金樱子茎提取物体外抑菌活性研究

采用纸片扩散法和试管稀释法,研究金樱子茎不同溶剂粗提取物对临床常见病原细菌的抑菌活性。结果表明,金樱子茎水提取物对绿脓杆菌、痢疾杆菌、大肠杆菌、变形杆菌无抑菌活性;对伤寒杆菌的最低抑菌浓度（MIC）为20 mg（生药）/mL;对金黄色葡萄球菌的MIC为10 mg（生药）/mL。金樱子茎75%乙醇提取物对绿脓杆菌、伤寒杆菌、痢疾杆菌、大肠杆菌、变形杆菌无抑菌活性;对金黄色葡萄球菌MIC为10 mg（生药）/mL。金樱子茎乙酸乙酯、氯仿、石油醚提取物对绿脓杆菌、伤寒杆菌、痢疾杆菌、大肠杆菌、金黄色葡萄球菌、变形杆菌无抑菌活性。     

20种中药提取物对牙龈卟啉单胞菌抑制作用的体外研究

目的筛选评价20种中药提取物对牙龈卟啉单胞菌抑制作用。方法选择20种有较强抑菌作用及清热解毒作用的中药提取物,采用杯碟法进行体外抑菌试验,比较各中药对牙龈卟啉单胞菌的抑菌环直径和最低抑菌浓度(MIC)。结果水煎剂中黄连、乌梅、五味子、五倍子和黄芩的MIC为0.0488~1.5625mg/mL。油剂中丁香油、香薷油的MIC为0.3906~1.5625mg/mL。结论本实验选择的20种中药提取物中香薷油、黄连、乌梅、五味子、五倍子和黄芩的抑菌敏感度最高。     

茶叶水提物对猪源耐药大肠杆菌的抑制作用

本研究旨在考察茶叶水提物对耐药大肠杆菌的抑制作用,为大肠杆菌病的防治提供依据。采用体外抑菌试验测定红茶水提物和绿茶水提物对猪源耐药大肠杆菌分离株的抑制活性,然后观察具有较强抑菌活性的绿茶水提物对大肠杆菌攻毒小鼠的保护作用。结果表明,随着浓度的增加,两种茶叶水提物的抑菌活性增强;红茶水提物和绿茶水提物对13株大肠杆菌分离株的MIC值分别为62.5~250.0 mg/m L、62.5~125.0 mg/m L,提示绿茶水提物的抑菌活性较强;两种茶叶水提物对13个大肠杆菌分离株的抑制活性强弱表现出一定的差异。攻毒后48 h,攻毒组小鼠全部死亡,绿茶水提物组小鼠的存活率为20%;绿茶水提物组小鼠的十二指肠和空肠肠绒毛高度与对照组比较差异不显著,而显著大于攻毒组(P0.05),提示绿茶水提物对大肠杆菌攻毒小鼠的肠道损伤具有一定缓解作用。上述结果提示,绿茶水提物对大肠杆菌具有很好的抑制作用,可作为防治大肠杆菌病的备选生物活性物质之一。     

链霉菌H03发酵液提取物的抗菌活性研究

以常见的病原细菌和植物病原真菌为指示菌研究了链霉菌发酵液提取物的抗菌活性,测定了发酵液提取物的抗菌谱和最小抑菌浓度(MIC)。结果表明:与青霉素、链霉素相比,该菌发酵液提取物的抗菌范围更广,对植物病原真菌棉花黄萎病菌、苹果轮纹病菌、小麦赤霉病菌、油菜菌核病菌、黄瓜炭疽病菌、甜菜褐斑病菌、稻瘟病菌,以及常见病原细菌大肠杆菌、枯草杆菌、绿脓杆菌和金黄色葡萄球菌都具有明显的抑制作用。利用试管二倍稀释法测定发酵液提取物对上述诸菌的最小抑菌浓度(MIC),结果分别为0.125、0.062 5、0.125、0.25、0.015、0.03、0.015、0.015、0.03、0.250、.015 mg/mL。其中对植物病原真菌中的黄瓜炭疽病菌、甜菜褐斑病菌、稻瘟病菌,以及病原细菌中的大肠杆菌、枯草杆菌、金黄色葡萄球菌抑制效果最佳。将发酵液提取物置于100℃下加热处理10 min后,其对金黄色葡萄球菌的抑菌活性不变,说明该发酵液提取物具有较好的热稳定性。     

