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奶牛乳腺炎5种病原菌多重PCR快速检测方法的建立与评价 被引量:4
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作者 申纪饶 王丹 +7 位作者 李新圃 杨峰 武小虎 丁学智 王胜义 严作廷 王旭荣 李宏胜 《中国预防兽医学报》 CAS CSCD 北大核心 2021年第5期507-511,533,共6页
为建立一种能同时快速检测多种奶牛乳腺炎致病菌的多重PCR检测方法,本研究以化脓隐秘杆菌ISR基因、无乳链球菌cfb基因、大肠杆菌phoA基因、副乳房链球菌23S rRNA基因和金黄色葡萄球菌nuc基因为靶基因,设计并合成了5对特异性引物,通过方... 为建立一种能同时快速检测多种奶牛乳腺炎致病菌的多重PCR检测方法,本研究以化脓隐秘杆菌ISR基因、无乳链球菌cfb基因、大肠杆菌phoA基因、副乳房链球菌23S rRNA基因和金黄色葡萄球菌nuc基因为靶基因,设计并合成了5对特异性引物,通过方阵法对多重PCR反应体系及反应条件优化,建立了一种能快速检测5种病原菌的多重PCR方法。对各菌种随机组合后利用该多重PCR方法检测,结果显示,该方法能同时快速检测奶牛乳腺炎乳样中5种主要病原菌,而其他病原菌则呈阴性结果,具有较强特异性。分别将化脓隐秘杆菌、无乳链球菌、大肠杆菌、副乳房链球菌、金黄色葡萄球菌菌液10倍倍比稀释后,利用建立的多重PCR方法进行检测,确定该方法敏感性,结果显示,该方法对5种病原菌最低检测浓度分别为:金黄色葡萄球菌6.25×10^(4)cfu/mL,大肠杆菌2.95×10^(6)cfu/mL,化脓隐秘杆菌2.95×10^(5)cfu/mL,副乳房链球菌4.3×10^(5)cfu/mL,无乳链球菌3.15×10^(5)cfu/mL。对临床送检及人工模拟的乳样检测结果显示该方法具有较好符合率及灵敏度。本研究首次建立了对包括副乳房链球菌在内的奶牛乳腺炎致病菌多重PCR检测方法,为快速检测奶牛乳腺炎病原菌提供了可行的技术手段。 展开更多
关键词 奶牛乳腺炎 病原菌 引物 反应条件 多重PCR
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Penicillin-resistant characterization of Staphylococcus aureus isolated from bovine mastitis in Gansu,China 被引量:3
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作者 yanG Feng LIU Long-hai +7 位作者 WANG Ling WANG Xu-rong LI Xin-pu LUO Jin-yin ZHANG Zhe ZHANGShi-dong yan zuo-ting LI Hong-sheng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第8期1874-1878,共5页
Bovine mastitis caused by Staphylococcus aureus is difficult to treat because of increasing resistance against antibiotics, especially penicillin. β-Lactamase and biofilm are responsible for penicillin resistance of ... Bovine mastitis caused by Staphylococcus aureus is difficult to treat because of increasing resistance against antibiotics, especially penicillin. β-Lactamase and biofilm are responsible for penicillin resistance of S. aureus. The aim of this study was to investigate the β-lactamase activity and biofilm formation capacity of 37 penicillin-resistant S. aureus strains (35 were blaZ positive and 2 were blaZ negative) from bovine mastitis in Gansu Province, China, as well as to measure the intercellular adhesion genes icaA and icaD of these strains. β-Lactamase Test Kit was used to determine the β-lactamase activity, biofilm formation was tested by semi-quantitative adherence assay method. Moreover, the presence of icaA and icaD were measured by PCR. A total of 32 penicillin-resistant S. aureus strains, including the two blaZ-negative strains, were identified as β-lactamase producers. All tested S. aureus isolates produced biofilm in the microtiter plate assay. Meanwhile, all these strains were PCR-positive for the ica locus, icaA and icaD. The study indicated high prevalence of β-lactamase activity, biofilm-forming capacity, and the ica genes among the penicillin-resistant S. aureus isolates, and implied that S. aureus resistant to penicillin was attributed to multiple mechanisms. 展开更多
关键词 Staphylococcus aureus penicillin-resistant Β-LACTAMASE BIOFILM ICA
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Prevalence and characteristics of extended spectrum β-lactamase-producing Escherichia coli from bovine mastitis cases in China 被引量:2
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作者 yanG Feng ZHANG Shi-dong +4 位作者 SHANG Xiao-fei WANG Xu-rong WANG Ling yan zuo-ting LI Hong-sheng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第6期1246-1251,共6页
