Organisms in the genus Kocuria are Gram-positive, coagulase-negative, coccoid actinobacteria belonging to the family Micrococcaceae, suborder Micrococcineae, order Actinomycetales. Sporadic reports in the literature have dealt with infections by Kocuria species, mostly in compromised hosts with serious underlying conditions. Nonetheless, the number of infectious processes caused by such bacteria may be higher than currently believed, given that misidentification by phenotypic assays has presumably affected estimates of the prevalence over the years. As a further cause for concern, guidelines for therapy of illnesses involving Kocuria species are lacking, mostly due to the absence of established criteria for evaluating Kocuria replication or growth inhibition in the presence of antibiotics. Therefore, breakpoints for staphylococci have been widely used throughout the literature to try to understand this pathogen's behaviour under drug exposure; unfortunately, this has sometimes created confusion, thus higlighting the urgent need for specific interpretive criteria, along with a deeper investigation into the resistance determinants within this genus. We therefore review the published data on cultural, genotypic and clinical aspects of the genus Kocuria, aiming to shed some light on these emerging nosocomial pathogens. IntroductionThe genus Kocuria was named after Miroslav Kocur, a Slovakian microbiologist, and belongs to the family Micrococcaceae, suborder Micrococcineae, order Actinomycetales, class Actinobacteria (Takarada et al., 2008;Zhou et al., 2008;Lee et al., 2009;Stackebrandt et al., 1995). It includes Gram-positive, strictly aerobic (a few exceptions are Kocuria kristinae, which is facultatively anaerobic, Kocuria marina, which may grow in 5 % CO 2 , and Kocuria rhizophila strain DC2201, which can proliferate anaerobically), catalasepositive, coagulase-negative, non-haemolytic cocci. These are also non-encapsulated, non-endospore-forming, non-halophilic, mesophilic, non-motile and Voges-Proskauer (production of indole and acetoin)-negative, and do not possess mycolic or teichoic acids. Kocuria species can be differentiated from other members of the Actinomycetales based on the presence of galactosamine and glucosamine as main cell wall amino sugars, the peptidoglycan type L-Lys-Ala 3/4 , the fatty acid anteisio-C 15 : 0 , the polar lipids diphosphatidylglycerol and phosphatidylglycerol, MK-7(H 2 ) and MK-8(H 2 ) as major menaquinones and a DNA G+C content of 60.0-75.3 mol%, depending on the species. Organisms in the genus are environmental bacteria, as well as human skin and oropharynx mucosa commensals; nevertheless, they can be responsible for infectious processes which mostly complicate severe underlying diseases. Owing to misidentification by phenotypic typing over the years, clinical syndromes caused by these agents are believed to be rare; however, the prevalence of such infectious pathologies is presumably higher and will surely increase in the coming years, as soon as genome-based identification i...

BackgroundStenotrophomonas maltophilia is emerging as one of the most frequently found bacteria in cystic fibrosis (CF) patients. In the present study, phenotypic and genotypic traits of a set of 98 isolates of S. maltophilia obtained from clinical (CF and non-CF patients) and environmental sources were comparatively evaluated.ResultsS. maltophilia exhibited a high level of genomic diversity in both CF and non-CF group, thus possibly allowing this bacterium to expand its pathogenic potentials. Strains sharing the same pulsotype infected different patients, thus likely indicating the occurrence of clonal spread or acquisition by a common source. CF isolates differed greatly in some phenotypic traits among each other and also when compared with non-CF isolates, demonstrating increased mean generation time and susceptibility to oxidative stress, but reduced ability in forming biofilm. Furthermore, in CF isolates flagella- and type IV pili-based motilities were critical for biofilm development, although not required for its initiation. Sequential isogenic strains isolated from the same CF patient displayed heterogeneity in biofilm and other phenotypic traits during the course of chronic infection. CF and non-CF isolates showed comparable virulence in a mouse model of lung infection.ConclusionsOverall, the phenotypic differences observed between CF and non-CF isolates may imply different selective conditions and persistence (adaptation) mechanisms in a hostile and heterogeneous environment such as CF lung. Molecular elucidation of these mechanisms will be essential to better understand the selective adaptation in CF airways in order to design improved strategies useful to counteract and eradicate S. maltophilia infection.

