亚抑菌浓度氟苯尼考作用下质粒pSD11对大肠杆菌适应性的影响

通过分析cfr阳性质粒pSD11对大肠杆菌的适应性影响,并测定氟苯尼考对携带质粒pSD11的大肠杆菌MG1655(Escherichia col MG1655)的最低筛选浓度(MSC),评价亚抑菌浓度氟苯尼考作用下多重耐药基因cfr在大肠杆菌中的持续存在和扩散传播的风险.通过电转化的方法将质粒pSD11导入模式菌E.coli MG1655中,获得耐药菌株E.coli MG1655/pSD11.通过测定生长曲线,比较无抗生素作用下耐药菌株E.coli MG1655/pSD11与同源敏感菌株E.coli MG1655的生长动力学参数,评价质粒pSD11对宿主菌适应性的影响.并通过比较分析不同亚抑菌浓度(0.031 25,0.062 5,0.125,0.25,0.5,1.0和2.0μg/mL)氟苯尼考作用下的生长速率,测定氟苯尼考对携带质粒pSD11的宿主菌最小筛选浓度(MSC).携带质粒pSD11的耐药菌E.coli MG1655/pSD11的延滞时间(2.853 3±0.031 8 h/λ)相对于同源敏感菌E.coli MG1655极显著延长(2.825 3±0.024 4 h/λ),且世代时间也显著延长(分别为:1.705 7±0.006 4 h/G和1.526 9±0.007 8 h/G).氟苯尼考对耐药菌E.coli MG1655/pSD11的MSC为0.042 5μg/mL约为相应敏感菌的MICsus的1/100.本研究表明携带cfr基因的质粒pSD11在无抗生素选择下对宿主菌有明显的适应性影响,但是在极低的氟苯尼考浓度的作用下存在选择优势,能够平衡适应性成本使宿主菌得到富集与传播,以期为对该质粒在环境中的传播风险评估及氟苯尼考的临床应用提供的参考.(图2表2参18)     

蚯蚓抗菌肽EABP-1的分离纯化及部分性质

经硫酸铵沉淀、超滤和阳离子交换分离 ,得到了一蚯蚓抗菌肽EABP 1,该肽的最大紫外吸收在 2 77.16nm ,SDS PAGE结果表明其Mr≈ 2 0× 10 3,精确的分子量没有确定 .最小抑菌浓度 (ρMIC)实验表明 ,EABP 1对鹑鸡肠球菌(Enterococcusgallinarum)、绿脓杆菌 (Pseudomonaspyocyanea)、鲍氏不动 (Acinetobacterbaumanii)、土生克雷伯 (Kleb siellaterrigena)的 ρMIC为 11.4μg/mL ,对粪肠球菌 (Enterococcusfaecalis)的 ρMIC为 2 2 .8μg/mL ,对真菌白色念株菌(Candidaalbicans)没有表现为完全的抑制作用 .图 1表 1参 10     

链格孢菌毒素细交链孢菌酮酸对蚕豆根尖细胞微核和有丝分裂的影响

利用蚕豆根尖为材料,研究了链格孢菌毒素细交链孢菌酮酸对细胞微核和有丝分裂的影响,以评价其细胞遗传毒性.结果表明,当浓度为12.5～50μg/mL时,细交链孢菌酮酸对蚕豆根尖细胞微核率和有丝分裂指数不产生显著影响,微核率低于3‰,有丝分裂指数超过9.5%.浓度升高到100～400μg/mL时,细交链孢菌酮酸使蚕豆根尖细胞微核率显著升高,有丝分裂指数显著下降,其中,400μg/mL时,微核率高达36.01‰,有丝分裂指数仅为1.90%.当细交链孢菌酮酸浓度为12.5～100μg/mL时,与对照相比,蚕豆根尖细胞多核仁率无显著差异,多核仁率低于3‰;浓度升高到200～400μg/mL时,细胞多核仁率显著升高,400μg/mL时,多核仁率高达26.10‰.     

