广东省牛流行热病毒分离株JM 2020的全基因组测序及进化分析

【目的】分析广东省牛流行热病毒(Bovine ephemeral fever virus,BEFV)分离株JM 2020与其他地区毒株的进化关系,阐明遗传进化与全基因组的特征,为中国乃至世界对牛流行热疾病的流行情况以及预防该疾病提供有用信息。【方法】根据从GenBank下载的BEFV毒株的全基因组序列信息,设计引物PCR扩增糖蛋白(Glycoprotein,G)基因;另设计10对引物用于扩增全基因组,送测序得到10个片段的基因序列,运用DNAStar软件中的EditSeq手动编辑和拼接获得全基因组序列。运用MEGA 6.0分别构建G基因和全基因组进化树进行进化分析。【结果】JM 2020毒株的G基因和全基因组与泰国毒株的核苷酸序列相似性均最高,分别为94.9%～99.3%和99.0%,进化分析表明JM 2020与2013—2017年泰国毒株处于同一小分支,而与JB76H、JT02L等国内毒株有一定进化距离。全基因组测序结果表明,JM 2020全基因组长度为14 902个核苷酸(nt),包括50 nt的前导序列,1 296 nt的核蛋白(Nucleoprotein,N)基因,837 nt...     

Immunohistochemical expression of interferon-γ in different types of granulomatous lesions associated with bovine paratuberculosis

Animals infected with Mycobacterium avium subsp. paratuberculosis (Map) show a variety of lesions, from focal forms, seen in subclinical stages to diffuse lesions in clinical cases. The purpose of this study was to evaluate the local expression of IFN-γ by immunohistochemistry in relation with the type of lesion in naturally Map-infected cows. The number of immunolabelled cells, −the majority morphologically consistent with lymphocytes-, was higher in focal and diffuse paucibacillary forms than in diffuse multibacillary lesions, where they appeared closely related to epithelioid cells. Diffuse multibacillary lesions had the lowest numbers, but higher than controls, and positive cells were intermingled among the macrophages. The peripheral IFN-γ production was higher in all Map infected cows and a positive correlation was found with the number of immunolabelled cells in the intestine. The findings of this study show that IFN-γ would play a role in the development of the different types of lesions in paratuberculosis, and also points out the importance of adequate sampling of lymphoid tissue containing samples when studying the local immune response in which IFN-γ expression may be involved, especially in cases where focal lesions are present.     

Tricuspid valve dysplasia and a patent foramen ovale resulting in severe tricuspid regurgitation and right-heart dilation in a Red Angus calf

A two-month-old Red Angus heifer calf presented to the University of Wisconsin Veterinary Care for evaluation of suspected severe bronchopneumonia. Pertinent physical exam findings included tachycardia, tachypnea, dyspnea with a significant abdominal component, and cyanotic mucous membranes. On thoracic auscultation, wheezes were present bilaterally, as well as a grade 2/6 right apical systolic murmur. Thoracic radiographs revealed cardiomegaly, most severely affecting the right side. Echocardiography showed tricuspid valve dysplasia, resulting in severe tricuspid regurgitation and right-heart dilation, as well as a patent foramen ovale. A postmortem examination confirmed the presence of the aforementioned cardiac abnormalities and revealed only mild pulmonary changes. This case report is the first to describe tricuspid dysplasia in the absence of multiple, complex congenital cardiac abnormalities in a calf, and it highlights the value of echocardiography for an antemortem diagnosis.     

不同导血水平对夏南牛生长发育的影响

本研究把夏南牛各代次、各年龄段牛与南阳牛之间体重做了比较分析;同时与原保留对比试验组的含夏洛来牛血50%和25%的两组横交牛进行比较,结果表明:新品种牛公牛的0、6、12、18月龄四个年龄段与南阳牛相比,体重差异均为极显著(P<0.01).母牛在O、6、12、18、24、30、36月龄各年龄段与南阳牛相比,体重差异均为极显著(P0.05).     

Spoligotype diversity of Mycobacterium bovis and Mycobacterium caprae animal isolates

The genetic diversity of 283 Mycobacterium bovis (M. bovis) and 10 Mycobacterium caprae (M. caprae) strains, isolated between 2002 and 2007 from cattle, goat, red deer and wild boar from six different geographical regions of Portugal was investigated by spoligotyping. The technique showed a good discriminatory power (Hunter-Gaston Index, h=0.9) for the strains, revealing 29 different patterns. One pattern (SB0121) was clearly predominant, accounting for 26.3% of the isolates; ten patterns, representing 20.7% of the isolates, had never been reported previously. Multiple spoligotypes were detected in thirteen cattle and one goat herd, most of which were found in beef cattle and extensive management regions, suggesting different infection sources. With the exception of two spoligotypes, those in wildlife species were also found in domestic species.     

