24味中草药与抗生素联合对2种病原菌的抑菌作用研究

doi:10.16431/j.cnki.1671-7236.2025.07.039

摘要/Abstract

摘要： 【目的】研究常用抗生素和24味中草药对嗜水气单胞菌(Aeromonas hydrophila)和蜡样芽孢杆菌(Bacillus cereus)的抑菌作用，以及联合用药的抑菌效果，筛选出有效抗菌组方用于生产实践，治疗细菌病感染。【方法】采用药敏纸片法筛选出对嗜水气单胞菌和蜡样芽孢杆菌抑制作用较强的几种抗生素，并测定这几种抗生素和24味中草药对上述2种病原菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。根据中草药的MIC和MBC筛选出有效的中草药分别与抗生素进行配伍联用，采用微量肉汤棋盘稀释法测定联用药物配对对嗜水气单胞菌和蜡样芽孢杆菌的分级抑菌浓度指数(fractional inhibitory concentrations index,FICI)，判断药物之间的联合使用效果，筛选出有效抑菌的中西药联合，并将其中1组中西药制成联合方剂进行体内试验。【结果】单独使用抗生素和中草药对嗜水气单胞菌的抑菌效果显示，抗生素恩诺沙星效果最佳(MIC为0.39 mg/mL)，中草药黄芩效果最佳(MIC为7.81 mg/mL)；联合用药结果显示，黄芩与恩诺沙星的FICI为0.127，呈协同作用，石榴皮与恩诺杀星的FICI为0.562，呈相加作用。单独使用抗生素和中草药对蜡样芽孢杆菌的抑菌效果显示，抗生素头孢他啶效果最佳(MIC为0.78 mg/mL)，中草药黄连和黄芩效果最佳(MIC均为7.81 mg/mL)；联合用药结果显示，黄连与头孢他啶的FICI为0.5，呈协同作用。联合用药体内试验结果显示，石榴皮和恩诺沙星联合用药组大口黑鲈肝脏结构完整，肝细胞排列紧密，同时肠道各组织界限分明，肠黏膜完整，对肠道起到较好的保护作用，有效缓解了嗜水气单胞菌对大口黑鲈肝脏和肠道造成的损伤。【结论】在体外试验中，组方黄芩联合恩诺沙星和石榴皮联合恩诺沙星均对嗜水气单胞菌有抑制作用；组方黄连联合头孢他啶对蜡样芽孢杆菌有较强的抑菌效果。石榴皮和恩诺沙星联合用药对嗜水气单胞菌感染的大口黑鲈有较好的治疗作用。

关键词: 中草药; 抗生素; 嗜水气单胞菌; 蜡样芽孢杆菌; 分级抑菌浓度指数(FICI)

Abstract: 【Objective】 The experiment aimed to study the bacteriostatic effect of commonly used antibiotics and 24 Chinese herbal medicines on Aeromonas hydrophila and Bacillus cereus,as well as the bacteriostatic effect of the combination of drugs,so as to select effective antibacterial formulas for production practice and treat bacterial infections. 【Method】 Several antibiotics with strong inhibitory effect on Aeromonas hydrophila and Bacillus cereus were screened by drug sensitive disk method,and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of these antibiotics and 24 Chinese herbal medicines against the above two pathogens were determined.According to the MIC and MBC of Chinese herbal medicine,the effective Chinese herbal medicine was selected to be combined with antibiotics.The fractional inhibitory concentrations index (FICI) of the combined drugs against Aeromonas hydrophila and Bacillus cereus was determined by the broth chessboard dilution method.The combined effect of drugs was judged,and the effective combination of Chinese and Western medicine was selected.One group of Chinese and Western medicine was made into a combined prescription for in vivo test. 【Result】 The antibacterial effect of antibiotics and Chinese herbal medicine on Aeromonas hydrophila showed that the antibiotic enrofloxacin had the best effect (MIC was 0.39 mg/mL),and the Chinese herbal medicine Scutellaria baicalensis Georgi had the best effect (MIC was 7.81 mg/mL).The results of combined medication showed that the FICI of Scutellaria baicalensis Georgi and enrofloxacin was 0.127,showing a synergistic effect,and the FICI of Pericarpium granati and enrofloxacin was 0.562,showing an additive effect.The antibacterial effect of antibiotics and Chinese herbal medicine on Bacillus cereus showed that the antibiotic ceftazidime had the best effect (MIC was 0.78 mg/mL),and the Chinese herbal medicine Coptis chinensis Franch.and Scutellaria baicalensis Georgi had the best effect (MIC both were 7.81 mg/mL).The results of combined medication showed that the FICI of Coptis chinensis Franch.and ceftazidime was 0.5,showing a synergistic effect.The in vivo test results of the combined medication showed that Pericarpium granati and enrofloxacin combined medication group had complete liver structure,closely arranged hepatocytes,at the same time,the intestinal tissues were clearly defined,and the intestinal mucosa was intact,and had a good protective effect on the intestinal mucosa,effectively alleviating the damage caused by Aeromonas hydrophila to the liver and intestine of Micropterus salmoides. 【Conclusion】 In vitro test,Scutellaria baicalensis Georgi combined with enrofloxacin and Pericarpium granati combined with enrofloxacin had inhibitory effect on Aeromonas hydrophila.Coptis chinensis Franch.combined with ceftazidime had strong antibacterial effect on Bacillus cereus.The combination of Pericarpium granati and enrofloxacin had a good therapeutic effect on Aeromonas hydrophila infected Micropterus salmoides.

