连花清瘟药渣及发酵产物的生物活性研究

doi:10.11843/j.issn.0366-6964.2023.03.039

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (3): 1281-1299.doi: 10.11843/j.issn.0366-6964.2023.03.039

连花清瘟药渣及发酵产物的生物活性研究

刘广^1,2, 龚成燕¹, 牛凯敏¹, 张硕³, 陶灯³, 马军⁴, 吴信¹, 印遇龙¹, 唐宇龙^1*

1. 中国科学院亚热带农业生态研究所中国科学院亚热带农业生态过程重点实验室湖南省畜禽健康养殖工程技术中心农业部中南动物营养与饲料科学观测实验站, 长沙 410125;
2. 湖南农业大学动物科学与技术学院, 长沙 410125;
3. 云南西南农牧集团股份有限公司, 昆明 650217;
4. 石家庄以岭药业股份有限公司, 石家庄 050035

收稿日期:2022-03-02 出版日期:2023-03-23 发布日期:2023-03-21
通讯作者: 唐宇龙,主要从事生猪健康养殖研究,E-mail:tangyulong@isa.ac.cn
作者简介:刘广(1998-)，男，湖南益阳人，硕士生，主要从事动物营养与饲料科学研究，E-mail:lg19980905@163.com
基金资助:
湖南省重点研发项目(2022NK2054)；云南省重大科技专项(202102AE090046)；湖南省科技领军人才专项(2019RS3021；2019RS3020)

Study on Biological Activity of the Residue and Fermentation Products of Lianhua Qingwen

LIU Guang^1,2, GONG Chengyan¹, NIU Kaimin¹, ZHANG Shuo³, TAO Deng³, MA Jun⁴, WU Xin¹, YIN Yulong¹, TANG Yulong^1*

1. Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central of Ministry of Agriculture, Hunan Provincial Engineering and Technology Center for Healthy Livestock and Poultry Production, Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;
2. College of Animal Science and Technology, Hunan Agricultural University, Changsha 410125, China;
3. Yunnan Southwest Agriculture and Animal Husbandry Group Co. LTD., Kunming 650217, China;
4. Shijiazhuang Yiling Pharmaceutical Co. Ltd., Shijiazhuang 050035, China

Received:2022-03-02 Online:2023-03-23 Published:2023-03-21

摘要/Abstract

摘要： 旨在研究连花清瘟药渣和发酵产物的生物学活性，探明自然发酵、优化发酵1(4MYL)和优化发酵2(Y4ML)三种发酵方式对药渣中营养成分的改变以及乙醇提取物和水提取物抑菌效果及抗病毒效果的差异。采用肉汤稀释法测定药渣发酵前后不同提取物对猪副嗜血杆菌(Haemophilus parasuis，HPS)、猪链球菌二型(Streptococcus suis type 2，SS2)、肠毒性大肠杆菌(Enterotoxigenic Escherichia coli，ETEC)3种致病菌菌株的最小抑菌浓度(MIC)；以伪狂犬病病毒(Pseudorabies virus，PRV)、猪圆环病毒2型(porcine circovirus type 2，PCV2)、猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus, PRRSV)分别感染非洲绿猴肾细胞(Vero)、非洲绿猴胚胎肾细胞(MARC-145)、猪肾细胞(PK-15)，通过荧光显微镜观察细胞形态病变，同时结合实时荧光定量PCR技术进行病毒核酸检测和定量作为药物体外抗病毒效果的评价指标。结果表明，连花清瘟原药渣的乙醇和水提取物对HPS和SS2均表现出抑制作用，但对ETEC无抑制作用；对3种试验病毒均表现出抗病毒活性，但抗病毒活性存在一定区别。发酵后乙醇和水提取物增强了抗菌抗病毒活性，同时对ETEC表现出抑制作用。综上表明，连花清瘟药渣提取物具有不同程度的抗菌、抗病毒作用，能有效抑制猪养殖过程中HPS、SS2等常见的致病菌和病毒，发酵后能增强抗菌和抗病毒效果，具有开发成动物饲料或饲料添加剂的潜力。

