猪急性腹泻综合征冠状病毒SYBR Green荧光定量PCR检测方法的建立及应用     

张记宇  韩郁茹  时洪艳  陈建飞  张鑫  刘建波  张燎原  冯书风  冯廷帅  季朝阳  石达  冯力 《畜牧兽医学报》2021,52(10):2887-2894

为建立快速、灵敏且特异的检测猪急性腹泻综合征冠状病毒（swine acute diarrhea syndrome coronavirus,SADS-CoV）检测方法,本试验扩增SADS-CoV N基因保守区域将其克隆至pMD18-T载体。所构建的重组质粒pMD18-T-SADS-qN作为阳性质粒标准品,以其为模板建立一种SYBR Green荧光定量PCR检测方法。结果显示,所建立方法在3.31×10¹~3.31×10⁷拷贝·μL^-1模板量时,呈良好的线性关系,相关系数（R²）为0.997,斜率为-3.318。该方法特异性检测SADS-CoV;而猪流行性腹泻病毒（PEDV）、猪传染性胃肠炎病毒（TGEV）、猪德尔塔冠状病毒（PDCoV）和猪繁殖与呼吸综合征病毒（PRRSV）检测结果均为阴性。所构建的标准品检测灵敏度下限可以达到3.31×10¹拷贝·μL^-1,组内和组间变异系数均小于1%,表明其具有良好的灵敏性和重复性。用该方法检测SADS-CoV感染IPI-2I和IPEC-J2细胞后不同时间点和不同接毒剂量的复制情况,结果显示,SADS-CoV感染细胞后2 h病毒含量较低,在12~36 h病毒含量迅速增长,36 h后增长速度减缓且病毒含量维持在较高水平。分别用0、0.1、1 MOI SADS-CoV感染细胞结果显示病毒的mRAN转录水平呈现剂量依赖性增加,当MOI为1时,IPI-2I和IPEC-J2细胞病毒含量分别为10^6.7、10^5.3拷贝·mL^-1。进一步利用所建立的方法对经口服攻毒SADS-CoV仔猪的临床样本进行检测,结果发现病毒在空肠回肠含量较高,表明病毒主要定殖于空肠和回肠。综上表明,本研究建立SYBR Green荧光定量PCR检测方法能灵敏特异地检测SADS-CoV,为SADS-CoV的诊断和病毒相关基础研究提供可靠的检测手段。  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   

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination     

Aline Fernanda Lopes Paschoal  Ana Paula Gonçalves Mellagi  Cristina Vicenti Ferrari  Karine Ludwig Takeuti  Gabriela da Silva Oliveira  Mari Lourdes Bernardi  Rafael da Rosa Ulguim  Fernando Pandolfo Bortolozzo 《Reproduction in domestic animals》2021,56(6):897-904

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.  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   

Free-range system and supplementation of 25-hydroxicholecalciferol increases the performance and serum vitamin levels in mixed-parity sows     

Josiane C. Panisson  Nathália C. Oliveira  Cristina M. Sá-Fortes  Adsos A. Passos  Claudia C. da Silva  Wagner A. G. Araújo  Idael M. G. Lopes  Gleison M. S. Costa  Iara Q. Ataíde  Bruno A. N. Silva 《Animal Science Journal》2021,92(1):e13592

