病原体致病性和毒力的现代概念

微生物致病性和毒力的传统概念不能反映病原体与宿主相互作用的复杂关系。本文介绍一种损伤－应答分类法,将致病性的现代概念。     

病原体毒力的进化

病原体毒力的进化是进化生物学和传染病预防控制中的重要课题。最具有进化优势的病原体应该是能够最大限度传播自身,对宿主造成的危害越小,越有利于病原体的长期传播。但根据“交易模型”,病原体在具有传播能力的同时,不可避免地造成宿主适应性的降低。由这一模型预测的病原体毒力进化,会在传播率最大和对宿主影响最小之间取得平衡,以便病原体的基本繁殖率最大化。本文介绍了近年来对这一模型的研究结果、其可能的使用意义和存在的争议,以及进一步完善模型的必要性．     

疫苗类型与病原体毒力演变的关系

Gandon通过建立病原体系力演变数学模型,研究有缺陷疫苗对病原体毒力演变的影响。发现4种类型疫苗中抗生长疫苗,抗毒疫苗使用后会导致原体演变为高毒力株,抗感染疫苗使用后可使原体演变为低毒力株,抗传播疫苗对病原体毒力影响较小,这一发现有助于疫苗的研究和应用。     

女性生殖道感染常见病原体及药敏研究进展

目的 生殖道感染在妇科疾病中发病率较高,可造成不孕、异位妊娠、胎膜早破、早产等不良后果。引起女性生殖道感染的病原体种类较多,且由于临床不合理用药现象日趋严重,各种病原体对抗菌药物的耐药性逐年上升。本文对引起女性生殖道感染的常见病原体种类、病原体对女性生殖健康的影响及药敏试验研究进展进行了综述。了解各病原体特点,开展各种病原体对药物敏感性试验,可减少女性生殖道感染的复发率,提高治愈率,为改善女性生活质量与促进生殖健康提供帮助。     

嗜肺军团菌毒力基因、致病性及实验诊断研究进展

嗜肺军团菌是引起军团菌肺炎的重要病原体,关于军团菌感染的致病机制目前尚不十分清楚。近来研究发现,军团菌的致病性与其毒力因子和铁代谢等相关,本文对嗜肺军团菌的毒力因子及其作用。致病机制和实验室诊断方法研究进展作一综述。     

莱姆病的实验诊断

本文简要地说明一些可供莱姆病实验诊断的方法及其应用价值,包括培养分离和组织中的直接检测、免疫学方法和分子生物学方法,并对各种方法的应用价值进行了评估。     

复合性感染的复数菌药敏实验分析

从检材中培养出细菌是确立感 (传 )染性疾病诊断和治疗的基本依据 ,单一需氧菌的培养方法易遗漏厌氧菌和缺壁细菌 ,不能概括致病原因菌的全貌 [1 ] ,使多种类细菌的混合性感染不能准确诊断和治疗。我室于 1999年接待了用常规方法仅培养部分需氧菌或未培养出细菌的检材 3例 (血 1例、腹水 1例、尿 1例 ) ,同时进行需氧菌、厌氧菌及细菌 L型培养 ,检测结果报告如下 :1　材料与方法1.1　标本来源　标本来自本市医院患儿 3例 ,拟诊为“败血征”血 1例 ,诊断为“脾亢”腹水 1例 ,诊断为“膀胱炎”尿 1例。1.2　培养基1.2 .1　需氧菌 (简称 X)培…     

细鳞鲑细菌性败血症病原菌的分离鉴定及药敏实验

从21尾患病细鳞鲑Brachymystax lenok体内分离到1株致病菌,经分离培养、生化鉴定、16S r DNA序列分析及人工感染实验,确定该病原菌为嗜水气单胞菌Aeromonas hydrophila。采用26种抗菌药物进行药敏分析,结果发现,分离菌株对氨苄西林、头孢呋辛钠、卡那霉素等10种抗生素敏感;对羧苄西林、头孢噻肟、环丙沙星、新霉素等9种抗生素中度敏感;对大观霉素、诺氟沙星、四环素等7种药物耐受,为细鳞鲑细菌性疾病的防控提供了重要依据。     