关键词: 连花清瘟药渣, 发酵, 抗菌, 抗病毒

Abstract: The purpose of this experiment was to study the biological activities of Lianhua Qingwen residue and its fermentation products, and to investigate the differences among the methods of fermentation including natural, optimised fermentation 1 (4MYL) and optimised fermentation 2 (Y4ML) in terms of the alteration of the nutrient content of the residue and the antibacterial and antiviral effects of the ethanolic and aqueous extracts. Broth dilution method was used to determine the minimum inhibitory concentration (MIC) of different extracts of drug residue before and after fermentation against three pathogenic strains of Haemophilus parasuis (HPS), Streptococcus suis type 2 (SS2) and Enterotoxic Escherichia coli (ETEC); African green monkey kidney cells (Vero), African green monkey embryonic kidney cells (MARC-145) and porcine kidney cells (PK-15) were infected with pseudorabies virus (PRV), porcine circovirus type 2 infection (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), respectively. The morphological changes of cells were observed by fluorescence microscope, and the virus nucleic acid was detected and quantified by real-time fluorescence quantitative PCR as an evaluation index of the antiviral effect of drugs in vitro. The results showed that the ethanolic and aqueous extracts of Lianhua Qingwen medicinal dregs showed inhibitory effects on HPS and SS2 strains, but had no inhibitory effect on ETEC; They all showed antiviral activity against the three test viruses, but there existed certain differences in antiviral activity. After fermentation, the ethanolic and aqueous extracts enhanced their antibacterial and antiviral activities and inhibited ETEC. It can be concluded that the extract of Lianhua Qingwen residue has different degrees of antibacterial and antiviral effects, can effectively inhibit HPS, SS2 and other common pathogenic bacteria and viruses in the process of pig breeding, and antibacterial and antiviral effect are enhanced after fermentation. The above results supported that Lianhua Qingwen residue extract has the potential to be developed into animal feed or feed additive.

Key words: Lianhua Qingwen residue, fermentation, antibacterial, antiviral

中图分类号:

S853.74

刘广, 龚成燕, 牛凯敏, 张硕, 陶灯, 马军, 吴信, 印遇龙, 唐宇龙. 连花清瘟药渣及发酵产物的生物活性研究[J]. 畜牧兽医学报, 2023, 54(3): 1281-1299.

LIU Guang, GONG Chengyan, NIU Kaimin, ZHANG Shuo, TAO Deng, MA Jun, WU Xin, YIN Yulong, TANG Yulong. Study on Biological Activity of the Residue and Fermentation Products of Lianhua Qingwen[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1281-1299.

