鲟源豚鼠气单胞菌分离与强弱毒株病原生物学比较研究

从患慢性出血败血综合症的杂交鲟和达氏鳇病鱼标本中分离到一株优势菌,优势率80%,经回感和重分离确定为致病菌,并经生理生化和16SrDNA检测为豚鼠气单胞菌,命名为ZZF071029（简称ZF1029）,以豚鼠气单胞菌弱毒株ZF1029和强毒株XL2-T为研究对象,进行了菌落形态学、运动性实验、溶血性实验、小鼠致死试验、角膜毒力实验、药物敏感性实验等病原生物学相关实验。实验结果证明,病原菌运动性、溶血性、小鼠致死力、角膜毒力与菌株毒力和致病力正相关,并与所致杂交鲟和达氏鳇出血败血综合症急、慢性程度和致死率一致。     

A draft genome of Yersinia pestis from victims of the Black Death

Technological advances in DNA recovery and sequencing have drastically expanded the scope of genetic analyses of ancient specimens to the extent that full genomic investigations are now feasible and are quickly becoming standard. This trend has important implications for infectious disease research because genomic data from ancient microbes may help to elucidate mechanisms of pathogen evolution and adaptation for emerging and re-emerging infections. Here we report a reconstructed ancient genome of Yersinia pestis at 30-fold average coverage from Black Death victims securely dated to episodes of pestilence-associated mortality in London, England, 1348-1350. Genetic architecture and phylogenetic analysis indicate that the ancient organism is ancestral to most extant strains and sits very close to the ancestral node of all Y. pestis commonly associated with human infection. Temporal estimates suggest that the Black Death of 1347-1351 was the main historical event responsible for the introduction and widespread dissemination of the ancestor to all currently circulating Y. pestis strains pathogenic to humans, and further indicates that contemporary Y. pestis epidemics have their origins in the medieval era. Comparisons against modern genomes reveal no unique derived positions in the medieval organism, indicating that the perceived increased virulence of the disease during the Black Death may not have been due to bacterial phenotype. These findings support the notion that factors other than microbial genetics, such as environment, vector dynamics and host susceptibility, should be at the forefront of epidemiological discussions regarding emerging Y. pestis infections.     

The role of evolution in the emergence of infectious diseases

It is unclear when, where and how novel pathogens such as human immunodeficiency virus (HIV), monkeypox and severe acute respiratory syndrome (SARS) will cross the barriers that separate their natural reservoirs from human populations and ignite the epidemic spread of novel infectious diseases. New pathogens are believed to emerge from animal reservoirs when ecological changes increase the pathogen's opportunities to enter the human population and to generate subsequent human-to-human transmission. Effective human-to-human transmission requires that the pathogen's basic reproductive number, R(0), should exceed one, where R(0) is the average number of secondary infections arising from one infected individual in a completely susceptible population. However, an increase in R(0), even when insufficient to generate an epidemic, nonetheless increases the number of subsequently infected individuals. Here we show that, as a consequence of this, the probability of pathogen evolution to R(0) > 1 and subsequent disease emergence can increase markedly.     

鸡大肠杆菌病致病因素及其研究进展

对鸡大肠杆菌致病力或毒力因子等相关致病因素作了简要概述,讨论了相关领域的研究进展.1）大肠杆菌的菌毛抗原是一种特异蛋白质,可以被免疫系统所识别,因此在疾病的免疫预防和相关疫苗研制等方面有着重要意义.2）鸡大肠杆菌肠毒素中LTB和STn亚单位具有良好的免疫原性,认为在降低ST的生物毒性有所突破的前提下,借用基因重组融合技术以及优化重组基因表达,是利用该毒素因子免疫防治的发展趋势.3）诸多实验研究表明,提纯的大肠杆菌OMP或通过基因工程表达的OMP同样具有良好的免疫原性和保护作用.某些细菌的不同血清型间有相同的OMP免疫原,这为OMP亚单位疫苗以至于基因疫苗的研发都提供了理论基础.     

