1株鸡毒支原体的分离鉴定及体外敏感中药的筛选

doi:10.16431/j.cnki.1671-7236.2021.03.031

Abstract

Abstract: The study was aimed to isolate,cultivate,identify and purify the prevalent strains of Mycoplasma gallisepticum in Tai'an,Shandong,and to screen sensitive traditional Chinese medicines against the isolates in order to provide materials for further research.The methods of liquid culture and solid culture were used to isolate,culture and purify the Mycoplasma gallisepticum strain from the tissue samples of diseased chickens,the isolate was preliminarily identified by Swiss staining and Giemsa staining,serological methods and molecular biological methods.On this basis,the micro-dilution method was used to determine the minimum inhibitory concentration (MIC) of the eight Chinese medicines against the isolated strains,in order to screen the Chinese medicines sensitive to the isolated strains.The results showed that after the isolated strains were proliferated in the liquid medium of Mycoplasma gallisepticum,the color of the medium changed from red to yellow and was translucent,after cultured in the solid medium,it showed a typical ‘fried egg’-like colony,which were consistent with the culture characteristics of Mycoplasma gallisepticum.The Swiss staining and Giemsa staining results indicated that the morphology of the bacteria was consistent with the characteristics of Mycoplasma gallisepticum.The serological identification results showed that the isolate had agglutinated with the positive serum of Mycoplasma gallisepticum.The molecular biology identification results showed the matching degree of the amplified nucleic acid sequence with Mycoplasma gallisepticum was as high as 99%.The result of MIC showed that the antibacterial activity of traditional Chinese medicines Coptis chinensis and Corcagium were sensitive to the isolated Mycoplasma gallisepticum,the MIC were ≤0.98 and 0.39 mg/mL,respectively.While the traditional Chinese medicine Fagopyrum dibotrys (D.Don) Hara and Houttuynia aucuparia were moderately sensitive to Mycoplasma gallisepticum,with MIC ≥125 mg/mL.Isatidis Radix,Nova cortices album,Angelica sinensis and Et persequebantur Josue were not sensitive to Mycoplasma gallisepticum.In conclusion,a Mycoplasma gallisepticum strain was successfully isolated in this study,and screened out four kinds of sensitive traditional Chinese medicines to this strain,which were Coptis chinesis,Corcagium,Fagopyrum dibotrys (D.Don) Hara and Houttuynia aucuparia,respectively,it provided a reference basis for further study of traditional Chinese medicine prescriptions of prevention and treatment of Mycoplasma gallisepticum.

Key words: Mycoplasma gallisepticum; isolation; traditional Chinese medicine; minimum inhibitory concentration

CLC Number:

S852.62

GUO Yangli, SONG Xiang, LIU Naizhi, QI Xiuye, CHENG Fuliang, XU Haiyan, GU Wei. Isolation and Identification of One Strain of Mycoplasma gallisepticum and Screening of Sensitive Traditional Chinese Medicine in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 1064-1071.