Localization of leptin and leptin receptor in the bovine adenohypophysis

The present study was carried out to detail the cellular localization of leptin (Lep) and the leptin receptor (LepR) in the bovine adenohypophysis. Lep immunoreactivity (Lep-ir) was found in about 30% of adenohypophysial cells in the gland. Immunochemistry of Lep and specific hormones using serial sections revealed that Lep-ir was present in 60.4% of somatotrophs, 15.9% of gonadotrophs, 6.5% of mammotrophs, 6.5% of thyrotrophs and 2.4% of corticotrophs. Both the common short isoform (OBRa) and the long isoform (OBRb) of LepR mRNA were expressed in the bovine adenohypophysis. LepR immunoreactivity (LepR-ir) was found in only 2.8% of the adenohypophysial cells and over 50% of LepR-ir cells were gonadotrophs, in which most of the cells were distributed in the zona tuberalis. The findings on Lep and LepR in the adenohypophysial cells indicate that Lep may regulate gonadotroph function through autocrine/paracrine pathway in the bovine adenohypophysis.     

Renal amyloidosis in cattle with inflammatory diseases

BACKGROUND: The association of inflammatory diseases such as traumatic reticuloperitonitis (TRP), mastitis, metritis, and pododermatitis with renal amyloidosis in cattle is poorly described. HYPOTHESIS: Serum amyloid A (SAA) levels are elevated during inflammatory diseases, and renal amyloidosis is formed as a complication. ANIMALS: This study was conducted with 82 crossbred cattle with mastitis (n = 18 cows), metritis (n = 11 cows), TRP (n = 30 cows), and pododermatitis (n = 23 : 15 cows and 8 beef cattle). Ten clinically healthy cows served as controls. Methods: Hematological, urinary, and blood parameters, including SAA, were measured by an automated procedure provided with trade kits. Determination of amyloidal structures was made by histopathological examination of renal biopsy specimens. RESULTS: At the end of this trial, amyloidosis was detected in 5 cows displaying typical nephrotic syndrome, with hypoproteinemia and proteinuria in combination with polyuria and weight loss. Furthermore, it was observed that cows with renal amyloidosis had significantly higher (P < .01) total leukocyte counts, serum and urine enzyme activities, and urea and creatinine concentrations, with lower serum total protein concentrations, when compared with animals without renal amyloidosis. CONCLUSIONS AND CLINICAL IMPORTANCE: The incidence of AA amyloidosis in cattle in this study suggests that cattle with mastitis, metritis, and pododermatitis have a high prevalence of systemic amyloidosis in response to inflammation.     

牛杀菌／通透性增加蛋白氮端基因的克隆和序列分析

本试验应用RT-PCR技术,参照Genbank报道的序列,从荷斯坦牛中性粒细胞mRNA中扩增出杀菌／通透性增加蛋白（BPI）氮端基因（713bp）,并与pGEM-T-easy载体连接,构建基因重组体pGEM-T-easy-BPI,进行序列测定。结果表明获得长度为713bp的BPI氮端基因。序列分析证实该片段与安哥斯牛BPI氮端基因相比有1个点突变,为同义突变。该基因的克隆为进一步表达该基因奠定了基础。     

牛卵母细胞体外成熟技术研究

从两方面探讨了牛卵母细胞的体外成熟培养效果,即对不同级别和不同卵巢卵泡直径的卵丘卵母细胞复合体（COCs）进行成熟培养,测定其成熟率。COCs级别不同,培养成熟率之间存在很大差异,A级COCs成熟率为63．81％,B级为47．81％,显著高于C级和D级（P〈0．01）。卵母细胞的成熟率与采集COCs时的卵泡直径关系密切。中等卵泡的卵母细胞的成熟率为61．82％,显著高于小卵泡和大卵泡（P〈0．01）。试验结果表明：A级和B级COCs是卵母细胞体外培养的主要资源;在卵母细胞体外培养过程中,宜选择卵泡发育水平基本正常的中等卵泡采集COCs,而不宜选择发育过大或过小的卵泡。