微波诱变结合消泡剂定向选育VB12高产菌株的应用

运用微波诱变结合消泡剂定向选育VB12生产菌株——脱氮假单胞杆菌。结果表明：当微波功率800 W、脉冲频率2 450 MHz,分离平皿中消泡剂加量3‰时,诱变剂量6 s效果最明显。得到遗传稳定的突变菌株VB12Ⅰ09-1023#,通过摇瓶测得该突变株发酵单位高达268 mg/L,比出发菌株提高了28.8%。120 m3发酵罐试验,25批平均发酵单位高于对照7.5%。     

A preliminary study on the use of carbenicillin in snakes

The results of a 10 year survey on the in vitro antibiotic resistance patterns of Pseudomonas spp. isolated from clinically ill reptiles, showed a high degree of sensitivity to carbenicillin. On the basis of sensitivity testing, carbenicillin was used to treat nine snakes of four different species, with confirmed Pseudomonas infections. Plasma carbenicillin levels were assayed, by a microbiological agar diffusion technique, at intervals of time after a single intramuscular injection at a dose rate of 400 mg/kg. Peak plasma levels of 177 and 270 micrograms/ml were reached 1 h after the initial injection and therapeutic levels persisted for at least 12 h. This initial study indicated that a suggested dose regime in snakes, derived by extrapolation from mammalian dosages, of 100-125 mg/kg daily was insufficient to produce plasma levels of sufficient magnitude and duration to effectively treat Pseudomonas infections in snakes.     

优化VB12发酵工艺 提高经济效益

进行了甜菜碱不同加量对脱氮假单胞杆菌发酵产VB12影响的实验。结果表明,甜菜碱减量对VB12发酵单位不但无显著负影响,反而可以提高发酵单位。在此基础上考察了发酵培养基中甜菜碱不同加量对VB12发酵单位的影响,发酵培养基中甜菜碱的量由7．0g／L降至3．0g／L,对VB12发酵单位无显著影响;还考察了补料中甜菜碱减量对VB12发酵单位的影响,补料中甜菜碱量由30g/L降至15g/L,对VB12发酵单位无显著影响。     

水貂绿脓杆菌的分离与鉴定

2011~2013年间,从山东省、河北省和江苏省等地的多个貂场疑似水貂出血性肺炎病貂的肺脏、肝脏和脾脏中分离到425株疑似绿脓杆菌。用绿脓杆菌的OprF和PEA基因的PCR引物对分离菌株的目的基因扩增,经核苷酸测序确定其均为绿脓杆菌序列。通过日本生研株式会社血清学分型系统进行分型：G型376株,占88.5%;B型34株,占8%;C型12株,占2.8%;E型3株,占0.7%。取其中6株细菌（G血清型3株,其他血清型各1株）进行详细生化试验,结果显示该6株菌均符合绿脓杆菌的生化特性,并且从这6株中选4株细菌（每血清型各1株）腹腔接种小白鼠和气管内接种水貂,结果表明4株菌对其均有致病性。     

牛乳中嗜冷假单胞菌胞外蛋白酶的研究进展

假单胞菌通常被认为是危害原料乳及其制品的主要嗜冷菌,其分泌的胞外蛋白酶具有耐热性,甚至能在超高温灭菌处理后仍有活性残留,并减短超高温灭菌乳的货架期.本文对来自牛乳中的嗜冷假单胞菌蛋白酶的研究进展做了总结,重点假单胞菌蛋白酶的催化机制研究进展,相关基因包括蛋白酶基因apr X在假单胞菌鉴别中的应用研究进展.最后讨论了嗜冷假单胞菌蛋白酶需要深入研究的方向.     

抗鼻疽菌单克隆抗体的应用研究Ⅰ.免疫荧光间接法对鼻疽菌的检测

应用抗鼻疽菌单克隆抗体(McAb)作荧光抗体间接染色法代替习用的诊断血清检测鼻疽菌,证明McAb对鼻疽菌检测的敏感性高,特异性较强。     

Four-year survey of urinary tract infections in calves in Israel

The prevalence of urinary tract infections in calves aged seven days to three months in three dairy cattle herds ranged from 0.5 to 1.6 per cent, with an average of 1.1 per cent. The mortality rate reached 16.1 per cent. The morbidity rate of the female calves was 1.4 per cent and that of the male calves 0.8 per cent. The bacteria isolated from urine, and from vaginal and preputial swabs were Escherichia coli (35 per cent), Corynebacterium renale (14 per cent), plasma coagulase-negative Staphylococcus species (12 per cent), Pseudomonas aeruginosa (12 per cent), Proteus species (12 per cent) and Arcanobacterium pyogenes (5 per cent). The affected calves had a significantly lower serum concentration of inorganic phosphorus (P < 0.01).     

