Susceptibilidad antimicrobiana en muestras urogenitales de ureaplasma urealyticum, el principal agente en infección intramaniótica / Antimicrobial susceptibility in urogenital samples of ureaplasma urealyticum, the main agent in intramaniotic infection

RESUMEN OBJETIVO: Ureaplasma urealyticum es el agente más frecuentemente aislado en infección intraamniótica. Los macrólidos son los antimicrobianos de primera elección en embarazadas. Se ha descrito el aumento de resistencia, pudiendo limitar las opciones terapéuticas durante la gestación. El propósito del estudio es evaluar susceptibilidad antimicrobiana de Ureaplasma urealyticum aislado en mujeres en edad fértil, que se atienden en Clínica Alemana Temuco, Araucanía, Chile. METODO: Se estudian todas las muestras de orina y flujo vaginal para cultivo de U. urealyticum, de pacientes entre 18 y 40 años, recibidas en el Laboratorio de Microbiología Clínica Alemana Temuco, en período Abril 2013 a Enero 2015. Se procesan las muestras con kit Mycoplasma IST 2 de Biomerieux. En las que resultan positivas, se estudia susceptibilidad a macrólidos, tetraciclinas y quinolonas. RESULTADOS: 426 muestras de orina y flujo vaginal (390 pacientes). 197 pacientes resultaron positivas para U. urealyticum. (50,5%). La susceptibilidad fue 88,4% (174 pctes) a Eritromicina, 87,9% (173 pctes) a Claritromicina y 91,9% (181 pctes) a Azitromicina (NS). 15 de 197 pacientes (7,6%) fueron resistentes a los 3 macrólidos. La susceptibilidad a Quinolonas fue 55,3% a Ciprofloxacino, y 94% a Ofloxacino. El 100% resultó susceptible a Tetraciclinas. CONCLUSIONES: Cerca del 10% de U. urealyticum aislados en nuestra serie son resistentes a macrólidos, contribuyendo a la no erradicación de la infección en tratamientos empíricos. Dentro de ellos, azitromicina aparece con la mayor efectividad. El aumento de resistencia limitará opciones terapéuticas, con gran impacto perinatal en futuro. La vigilancia de susceptibilidad en cada hospital es fundamental para elección terapéutica.

ABSTRACT INTRODUCTION: Ureaplasma urealyticum is the most frequently isolated microorganism in intra-amniotic infection. The macrolides are the first choice antimicrobials for treat this infection in pregnancy. The increasing resistance has been described worldwide, seriously limiting therapeutic options in pregnancy. The aim of the study is to evaluate antimicrobial susceptibility of U. urealyticum aislated in fertile-age women in Clínica Alemana Temuco, Araucania region, Chile. METHOD: Urine and vaginal samples were analyzed for U. urealyticum, from every 18 to 40 years old patients, received at Microbiology Laboratory of Clínica Alemana Temuco, between April 2013 to January 2015. The samples are processed with Mycoplasma IST 2 kit of Biomerieux. If they became positives, susceptibility to macrolides, tetracyclines and quinolones was studied. RESULTS: 426 urine and vaginal samples were collected (390 patients). 197 patients were positive for U. urealyticum (50.5%). The susceptibility was 88.4% (174 pts) to Erythromicyn, 87.9% (173 pts) to Clarithromycin and 91.9% (181 pts) to Azithromycin (NS). Resistance to all macrolides was observed in 15 out of 197 patients (7.6%). The susceptibility to Quinolones was 55.3% to Ciprofloxacin, and 94% to Ofloxacin. The 100% was susceptible to Tetracyclines. DISCUSSION: Near to 10% of isolated Ureaplasma spp in our serie were resistant to some macrolide, being a factor for failing to eradicate the infection in empirical treatment. Azithromycin was the most effective. The increasing resistance will limit therapeutic options, with great perinatal impact in the future. Susceptibility surveillance in each hospital is very important for therapeutic options.

Susceptibility of Streptococcus agalactiae to erythromycin and clindamycin in pregnant women in the prenatal screening

Introduction: Group B streptococcus (GBS), or Streptococcus agalactiae, is a bacterium found in normal human microbiota. However, it may cause neonatal pneumonia, sepsis, and meningitis. Genital colonization in pregnant women is associated with a higher risk of preterm birth. The treatment of choice is antibiotic therapy with beta-lactams, but in the case of multidrug-resistance, erythromycin and clindamycin can be used. Methods: This study evaluated bacterial cultures in the period from 2014 to 2015 from a group of 29,875 pregnant women. GBS colonization and resistance to erythromycin and clindamycin were investigated. Results: Positive cultures were found in 26.8% and 26.1% of the samples in 2014 and 2015, respectively. Levels of resistance to erythromycin and clindamycin were, respectively, 2.4% and 5.5% in 2014 and 3.2% and 6.5% in 2015. Conclusion: The investigation of GBS colonization and the evaluation of GBS resistance to erythromycin and clindamycin are of extreme relevance, given the increasing incidence of bacterial resistance, risks of preterm birth. (AU)

