广东五种常见中草药的体外抑菌试验

为测定鸭脚木皮、木蝴蝶、火炭母、岗梅根、倒扣草（鲜）这5种广东常见中草药的体外抑菌活性,采用琼脂平板打孔法、试管倍比稀释法和平板培养法,分别测定其对金黄葡萄球菌、大肠杆菌、沙门氏菌、链球菌、枯草芽孢杆菌和蜡状芽孢杆菌的抑制效果.结果表明：大肠杆菌、链球菌对火炭母呈高度敏感,最小抑菌浓度（MIC）分别为1/40、1/80,最小杀菌浓度（MBC）分别为1/40、1/80;链球菌对鸭脚木皮、木蝴蝶均呈高度敏感,最小抑菌浓度（MIC）分别为1/20、1/10,最小杀菌浓度（MBC）分别为1/20、1/10;岗梅根、倒扣草（鲜）对六种菌均无抑制作用.     

不同植物精油体外抑菌效果的研究

植物精油是一种天然绿色活性物质,具有很好的抑菌效果,其在饲料中应用具有广阔开发前景。本文采用琼脂平板孔洞法分别对不同来源的肉桂醛、香芹酚等植物精油及其混合物进行体外抑菌效果筛选和评估。结果表明,香芹酚、肉桂醛、百里香酚和丁香酚对大肠杆菌、沙门氏菌、金黄色葡萄球菌等常见动物消化道内有害菌有很好的抑菌效果,其抑菌顺序为肉桂醛>香芹酚>丁香酚>百里香酚,用棕榈油进行1:4比例稀释后,精油的抑菌效果均有不同程度的下降。由4种精油组成的复方产品对大肠杆菌等肠道有害菌有较好的抑制作用,且热稳定性好。结论:肉桂等植物精油对常见有害菌具有良好的抑菌效果,可作为天然的抑菌剂。     

酢浆草等11种中草药的体外抑菌试验

试验选用酢浆草、乌梅、桂枝、板蓝根、黄芩等11种中药,采用常规中药药敏试验打孔法和试管法对大肠杆菌、金黄色葡萄球菌、枯草杆菌、链球菌、肠炎沙门氏菌等7种细菌进行体外抑菌试验.结果显示乌梅、秦皮、桂枝、大黄对上述7种菌都有较强抑菌效果.酢浆草除对链球菌无抑菌效果外,对其它细菌均呈高度敏感.黄芩对肠炎沙门氏菌呈高度敏感.其它药物对上述7种菌抑菌效果较差或无抑制作用.     

常用中草药对水产动物致病菌体外抑菌效果研究

试验旨在探讨黄柏、诃子、知母、苏木等常用中草药对水产动物致病菌的体外抑菌效果,为今后水产养殖细菌性疾病的防控提供参考。以美人鱼发光杆菌、嗜水气单胞菌、哈维氏弧菌、创伤弧菌临床分离株为试验菌株,以抑菌圈直径、最小抑菌浓度（MIC）、最小杀菌浓度（MBC）及联合抑菌指数（FICI）作为检测指标,研究常用中草药对水产病原菌的抑菌活性。采用水提法提取中草药有效成分,并通过琼脂平板打孔法测定中草药对水产致病菌抑菌圈,微量肉汤稀释法测定中草药MIC和MBC及微量肉汤棋盘稀释法测定联合抑菌指数（FICI）。结果显示,16种中草药对水产致病菌具有不同程度的抑菌作用。药敏试验结果显示,苏木、女贞子、夏枯草、青皮对4种水产致病菌均具有抑菌效果,抑菌圈直径在10.5~29.0 mm之间,而4种病原菌对马齿苋均表现为耐药;MIC检测结果显示,苏木、女贞子、夏枯草、青皮对4种病原菌的MIC值为7.81~250.00 mg/mL;MBC检测结果显示,苏木、女贞子、夏枯草、青皮对4种病原菌的MBC值为7.81~500.00 mg/mL;联合抑菌试验结果显示,苏木与夏枯草、女贞子及青皮联用仅对美人鱼发光杆菌起到协同抑菌作用,对嗜水气单胞菌、创伤弧菌及哈维氏弧菌均无抑菌作用,女贞子和青皮组合仅对哈维氏弧菌抑菌作用为相加作用,该组合在抑菌效果上对其他病原菌表现为无关或颉颃作用,其他药物组合在抑菌效果上对4种病原菌则表现为无关或颉颃作用;联合抑菌圈测定结果表明,中草药联合使用对4种病原菌抑菌作用不一。本试验结果表明常用中草药对水产动物致病菌具有一定程度的抑菌作用。     

