Investigation of class 1 integrons in Klebsiella pneumoniae clinical and microbiota isolates belonging to different phylogenetic groups in Recife, State of Pernambuco

Introduction The high prevalence of Klebsiella pneumoniae infections is related to the ability of K. pneumoniae to acquire and disseminate exogenous genes associated with mobile elements, such as R plasmids, transposons and integrons. This study investigated the presence of class 1 integrons in clinical and microbiota isolates of K. pneumoniae belonging to different phylogenetic groups and correlated these results with the antimicrobial resistance profiles of the studied isolates. Methods Of the 51 isolates of K. pneumoniae selected for this study, 29 were from multidrug-resistant clinical isolates, and 22 were from children's microbiota. The susceptibility profile was determined using the disk diffusion method, and class 1 integrons were detected through polymerase chain reaction (PCR). Results The results showed that none of the 22 microbiota isolates carried class 1 integrons. Among the 29 clinical isolates, 19 (65.5%) contained class 1 integrons, and resistance to sulfamethoxazole/trimethoprim was identified in 18 of these isolates (94.7%). Among the K. pneumoniae isolates with class 1 integrons, 47% belonged to the KpI phylogenetic group, and one isolate (14.3%) carrying these genetic elements belonged to the KpIII group. Conclusions The wide variety of detected class 1 integrons supports the presence of high rates of antimicrobial resistance, genetic variability, and rapid dissemination of beta-lactamase genes among K. pneumoniae clinical isolates in recent years in hospitals in Recife-PE, Brazil. The findings of this study indicate that the surveillance of K. pneumoniae integrons in clinical isolates could be useful for monitoring the spread of antibiotic resistance genes in the hospital environment.

Evolutionary dynamics of Chlamydia trachomatis genome and identification of molecular patterns of hypothetical protein coding genes

The obligate intracellular bacterium Chlamydia trachomatis is a human pathogen of major public health significance. Strains can be classified into 15 main serovars (A to L3) that preferentially cause ocular infections (A-C), genital infections (D-K) or lymphogranuloma venereum (LGV) (L1-L3), but the molecular basis behind their distinct tropism, ecological success and pathogenicity is not welldefined. Most chlamydial research demands culture in eukaryotic cell lines, but it is not known if stains become laboratory adapted. By essentially using genomics and transcriptomics, we aimed to investigate the evolutionary patterns underlying the adaptation of C. trachomatis to the different human tissues, given emphasis to the identification of molecular patterns of genes encoding hypothetical proteins, and to understand the adaptive process behind the C. trachomatis in vivo to in vitro transition. Our results highlight a positive selection-driven evolution of C. trachomatis towards nichespecific adaptation, essentially targeting host-interacting proteins, namely effectors and inclusion membrane proteins, where some of them also displayed niche-specific expression patterns. We also identified potential "ocular-specific" pseudogenes, and pointed out the major gene targets of adaptive mutations associated with LGV infections. We further observed that the in vivo-derived genetic makeup of C. trachomatis is not significantly compromised by its long-term laboratory propagation. In opposition, its introduction in vitro has the potential to affect the phenotype, likely yielding virulence attenuation. In fact, we observed a "genital-specific" rampant inactivation of the virulence gene CT135, which may impact the interpretation of data derived from studies requiring culture. Globally, the findings presented in this Ph.D. thesis contribute for the understanding of C.trachomatis adaptive evolution and provides new insights into the biological role of C. trachomatishypothetical proteins. They also launch research questions for future functional studies aiming toclarify the determinants of tissue tropism, virulence or pathogenic dissimilarities among C. trachomatisstrains.

Induction and nosocomial dissemination of carbapenem and polymyxin-resistant Klebsiella pneumoniae

INTRODUCTION:Polymyxins are antimicrobial agents capable of controlling carbapenemase-producing Klebsiella pneumoniae infection.METHODS: We report a cluster of four patients colonized or infected by polymyxin-resistant and Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae.RESULTS: Three patients were hospitalized in adjacent wards, and two were admitted to the intensive care unit. The index case maintained prolonged intestinal colonization by KPC-producing K. pneumoniae. Three patients received polymyxin B before the isolation of polymyxin-resistant K. pneumoniae.CONCLUSIONS: Colonization by KPC-producing K. pneumoniae and previous use of polymyxin B may be causally related to the development of polymyxin-resistant microorganisms.