光能变价离子钛对多种细菌的抑制作用

光能变价离子钛(Titanium ion of variable valence with light energy,TIVL)对环境友好、对人畜无害,能有效减少作物的发病率及发病症状.为了解其抑菌活性,采用不同浓度的TIVL对5种细菌,大肠杆菌(Escherichia coli)、娄彻氏链霉菌(Streptomyces rochei)、根癌农杆菌(Agrobacterium tumefaciens)、成团泛菌(Pantoe aagglomerans)和茄科雷尔氏菌(Ralstonia solanacearum),进行平板抑菌试验研究.结果显示,在一定菌液浓度(2×10~3-10~4 CFU/mL)下,16mg/L的TIVL即可显著抑制这5种菌绝大部分菌落的定植和生长.TIVL浓度为4 mg/L时,即可有效抑制娄彻氏链霉菌,抑菌率达97.33%;浓度为8 mg/L时,99.15%大肠杆菌和89.16%根癌农杆菌被抑制;浓度为16 mg/L时,87.68%成团泛菌和74.06%茄科雷尔氏菌被抑制.结果表明TIVL在低浓度下即对上述5种菌有良好的抑制作用,推测TIVL具有广谱抑菌活性;本研究可为进一步研究TIVL抑菌机理及农业生产应用提供依据,为农药减施、绿色农作物产品生产提供新途径.     

中药三黄汤、小檗碱对E.coli生长抑制作用与庆大霉素的比较

庆大霉素对E．coli的抑菌作用强，试管稀释法、琼脂稀释法和平板抑菌圈法抑菌试验得到一致的MIC(3-5μg／mL)；三黄汤、小檗碱抑菌作用弱而稳定，试管稀释法测得的MIC分别为500mg／mL和2．5mg/mL，而用平板抑菌圈法得不到明显的抑菌圈．在亚MIC药物液体分批培养时，庆大霉素抑菌曲线起初2h迅速下降，而后回升到初始浓度，而三黄汤抑菌曲线平缓稳定，说明中药和抗生素的抑菌机制明显不同．庆大霉素与小檗碱混合使用时，与庆大霉素单独使用相近，而与三黄混合使用时，则主要表现三黄汤的抑菌作用．因此中西药结合时需要慎用．在接近MIC药物培养基中连续传代20次后，庆大霉素MIC提高8倍多，而三黄汤和小檗碱MIC无显著变化，无抗药性产生，也不产生对庆大霉素的交叉抗药性．研究结果还表明，在1／4MIC三黄汤中连续传代20次，能消除E.coli已形成的庆大霉素抗性，这为解决抗生素抗药性提供了线索．图4参23     

柠檬醛体外抗MRSA活性及其对β-内酰胺类抗生素的增效作用

通过体外抑菌实验评价柠檬醛单独使用以及与3种β-内酰胺类抗生素(阿莫西林、头孢氨苄及头孢吡肟)联合使用对耐甲氧西林金黄色葡萄球菌(Methicillin-resistant Staphylococcus aureus,MRSA)的抑制效果.采用微量稀释法测定柠檬醛与3种β-内酰胺类抗生素的最低抑菌浓度(Minimum inhibitory concentration,MIC)及最低杀菌浓度(Minimum bactericidal concentration,MBC);用微量棋盘法测定柠檬醛与3种β-内酰胺类抗生素的分级抑菌浓度(Fractional inhibitory concentration,FIC)指数;用分光光度法测定柠檬醛和阿莫西林联合使用后对MRSA生长的抑制效果.结果显示:柠檬醛MIC和MBC分别为695-2 780μg/mL和1 390-2 780μg/mL.联合药敏试验发现柠檬醛与3种β-内酰胺类抗生素联合使用FIC指数集中分布在≤0.5和0.5-1;与单独用药相比,阿莫西林、头孢氨苄和头孢吡肟的MIC_(50)分别降低为原来的1/9、1/15和1/9;1/4 MIC阿莫西林与1/4 MIC柠檬醛联合用药后,显著抑制了MRSA的生长.因此,柠檬醛具有一定的体外抗MRSA活性,与阿莫西林、头孢氨苄和头孢吡肟等β-内酰胺类抗生素联合使用时可增强β-内酰胺类抗生素的活性,降低其用量.     

4种中药单体对IncX4型质粒pSD11稳定性的影响

以携带多重耐药基因cfr的IncX4型质粒pSD11为试验材料，研究小檗碱、黄芩苷、芦荟大黄素、姜黄素4种中药单体对耐药质粒稳定性的影响，为中药单体消除病原菌耐药性提供理论依据.通过测定最小抑菌浓度（MIC）及药物作用下携带质粒pSD11的大肠杆菌的生长曲线和生长动力学参数，分析中药单体的抑菌效果.结合实时荧光定量PCR(RTPCR)及平板计数研究药物对质粒稳定性的影响，同时检测胞外乳酸脱氢酶和碱性磷酸酶的含量分析中药单体对细胞壁、细胞膜的影响.结果显示，小檗碱、黄芩苷、芦荟大黄素对大肠杆菌MG1655/pSD11(Escherichia coli MG1655/pSD11)的MIC均为1 280μg/mL，姜黄素> 160μg/mL.4种中药单体对E. coli MG1655/pSD11具有一定的抑菌效果，生长动力学参数结果表明1/2 MIC小檗碱和黄芩苷显著延长E. coli MG1655/pSD11的滞后期；1/2 MIC姜黄素和1/2、1/4MIC芦荟大黄素极显著降低E. coli MG1655/pSD11的最大生长速率、延长时代时间.经1/2和1/4 MIC的小檗碱和黄...     

