Water disinfection by hydrodynamic cavitation in a rotor-stator device.

The efficiency of a rotor-stator device for water disinfection based on hydrodynamic cavitation is investigated. Water is infected with E. coli and E. faecalis with initial concentrations in the range 5 × 102-1.2 × 106 CFU/ml. Various geometries of the cavitation channel between rotor and stator are tested, achieving bacterial annihilation in less than 10 min of treatment times. Microorganism permanent elimination is verified via micro-seeding to discard viable non-culturable bacteria; micro-seeding was done for those samples displaying no CFU growth via normalized cultures on a Petri dish. TEM photographs are analyzed and the extent of bacterial damages is tentatively correlated with the various cavitation mechanisms. Rotor-stator cavitation assemblies used in the current research are between one and two orders of magnitude more energy efficient than those tested by other investigators. Acoustic pressure spectra are measured to assess the implosion intensity. Parametric analyses are conducted changing the rotor diameter (110-155 mm), the cavitation channel contraction ratio, Amax/Amin(4.56-5.0), and the number of contractions (Nr:58-80 rotor vanes; Ns:8-16 stator vanes).

[Structural dynamics of Legionella pneumophila and Legionella bozemanii colony/biofilm]. / Dinámica estructural de colonias/biofilm de Legionella pneumophila y Legionella bozemanii.

OBJECTIVES: The genus Legionella includes very pleomorphic species responsible for disease outbreaks in humans. The appearance of such has great importance to develop artificial biofilms in aquatic ecosystems. The aim of this work was to study the dynamics of growth and evolution of the internal structure of colonies of representative species of the genus as static biofilm model. METHODS: Isolated colonies of Legionella pneumophila and Legionella bozemanii grown in specific media for three and fifteen days were processed for histological methods and embedded in paraffin and epoxy resin for analysis by light microscopy, electron microscopy and image analysis. RESULTS. In colonies of both species were observed and defined specific architectural patterns, based on stratification and evolve over time. The strata differ in the amount of extracellular matrix, the morphology and population density and the proportion of dead cells. The internal structure of three days colonies showed large differences between L. pneumophila (two layers) and L. bozemanii (four layers). However, in the fifteen days colonies of both species evolved towards a common unique pattern formed by three layers. In both species the growth was also found within the culture medium, although this phenomenon was more intense in L. bozemanii with unique, central and larger invasions. CONCLUSIONS: Our results demonstrate that Legionella colonies on solid culture media are a good model of static biofilm with a complex structural dynamics characterized by the presence of morphological and functional subpopulations. We bring here an histological approach model, allowing, in further research, detailed studies in evolutionary adaptations in multicellular communities to adverse media and to antimicrobials in Legionella species of clinical interest.

Dinámica estructural de colonias/biofilm de Legionella pneumophila y Legionella bozemanii / Structural dynamics of Legionella pneumophila and Legionella bozemanii colony/biofilm

Objetivos. El género Legionella engloba especies muy pleomórficas responsables de brotes infecciosos en humanos. En la aparición de los mismos tiene gran importancia el desarrollo de biofilms en ecosistemas acuáticos artificiales. El objetivo de este trabajo fue estudiar la dinámica de crecimiento y la evolución de la estructura interna de colonias de especies representativas del género como modelo de biofilm estático. Material y métodos. Colonias aisladas de Legionella pneumophila y Legionella bozemanii crecidas en medios específicos durante tres y quince días fueron procesadas por métodos histológicos de inclusión en parafina y resina epoxi para su análisis mediante microscopía óptica, microscopía electrónica y análisis de imagen. Resultados. En las colonias de ambas especies se observaron patrones arquitecturales definidos y específicos, basados en la estratificación y que evolucionan en el tiempo. Los estratos se diferencian por la cantidad de matriz extracelular, la morfología y densidad poblacional y la proporción de células muertas. La estructura interna de las colonias de tres días presentaba grandes diferencias entre L. pneumophila (dos estratos) y L. bozemanii (cuatro estratos). Sin embargo, en las colonias de quince días ambas especies evolucionaron hacia un patrón único común formado por tres estratos. En ambas especies se comprobó también el crecimiento en el interior del medio de cultivo, aunque este fenómeno fue mucho más intenso en L. bozemanii, con invasiones únicas, centrales y de gran tamaño. Conclusiones. Nuestros resultados demuestran que las colonias de Legionella sobre medio de cultivo sólido son un buen modelo de biofilm estático, con una dinámica estructural compleja caracterizada por la presencia de subpoblaciones morfológicas y funcionales. La aproximación histológica empleada en este modelo permitirá estudiar adaptaciones evolutivas de comunidades multicelulares a medios hostiles, así como la respuesta a los antimicrobianos de las especies de Legionella de interés clínico (AU)