The plant immune system

Many plant-associated microbes are pathogens that impair plant growth and reproduction. Plants respond to infection using a two-branched innate immune system. The first branch recognizes and responds to molecules common to many classes of microbes, including non-pathogens. The second responds to pathogen virulence factors, either directly or through their effects on host targets. These plant immune systems, and the pathogen molecules to which they respond, provide extraordinary insights into molecular recognition, cell biology and evolution across biological kingdoms. A detailed understanding of plant immune function will underpin crop improvement for food, fibre and biofuels production.     

Widespread distribution and fitness contribution of Xanthomonas campestris avirulence gene avrBs2

Disease-resistance genes introduced into cultivated plants are often rendered ineffective by the ability of pathogen populations to overcome host resistance. The bacterial pathogen Xanthomonas campestris pathovar vesicatoria causes bacterial spot disease of tomato and pepper, and this pathogen has been shown to overcome disease resistance in pepper (Capsicum annuum) by evading the recognition and defence response of the host plant. Numerous resistance genes to bacterial spot have been identified in pepper and its wild relatives, each providing resistance to specific races of X.c. vesicatoria. The resistance gene Bs1, for example, provides resistance to X.c. vesicatoria strains expressing the avirulence gene avrBs1; Bs2 provides resistance to stains expressing avrBs2 and so on. We now report that avr Bs2 is highly conserved among strains of X.c. vesicatoria, and among many other pathovars of X. campestris. Furthermore, we find that avrBs2 is in fact needed for full virulence of the pathogen on susceptible hosts. This implies that plants carrying Bs2 can recognize an essential gene of the bacterial pathogen, which may explain why Bs2 confers the only effective field resistance to X.c. vesicatoria in pepper.     

Recovery of immunodeficient mice from a vaccinia virus/IL-2 recombinant infection

Vaccinia virus recombinants that express cloned genes encoding antigens of unrelated infectious agents, such as hepatitis B virus and human immunodeficiency virus (HIV), provide a new approach to the development of live vaccines. Although there is evidence that genetically engineered vaccinia viruses have reduced pathogenicity a major obstacle to their use as vaccines is that severe complications can occur after vaccination, especially in immunodeficient individuals. We describe here a recombinant vaccinia virus expressing murine interleukin-2 (IL-2) and show that athymic nude mice infected with the recombinant virus resolve the virus infection rapidly whereas mice infected with control virus develop a progressive vaccinal disease. By incorporating the gene for IL-2 in live virus vaccines it may be possible to prevent the severe complications that arise in recipients with an impaired immune system.     

Detection of prokaryotic mRNA signifies microbial viability and promotes immunity

Live vaccines have long been known to trigger far more vigorous immune responses than their killed counterparts. This has been attributed to the ability of live microorganisms to replicate and express specialized virulence factors that facilitate invasion and infection of their hosts. However, protective immunization can often be achieved with a single injection of live, but not dead, attenuated microorganisms stripped of their virulence factors. Pathogen-associated molecular patterns (PAMPs), which are detected by the immune system, are present in both live and killed vaccines, indicating that certain poorly characterized aspects of live microorganisms, not incorporated in dead vaccines, are particularly effective at inducing protective immunity. Here we show that the mammalian innate immune system can directly sense microbial viability through detection of a special class of viability-associated PAMPs (vita-PAMPs). We identify prokaryotic messenger RNA as a vita-PAMP present only in viable bacteria, the recognition of which elicits a unique innate response and a robust adaptive antibody response. Notably, the innate response evoked by viability and prokaryotic mRNA was thus far considered to be reserved for pathogenic bacteria, but we show that even non-pathogenic bacteria in sterile tissues can trigger similar responses, provided that they are alive. Thus, the immune system actively gauges the infectious risk by searching PAMPs for signatures of microbial life and thus infectivity. Detection of vita-PAMPs triggers a state of alert not warranted for dead bacteria. Vaccine formulations that incorporate vita-PAMPs could thus combine the superior protection of live vaccines with the safety of dead vaccines.     