References

[1] 王育伟,岳华,张晓晖,等.鸡毒支原体研究概况[J].动物医学进展,2016,37(7):106-110. WANG Y W,YUE H,ZHANG X J,et al.The situation of research on Mycoplasma gallisepticum[J].Progress in Veterinary Medicine,2016,37(7):106-110.(in Chinese)
[2] GERCHMAN I,LEVISOHN S,MIKULA I,et al.Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticum strains isolated from poultry[J].Veterinary Research,2011,42(1):90.
[3] ISHFAQ M,HU W Y,KHAN M Z,et al.Current status of vaccine research,development,and challenges of vaccines for Mycoplasma gallisepticum[J].Poultry Science,2020,99(9):4195-4202.
[4] 张静,邵永斌,谷新利,等.紫花地丁总黄酮体外抗炎活性研究[J].中国畜牧兽医,2020,47(4):1258-1266. ZHANG J,SHAO Y B,GU X L,et al.Study on anti-inflammatory activity of total flavonoids from Viola yedoensis in vitro[J].China Animal Husbandry & Veterinary Medicine,2020,47(4):1258-1266.(in Chinese)
[5] 阎力君,吕欣彧,张多,等.黄连解毒汤有效成分的提取及其对体内白色念珠菌的清除作用[J].中国畜牧兽医,2019,46(11):3414-3421. YAN L J,LYU X Y,ZHANG D,et al.Extraction of effective fraction of Huanglian Jiedu decoction and its scavenging effect on Candida albicans in vivo[J].China Animal Husbandry & Veterinary Medicine,2019,46(11):3414-3421.(in Chinese)
[6] 刘云,朱善元,龚祝南,等.龙胆草及其提取物体外抗猪繁殖与呼吸综合征病毒作用的研究[J].中国畜牧兽医,2019,46(11):3449-3456. LIU Y,ZHU S Y,GONG Z N,et al.Study on the anti-procine reproductive and respiratory syndrome virus effect of gentian and its extract[J].China Animal Husbandry & Veterinary Medicine,2019,46(11):3449-3456.(in Chinese)
[7] 谭娅,甘麦邻,范源,等.金荞麦对脂多糖诱导小鼠小肠炎症的保护作用[J].中国畜牧兽医,2020,47(2):597-604. TAN Y,GAN M L,FAN Y,et al.Protective effects of Fagopyrum dibotrys on lipopolysaccharide-induced intestinal inflammation in mice[J].China Animal Husbandry & Veterinary Medicine,2020,47(2):597-604.(in Chinese)
[8] ZHANG Y H,REN L M,WANG X Y.Inhibitory effect of Houttuynia cordata Thunb on LPS-induced retinal microglial activation[J].International Journal of Ophthalmology,2019,12(7):1095-1100.
[9] 孟冬霞,贺东昌,武果桃,等.超微粉中药对鸡毒支原体的体外抑菌试验[J].畜牧兽医科技信息,2011,1:21-22. MENG D X,HE D C,WU G T,et al.Bacteriostatic test of ultrafine powder Chinese medicine on Mycoplasma gallisepticum in vitro[J].Chinese Journal of Animal Husbandry and Veterinary Medicine,2011,1:21-22.(in Chinese)
[10] 孟冬霞,贺东昌,武果桃,等.超微粉中药对人工感染鸡毒支原体病鸡的治疗试验[J].中国兽医杂志,2013,49(8):45-47. MENG D X,HE D C,WU G T,et al.Therapeutic test of ultrafine powder Chinese medicine on chickens infected with Mycoplasma gallisepticum[J].Chinese Journal of Veterinary Medicine,2013,49(8):45-47.(in Chinese)
[11] 李继昌,石乔,潘晓影,等.慢呼抗口服液对人工感染鸡毒支原体的治疗试验[J].中国家禽,2008,30(21):24-26. LI J C,SHI Q,PAN X Y,et al.The therapeutic test of Manhukang oral liquid on artificial infection of Mycoplasma gallisepticum[J].China Poultry,2008,30(21):24-26.(in Chinese)
[12] 郭如彬,沈明华,顾文松,等.中药复方提取物对人工诱发鸡毒支原体与大肠杆菌混合感染的疗效试验[J].中国家禽,2010,32(9):63-65. GUO R B,SHEN M H,GU W S,et al.Efficacy of compound extracts of traditional Chinese medicine on artificially induced mixed infection of Mycoplasma gallisepticum and Escherichia coli[J].China Poultry,2010,32(9):63-65.(in Chinese)