烟曲霉抗原及实验诊断的研究进展

烟曲霉是多种医源性因素导致的机会性,严重深部真菌感染的主要病原菌,烟曲霉病的早期诊断及防治是亟待解决的问题,目前,研究烟曲霉特异性抗原是发展早期诊断曲霉病方法的基础。     

Effect of oral immunization with Aeromonas hydrophila ghosts on protection against experimental fish infection

Aims:  To investigate whether oral immunization with Aeromonas hydrophila ghosts (AHG) vaccine can elicit mucosal and systemic immune responses of Carp ( Carassius auratus gibelio ) compared to conventional formalin-killed bacteria (FKC).
Methods and Results:  Fish were fed diets coated with AHG, FKC or phosphate buffered saline (PBS) alone, after immunization, more antigen-specific antibody was significantly detected in serum and intestinal mucus in AHG group than FKC group and PBS group. In addition, after challenged with the parent strain J-1, the survival of bacterial ghost-vaccinated fish was higher than PBS group and FKC group, the relative per cent survival (RPS) being 76·8%, 58·9%, respectively.
Conclusions:  Oral immunization with A. hydrophila ghosts can elicit systemic and mucosal adaptive immune responses and has higher potential to induce protective adaptive immunity than normal vaccine.
Significance and Impact of the Study:  Oral immunization with bacterial ghosts is a promising new solution with potential application to prevent diseases in fish.     

Properties of membranous phospholipase C from WRK1 cell: sensitivity to guanylnucleotides and bacterial toxins

As previously described, WRK₁ plasma membrane possesses a vasopressin-sensitive phospholipase C [G. Guillon et al., FEBS Lett. 196, 155–159]. In the present study, we examined the sensitivity of this enzyme to guanylnucleotides. GTPγS induced a time- and dose-dependent stimulation of Ins(1,4,5)P₃ and Ins(1,4)P₂ accumulation. No accumulation of InsP₁, Ins(1,3,4)P₃ or Ins(1,3,4,5)P₄ occured under similar conditions. Gpp(NH)p produced the same effect but was less potent. GTP and a nonhydrolyzable analogue of ATP, App(NH)p, were without effect. Calcium also stimulated the phospholipase C activity in a time- and dose-dependent manner. In the absence of calcium, the activity of GTPγS was considerably reduced. Physiological calcium concentrations (between 10⁻⁸ and 10⁻⁷M), allowed maximal GTPγS stimulation of phospholipase C activity. In this system, the presence of vasopressin alone did not generate inositol phosphate accumulation. However, this hormone: (i) reduced the lag-time observed during GTPγS stimulation, (ii) increased the sensitivity of phospholipase C to GTPγS, and (iii) did not modify the stimulation of phospholipase C induced by maximal doses of GTPγS. Unlike sodium fluoride, GTPγS elicited an irreversible activation of phospholipace C. Calcium, GTPγS and sodium fluoride stimulated the phospholipase C activity via mechanisms sharing a common step, since their maximal effects were not additive. Cholera toxin treatment, known to produce complete ADP-ribosylation of ‘s’ subunits, partially reduced the basal and the maximal GTPγS-mediated stimulation of phospholipase C activity as well as that caused by vasopressin. This inhibition was not mimicked by treatment with either forskolin or pertussi toxin.     

Isolation and characterization of bacteriophage PM3 from Aeromonas hydrophila the bacterial receptor for which is the monopolar flagellum

PM3 is an Aeromonas-specific bacteriophage which was isolated and characterized on A. hydrophila strain TF7. Spontaneous mutants resistant to PM3 were non-motile having lost their characteristic monopolar flagellum. In addition, purified flagella inactivated PM3. PM3 is the first filamentous bacteriophage isolated on Aeromonas, the adsorption site for which is the monopolar flagellum.     