溪黄草与桉树叶的体外抑菌试验

本试验应用琼脂平板打洞法、试管稀释法和平板培养法测定溪黄草和桉树叶煎剂对大肠杆菌、金黄色葡萄球菌、蜡状芽孢杆菌、枯草芽孢杆菌、链球菌等几种常见病原菌的体外抑制作用。结果显示,溪黄草煎剂对大肠杆菌、金黄色葡萄球菌、蜡状芽孢杆菌、枯草芽孢杆菌、链球菌均显示高度敏感,最小抑菌浓度(MIC)分别为1/80、1/320、1/160、1/80、1/80,最小杀菌浓度(MBC)分别为1/80、1/80、1/160、1/40、1/80;桉树叶煎剂对大肠杆菌、金黄色葡萄球菌、蜡状芽孢杆菌、枯草芽孢杆菌、链球菌均显示高度敏感,最小抑菌浓度(MIC)分别为1/80、1/80、1/40、1/160、1/40,最小杀菌浓度(MBC)分别为1/80、1/80、1/10、1/40、1/20。结果表明,溪黄草和桉树叶煎剂对常见的多种细菌均有较强的抑菌作用。     

迷迭香酸与抗菌药联合对含fosA3基因细菌抑菌效果的研究

试验旨在探讨迷迭香酸与抗菌药联合体外抗含fosA3耐药基因大肠杆菌的效果。本试验对临床分离的细菌进行鉴定,确定细菌所含耐药基因种类,用牛津杯法测定迷迭香酸的抑菌活性,平板涂布法测定迷迭香酸的杀菌活性,采用96孔反应板二倍微量稀释法分别检测迷迭香酸与抗菌药的最小抑菌浓度(MIC),再用微量棋盘稀释法测定迷迭香酸与抗菌药联合作用后的分级抑菌浓度指数(FICI)。结果显示,分离到含fosA3耐药基因大肠杆菌;迷迭香酸的MIC为640 μg/mL,迷迭香酸与环丙沙星、左旋氧氟沙星、黏杆菌素、加替沙星、阿米卡星、头孢他啶、头孢噻呋钠联合应用后其FICI≤0.5,有协同作用;与头孢曲松钠、诺氟沙星、痢菌净联合应用后0.5fosA3基因大肠杆菌有一定的抑菌活性和杀菌活性,其与部分抗菌药联合应用后,抗菌药体外抗菌活性显著增强。     

植物精油与丁酸钠的体外协同抑菌效果研究

本试验旨在研究植物精油与丁酸钠的体外协同抑菌效果。采用牛津杯法,测定肉桂醛、百里香酚和香兰素对于4种细菌(大肠杆菌、沙门氏菌、产气荚膜梭菌、乳酸菌)的抑菌效果。并将单方和复合植物精油分别与丁酸钠复配,测定其抑菌效果。结果表明:3种植物精油均对大肠杆菌、沙门氏菌、产气荚膜梭菌有明显抑制作用,但对乳酸菌无抑制作用,在相同浓度下,抑菌效果为肉桂醛香兰素百里香酚。肉桂醛与香兰素复合植物精油抑菌效果优于单方植物精油,当肉桂醛与香兰素的质量比为6∶1时抑菌效果最佳。丁酸钠没有显示出抑菌作用,但丁酸钠与植物精油复合可明显降低植物精油的使用量。综上所述,不同植物精油的复合具有协同作用,植物精油和丁酸钠之间也存在着协同作用。     

中草药对肠炎沙门氏菌的体外抑菌试验

用定量分析的方法探讨单味及多味复方中草药的药理活性,为临床应用提供理论依据.用2倍稀释法测定13种中草药对肠炎沙门氏菌的体外MIC;将13种中药两两组合后测定MIC,依据结果组成8个复方,测定它们及5个中药方剂的MIC.结果单味中草药对肠炎沙门氏菌的MIC在15.6～500 g/L之间;78种两味中药组合的MIC在15.6～500 g/L之间,少数组合有拮抗作用;13种3～10味中药组合MIC在15.6～250 g/L之间,大部分组合表现协同作用,显示出中药配伍优势.     

