Beta-lactam antibiotics and viridans group streptococci / Antibióticos beta-lactámicos y estreptococos grupo viridans

Abstract The aim of this review is to present an update on the susceptibility of viridans groupstreptococci (VGS) to -lactam antimicrobials, with emphasis on the Argentinean scenario. VGSare a heterogeneous group including five groups of species, each one exhibiting peculiar sus-ceptibility patterns to penicillin (PEN). Species of the Streptococcus mitis group are frequentlynonsusceptible to PEN. PEN resistance is associated with changes in PEN-binding proteins. InArgentina, one to two thirds of VGS are nonsusceptible to PEN. Third generation cephalosporinsand carbapenems are currently more effective in vitro than PEN against VGS. Mortality was asso-ciated to nonsusceptibility to PEN in at least two studies involving patients with bacteremiacaused by VGS. Treatment of endocarditis due to VGS should be adjusted/to the PEN suscepti-bility of the isolates. Vancomycin may be an alternative choice for treating endocarditis causedby PEN-resistant isolates (MIC 4 g/ml).

Resumen El objetivo de esta revisión es presentar una actualización sobre la sensibilidad de los estreptococos del grupo viridans (EGV) a los antimicrobianos p-lactámicos, con énfasis en el escenario argentino. Los EGV son un grupo heterogéneo que incluye cinco grupos de especies, y cada una presenta su patrón especial de sensibilidad a la penicilina (PEN). Las especies del grupo Streptococcus mitis, con mayor frecuencia, no son sensibles a la PEN. La resistencia a la PEN se asocia con cambios de las proteínas ligadoras de PEN. En la Argentina, de uno a dos tercios de los EGV no son sensibles a la PEN. Las cefalosporinas de tercera generación y los carbapenemes son actualmente más eficaces in vitro que la PEN contra los EGV. La mortalidad se asoció con la no sensibilidad a la PEN en al menos dos estudios de pacientes con bacteriemia por EGV. El tratamiento de las endocarditis por EGV debe ajustarse según la sensibilidad a la PEN de los aislados. La vancomicina podría ser una elección alternativa para el tratamiento de las endocarditis por cepas resistentes a PEN (CIM >4 ^g/ml).

Beta-lactam antibiotics and viridans group streptococci.

The aim of this review is to present an update on the susceptibility of viridans group streptococci (VGS) to ß-lactam antimicrobials, with emphasis on the Argentinean scenario. VGS are a heterogeneous group including five groups of species, each one exhibiting peculiar susceptibility patterns to penicillin (PEN). Species of the Streptococcus mitis group are frequently nonsusceptible to PEN. PEN resistance is associated with changes in PEN-binding proteins. In Argentina, one to two thirds of VGS are nonsusceptible to PEN. Third generation cephalosporins and carbapenems are currently more effective in vitro than PEN against VGS. Mortality was associated to nonsusceptibility to PEN in at least two studies involving patients with bacteremia caused by VGS. Treatment of endocarditis due to VGS should be adjusted/to the PEN susceptibility of the isolates. Vancomycin may be an alternative choice for treating endocarditis caused by PEN-resistant isolates (MIC≥4µg/ml).

Epidemiology of Invasive Infections Caused by Streptococcus agalactiae in Argentina.

In recent years, an increase in the number of cases of invasive infections due to Streptococcus agalactiae (group B Streptococcus [GBS]) in adults has been reported. During 2014 and 2015, a multicentric, observational, and prospective study, including 40 health centers, was developed to describe the phenotypic and genotypic characteristics of GBS isolates circulating in Argentina and to analyze the clinical and demographic characteristics of patients with invasive infections. We recovered 162 invasive (GBSi) isolates from adult patients (n = 130, 80.2%), neonates (n = 24, 15%), and children below 18 years of age (n = 8, 4.8%), and colonizing (GBSc) isolates were recovered in prenatal GBS screening. GBS infection in adults was associated with underlying diseases, mainly diabetes mellitus. All isolates were penicillin susceptible. Resistance rates to erythromycin (25%) and clindamycin (26%) among the GBSc isolates were significantly higher than those from GBSi strains (17.3% and 16.1%, respectively); by contrast, levofloxacin resistance was significantly higher in GBSi isolates (14.8% vs. 7%). Serotype Ia was the most frequent in neonates and Ib was most frequent in adults. Serotypes Ia and III were prevalent in GBSc isolates. The increase of levofloxacin resistance was associated with the presence of a serotype Ib clone. This work emphasizes the need for GBS infection surveillance studies to implement correct treatments and adequate prevention strategies.

