氟苯尼考对六种海洋致病弧菌的体外抗菌活性研究

测定了氟苯尼考等4种抗菌药对6种常见海洋致病弧菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。测试结果显示,氟苯尼考对6株弧菌的MIC值范围为2～4μg／mL,氯霉素为1～32μg／mL,土霉素为4～128μg／mL,恩诺沙星为0．125～1μg／mL;对多数弧菌氟苯尼考与氯霉素的抗菌活性相似。氟苯尼考对6株弧菌的MBC／MIC值范围为2-4,而氯霉素为2～16,表明氟苯尼考的杀菌效果优于氯霉素。对照药物恩诺沙星、土霉素的MBC／MIC值分别为1-2和2-8,恩诺沙星显示出优良的抗菌和杀菌活性。随着接种菌量增加,所测3种抗菌药的MIC随之升高;当接种量达到10^7ufu／mL时．药物对多数弧菌显示出微弱的作用。     

20种中草药对迟缓爱德华氏菌的体外抑菌试验

用琼脂扩散法和二倍稀释法测定了20种中草药对迟缓爱德华氏菌(Edwardsiella tarda)(GD080715-1、GD080715-2)的体外抗菌活性。结果表明,五倍子、乌梅、大青叶、石榴皮的抑菌作用明显,最小抑菌浓度(MIC)<6.25 mg/mL,最小杀菌浓度(MBC)<50 mg/mL;吴茱萸和菖蒲有一定的抑菌作用,MBC为50～100 mg/mL;而苦地丁、栀子的抑菌作用不明显,MBC>200 mg/mL。     

20种中草药对迟缓爱德华氏菌的体外抑菌试验

用琼脂扩散法和二倍稀释法测定了20种中草药对迟缓爱德华氏菌（Edwardsiella tarda）（GD080715-1、GD080715-2）的体外抗菌活性。结果表明,五倍子、乌梅、大青叶、石榴皮的抑菌作用明显,最小抑菌浓度（MIC）〈6.25 mg/mL,最小杀菌浓度（MBC）〈50 mg/mL;吴茱萸和菖蒲有一定的抑菌作用,MBC为50～100 mg/mL;而苦地丁、栀子的抑菌作用不明显,MBC〉200 mg/mL。     

抗菌药联用对海藻希瓦菌的体外抗菌效果

为评价硫酸庆大霉素和头孢噻肟钠对水产致病性海藻希瓦菌SFH3的体外联合抗菌效果,以6种渔用抗菌药作为对照,比较测定硫酸庆大霉素、头孢噻肟钠对海藻希瓦菌SFH3的体外抗菌作用,在此基础上进一步采用试管二倍稀释法测定分析硫酸庆大霉素和头孢噻肟钠在不同复配比例下对海藻希瓦菌SFH3的体外联合抑菌作用,并通过时间-杀菌曲线法测定其体外联合杀菌效果。试验结果显示,硫酸庆大霉素和头孢噻肟钠对海藻希瓦菌SFH3的最小抑菌质量浓度为80 mg/L和160 mg/L,抗菌活性明显优于甲砜霉素、多西环素、噁喹酸、磺胺间甲氧嘧啶钠、磺胺甲噁唑、盐酸恩诺沙星等渔用抗菌药。硫酸庆大霉素和头孢噻肟钠在复配比例8∶2和7∶3时,对海藻希瓦菌SFH3的体外联合抑菌效果表现为协同作用,对海藻希瓦菌SFH3的最小抑菌质量浓度达40 mg/L。此外,时间-杀菌曲线表明,40 mg/L硫酸庆大霉素和80 mg/L头孢噻肟钠对海藻希瓦菌SFH3的体外联合杀菌作用表现为协同效果。本研究结果证实,硫酸庆大霉素和头孢噻肟钠联用有利于提升对海藻希瓦菌SFH3的体外抗菌效果,可为研发水产致病性海藻希瓦菌的潜在控制药物提供理论参考。     

In Vitro Screening of Fungicidal Chemicals for Antifungal Activity against Saprolegnia