The prevalence of genotypes that determine resistance to macrolides, lincosamides, and streptogramins B compared with spiramycin susceptibility among erythromycin-resistant Staphylococcus epidermidis

Coagulase-negative staphylococci, particularly Staphylococcus epidermidis, can be regarded as potential reservoirs of resistance genes for pathogenic strains, e.g., Staphylococcus aureus. The aim of this study was to assess the prevalence of different resistance phenotypes to macrolide, lincosamide, and streptogramins B (MLSB) antibiotics among erythromycin-resistant S. epidermidis, together with the evaluation of genes promoting the following different types of MLSB resistance:ermA, ermB, ermC,msrA, mphC, and linA/A’. Susceptibility to spiramycin was also examined. Among 75 erythromycin-resistantS. epidermidis isolates, the most frequent phenotypes were macrolides and streptogramins B (MSB) and constitutive MLSB (cMLSB). Moreover, all strains with the cMLSB phenotype and the majority of inducible MLSB (iMLSB) isolates were resistant to spiramycin, whereas strains with the MSB phenotype were sensitive to this antibiotic. The D-shape zone of inhibition around the clindamycin disc near the spiramycin disc was found for some spiramycin-resistant strains with the iMLSB phenotype, suggesting an induction of resistance to clindamycin by this 16-membered macrolide. The most frequently isolated gene was ermC, irrespective of the MLSB resistance phenotype, whereas the most often noted gene combination wasermC, mphC, linA/A’. The results obtained showed that the genes responsible for different mechanisms of MLSB resistance in S. epidermidis generally coexist, often without the phenotypic expression of each of them.

Epidemiological Study of Erythromycin-Resistant Streptococcus pyogenes From Korea and Japan by emm Genotyping and Multilocus Sequence Typing

BACKGROUND: We determined the epidemiological characteristics of erythromycin (EM)-resistant Streptococcus pyogenes (group A streptococci, GAS) strains isolated from Korea and Japan, using emm genotyping and multilocus sequence typing (MLST). METHODS: Clinical isolates of GAS had been collected from 1992 to 2012 in Korea and from 2004 to 2009 in Japan. EM resistance was determined by the microdilution method, and resistance genotypes were assessed by PCR. The emm genotyping and MLST were performed by DNA sequencing. RESULTS: The emm genotypes and sequence types (STs) were concordant in 143 (85.1%) of 168 EM-resistant GAS strains from Korea. ST36/emm12 (35.1%), ST52/emm28 (22.6%), and ST49/emm75 (16.1%) were the most common types. Most of the ST36 (93.9%) and ST52 (95.8%) strains harbored erm(B), whereas strains ST49, ST42, and ST15 contained mef(A). The concordance between emm genotypes and STs was 41 (93.2%) among 44 EM-resistant GAS strains from Japan. ST36/emm12 (34.1%), ST49/emm75 (18.2%), and ST28/emm1 (15.9%) were the major types. ST36 isolates harbored either erm(B) (56.3%) or mef(A) (37.5%), whereas isolates ST28, ST49, and ST38 carried only mef(A). The proportion of erm(B) and mef(A) was 66.1% and 33.3% in Korea and 22.7% and 68.2% in Japan, respectively. CONCLUSIONS: The common STs in Korea and Japan were ST36 and ST49, whereas ST52 was present only in Korea and ST28 only in Japan. Genotype erm(B) was predominant in Korea, whereas mef(A) was frequent in Japan. There were differences between Korea and Japan regarding the frequencies of emm genotypes, STs, and EM resistance genes among the EM-resistant GAS.

Inducible clindamycin resistance in Staphylococcus aureus: Reason for treatment failure.

Emergence of methicillin resistance in Staphylococcus aureus (S.aureus) has left us with very few therapeutic alternatives available to treat staphylococcal infection. The widespread use of macrolide-lincosamide-streptogramin B (MLSB) antibiotics has led to an increase in number of staphylococcal strains acquiring resistance to MLSB antibiotics. This study was done to investigate the infections by hospital and community acquired “erythromycin- induced clindamycin resistant” strains or D-test positives of clinical isolates of Staphylococcus aureus (S.aureus) in a hospital. Three hundred isolates of S.aureus were subjected to routine antibiotic susceptibility testing including Cefoxitin (30µg) by modified disc diffusion method. Inducible resistance to clindamycin in S.aureus was tested by D-test as per CLSI guidelines. Among 300 S.aureus isolates, 114 (38%) were methicillin resistant Staphylococcus aureus (MRSA) and 186 (62%) methicillin susceptible Staphylococcus aureus (MSSA). Forty one (13.67%) isolates showed induced clindamycin resistance, 49(16.33%) showed constitutive resistance and 94 (31.33%) showed the MS phenotype. Inducible resistance and constitutive resistance were found to be higher in MRSA compared to MSSA (22.81%, 23.68% and 8.1%, 11.8% respectively). D-test should be included as a part of routine antibiotic susceptibility testing to detect induced clindamycin resistance to prevent treatment failure.

