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Microbes Inside WM De Vos Wageningen University/Helsinki Antonie van Leeuwenhoek (2009) 95:17–133 DOI 10.1007/s10482-009-9328-5 Abstracts Ó Springer Science+Business Media B.V. 2009 O001 The transmission pathways of parasites, such as anisakid nematodes with indirect life-cycles, are fully included in food webs of aquatic ecosystems. The transmission routes of Microbes inside these nematodes follow closely the trophic relationships W.M. de Vos among their successive hosts, and thus they are parasites embedded in food webs. As a consequence, the completion Wageningen University/Helsinki University of their life-cycles requires a stable trophic web. As a result, Exposure to harsh surfactants, ruthless proteases and pH values the life-cycle of anisakid nematodes in marine ecosystems as low as 1, are extreme conditions that are endured by the with various degrees of habitat disturbance could be affected microbes colonizing the largest microbial ecosystem that is by changes in host population size. Indeed, when the closest to our heart: our microbes inside. Since birth, these population size of the hosts participating in the life-cycle intestinal microbes dominate our body and outnumber our own of these parasites is reduced due to different causes cells by one or more orders of magnitude. Hence, the collective (pollution, by-catch of marine mammals, viral diseases of genome of these microbes, also know as the microbiome, marine mammals, overfishing, etc.), the population size of contributes considerably to the coding capacity of our system. their anisakid endoparasites could also be reduced. This However, unlike our own genome, the microbiome is not, or not would result in a higher probability of genetic drift in the only, vertically inherited and moreover, this personalized organ parasite gene pools and, consequently, a decrease in their can be modified by diet, life style and antimicrobials. Hence, genetic diversity values. It has recently been shown that there is great interest in relating the intestinal microbiome to the distribution of the genetic variability of anisakid health and disease. This requires a quantitative description of nematode populations belonging to several species of the main microbial community members, their genomes and Anisakis, Pseudoterranova and Contracaecum from different functions. Moreover, as the intestinal microbes have developed hosts, in geographical areas with various levels of environ- intimate relations with the host, their dynamics and interactions mental stress in the Boreal and Austral regions, is likely to should be analyzed. This contribution aims to summarize the reflect the influence of a range of factors that could promote recent state of the art in this area with specific attention for their genetic diversity. Austral populations of species describing the microbial diversity in time and space, studying belonging to these three genera exhibited significantly higher the microbe by functional metagenomics, and understanding the genetic variability values than those from the Boreal regions interaction of intestinal bacteria with the host. at both allozyme and mitochondrial gene level. A more remarkable difference in their genetic variability values was observed when only Antarctic and sub-Antarctic popula- tions were compared directly with Arctic and sub-Arctic O002 populations The data are consistent with biotic factors, such as host density of those suitable definitive and intermediate Genetic diversity of nematodes in Arctic and Antarctic hosts for the anisakid nematodes in the Antarctic and sub- ecosystems Antarctic areas, maintaining the high genetic diversity in the anisakid gene pools. Such difference can be explained by the S. Mattiucci lower habitat disturbance of the Antarctic region, which Department of Public Health Sciences, Section of permits the maintenance of more stable trophic webs in this Parasitology, ‘‘Sapienza-University of Rome’’, Rome, Italy ecosystem. 123 18 Antonie van Leeuwenhoek (2009) 95:17–133 O004 O005 Fungal virulence, vertebrate endothermy, and dinosaur Antimicrobial susceptibility testing through EUCAST extinction: is there a connection? G. Kahlmeter A. Casadevall European Committee on Antimicrobial Susceptibility Albert Einstein College of Medicine, Bronx, NY USA Testing (EUCAST) A fundamental question in microbial pathogenesis is the In order to categorise a bacterium as sensitive to an antibiotic, origin of virulence in pathogenic microbes. Virulence is a it is necessary to perform a phenotypic antimicrobial suscep- microbial property that is peculiar in that it is expressed only tibility test. To categorise it as resistant, it may be sufficient to when the microbe encounters a susceptible host. When detect a specific resistance gene. Phenotypic antimicrobial considering