Saprolegnia is an important fish fungal pathogen that often results in significant economic losses to freshwater aquaculture. To find effective drugs to control saprolegniasis, 30 fungicidal chemicals used in agriculture were screened, in which kresoxim‐methyl and azoxystrobin, with minimum inhibitory concentration (MIC) values of 1.0 and 0.5 mg/L, respectively, showed good in vitro antifungal activities against Saprolegnia. Azoxystrobin has the most promising anti‐Saprolegnia activity with 50% effective concentration (EC₅₀) value of 0.212 mg/L against mycelial growth and minimum fungicidal concentration (MFC) value of 0.13 mg/L against spores, while EC₅₀ and MFC values to kresoxim‐methyl are 0.240 and 0.25 mg/L, respectively. Through the acute toxicity assay using goldfish, Carassius auratus, azoxystrobin exhibited wider margin of safety with a safe concentration (SC) value of 0.553 mg/L than kresoxim‐methyl with an SC value of 0.131 mg/L. These findings demonstrated that azoxystrobin has the potential for the development of therapy for the control of Saprolegnia in aquaculture. Both kresoxim‐methyl and azoxystrobin were tested with a post‐antifungal effects (PAFE) assay and the results revealed that the two chemicals had no significant effect on fungal growth inhibition after a 1‐hour exposure, indicating that the treatment needs to be carried out over an extended period.     

In Vitro Comparative Evaluation of Antioxidant Activity of Hydrophobic and Hydrophilic Extracts from Algicolous Fungi

The screening of potential antioxidant activities of hydrophobic (ethyl acetate) and hydrophilic (polysaccharide) extracts from both mycelia and fermentation media of eleven algicolous fungi was performed using three antioxidant assays. Algicolous fungi were able to ferment low-cost nutrients composed of potato infusion, glucose, and natural seawater and produce hydrophobic and hydrophilic metabolites with antioxidant properties. Signi?cant differences were observed in antioxidant activities of extracts from the various species evaluated using total antioxidant activity (TAA), hydroxyl radical scavenging activity (HRSA), and ferric reducing antioxidant power (FRAP), as well as their total sugar contents (TSC) of hydrophilic extracts and total phenolic contents (TPC) of hydrophobic extracts. TSC of endopolysaccharides was higher than exopolysaccharides and showed a significant correlation with both TAA and FRAP. Most of mycelial extracts were richer in phenolics than fermentation media, and TPC of hydrophobic extracts was significantly correlated with their FRAP. Mycelial extracts for most of algicolous fungi showed higher antioxidant activities than fermentation media. Both hydrophobic and hydrophilic extracts of algicolous fungi showed good antioxidant properties, especially as hydrogen-donating antioxidants, which could be considered for future applications in medicine, food production, or cosmetic industry.     

植物源杀菌剂对水产养殖常见病原菌的抑菌作用研究

植物源杀菌剂因其成分复杂、无污染、属性天然等特点,为水产养殖防治病原菌提供了新思路。本文以虎杖、千里光、黄柏、黄岑、苦参等为原材料,研制开发植物源杀菌剂,研究结果表明其对12种水产养殖常见病原菌均有较好的抑菌效果,对大肠杆菌、荧光假单胞菌的的最低抑菌浓度（MIC）为33μL／L,对金黄色葡萄球菌、痢疾志贺氏菌、温和气单胞菌的MIC为25μL／L,对肠炎沙门氏菌、单增李斯特氏菌的MIC为50μL／L,对拟态弧菌的MIC为100μL／L,对溶藻弧菌的MIC为250μL／L,对弗尼斯弧菌、副溶血弧菌的MIC为500μL／L,对创伤弧菌的MIC为1000μL／L。     

中草药对牙鲆病原迟钝爱德华氏菌的体外抑制作用研究

用倍比稀释法测定了30种中草药对牙鲆病原迟钝爱德华氏菌(LN031012-8,TH030908-5)菌株的体外抗菌活性。试验结果表明,五倍子、诃子、黄芩、秦皮和红藤的抑菌作用明显,最小抑菌浓度(MIC)<12.5 mg/ml,最小杀菌浓度(MBC)<50 mg/ml;丁香、穿心莲、虎杖、夏枯草等有一定的抑菌作用,MIC为50~100 mg/ml;而茵陈、鱼腥草、金银花、大青叶等的抑菌作用不明显,MIC均>100 mg/ml;五倍子、地榆、野菊花、马齿苋等中草药的体外抗菌活性表现了株间差异。     