Resistance to ß-lactams in Streptococcus pneumoniae.

Streptococcus pneumoniae is an important causal agent of pneumonia, meningitis, sepsis, bacteremia, and otitis media. Penicillin resistance rates in S. pneumoniae have remained stable in Argentina in the last years. In the late '90s more isolates with MIC of penicillin ≥2µg/ml were observed; however, their frequency has decreased in recent years. The phenotypic expression of penicillin resistance is due to a modification in penicillin-binding proteins associated with a mosaic structure in the coding genes. The expansion of successful resistant clones varies among the different regions and is influenced by the use of antibiotics, vaccines, particularly conjugated ones, as well as population density. Parenteral treatment with high doses of penicillin G continues to be effective for the treatment of pneumonia and bacteremia, oral aminopenicillins for otitis media and sinusitis and third generation cephalosporins for meningitis.

Resistencia a los β -lactámicos en enterococos / Resistance to β-lactams in enterococci

Enterococci are intrinsically resistant to several antimicrobial classes and show a great ability to acquire new mechanisms of resistance. Resistance to β-lactam antibiotics is a major concern because these drugs either alone or in combination are commonly used for the treatment of enterococcal infections. Ampicillin resistance, which is rare in Enterococcus faecium occurs in most of the hospital-associated Enterococcus faecium isolates. High-level resistance to ampicillin in E. faecium is mainly due to the enhanced production of PBP5 and/or by polymorphisms in the beta subunit of this protein. The dissemination of high-level ampicillin resistance can be the result of both clonal spread of strains with mutated pbp5 genes and resistance horizontal gene transfer.

Los enterococos son intrínsecamente resistentes a varias clases de antimicrobianos y presentan una gran capacidad para adquirir mecanismos de resistencia. La resistencia a los antibióticos p-lactámicos es preocupante porque estos fármacos solos o combinados se usan comúnmente para el tratamiento de las infecciones enterocócicas. La mayoría de los aislamientos hospitalarios de Enterococcus faecium presentan resistencia a la ampicilina, la cual es rara en Enterococcus faecalis. El alto nivel de resistencia a la ampicilina en E. faecium se debe principalmente a la hiperproducción de PBP5 y/o a polimorfismos en la subunidad beta de esta proteína. La propagación de esta resistencia puede deberse tanto a la diseminación clonal de cepas con genes pbp5 mutados como a la transferencia horizontal de genes.

Identificación y sensibilidad a los antimicrobianos de aislados de estreptococos del grupo viridans provenientes de pacientes internados en un hospital universitario de la ciudad de Buenos Aires / Identification and antibiotic susceptibility of viridans group streptococci isolates recovered from patients hospitalized at a teaching hospital in Buenos Aires City

Los estreptococos del grupo viridans (EGV) son agentes causales de infecciones localizadas e invasivas. Dada la gravedad de las infecciones producidas por EGV sumada a las escasas comunicaciones actuales en nuestro país, los objetivos de este trabajo fueron la identificación y el estudio de la sensibilidad a los antibióticos de aislados caracterizados como EGV, recuperados de pacientes internados, para actualizar el conocimiento sobre el perfil de resistencia y la epidemiología de las infecciones ocasionadas por EGV. Se recuperaron 132 aislados de EGV en el Hospital de Clínicas «José de San Martín¼ en el período 2011-2015. La identificación se realizó mediante pruebas convencionales y espectrometría de masas (Matrix Assisted Laser Desorption Ionization - Time of Flight Mass Spectrometry). El grupo Streptococcus anginosus fue el más frecuente (42%) seguido por el grupo Streptococcus mitis (33%). Dentro del grupo S. mitis se excluyó a Streptococcus pneumoniae. El 100% de los aislados fue sensible a ertapenem, linezolid y vancomicina; el 96,9% a cef-triaxona y cefepima. Se encontró un 25,8% de resistencia a penicilina (I+R) fundamentalmente en aislados de grupo S. mitis. La resistencia a tetraciclina fue del 27,2% y solo 2/132 aislados fueron resistentes a levofloxacina. Los valores de CIM de gentamicina oscilaron entre 0,5 y 32 -og/ml. El 17,4% de los aislados presentó resistencia a eritromicina sin diferencia significativa en la distribución de fenotipos M y MLS. Los resultados muestran la importancia de la vigilancia continua de las infecciones producidas por estos microorganismos con el fin de generar aportes para la elección de la terapia antibiótica adecuada.