Effects of penicillin and erythromycin on adherence of invasive and noninvasive isolates of Streptococcus pyogenes to laminin

This study investigated the possible relationship between the invasiveness of group A Streptococcus (GAS) strains and their abilities to adhere to laminin and assessed the effects of subinhibitory concentrations of penicillin and erythromycin on the ability of GAS to adhere to laminin. The adherence of noninvasive and highly invasive isolates of GAS to laminin was significantly higher than the adherence displayed by isolates of low invasiveness. Antibiotic treatment caused significant reductions in adherence to laminin in all three groups of strains. Penicillin was more successful in reducing the adherence abilities of the tested GAS strains than erythromycin.

Detection of the Efflux-Mediated Erythromycin Resistance Transposon in Streptococcus pneumoniae

BACKGROUND: The present analysis focuses on phenotypic and genotypic characterizations of efflux-mediated erythromycin resistance in Streptococcus pneumoniae due to an increase in macrolide resistance in S. pneumoniae worldwide. METHODS: We investigated the prevalence of efflux-mediated erythromycin resistance and its relevant genetic elements from 186 specimens of S. pneumonia isolated from clinical and normal flora from Tehran, Iran. The presence of erythromycin resistance genes was tested by PCR with two sets of primers, specific for erm(B) and mef(A/E), and their genetic elements with tetM, xis, and int genes. Isolates were typed with the BOX PCR method and tested for resistance to six antibiotics. RESULTS: Antibiotic susceptibility tests revealed that 100% and 47% isolates were resistant to tetracycline and erythromycin, respectively. The erythromycin and clindamycin double-disc diffusion test for macrolide-lincosamide-streptograminB (MLSB) resistance phenotype showed 74 (84%) isolates with the constitutive MLSB phenotype and the remaining with the M phenotype. BOX PCR demonstrated the presence of 7 types in pneumococci with the M phenotype. Fourteen (16%) isolates with the M phenotype harbored mef(A/E), tetM, xis, and int genes. CONCLUSIONS: The present results suggest dissemination of polyclonal groups of S. pneumoniae with the M phenotype carrying resistance genes attributed to transposon 2009.

Antimicrobial susceptibility patterns, emm type distribution and genetic diversity of Streptococcus pyogenes recovered in Brazil

Streptococcus pyogenes is responsible for a variety of infectious diseases and immunological complications. In this study, 91 isolates of S. pyogenes recovered from oropharynx secretions were submitted to antimicrobial susceptibility testing, emm typing and pulsed-field gel electrophoresis (PFGE) analysis. All isolates were susceptible to ceftriaxone, levofloxacin, penicillin G and vancomycin. Resistance to erythromycin and clindamycin was 15.4%, which is higher than previous reports from this area, while 20.9% of the isolates were not susceptible to tetracycline. The macrolide resistance phenotypes were cMLSB (10) and iMLSB (4). The ermB gene was predominant, followed by the ermA gene. Thirty-two emm types and subtypes were found, but five (emm1, emm4, emm12, emm22, emm81) were detected in 48% of the isolates. Three new emm subtypes were identified (emm1.74, emm58.14, emm76.7). There was a strong association between emm type and PFGE clustering. A variety of PFGE profiles as well as emm types were found among tetracycline and erythromycin-resistant isolates, demonstrating that antimicrobial resistant strains do not result from the expansion of one or a few clones. This study provides epidemiological data that contribute to the development of suitable strategies for the prevention and treatment of such infections in a poorly studied area.