黄芩对7种水产动物病原菌的体外抑菌活性研究

采用纸片扩散法和二倍稀释法,利用黄芩水提液对副溶血弧菌、嗜水气单胞菌、鳗弧菌、哈维氏弧菌、温和气单胞菌、金黄色葡萄球菌、大肠杆菌进行最低抑菌质量浓度和最低杀菌浓度的测定。试验结果表明,黄芩对鳗弧菌、嗜水气单胞菌、副溶血弧菌和金黄色葡萄球菌有较强的抑菌效果,最低抑菌质量浓度均为15.63 mg/ml,最低杀菌质量浓度分别为31.25、15.63、31.25、31.25 mg/ml;黄芩对大肠杆菌、温和气单胞菌和哈维氏弧菌的抑菌效果弱于副溶血弧菌、嗜水气单胞菌、鳗弧菌、金黄色葡萄球菌,最低抑菌质量浓度分别为31.25、62.51、31.25 mg/ml,最低杀菌质量浓度分别为62.5、62.5、125 mg/ml。     

Antibiotic Resistance of Bacterial Fish Pathogens

Antibiotic resistance by bacterial fish pathogens has been reported in all areas of aquaculture from warmwater to coldwater, and freshwater to marine environments. Decreased efficacy has been documented in many antimicrobial drugs regardless of their mechanism of action. Resistance emerges by two known genetic mechanisms, mutation on the bacterial chromosome, or extrachromosomal transfer mediated by plasmids. Alternatives to the currently used antimicrobial therapies are being evaluated for use in aquaculture, particularly the new fluoroquinolones and the third generation cephalosporins.     

5味中药醇提物对尼罗罗非鱼白细胞免疫活性的作用

将超微五倍子、白头翁、地榆、柯子、虎杖粉碎后过160目筛,按照料液比1∶10将药粉与60%的乙醇水溶液混合,再用超声波粉碎机超声1 h,用滤纸过滤收集滤液,置于多样品平行蒸发仪中浓缩后制得中药提取物。用吸取0.2 mL抗凝剂的无菌注射器自尼罗罗非鱼尾静脉采血0.5 mL,与L-15细胞培养基按体积比1∶9的比例混合成血浆,与中草药提取液单用、联用共孵育,利用氮蓝四唑还原法和流式细胞仪技术分别测定中草药提取液对尼罗罗非鱼外周血白细胞呼吸爆发活性和白细胞吞噬活性的影响。试验结果显示,五倍子、白头翁、地榆、柯子、虎杖单用均能显著提高白细胞氧呼吸爆发活性,呼吸爆发增加率分别为128.68%、123.96%、28.37%、44.44%和78.93%;试验组白细胞吞噬率分别较对照组增加44.04%、44.66%、95.00%、10.35%和64.93%。在两味中药双联用试验中,对白细胞氧呼吸爆发活性呈协同作用的组合为白头翁+地榆,呼吸爆发联用指数为1.28;对白细胞吞噬活性呈协同作用的组合为白头翁+虎杖、柯子+虎杖、五倍子+虎杖和白头翁+柯子,吞噬联用指数分别为11.45、6.20、3.03和1....     

Evaluation of In Vitro Apparent Protein Digestibility by Shrimp Using Gut Enzyme Extracts

Knowledge of apparent protein digestibility (APD) is required for optimization of feed formulae for the production of marine penaeid shrimp. The purpose of this study was to evaluate an in vitro method for determining APD in marine penaeid shrimp using gut enzyme extracts. A high correlation (r² = 0.95) was shown between single‐ingredient APD values for fish meal diets using in vivo methodology and those derived from in vitro testing of ingredients. A second study showed positive correlation (r² = 0.71) between in vitro APD of selected purified and semipurified ingredients and their reported in vivo APDs. This correlation was much higher for purified ingredients (r² = 0.93) versus less‐refined ingredients (r² = 0.24). A third trial compared in vitro APD at three different enzyme extract pH values and showed that for most protein sources, APD was significantly highest (P < 0.05) at pH = 7.0 and lower at pH = 6.1 or 7.9, indicating a neutral pH optimum for this methodology.     