参考文献 36

[1]	郭义东,何兴,冯兴,等.中药渣综合利用研究进展[J].成都大学学报(自然科学版),2015,34(2):125-128.GUO Y D,HE X,FENG X,et al.Research progress in comprehensive utilization of traditional Chinese medicine dregs[J].Journal of Chengdu University(Natural Science Edition),2015,34(2):125-128.(in Chinese)
[2]	赵志超,图门巴雅尔.中草药提取残渣综合利用研究进展[J].畜牧与饲料科学,2019,40(2):47-52.ZHAO Z C,TUMENBAYAER.Research progress on comprehensive utilization of extracting residues from traditional Chinese medicine[J].Animal Husbandry and Feed Science,2019,40(2):47-52.(in Chinese)
[3]	刘洋,金顺义,常娟,等.复合益生菌发酵中草药前后活性成分变化[J].安徽农业科学,2017,45(34):123-125.LIU Y,JIN S Y,CHANG J,et al.Changes of active ingredients before and after compound probiotic fermented Chinese herbs[J].Journal of Anhui Agricultural Sciences,2017,45(34):123-125.(in Chinese)
[4]	张文意,张婧,高鹏飞,等.槐耳/板蓝根双向发酵体系物质基础变化[J].中国实验方剂学杂志,2016,22(16):66-70.ZHANG W Y,ZHANG J,GAO P F,et al.Changes in chemical component content of fungal substance Trametes robiniophila/isatidis radix co-fermentation system[J].Chinese Journal of Experimental Traditional Medical Formulae,2016,22(16):66-70.(in Chinese)
[5]	陈旸,王义,孙亮,等.植物乳杆菌发酵转化人参皂苷的研究[J].中国中药杂志,2014,39(8):1435-1440.CHEN Y,WANG Y,SUN L,et al.Fermentation transformed ginsenoside by Lactobacillus plantarum[J].China Journal of Chinese Materia Medica,2014,39(8):1435-1440.(in Chinese)
[6]	戴燊,程茂基,韩信,等.中草药混菌发酵生产新型生物中药的研究[J].饲料工业,2013,34(11):20-24.DAI S,CHENG M J,HAN X,et al.Research on the fermentation of Chinese herbal medicine with mixed bacteria to produce new biological traditional Chinese medicine[J].Feed Industry,2013,34(11):20-24.(in Chinese)
[7]	苏建树,刘白宁,田平芳,等.微生物发酵对川乌、附子中生物碱含量的影响[J].北京化工大学学报(自然科学版),2010,37(3):97-101.SU J S,LIU B N,TIAN P F,et al.Effect of microbial fermentation on the extraction of alkaloids from radix aconiti and aconite[J].Journal of Beijing University of Chemical Technology(Natural Science Edition),2010,37(3):97-101.(in Chinese)
[8]	苏贵龙.益生菌FGM发酵对黄芪根、茎、叶主要活性成分含量的影响研究[D].北京:中国农业科学院,2017.SU G L.The effect of fermentation with FGM strain on the main active ingredients in Astragalus root,Stem and Leaf[D].Beijing:Chinese Academy of Agricultural Sciences,2017.(in Chinese)
[9]	丁保安,李转见,莫元春.红景天药渣对肉鸡生长性能、营养物质代谢和血液指标的影响[J].中国家禽,2010,32(3):16-19.DING B A,LI Z J,MO Y C.Effect of dietary red common sonecrop herb residues on growth performance,nutrient metabolism and blood index of broilers[J].China Poultry,2010,32(3):16-19.(in Chinese)
[10]	张恬,李娜,王多伽,等.黄芪药渣在松辽黑猪生产中的应用研究[J].饲料与畜牧,2019(11):36-38.ZHANG T,LI N,WANG D J,et al.Application of Astragalus residue in Songliao black pig production[J].Animal Agriculture,2019(11):36-38.(in Chinese)
[11]	王恩花,蔡兴芳,张富林,等.降解黄芪药渣对肉鸡生产性能及风味的影响[J].贵州农业科学,2020,48(10):69-72.WANG E H,CAI X F,ZHANG F L,et al.Effects of degraded astragalus residue on growth performance and flavor of broilers[J].Guizhou Agricultural Sciences,2020,48(10):69-72.(in Chinese)
