牛星状病毒EvaGreen实时荧光定量PCR检测方法的建立及应用

牛星状病毒(BAstV)是我国新发的犊牛腹泻病原,本试验的目的是建立检测BAstV的Real-time PCR方法。根据BAstV流行株的ORF1a基因序列设计引物,通过优化反应条件和体系,成功建立基于EvaGreen检测BAstV的Real-time PCR方法。结果表明,该检测方法的Ct值与标准品模板在1.36×10¹~1.36×10⁸拷贝/μL线性关系良好,相关系数R²=0.999,扩增效率为93.79%;该方法可特异性检出BAstV,对犊牛腹泻其他相关病原呈阴性;最低检测下限为13.6拷贝/μL;批间和批内的变异系数均小于2%,重复性好。对2017年9月至2019年5月采自河南省的221份犊牛腹泻样本进行检测,BAstV的检出率为18.1%(40/221),采样场阳性率为100.0%(14/14)。本试验所建方法灵敏度高、特异性强、稳定性好,为BAstV的检测和流行病学调查提供了有力手段。     

Random-effects linear model application to herd-level assessment of bovine hepatic trace mineral concentrations

To evaluate the utility of random-effects linear modeling for herd-level evaluation of trace mineral status, we performed a retrospective analysis of the results for trace mineral testing of bovine liver samples submitted to the Michigan State University Veterinary Diagnostic Laboratory between 2011 and 2017. Our aim was to examine random-effects models for their potential utility in improving interpretation with minimal sample numbers. The database consisted of 1,658 animals distributed among 121 herds. Minerals were assayed by inductively coupled plasma–mass spectroscopy, and included cobalt, copper, iron, molybdenum, manganese, selenium, and zinc. Intraclass correlation coefficients for each mineral were significantly different (p < 0.001) from zero and ranged from 0.38 for manganese to 0.82 for selenium, indicating that the strength of herd effects, which are presumably related to diet, vary greatly by mineral. Analysis of the distribution and standard errors of best linear unbiased predictor (BLUP) values suggested that testing 5–10 animals per herd could place herds within 10 percentile units across the population of herds with 70–95% confidence, the confidence level varying among minerals. Herd means were generally similar to BLUPs, suggesting that means could be reasonably compared to BLUPs with respect to the distributions reported here. However, caution in interpreting means relative to BLUPs should be exercised when animal numbers are small, the standard errors of the means are large, and/or the values are near the extremes of the distribution.     

口蹄疫疫苗对牛的应用效果调查

牛口蹄疫是传染性疾病,为研究青海省峰堆乡口蹄疫疫苗的免疫效果,于2018—2019年对该乡的7个村开展牛口蹄疫疫苗免疫效果调查,共调查牛634头,7个村总体的口蹄疫合格率为100.00%和97.56%,最低合格率为81.63%和88.63%。为更好地控制该类疾病发生流行,提升免疫效果,该文探讨口蹄疫疫苗免疫失败原因,并制定了相应的预防措施。     

蜂胶对细菌脂多糖刺激下奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接蛋白的影响

本试验旨在研究中国蜂胶乙醇提取物（ethanol extract of Chinese propolis，EECP）对细菌脂多糖（lipopolysaccharide，LPS）刺激下体外培养奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接渗透性的影响。EECP中总酚酸和总黄酮含量测定采用福林酚法和硝酸铝法，并建立LPS诱导奶牛乳腺上皮细胞（bovine mammary epithelial cells，MAC-T）炎症模型，采用CCK-8法测定EECP对MAC-T相对增殖率的影响，利用实时荧光定量PCR（RT-qPCR）评估EECP对LPS诱导的MAC-T细胞炎症相关因子（IL-6、IL-8、TNF-α和IL-1β）相对mRNA转录水平；以及对紧密连接蛋白（occludin、ZO-1）相对mRNA转录水平进行检测，并进一步利用免疫荧光技术对紧密连接膜蛋白进行定位，确定EECP对LPS诱导MAC-T细胞炎症紧密连接渗透性的影响。结果显示：EECP中总酚酸含量为106.35 mg没食子酸当量（GAE）·g^-1、总黄酮含量为320.85 mg芦丁当量（RE）·g^-1；CCK-8结果显示EECP的安全浓度为0~15 μg·mL^-1，并可有效提高LPS刺激下MAC-T的活力；LPS刺激显著增加了细胞炎症相关因子IL-6、IL-8、TNF-α和IL-1β mRNA的转录量（P<0.001）；但2.5~15.0 μg·mL^-1 EECP预处理显著降低了IL-6、IL-8、TNF-α和IL-1β mRNA的转录量；与此类似，LPS刺激显著抑制了紧密连接蛋白基因（occludin、ZO-1）mRNA的转录量（P<0.01），而EECP预处理后紧密连接蛋白基因（occludin和ZO-1）mRNA的转录量显著增加（P<0.05）；免疫荧光染色试验也证实EECP能通过上调紧密连接蛋白（occludin、ZO-1）的表达，缓解LPS诱导的乳腺上皮细胞屏障功能紊乱。该结果证实，EECP对细菌脂多糖诱导奶牛乳腺上皮细胞炎症具有良好的保护作用，这为利用中国蜂胶预防奶牛乳腺炎提供了试验基础。     