Members of the viridans group streptococci (VGS) are the cause of local and invasive infections. Due to the severity of these infections and taking into account that reports regarding epidemiological aspects are scarce, the aims of this work were the identification and the study of the antibiotic susceptibility profiles of the isolates recovered from patients that were hospitalized in order to find out about the resistance level and the epidemiology of infections in which VGS are involved. A hundred and thirty two isolates identified as VGS were isolated at Hospital de Clínicas «José de San Martín¼ during the period 2011-2015. The identification was performed by biochemical test and mass spectrometry by Matrix Assisted Laser Desorption Ionization -Time of Flight Mass Spectrometry. Streptococcus anginosus group was prevalent (42%) followed by Streptococcus mitis group (33%). In the latter, isolates of Streptococcus pneumoniae were excluded. All the VGS isolates were susceptible to ertapenem, meropenem, linezolid and vancomycin; 25.8% were resistant (I+R) to penicillin, being prevalent in the S. mitis group. Regarding ceftriaxone and cefepime 96.9% of the isolates were susceptible. Only two isolates were resistant to levofloxacin, 27.2% to tetracycline and it was not found high level resistance to gentamycin (MIC range 0.5-32 µg/ml). Resistance to erythromycin was 17.4% with no significant difference between M and MLS phenotypes. The most active antibiotics were in addition to ceftriaxone and cefepime, vancomycin, ertapenem, meropenem and linezolid. These results highlight the importance of the continuous surveillance of the infections caused by VGS in order to predict a correct antibiotic therapy.

Fluoroquinolone-Resistant Streptococcus agalactiae Invasive Isolates Recovered in Argentina.

Background: Streptococcus agalactiae or group B Streptococcus (GBS) is an important pathogen in neonates and nonpregnant individuals. Epidemiological studies of GBS resistance to fluoroquinolones (FQs) in Latin America are scarce. This study aimed to determine the local prevalence of FQ resistance in the frame of a national, prospective multicenter study of invasive GBS infections and to investigate mechanisms of resistance, serotype distribution, and clonal relationships among resistant isolates. Methods: From July 2014 to July 2015, 162 invasive GBS isolates were collected from 86 health care centers in 32 Argentinean cities. All isolates were screened for FQ nonsusceptibility using a five-disc scheme: levofloxacin (LVX), ciprofloxacin, norfloxacin (NOR), ofloxacin, and pefloxacin (PF). LVX minimal inhibitory concentration (MIC) was determined by the agar dilution method. Sequencing of internal regions of gyrA and parC genes was performed. Capsular typing and genetic characterization of nonsusceptible isolates were assessed by latex agglutination, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing. Results: Twenty-four of one hundred sixty-two GBS isolates exhibited no inhibition zones to all tested FQs with an MIC range of 16-32 mg/L for LVX, and one isolate with MIC = 1 mg/L showed no inhibition zones around NOR and PF discs. In all resistant isolates, point mutations were detected in both genes. Serotype Ib was prevalent (88%). One PFGE type accounted for 84% of the FQ-resistant isolates and belonged to serotype Ib, sequence type 10. Conclusions: The prevalence of FQ resistance was 14.8% likely to be associated with dissemination of an ST10/serotype Ib clone. The unexpected high rate of resistance emphasizes the relevance for continuous surveillance of GBS epidemiology and antibiotic susceptibility.

Argentinian multicenter study on urinary tract infections due to Streptococcus agalactiae in adult patients.