闽南地区紫石斑(Epinephelus lanceolatus)幼鱼爆发性传染病的病原学研究

应用RT-PCR技术．检测了福建南部石斑鱼育苗场患病紫石斑鱼的病原．结果检出神经坏死病毒；对该病毒的RT-PCR产物进行了核酸测序和序列分析．与基因库中马拉巴斑和紫石斑神经坏死病毒编码病毒核衣壳蛋白的基因片段序列的同源性大于97％．病原菌分离鉴定结果．有2菌株分别是溶藻弧菌和豚鼠气单胞菌．对实验动物有一定的毒力．药敏试验表明这些菌株对常用抗生素表现耐药．临床症状和实验结果都表明．神经坏死病毒感染是导致石斑鱼大批死亡的主要原因．而溶藻弧菌和豚鼠气单胞菌应为继发感染．加重了病情．使更多的石斑鱼死亡．     

Antiviral treatment is more effective than smallpox vaccination upon lethal monkeypox virus infection

There is concern that variola virus, the aetiological agent of smallpox, may be used as a biological weapon. For this reason several countries are now stockpiling (vaccinia virus-based) smallpox vaccine. Although the preventive use of smallpox vaccination has been well documented, little is known about its efficacy when used after exposure to the virus. Here we compare the effectiveness of (1) post-exposure smallpox vaccination and (2) antiviral treatment with either cidofovir (also called HPMPC or Vistide) or with a related acyclic nucleoside phosphonate analogue (HPMPO-DAPy) after lethal intratracheal infection of cynomolgus monkeys (Macaca fascicularis) with monkeypox virus (MPXV). MPXV causes a disease similar to human smallpox and this animal model can be used to measure differences in the protective efficacies of classical and new-generation candidate smallpox vaccines. We show that initiation of antiviral treatment 24 h after lethal intratracheal MPXV infection, using either of the antiviral agents and applying various systemic treatment regimens, resulted in significantly reduced mortality and reduced numbers of cutaneous monkeypox lesions. In contrast, when monkeys were vaccinated 24 h after MPXV infection, using a standard human dose of a currently recommended smallpox vaccine (Elstree-RIVM), no significant reduction in mortality was observed. When antiviral therapy was terminated 13 days after infection, all surviving animals had virus-specific serum antibodies and antiviral T lymphocytes. These data show that adequate preparedness for a biological threat involving smallpox should include the possibility of treating exposed individuals with antiviral compounds such as cidofovir or other selective anti-poxvirus drugs.     

Inflammation and therapeutic vaccination in CNS diseases

The spectrum of inflammatory diseases of the central nervous system has been steadily expanding from classical autoimmune disorders such as multiple sclerosis to far more diverse diseases. Evidence now suggests that syndromes such as Alzheimer's disease and stroke have important inflammatory and immune components and may be amenable to treatment by anti-inflammatory and immunotherapeutic approaches. The notion of 'vaccinating' individuals against a neurodegenerative disorder such as Alzheimer's disease is a marked departure from classical thinking about mechanism and treatment, and yet therapeutic vaccines for both Alzheimer's disease and multiple sclerosis have been validated in animal models and are in the clinic. Such approaches, however, have the potential to induce unwanted inflammatory responses as well as to provide benefit.     