Improvements in sow productivity have raised questions regarding dietary vitamin D recommendations. The present study aimed to evaluate the effects of the housing system with access to sunlight exposure and supplementation of 25-hydroxicholecalciferol on performance and serum levels of 25(OH)D₃ in sows during gestation and lactation. Sows were distributed in an experimental design with two housing systems: gestation crates or gestation free-range system with external area for sunlight exposure; and two diets: 0 or 50 μg of 25-hydroxicholecalciferol kg⁻¹. The use of 25-hydroxicholecalciferol tended (P = 0.052) to improve total born and influenced (P = 0.046) on number of born alive. Litter weight at birth was also increased (P = 0.01) by 25-hydroxicholecalciferol supplementation; 25-hydroxicholecalciferol supplementation and housing system (free-range with sunlight exposure) tended to increase weaning weight (P = 0.07) and litter daily gain (P = 0.051) during lactation. Exposure to sunlight and 25-hydroxicholecalciferol supplementation increased 25(OH)D₃ serum levels when compared with control treatment during gestation (136.95 vs. 113.92 ng mL⁻¹; P = 0.035) and lactation (120.29 vs. 88.93 ng mL⁻¹; P = 0.026). In conclusion, the association of 25-hydroxicholecalciferol supplementation with exposure to sunlight during gestation improved significantly 25(OH)D₃ serum levels and consequently performance traits in gestation and lactation.  相似文献   

冬小麦种质“矮孟牛”中新型小麦-黑麦复杂易位的遗传传递分析   总被引：8，自引：3，他引：8  

亓增军  刘大钧  陈佩度  王苏玲  李斯深  李晴祺 《作物学报》2001,27(5):582-589

本文利用矮孟牛中一种新型小麦-黑麦复杂易位IRS·7DS,1BL·7DL与1BL·1RS易位所构建的一套重组自交系(包括109个F6株系)和8个矮孟牛衍生品种进行了分析,结果表明在109个F6株系中,纯合复杂易位占69个,纯合1BL·1RS占36个,二者比例约2∶1,杂合类型占4个,出现频率4/109;对矮孟牛8个衍生品种的分析表明,6个品种包含来自矮孟牛的  相似文献   

Effect of the Nixtamalization Process on the Dietary Fiber Content,Starch Digestibility,and Antioxidant Capacity of Blue Maize Tortilla     

Luis A. Bello‐Prez  Pamela C. Flores‐Silva  Gustavo A. Camelo‐Mndez  Octavio Paredes‐Lpez  Juan de Dios Figueroa‐Crdenas 《Cereal Chemistry》2015,92(3):265-270

Nixtamalization is an ancient process developed by the Mesoamerican cultures. Initially, volcanic ashes were used and then calcium hydroxide in commercial production, and more recently nixtamalization with calcium salts (NCS) has been proposed. The aim of this study was to evaluate the effect of NCS on carbohydrate digestibility and antioxidant capacity in the elaboration of blue maize tortillas. NCS in blue tortillas showed a high amount of total dietary fiber (14.27 g/100 g), the main fraction being insoluble dietary fiber. The contents of resistant starch and slowly digestible starch did not change with the nixtamalization process. The predicted glycemic index value was lower in blue tortillas with the NCS process (58) than with the traditional nixtamalization process (71). In general, NCS in blue tortillas presented a higher antioxidant capacity than traditional tortillas (ferric reducing antioxidant power method), indicating that phenolics present in blue maize maintain their activity after cooking. It can be concluded that the nutraceutical features (high dietary fiber content and antioxidant capacity) of blue maize tortillas are enhanced when they are elaborated with the NCS process.  相似文献   

Tear ferning test in horses and its correlation with ocular surface evaluation          下载免费PDF全文

Laura R. Silva  Anderson F. Gouveia  Cleyber J. T. de Fátima  Letícia B. Oliveira  Janildo L. Reis Jr.  Renato F. Ferreira  Concepta M. Pimentel  Paula D. Galera 《Veterinary ophthalmology》2016,19(2):117-123

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Intake,performance, and carcass characteristics of lambs fed spineless cactus replacing wheat bran     

Sabrina Carla Rodrigues Felix  Ricardo Alexandre Silva Pessoa  Marcelo de Andrade Ferreira  Luciana Felizardo Pereira Soares  Janaina de Lima Silva  Karen Santos Felix de Abreu  Ana Caroline Cerqueira de Melo 《Tropical animal health and production》2016,48(2):465-468

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