和40周龄显著低于22和60周龄（P<0.05）。肝HE和油红O染色切片显示,肝在22周龄呈实质状,至产蛋40和60周龄,肝脂滴沉积明显（P<0.05）。AMPKα1在22、30、40和60周龄呈本底低水平表达,并显著低于产蛋各期肉碱脂酰转移酶1（CPT1）和脂肪酸合成酶（FAS）的表达量（P<0.05）,FAS在22、40和60周龄均呈高水平表达（P<0.05）;羟甲基戊二酰辅酶A还原酶（HMGR）、肝细胞核因子4α（HNF4α）、乙酰辅酶A羧化酶1（ACC1）和固醇调控元件结合蛋白-1（SREBP1c）在40周龄表达量显著高于22和30周龄（P<0.05）。肝中饱和脂肪酸（SFA）、单不饱和脂肪酸（MUFA）和多不饱和脂肪酸（PUFA）主要由C16∶0、C18∶0、C18∶1和C20∶2 n-6构成,分别占总脂肪酸含量的32%、16%、30%和9%。C14∶0和C16∶0含量在40周龄显著高于22周龄（P<0.05）;C24∶0、C20∶2 n-6和PUFA含量在60周龄显著高于22和40周龄（P<0.05）。综上,番鸭产蛋期肝通过上调FAS等脂质合成基因表达,合成大量长链脂肪酸,沉积于肝,并增加血脂水平。  相似文献   

Differentially expressed genes identified through RNA‐seq with extreme values of principal components for beef fatty acid in Nelore cattle     

Bianca Ferreira Olivieri  Camila Urbano Braz  Fernando Brito Lopes  Elisa Peripolli  Rafael Medeiros de Oliveira Silva  Rosana Ruegger Pereira da Silva Corte  Lucia Galvo de Albuquerque  Anglica Simone Cravo Pereira  Nedenia Bonvino Stafuzza  Fernando Baldi 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2021,138(1):80-90

The aim of this study was to identify differentially expressed genes (DEG) in the Longissimus thoracis muscle of Nelore cattle related to fatty acid (FA) profile through RNA sequencing and principal component analysis (PCA). Two groups of 10 animals each were selected containing PC1 and PC2 extreme DEG values (HIGH × LOW) for each FA group. The intramuscular fat (IMF) was compared between cluster groups by ANOVA, and only the sum of monounsaturated FA (MUFA) and ω3 showed significant differences (p < .05). Interestingly, the highest percentage (95%) of phenotypic variation explained by the sum of the first two PC was observed for ω3, which also displayed the lowest number of DEG (n = 1). The lowest percentage (59%) was observed for MUFA, which also revealed the largest number of DEG (n = 66). Since only MUFA and ω3 exhibited significant differences between cluster groups, we can conclude that the differences observed for the remaining groups are not due to the percentage of IMF. Several genes that have been previously associated with meat quality and FA traits were identified as DEG in this study. The functional analysis revealed one KEGG pathway and eight GO terms as significant (p < .05), in which we highlighted the purine metabolism, glycolytic process, adenosine triphosphate binding and bone development. These results strongly contribute to the knowledge of the biological mechanisms involved in meat FA profile of Nelore cattle.  相似文献   

Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches     

Ana Carina Heil da Silva Matos  Angélica Consalter  Bárbara Paula dos Santos Batista  Ana Beatriz Monteiro Fonseca  Ana Maria Reis Ferreira  Juliana da Silva Leite 《Reproduction in domestic animals》2021,56(4):667-672

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest^® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.  相似文献   

Production,chemical composition,and fatty acid profile of milk from buffaloes fed with cupuaçu (Theobroma grandiflorum) cake and murumuru (Astrocaryum murumuru) cake in the Eastern Amazon     

André Guimarães Maciel e Silva  Suely Cristina Gomes de Lima  Pedro Danilo de Oliveira  Michel dos Santos Moraes  Célia Maria Costa Guimarães  Jamile Andréa Rodrigues da Silva  Alexandre Rossetto Garcia  Benjamim de Souza Nahúm  Lilaine de Sousa Neres  Gerlane Nunes Noronha  José de Brito Lourenço Júnior 《Animal Science Journal》2021,92(1):e13576