植物挥发性物质成分分析及抑菌作用研究

对植物挥发性物质的化学成分及其抵抗微生物的能力以及与生态学相关可能产生的增效作用进行了研究。通过采用活体采气法收集20种园林植物的挥发性物质,利用气质联用仪测定物质成分结果,再进一步做抑菌作用研究。根据测定结果,实验最终确定的抑菌化学成分有:间羟基苯甲酸(m-hydroxybenzoic acid),水杨酸甲酯(methyl salicylate),邻苯二酚(catechol),乙酸(acetic acid),乙醚(acetaldehyde),丙烯酸(acrylic acid),乙硫醚(diethyl sulphide),莰酮-〔2〕(camphanone-2),莰醇-〔2〕(borneol-2)。同时,采用纸片扩散法和MIC(最小抑菌浓度)法对药品进行抑菌实验。结果表明:莰酮-〔2〕(樟脑)的抑菌效果最显著也最全面,水杨酸甲酯的抑菌效果最不显著也最不全面。丙烯酸、邻苯二酚、莰酮—[2]的最佳抑制细菌是金黄色葡萄球菌和酵母菌,质量浓度为0.156×10-3 mg.L-1时即可有效抑制这些微生物的繁殖,可根据这一特点,在医疗、食品中加以运用。从环境生态学角度看,在某些以空气传播的流行性疾病的多发地,特别是在呼吸道疾病曼延的地区,多栽种一些抑菌作用较强的植物(如银杏、香樟、枫香、海桐等),对疾病的控制有很强的现实意义。     

川中丘陵区柏木（Cupressus funebris）土壤浸提液对3种农作物种子萌发和幼苗生长的影响

某些植物和土壤浸提液的化感作用会影响种子萌发和幼苗生长.关于川中丘陵区的柏木(Cupressus funebris)土壤浸提液对作物种子萌发和幼苗生长是否有影响以及如何影响,推测柏木的土壤浸提液可能抑制作物的种子萌发和幼苗生长.采用室内种子萌发实验,研究土壤浸提液(0.00 g/mL、0.01 g/mL、0.02 g/mL、0.05 g/mL和0.10 g/mL)对番茄(Lycopersicon esculentum)、辣椒(Capsicum annuum)和茄子(Solanum melongena)种子萌发和幼苗生长的化感作用.结果表明,不同浓度土壤浸提液对作物种子萌发和幼苗生长有一定的影响,但与物种有关. 0.01 g/mL的土壤浸提液显著提高番茄的活力指数、降低根鲜重,0.05 g/mL的土壤浸提液显著增加番茄的根长,而0.10 g/mL的土壤浸提液显著降低番茄的发芽率. 4种浓度的土壤浸提液均显著降低辣椒的发芽势和苗鲜重,而辣椒的根长和根鲜重主要在0.01g/mL、0.02 g/mL和0.05 g/mL的土壤浸提液下降显著.对茄子而言,4种浓度的土壤浸提液均显著提高其活力指数、降低鲜根重.综合化感效应指数表明,柏木土壤浸提对番茄和茄子表现为抑制和促进作用,而辣椒仅表现为抑制作用,且3种作物的化感敏感性为辣椒茄子番茄.综上,柏木土壤浸提液能影响作物种子萌发和幼苗生长,且在川中丘陵区,辣椒可能不适合种植在以柏木为坡地防护林的坡耕地中.(图2表3参33)     