细菌敏感精油的筛选及与乳化剂组合的抑菌效果研究

为了筛选常见病原菌敏感的精油和乳化剂,首先采用牛津杯法测定了12种精油对产气荚膜梭菌、大肠埃希氏菌和鼠伤寒沙门氏菌的抑菌作用,然后将筛选出的5种抑菌效果较好的精油与吐温系列乳化剂分别按不同比例互作,测定对上述3种细菌的最小抑菌浓度(MIC),最后根据MIC结果,测定产气荚膜梭菌在低于精油MIC下的生长曲线以及在MIC下的菌体形态变化。结果显示,肉桂醛对产气荚膜梭菌最敏感,且10%吐温40乳化肉桂醛可显著降低对产气荚膜梭菌的MIC;生长曲线表明1/2、1/4、1/8MIC的肉桂醛对产气荚膜梭菌的生长均有明显抑制作用;扫描电镜显示肉桂醛作用后的产气荚膜梭菌形态发生明显改变。肉桂醛对产气荚膜梭菌的抑菌效果优于其他精油。试验结果为临床防治产气荚膜梭菌引发的梭菌性肠炎提供了科学依据。     

In vitro susceptibility of Brachyspira hyodysenteriae to organic acids and essential oil components

The antibacterial potential of organic acids and essential oil components against Brachyspira hyodysenteriae, the causative pathogen of swine dysentery, was evaluated. Minimum inhibitory concentrations (MIC) of 15 compounds were determined at pH 7.2 and pH 6.0, using a broth microdilution assay. In addition, possible synergism was determined. MIC values for the three tested strains were similar. For organic acids, MIC values at pH 6.0 were lower than at pH 7.2. B. hyodysenteriae was most sensitive to cinnamaldehyde and lauric acid, with MIC values <1.5 mM. Most antibacterial effects of binary combinations were additive, however, for thymol and carvacrol, synergism could be observed. In vitro results demonstrate the antibacterial action of certain essential oil components and organic acids against B. hyodysenteriae.     

Rosemary oil,clove oil,and a mix of thyme-orange essential oils inhibit Salmonella and Campylobacter in vitro

The demand for foods that are free of pathogens and chemical residues has increased interest in the use of plant-based products as natural antimicrobials. Essential oils (EO) from plants are natural compounds that have been shown to have antimicrobial properties against food-borne pathogens. The objective of the current study was to determine the ability of various concentrations of 4 selected EO to inhibit Salmonella enterica (3 different serovars and a cocktail of all 3) and Campylobacter (2 strains of Campylobacter jejuni, one strain of Campylobacter coli, and a cocktail of all 3). The disc diffusion method was used to screen the oils of thyme, orange, rosemary, and clove oil. The minimum inhibitory concentration or minimum bactericidal concentration of the EO was determined using a 2-fold broth dilution method at concentrations ranging from 0.0008 to 1.000% (vol/vol). Two independent experiments were performed. Zones of inhibition (ZI) were expressed in millimeters and concentrations were expressed in percentages. All the oils demonstrated antibacterial activity against the strains tested. However, thyme oil demonstrated the strongest inhibitory activity than other oils against Salmonella (ZI of 18.5 mm). In general, Campylobacter was more susceptible to the antibacterial activity of EO, with plates containing thyme or clove oil showing no growth. Orange oil was also highly effective on Campylobacter, with a mean ZI of 17.5 mm. The least expensive treatment effective against both Salmonella and Campylobacter was a combination of 100% concentrations of thyme and orange oil combined on a 50:50 proportion. Tested on the same strains of bacteria, the thyme-orange combination (TOC) had a mean ZI of 20.5 mm for Salmonella and 21.3 mm for Campylobacter. Thyme-orange combination demonstrated a synergetic effect against Salmonella, but no such effect was noticed for Campylobacter. On average, 0.14% TOC was required to inhibit both pathogens. Hence, TOC can be considered as a potential antimicrobial for future studies on food systems.     