对中药提取物体外抗菌活性的研究

研究中药提取物对体外抗菌活性的抑制作用可以采用的方法主要有琼脂平板培养计数法和肉汤二倍稀释法。在本实验中采用的菌类有沙门氏菌、巴氏杆菌和大肠杆菌。实验数据显示,在七种中药材中,抗菌活性各不相同,最强的是黄柏提取物,其最小杀菌浓度(MBC)在31.25~250 mg.m L-1范围内,最小抑菌浓度(MIC)在15.6~125mg.m L-1范围内;抗菌活性强度排在第二位的是栀子提取物,MBC为125~250 mg.m L-1,MIC为62.5~125 mg.m L-1;抗菌活性相对更稳定的是佛手提取物,MBC为125 mg.m L-1和250 mg.m L-1,MIC均为125 mg.m L-1;抗菌活性相对较差的是何首乌、姜黄、细毡毛忍冬和杜仲的提取物,大部分MIC大于等于250 mg.m L-1。研究结论为,四川地区的的七种中药提取物中,有进一步研究价值的是黄柏、栀子和佛手。     

山豆根甲醇提取物对三七重要病原真菌的抑菌研究

利用超声波辅助甲醇提取山豆根粉末得到甲醇提取物;以3种三七病原真菌为靶标菌,采用菌丝生长法和肉汤稀释法,测定其抑菌活性。结果表明,该提取物得率为10.34%;其对3种病原真菌的EC_(50)、EC_(90)、EC_(95)分别为0.793~1.214mg/mL、6.046~6.432mg/mL、9.61~11.643mg/mL,约为阳性对照的2至8倍;其对3种病原真菌生长的最小抑菌浓度均为2.5mg/mL,为阳性对照的5至10倍。山豆根甲醇提取物对3种三七病原真菌均有强的抑菌活性,具有较大的开发应用潜力。     

患病大鲵中弗氏柠檬酸杆菌的分离与鉴定

【目的】确定导致大鲵(Andrias davidianus)细菌性感染死亡的病原。【方法】从大鲵肝脏中分离细菌,通过Biolog微生物自动鉴定系统及分子生物学方法对纯培养的细菌进行鉴定,再用大鲵和鲫鱼分别进行人工感染试验,以确定分离菌的致病性,同时对分离到的病原菌进行药物敏感试验。【结果】从患病大鲵肝脏中分离到一株致病菌JZ01,经人工感染健康大鲵,可复制与自然发病相同的症状,且从人工感染病鲵体内再次分离到相同的病原菌。该致病菌对健康鲫鱼也有致病性。经Biolog微生物自动鉴定系统的鉴定,以及进一步的16S rDNA基因序列和系统发育分析都表明,此致病菌为弗氏柠檬酸杆菌。药物敏感性试验表明,该菌株对氨曲南、头孢三嗪、先锋噻肟等9种药物高度敏感。【结论】弗氏柠檬酸杆菌是大鲵的一种致病菌。本文在国内外首次报道了该菌对大鲵具有致病性。     

苦参、黄芩、乌梅对金黄色葡萄球菌、大肠埃希菌和白假丝酵母菌抗菌活性的研究

目的观察苦参、黄芩和乌梅的体外抗菌活性。方法采用M-H琼脂连续稀释法做抗菌作用测定,测出各中药的最小抑菌浓度。结果苦参对金黄色葡萄球菌的MIC值最低（2.5mg/ml）,乌梅对大肠埃希菌的MIC值最低（1.25mg/ml）,乌梅对白假丝酵母菌的MIC值最低（2.5mg/ml）。结论苦参、黄芩和乌梅对革兰阳性球菌、革兰阴性杆菌和真菌均有较好的抑菌活性。     

湘西虎杖抑菌活性成分提取研究

对湘西虎杖抑菌活性物提取条件进行研究,通过抑菌实验得到最佳提取条件为90%乙醇、60℃、3 h,该条件下的提取物对细菌生长的抑制活性最强,但对酵母菌无明显的效果,对金黄色葡萄球菌、大肠杆菌、痢疾杆菌的最小抑菌浓度（MIC）为0.25 g/mL～0.5 g/mL.提取物经85℃、100℃,15 min或pH 2～8处理后仍具有较强的抑菌活性.     