Objectives. The genus Legionella includes very pleomorphic species responsible for disease outbreaks in humans. The appearance of such has great importance to develop artificial biofilms in aquatic ecosystems. The aim of this work was to study the dynamics of growth and evolution of the internal structure of colonies of representative species of the genus as static biofilm model. Methods. Isolated colonies of Legionella pneumophila and Legionella bozemanii grown in specific media for three and fifteen days were processed for histological methods and embedded in paraffin and epoxy resin for analysis by light microscopy, electron microscopy and image analysis. Results. In colonies of both species were observed and defined specific architectural patterns, based on stratification and evolve over time. The strata differ in the amount of extracellular matrix, the morphology and population density and the proportion of dead cells. The internal structure of three days colonies showed large differences between L. pneumophila (two layers) and L. bozemanii (four layers). However, in the fifteen days colonies of both species evolved towards a common unique pattern formed by three layers. In both species the growth was also found within the culture medium, although this phenomenon was more intense in L. bozemanii with unique, central and larger invasions. Conclusions. Our results demonstrate that Legionella colonies on solid culture media are a good model of static biofilm with a complex structural dynamics characterized by the presence of morphological and functional subpopulations. We bring here an histological approach model, allowing, in further research, detailed studies in evolutionary adaptations in multicellular communities to adverse media and to antimicrobials in Legionella species of clinical interest (AU)

Evaluation of bacterial resistance to essential oils and antibiotics after exposure to oregano and cinnamon essential oils.

Essential oils (EOs) are excellent antimicrobial agents sometimes used in active food packaging. This work studies the susceptibility of 48 clinical isolates and 12 reference strains of Gram-negative bacilli to oregano essential oil, cinnamon essential oil, and combinations of both. Furthermore, the tendency of the clinical isolates to develop resistance to these EOs and to different antibiotics after sequential oregano or cinnamon exposure was studied. For this purpose, antibiotic susceptibility (through disk diffusion assays and minimum inhibitory concentration [MIC] determination) and oregano and cinnamon susceptibility (through MIC and minimum bactericidal concentration [MBC] determination) were compared after 50 passages in the presence or absence of subinhibitory concentrations of oregano and cinnamon essential oils. The results showed that all strains were susceptible to both EOs and their combination independently of the antibiotic resistance profile. In addition, neither synergistic nor antagonistic effects were observed between oregano and cinnamon essential oils at the concentrations tested. After the sequential exposure to both EOs, only Serratia marcescens, Morganella morganii, and Proteus mirabilis treated with oregano changed their antibiotic resistance profile and/or increased their resistance to this EO. However, the changes in antibiotic and oregano resistance were not related.

Characterization of the enzyme aac(3)-id in a clinical isolate of Salmonella enterica serovar Haifa causing traveler's diarrhea

Introducciónel objetivo de este estudio fue identificar el mecanismo de sensibilidad disminuida a gentamicina en un aislamiento clínico de Salmonella, lo que nos condujo a la detección de un gen que codifica una acetiltransferasa modificante de aminoglucósidos localizada en un integron tipo 1.Métodosla cepa multiresistente de Salmonella fue aislada de las heces de un viajero a Egipto. La susceptibilidad a 12 agentes antimicrobianos se determinó mediante Kirby-Bauer. La presencia de integron clase 1 se realizó mediante PCR. El producto de PCR amplificado del integrón fue recuperado y secuenciado para conocer los genes que contenía dicho integrón. Además se determinó la susceptibilidad a gentamicina C1a, gentamicina C1, sisomicina, neomicina, dibekacina, kanamicina, tobramicina, amikacina, netilmicina, apramicina, dactimicina, espectinomicina, estreptomicina, lividomicina y butirosina. El kit de expresión Champion™ pET101 Directional TOPO® fue utilizado para clonar y expresar el gen aac(3)-I.Resultadosel aislamiento fue identificado como Salmonella enterica serovariedad Haifa, el cual presentaba resistencia al ácido nalidixico, tetraciclina y sensibilidad disminuida a gentamicina. Se observó la presencia de un integron tipo 1 con un tamaño de 1,500bp en el que se encontraron dos genes (aac(3)-Id y aadA7). El análisis de la sensibilidad a diferentes aminoglucósidos de la cepa de E. coli TOP10F’ transformada con el vector que contenia el gen aac(3)-Id demostró resistencia a gentamicina C1a, gentamicina C1, y dactimicina, en concordancia con la presencia del enzima pero era susceptible a sisomicina. La secuencia de aminoácidos presentaba un 100% de identidad con el enzima AAC(3)-Id.Conclusiónla presencia del enzima AAC(3)-Id ha sido descrita por primera vez en S. Haifa(AU)