INTRODUCTION: Streptococcus agalactiae (group B streptococcus, GBS) is a recognized urinary pathogen both in males and pregnant or non-pregnant women. Data regarding GBS serotypes recovered from urinary tract infections (UTIs) are scarce. The aim of this study was to describe the clinical and microbiological characteristics of UTIs caused by GBS in adult patients in Argentina. METHODOLOGY: A prospective multicenter study involving 86 centers was conducted in Argentina between July 1st, 2014 and June 30th, 2015. Antimicrobial susceptibility and serotype distribution of GBS isolated from the urinary tract of adult patients were determined. Susceptibility tests were performed by the disk diffusion and/or agar dilution methods. Epidemiological and clinical characteristics of the patients were considered to identify associated comorbilities. RESULTS: Seven hundred and one GBS were sent to the reference laboratory in the above mentioned period, however, only 211 fulfilled our selection criteria (demographic data availability, underlying diseases reported, colony counts greater than 105 CFU/mL, single organism isolated from the urine sample). No penicillin-resistant GBS was found but fluoroquinolone resistance was high (12.8%), especially among GBS isolated from men and non-pregnant women. UTIs due to GBS were associated to underlying diseases in men and non-pregnant women, particularly diabetes mellitus. Most of the isolates showed serotypes Ia and III. CONCLUSIONS: GBS are still susceptible to penicillin but fluoroquinolone resistance is a growing concern, at least in Argentina. There are underlying conditions that could be associated to urinary infections caused by GBS.

Resistance to ß-lactams in enterococci.

Enterococci are intrinsically resistant to several antimicrobial classes and show a great ability to acquire new mechanisms of resistance. Resistance to ß-lactam antibiotics is a major concern because these drugs either alone or in combination are commonly used for the treatment of enterococcal infections. Ampicillin resistance, which is rare in Enterococcus faecalis, occurs in most of the hospital-associated Enterococcus faecium isolates. High-level resistance to ampicillin in E. faecium is mainly due to the enhanced production of PBP5 and/or by polymorphisms in the beta subunit of this protein. The dissemination of high-level ampicillin resistance can be the result of both clonal spread of strains with mutated pbp5 genes and horizontal gene transfer.

[Identification and antibiotic susceptibility of viridans group streptococci isolates recovered from patients hospitalized at a teaching hospital in Buenos Aires City]. / Identificación y sensibilidad a los antimicrobianos de aislados de estreptococos del grupo viridans provenientes de pacientes internados en un hospital universitario de la ciudad de Buenos Aires.

Members of the viridans group streptococci (VGS) are the cause of local and invasive infections. Due to the severity of these infections and taking into account that reports regarding epidemiological aspects are scarce, the aims of this work were the identification and the study of the antibiotic susceptibility profiles of the isolates recovered from patients that were hospitalized in order to find out about the resistance level and the epidemiology of infections in which VGS are involved. A hundred and thirty two isolates identified as VGS were isolated at Hospital de Clínicas «José de San Martín¼ during the period 2011-2015. The identification was performed by biochemical test and mass spectrometry by Matrix Assisted Laser Desorption Ionization -Time of Flight Mass Spectrometry. Streptococcus anginosus group was prevalent (42%) followed by Streptococcus mitis group (33%). In the latter, isolates of Streptococcus pneumoniae were excluded. All the VGS isolates were susceptible to ertapenem, meropenem, linezolid and vancomycin; 25.8% were resistant (I+R) to penicillin, being prevalent in the S. mitis group. Regarding ceftriaxone and cefepime 96.9% of the isolates were susceptible. Only two isolates were resistant to levofloxacin, 27.2% to tetracycline and it was not found high level resistance to gentamycin (MIC range 0.5-32µg/ml). Resistance to erythromycin was 17.4% with no significant difference between M and MLS phenotypes. The most active antibiotics were in addition to ceftriaxone and cefepime, vancomycin, ertapenem, meropenem and linezolid. These results highlight the importance of the continuous surveillance of the infections caused by VGS in order to predict a correct antibiotic therapy.