Over expression of AdeABC and AcrAB-TolC efflux systems confers tigecycline resistance in clinical isolates of Acinetobacter baumannii and Klebsiella pneumoniae

Abstract: INTRODUCTION: Due to the wide use of tigecycline in the treatment of severe infections caused by multidrug-resistant (MDR) bacteria, clinical resistance to tigecycline has increased in recent years. Here, we investigated the relationship between tigecycline resistance and the expression of efflux pumps. METHODS: Clinical isolates of Acinetobacter baumannii and Klebsiella pneumoniae were consecutively collected from hospitalized patients in three hospitals. The minimum inhibitory concentration (MIC) of tigecycline was determined using the broth microdilution method. Expression levels of efflux pump genes and regulators were examined by quantitative real-time reverse transcription polymerase chain reaction. The correlations between tigecycline MICs and gene expression levels were analyzed. RESULTS: Overall, 1,026 A. baumannii and 725 K. pneumoniae strains were collected. Most strains were isolated from sputum. The tigecycline resistance rate was 13.4% in A. baumannii isolates and 6.5% in K. pneumoniae isolates. Overexpression of AdeABC and AcrAB-TolC efflux systems was observed found in clinical tigecycline-resistant isolates. The tigecycline MIC had a linear relationship with the adeB expression level in A. baumannii isolates, but not with the acrB expression level in K. pneumoniae isolates. There were significant linear trends in the overexpression of ramA as the tigecycline MIC increased in K. pneumoniae isolates. CONCLUSIONS: Tigecycline resistance in A. baumannii and K. pneumoniae was strongly associated with the overexpression of efflux systems. More studies are needed to elucidate whether there are other regulators that affect the expression of adeB in A. baumannii and how ramA affects the expression of acrB in K. pneumoniae.

Secular trends in Klebsiella pneumoniae isolated in a tertiary-care hospital: increasing prevalence and accelerated decline in antimicrobial susceptibility

Abstract: INTRODUCTION Klebsiella pneumoniae has become an increasingly important etiologic agent of nosocomial infections in recent years. This is mainly due to the expression of virulence factors and development of resistance to several antimicrobial drugs. METHODS This retrospective study examines data obtained from the microbiology laboratory of a Brazilian tertiary-care hospital. To assess temporal trends in prevalence and antimicrobial susceptibility, K. pneumoniae isolates were analyzed from 2000 to 2013. The relative frequencies of K. pneumoniae isolation were calculated among all Gram-negative bacilli isolated in each period analyzed. Susceptibility tests were performed using automated systems. RESULTS: From 2000-2006, K. pneumonia isolates comprised 10.7% of isolated Gram-negative bacilli (455/4260). From 2007-2013, this percentage was 18.1% (965/5331). Strictly considering isolates from bloodstream infections, the relative annual prevalence of K. pneumoniae increased from 14-17% to 27-32% during the same periods. A progressive decrease in K. pneumoniae susceptibility to all antimicrobial agents assessed was detected. Partial resistance was also observed to antimicrobial drugs that have been used more recently, such as colistin and tigecycline. CONCLUSIONS Our study indicates that K. pneumoniae has become a major pathogen among hospitalized patients and confirms its recent trend of increasing antimicrobial resistance.

Rational manipulation of mRNA folding free energy allows rheostat control of pneumolysin production by Streptococcus pneumoniae

Rational manipulation of mRNA folding free energy allows rheostat control of pneumolysin production by Streptococcus pneumoniae

MMP-9/RECK imbalance: a mechanism associated with high-grade cervical lesions and genital infection by Human Papillomavirus (HPV) and Chlamydia trachomatis

"Manuscript"

Pneumomediastino espontâneo como apresentação de infecção por Mycoplasma pneumoniae