IntroductionThe objective of this investigation was to identify the mechanism of decreased susceptibility to gentamicin in a Salmonella clinical isolate, leading to the detection of a aminoglycoside acetyltransferase gene found in a class 1 integron.MethodsA multidrug-resistant Salmonella strain was recovered from feces of a traveler to Egypt. The antimicrobial susceptibility test to 12 antimicrobial agents was performed with the Kirby-Bauer method. The presence of class 1 integron was determined by PCR. The amplified product was recovered and sequenced in order to establish the genes carried. In addition, susceptibility to gentamicin C1a, gentamicin C1, sisomicin, neomycin, dibekacin, kanamycin, tobramycin, amikacin, netilmicin, apramycin, dactimicin, spectinomycin, streptomycin, lividomycin and butirosin, was established. The Champion™ pET101 Directional TOPO® Expression Kit was used to clone and express the aac(3)-I gene.ResultsThe isolate was identified as Salmonella enterica serovar Haifa, showing resistance to nalidixic acid, tetracycline and decreased susceptibility to gentamicin. One integron with a size circa 1,500bp, encoding an aac(3)-Id plus aadA7 genes was observed. The analysis of the susceptibility to different aminoglycosides in the E. coli TOP10F’ transformed with the vector carrying aac(3)-Id gene showed resistance to gentamicin C1a, gentamicin C1, and dactimicin, in accordance with the presence of this enzyme but, was susceptible to sisomicin. The homology of the amino acid and nucleotide sequences with the AAC(3)-Id enzyme was of 100%.ConclusionThe presence of the AAC(3)-Id enzyme was described for the first time in a S. Haifa(UA)

Characterization of the enzyme aac(3)-Id in a clinical isolate of Salmonella enterica serovar Haifa causing traveler's diarrhea.

INTRODUCTION: The objective of this investigation was to identify the mechanism of decreased susceptibility to gentamicin in a Salmonella clinical isolate, leading to the detection of a aminoglycoside acetyltransferase gene found in a class 1 integron. METHODS: A multidrug-resistant Salmonella strain was recovered from feces of a traveler to Egypt. The antimicrobial susceptibility test to 12 antimicrobial agents was performed with the Kirby-Bauer method. The presence of class 1 integron was determined by PCR. The amplified product was recovered and sequenced in order to establish the genes carried. In addition, susceptibility to gentamicin C1a, gentamicin C1, sisomicin, neomycin, dibekacin, kanamycin, tobramycin, amikacin, netilmicin, apramycin, dactimicin, spectinomycin, streptomycin, lividomycin and butirosin, was established. The Champion pET101 Directional TOPO Expression Kit was used to clone and express the aac(3)-I gene. RESULTS: The isolate was identified as Salmonella enterica serovar Haifa, showing resistance to nalidixic acid, tetracycline and decreased susceptibility to gentamicin. One integron with a size circa 1,500 bp, encoding an aac(3)-Id plus aadA7 genes was observed. The analysis of the susceptibility to different aminoglycosides in the E. coli TOP10F' transformed with the vector carrying aac(3)-Id gene showed resistance to gentamicin C1a, gentamicin C1, and dactimicin, in accordance with the presence of this enzyme but, was susceptible to sisomicin. The homology of the amino acid and nucleotide sequences with the AAC(3)-Id enzyme was of 100%. CONCLUSION: The presence of the AAC(3)-Id enzyme was described for the first time in a S. Haifa.

In vitro activity of cefditoren and other antimicrobial agents against 288 Streptococcus pneumoniae and 220 Haemophilus influenzae clinical strains isolated in Zaragoza, Spain.