恒河猴和食蟹猴在结核病研究中的应用进展

摘要: 结核病是由结核分枝杆菌引起的慢性消耗性传染病,位居单一病原体引起死亡的严重传染病之首。结核病 历史悠久,对其记录可以追溯到远古社会。人类同结核病进行了数个世纪艰苦卓绝的斗争,然而,时至今日,结核 病仍是严重危害公众健康的全球性公共卫生问题。因此,研发有效的抗结核疫苗和药物迫在眉睫,而开展这些工 作需要合适的实验动物模型。由于非人灵长类动物在解剖结构、生理生化、免疫和代谢等生物学特性方面与人类 高度相似,应用非人灵长类实验动物对结核病的发病机制、机体免疫反应及相关疫苗和药物的深入研究至关重要, 甚至是不可替代的。本文着重就恒河猴和食蟹猴在人类结核病防治方面的研究进展进行综述。     

Herd immunity to helminth infection and implications for parasite control

Despite much research on immunological responses to helminth parasites, knowledge of the dynamic interplay between levels of herd immunity in humans and the rates of exposure, establishment and mortality of parasites remains limited. We describe here a simple mathematical model for the population dynamics of helminth infections which mirrors the development of a degree of acquired immunity within populations which are genetically heterogeneous with respect to immunological responsiveness. We interpret observed patterns in the age-specific intensity of infection and attempt to understand the possible effects of control measures based on chemotherapy and vaccination. Mass chemotherapy can, in some circumstances, reduce the level of herd immunity such that average worm burdens in the adult age classes rise above their precontrol levels. When certain individuals or groups are predisposed to heavy infection, selective or targeted drug treatment can have significantly greater impact than mass or random application. Conversely, model predictions suggest that effective parasite control by vaccination (if and when vaccines become available) is difficult to achieve in communities that are genetically heterogeneous in their ability to mount protective responses to infection.     

The evolution of inaccurate mimics

Instances of strikingly accurate Batesian mimicry (in which a palatable prey organism closely resembles an aversive model) are often cited to illustrate the power of natural selection. Less attention has been paid to those mimics, such as many hoverfly (Syrphidae) mimics of wasps or bees, that resemble their models only poorly. Attempts to provide an adaptive explanation for imperfect mimicry have suggested that what seems a crude resemblance to human observers may appear a close match to predators, or that inaccurate mimics may bear a general resemblance to several different model species. I show here, however, that truly inaccurate mimicry of a single model organism may be favoured over perfect resemblance, by kin selection. Signal detection theory predicts that predators will modify their level of discrimination adaptively in response to the relative frequencies and similarity of models and mimics. If models are rare and/or weakly aversive, greater local similarity of mimics can thus lead to greater attack rates. Where individual mimics are related to others in their vicinity, kin selection will then oppose the evolution of accurate mimicry.     

Large-scale sequencing of human influenza reveals the dynamic nature of viral genome evolution

Influenza viruses are remarkably adept at surviving in the human population over a long timescale. The human influenza A virus continues to thrive even among populations with widespread access to vaccines, and continues to be a major cause of morbidity and mortality. The virus mutates from year to year, making the existing vaccines ineffective on a regular basis, and requiring that new strains be chosen for a new vaccine. Less-frequent major changes, known as antigenic shift, create new strains against which the human population has little protective immunity, thereby causing worldwide pandemics. The most recent pandemics include the 1918 'Spanish' flu, one of the most deadly outbreaks in recorded history, which killed 30-50 million people worldwide, the 1957 'Asian' flu, and the 1968 'Hong Kong' flu. Motivated by the need for a better understanding of influenza evolution, we have developed flexible protocols that make it possible to apply large-scale sequencing techniques to the highly variable influenza genome. Here we report the results of sequencing 209 complete genomes of the human influenza A virus, encompassing a total of 2,821,103 nucleotides. In addition to increasing markedly the number of publicly available, complete influenza virus genomes, we have discovered several anomalies in these first 209 genomes that demonstrate the dynamic nature of influenza transmission and evolution. This new, large-scale sequencing effort promises to provide a more comprehensive picture of the evolution of influenza viruses and of their pattern of transmission through human and animal populations. All data from this project are being deposited, without delay, in public archives.     