Susceptibility to β-lactams in β-hemolytic streptococci / Sensibilidad a los β-lactámicos en estreptococos β-hemolíticos

Group A (GAS), B (GBS), c (GCS) and G (GGS) β-hemolytic streptococci are important human pathogens. They cause infections of different severity and frequency. Nowadays, after 70 years of use, penicillin is still universally active against GAS, GCS and GGS. However, therapeutic failures have been recorded in 2-28% of pharyngitis cases (median: 12%) attributable to different causes. By contrast, some GBS with reduced susceptibility to penicillin have been described, especially in Japan. In this group of bacteria, it is important to highlight that confirmation by reference methods is mandatory when decreased susceptibility to penicillin is suspected as well as checked for the detection of the mechanisms involved.

Los estreptococos β-hemolíticos de los grupos A (GAS), B (GBS), C (GCS) y G (GGS) son importantes patógenos humanos. Ellos producen infecciones de diversa gravedad y frecuencia. Aún después de más de 70 años de uso, la penicilina sigue siendo activa in vitro frente al 100% de los GAS, GCS y GGS. No obstante se han producido fallas terapéuticas entre el 2-28% de los casos de faringitis (media: 12%), atribuibles a diversas causas. En cambio se han descrito aislados de GBS con sensibilidad reducida a la penicilina, especialmente en Japón. Es importante que toda sospecha de sensibilidad disminuida a la penicilina en este grupo de bacterias sea confirmada por los métodos de referencia y comprobada mediante la detección de los mecanismos involucrados.

[Susceptibility of Streptococcus pyogenes isolated from invasive infections to trimethoprim-sulfamethoxazole]. / Sensibilidad a trimetoprima-sulfametoxazol de Streptococcus pyogenes aislados de infecciones invasivas.

It is erroneously believed that group A streptococci (GAS) are universally resistant to trimethoprim-sulfamethoxazole (TMS). This is mainly because media commonly used for in vitro determination of susceptibility to antibiotics contain thymidine, a nucleoside that antagonizes the antibiotic effect of TMS. The objective of this work was to determine EGA sensitivity to TMS in the presence and absence of thymidine. To this aim, 95 GAS isolates obtained from clinical tissues with i nvasive infections were analyzed. Susceptibility tests were performed by diffusion with TMS discs in Mueller Hinton agar supplemented either with 5% sheep blood or with 5% lysed equine blood (MH-LEB). Lysed equine blood contains thymidine phosphorylase, which degrades this nucleoside. Epsilometry (Etest) was used as gold standard. Quality controls with Enterococcus faecalis strain ATCC 29212 were satisfactory with both media. A 100% sensitivity to TMS was found in MH-SEL whereas 6 isolates (6.3%) resulted resistant in MH-SC; only one of them was found to have intermediate susceptibility by Etest (MIC > 1.5/28 υg/ml). The genetic determinants most frequently associated to TMS resistant EGA were not found in this isolate. Probably, if more accurate GAS-specific cut-off points were established for diffusion, the correlation with dilution methods or with the Etest could be improved, even employing MH-SB.

Sensibilidad a trimetoprima-sulfametoxazol de Streptococcus pyogenes aislados de infecciones invasivas / Susceptibility of Streptococcus pyogenes isolated from invasive infections to trimethoprim-sulfamethoxazole

Se cree erróneamente que los estreptococos del grupo A (EGA) son universalmente resistentes a trimetoprima-sulfametoxazol (TMS). Esto se debe a que la timidina presente en los medios habitualmente usados para determinar sensibilidad in vitro a antibióticos antagoniza el efecto antibiótico de TMS. El objetivo de este trabajo fue determinar la sensibilidad de EGA a TMS, en presencia y ausencia de timidina. A tal fin, fueron analizados 95 aislamientos clínicos obtenidos de tejidos normalmente estériles con infección invasiva por EGA. La pruebas de sensibilidad por difusión con discos de TMS fueron realizadas en agar Mueller Hinton adicionado ya sea con 5% de sangre de carnero (MH-SC) o con 5% de sangre equina lisada (MH-SEL). La sangre equina lisada contiene timidina fosforilasa, que degrada este nucleósido. Como método de referencia se utilizó la epsilometría (Etest). El control de calidad con la cepa Enterococcus faecalis ATCC 29212 fue satisfactorio para ambos medios. La sensibilidad a TMS por difusión fue 100% en MH-SEL; en agar MH-SC 6 (6.3%) aislamientos resultaron resistentes; por Etest todos fueron sensibles, excepto uno de esos seis que presentó sensibilidad intermedia (CIM = 1.5/28.5 μg/ml). En este aislamiento no se encontraron las mutaciones genéticas de EGA más frecuentemente asociadas a resistencia a TMS. Probablemente, si se establecieran mejores puntos de corte para difusión, específicos para EGA, podría optimizarse la correlación con métodos de dilución o con Etest, aun empleando MH-SC.