The aim of the study was to investigate the prevalence and characterization of extended-spectrum β-lactamase (ESBL)- producing Escherichia coli isolated from bovine mastitis cases in China. ChromID ESBL agar was us... The aim of the study was to investigate the prevalence and characterization of extended-spectrum β-lactamase (ESBL)- producing Escherichia coli isolated from bovine mastitis cases in China. ChromID ESBL agar was used to confirm ESBL-producing E. coli. PCR and DNA sequencing were employed to characterize the genotype of ESBL-producers. Antimicrobial susceptibility was measured by disc diffusion. Overall, 73 of 318 E. coli isolates (22.96%) were identified as ESBL-producers. Of these ESBL-producing E. coli, the prevalence of blaCTX-M and blaTEM-1 was 97.26 and 71.23%, respectively. The predominant CTX-M-type ESBL was CTX-M-15 (65.75%), followed by CTX-M-14 (10.96%), CTX-M-55 (9.59%), CTX-M-64 (5.48%), CTX-M-65 (4.11%) and CTX-M-3 (1.37%). This study is the first report of CTX-M-64 and CTX-M-65 in E. coli isolated from bovine mastitis. Furthermore, 72 ESBL-producing E. coli isolates (98.63%) were found to be multidrug-resistance. This study noted high prevalence and rates of antimicrobial resistance of ESBL-producing E. coli isolates from bovine mastitis cases in China. 展开更多
关键词 extended spectrum β-lactamase Escherichia coli MULTIDRUG-RESISTANCE bovine mastitis
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中兽药参黄颗粒的制备工艺优化研究
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作者 柯群华 吕亚楠 +3 位作者 彭晶 吴雪琴 严作廷 王胜义 《中国兽药杂志》 2022年第5期53-62,共10页
为了优化中兽药参黄颗粒的制备工艺,获得最佳制备工艺参数,采用正交试验设计优化水提,浓缩,并采用湿法制粒,通过单因素试验和正交试验优选药物与辅料的比例、乙醇浓度和烘干温度参数,以颗粒的成型性、堆密度、休止角、溶化性以及吸湿性... 为了优化中兽药参黄颗粒的制备工艺,获得最佳制备工艺参数,采用正交试验设计优化水提,浓缩,并采用湿法制粒,通过单因素试验和正交试验优选药物与辅料的比例、乙醇浓度和烘干温度参数,以颗粒的成型性、堆密度、休止角、溶化性以及吸湿性来评价颗粒的最佳成型工艺。最佳水提工艺为加9倍水,回流提取3次,每次提取时间1 h;药物与辅料比例为(3∶5)药物浸膏粉∶可溶性淀粉∶糊精∶蔗糖为(3∶2∶2∶1),乙醇浓度为80%,烘干温度为60℃,制备的颗粒剂综合评分最高。中兽药参黄颗粒的制备工艺优化研究为其临床试验的扩大生产提供了科学参考依据。 展开更多
关键词 参黄颗粒 正交实验 综合评分 工艺优化
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Effect of Chanfukang(Chinese Medicine)on Endothelin and Nitric Oxide in Postpartum Cows with Qi Deficiency and Blood Stasis
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作者 yan zuo-ting WANG Dong-sheng +4 位作者 yanG Guo- lin XIE Jia-sheng LI Shi-hong ZHANG Shi-dong GONG Zhong-fu 《Animal Husbandry and Feed Science》 CAS 2010年第3期39-42,共4页
[ Objective] To observe effect of Chanfukang on vascular endothelial cells of postpartum cows with Oi deficiency and Blood stasis, and to explore the treatment mechanism of Chanfukang. [Method] A total of 58 cows were... [ Objective] To observe effect of Chanfukang on vascular endothelial cells of postpartum cows with Oi deficiency and Blood stasis, and to explore the treatment mechanism of Chanfukang. [Method] A total of 58 cows were assigned to four groups. The cows in group 1 and group 2 suffered from Qi deficiency and Blood stasis. The cows in group 1 were gavaged with Chanfukang (0.5 g/kg-BW, 5 d) at 12 h post partum, but the cows in group 2 were not treated. The cows in group 3 were healthy and gavaged with Chanfukang (0.4 g/kg.BW, 5 d) at 12 h post partum. Cows in group 4 were healthy control cows. The plasma endothelin (ET) concentration and serum nitric oxide (NO) concentration were determined before and after parturition, respectively. [ Result ] The plasma ET concentration and serum NO concentration were significantly higher in the cows with Qi deficiency and Blood stasis than in the healthy control cows (P 〈 0.05). The NO concentration decreased on Day 10 post partum in the cows with Qi deficiency and Blood stasis, but it was still higher than that in the healthy control cows. After taking Chanfukang for 7 d, the ET concentration and NO concentration decreased, and no significant difference was found between the cows with Qi deficiency and Blood stasis treated with Chanfukang and the healthy control cows. [ Conclusion] Chanfukang may relieve Qi deficiency and Blood stasis of postpartum cows through decreasing plasma ET concentration and serum NO concentration. 展开更多
关键词 Chanfukang Qi deficiency and Blood stasis Dairy cows ENDOTHELIN Nitric oxide
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