盐酸恩诺沙星与其他四种抗菌药物对水产致病菌的体外抗菌作用

用琼脂平板稀释法测定了盐酸恩诺沙星、盐酸环丙沙星、氟哌酸、米诺沙星和氟苯尼考对常见水产致病菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)，结果表明：同一抗菌药物对不同致病菌的抗菌效果存在较大差异，不同抗菌药物对同一致病菌的抗菌效果也存在较大差异。使用4倍MBC的药物浓度对嗜水气单胞菌、温和气单胞菌和非O1-群霍乱弧菌杀菌时间随药物种类和致病菌不同而存在差异。     

鱼类的先天性抗菌和抗病毒机制

鱼类的皮肤是鱼类的一道物理和化学屏障，皮肤粘液除了包围入侵的微生物外，还含有许多抗菌物质，包括抗菌肽、溶菌酶、凝集素和蛋白酶。鱼类的胃肠消化道含有的酸、胆盐和酶类能使病毒和细菌灭活并消化之。多数情况下，皮肤就足以抵抗致病性微生物，而当皮肤受到机械损伤时，这些病原生物就能致病。当微生物进入鱼体组织后，会遇到一系列的可溶性的细胞成分的防御。血浆里的补体系统在识别细菌的过程中起核心作用。其激活产物能裂解细菌，激发炎症，引起吞噬细胞的浸润，提高其吞噬活力。补体能被细菌的组分或其代谢产物直接激活，也可以被C-反应蛋白，凝集素等能直接结合细菌表面的物质间接激活。血浆也含有许多能抑制细菌生长的物质（例如转铁蛋白，抗蛋白酶）或杀菌物质（例如溶菌酶）。鱼类感染病毒后，感染的细胞产生干扰素，引起相邻的细胞的抗病毒机制以免受感染。抗病毒的细胞毒性细胞能裂解病毒感染细胞，降低病毒在其中增殖的速率。因此，先天性防御可以提供一种预先存在的，快速反应的保护力，该系统是非特异性的；与反应缓慢的、温度依赖性的特异性免疫相比，具有很多优势。     

地锦草对水产病原菌体外抗活性的研究

用95%的乙醇来提取地锦草(Euphorbia humifusa Wild)中的化合物,再分别用石油醚、乙酸乙酯、正丁醇和水对地锦草乙醇提取物进行分极性段萃取,研究地锦草各极性段二甲基亚砜萃取溶液对嗜水气单胞菌(Aeromonas hydrophila)、温和气单胞菌(Aeromonas sobria)、柱状屈挠杆菌(Flexibacter calumnaris)、鳗弧菌(Vibrisis angnillarumz)4种常见水产动物病原菌的抑菌效果。结果表明,地锦草各极性段的抑菌作用强度有明显差别(P<0.05),其中乙酸乙酯、石油醚极性段的抑菌作用最为强烈,对温和气单胞菌的MIC均≥12.5mg/mL;乙酸乙酯极性段对嗜水气单胞菌、鳗弧菌的MIC均≥50mg/mL;石油醚极性段对嗜水气单胞菌、鳗弧菌的MIC≥25mg/mL;正丁醇和水极性段对4种菌的MIC均≥50mg/mL。     

黄姜杀灭鱼类指环虫活性部位的初步研究

以寄生于金鱼鳃部的指环虫为指示寄生虫,采用活体感染、活体杀虫的方法,通过不同极性的溶剂回流提取黄姜,制备粗提物,进行杀灭金鱼指环虫的药效活性追踪试验,确定黄姜杀灭鱼类指环虫的活性部位,并对活性部位进行安全性评价.试验结果表明,黄姜的杀虫活性部位是70%乙醇部位,其质量浓度为20.0 mg/L时,平均最高杀虫率为100%.70%乙醇提取物经过有机溶剂进一步萃取后,杀虫药效试验表明,石油醚和最终水相萃取部位对指环虫具有杀灭作用,但石油醚萃取部位作用较佳,其质量浓度为5.0 mg/L时,平均最高杀虫率为100%.活性部位对草鱼急性毒性试验结果显示,石油醚萃取部位对草鱼的48 h半致死质量浓度(LC50)为33.54 mg/L,其安全质量浓度为9.64 mg/L,表明黄姜是一种比较安全,杀虫效果良好的水产用中草药.     