脆江蓠凝集素的部分性质及细胞凝集作用

脆江蓠藻体的磷酸盐缓冲液浸提液 ,经硫酸铵分级沉淀 ,再经DEAE -Sepharose和SephadexG 10 0层析纯化 ,得到脆江蓠凝集素 (GBL) .经测定该凝集素是一种分子量为 33.1× 10 3 ～ 33.8× 10 3 的糖蛋白 ,分子中含有 11%的中性糖 ,具有很高的耐热性 .GBL的凝血活性可被某些单糖或双糖及卵粘蛋白抑制 ,当浓度为 2 .34 μg/mL时引起兔红细胞凝集 ,浓度为 18.75 μg/mL时凝集绵羊红细胞 ,但对鸡、鸭、鸽子红细胞及人A、O、或B型血细胞没有凝集作用 .GBL能够凝集两种单细胞藻类 ,当浓度为 18.75 μg/mL时能凝集盐生杜氏藻 ,但藻细胞密度减少到 18.3× 10 4 /mL时不能发生凝集作用 ;GBL浓度为 9.37μg/mL时引起蛋白核小球藻凝集 ,但当藻细胞密度减少到 2 8.9× 10 4 /mL也不产生凝集反应 .GBL还能够凝集正常的枯草芽孢杆菌和经过热处理的酿酒酵母细胞 .图 3表 6参 14     

Role of bacteria in the epizootic ulcerative syndrome (EUS) of fishes

Bacteriological examination of certain water bodies and fishes carrying EUS was carried out. As a whole, 17 species of bacteria were isolated from the investigated water bodies and EUS affected fishes. The species of bacteria isolated from fishes are common to those isolated from water. Experimental infection trials conducted suggested that Aeromonas hydrophila in association with Pseudomonas fluorescens, may be playing the role of primary aetiological agent in producing EUS in fishes.     

海水网箱养殖大黄鱼细菌性病原鉴定与感染治疗研究

从患病大黄鱼的肌肉，肝、肾等病灶中分离出7种菌株，经人工感染试验证明，菌株970709A，970918a和970709B为致病菌，这三株菌株进行人工感染，死亡率均为100％，症状与自然发病相似，经菌体形态特征，生理生化特性反应测定，菌株970909A和970918a为鳗弧菌（Vibrio anguillarum)，菌株970909B为副溶血弧菌（V.parahaemolyticus)，药敏实验结果表明，氧氟沙星，环丙沙星，培氟沙星，罗红霉素，氯霉素和海鱼宁II等对致病菌有较强的抑制作用。临床治疗试验还表明，复方制剂海鱼宁II对大黄鱼弧菌病的治疗效果显著，有效率达66％－68％。     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Effect of neem extract against the bacteria isolated from marine fish

Marine ornamental fishes are exceedingly valuable due to their high demand in domestic and international markets. There is a growing global interest to rear the fishes in captivity. But problem due to bacteria and fungi are the major hitch in captive condition. Since, the use of antibiotics is banned, an attempt was made to ascertain in vitro assay of the neem leaves extract against the bacterial pathogens isolated from infected fishes. Bacterial strains isolated from infected regions of the clown fishes Amphiprion sebae and A. ocellaris were identified as Aeromonas hydrophila, Enterobacter sp., E. coli, Pseudomonas aeruginosa, Proteus sp., Streptococcus sp., Vibrio cholerae, V. alginolyticus, V. parahaemolyticus and Yersinia enterocolitica. Ethanol and methanol extracts were highly inhibitory to the bacterial isolates when compared to other solvents. Ethanol extracts exhibited low minimum inhibitory concentration (75-250 microg ml(-1)) as compared to other extracts. The present finding revealed that the neem leaf extract significantly reduces the bacterial pathogens and their infection in marine ornamental fishes.     

Innovative method of culturing bdelloid rotifers for the application of wastewater biological treatment

• An innovative method of culturing bdelloid rotifer fed on flour was proposed. • Rotifer fed on flour grew faster than that fed on bacteria or Chlorella vulgaris. • The optimum mass culture conditions for rotifer fed on flour were investigated. • The cultured rotifer could improve sludge settleability in the SBR. This study aims to establish a simple and efficient method for the mass culture of bdelloid rotifers, which is the basis for the application of bdelloid rotifers as biological manipulators to improve wastewater biological treatment performance. A common bdelloid rotifer, Habrotrocha sp., in a wastewater biological treatment system was selected as the culture target. Rotifers fed on flour could reproduce faster than those fed traditional food such as Chlorella vulgaris or mixed bacteria. As a rotifer food, flour has the advantages of simple preparation, effortless preservation, and low cost compared to live Chlorella vulgaris or mixed bacteria, so it is more suitable for the mass culture of rotifers. The optimal rotifer culture conditions using flour as food were also studied. According to the experimental results, the recommended rotifer culture conditions are a flour particle size of 1 μm, a flour concentration of 6 × 10⁶ cell/mL, a temperature of 28℃, a pH level of 6.5 and salinity of 100–500 mg/L. In addition, the sludge volume index in the sequencing batch reactor (SBR) with the addition of cultured rotifers was 59.9 mL/g at the end of operation and decreased by 18.2% compared to SBR without rotifer, which indicates that the cultured rotifers still retained the function of helping to improve sludge settling. This function may be related to the rotifer’s role in inhibiting bacteria from producing loosely bound extracellular polymeric substances in the SBR.     