BackgroundStenotrophomonas maltophilia has recently gained considerable attention as an important emerging pathogen in cystic fibrosis (CF) patients. However, the role of this microorganism in the pathophysiology of CF lung disease remains largely unexplored. In the present study for the first time we assessed the ability of S. maltophilia CF isolates to adhere to and form biofilm in experimental infection experiments using the CF-derived bronchial epithelial IB3-1cell line. The role of flagella on the adhesiveness of S. maltophilia to IB3-1 cell monolayers was also assessed by using fliI mutant derivative strains.ResultsAll S. maltophilia CF isolates tested in the present study were able, although at different levels, to adhere to and form biofilm on IB3-1 cell monolayers. Scanning electron and confocal microscopy revealed S. maltophilia structures typical of biofilm formation on bronchial IB3-1 cells. The loss of flagella significantly (P < 0.001) decreased bacterial adhesiveness, if compared to that of their parental flagellated strains. S. maltophilia CF isolates were also able to invade IB3-1 cells, albeit at a very low level (internalization rate ranged from 0.01 to 4.94%). Pre-exposure of IB3-1 cells to P. aeruginosa PAO1 significantly increased S. maltophilia adhesiveness. Further, the presence of S. maltophilia negatively influenced P. aeruginosa PAO1 adhesiveness.ConclusionsThe main contribution of the present study is the finding that S. maltophilia is able to form biofilm on and invade CF-derived IB3-1 bronchial epithelial cells, thus posing a rationale for the persistence and the systemic spread of this opportunistic pathogen in CF patients. Experiments using in vivo models which more closely mimic CF pulmonary tissues will certainly be needed to validate the relevance of our results.

We tested 40 clinical Stenotrophomonas maltophilia strains to investigate the possible correlation between adherence to and formation of biofilm on polystyrene, and cell surface properties such as hydrophobicity and motility. Most of the strains were able to adhere and form biofilm, although striking differences were observed. Eleven (27.5%) of the strains were hydrophobic, with hydrophobicity greatly increasing as S. maltophilia attached to the substratum. A positive correlation was observed between hydrophobicity and levels of both adhesion and biofilm formation. Most of the isolates showed swimming and twitching motility. A highly significant negative correlation was observed between swimming motility and level of hydrophobicity. Hydrophobicity is thus a significant determinant of adhesion and biofilm formation on polystyrene surfaces in S. maltophilia.

Candida guilliermondii (teleomorph Meyerozyma guilliermondii) is an ascomycetous species belonging to the Saccharomycotina CTG clade which has been studied over the last 40 years due to its biotechnological interest, biological control potential and clinical importance. Such a wide range of applications in various areas of fundamental and applied scientific research has progressively made C. guilliermondii an attractive model for exploring the potential of yeast metabolic engineering as well as for elucidating new molecular events supporting pathogenicity and antifungal resistance. All these research fields now take advantage of the establishment of a useful molecular toolbox specifically dedicated to C. guilliermondii genetics including the construction of recipient strains, the development of selectable markers and reporter genes and optimization of transformation protocols. This area of study is further supported by the availability of the complete genome sequence of the reference strain ATCC 6260 and the creation of numerous databases dedicated to gene ontology annotation (metabolic pathways, virulence, and morphogenesis). These genetic tools and genomic resources represent essential prerequisites for further successful development of C. guilliermondii research in medical mycology and in biological control by facilitating the identification of the multiple factors that contribute to its pathogenic potential. These genetic and genomic advances should also expedite future practical uses of C. guilliermondii strains of biotechnological interest by opening a window into a better understanding of the biosynthetic pathways of valuable metabolites.