Key words: Chinese herbal medicine; antibiotic; Aeromonas hydrophila; Bacillus cereus; fractional inhibitory concentration index (FICI)

中图分类号:

S853.7

黄春杏, 梁延达, 赖明建, 韦木莲, 杨映. 24味中草药与抗生素联合对2种病原菌的抑菌作用研究[J]. 中国畜牧兽医, 2025, 52(7): 3409-3419.

HUANG Chunxing, LIANG Yanda, LAI Mingjian, WEI Mulian, YANG Ying. Study on the Antibacterial Effect of 24 Chinese Herbs Medicines Combined with Antibiotics on Two Pathogenic Bacteria[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3409-3419.

参考文献

[1] 丁鹏.抗生素在养殖业中过度使用的危害及控制措施[J].北方牧业,2023，17:15.DING P.The dangers of overuse of antibiotics in agriculture and control measures[J].Beifang Muye,2023,17:15.(in Chinese)
[2] 胡建美,王宝屯,冯娟,等.60种中草药及其复方对杀鱼爱德华氏菌的体外抑菌效果[J].甘肃农业大学学报,2022,57(2):27-35.HU J M,WANG B T,FENG J,et al. In vitro bacteriostatic effect of 60 Chinese herbal medicines and their compounds on Edwardsiella piscicida[J].Journal of Gansu Agricultural University,2022,57(2):27-35.(in Chinese)
[3] 丁月霞,欧慧慧,王玉婷,等.中草药与抗生素联用对猪源沙门氏杆菌的体外抑菌效果观察[J].黑龙江畜牧兽医,2020,1:110-113.DING Y X,OU H H,WANG Y T,et al.Observation on the in vitro antibacterial effect of Chinese herbs combined with antibiotics against Salmonella from pigs[J].Heilongjiang Animal Science and Veterinary Medicine,2020,1:110-113.(in Chinese)
[4] 董亚萍.中草药延缓嗜水气单胞菌对恩诺沙星的耐药性及其作用机制的研究[D].上海：上海海洋大学,2018.DONG Y P.Study on Chinese herbal delaying Aeromonas hydrophila resistance on enrofloxacin and its mechanism[D].Shanghai:Shanghai Ocean University,2018.(in Chinese)
[5] 晋鹏飞,张浩然,王婷婷,等.5种中草药的不同溶剂提取物对嗜水气单胞菌抑菌活性的研究[J].水产科技情报,2022,49(4):212-218.JIN P F,ZHANG H R,WANG T T,et al.Study on antibacterial activity of different solvent extracts from five Chinese herbal medicines against Aeromonas hydrophila[J].Fisheries Science & Technology Information,2022,49(4):212-218.(in Chinese)
[6] 刘宇航.三种药物在抗草鱼食源性肠炎和嗜水气单胞菌感染防治中的机理研究[D].大连：大连海洋大学,2024.LIU Y H.Mechanism study of three drugs in the prevention and treatment of foodborne enteritis and Aeromonas hydrophila infection in grass carp[D].Dalian:Dalian Ocean University,2024.(in Chinese)
[7] 孟欣,孙大庆,张冬星,等.中草药提取物对东北林蛙源嗜水气单胞菌体内外抑菌试验及其抗炎免疫活性研究[J].黑龙江畜牧兽医,2022,14:112-118.MENG X,SUN D Q,ZHANG D X,et al.Antibacterial activity anti-inflammatory and immune activity of Chinese herbal extracts against Aeromonas hydrophila from Northeastern forest frogs in vivo and in vitro[J].Heilongjiang Animal Science and Veterinary Medicine,2022,14:112-118.(in Chinese)