[12]	王英伟.水飞蓟残渣在育肥猪生产上的应用[D].延吉:延边大学,2006.WANG Y W.Application of milk thistle residue in Finishing pig production[D].Yanji:Yanbian University,2006.(in Chinese)
[13]	钟莲.饲料中添加中草药"催奶灵"散提高牛奶产量及质量试验[J].中兽医学杂志,2016(4):20.ZHONG L.Experimental study on improving milk yield and quality by adding Chinese herbal medicine "rinnai ling" powder to feed[J].Chinese Journal of Traditional Veterinary Science,2016(4):20.(in Chinese)
[14]	李瑞娟,梁娜,冯芳,等.复方中草药饲料添加剂对仔猪断奶应激的临床效果试验[J].今日畜牧兽医,2021,37(10):17-18.LI R J,LIANG N,FENG F,et al.Clinical effect of compound Chinese herbal feed Additive on weaning stress of piglets[J].Today Animal Husbandry and Veterinary Medicine,201,37(10):17-18.(in Chinese)
[15]	李雪男,李梦霄,魏佳荣,等.中草药饲料添加剂对蛋鸡生产性能及蛋品质的影响[J].北方牧业,2021(19):22.LI X N,LI M X,WEI J R,et al.Effects of Chinese Herbal feed additives on performance and egg quality of Laying Hens[J].Northern Animal Husbandry,2021(19):22.(in Chinese)
[16]	姜露,郭文韬,陈志明,等.中草药饲料添加剂对育肥猪免疫功能、抗氧化能力以及生长激素水平的影响[J].中国兽医科学,2022,52(1):120-127.JIANG L,GUO W T,CHEN Z M,et al.Effects of Chinese herbal medicine feed additives on immune function,antioxidant capacity and growth hormone levels of finishing pigs[J].Chinese Veterinary Science,2002,52(1):120-127.(in Chinese)
[17]	苏美,吕武兴,邓敦.中草药饲料添加剂增强家禽抗病力的作用和应用[J].湖南饲料,2021(3):27-30.SU M,LV W X,DENG D.Effect and application of Chinese herbal feed additives to enhance disease resistance of poultry[J].Hunan Feed,2021(3):27-30.(in Chinese)
[18]	邹士海,吴颖.中草药添加剂对母猪及仔猪免疫力的影响[J].浙江畜牧兽医,2021,46(4):1-3.ZOU S H,WU Y.Effects of Chinese herbal additives on Immunity of Sows and Piglets[J].Zhejiang Journal Animal Science and Veterinary Medicine,2021,46(4):1-3.(in Chinese)
[19]	朱从磊,胡芳英.莲花清瘟用于病毒感染性疾病的文献分析[J].宜春学院学报,2020,42(12):31-36,82.ZHU C L,HU F Y.Literature review on the application of Lianhuaqingwen in viral infectious diseases[J].Journal of Yichun University,2020,42(12):31-36,82.(in Chinese)
[20]	尹玉洁,常丽萍.中药连花清瘟胶囊/颗粒在呼吸系统疾病中的药理研究及临床应用进展[J].中国临床药理学与治疗学,2021,26(10):1174-1180.YIN Y J,CHANG L P.Pharmacological research and clinical applications progress of Lianhua Qingwen capsules/granules in respiratory diseases[J].Chinese Journal of Clinical Pharmacology and Therapeutics,2021,26(10):1174-1180.(in Chinese)
[21]	叶春娟,张帆,朱育银.莲花清瘟胶囊对流行性感冒合并支气管肺炎的作用[J].中国实验方剂学杂志,2013,19(24):329-331.YE C J,ZHANG F,ZHU Y Y.Clinical study of Lianhua Qingwen capsule in treatment of influenza combined with bronchial pneumonia[J].Chinese Journal of Experimental Traditional Medical Formulae,2013,19(24):329-331.(in Chinese)
[22]	樊拖迎,肖明生,周虎.莲花清瘟胶囊联合阿奇霉素治疗高龄患者急性细菌性上呼吸道感染的临床研究[J].现代诊断与治疗,2019,30(6):856-857.FAN T Y,XIAO M S,ZHOU H.Clinical study of Lianhua Qingwen Capsule combined with azithromycin in treatment of acute bacterial upper respiratory tract infection in elderly patients[J].Modern Diagnosis and Treatment,2019,30(6):856-857.(in Chinese)
[23]	TANG Y L,ZHANG X Y,WU W,et al.Inactivation of the sodA gene of Streptococcus suis type 2 encoding superoxide dismutase leads to reduced virulence to mice[J].Vet Microbiol,2012,158(3-4):360-366.