Introdução: O pneumomediastino espontâneo (PE) define-se pela presença de ar livre no mediastino, sem causa traumática. É raro em idade pediátrica e exige elevada suspeição diagnóstica. Caso Clínico: Adolescente de 17 anos, longilíneo, fumador ocasional, sem história traumática nem patologia pulmonar, observado na Urgência por dor retrosternal intensa acompanhada por dispneia ligeira. Referência a acessos de tosse seca violenta horas antes. O diagnóstico de PE suspeitou-se pela palpação de enfisema subcutâneo supraclavicular e confirmou-se por telerradiografia do tórax mostrando ar livre mediastínico. A investigação revelou infeção por Mycoplasma pneumoniae pelo que foi medicado com macrólido para além do repouso e analgesia propostos para o PE, evoluindo favoravelmente. Discussão/Conclusôes: O PE é um diagnóstico a não esquecer perante dor torácica aguda no adolescente. A presença de sintomas, por vezes, frustres e a evolução habitualmente benigna contribuem para um provável subdiagnóstico. O tabagismo é um fator predisponente. A associação a infeção por Mycoplasma pneumoniae só raramente foi descrita.

Great amount of C.pneumoniae in ruptured plaque vessel segments at autopsy. A comparative study with stable plaques

A possible relationship between C.pneumoniae (CP) infection, atherosclerosis and acute myocardial infarction is a debated matter. Now we performed the search of CP in histological segments of fatal ruptured plaques and of stable plaques by histochemistry (Macchiavello stain), immunohistochemistry and in situ hybridization techniques. Electron microscopy and confocal laser microscopy techniques were used in two additional cases. The semi-quantitification of CP + cells (0-4+) and quantification of lymphocytes demonstrated greater amount of CP + cells and more inflammation in the adventitia of vulnerable plaque vessel segments than of stable ones, larger amount of CP + cells in adventitia than in the plaque and high frequency of CP + cells in all groups studied. This preliminary study strongly suggests a direct pathogenetic involvement of adventitial CP in the rupture of the atheromatous plaque, development of acute myocardial infarction and also in the development of atherosclerosis.

Estudio de epitopes de antígenos para interpretar su respuesta inmune. Modelo 1: Virus rubeola. Modelo 2: Bacteria Chlamydia trachomatis

El estudio de los epitopes de los antígenos permitirá no sólo la diferenciación de cepas sino también la interpretación correcta de la respuesta inmune. Modelo1: Rubeola es una enfermedad infecciosa caracterizada por una erupción localizada, fiebre y adenopatías, que por lo general se produce en niños de edad escolar o preescolar. El virus de la rubeola es el único miembro del género ribivirus en la familia de los Togavirus. Se sabe que es una partícula esférica que mide 60-90 nm y lleva la información genética en una sola cadena de RNA de polaridad positiva formando una cápside icosahédrica. Estructuralmente está compuesto por tres proteínas, una no glicosilada, asociada al RNA en la nucleocápside, llamada C, y dos glicoproteínas E1 y E2 que se encuentran en la envoltura del virus donde se presentan formando complejos por dímeros E1-E1 y E1-E2. Uno de los motivos por los que se realizan estudios comparativos entre diferentes cepas de virus rubeola radica en la observación de que la respuesta inmune frente a la infección con la cepa salvaje es más eficiente y duradera que la inducida por la cepa vacunal y que no se conocen fehacientemente si la capacidad teratogénica del virus depende de la cepa viral o del tipo de infección que se produce en la placenta y en el feto. La disminución o desaparición de la respuesta inmune específica puede posibilitar la reinfección de una persona vacunada, lo que adquiere particular importancia en el caso de las embarazadas. Es por ello que este estudio se centra en el análisis comparativo de las propiedades biológicas y estructurales de la cepa de virus rubeola de circulación local (cepa Córdoba y sus clones) frente a las cepas Glichrist (prototipo) y RA 27/3 (vacunal). Esta parte del trabajo contribuye al proyecto mundial de obtención de una vacuna sintética o infectiva para controlar la infección por el virus rubeola. Objetivos: 1. Estudiar la cinética de aparición de los epitopes en citoplasma y membrana celular con una técnica de inmunofuorescencia indirecta de células infectadas y bloquear a distintos pasos la vía de síntesis de proteínas en las células infectadas para producir el camino de síntesis del complejo E1-E1 hasta su aparición en membrana. 2. Realizar la técnica de mapeo peptídico para la proteína E2 en diferentes cepas. Modelo 2: Chlamydia trachomatis es una bacteria intracelular obligada de un ciclo de vida dimórfico, con una fase extracelular llamada Cuerpo Elemental (CE), que es la forma infectiva y metabólicamente inactiva y una fase intracelular llamada Cuerpo Reticular que es la forma replicada y metabólicamente activa de la bacteria. Es el agente etiológico de Enfermedades de Transmisión Sexual (ETS) más comunmente aislado en poblaciones de riesgo, además de ser la primera causa de ceguera prevenible a nivel mundial. El resultado de la infección con C. trachomatis puede terminar en inmunidad o enfermedad dependiendo de la interacción del sistema inmune del huésped con los antígenos chlamydiales específicos. El estudio de los antígenos que generan la respuesta inmune humoral como los tipos e isotipos de inmunoglobulinas producidas en esta respuesta, en las poblaciones con diferentes manifestaciones de la infección por C. trachomatis , podrían asociarse a un tipo de perfil de respuesta de linfocitos TH y dar un valor pronóstico de la evolución de la infección. Objetivo: 1. Separar y estudiar algunos de los más importantes antígenos de la bacteria Chlamydia trachomatis .