墨旱莲提取物抑菌活性初步研究

对药用植物墨旱莲Eclipta prostrata提取物的抑菌活性物质进行研究。结果表明：（1）通过不同部位提取物的抑菌试验,发现只有叶和根茎含有抑菌活性物质;（2）墨旱莲提取物对金黄色葡萄球菌、大肠杆菌、枯草杆菌、藤黄球菌的最低抑菌浓度分别为0.6、2.0、0.8、1.0 g/mL,最小杀菌浓度均为1.0 g/mL,对白色念珠菌有很强的抑菌作用,但对黑曲霉无效;（3）通过不同极性溶剂的抑菌试验表明,抑细菌活性物质易溶于石油醚、乙醚等非极性溶剂,具有最佳抑细菌效果的提取溶剂为偏碱性的65%乙醇溶液;（4）墨旱莲的抑菌活性物质可能为黄酮类物质。     

Pathogenicity of pap-negative avian Escherichia coli isolated from septicaemic lesions

Recent studies by DNA-DNA hybridisation assays conducted on a large collection of Escherichia coli strains isolated from chickens, ducks and turkeys suffering from colibacillosis, showed that 76% of the strains were negative for the presence of the pap gene cluster. The objective of this paper was to study the virulence associated with the avian E. coli strains negative for the P fimbriae, but carrying the f17 or the afa-8 gene cluster coding for adhesins associated with strains pathogenic for mammals. Three strains carrying the f17 fimbriae and three carrying the afa-8 adhesin-encoding gene cluster were studied in three in vivo experimental models of avian colibacillosis: subcutaneous inoculation of 1-day-old chicks, inoculation of specific-pathogen-free (SPF) chickens via the intra-thoracic air sac, and intra-tracheal inoculation of axenic chickens. The results showed that the six P-negative E. coli isolates carrying the f17 or the afa-8 gene cluster were lethal for 1-day-old chicks. They were also able to reproduce clinical signs and lesions of colibacillosis (aerosacculitis, pericarditis, perihepathitis), with bacteraemia and septicaemia, in SPF chickens inoculated via the thoracic air sacs as well as in axenic chickens inoculated by the intra-tracheal route. Further studies with f17 and afa-8 allelic mutants constructed by disruption must be performed to confirm a role of F17 fimbrial and Afa-VIII afimbrial adhesins in the pathogenesis of avian colibacillosis.     

中药水提物对白念珠菌生物膜抑制作用的研究

目的研究中药水提物对体外白念珠菌生物膜的影响。方法体外构建白念珠菌生物膜,微量稀释法测定中药提取物对白念珠菌浮游菌最低抑菌浓度（M IC）,XTT减低法评价中药提取物对白念珠菌生物膜SM IC（SM IC80,SM IC50）及对白念珠菌黏附能力的影响。结果野菊花、土槿皮和决明子SM IC80为125 mg/m l,其余中药SM IC80为250或500 mg/m l;地肤子SM IC50为7.81 mg/m l,川芎、苏梗、藿香和决明子SM IC50为15.63 mg/m l,其他中药SM IC50在62.5~250 mg/m l。10味中药在100 mg/m l时均能显著抑制白念珠菌细胞的黏附,10 mg/m l时地肤子、苏梗和苦参对黏附仍具抑制作用。结论该10味中药水提物对体外白念珠菌生物膜有较强抑制效应。     

Comparison of extraintestinal pathogenic Escherichia coli strains from human and avian sources reveals a mixed subset representing potential zoonotic pathogens

Since extraintestinal pathogenic Escherichia coli (ExPEC) strains from human and avian hosts encounter similar challenges in establishing infection in extraintestinal locations, they may share similar contents of virulence genes and capacities to cause disease. In the present study, 1,074 ExPEC isolates were classified by phylogenetic group and possession of 67 other traits, including virulence-associated genes and plasmid replicon types. These ExPEC isolates included 452 avian pathogenic E. coli strains from avian colibacillosis, 91 neonatal meningitis E. coli (NMEC) strains causing human neonatal meningitis, and 531 uropathogenic E. coli strains from human urinary tract infections. Cluster analysis of the data revealed that most members of each subpathotype represent a genetically distinct group and have distinguishing characteristics. However, a genotyping cluster containing 108 ExPEC isolates was identified, heavily mixed with regard to subpathotype, in which there was substantial trait overlap. Many of the isolates within this cluster belonged to the O1, O2, or O18 serogroup. Also, 58% belonged to the ST95 multilocus sequence typing group, and over 90% of them were assigned to the B2 phylogenetic group typical of human ExPEC strains. This cluster contained strains with a high number of both chromosome- and plasmid-associated ExPEC genes. Further characterization of this ExPEC subset with zoonotic potential urges future studies exploring the potential for the transmission of certain ExPEC strains between humans and animals. Also, the widespread occurrence of plasmids among NMEC strains and members of the mixed cluster suggests that plasmid-mediated virulence in these pathotypes warrants further attention.     