Research Progress on Biofilms in Pseudomonas aeruginosa

The opportunistic pathogen Pseudomonas aeruginosa is a gram-negative bacteria which can form biofilms.In this review,we summarized the biology mechanism of biofilms in Pseudomonas aeruginosa,including Pseudomonas aeruginosa adhesion,extracellular polysaccharide Psl and Pel,alginate participated in the biofilm maturation,and quorum sensing systems regulated the formation of biofilm in Pseudomonas aeruginosa,and the drug target on biofilm.The bacterial biofilms protected the bacteria from unsuitable environmental conditions.Through researching the structure and the pathogenesis of Pseudomonas aeruginosa,and its resistance molecular mechanisms,we could adjust or regulate the expression of biofilm-associated factors,optimize the anti-infection treatment in Pseudomonas aeruginosa.     

铜绿假单胞菌生物膜研究进展

条件致病菌铜绿假单胞菌（PA）是一种能形成生物膜的革兰氏阴性菌,作者综述了PA生物膜形成的生物学机制,包括菌体黏附、胞外多糖Psl和Pel、藻朊酸盐等参与细菌生物膜成熟的过程及群体感应系统调节相关因子表达,从而调控细菌形成生物膜应对不良环境。此外还概括了将生物膜作为靶点开发的药物等生物膜相关的研究进展。生物膜是菌体逃避有害刺激的护盾,研究其结构、形成及致病机理,了解PA产生耐药性的分子机制,对于通过调节生物膜形成或调控生物膜相关因子的表达进而优化PA的抗感染治疗有十分重要的意义。     

贵州修文猕猴桃溃疡病菌遗传多样性分析

采用16S rDNA序列分析及ERIC-PCR对从贵州修文贵长猕猴桃枝条、叶片、花蕾分离到的35株猕猴桃溃疡病菌进行遗传多样性分析。通过16S rDNA序列分析,鉴定所分离的病原菌为Pseudomonas syringae pv.actinidiae,并运用ERIC-PCR分析病菌群体的遗传多样性分析,相似系数为0.808时,35株菌株分为4个类群,85.7%的菌株属于第一个类群,且菌株无明显的采集地、采集部位的聚类;表明修文猕猴桃溃疡病菌具有丰富的遗传多样性;此研究为溃疡病的检测、防治提供科学依据。     

绿脓杆菌研究进展

绿脓杆菌存在于环境和正常畜禽体内,是一种条件致病菌。近年来,随着动物疾病研究的深入,有关绿脓杆菌感染的报道也越来越多。从病原学、流行病学、发病机理、临床学等方面对绿脓杆菌的研究进展进行了综述,以期为控制绿脓杆菌的感染奠定基础。     

恶臭假单胞杆菌转谷氨酰胺酶基因的克隆及原核表达载体的构建

通过PCR方法扩增恶臭假单胞杆菌(Pseudomonas putida)H76的转谷氨酰胺酶(transglutaminase,TGase)基因,然后将扩增的约800bp的基因片段克隆到pET-28a( )表达载体上,构建重组TGase的表达载体.酶切鉴定阳性的重组质粒命名为pET-TGase.核苷酸及推导的氨基酸序列分析表明,该基因与恶臭假单胞杆菌KT2440的相应基因有很高的同源性.该载体的构建为进一步研究转谷氨酰胺酶的生物学功能以及应用提供了基础.     

原料乳中耐冷假单胞菌的种类及其肽酶活力

在冷藏的原料乳中,假单胞菌经常出现并成为主导菌群,其分泌的耐热酶会使原料乳变质。对从原料乳中分离的102株假单胞菌经rpoB基因测序进行菌种分析,用偶氮酪蛋白测试所有假单胞菌在2种培养条件(30℃培养2d或6℃培养8d)下胞外肽酶的蛋白水解活性和耐热性。结果表明:荧光假单胞菌(Pseudomonas fluorescens)是假单胞菌中最主要的菌种,其次是莓实假单胞菌(Pseudomonas fragi)和假单胞菌sp1 (Pseudomonas sp1)。不同假单胞菌菌种之间的产肽酶能力存在明显差异。大多数荧光假单胞菌(23/41)在不同温度(30℃和6℃)条件下培养后能产肽酶,而大多数的假单胞菌sp1不产肽酶,所有分离的莓实假单胞菌(13/13)只在6℃条件下产肽酶,大多数的假单胞菌所产肽酶都有热稳定性。在6℃培养条件下,荧光假单胞菌菌株M02有最高肽酶活力(11.78ΔA/ (h·mL)),其胞外肽酶最佳催化pH值为6.5,最佳催化温度为40℃。当M02的接种浓度为103 CFU/mL时,6℃培养5d就能使脱脂乳溶液产生沉淀。对耐冷假单胞菌的种类和产肽酶能力进行分析和考察有助于提高原料乳或乳制品的质量和安全性。     

假单胞菌生物膜形成调控机制研究进展

假单胞菌是原料乳中常见的污染微生物,其会在乳品生产设备表面形成生物膜,导致设备腐蚀,且假单胞菌难以清除。同时假单胞菌形成的生物膜能促进假单胞菌耐热酶的分泌,还可以作为致病菌的生长基质,从而给乳品安全带来风险。本文总结了假单胞菌生物膜调控机制的研究进展和影响生物膜形成的环境因素,为假单胞菌生物膜的防控提供依据。     