Degradation of benzyldimethylalkylammonium chloride by Aeromonas hydrophila sp. K

AIMS: The aim of this work was to study the biodegradation of benzyldimethylalkylammonium chloride (BAC) by Aeromonas hydrophila sp. K, an organism isolated from polluted soil and capable of utilizing BAC as sole source of carbon and energy. METHODS AND RESULTS: High performance liquid chromatography and gas chromatography-mass spectrometry (GC-MS) analysis was used to study BAC degradation pathway. It was shown that during BAC biodegradation, formation of benzyldimethylamine, benzylmethylamine, benzylamine, benzaldehyde and benzoic acid occurred. Formation of benzyldimethylamine as the initial metabolite suggested that the cleavage of Calkyl-N bond occurred as the first step of BAC catabolism. Liberation of benzylmethylamine and benzylamine likely resulted from subsequent demethylation reactions, followed by deamination with formation of benzaldehyde. Benzaldehyde was rapidly converted into benzoic acid, which was further degraded. CONCLUSIONS: Aer. hydrophila sp. K is able to degrade BAC. A degradation pathway for BAC and related compounds is proposed. SIGNIFICANCE AND IMPACT OF STUDY: These findings are significant for understanding biodegradation pathways of benzyl-containing quaternary ammonium compounds.     

Cloning and characterization of an extracellular temperature-labile serine protease gene from Aeromonas hydrophila

Aeromonas virulence is thought to depend on multigenic functions. The gene for an extracellular protease from Aeromonas hydrophila SO2/2 was cloned in Escherichia coli C600-1 by using pIJ860, bifunctional plasmid, as a vector. The gene encodes for a temperature-labile serine protease (P2) with a molecular mass of approx. 68 kDa which is highly inhibited by PMSF. The gene was expressed in Streptomyces lividans 1326 by transforming protoplasts with the original clone pPA2. We were also able to transfer and express the prt P2 gene in Pseudomonas putida by mating experiments. The protein P2 was secreted into the periplasms of both P. putida and E. coli C600-1 being identical in properties to one of the proteases secreted into the culture supernatant by A. hydrophila SO2/2.     

嗜水气单胞菌国内分离株的毒力因子分布与致病性相关性分析

为了研究分析国内致病性嗜水气单胞菌的毒力因子的分布及与致病性的相关性及其防治,选取5种主要毒力因子气溶素(aerA)、溶血素(hlyA)、丝氨酸蛋白酶(ahpA)、抗金属蛋白酶(ast)、肠毒素(altA)设计5对特异性引物,用PCR方法进行检测,采用急性毒性实验方法测定菌株对异育银鲫的半数致死量(50%lethal dose,LD50),细菌药物敏感试验纸片扩散(K-B)法检测对抗生素的敏感性,结果显示:除菌株Hong12的LD50约为109 cfu,毒力弱外,其他毒力均较强,LD50均在106 cfu左右。aerA、ahpA、hlyA、altA、ast 5种毒力基因的携带率分别为77.8%、88.9%、100%、100%、77.8%。通过比较嗜水气单胞菌毒力基因的分布情况与其对鲫鱼致病性试验结果可以发现,aerA、ahpA为嗜水气单胞菌致病的主要毒力因子,与致病性存在相关性,ast与致病力存在一定相关性,但不是主要毒力因子,hlyA、altA与致病性不存在相关性。9株嗜水气单胞菌对青霉素、羧苄青霉素、万古霉素不敏感,对复合磺胺、头孢噻肟、利福平中毒敏感,对呋喃妥因、氯霉素、四环素、氧氟沙星高度敏感。     

细胞壁缺陷白喉棒状杆菌致病作用的观察及其毒力基因检测

目的比较白喉棒状杆菌亲代细菌型及其稳定L型动物致病性的差异,了解白喉棒状杆菌稳定L型变异的特点,探讨其变异的性质及其与细胞壁缺陷突变的关系。方法用氨苄青霉素在非高渗培养基内人工诱导产毒性白喉棒状杆菌为稳定L型。采用白喉棒状杆菌稳定L型纯培养物及其代谢产物皮内感染家兔,观察局部感染部位皮肤或全身的病理改变。采用微量法提取白喉棒状杆菌稳定L型的染色体DNA,用Tox基因特异性引物进行PCR扩增以检测毒素蛋白结构基因,并进行序列测定和分析。结果白喉棒状杆菌在氨苄青霉素作用下可发生细胞壁缺陷而成为L型,白喉棒状杆菌稳定L型不能引起动物局部或全身发生异常表现,该稳定L型的传代培养物可仍然保留同其亲代细菌型一致的Tox基因及其核苷酸序列。结论提示细胞壁缺失将导致白喉棒状杆菌与产生毒素蛋白有关结构基因在宿主菌细胞内的表达受到抑制,以致使白喉棒状杆菌稳定L型丧失了产生外毒素致病的作用。     