[13] 刘玲,管凤霞,黎敏,等.二株禽支原体的分离与鉴定[J].山东畜牧兽医,2010,31(4):17-18. LIU L,GUAN F X,LI M,et al.Isolation and identification of two avian Mycoplasma[J].Shandong Journal of Animal Science and Veterinary Medicine,2010,31(4):17-18.(in Chinese)
[14] 吴清民,杨秀玉,沈志强,等.鸡毒支原体的分离鉴定和最低抑菌浓度测定[J].中国预防兽医学报,2003,25(4):71-74. WU Q M,YANG X Y,SHENG Z Q,et al.Isolation and identification of Mycoplasma gallisepticum and determination of minimum inhibitory concentration[J].Chinese Journal of Preventive Veterinary Medicine,2003,25(4):71-74.(in Chinese)
[15] 陈美安,甄汉深,裴渭静,等.抗畜禽支原体中草药筛选及细胞毒性研究[J].动物医学进展,2014,35(4):33-38. CHEN M A,ZHEN H S,PEI W J,et al.Screening of Chinese herbal medicines against Mycoplasma in livestock and poultry and study on cytotoxicity[J].Progress in Veterinary Medicine,2014,35(4):33-38.(in Chinese)
[16] HU W Y,ZHANG W,SHAH S W A,et al.Mycoplasma gallisepticum infection triggered histopathological changes,oxidative stress and apoptosis in chicken thymus and spleen[J].Developmental & Comparative Immunology,2021,114:103832.
[17] LI P,XU J,HAO H M,et al.Mechanism of apoptosis induction by mycoplasmal nuclease MGA_0676 in chicken embryo fibroblasts[J].Frontiers in Cellular and Infection Microbiology,2018,8:105
[18] YUAN B,ZOU M Y,ZHAO Y B,et al.Up-regulation of miR-130b-3p activates the PTEN/PI3K/AKT/NF-κB pathway to defense against Mycoplasma gallisepticum (HS strain) infection of chicken[J].International Journal of Molecular Sciences,2018,19(8):2172.
[19] KATINKA B,ZSUZSA K,KINGA M S,et al.Genotyping Mycoplasma gallisepticum by multilocus sequence typing[J].Veterinary Microbiology,2019,231:191-196.
[20] KINGA M S,ZSUZSA K,KATINKA B,et al.Development of molecular methods for rapid differentiation of Mycoplasma gallisepticum vaccine strains from field isolates[J].Journal of Clinical Microbiology,2019,57(6):e01084-18.
[21] MARWA I,NAGLAA F S,YOUSREYA M,et al.In vitro evaluation of various antimicrobials against field Mycoplasma gallisepticum and Mycoplasma synoviae isolates in Egypt[J].Poultry Science,2019,98(12):6281-6288.
[22] 杨明彩,徐斌,隋兆峰,等.鸡毒支原体敏感的中草药筛选试验[J].中国家禽,2016,38(21):65-67. YANG M C,XU B,SUI Z F,et al.Screening of Chinese herbal medicines sensitive to Mycoplasma gallisepticum[J].China Poultry,2016,38(21):65-67.(in Chinese)
[23] 刘士学.中兽药治疗鸡毒支原体病的优势[J].畜牧兽医科技信息,2019,11:153-154. LIU S X.Advantages of Chinese veterinary medicine in the treatment of Mycoplasma gallisepticum[J].Chinese Journal of Animal Husbandry and Veterinary Medicine,2019,11:153-154.(in Chinese)
[24] MENG Y L,HUO J H,LU W H,et al.Modulation of p1 and EGF expression by Baicalin[J].Molecular Sciences,2013,14(1):146-157.
[25] 瞿瑜萍,何随彬,金耀忠,等.中兽药治疗鸡毒支原体病研究进展[J].上海畜牧兽医通讯,2017,3:19-21. QU Y P,HE S B,JIN Y Z,et al.Research progress on treatment of Mycoplasma gallisepticum with Chinese veterinary drugs[J].Shanghai Journal of Animal Husbandry and Veterinary Medicine,2017,3:19-21.(in Chinese)
[26] BAO J,WU Z,ISHFAQ M,et al.Comparison of experimental infection of normal and immuno-suppressed chickens with Mycoplasma gallisepticum[J].Journal of Comparative Pathology,2020,175:5-12.