铝对土壤微生物区系组成的影响

通过大豆盆栽试验,研究了金属铝对土壤微生物区系组成的影响。结果表明,当铝质量分数<1.1 g/kg时,土壤细菌、放线菌的数量随着铝质量分数的增加而增加;当铝质量分数>1.1 g/kg时,土壤细菌、放线菌的数量随着铝质量分数的增加而减少。铝对丝状真菌的生长具有明显的促进作用。在低质量分数下,铝对土壤氨化细菌、硝化细菌、反硝化细菌具有一定的刺激作用;当质量分数>1.1 g/kg时,则表现为抑制作用,最大抑制率分别达48%、96%和95%;土壤硝化细菌、反硝化细菌可以作为铝污染土壤生态环境的指示菌。自生固氮菌的数量随铝质量分数的升高而逐渐下降。铝对土壤微生物区系产生抑制作用的临界质量分数在1.3 g/kg左右。     

四种不同生态系统的土壤解磷细菌数量及种群分布

通过分析农田、林地、草地和菜地土壤有机磷细菌和无机磷细菌的数量及种群结构，发现有机磷细菌数量比无机磷细菌多；有机磷细菌主要是芽孢杆菌属，其次是假单胞菌属；而无机磷细菌主要是假单胞菌属。菜地土壤解磷细菌的数量和种类最多。不同的细菌解磷能力差异很大，其解磷能力与培养介质的ｐＨ值有一定的关系；许多菌株在进一步的纯化过程中失去了解磷能力。     

Effect of natural preservatives on the growth of histamine producing bacteria

Present study deals with the hampering of the growth of histamine producing bacteria (HPB), by using NaCl and spices which are easily available and cheaper cost wise. For this experiment, four strains of HPB viz. Vibrio parahaemolyticus, Bacillus cereus, Pseudomonas aeruginosa and Proteus mirabilis were tested against 1 to 10% concentrations of NaCl and 1 to 5% concentrations of natural preservatives (turmeric, ginger and garlic) in a basal medium. HPB showed different growth rates at different concentrations of NaCl and natural preservatives. V. parahaemolyticus, B. cereus and Ps. aeruginosa showed no growth at 10% concentration. When the HPB growth was tested with garlic, turmeric and ginger extracts, growth of all the bacteria was inhibited by garlic and turmeric extracts at 5% concentration. In ginger, V. parahaemolyticus, B. cereus and P. mirabilis were totally inhibited at 5% concentration. But Ps. aeruginosa showed very less growth at this concentration.     

越冬期淡水白鲳发病之病原细菌的研究

从越冬期淡水白鲳病鱼的肝脏内分离纯化到一细菌菌株ＣＢＺＨＧ４，用ｎ（ｃｅｌｌ＝１０＾５～１０＾６ｍＬ＾－１的菌液浓度在腹腔注射试验鱼后４８ｈ内全部死亡，证实为病原菌。ＣＢＺＨＧ４经ＡＰＩ ２０ＮＥ系统鉴定为嗜水性气单胞菌（Ａｅｒｏｍｏｎａｓ ｈｙｄｒｏｐｈｉｌａ），对卡那霉素多种抗生素和呋喃类药物敏感，对青霉素类、先锋霉素等抗生素、磺胺类药物和０／１２９不敏感。表２参２     

有机酸对珠江三角洲水稻土镉解吸行为的影响

用批平衡实验方法,研究了3种有机酸及其两两混合酸在一系列pH值梯度下(pH3~7)对珠江三角洲沉积型水稻土镉(Cd)解吸的影响.结果表明,只有柠檬酸及含柠檬酸的混合酸在较低酸度区(pH>5)可以显著促进Cd的解吸,这一效应与浓度成正相关,而苹果酸、草酸及二者的混合酸在所试验酸度范围均促进了Cd吸附,特别是草酸,促进作用随浓度的升高而增强.在较低pH区(pH<5),苹果酸最有利于Cd解吸,在高pH区(pH≥5),柠檬酸最有利于Cd解吸,草酸的Cd解吸能力最低.混合有机酸交互作用不显著,对Cd解吸的影响约等于各个单独酸的作用之和,但在一些酸度条件下,可存在颉颃作用.图3表2参22