It is erroneously believed that group A streptococci (GAS) are universally resistant to trimethoprim-sulfamethoxazole (TMS). This is mainly because media commonly used for in vitro determination of susceptibility to antibiotics contain thymidine, a nucleoside that antagonizes the antibiotic effect of TMS. The objective of this work was to determine EGA sensitivity to TMS in the presence and absence of thymidine. To this aim, 95 GAS isolates obtained from clinical tissues with i nvasive infections were analyzed. Susceptibility tests were performed by diffusion with TMS discs in Mueller Hinton agar supplemented either with 5% sheep blood or with 5% lysed equine blood (MH-LEB). Lysed equine blood contains thymidine phosphorylase, which degrades this nucleoside. Epsilometry (Etest) was used as gold standard. Quality controls with Enterococcus faecalis strain ATCC 29212 were satisfactory with both media. A 100% sensitivity to TMS was found in MH-SEL whereas 6 isolates (6.3%) resulted resistant in MH-SC; only one of them was found to have intermediate susceptibility by Etest (MIC > 1.5/28 μg/ml). The genetic determinants most frequently associated to TMS resistant EGA were not found in this isolate. Probably, if more accurate GAS-specific cut-off points were established for diffusion, the correlation with dilution methods or with the Etest could be improved, even employing MH-SB.

Susceptibility to ß-lactams in ß-hemolytic streptococci.

Group A (GAS), B (GBS), C (GCS) and G (GGS) ß-hemolytic streptococci are important human pathogens. They cause infections of different severity and frequency. Nowadays, after 70 years of use, penicillin is still universally active against GAS, GCS and GGS. However, therapeutic failures have been recorded in 2-28% of pharyngitis cases (median: 12%) attributable to different causes. By contrast, some GBS with reduced susceptibility to penicillin have been described, especially in Japan. In this group of bacteria, it is important to highlight that confirmation by reference methods is mandatory when decreased susceptibility to penicillin is suspected as well as checked for the detection of the mechanisms involved.

Group B streptococcal necrotizing pneumonia in a diabetic adult patient / Neumonía necrosante por estreptococo del grupo B en un paciente adulto diabético

The aim of this report is to describe a rare case of necrotizing pneumonia due to group B Streptococcus serotype III in a relatively young male adult (48 years old) suffering from diabetes. The organism was isolated from his pleural fluid and was only resistant to tetracycline. The patient first received ceftazidime (2 g/8 h i.v.) + clindamycin (300 mg/8 h) for 18 days and then he was discharged home and orally treated with amoxicillin clavulanic acid (1 g/12 h) for 23 days with an uneventful evolution. As in the cases of invasive infection by Streptococcus pyogenes, clindamycin could prevent streptococcal toxic shock syndrome.

El objetivo de esta presentación es describir un caso raro de neumonía necrosante debida a estreptococo del grupo B serotipo III en un diabético adulto de sexo masculino relativamente joven (48 años). El microorganismo fue aislado de líquido pleural y resultó ser resistente solo a tetraciclina. El paciente recibió ceftacidima (2 g/8 h iv) + clindamicina (300 mg/8 h) durante 18 días y luego fue dado de alta, bajo tratamiento oral con amoxicilina-ácido clavulánico (1 g/12 h). Este tratamiento se mantuvo durante 23 días, con buena evolución. Como en casos de infecciones invasivas por Streptococcus pyogenes, es posible que la clindamicina haya evitado la aparición del síndrome de shock tóxico estreptocócico.