Cellular activity and development of enteric,hepatic and muscle tissues of juvenile pacu Piaractus mesopotamicus (Holmberg 1887) fed with lyophilized bovine colostrum

Bovine colostrum, an alternative food of animal origin provided as partial protein source, may constitute an innovative ingredient in fish feeding. Serum insulin‐like growth factor 1 (IGF‐1) and cellular activity of enteric, hepatic and muscle tissues of juvenile pacu Piaractus mesopotamicus fed with lyophilized bovine colostrum (LBC) were evaluated. Pacu (8.46 ± 0.74 g; 7.75 ± 0.27 cm, n = 18 in triplicate) were stocked into 18 tanks (300 L; 18 fish per tank, in triplicate) and fed with control diet containing 0% of LBC or experimental diet containing 10% and 20% of LBC inclusion for either 30 or 60 days of experiment. IGF‐1 serum concentration was not affected (P > 0.05) by the diet containing LBC and experimental period. The intestinal and hepatic cellular activity was not influenced by LBC (P > 0.05); however, intestine protein/RNA decreased at 60 days (P < 0.05). The muscle cellular activity was affected by LBC, highest in fish fed with diet containing 10% of LBC (P < 0.05). Between the periods DNA and protein/RNA increased at 60 days, while RNA, protein/DNA and RNA/DNA ratio decreased in this same period (P < 0.05). Increased DNA muscle content indicates fish growth by hyperplasia. The results of the present study is related with suitable development of enteric, hepatic and muscle tissues and proper condition of juvenile pacu fed with LBC considering the absence of differences in relation to juveniles that consumed diet without colostrum.     

牛出血性败血症诊断与防治

牛出血性败血症具有致病因素多、传染源多、危害大的特点,养殖户要针对该病的特点制定有效防控方案,并严格执行防控措施,定期对牛群给予免疫注射,并加强饲养管理,才能有效降低发病几率,提升经济效益。该文主要论述牛出血性败血症的诊断与防治。     

牛胃肠炎发生原因及防治

胃肠炎是家畜生长中的常见病和高发病,牛具有很高的发病率。造成胃肠炎发生的原因是饲养管理不到位、饲料营养价值较差,饲料不足,牛出现营养不良,身体抵抗能力下降,使消化机能变差,进入消化道中的饲料不能很好的吸收,肠鸣音大,饲料随着肠道的蠕动排出体外。临床上主要表现为腹泻、身体脱水、生长发育不良。由于牛胃肠炎的发生机理还不明确,病因比较复杂,在疾病诊治期间应尝试明确实际的发病愿意,对动物进行对症防治,确保牛健康生长。该文主要论述牛胃肠炎的发生原因和防治措施。     

Antioxidant,anti-amylase and anti-glycation potential of brans of some Sri Lankan traditional and improved rice (Oryza sativa L.) varieties

Brans of 23 traditional and 12 improved (both red and white) rice varieties in Sri Lanka were screened for anti-amylase and anti-glycation activities in vitro. Varieties which showed the highest inhibitory activities at screening were further investigated for anti-glucosidase and glycation reversing as anti-diabetic properties. The same varieties were studied for selected antioxidant properties. Significantly high anti-amylase and anti-glycation activities were observed for bran extracts of red varieties compared to white varieties at screening. Traditional red rice varieties, Masuran, Sudu Heeneti, Dik Wee and Goda Heeneti, exhibited significant and dose dependent anti-amylase, anti-glycation and glycation reversing activities. These varieties also showed marked antioxidant properties. It is concluded that brans of Sri Lankan traditional red rice varieties Masuran, Sudu Heeneti, Dik Wee and Goda Heeneti may be potential food supplements for diabetes.     

Genetic characterization of bovine viral diarrhea virus strains in Beijing,China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle

To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5''-untranslated region (5''-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV.