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1.  New insights in Staphylococcus pseudintermedius pathogenicity: antibiotic-resistant biofilm formation by a human wound-associated strain 
BMC Microbiology  2015;15:109.
Staphylococcus pseudintermedius is an opportunistic pathogen recognized as the leading cause of skin, ear, and post-operative bacterial infections in dogs and cats. Zoonotic infections have also recently been reported causing endocarditis, infection of surgical wounds, rhinosinusitis, and catheter-related bacteremia. The aim of the present study is to evaluate, for the first time, the pathogenic potential of S. pseudintermedius isolated from a human infection. To this end, strain DSM 25713, which was recently isolated from a wound of a leukemic patient who underwent a bone marrow transplantation, was investigated for biofilm formation and antibiotic-resistance under conditions relevant for wound infection.
The effect of pH (5.5, 7.1, and 8.7) and the presence of serum (diluted at 1:2, 1:10, and 1:100) on biofilm formation was assessed through a crystal violet assay. The presence of serum significantly reduced the ability to form biofilm, regardless of the pH value tested. In vitro activity of eight antibiotics against biofilm formation and mature 48 h-old biofilms was comparatively assessed by crystal violet assay and viable cell count, respectively. Antibiotics at sub-inhibitory concentrations reduced biofilm formation in a dose-dependent manner, although cefoxitin was the most active, causing a significant reduction already at 1/8xMIC. Rifampicin showed the highest activity against preformed biofilms (MBEC90: 2xMIC). None of the antibiotics completely eradicated the preformed biofilms, regardless of tested concentrations. Confocal and electron microscopy analyses of mature biofilm revealed a complex “mushroom-like” architecture consisting of microcolonies embedded in a fibrillar extracellular matrix.
For the first time, our results show that human wound-associated S. pseudintermedius is able to form inherently antibiotic-resistant biofilms, suggestive of its pathogenic potential, and consistent with recent reports of zoonotic infections.
PMCID: PMC4440327  PMID: 25994406
Staphylococcus pseudintermedius; zoonotic infection; Biofilm formation; Wound infection; Antibiotic-resistance
2.  Beta-Hemolytic, Multi-Lancefield Antigen-Agglutinating Enterococcus durans from a Pregnant Woman, Mimicking Streptococcus agalactiae 
Journal of Clinical Microbiology  2014;52(6):2181-2182.
A beta-hemolytic Lancefield antigen A-, B-, C-, D-, F-, and G-positive Enterococcus durans strain was cultivated from the rectovaginal swab of a pregnant woman who underwent antenatal screening for Streptococcus agalactiae. The isolate raised concern as to what extent similar strains are misrecognized and lead to false diagnosis of group B streptococci.
PMCID: PMC4042790  PMID: 24671782
3.  Small Colony Variant of Methicillin-Resistant Staphylococcus pseudintermedius ST71 Presenting as a Sticky Phenotype 
Journal of Clinical Microbiology  2014;52(4):1225-1227.
We first observed the phenomenon of small colony variants (SCVs) in a Staphylococcus pseudintermedius sequence type 71 (ST71) strain, isolated from a non-pet owner. Although we found that small-sized colonies share main features with Staphylococcus aureus SCVs, they nevertheless show a novel, particular, and sticky phenotype, whose expression was extremely stable, even after subcultivation.
PMCID: PMC3993511  PMID: 24452163
4.  Liofilchem® Chromatic VRE and vancomycin MIC Test Strip detected glycopeptide resistance in a vanB neonatal Enterococcus faecium isolate showing alternate vancomycin susceptibility and resistance with bioMérieux Vitek2 
A 1-month old neonate urine sample yielded vanB Enterococcus faecium; nevertheless, the isolate alternatively showed susceptibility and resistance to vancomycin with bioMérieux Vitek2 (cards AST592, AST632, AST586), while glycopeptide resistance was detected by Liofilchem® vancomycin MIC Test Strip and disc along with the Chromatic VRE chromogenic medium. This communication emphasizes that, as vanB gene may be heterogeneously expressed within a given Enterococcus population, glycopeptide resistance may be missed when using automated systems for antibiotic susceptibility testing. We suggest therefore that vancomycin in vitro activity be studied on all clinical isolates through agar methods, including use of chromogenic media.