Evaluation of activity of selected ophthalmic antimicrobial agents in combination against common ocular microorganisms

OBJECTIVE: To determine in vitro efficacy of gentamicin, tobramycin, and miconazole when used in combination, with or without atropine, against Pseudomonas or Aspergillus sp. PROCEDURE: Selected ophthalmic agents were combined for predetermined times. Sterile disks impregnated with the combined solutions were prepared and placed on Mueller-Hinton plates that were seeded with Pseudomonas or Aspergillus sp. Zones of growth inhibition were measured at postincubation hours 24 and 48. RESULTS: Tobramycin alone inhibited growth of Pseudomonas sp, whereas miconazole inhibited growth of Aspergillus sp. Significant differences in zones of growth inhibition when atropine was combined with tobramycin, when gentamicin was combined with miconazole, or when atropine was combined with miconazole and gentamicin, were not detected. CLINICAL RELEVANCE: Combining selected ophthalmic therapeutic agents for as long as 6 hours does not appear to alter the in vitro efficacy of the agents against microorganisms used in this study.     

Long-term cryopreservation of microbial strains relevant to food hygiene

Experiments on long-time cryopreservation at -20 degrees C were conducted, using 42 microbial strains of relevance to food hygiene. Blood slant agar was coated with 10% glycerol solution, so that all strains were in re-culturing conditions, following 12 months of storage. Re-culturing of 18 in 20 of the microbial genera used in the experiment was possible after 24 months of storage, if strains had been kept only in 10% glycerol solution. The method proved suitable particularly for strain keeping of highly sensitive genera (Lactobacillus, Pseudomonas, Aeromonas, Shigella).     

Variability of laboratory identification and antibiotic susceptibility reporting of Pseudomonas spp. isolates from dogs with chronic otitis externa

The purpose of this study was to evaluate interlaboratory variation in isolation and antibiotic susceptibility pattern of Pseudomonas spp. as reported to veterinarians for cases of canine chronic bacterial otitis externa. Twenty-six dogs with unilateral or bilateral bacterial otitis externa from multiple referral practices were included in this prospective study. Triplicate samples collected simultaneously from the same location in the external ear canal were randomly submitted to three laboratories for culture and susceptibility testing. Pseudomonas spp. were isolated from 18 of 34 (53%) ears. All three laboratories agreed on the presence of Pseudomonas spp. in 15 (83.3%) ears sampled. However, two laboratories agreed on two (11.1%) occasions, and on one occasion (5.5%) Pseudomonas spp. were identified in only one laboratory. Minimum inhibitory concentration (MIC) susceptibilities to 11 antibiotics were compared between laboratories B and C. Using laboratory-defined susceptibility of sensitive (S), intermediate (I) and resistant (R), none of the 16 Pseudomonas spp. with MIC data reported had identical patterns of antibiotic susceptibility. Agreement in susceptibility to individual antibiotics was observed in 13 of 16 (81%) occasions for amikacin and gentamicin, 10 of 16 (63%) occasions for ticarcillin, and nine of 16 (56%) for enrofloxacin. These results indicate that Pseudomonas spp. were identified by all three laboratories chosen for this study in 83% of the time. Moreover, antibiotic susceptibility patterns and MIC values reported to veterinarians may not agree between laboratories. Veterinarians should interpret bacterial culture and susceptibility results with multiple caveats including variability between laboratories.     

绿脓杆菌焦磷酸测序检测方法的建立

本试验旨在利用绿脓杆菌的序列信息分析和焦磷酸测序技术,建立一种快速、简单检测绿脓杆菌的方法。从培养的绿脓杆菌中提取DNA,PCR扩增目的基因片段,采用焦磷酸测序技术(pyrosequencing technology,PSQ)针对保守核苷酸区段的测序分析。通过焦磷酸测序后获得的序列信息与已知的目的基因的序列比对,能进一步确证毒株的序列信息为绿脓杆菌。利用焦磷酸测序能快速获取核酸的序列信息,可为毒株及早确证奠定基础。     

不同桑品种对桑青枯病抗性鉴定

通过疫区栽植、水培投菌、土壤浇菌等接种法,对50多个桑品种进行桑青枯病抗病性鉴定试验,调查结果表明了不同桑品种间对桑青枯病的抗性有明显差异。结果,白皮大种、真杜子桑、育2号、湖桑39等4个具有较高的抗病能力的桑品种可作为育种材料,并可在生产上利用。     

一例雏鸡铜绿假单胞菌的分离鉴定及其16SrRNA基因序列分析

为对死亡雏鸡进行病因诊断,通过大体剖检、细菌分离、生化鉴定,证实为铜绿假单胞菌感染。测定了该分离株的16SrRNA基因序列,并与GenBank中收录的序列比较,结果发现所分离的铜绿假单胞茵及参考株的16SrDNA基因序列极其保守,相似性达99％～100％;与大肠杆菌、沙门氏菌、巴氏杆菌的相似性差异约为9％;与鸭疫里默氏...