诃子水提物对泛耐药鲍曼不动杆菌的抑菌作用

摘要：目的 研究诃子水提物对泛耐药鲍曼不动杆菌（PDR-Ab）的抑菌作用。方法 采用二倍稀释法测定诃子水提物对临床分离的泛耐药鲍曼不动杆菌的最低抑菌浓度（MIC）和最低杀菌浓度（MBC）。小鼠腹腔注射上述PDR-Ab,建立急性肺炎模型。于造模前4天至造模后2天对小鼠连续灌胃给药,考察小鼠的死亡率、血液白细胞计数及肺组织病理变化。结果 体外实验表明诃子水提物能抑制PDR-Ab在液体培养基中的生长,MIC和MBC分别为0.75 mg/mL和12 mg/mL。体内实验表明,剂量为150 mg/kg诃子水提物灌胃给药能明显降低PDR-Ab感染性急性肺炎小鼠的死亡率,使血液白细胞计数较快恢复,肺组织病理改善明显。结论 诃子水提物对PDR-Ab在体内外均具有明显的抑菌作用,有望用于PDR-Ab感染性疾病的治疗。     

Genotypic analyses of Escherichia coli isolated from chickens with colibacillosis and apparently healthy chickens in Japan

A genotypic comparison using pulsed-field gel electrophoresis (PFGE), amplified ribosomal restriction analysis (ARDRA) as well as PCRs targeting virulence associated genes reported elsewhere in avian pathogenic Escherichia coli(APEC) was made between E. coli strains isolated from chickens with colibacillosis and those from the feces of apparently healthy chickens in Japan. The majority (67%) of clinical isolates belonged to a certain phylogenetic ARDRA but not PFGE cluster, with virulence-related genes carried by ColV plasmid being markedly prevalent. The result suggests that APEC strains originated from the same "ancestor" in the course of E. coli evolution.     

Antimicrobial activity of organic extracts isolated from Aplysina fistularis (Demospongiae: Aplysinidae)

Organic extracts of the sponge Aplysina fistularis (Pallas 1766) were tested for antimicrobial activity against Gram positive bacteria (Staphylococcus aureus) and Gram negative bacteria (Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa). The minimal inhibitory concentration (MIC) and toxic activity of extract were determined. Susceptibility trials of organic fractions obtained by VLC: Hexane, EtOAc and CHCl3 showed that EtOAc fraction has antibacterial activity against E. coli, while CHCl3 fraction inhibited E. coli and S. aureus growth. The later refractioning of EtOAc fraction and the biodirected assays showed that fractions F12 and F13 of EtOAc/Hex and EtOAc F14 were bioactive against Gram positive and Gram negative bacteria. Only EtOAc/MeOH Sf2 from subfractionig of EtOAc F14 produced inhibition for E. coli and S. aureus. In Sf2 EtOAc/MeOH, MIC was moderate for S. aureus (MIC > 256 g/ml). F4 CHCl3/MeOH produced a high inhibition in S. aureus (MIC = 0.125 g/ml) and for E. coli (MIC > 16 g/ml). F10 CHCl3/MeOH showed a moderate activity against S. aureus (MIC > 128 g/ml) and low activity against E. coli (MIC = 512 g/ml). F10 CHCL3/MeOH did no present toxic activity against Artemia salina. The fractiorts F4 CHCL3/MeOH and Sf2 EtOAc/MeOH were toxic for this organism when the concentration was higher than 100 microg/ml. LC50 in both cases was 548.4 and 243.4 microg/ml respectively. Secondary metabolites of medium polarity obtained from A. fistularis have a wide spectrum of anti bacterial activity. Toxicity analysis suggests that only F10 CHCL3/MeOH has potential as an antimicrobial agent for clinical use.     

Detection of Iss and Bor on the surface of Escherichia coli

AIMS: To confirm the presence of Iss and Bor on the outer membrane of Escherichia coli using Western blots of outer membrane protein (OMP) preparations and fluorescence microscopy, and explore the use of fluorescence microscopy for the detection of avian pathogenic E. coli (APEC) and diagnosis of avian colibacillosis. METHODS AND RESULTS: Knockout mutants of iss and bor were created using a one-step recombination of target genes with PCR-generated antibiotic resistance cassettes. Anti-Iss monoclonal antibodies (Mabs) that cross-react with Bor protein were used to study the mutants relative to the wild-type organism. These Mabs were used as reagents to study OMP preparations of the mutants with Western blotting and intact E. coli cells with fluorescence microscopy. Iss and Bor were detected in Western blots of OMP preparations of the wild type. Also, Iss was detected on Deltabor mutants, and Bor was detected on Deltaiss mutants. Iss and Bor were also detected on the surface of the intact, wild-type cells and mutants using fluorescence microscopy. CONCLUSIONS: These results demonstrate that Bor and Iss are exposed on E. coli's outer membrane where they may be recognized by the host's immune system. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report confirming Iss' location in the outer membrane of an E. coli isolate. Such surface exposure has implications for the use of these Mabs for APEC detection and colibacillosis control.     