PMCID: PMC4203251  PMID: 25337280
Enterococcus faecium; vanB; vancomycin; glycopeptide; neonate; newborn
5.  Successful off-label use of the Cepheid Xpert GBS in a late-onset neonatal meningitis by Streptococcus agalactiae  
We report the case of a late-onset neonatal meningitis by Streptococcus agalactiae (group B Streptococcus - GBS) that was diagnosed with a latex agglutination assay (on cerebrospinal fluid, CSF), as well as by using, for the first time, Xpert GBS (Cepheid, US) on CSF. Due to empirical antibiotics given before sampling, both CSF and blood culture were negative, so the abovementioned diagnostics was crucial. Moreover, the Xpert GBS assay, performed according to an off-label, modified protocol (the system is designed for GBS-carriage intrapartum screening, based on a completely automated real time-Polymerase Chain Reaction) quickly detected the organism’s genome target. Although further investigation on this test’s performace on CSF is required, then, we trust it may be a promising, quick and precise diagnostic method for infections in newborns.
PMCID: PMC4152086  PMID: 25197396
Streptococcus agalactiae; GBS; Lancefield group B; pregnancy; gestation; meningitis; Cepheid; Xpert GBS
6.  Surgical wound infection by mannitol-nonfermenting Staphylococcus aureus after lumbar microdiscectomy 
Purulent infection of a surgical wound developed after discectomy, and a mannitol-nonfermenting Staphylococcus aureus isolate was cultivated as the etiologic agent. Nonfermenting S. aureus strains are exceedingly rare and may be erroneously mistaken and dismissed as contaminants. This report then emphasizes that pure and massive cultures must be carefully evaluated, even if preliminary examination does not suggest a pathogenic organism. Also, although mannitol-negative, the studied strain was correctly detected as S. aureus by both the-FISH test (AdvanDx, USA) and the Liofilchem ‘Chromatic Staph aureus’, highlighting that additional diagnostic methods may support recognition of uncommon, nonfermenting S. aureus strains in the daily practice.
PMCID: PMC4069970  PMID: 24966984
Mannitol; Staphylococcus aureus; surgical wound
7.  Staphylococcus pseudintermedius for CAMP-test 
CAMP test reliably detects Listeria monocytogenes (Lm) and Streptococcus agalactiae (group B streptococcus, GBS); it is traditionally performed streaking the tested isolate perpendicularly to Staphylococcus aureus (Sa), provided that reference Sa strains (that produce β-hemolysin) are used. In a zone of β-hemolysin activity, in fact, GBS and Lm form typical arrow-shaped hemolytic areas. While Sa production of the toxin is strain-dependent, however, that of Staphylococcus pseudintermedius (Sp), a pet-owner colonizer and an emerging human pathogen, is constitutive, then observed in all clinical isolates. Therefore, Sp may indeed represent a valid alternative to perform the assay.
PMCID: PMC4014255  PMID: 24817971
Staphylococcus pseudintermedius; β-hemolysin; CAMP-test; Listeria monocytogenes; GBS
8.  Liofilchem® O.A. Listeria agar and direct CAMP test provided sooner Listeria monocytogenes identification from neonatal bacteremia 
Listeria monocytogenes infection in pregnant women and newborns is a cause for serious concern, and invasive disease outcome strongly depends on prompt antibiotic therapy. To provide sooner identification from neonatal bacteremia we performed a CAMP test directly on positive blood aliquots and inoculated the Liofilchem® O.A. Listeria chromogenic agar as well, thus providing a 24-h turn-around time for response.
PMCID: PMC3971323  PMID: 24695762
Listeria monocytogenes; neonate; newborn; pregnancy; CAMP test; Liofilchem® O.A. Listeria agar
9.  Methicillin-Resistant Staphylococcus pseudintermedius Infection in a Bone Marrow Transplant Recipient 
Journal of Clinical Microbiology  2013;51(5):1636-1638.
Staphylococcus pseudintermedius is a veterinary pathogen that has seldom been described as an agent of human disease. Features of this probably underreported coagulase-positive Staphylococcus species are depicted here through the description of a graft-versus-host disease-related wound infection caused by a multidrug-resistant strain.
PMCID: PMC3647946  PMID: 23486715
10.  Streptococcus agalactiae vaginitis: nonhemolytic variant on the Liofilchem® Chromatic StreptoB 
Streptococcus agalactiae (group B Streptococcus, GBS) vaginal pathogenicity is not uniformly acknowledged throughout the literature; accordingly, in women, genital itching and burning, along with leukorrhea are commonly and almost exclusively referred to bacterial vaginosis, candidiasis and trichomoniasis. Conversely, GBS virulence for vagina was recognized in the past, as the organism has been observed to potentially cause local inflammation and discharge, as well as lactobacilli rarefaction. We depict here a case where a nonhemolytic (γ-hemolytic) GBS strain was found to be the etiologic agent of vaginal infection. Such uncommon S. agalactiae phenotypes are hard to be recognized and may be therefore responsible for misdiagnosing and underestimation of GBS vaginitis prevalence; here, we had the support of the Liofilchem® Chromatic StreptoB medium, that successfully detected such an atypical variant.