Evaluation of an in vitro cell assay to select attenuated bacterial mutants of Aeromonas hydrophila and Edwardsiella tarda to channel catfish

Aims: To evaluate the feasibility of using an in vitro cell assay to select attenuated bacterial mutants. Methods and Results: Using catfish gill cells G1B, the feasibility of using an in vitro assay instead of in vivo virulence assay using live fish to select attenuated bacterial mutants was evaluated in this study. Pearson correlation analysis between in vitro virulence to G1B cells and in vivo virulence of Aeromonas hydrophila and Edwardsiella tarda revealed that there was a significant correlation between the two (r = ?0·768, P value = 3·7 × 10^?16). Conclusions: The in vitro cell assay might be initially used to screen large quantities of bacteria to select attenuated mutants of catfish pathogens. Significance and Impact of the Study: The in vitro cell assay using catfish gill cells to identify attenuated mutants of catfish pathogens will reduce cost involved in the in vivo virulence assay that requires many fish and aquariums.     

Factors affecting preference responses of the freshwater ciliate Uronema nigricans to bacterial prey

ABSTRACT. To enhance our understanding of the factors affecting feeding selectivity of bacterivorous protists in aquatic systems, we examined the preference responses of the freshwater ciliate Uronema nigricans towards three bacterial prey taxa, Pseudomonas luteola, Serratia rubidaea , and Aeromonas hydrophila . Potential factors influencing the predator–prey contact rate included the previous feeding history of the ciliate and physiological state of bacteria. Preference indexes were obtained from multiple-choice mazes in which ciliates moved preferentially towards alternative bacteria or the prey species on which they had been feeding. Uronema nigricans showed differential attraction towards the offered prey types, and these preferences varied as a function of the ciliate feeding history: U. nigricans growing on P. luteola showed lower preference responses towards the offered bacteria than U. nigricans growing on S. rubidaea . The bacteria in stationary phase elicited a higher degree of attraction than bacteria in exponential phase, probably due to a higher concentration of carbohydrates in the former. Therefore, this protist will preferentially swim towards bacteria in stationary growth phase, although the degree of this response will be affected by the recent feeding history of the ciliate.     

2005～2008年儿童菌痢病原菌与药敏分析及临床意义

目的了解儿童细菌性痢疾病原菌的分布特征及药敏特点,为临床更严谨更规范使用抗生素提供支持与依据。方法对2005年10月至2008年10月57例儿童菌痢的菌型、药敏及耐药性进行分析。结果儿童细菌性痢疾病原菌亚型分类中宋氏痢疾杆菌（D群）占14．0％,福氏痢疾杆菌（B群）占86．0％;痢疾杆菌对常用抗生素耐药率由低到高依次为头孢噻肟,丁胺卡那霉素,庆大霉素,头孢哌酮,头孢三嗪,头孢他啶,头孢唑啉,环丙沙星,氯霉素,复方新诺明,氨苄青霉素;痢疾杆菌单株对多种抗生素的总耐药率为39．2％,多重耐药率为43,9％,且各组间差异无显著性（χ^2=1．608,P=0．996）,痢疾杆菌的耐药问题依然严重。结论新乡市区儿童细菌性痢疾病原菌亚型分类D组已呈明显上升趋势,但总体仍以B群感染为主（86％）;痢疾杆菌的耐药问题依然严重;对儿童菌痢选用抗生素应结合药敏首选头孢噻肟等第三代头孢类抗生素或头孢唑啉,年长儿也可选用丁胺卡那霉素、庆大霉素等氨基苷类,而以环丙沙星为代表的喹诺酮类药物不宜作为儿童尤其7岁以下小儿菌痢的备选药物;氯霉素、复方新诺明及氨苄青霉素已不再作为小儿菌痢的抗菌选择。