[8] 李亚菲,张秀凤,张灵璇,等.应对细菌耐药策略研究进展[J].中国畜牧兽医,2023,50(11):4568-4576.LI Y F,ZHANG X F,ZHANG L X,et al.Research progress on strategies to control bacterial resistance:A review[J].China Animal Husbandry & Veterinary Medicine,2023,50(11):4568-4576.(in Chinese)
[9] 赵海晴.联合用药对嗜水气单胞菌耐药性影响与互作机制研究[D].北京：中国农业科学院,2023.ZHAO H Q.Study on the effect and interaction mechanism of combined drugs on drug resistance of Aeromonas hydrophila[D].Beijing:Chinese Academy of Agricultural Sciences,2023.(in Chinese)
[10] 邵明珠,赵允清,李松励,等.河北省奶牛源大肠杆菌的分离鉴定、致病性及中草药体外抑菌效果研究[J].中国畜牧兽医,2022,49(9):3683-3691.SHAO M Z,ZHAO Y Q,LI S L,et al.Isolation and identification of Escherichia coli from dairy cows in Hebei province and its pathogenicity and antibacterial effect of Chinese herbal medicine in vitro[J].China Animal Husbandry & Veterinary Medicine,2022,49(9):3683-3691.(in Chinese)
[11] 樊国燕,黄慧,李慧娟,等.中西药联用对鸡源沙门氏菌的体外抗菌活性[J].贵州农业科学,2020,48(8):73-76.FAN G Y,HUANG H,LI H J,et al.In vitro antibacterial activity of integrated application of traditional Chinese medicine and Western medicine against chicken Salmonella[J].Guizhou Agricultural Sciences,2020,48(8):73-76.(in Chinese)
[12] 昌莉丽,李华坤,刘桂兰.中西药联用对禽源大肠杆菌的体外抑菌效果研究[J].安徽农业科学,2014,42(16):5037-5038.CHANG L L,LI H K,LIU G L.Studies on the bacteriostatic effect of applying both Western and Chinese medicine on avian Escherichia coli in vitro[J].Journal of Anhui Agricultural Sciences,2014,42(16):5037-5038.(in Chinese)
[13] 孙璐,张静,尹苗,等.4种清热解毒复方中草药体外抑菌及抗PRRSV活性研究[J].中国畜牧兽医,2022,49(2):746-754.SUN L,ZHANG J,YIN M,et al.Study on the antibacterial and anti-PRRSV activities of four heat clearing and detoxifying compound Chinese herbal medicine in vitro[J].China Animal Husbandry & Veterinary Medicine,2022,49(2):746-754.(in Chinese)
[14] 王意,张博涵,陆一鸣,等.黄芩提取物对嗜水气单胞菌抑菌活性和机制的研究[J].陕西科技大学学报,2019,37(6):73-78.WANG Y,ZHANG B H,LU Y M,et al.Study on bacteriostatic activity and mechanism of Scutellaria baicalensis against Aeromonas hydrophila[J].Journal of Shaanxi University of Science，2019,37(6):73-78.(in Chinese)
[15] 张浩然,刘琴琴,徐杭忠,等.不同单味中草药与其复配组方对鲫鱼源嗜水气单胞菌的抑菌效果[J].贵州农业科学,2022,50(3):74-82.ZHANG H R,LIU Q Q,XU H Q,et al.Bacteriostatic effect of six Chinese hrbal medicines and their compounds on Aeromonas hydrophila from Carassius auratus[J].Guizhou Agricultural Sciences,2022,50(3):74-82.(in Chinese)
[16] 肖辉,苏振霞,单娟娟,等.16种中草药提取物对嗜水气单胞菌的体外抑菌实验[J].水生态学杂志,2009,30(3):53-56.XIAO H,SU Z X,SHAN J J,et al.Experimental research on anti-bacterial effect of 16 kinds of Chinese herbal medicine on Ameromonas hydrophila in vitro[J].Journal Hydtoecology,2009,30(3):53-56.(in Chinese)