PMCID: PMC3726989  PMID: 23923091
Streptococcus agalactiae; group B antigen; GBS; vaginitis; leukorrhea
11.  Preliminary Evaluation of the Safety and Efficacy of Standard Intravenous Immunoglobulins in Pregnant Women with Primary Cytomegalovirus Infection 
Clinical and Vaccine Immunology : CVI  2012;19(12):1991-1993.
Hyperimmune globulins were reported to prevent and treat fetal cytomegalovirus (CMV) infection during pregnancy. Here, we report that infusions of standard human intravenous immunoglobulin significantly increase CMV IgG titers and avidity indexes in pregnant women, paving the way to their use for passive transfer of maternal CMV humoral immunity to fetuses. Preliminary data on perinatal outcomes of the first 67 newborns are encouraging.
PMCID: PMC3535874  PMID: 23100477
13.  Successful salvage therapy with Daptomycin for osteomyelitis caused by methicillin-resistant Staphylococcus aureus in a renal transplant recipient with Fabry-Anderson disease 
Daptomycin is licensed in adults for the management of Staphylococcus aureus methicillin-resistant infections, including bone and skin complicated infections. We describe for the first time its use in a renal transplant recipient for Fabry-Anderson Disease with right heel osteomyelitis. The patient was unresponsive to first-line Teicoplanin and second-line Tigecycline, whereas he was successfully treated with third-line Daptomycin monotherapy at 4 mg/Kg/qd for 4 weeks. Local debridement was performed in advance of each line of treatment.
PMCID: PMC3324387  PMID: 22404900
Methicillin-resistant Staphylococcus aureus; Osteomyelitis; Daptomycin; Salvage therapy; Antibiotic therapy
14.  Phenotypic and genotypic characterization of Stenotrophomonas maltophilia isolates from patients with cystic fibrosis: Genome diversity, biofilm formation, and virulence 
BMC Microbiology  2011;11:159.
Stenotrophomonas 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.
S. 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.
Overall, 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.
PMCID: PMC3146419  PMID: 21729271
15.  Brief Tale of a Bacteraemia by Rhodococcus equi, With Concomitant Lung Mass: What Came First, the Chicken or The Egg? 
Rhodococcus equi is an uncommon Gram positive, variably acid-fast pathogen, that appears as hard to treat mostly owing to the establishment of intracellular niches. Lack of interpretive criteria for susceptibility testing may lead to under-reporting or overestimation of resistances, whereas knowledge about this pathogen’s clinical impact may be affected by erroneous phenotype-based characterization at a genus and species level.
We present the case of a bacteraemia with a concomitant lung mass in a lymphoma patient, that further highlights the emergence of rhodococcal diseases as a matter for concern in the fields of infectious diseases and haematology.
PMCID: PMC3103262  PMID: 21625310
16.  Adhesion to and biofilm formation on IB3-1 bronchial cells by Stenotrophomonas maltophilia isolates from cystic fibrosis patients 
BMC Microbiology  2010;10:102.
Stenotrophomonas 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.
All 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.
The 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.
PMCID: PMC2858031  PMID: 20374629
17.  Femoral Prosthesis Infection by Rhodotorula mucilaginosa▿  
Journal of Clinical Microbiology  2008;46(10):3544-3545.
This case report is a case history of a femoral prosthesis infection caused by Rhodotorula mucilaginosa in a human immunodeficiency virus patient. Though the pathogenicity of this organism for bone tissue has been previously reported, this is the first reported case of an orthopedic prosthesis infection by this species of the genus Rhodotorula.
PMCID: PMC2566075  PMID: 18753353
18.  Multidrug-Resistant Escherichia fergusonii: a Case of Acute Cystitis▿  
Journal of Clinical Microbiology  2008;46(4):1551-1552.
We report a case in which Escherichia fergusonii, an emerging pathogen in various types of infections, was associated with cystitis in a 52-year-old woman. The offending strain was found to be multidrug resistant. Despite in vitro activity, beta-lactam treatment failed because of a lack of patient compliance with therapy. The work confirms the pathogenic potential of E. fergusonii.
PMCID: PMC2292955  PMID: 18256229
19.  β-Lactam Failure in Treatment of Two Group G Streptococcus dysgalactiae subsp. equisimilis Pharyngitis Patients▿  
Journal of Clinical Microbiology  2007;46(2):814-816.
We present two cases of exudative pharyngitis due to Streptococcus dysgalactiae subsp. equisimilis, Lancefield group G. While the participation of this organism as an agent of pharyngitis is well documented, we focus on failure of beta-lactam therapy, a phenomenon that is well described for pharyngitis due to Streptococcus pyogenes. Therefore, these case reports add to our knowledge of pharyngitis caused by non-S. pyogenes streptococci.
PMCID: PMC2238097  PMID: 18057124

Results 1-19 (19)