纳米材料抗菌性能的研究

试验以硒化铜粉体作为抗菌剂,选用大肠杆菌和金黄色葡萄球菌作为试验菌种,采用抑菌圈法、最低抑菌浓度法、最低杀菌浓度法对纳米材料的抗菌性能进行了定性和定量研究。抑菌圈试验结果表明,硒化铜具有良好的抗菌性能。最低抑菌浓度试验结果表明,硒化铜对大肠杆菌和金黄色葡萄球菌的最低抑菌浓度均为30 mg/L。最低杀菌浓度结果表明,硒化铜对大肠杆菌和金黄色葡萄球菌的最低杀菌浓度分别为220,180 mg/L,当达到该浓度时灭菌率均可达到99%以上。     

15种植物精油添加剂热稳定性和体外抑菌作用研究

肉桂醛、香芹酚、丁香酚、百里香酚等植物精油活性物质具有广谱抑菌作用,在兽用抗菌药物减量化使用试点行动推动下,以植物精油为核心的添加剂产品越来越多。本研究旨在比较市售的15种猪、禽用植物精油添加剂的热稳定性,并探究它们对畜禽生产中常见病原菌的体外抑菌作用,以期为植物精油添加剂的合理使用提供参考。分别以多重耐药的猪源沙门菌、大肠杆菌和链球菌以及禽源沙门菌和大肠杆菌为研究对象,以大肠杆菌ATCC25922为质控菌,采用微量肉汤法体外测定肉桂醛、香芹酚、丁香酚、百里香酚和各精油添加剂的最小抑菌浓度(MIC)。通过棋盘稀释法比较肉桂醛、香芹酚、丁香酚和百里香酚不同复配组合对猪源大肠杆菌的抑菌效果。将每种添加剂等量分为对照组和热处理组,分别经乙醚抽提和气相色谱仪分析后,对每种添加剂中肉桂醛、香芹酚、丁香酚和百里香酚及其总量进行测定比较。结果显示:6株供试菌株对肉桂醛、香芹酚、丁香酚和百里香酚均表现出高度敏感性(MIC值0.25～2 mg/mL);当香芹酚:百里香酚和丁香酚?百里香酚复配比例为2∶1时表现出协同作用(FICI=0.5);植物精油添加剂EG2对6株供试菌无明显抑菌作用,其余14种产品的MIC值为1～32 mg/mL。此外,15种添加剂含肉桂醛、香芹酚、丁香酚和百里香酚的总量范围在2.64～691.40 mg/g;15种产品热处理前后精油总量变化范围在0.2%～14.11%。综上所述,受试精油添加剂产品普遍热稳定性较好,并具有一定的抑菌作用,且抑菌效果与产品肉桂醛、香芹酚、丁香酚、百里香酚和精油总量存在一定的正相关;EG7、EG3、ES2、ES1、EAP2和EP1这6种产品的体外抑菌效果明显,有潜力用于预防和治疗多重耐药菌引起的猪禽疾病。     

Study on the Effect of the Rosemarinic Acid Combined with Antibacterial Agents on the Bacterial with Gene fosA3

This research was aimed to investigate the effect of rosemarinic acid combined with antibacterial agents against bacterial carrying gene fosA3 in vitro.Resistance gene types were determined by identification of bacteria isolated in clinics.Antibacterial activity and fungicidal activity of rosemarinic acid were tested by oxford cup method and spread-plate method.The minimal inhibiton concertration (MIC) of rosemarinic acid and antibacterial agents were tested through twice micro-dilution method,fractional inhibitory concentration index (FICI) of rosemarinic acid combined with antibacterial agents were determined by microdilution checkerboard techniques.Escheriohia coli carrying multidrug resistance gene fosA3 was isolated.The MIC of rosemarinic acid was 640 μg/mL,when application of rosemarinic acid combined with ciprofloxacin,levofloxacin,coly-mycin,gatifloxacin,amikacin,ceftazidime,ceftiofur sodium,FICI≤0.5,showed an additive effect;With ceftriaxone sodium,norfloxacin,mequindox,0.5fosA3 Escheriohia coli.Antibacterial activities of antibacterial agents against the fosfomycin resistance gene fosA3 Escherichia coli were enhanced significantly by rosemarinic acid.     