[24]	TANG Y L,LI F N,TAN B,et al.Enterotoxigenic Escherichia coli infection induces intestinal epithelial cell autophagy[J].Vet Microbiol,2014,171(1-2):160-164.
[25]	ZHANG Q,LIU J T,YAN S X,et al.Fluoroquinolone-resistant Haemophilus parasuis isolates exhibit more putative virulence factors than their susceptible counterparts[J].J Clin Microbiol,2013,51(9):3130-3131.
[26]	ZHAN Y,ZHANG L H,LIN Y,et al.Development and preliminary testing of a probe-based duplex real-time PCR assay for the detection of African swine fever virus[J].Mol Cell Probes,2021,59:101764.
[27]	王林山,尹燕博,邹勇,等.PCR和IFA测定猪圆环病毒2型半数细胞培养感染量的比较[J].上海农业学报,2013,29(1):34-38.WANG L S,YIN Y B,ZOU Y,et al.Comparison between PCR and indirect immunofluorescence assay (IFA) methods for determining PCV2 TCID50[J].Acta Agriculturae Shanghai,2013,29(1):34-38.(in Chinese)
[28]	潘文,马良,张贺楠,等.猪伪狂犬病病毒ZY-2014株的分离与鉴定[J].中国兽药杂志,2016,50(5):1-6.PAN W,MA L,ZHANG H N,et al.Isolation and identification of variant strain ZY-2014 of pseudorabies virus[J].Chinese Journal of Veterinary Drug,2016,50(5):1-6.(in Chinese)
[29]	段金廒,郭盛,唐志书,等.中药资源循环利用模式构建及产业化示范[J].江苏中医药,2019,51(3):1-5.DUAN J A,GUO S,TANG Z S,et al.The construction and industrialization demonstration of recycling model of Chinese medicinal resources[J].Jiangsu Journal of Traditional Chinese Medicine,2019,51(3):1-5.(in Chinese)
[30]	孙晓燕.微生物快速发酵中药渣制备无抗饲料添加剂及其应用效果的研究[D].广州:华南理工大学,2019.SUN X Y.Study on the preparation of anti-feed additive and its application effect of microorganism rapid fermentation of Chinese medicine residue[D].Guangzhou:South China University of Technology,2019.(in Chinese)
[31]	宋玉琴.人参药渣添加到动物饲料调节动物抵抗力和肉质的实验研究[D].成都:成都中医药大学,2016.SONG Y Q.Study of ginseng residue added to animal feeds on regulating resistance and meat quality for animals[D].Chengdu:Chengdu University of Traditional Chinese Medicine,2016.(in Chinese)
[32]	陈鑫,李芳蓉,张建军,等.红芪药渣固态发酵生产蛋白饲料工艺条件优化[J].中国酿造,2019,38(6):176-180.CHEN X,LI F R,ZHANG J J,et al.Technological conditions optimization of protein feed production from Radix hedysari residues by solid-state fermentation[J].China Brewing,2019,38(6):176-180.(in Chinese)
[33]	简娟娟,刘利,王维.猪链球菌2型感染的临床特征和治疗[J].现代畜牧科技,2015(2):69.JIAN J J,LIU L,WANG W.Clinical characteristics and treatment of Streptococcus suis type 2 infection[J].Modern Animal Husbandry Science&Technology,2015(2):69.(in Chinese)
[34]	张斌,鲍惠玲,刘洋.猪副嗜血杆菌病的流行病学、诊断和防控措施[J].现代畜牧科技,2019(5):115-116.ZHANG B,BAO H L,LIU Y.Epidemiology,diagnosis and control measures of Haemophilus parasubiosis of porcine[J].Modern Animal Husbandry Science&Technology,2019(5):115-116.(in Chinese)
[35]	MIRHOSEINI A,AMANI J,NAZARIAN S.Review on pathogenicity mechanism of enterotoxigenic Escherichia coli and vaccines against it[J].Microb Pathog,2018,117:162-169.
[36]	陈溥言.兽医传染病学[M].6版.北京:中国农业出版社,2015.CHEN P Y.Veterinary infectious diseases[M].6th ed.Beijing:China Agriculture Press,2015.(in Chinese)