In vitro cefditoren antimicrobial activity was tested against 288 Streptococcus pneumoniae and 220 Haemophilus influenzae clinical strains isolated in our hospital from January 2005 to May 2006 by agar dilution and broth microdilution method, respectively. MICs were also determined for 13 and 10 comparison drugs, respectively. The pneumococci tested comprised 113 (39.2%) penicillin susceptible, 91 (31.6%) penicillin intermediate, and 84 (29.2%) penicillin resistant. Cefditoren was the most active drug on the basis of the MICs (MIC(90)=0.5 microg/mL), followed by ceftriaxone and levofloxacin (MIC(90)=1 microg/mL). Cefditoren MICs ranged from 0.25 to 1 microg/mL for ceftriaxone-resistant isolates, with a modal MIC of 0.5 microg/mL and an MIC(90) of 1.0 microg/mL. No S. pneumoniae isolates evaluated in this study showed MICs to cefditoren higher than 1 microg/mL (MIC range, 4 microg/mL). Against H. influenzae (Hi beta+), the rank order of intrinsic activity (MIC(90), microg/mL) was cefditoren (0.03) < cefixime (0.06)8.0).

[Susceptibility to betalactam antibiotics, glycopeptides and aminoglycosides in commensal strains of erythromycin-resistant alpha-hemolytic streptococci and Gemella spp]. / Sensibilidad a antibióticos betalactámicos, glucopéptidos y aminoglucósidos en cepas comensales de estreptococos alfahemolíticos y Gemella spp. resistentes a eritromicina.

OBJECTIVE: Susceptibility to seven betalactam antibiotics, glycopeptides and aminoglycosides was investigated in 190 erythromycin-resistant alpha-hemolytic streptococci and 30 Gemella spp, mainly from normal flora. MATERIAL AND METHODS: Antimicrobial susceptibility testing was performed by a standard agar diffusion test and a standard agar dilution method according to NCCLS/CLSI criteria. RESULTS: 62.6% of alfahemolytic streptococci and 53.3% of Gemella spp. were not susceptible to penicillin (MIC50: 0,5 microg/mL). Cefuroxime was the least active cephalosporin (MIC50: 1 microg/mL and 0.5 microg/mL, in streptococci and Gemella spp., respectively), whereas cefotaxime, ceftriaxone (MIC50: 0.25 microg/mL) and cefepime (MIC50: 0.5 microg/mL) were more active than penicillin. All isolates were susceptible to vancomycin, teicoplanin and gentamicin. Four alfahemolytic streptococcal strains showed high-level resistance to streptomycin, and three strains to kanamycin. There were no significant differences in resistance rates to the antibiotics studied between strains with different macrolide resistance phenotypes. Resistance to penicillin and other betalactam antibiotics (73.8%) was prevalent in M phenotype strains and resistance to penicillin and other classes of antibiotics predominated in constitutive (cMLS(B)) strains (71.4%). CONCLUSIONS: Resistance to penicillin in erythromycin-resistant strains was notably high in this study. This fact has important clinical implications because of the endogenous character of alpha-hemolytic streptococcal and Gemella spp. infections. The lower cefuroxime activity suggests that use of this agent against other pathogens would be effective in preserving the oropharyngeal microflora analyzed.

Sensibilidad a antibióticos betalactámicos, glucopéptidos y aminoglucósidos en cepas comensales de estreptococos alfahemolíticos y Gemella spp. resistentes a eritromicina / Susceptibility to betalactam antibiotics, glycopeptides and aminoglycosides in commensal strains of erythromycin resistant alfahemolytic streptococci and Gemella spp