[17] 田甜.鱼源嗜水气单胞菌的分离鉴定及耐药性研究[D].武汉：华中农业大学,2010.TIAN T.Isolation and identification of Aeromonas hydrophila strains from freshwater fish and their antibacterial resistance[D].Wuhan：Huazhong Agricultural University,2010.(in Chinese)
[18] 李蔼文,谭俊青,叶青,等.电镜下观察中药颗粒剂对泛耐药鲍曼不动杆菌的抑菌效果[J].按摩与康复医学,2021,12(24):63-65.LI A W,TAN J Q,YE Q,et al.The bacteriostatic effect of traditional Chinese medicine granules on pan-drug resistant Acinetobacter bowman was observed under electron microscope[J].Chinese Manipulation & Rehabilitation Medicine,2021,12(24):63-65.(in Chinese)
[19] 王唯霖.中草药提取物的抑菌机理及对大肠杆菌的耐药性研究[D].保定：河北农业大学,2015.WANG W L.Study on the antibacterial mechanism of herbal extracts and drug resistance of E.coil[D].Baoding:Hebei Agricultural University,2015.(in Chinese)
[20] 徐云凤,赵胜娟,费鹏,等.石榴皮提取物的抑菌作用及应用研究进展[J].食品与机械,2021,37(2):215-219.XU Y F,ZHAO S J,FEI P,et al.Research progress on antimicrobial effects and application of pomegranate peel extract[J].Food and Machinery,2021,37(2):215-219.(in Chinese)
[21] 刘玲丽,周紫芳,胡婕妤,等.大黄蒽醌类化合物抑菌作用研究进展[J].杭州师范大学学报(自然科学版),2021,20(1):54-58.LIU L L,ZHOU Z F,HU J Y,et al.On the antibacterial effects of anthraquinone compounds in Rhubarb[J].Journal of Hangzhou Normal University (Natural Science Edition),2021,20(1):54-58.(in Chinese)
[22] 黄铢玉,方龙香,宋超,等.抗生素恩诺沙星在渔业中的研究进展[J].农学学报,2019,9(11):57-62.HUANG Z Y,FANG L X,SONG C,et al.Antibiotic enrofloxacin in fishery:A review[J].Journal of Agriculture,2019,9(11):57-62.(in Chinese)
[23] 刘里,李林琴,汪帆.基于庆大霉素检测与分析应用的研究进展[J].化学试剂,2021,43(8):1037-1047.LIU L,LI L Q,WANG F.Recent advances on determination methods and analytical applications of gentamicin[J].Chemical Reagents,2021,43(8):1037-1047.(in Chinese)
[24] 敖蒙蒙,魏健,陈忠林,等.四环素类抗生素环境行为及其生态毒性研究进展[J].环境工程技术学报,2021,11(2):314-324.AO M M,WEI J,CHEN Z L,et al.Research progress on environmental behaviors andecotoxicity of tetracycline antibiotics[J]. Journal of Environmental Engineering Technology,2021,11(2):314-324.(in Chinese)
[25] 陈小平,王萌,杨长明,等.四环素类抗生素在我国水环境污染现状及其对水生生物的毒性研究进展[J].应用化工,2021,50(10):2780-2785.CHEN X P,WANG M,YANG C M,et al.Review of contaminations in water of tetracycline in China and toxicity on aquatic organisms[J].Applied Chemical Industry,2021,50(10):2780-2785.(in Chinese)
[26] 潘启超.头孢噻甲羧肟的药理学特性[J].中国临床药理学杂志,1989,S1:50-59.PAN Q C.The pharmacological features of ceftazidime[J].The Chinese Journal of Clinical Pharmacology，1989,S1:50-59.(in Chinese)