Biotipagem e resistotipagem para o traçado epidemiológico da origem fecal de Klebsiella pneumoniae em infecções urinárias

Em 24 (82,7%) de 29 pacientes com infecção urinária por Klebsiella pneumoniae isolamos a mesma espécie a partir de amostras fecais. Estudamos 24 cepas urinárias e 219 cepas fecais encontrando 50 biótipos distintos (em média quatro biótipos por amostra fecal). Em dez (34,4%) dos 29 pacientes o biótipo de uma ou mais cepas fecais corresponderam ao biotipo da cepa urinária - sem encontrarmos associação entre a simultaneidade e a prévia cateterização vesical (p>0,05). Na resistotipagem - utilizando quatro substâncias químicas previamente escolhidas entre 34 produtos testados (verde brilhante, verde malaquita, telurito de potássio e cloreto mercúrico) encontramos 16 resistotipos distintos. Em 14 (58,3%), dos 24 casos houve detecção do mesmo resistotipo em cepa(s) fecal(is) e urinária do mesmo paciente, entretanto, só em cinco (20,9%) dos casos houve concordância com a biotipagem na indicação de simultaneidade. A concordância de resultados quanto à ausência ou presença de biotipos ou resistotipos simultâneos, na urina e nas fezes, foi de somente 54,2%. A presença de resistência aos íons telurito e mercúrio, entre cepas fecais e cepas urinárias, no mesmo paciente, estava significativamente associada (p<0,001).

Chlamydia-related bacteria in respiratory samples in Finland.

Chlamydia-related bacteria, new members of the order Chlamydiales, are suggested to be associated with respiratory disease. We used real-time PCR to investigate the prevalence of Parachlamydia acanthamoebae, Protochlamydia spp., Rhabdochlamydia spp., Simkania negevensis and Waddlia chondrophila in samples taken from patients with suspected respiratory tract infections. Of the 531 samples analyzed, the subset of 136 samples contained 16 (11.8%) samples positive for Rhabdochlamydia spp. DNA. P. acanthamoebae, Protochlamydia spp., S. negevensis and W. chondrophila DNA were not detected among the respiratory samples investigated. These results suggest an association of Rhabdochlamydia spp. with respiratory disease.

Macrophage Migration Inhibitory Factor Deficiency Is Associated With Impaired Killing of Gram-Negative Bacteria by Macrophages and Increased Susceptibility to Klebsiella pneumoniae Sepsis.

The cytokine macrophage migration inhibitory factor (MIF) is an important component of the early proinflammatory response of the innate immune system. However, the antimicrobial defense mechanisms mediated by MIF remain fairly mysterious. In the present study, we examined whether MIF controls bacterial uptake and clearance by professional phagocytes, using wild-type and MIF-deficient macrophages. MIF deficiency did not affect bacterial phagocytosis, but it strongly impaired the killing of gram-negative bacteria by macrophages and host defenses against gram-negative bacterial infection, as shown by increased mortality in a Klebsiella pneumonia model. Consistent with MIF's regulatory role of Toll-like 4 expression in macrophages, MIF-deficient cells stimulated with lipopolysaccharide or Escherichia coli exhibited reduced nuclear factor κB activity and tumor necrosis factor (TNF) production. Addition of recombinant MIF or TNF corrected the killing defect of MIF-deficient macrophages. Together, these data show that MIF is a key mediator of host responses against gram-negative bacteria, acting in part via a modulation of bacterial killing by macrophages.