Objetivo. Se investigó la sensibilidad de 190 estreptococos alfahemolíticos y 30 Gemella spp., en su mayoría comensales y resistentes a eritromicina, frente a 7 antibióticos betalactámicos, glucopépticos y aminoglucósidos. Material y métodos. Para determinar la sensibilidad a los antimicrobianos se utilizaron las técnicas de dilución y difusión en agar, según las normas del National Committee for Clinical Laboratory Standards/Clinical and Laboratory Standards Institute (NCCLS/CLSI). Resultados. El 62,6% de los estreptococos alfahemolíticos y el 53,3% de Gemella spp. no fueron sensibles a penicilina (concentración inhibitoria mínima al 50% [CIM50]: 0,5 mg/ml). Cefuroxima fue la cefalosporina que presentó menor actividad (CIM50: 1 mg/ml y 0,5 mg/ml, en estreptococos y Gemella spp.), mientras que cefotaxima, ceftriaxona (CIM50: 0,25 mg/ml) y cefepima (CIM50: 0,5 mg/ml) presentaron una actividad superior a penicilina. El 100% de los aislamientos fueron sensibles a vancomicina, teicoplanina y gentamicina. Cuatro cepas de estreptococos alfahemolíticos presentaron resistencia de alto nivel a estreptomicina y tres a kanamicina. No hubo diferencia significativa en el comportamiento de los antibióticos estudiados frente a las cepas con diferente fenotipo de resistencia a macrólidos. Tanto en los aislamientos con fenotipo M como en aquéllos con fenotipo constitutivo (MLSBc) predominó la resistencia a penicilina y a otros antibióticos betalactámicos frente al patrón de resistencia, que incluye sólo penicilina o bien penicilina, otros antibióticos betalactámicos y aminoglucósidos. Conclusiones. Las tasas de resistencia a penicilina en las cepas comensales resistentes a eritromicina son particularmente elevadas, hecho que tiene una importante implicación clínica por el carácter endógeno de las infecciones causadas por estas bacterias. La menor actividad de cefuroxima podría sugerir que su utilización frente a otros patógenos preservaría en mayor grado a la microbiota orofaríngea estudiada (AU)

Objective. Susceptibility to seven betalactam antibiotics, glycopeptides and aminoglycosides was investigated in 190 erythromycin-resistant alfahemolytic streptococci and 30 Gemella spp, mainly from normal flora. Material and methods. Antimicrobial susceptibility testing was performed by a standard agar diffusion test and a standard agar dilution method according to NCCLS/CLSI criteria. Results. 62.6% of alfahemolytic streptococci and 53.3% of Gemella spp. were not susceptible to penicillin (MIC50: 0,5 mg/mL). Cefuroxime was the least active cephalosporin (MIC50: 1 mg/mL and 0.5 mg/mL, in streptococci and Gemella spp., respectively), whereas cefotaxime, ceftriaxone (MIC50: 0.25 mg/mL) and cefepime (MIC50: 0.5 mg/mL) were more active than penicillin. All isolates were susceptible to vancomycin, teicoplanin and gentamicin. Four alfahemolytic streptococcal strains showed high-level resistance to streptomycin, and three strains to kanamycin. There were no significant differences in resistance rates to the antibiotics studied between strains with different macrolide resistance phenotypes. Resistance to penicillin and other betalactam antibiotics (73.8%) was prevalent in M phenotype strains and resistance to penicillin and other classes of antibiotics predominated in constitutive (cMLSB) strains (71.4%). Conclusions. Resistance to penicillin in erythromycin-resistant strains was notably high in this study. This fact has important clinical implications because of the endogenous character of alfahemolytic streptococcal and Gemella spp. infections. The lower cefuroxime activity suggests that use of this agent against other pathogens would be effective in preserving the oropharyngeal microflora analyzed (AU)

Molecular typing of Staphylococcus aureus clinical isolates by pulsed-field gel electrophoresis, staphylococcal cassette chromosome mec type determination and dissemination of antibiotic resistance genes.

Pulsed-field gel electrophoresis (PFGE) of SmaI macrorestriction fragments of genomic DNA as well as staphylococcal cassette chromosome mec (SCCmec) typing for mecA-carrying isolates were used to study the distribution of clonal types among 177 Staphylococcus aureus clinical isolates recovered in a Spanish hospital between 2000 and 2003. Five major clonal types (P1 to P5) were identified by PFGE, with one of them (P1) comprising the majority of strains (47.5%). According to SCCmec typing, SCCmec type IVA was the most prevalent type, showing increasing prevalence in the hospital setting with respect to other pandemic clones. One SCCmec pattern was detected in different PFGE types, which demonstrates that the latter is a major discriminative typing method. Three novel SCCmec elements or variants were found, each in a different PFGE type. Oxacillin (methicillin)-resistant and -susceptible S. aureus (MRSA and MSSA, respectively) strains were detected showing identical PFGE patterns, suggesting horizontal transfer of mecA to MSSA and/or mecA deletion from MRSA. Persistence of several S. aureus clones throughout the years within the same hospital environment was also observed.