连花清瘟药渣及发酵产物的生物活性研究

Study on Biological Activity of the Residue and Fermentation Products of Lianhua Qingwen

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 36

相关文章 15

编辑推荐

Metrics

本文评价

[1]	陈姝宇, 朱雪蛟, 周金柱, 陶然, 张雪寒, 索朗斯珠, 贡嘎, 李彬. 鸟苷酸结合蛋白GBP1和GBP2抑制猪轮状病毒体外复制[J]. 畜牧兽医学报, 2024, 55(1): 245-257.
[2]	王姿月, 张子卉, 吴文学, 彭辰. 猴痘的诊断、预防和治疗[J]. 畜牧兽医学报, 2023, 54(8): 3195-3205.
[3]	熊程坤, 张道亮, 杨悦, 丁红研, 赵杰, 李玉, 王希春, 冯士彬, 赵畅, 汤继顺, 吴金节. 芦丁对围产期湖羊瘤胃发酵、瘤胃菌群结构及抗氧化性能的影响[J]. 畜牧兽医学报, 2023, 54(7): 2898-2909.
[4]	李倬伟, 王方, 王君君, 陈祯涵, 李焕荣, 周双海, 刘雪威. 双香豆素对猪繁殖与呼吸综合征病毒的体外抑制作用[J]. 畜牧兽医学报, 2023, 54(3): 1160-1168.
[5]	卓儒浩, 柳清扬, 钟翔. 姜黄素调控肠道菌群及抗病毒作用研究进展[J]. 畜牧兽医学报, 2023, 54(2): 473-483.
[6]	孙敏, 郝飞, 张纹纹, 李文良, 杨蕾蕾, 毛立, 程子龙, 刘茂军. IFN-α对山羊副流感病毒3型的抗病毒活性[J]. 畜牧兽医学报, 2023, 54(2): 736-743.
[7]	宫浩阳, 吴佳鑫, 杨晓钰, 解伟纯, 王雪莹, 李佳璇, 姜艳平, 崔文, 李一经, 唐丽杰. 肠道菌群抗病毒机制研究进展[J]. 畜牧兽医学报, 2023, 54(12): 4910-4919.
[8]	范锦全, 张宇航, 唐午阳, 赵欣宇, 李丕顺, 郑晓峰. 地西他滨对猪圆环病毒2型的体外抑制作用[J]. 畜牧兽医学报, 2023, 54(12): 5134-5142.
[9]	任红阳, 郑林峰, 王自立, 王红亮. 混贮原料对全株玉米青贮粗蛋白质含量及发酵品质的影响[J]. 畜牧兽医学报, 2023, 54(11): 4526-4536.
[10]	陈文哲, 张向乐, 顾峰幸, 赵振翔, 李康丽, 薛钊宁, 郑海学, 张小丽, 朱紫祥. 猪GRK2蛋白抗口蹄疫病毒作用分析[J]. 畜牧兽医学报, 2023, 54(10): 4350-4361.
[11]	武周慧, 王瑜, 杜衡, 王之文, 肖爽, 武金亮, 王真. 替拉扎明对多重耐药沙门菌抗菌增敏活性分析[J]. 畜牧兽医学报, 2023, 54(10): 4362-4371.
[12]	常伟辰, 李帅奇, 李琰, 闫微, 张红英, 王彦彬, 杨明凡, 张昂克. 白头翁散煎剂发酵物对感染猪流行性腹泻病毒仔猪肠道屏障功能的影响[J]. 畜牧兽医学报, 2023, 54(10): 4403-4410.
[13]	韦肖, 张建童, 龙唐晖, 李开嵘, 李艳娇, 欧阳克蕙, 邱清华. 日粮能量水平对湖羊瘤胃发酵特性和微生物组成的影响[J]. 畜牧兽医学报, 2022, 53(9): 3042-3051.
[14]	孙栋, 屈莎, 李璇, 唐善虎, 李思宁, 郝刚. 抗菌肽Tachyplesin Ⅰ及衍生物对大肠杆菌细胞膜作用机制的对比研究[J]. 畜牧兽医学报, 2022, 53(9): 3180-3189.
[15]	姚晨, 郭萌, 胡慧, 史晨曦, 杨国宇. 苯基吡啶酮衍生物影响猪δ冠状病毒复制的作用分析[J]. 畜牧兽医学报, 2022, 53(9): 3190-3198.