Evaluation of phytochemical and antimicrobial properties of leaf extract of Tapinanthus sessilifolius (P. Beauv) van Tiegh

Leaf extracts of T. sessilifolius growing on five different host plants (Psidium guajava, Citrus lemon, Vernonia amygdalina, Persea americana and Jatropa curcas) were evaluated for antimicrobial activity of the plant. Powdered leaves of T. sessilifolius collected from each host plant was divided into two portions. One portion was used for aqueous infusion and the other portion was successively extracted with hexane, ethylacetate and methanol. Infusion of aqueous extract of powdered leaves did not show antimicrobial effect even at the concentration of 1000 and 2000 microg/ml on test microorganisms (Staph. aureus, E. coli, Bacillus subtilis, Pseudomonas aeruginosa and Candida albicans). However in broth culture, methanolic and hexane extract had MIC range of 62.5-500 microg/ml and ethylacetate extract had 250-500 microg/ml. Phytochemical screening of leaf samples of T. sessilifolius collected from different host plants showed positive test for hydrolysable tannins, saponins, flavonoids, terpenes, cardiac glycoside, reducing sugars and proteins. LD50 concentration was found to be > 1.500 mg/kg for samples from P. guajava; 489.89 mg/kg for J. curcas and C. lemon; and 692 mg/kg for V. amydalina in mice.     

Characterization of the coliform and enteric bacilli in the environment of calves with colibacillosis

In the first part of the present study the coliform and enteric bacilli in the environment of calves with colibacillosis were examined. The occurrence, number, and pathogenic properties of Escherichia coli in barnyard soils were obtained from six cattle ranches. The O and K serogroups of E. coli isolates obtained from the feces of calves with colibacillosis born at these cattle ranches were determined, and their serotypes were compared with the E. coli O and K serotypes found in soils. The results showed a reservoir of potentially pathogenic E. coli in barnyard soils contaminated with bovine feces. For the second part of this study, 6 healthy calves and 51 calves with colibacillosis were studied. The numbers of total aerobic heterotrophic bacteria, total streptococci, fecal streptococci, total coliforms, and fecal coliforms in the feces of calves were determined. In addition, coliform and enteric bacilli from the feces of both healthy and diseased calves were identified, and their indole, methyl red, Voges-Proskauer, citrate (IMViC) types were described. In parallel, the IMViC types of coliform and enteric bacilli isolated from barnyard soils previously contaminated with bovine feces were compared with those isolated from uncontaminated soils. All fecal specimens were also examined for the presence of rotavirus. No significant effect on the numbers of the bacterial types was found. The results suggest that the predominant IMViC types found in the feces of calves with colibacillosis originate from the soil. From this study it is apparent that the occurrence, number, and survival of E. coli in barnyard soils is related to ranch husbandry and sanitary practices.     

Characterization of the coliform and enteric bacilli in the environment of calves with colibacillosis.

In the first part of the present study the coliform and enteric bacilli in the environment of calves with colibacillosis were examined. The occurrence, number, and pathogenic properties of Escherichia coli in barnyard soils were obtained from six cattle ranches. The O and K serogroups of E. coli isolates obtained from the feces of calves with colibacillosis born at these cattle ranches were determined, and their serotypes were compared with the E. coli O and K serotypes found in soils. The results showed a reservoir of potentially pathogenic E. coli in barnyard soils contaminated with bovine feces. For the second part of this study, 6 healthy calves and 51 calves with colibacillosis were studied. The numbers of total aerobic heterotrophic bacteria, total streptococci, fecal streptococci, total coliforms, and fecal coliforms in the feces of calves were determined. In addition, coliform and enteric bacilli from the feces of both healthy and diseased calves were identified, and their indole, methyl red, Voges-Proskauer, citrate (IMViC) types were described. In parallel, the IMViC types of coliform and enteric bacilli isolated from barnyard soils previously contaminated with bovine feces were compared with those isolated from uncontaminated soils. All fecal specimens were also examined for the presence of rotavirus. No significant effect on the numbers of the bacterial types was found. The results suggest that the predominant IMViC types found in the feces of calves with colibacillosis originate from the soil. From this study it is apparent that the occurrence, number, and survival of E. coli in barnyard soils is related to ranch husbandry and sanitary practices.