Novel 10-bp deletion in the translational attenuator of a constitutively expressed erm(A) gene from Staphylococcus epidermidis.

Alterations in the erm(A) regulatory region of six clinical isolates of Staphylococcus epidermidis and one of Staphylococcus haemolyticus displaying a constitutive resistance phenotype were investigated. Anovel deletion of 10 bp with respect to the corresponding sequence of Tn554 was identified in the attenuator of a constitutively expressed erm(A) gene of one of the S. epidermidis isolates. Thus far, this is the smallest deletion conferring constitutive resistance in the translational attenuator of erm(A) in a naturally occurring S. epidermidis strain of human origin.

Novel 10-bp deletion in the translational attenuator of a constitutively expressed erm(A) gene from Staphylococcus epidermidis

Alterations in the erm(A) regulatory region of six clinical isolates of Staphylococcus epidermidis and one of Staphylococcus haemolyticus displaying a constitutive resistance phenotype were investigated. A novel deletion of 10 bp with respect to the corresponding sequence of Tn554 was identified in the attenuator of a constitutively expressed erm(A) gene of one of the S. epidermidis isolates. Thus far, this is the smallest deletion conferring constitutive resistance in the translational attenuator of erm(A) in a naturally occurring S. epidermidis strain of human origin (AU)

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Detection of the aminoglycosidestreptothricin resistance gene cluster ant(6)-sat4-aph(3 ')-III in commensal viridans group streptococci.

High-level aminoglycoside resistance was assessed in 190 commensal erythromycin-resistant alpha-hemolytic streptococcal strains. Of these, seven were also aminoglycoside-resistant: one Streptococcus mitis strain was resistant to high levels of kanamycin and carried the aph(3 ')-III gene, four S. mitis strains were resistant to high levels of streptomycin and lacked aminoglycoside-modifying enzymes, and two S. oralis strains that were resistant to high levels of kanamycin and streptomycin harbored both the aph(3 ')-III and the ant(6) genes. The two S. oralis strains also carried the ant(6)-sat4- aph(3 ' ')-III aminoglycoside-streptothricin resistance gene cluster, but it was not contained in a Tn5405-like structure. The presence of this resistance gene cluster in commensal streptococci suggests an exchange of resistance genes between these bacteria and enterococci or staphylococci.

Genetic characterization of Cryptosporidium species from humans in Spain.

Several species of Cryptosporidium have been associated with infection. Cryptosporidium parvum and Cryptosporidium hominis are the main agents of cryptosporidiosis in humans. Stool samples from 108 Cryptosporidium-infected patients were submitted to PCR-RFLP analysis for a 553-bp fragment of Cryptosporidium oocyst wall protein (COWP) gene and an 826-864 bp fragment of the small-subunit ribosomal RNA (SSU-rRNA) gene. Ninety-two patients were immunocompetent children and 16 were HIV-infected adults. C. hominis was detected in 69 patients (59 immunocompetent and 10 HIV-infected); C. parvum, in 34 patients (28 immunocompetent and 6 HIV-infected); and C. meleagridis and C. felis in one patient each (both immunocompetent children). Three samples yielded negative results. C. parvum was significantly more frequent in children from rural areas than in those of urban residence (p=0.010). As far as we know, this is the first surveillance study about the molecular characterization of Cryptosporidium in humans performed in Spain. The finding of zoonotic species infecting humans calls for further research on this subject.

Detection of the aminoglycosidestreptothricin resistance gene cluster ant(6)-sat4-aph(3')-III in commensal viridans group streptococci

High-level aminoglycoside resistance was assessed in 190 commensal erythromycin-resistant alpha-hemolytic streptococcal strains. Of these, seven were also aminoglycoside-resistant: one Streptococcus mitis strain was resistant to high levels of kanamycin and carried the aph(3’)-III gene, four S. mitis strains were resistant to high levels of streptomycin and lacked aminoglycoside-modifying enzymes, and two S. oralis strains that were resistant to high levels of kanamycin and streptomycin harbored both the aph(3’)-III and the ant(6) genes. The two S. oralis strains also carried the ant(6)-sat4- aph(3’’)-III aminoglycoside-streptothricin resistance gene cluster, but it was not contained in a Tn5405-like structure. The presence of this resistance gene cluster in commensal streptococci suggests an exchange of resistance genes between these bacteria and enterococci or staphylococci (AU)

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Molecular epidemiology of macrolide and tetracycline resistances in commensal Gemella sp. isolates.