The objective was to evaluate the effect of concentrate supplementation using by-products of the Amazonian industry on milk production, milk composition, and milk fatty acid profile of dairy buffaloes. Twelve lactating buffaloes (544.5 ± 35.6 kg, 6.4 ± 2.2 years old, 59 ± 6 days in milk) were allotted in a pasture of Mombaça grass and managed under rotational grazing (4 days occupancy/28 days rest). A 3 × 3 Latin square was adopted, and each animal alternately received three supplementary treatments based on corn bran + soybean meal or cupuaçu cake or murumuru cake for 21 days per treatment. Murumuru cake increased the levels of lauric acid and myristic acid in the milk (p < 0.05). Murumuru cake reduced the unsaturated fatty acid contents in the milk compared with animals fed control diet or cupuaçu cake (24.27% vs. 25.24% vs. 25.08%). The n-6/n-3 ratio was 2.6, 1.97, and 2.0 in the control, cupuaçu, and murumuru groups, respectively. Based on this parameter, cakes made from cupuaçu as well as murumuru could be considered to be adequate for inclusion in dairy water buffalo feed. However, the murumuru cake addition requires some caution because its use induces the secretion of higher levels of lauric and myristic fatty acids that are related to human cardiovascular disease.  相似文献   

感染16型蓝舌病病毒后绵羊部分细胞因子消长特点研究     

苗海生  寇美玲  谢佳芮  李乐  李华春 《中国畜牧兽医》2021,48(1):296-302

本试验旨在探明绵羊感染16型蓝舌病病毒（Bluetongue virus type 16,BTV16）后细胞因子IFN-γ、IL-2、IL-4和IL-10的消长特点。用实时荧光定量PCR检测方法对感染BTV16后3只绵羊上述4种因子mRNA进行检测,同时设立阴性对照绵羊,并以0 d mRNA为基准,计算mRNA的相对表达量,同时检测病毒抗体效价、测量绵羊体温。结果显示,接种BTV16的3只绵羊均不同程度产生抗体和体温症状,4种细胞因子的mRNA在接种病毒2~4 d内均出现显著上升,其中IFN-γ峰值在2.58~27.84倍之间,IL-2峰值在5.24~17.19倍之间,IL-4峰值在2.16~3.43倍之间,IL-10峰值在15.78~48.77倍之间,个体上升幅度存在显著差异,4种细胞因子均在高水平持续6 d左右后逐渐下降。对照绵羊上述参数在正常范围内波动。本研究阐明了接种BTV16后绵羊细胞因子IFN-γ、IL-2、IL-4、IL-10在转录水平上的消长特点,为进一步深入开展BTV感染特征、宿主机体免疫机制研究提供参考。  相似文献   

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究     

常雯茹  段利芳  杨乐  马英先  王棋  翟云云  周鲁豫  张爽  明胜利  杨国宇  王江  褚贝贝 《中国畜牧兽医》2021,(1)

试验旨在研究伪狂犬病病毒(PRV)在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞(3D4/21)p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。  相似文献   

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究     

常雯茹  段利芳  杨乐  马英先  王棋  翟云云  周鲁豫  张爽  明胜利  杨国宇  王江  褚贝贝 《中国畜牧兽医》2021,48(1):83-92

试验旨在研究伪狂犬病病毒（PRV）在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞（3D4/21）p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。  相似文献   

外源肌苷酸对种鸡产蛋性能、蛋品质及种蛋孵化性能的影响     

牛自康  吴乐  张鑫蕊  叶成智  宦海琳  张峰  魏明星  闫俊书 《中国畜牧兽医》2021,(1)

本试验通过在种鸡的基础饲粮中加入肌苷酸(IMP)来研究其对种鸡产蛋性能、蛋品质和种蛋孵化性能的影响。选择遗传背景相同、体重相近,产蛋率达到5%的20周龄健康AA种鸡864羽,随机分为2组(对照组饲喂基础饲粮;试验组饲喂基础饲粮+0.5%肌苷酸),每组6个重复,每个重复72羽,试验期30 d。测定试验期间所有种蛋的平均蛋重、产蛋率和合格蛋率;每组随机选取部分种蛋进行蛋品质测定;于孵化开始第17天,通过照蛋处理统计种蛋受精率,第21天出雏时,计算受精蛋孵化率、入孵蛋孵化率和健雏率。试验结果表明:与对照组相比,试验组种鸡的平均蛋重、产蛋率和合格蛋率均有升高的趋势,但差异不显著(P>0.05);蛋形指数显著升高(P<0.05),蛋壳强度、蛋壳厚度和蛋黄颜色显著降低(P<0.05);受精率、受精蛋孵化率和入孵蛋孵化率显著升高(P<0.05),健雏率和雏鸡体重有升高的趋势,但差异不显著(P>0.05)。综上,0.5%外源肌苷酸对种鸡的产蛋性能和种蛋孵化性能均有促进作用。  相似文献