Altruism through beard chromodynamics

The evolution of altruism, a behaviour that benefits others at one's own fitness expense, poses a darwinian paradox. The paradox is resolved if many interactions are with related individuals so that the benefits of altruism are reaped by copies of the altruistic gene in other individuals, a mechanism called kin selection. However, recognition of altruists could provide an alternative route towards the evolution of altruism. Arguably the simplest recognition system is a conspicuous, heritable tag, such as a green beard. Despite the fact that such genes have been reported, the 'green beard effect' has often been dismissed because it is unlikely that a single gene can code for altruism and a recognizable tag. Here we model the green beard effect and find that if recognition and altruism are always inherited together, the dynamics are highly unstable, leading to the loss of altruism. In contrast, if the effect is caused by loosely coupled separate genes, altruism is facilitated through beard chromodynamics in which many beard colours co-occur. This allows altruism to persist even in weakly structured populations and implies that the green beard effect, in the form of a fluid association of altruistic traits with a recognition tag, can be much more prevalent than hitherto assumed.     

Parallel evolution of virulence in pathogenic Escherichia coli

The mechanisms underlying the evolution and emergence of new bacterial pathogens are not well understood. To elucidate the evolution of pathogenic Escherichia coli strains, here we sequenced seven housekeeping genes to build a phylogenetic tree and trace the history of the acquisition of virulence genes. Compatibility analysis indicates that more than 70% of the informative sites agree with a single phylogeny, suggesting that recombination has not completely obscured the remnants of ancestral chromosomes. On the basis of the rate of synonymous substitution for E. coli and Salmonella enterica (4.7 x 10(-9) per site per year), the radiation of clones began about 9 million years ago and the highly virulent pathogen responsible for epidemics of food poisoning, E. coli O157:H7, separated from a common ancestor of E. coli K-12 as long as 4.5 million years ago. Phylogenetic analysis reveals that old lineages of E. coli have acquired the same virulence factors in parallel, including a pathogenicity island involved in intestinal adhesion, a plasmid-borne haemolysin, and phage-encoded Shiga toxins. Such parallel evolution indicates that natural selection has favoured an ordered acquisition of genes and the progressive build-up of molecular mechanisms that increase virulence.     

Cooperation and conflict in quorum-sensing bacterial populations

It has been suggested that bacterial cells communicate by releasing and sensing small diffusible signal molecules in a process commonly known as quorum sensing (QS). It is generally assumed that QS is used to coordinate cooperative behaviours at the population level. However, evolutionary theory predicts that individuals who communicate and cooperate can be exploited. Here we examine the social evolution of QS experimentally in the opportunistic pathogen Pseudomonas aeruginosa, and show that although QS can provide a benefit at the group level, exploitative individuals can avoid the cost of producing the QS signal or of performing the cooperative behaviour that is coordinated by QS, and can therefore spread. We also show that a solution to the problem of exploitation is kin selection, if interacting bacterial cells tend to be close relatives. These results show that the problem of exploitation, which has been the focus of considerable attention in animal communication, also arises in bacteria.     

我国新城疫病毒疫苗通过重组影响病毒进化

许多商业新城疫活疫苗被广泛用来阻止我国新城疫病毒的感染.然而,疫苗在我国新城疫病毒群进化过程中所起的作用鲜为人知.为了确定在我国广泛应用的新城疫疫苗是否对该病毒进化进程产生影响,在本研究中,我们对在我国流行的新城疫病毒进行了系统发生和重组分析,获得了一系列NDV株间同源重组的证据.一些重组株的部分遗传物质被发现可能来自疫苗株,这些结果证实疫苗可能通过同源重组影响我国新城疫病毒的进程.由于疫苗与流行株之间的重组可能导致出现疫苗接种失败的后果,因此,在临床上应当尽可能避免疫苗与野生株之间的重组.故在NDV防治过程中,应该采用合适的免疫程序避免这种重组的发生.