The epidemiologic relatedness of 29 erythromycin-resistant Gemella sp. strains from normal flora, characterized previously, were evaluated by pulsed-field gel electrophoresis (PFGE). Three isolates carried the tet(O) gene and the tet(M) gene. The msr(A) gene was found in two Gemella morbillorum strains in combination with the erm(B) or mef(E) gene. The sequences of the mef(A/E), erm(B), and msr(A) genes showed a high similarity to the corresponding sequences of other gram-positive cocci. All the strains harboring the mef(A/E) gene and the msr(D) gene possessed open reading frame 3 (ORF3)/ORF6. The 16 G. morbillorum isolates represented 15 distinct DNA profiles. Four clusters were identified (>or=80% genetic relatedness). The 12 Gemella haemolysans strains belonged to different PFGE types. The clonal diversity found suggests that horizontal transfer may be the main route through which erythromycin resistance is acquired.

Novel streptomycin resistance gene from Mycobacterium fortuitum.

We have isolated the aph(3")-Ic gene, encoding an aminoglycoside 3"-O-phosphotransferase [APH(3")-Ic], from a genomic library of an environmental Mycobacterium fortuitum strain, selecting for streptomycin resistance. APH(3")-Ic phosphorylates and inactivates streptomycin. Similar genes have been described in Streptomyces griseus and plasmid RSF1010. It is also present in some M. fortuitum clinical isolates.

Enterococci spp. isolated from Cuba: species frequency of occurrence and antimicrobial susceptibility profile.

This study investigated the species distribution and antimicrobial resistance among 99 enterococci isolated from hospitalized patients in 12 hospitals in Cuba from October 2000 to September 2001. Species identification was performed by WIDER Automatic System (Francisco Soria Melguizo, Madrid, Spain), and the susceptibility testing was performed by disk diffusion, agar dilution, and E-test methods. Enterococcus faecalis was the most prevalent (85%) species, followed by E. faecium (10%), E. gallinarum (2%), E. casseliflavus (2%), and E. durans-hirae (1%). A higher percentage of resistance to ampicillin (50%), fosfomycin (40%), ciprofloxacin (30%), norfloxacin (20%), and tetracycline (90%) was detected in E. faecium isolates, whereas E. faecalis strains showed higher rates of resistance to erythromycin (52.4%), chloramphenicol (34.5%), rifampicin (62.5%), moxifloxacin (3%), and nitrofurantoin (2.4%). Resistance to glycopeptide was detected in E. faecalis (1.2%) and E. faecium (10%). Thirty-one E. faecalis (37%) and 3 E. faecium (30%) showed a high-level resistance to gentamicin. The results of this work will be very helpful to guide empirical antimicrobial therapy and the implementation of infection control measures in Cuban hospitals.

Molecular basis of resistance to macrolides and other antibiotics in commensal viridans group streptococci and Gemella spp. and transfer of resistance genes to Streptococcus pneumoniae.

We assessed the mechanisms of resistance to macrolide-lincosamide-streptogramin B (MLS(B)) antibiotics and related antibiotics in erythromycin-resistant viridans group streptococci (n = 164) and Gemella spp. (n = 28). The macrolide resistance phenotype was predominant (59.38%); all isolates with this phenotype carried the mef(A) or mef(E) gene, with mef(E) being predominant (95.36%). The erm(B) gene was always detected in strains with constitutive and inducible MLS(B) resistance and was combined with the mef(A/E) gene in 47.44% of isolates. None of the isolates carried the erm(A) subclass erm(TR), erm(A), or erm(C) genes. The mel gene was detected in all but four strains carrying the mef(A/E) gene. The tet(M) gene was found in 86.90% of tetracycline-resistant isolates and was strongly associated with the presence of the erm(B) gene. The cat(pC194) gene was detected in seven chloramphenicol-resistant Streptococcus mitis isolates, and the aph(3')-III gene was detected in four viridans group streptococcal isolates with high-level kanamycin resistance. The intTn gene was found in all isolates with the erm(B), tet(M), aph(3')-III, and cat(pC194) gene. The mef(E) and mel genes were successfully transferred from both groups of bacteria to Streptococcus pneumoniae R6 by transformation. Viridans group streptococci and Gemella spp. seem to be important reservoirs of resistance genes.