Role of NOX enzymes in phagocyte response and signaling upon Chlamydia infection

Les membres de l'ordre des Chlamydiales peuvent infecter un choix étendu d'animaux, insectes, et protistes. Comme toutes bactéries intracellulaires obligatoires, les Chlamydiales ont besoin d'une cellule hôte pour se répliquer. Chaque fois qu'une cellule est infectée une lutte commence entre les mécanismes de défense de la cellule et l'arsenal de facteurs de virulence de la bactérie. Dans cette thèse nous nous sommes intéressés à déterminer le rôle de deux mécanismes de l'immunité innée de l'hôte. En premier, nous avons étudié les NADPH oxidases, une source de molécules superoxydantes (MSO). Leur rôle dans la restriction de la réplication de Waddlia chondrophila et Estrella iausannensis a été étudié dans l'organisme modèle Dictyostelium discoideum et les macrophages humains. Différentes protéines Nox étaient nécessaires pour contrôler la réplication de W. chondrophila ou E. Iausannensis. De plus, nous avons déterminé que parmi les Chlamydiales, cinq espèces possédaient une catalase. Cette enzyme peut dégrader l'eau oxygénée, une MSO. L'activité de la catalase a été démontrée in vitro et dans les corps élémentaires. Avant de pouvoir étudier le rôle de NOX2 dans des macrophages infectés avec E. Iausannensis, nous avons dû établir la capacité de la bactérie à se répliquer clans les macrophages avec son trafic intracellulaire. Le deuxième mécanisme d'immunité innée que nous avons étudié est l'autophagie. Dans les cellules infectées l'autophagie permet de digérer les bactéries envahissantes. Deux protéines de la voie autophagique (Atg1 et Atg8) jouent un rôle dans la restriction de la croissance de W. chondrophila dans D. discoideum. D'avantage d'études sur l'immunité innée et les bactéries apparentés aux Chlamydia sont indispensables, car les réponses paraissent être spécifiques pour chaque espèce. - Members of the Chlamydiales order are able to infect a large variety of animals, insects, and protists. These obligate intracellular bacteria require a host cell for replication. Each time a cell is infected a struggle begins between the virulence arsenal of the bacteria and the defense mechanisms activated by the host. Each bacterial species will exhibit a selection of virulence factors that will allow it to overcome the defense of the host in some species, but not others. In this thesis we were interested in dissecting the role of two host innate immunity mechanisms. First we determined the role of NADPH oxidases, a source of reactive oxygen species (ROS), in restricting replication of Waddlia chondrophila and EstreHa lausannensis in the model organism Dictyostelium discoideum and human macrophages. Different Nox proteins were required to restrict growth of W. chondrophila and E. lausannensis. Additionally, we determined that five Chlamydia- related bacterial species encode for catalase, an enzyme that is able to degrade hydrogen peroxide, a ROS. The activity of the catalase was demonstrated in vitro and in elementary bodies. To study the role of NOX2 in macrophages for E. lausannensis we first had to determine the ability of E. lausannensis to grow in macrophages. Besides demonstrating its replication we also determined the intracellular trafficking of E. lausannensis. The second innate immunity mechanism studied was autophagy. Through autophagy bacteria can be targeted to degradation. Atg1 and Atg8, two autophagic proteins appeared restrict W. chondrophila replication in D. discoideum. More studies on innate immunity and Chlamydia-related bacteria are required. It appears that the responses to innate immunity are species specific and it will be difficult to generalize data obtained for W. chondrophila to the Chlamydiales order.