洛河水域光合细菌的分离鉴定及体外抑菌作用研究

光合细菌作为重要的微生物资源,对它进行研究开发应用具有重要的意义。本试验从光合细菌的分离鉴定及抑菌作用出发,以期得到较好的菌株用于水产养殖中。试验通过选择性富集培养的方式从洛阳市洛河分离得到光合细菌,通过形态学观察,以伯杰氏细菌鉴定手册为依据进行鉴定,鉴定结果为：污水中培养物为沼泽红假单胞菌,清水中培养物为万尼氏红微菌。体外抑菌试验表明分离得到的光合细菌对G+ 、G- 模式菌种大肠杆菌和金黄色葡萄球菌都有较强的抑菌活性。     

几种海藻多酚对鱼类致病菌的抑菌性研究

用平板生长抑制法对角叉菜、海蒿子及鼠尾藻等藻体中的多酚化合物进行抑菌性试验,观察其对海神弧菌、巴西弧菌、徐氏弧菌、查格斯氏弧菌、鳗弧菌、波氏弧菌及哈维氏弧菌的抑菌效果。试验结果表明,不同来源海藻多酚对受试海洋弧菌的抑制能力具有显著的差异(P0.05);多酚含量越高,抑菌能力越强;多酚含量和多酚来源具有显著的交互效应(P0.05),随着多酚含量的增加,抑制差异显著增强;同一来源多酚对不同受试海洋弧菌的抑制差异性不显著(P0.05)。     

中草药体外抑杀大鲵致病性豚鼠气单胞菌的药效研究

筛选对大鲵致病性豚鼠气单胞菌有抑菌作用的中草药。采用琼脂打孔法,测定中草药在不同pH值时对大鲵致病性豚鼠气单胞菌的体外抑菌圈效果。使用二倍稀释法,测定中草药在不同pH值时对大鲵致病性豚鼠气单胞菌的最小抑菌质量浓度和最小杀菌质量浓度。试验结果表明,200种中草药对大鲵致病性豚鼠气单胞菌的抑菌效果各不相同,其中135种中草药具有一定抑菌作用,其余65种无抑菌作用,而不同pH值时抑菌效果亦有所不同。药液为酸性时,乌梅、五味子、五倍子及诃子的抑菌效果极强,抑菌圈明显。135种中草药对豚鼠气单胞菌的最小抑菌质量浓度和最小杀菌质量浓度亦有所差异,其中,诃子、五倍子、石榴皮、大黄及槐角的最小抑菌质量浓度小于1mg/mL,抑菌作用最强。本试验筛选出具有很强抑菌作用的中草药。     

条斑紫菜蛋白酶解多肽的抑菌活性

以条斑紫菜为原料,分别提取水溶、酸溶、碱溶、盐溶性蛋白质,并用6种蛋白酶酶解,将所得24种酶解物采用琼脂孔穴扩散法对金黄色葡萄球菌(Staphylococcus aureus)、四联微球菌(Micrococcus tetragenus)、枯草杆菌(Bacillus subtilis)以及大肠杆菌(Escherichia coli)4种菌进行抑菌活性测定。结果显示,水溶性胃蛋白酶酶解物的抑菌活性最优。将水溶性蛋白质经硫酸铵沉淀以及超滤分级,比较不同饱和度硫酸铵沉淀所得蛋白质酶解物以及不同分子量多肽的抑菌效果。结果显示,水溶性蛋白在硫酸铵饱和度为40%、50%时沉淀所得蛋白的胃蛋白酶酶解物中,相对分子质量小于5 k Da的级分抑菌效果最为明显。此外,对该级分进行了抑菌活性影响因素的初步研究,结果表明该级分热稳定性以及酸碱稳定性均较好,有望开发为天然的食品防腐剂。