阿散酸-TMP的制备与体外抗菌活性研究

以阿散酸和甲氧苄啶(TMP)为原料,制备阿散酸-TMP,采用IR、熔点对其进行了表征;正交试验确定了最优反应条件为:反应温度80 ℃,反应时间3 h,反应物料比1.1∶1(阿散酸∶TMP)。通过体外抑菌试验来测试其抑菌效果,结果表明,阿散酸-TMP对5种供试菌的抑菌圈直径都在20 mm以上,显著大于阿散酸与TMP的混合物(P＜0.05);阿散酸-TMP对埃希氏大肠杆菌、金黄色葡萄球菌、鸡白痢沙门氏菌、福氏志贺氏菌、藤黄微球菌的MIC分别为32、64、256、32、128 μg/mL,均小于阿散酸与TMP的混合物。阿散酸-TMP对供试菌的抑菌效果好于阿散酸、TMP及阿散酸与TMP的混合物。     

Comparing the minimum inhibitory and mutant prevention concentrations of selected antibiotics against animal isolates of Pasteurella multocida and Salmonella typhimurium

Historically, the use of antibiotics was not well regulated in veterinary medicine. The emergence of antibiotic resistance (ABR) in pathogenic bacteria in human and veterinary medicine has driven the need for greater antibiotic stewardship. The preservation of certain antibiotic classes for use exclusively in humans, especially in cases of multidrug resistance, has highlighted the need for veterinarians to reduce its use and redefine dosage regimens of antibiotics to ensure efficacy and guard against the development of ABR pathogens. The minimum inhibitory concentration (MIC), the lowest concentration of an antibiotic drug that will prevent the growth of a bacterium, is recognised as a method to assist in antibiotic dosage determination. Minimum inhibitory concentrations sometimes fail to deal with first-step mutants in bacterial populations; therefore dosing regimens based solely on MIC can lead to the development of ABR. The mutant prevention concentration (MPC) is the minimum inhibitory antibiotic concentration of the most resistant first-step mutant. Mutant prevention concentration determination as a complementary and sometimes preferable alternative to MIC determination for veterinarians when managing bacterial pathogens. The results of this study focused on livestock pathogens and antibiotics used to treat them, which had a MIC value of 0.25 µg/mL for enrofloxacin against all 27 isolates of Salmonella typhimurium. The MPC values were 0.50 µg/mL, with the exception of five isolates that had MPC values of 4.00 µg/mL. The MPC test yielded 65.52% (18 isolates) Salmonella isolates with florfenicol MICs in the sensitive range, while 11 isolates were in the resistant range. Seventeen isolates (58.62%) of Pasteurella multocida had MIC values in the susceptible range and 41.38% (12 isolates) had an intermediate MIC value. Mutant prevention concentration determinations as done in this study is effective for the antibiotic treatment of bacterial infections and minimising the development of resistance. The MPC method can be used to better control to prevent the development of antibiotic drug resistance used in animals.     

4味中药及其与抗菌药的复方制剂的MIC测定

用改进的试管两倍稀释法测定了黄连、鱼腥草、大青叶和苦参4味中药及其相互配伍（1：1）对标准大肠杆菌的最小抑菌浓度和4味中药与抗菌药配伍（1000：1）对临床分离鸡大肠杆菌的最小抑菌浓度。结果表明，4味中药及其相互配伍对大肠杆菌标准株有一定的抑制作用，其MIC值介于3．91—62．50mg／mL之间，其中大青叶和鱼腥草以1：l配伍对大肠杆菌抑菌效果最好，MIC为3．91mg／mL；对临床分离株，单方药中黄连抑菌效果最好，MIC为31．25mg／mL，24种复方药中，黄连和盐酸多西环素或加替沙星以1000：1配伍的抑菌效果最好，其MIC为3．91mg／mL。