Genomics of "chlamydia"-related bacteria

AbstractThe Chlamydiales order is an important bacterial phylum that comprises some of the most successful human pathogens such as Chlamydia trachomatis, the leading infectious cause of blindness worldwide. Since some years, several new bacteria related to Chlamydia have been discovered in clinical or environmental samples and might represent emerging pathogens. The genome sequencing of classical Chlamydia has brought invaluable information on these obligate intracellular bacteria otherwise difficult to study due to the lack of tools to perform basic genetic manipulation. The recent emergence of high-throughput sequencing technologies yielding millions of reads in a short time lowered the costs of genome sequencing and thus represented a unique opportunity to study Chlamydia-re\ated bacteria. Based on the sequencing and the analysis of Chlamydiales genomes, this thesis provides significant insights into the genetic determinants of the intracellular lifestyle, the pathogenicity, the metabolism and the evolution of Chlamydia-related bacteria. A first approach showed the efficacy of rapid sequencing coupled to proteomics to identify immunogenic proteins. This method, particularly useful for an emerging pathogen such as Parachlamydia acanthamoebae, enabled us to discover good candidates for the development of diagnostic tools that would permit to evaluate at larger scale the role of this bacterium in disease. Second, the complete genome of Waddlia chondrophila, a potential agent of miscarriage, encodes numerous virulence factors to manipulate its host cell and resist to environmental stresses. The reconstruction of metabolic pathways showed that the bacterium possesses extensive capabilities compared to related organisms. However, it is still incapable of synthesizing some essential components and thus has to import them from its host. Third, the genome comparison of Protochlamydia naegleriophila to its closest known relative Protochlamydia amoebophila revealed a particular evolutionary dynamic with the occurrence of an unexpected genome rearrangement. Fourth, a phylogenetic analysis of P. acanthamoebae and Legionella drancourtii identified several genes probably exchanged by horizontal gene transfer with other intracellular bacteria that might occur within their amoebal host. These genes often encode mechanisms for resistance to metal or toxic compounds. As a whole, the analysis of the different genomes enabled us to highlight a large diversity in size, GC percentage, repeat content as well as plasmid organization. The abundant genomic data obtained during this thesis have a wide impact since they provide the necessary bases for detailed investigations on countless aspects of the biology and the evolution of Chlamydia-related bacteria, whether in wet lab or by bioinformatical analyses.RésuméL'ordre des Chlamydiales est un important phylum bactérien qui comprend de nombreuses espèces pathogènes pour l'homme et les animaux, dont Chlamydia trachomatis, responsable du trachome, la cause majeure de cécité d'origine infectieuse à travers le monde. Durant ces dernières décennies, de nombreuses bactéries apparentées aux Chlamydia ont été découvertes dans des échantillons environnementaux ou cliniques mais leur éventuel rôle pathogène dans le développement de maladies reste peu connu. Ces bactéries sont des intracellulaires obligatoires car elles ont besoin d'une cellule hôte pour se multiplier, ce qui rend leur étude particulièrement difficile. Le développement de nouvelles technologies permettant de séquencer le génome d'un organisme rapidement et à moindre coût ainsi que l'essor des méthodes d'analyse s'y rapportant représentent une opportunité exceptionnelle d'étudier ces organismes. Dans ce contexte, cette thèse démontre l'utilité de la génomique pour développer de nouveaux outils diagnostiques ainsi que pour étudier le métabolisme de ces bactéries, leurs facteurs de virulence et leur évolution.Ainsi, une première approche a illustré l'utilité d'un séquençage rapide pour obtenir les informations nécessaires à l'identification de protéines qui sont reconnues par des anticorps humains ou animaux. Cette méthode, particulièrement utile pour un pathogène émergent tel que Parachlamydia acanthamoebae, a permis de découvrir de bons candidats pour le développement d'un outil diagnostique qui permettrait d'évaluer à plus large échelle le rôle de cette bactérie notamment dans la pneumonie. L'analyse du contenu génique de Waddlia chondrophila, un autre germe qui pourrait être impliqué dans les avortements et tes fausses-couches, a en outre mis en évidence la présence de nombreux facteurs connus qui lui permettent de manipuler son hôte. Cette bactérie possède de plus grandes capacités métaboliques que les autres Chlamydia, mais elle est incapable de synthétiser certains composants et doit donc les importer de son hôte pour subvenir à ses besoins. La comparaison du génome de Protochlamydia naegleriophila à son plus proche parent, Protochlamydia amoebophila, a dévoilé une évolution dynamique particulière avec l'occurrence d'un réarrangement majeur inattendu après la séparation de ces deux espèces. En outre, ces études ont montré l'occurrence de plusieurs transferts de gène avec d'autres organismes plus éloignés, notamment d'autres intracellulaires d'amibes, souvent pour l'acquisition de mécanismes de résistances à des composés toxiques. Les données génomiques acquises durant ce travail posent les fondements nécessaires a de nombreuses analyses qui permettront progressivement de mieux comprendre de nombreux aspects de ces bactéries fascinantes.