Group B streptococcal necrotizing pneumonia in a diabetic adult patient.

The aim of this report is to describe a rare case of necrotizing pneumonia due to group B Streptococcus serotype III in a relatively young male adult (48 years old) suffering from diabetes. The organism was isolated from his pleural fluid and was only resistant to tetracycline. The patient first received ceftazidime (2g/8h i.v.)+clindamycin (300mg/8h) for 18 days and then he was discharged home and orally treated with amoxicillin clavulanic acid (1g/12h) for 23 days with an uneventful evolution. As in the cases of invasive infection by Streptococcus pyogenes, clindamycin could prevent streptococcal toxic shock syndrome.

Screening assay for inhibitors of a recombinant Streptococcus pneumoniae UDP-glucose pyrophosphorylase.

The UDP-glucose pyrophosphorylase of Streptococcus pneumoniae (GalUSpn) is absolutely required for the biosynthesis of capsular polysaccharide, the sine qua non virulence factor of pneumococcus. Since the eukaryotic enzymes are completely unrelated to their prokaryotic counterparts, we propose that the GalU enzyme is a critical target to fight the pneumococcal disease. A recombinant GalUSpn was overexpressed and purified. An enzymatic assay that is rapid, sensitive and easy to perform was developed. This assay was appropriate for screening chemical libraries for searching GalU inhibitors. This work represents a fundamental step in the exploration of novel antipneumococcal drugs.

Antimicrobial susceptibility of clinical isolates of Actinomyces and related genera reveals an unusual clindamycin resistance among Actinomyces urogenitalis strains.

OBJECTIVES: Patterns of antimicrobial susceptibility in Actinomyces and related genera are very limited in the literature. Data of predominant susceptibility profiles could contribute to the establishment of an accurate empirical treatment. METHODS: A total of 113 isolates from clinical samples were included in this study. Each isolate was identified using phenotypic methods and MALDI-TOF/MS. When discrepancies were observed, 16S rRNA gene sequencing was performed. The minimum inhibitory concentrations (MICs) of nine antimicrobial agents (penicillin, ceftriaxone, linezolid, tetracycline, clindamycin, erythromycin, ciprofloxacin, levofloxacin and vancomycin) were tested against the species Actinotignum schaalii (n=23), Actinomyces turicensis (n=18), Actinomyces europaeus (n=13), Actinomyces naeslundii/Actinomyces viscosus group (n=12), Actinomyces urogenitalis (n=11), Actinomyces radingae (n=11), Actinomyces neuii (n=9), Actinomyces odontolyticus (n=8), Bifidobacterium scardovii (n=3), Actinomyces graevenitzii (n=2), Alloscardovia omnicolens (n=2) and Varibaculum cambriense (n=1). RESULTS: All of the isolates were susceptible to penicillin, ceftriaxone, vancomycin and linezolid. Almost all of the A. urogenitalis isolates (8/11) were resistant to clindamycin and showed susceptibility to erythromycin, suggesting an L-phenotype, however no determinants of clindamycin resistance (lnu and lsa genes) were detected by PCR. High MIC values to quinolones were observed in 54/113 isolates (47.8%). All of the A. urogenitalis isolates were highly resistant to ciprofloxacin and levofloxacin. CONCLUSIONS: These data highlight the importance of ongoing surveillance to provide relevant information for empirical management of infections caused by these organisms.

The galU gene expression in Streptococcus pneumoniae.

The polysaccharide capsule of Streptococcus pneumoniae is the main virulence factor making the bacterium resistant to phagocytosis. The galU gene of S. pneumoniae encodes a UDP-glucose pyrophosphorylase absolutely required for capsule biosynthesis. In silico analyses indicated that the galU gene is co-transcribed with the gpdA gene, and four putative promoter regions located upstream of gpdA were predicted. One of them behaved as a functional promoter in a promoter reporter system. It is conceivable that the sequence responsible for initiating transcription of gpdA-galU operon is an extended -10 site TATGATA(T/G)AAT. Semi-quantitative real-time reverse transcription PCR experiments indicated that galU was expressed mainly in the exponential phase of growth.