Isolation and Identification of One Strain of Mycoplasma gallisepticum and Screening of Sensitive Traditional Chinese Medicine in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LU Yukui, LIU Jiaqi, ZHONG Jiacheng, CHEN Jidang, ZHU Wanjun, ZHANG Yishan, ZHANG Jipei. Isolation,Identification and Pathogenicity Analysis of Avibacterium paragallinarum from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2562-2572.
[2]	JIANG Shanyi, ZHOU Bijun, WEN Ming, WANG Kaigong. Prevention of Traditional Chinese Medicine with Probiotics on Eimeria tenella [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2092-2102.
[3]	CHEN Tianbao, WANG Yani, LIU Baoling, WANG Gang, WANG Xiaohu, LIU Xuhui, CAI Rujian, HE Dongsheng. Research Progress on the Anti-Porcine Reproductive and Respiratory Syndrome Virus Effect and Mechanism of Traditional Chinese Medicine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2114-2129.
[4]	LI Zhijie, ZHAO Ying, MA Xin, WANG Meng, YU Sijiu, WANG Libin, PAN Yangyang. Research Progress on Isolation Methods and Main Reproductive Functions of Exosomes from Animal Follicular Fluid [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1480-1488.
[5]	TIAN Changqing, ZHANG Kunzhong, QI Yumei, SHI Wenjing, DONG Zhijie, ZHANG Haohao, HE Zengwen, ZHI Ji, ZHAO Xuehui, CHONG Qian, XUE Huiwen, GOU Huitian. Isolation,Identification and Biological Characteristics Analysis of a Strain of Listeria monocytogenes from Environment [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1543-1555.
[6]	CAO Yingying, LIANG Liang, ZENG Yirong, ZHONG Qinghua, YUAN Xiaofang, TANG Haibo, LENG Jing. Isolation,Identification and Biological Characteristics of Salmonella from Tupaia belangeri chinensis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1718-1728.
[7]	LIU Kexin, WANG Chao, ZHANG Ao, LIU Jiali, TAN Bin, ZHANG Shuqin. Isolation,Identification and Genome Sequence Analysis of Bovine Viral Diarrhea Virus LN-1 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1150-1159.
[8]	CHEN Dong, ZHANG Yanhao, ZHANG Yang, LI Sen, ZHOU Zutao, LI Zili. Isolation,Identification and Drug Sensitivity of 3 Strains of Riemerella anatipestifer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1250-1258.
[9]	WANG Bingyi, MA Hongcai, ZOU Minghao, FAN Shijie, YUAN Zhenjie, Labaciren, DONG Hailong, ZENG Jiangyong. Isolation,Identification and Drug Resistance Analysis of Streptococcus from Yak [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 745-753.
[10]	WANG Wei, LI Chunzhu, WANG Shasha, SANG Rui, GE Bingjie, LI Chunting, ZHAO Xin, LIU Xinman, YU Minghong, ZHANG Xuemei. Study on the Protective Mechanism of Traditional Chinese Medicine Combined with Probiotics on Diarrhea Caused by E.coli in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 779-788.
[11]	ZHONG Huachen, WANG Lifang, GUO Chenyang, LIU Jialin, SONG Jie. Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 817-826.
[12]	SUN Yu, ZHANG Hao, LI Changyang, TAO Yanzi, WANG Qiuju. Isolation,Identification and Biological Characteristics of Antioxidant Bacillus from Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 359-367.
[13]	WEI Jin, WANG Jia, ZHAO Haoran, LIU Bo, LIU Yuanjie, YAO Wensheng, LIU Yan, MA Xin. Isolation, Identification and Antimicrobial Susceptibility of Mycoplasma gallisepticum from Different Areas [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 368-376.
[14]	LIU Hongying, WANG Jingyu, CUI Zhenzhen, MA Qingxia, WANG Tingting, LI Jianming, SHI Kun, GONG Qinglong, DU Rui, LENG Xue. Research Progress on Immune Adjuvant Effect and Related Mechanism of Active Ingredients of Traditional Chinese Medicine on Animal Vaccines [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 390-397.
[15]	YANG Fumei, YANG Linfu, ZENG Cheng, YANG Zhi, WANG Kang, DUAN Gang, DAI Feiyan. Isolation,Identification and Biological Characterization Analysis of a Strain of Carnobacterium maltaromaticum from Barking Deer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 398-407.