the enormous diversity of host-microbial interac- susceptibility testing (AST) is based on the determination of tions that result in the disease state it is insightful to divide the minimum inhibitory concentration (MIC, mg/L) or a pathogenic microbes by whether they were acquired from surrogate of the MIC (inhibition zone diameter in an agar other hosts or directly from the environment. Many microbes diffusion system) and the use of breakpoints. The breakpoints acquired from other hosts usually cause disease only in situ- will determine the categorisation of the bacterium to the ations where there is an alteration in the host-microbe antibiotic as S (susceptible), I (intermediate) or resistant (R) to relationship. However, the origin of virulence for microbes provide information on the likelihood of clinical successshould acquired directly from the environment is less clear since the drug be used for treatment. Breakpoints may be expressed most of these organisms have no need for animal habitation as S B X and R [ Y, as in the EUCAST, or S B X and R C Y, in their life cycle. When considering these questions the as in the CLSI, where X and Y are concentrations correspond- pathogenic fungi, as a group can provide major insights that ing to MIC-values. may be applicable to other organisms. Clinical breakpoints are best determined by breakpoint The most common human pathogenic fungi are Candida committees consisting of experts in the fields of clinical spp. and the causative agents of superficial mycoses. Both these microbiology and infectious diseases. Both CLSI (formerly groups of organisms are acquired from other hosts and disease NCCLS in the USA) and EUCAST (the European Committee almost always follows a disruption of the host-microbe on Antimicrobial Susceptibility Testing) are well known relationship that results in fungal proliferation and damage. breakpoint committees. The former is a commercial enterprise In contrast, most of the other systemic fungal diseases such as engaging the pharmaceutical industry and the profession Aspergillosis, Histoplasmosis, Cryptococcosis, Blastomycoses, whereas the EUCAST is organised by the European Centre etc. are caused by organisms that are acquired directly from the for Disease Control and the European Society for Clinical environment and cause disease usually in situations of immune Microbiology and Infectious Diseases and in close coopera- impairment or infection with large innocula. Studies with C. tion with European Medicines Evaluation Agency. EUCAST neoformans and other fungi have led to the hypothesis that has harmonised breakpoints for Europe and acts as the EMEA selection pressures in the environment, such as predation by breakpoint committee during the process of approval of new amoebae, led to the emergence of microbial traits that could drugs. also function in virulence for human hosts. With these thoughts EUCAST breakpoints are currently being introduced in and ideas at hand, combined with combined with some findings automated systems for AST. The Phoenix from Becton from the Cretaceous-Tertiary boundary it is possible to imagine Dickinson is already validated with EUCAST breakpoints that fungi could have contributed to past extinction events and the Vitek2 from BioMerieux is to follow. A disk test based including the demise of the dinosaurs. on Mueller Hinton agar and 108 cfu/mL inoculum is currently References being developed by EUCAST. Casadevall A, Pirofski L (2007) Accidental Virulence, cryptic New therapeutic traditions, dosage practices, new tools for pathogenesis, martians, lost hosts, and the pathogenicity of setting breakpoints and most importantly new resistance environmental microbes. Eukaryotic Cell 6:2169–74. mechanisms necessitates an evolutionary process for clinical Casadevall A (2006) The cards of virulence and the global breakpoints. EUCAST has an active process for reviewing and virulome. Microbe1:359–364. revising breakpoints. Casadevall, A. (2005). Fungal virulence, vertebrate endo- EUCAST offers free documents on breakpoints, methods thermy and dinosaur extinction: is there a connection? and interpretive criteria on its website (www.eucast.org) which Fungal Genetics and Biol 42:98–106. also displays MIC- (and eventually zone diameter-) distribu- Casadevall A and Pirofski L (2003) The ‘damage-response’ tions of wild type bacteria on the website. These can be used framework of microbial pathogenesis. Nature Microbiology for calibration of methods. Reviews 1:17–24. Steenbergen J, Shuman H, Casadevall A (2001). ‘‘Cryptococ- What is EUCAST’s mission? cus neoformans interactions with amoebae suggest an explanation for its virulence and intracellular pathogenic – To harmonise and regularly review European breakpoints strategy in macrophages. PNAS 98:15245–15250. for phenotypic
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