Evaluation of constitutive and inducible resistance to clindamycin in clinical samples of Staphylococcus aureus from a tertiary hospital

Introduction Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have become common in hospitals and the community environment, and this wide resistance has limited patient treatment. Clindamycin (CL) represents an important alternative therapy for infections caused by S. aureus. Antimicrobial susceptibility testing using standard methods may not detect inducible CL resistance. This study was performed to detect the phenotypes of resistance to macrolides-lincosamides-streptogramin B (MLSB) antibiotics, including CL, in clinical samples of S. aureus from patients at a tertiary hospital in Santa Maria, State of Rio Grande do Sul, Brazil. Methods One hundred and forty clinical isolates were submitted to the disk diffusion induction test (D-test) with an erythromycin (ER) disk positioned at a distance of 20mm from a CL disk. The results were interpreted according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI). Results In this study, 29 (20.7%) of the 140 S. aureus samples were resistant to methicillin (MRSA), and 111 (79.3%) were susceptible to methicillin (MSSA). The constitutive resistance phenotype (cMLSB) was observed in 20 (14.3%) MRSA samples and in 5 (3.6%) MSSA samples, whereas the inducible resistance phenotype (iMLSB) was observed in 3 (2.1%) MRSA samples and in 8 (5.8%) MSSA samples. Conclusions The D-test is essential for detecting the iMLSB phenotype because the early identification of this phenotype allows clinicians to choose an appropriate treatment for patients. Furthermore, this test is simple, easy to perform and inexpensive. .

Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae

Streptococcus agalactiae (GBS) is a major source of human perinatal diseases and bovine mastitis. Erythromycin (Ery) and tetracycline (Tet) are usually employed for preventing human and bovine infections although resistance to such agents has become common among GBS strains. Ery and Tet resistance genes are usually carried by conjugative transposons (CTns) belonging to the Tn916 family, but their presence and transferability among GBS strains have not been totally explored. Here we evaluated the presence of Tet resistance genes (tetM and tetO) and CTns among Ery-resistant (Ery-R) and Ery-susceptible (Ery-S) GBS strains isolated from human and bovine sources; and analyzed the ability for transferring resistance determinants between strains from both origins. Tet resistance and int-Tn genes were more common among Ery-R when compared to Ery-S isolates. Conjugative transfer of all resistance genes detected among the GBS strains included in this study (ermA, ermB, mef, tetM and tetO), in frequencies between 1.10-7 and 9.10-7, was possible from bovine donor strains to human recipient strain, but not the other way around. This is, to our knowledge, the first report of in vitro conjugation of Ery and Tet resistance genes among GBS strains recovered from different hosts.

Effects of erythromycin on γ-glutamyl cysteine synthetase and interleukin-1β in hyperoxia-exposed lung tissue of premature newborn rats / Efeitos da eritromicina sobre a γ-glutamil-cisteína-sintetase e a interleucina-1β no tecido pulmonar exposto à hiperóxia de ratos recém-nascidos prematuros

Objective: To explore the effect of erythromycin on hyperoxia-induced lung injury. Methods: One-day-old preterm offspring Sprague-Dawley (SD) rats were randomly divided into four groups: group 1, air + sodium chloride; group 2, air + erythromycin;group 3, hyperoxia + sodium chloride; and group 4, hyperoxia + erythromycin. At one, seven, and 14 days of exposure, glutathione (GSH) and interleukin-1 beta (IL-1 beta) were detected by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA), and bicinchoninic acid (BCA) was used to detect GSH protein. γ-glutamine-cysteine synthetase (γ-GCS) mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results: Compared with group 1, expressions of GSH and γ-GCS mRNA in group 3 were significantly increased at one and seven days of exposure (p < 0.05), but expression of γ-GCS mRNA was significantly reduced at 14 days; expression of IL-1 beta in group 3 was significantly increased at seven days of exposure (p < 0.05), and was significantly reduced at 14 days. Compared with group 3, expressions of GSH and γ-GCS mRNA in group 4 were significantly increased at one, seven, and 14 days of exposure (p < 0.05), but expressions of GSH showed a downward trend at 14 days; expression of IL-1 beta in group 4 was significantly reduced at one and seven days of exposure (p < 0.05). Conclusions: Changes in oxidant-mediated IL-1 beta and GSH are involved in the development of hyperoxia-induced lung injury. Erythromycin may up-regulate the activity of γ-GCS, increasing the expression of GSH, inhibiting the levels of oxidant-mediated IL-1 beta and alleviating hyperoxia-induced lung injury via an antioxidant effect. .

Objetivo: Explorar o efeito da eritromicina sobre lesões pulmonares induzidas por hiperóxia. Métodos: Uma prole de ratos Sprague-Dawley (SD) prematuros com um dia de vida foi dividida aleatoriamente em quatro grupos: grupo 1 ar + cloreto de sódio, grupo 2 ar + eritromicina, grupo 3 hiperóxia + cloreto de sódio e grupo 4 hiperóxia + eritromicina. Com um, sete e 14 dias de exposição, foram detectadas Glutationa (GSH) e Interleucina-1 beta (IL-1 beta) pelo ensaio imunossorvente ligado à enzima (ELISA), e o ácido bicinconinico (BCA) foi utilizado para detectar a proteína GSH. O mRNA da γ-glutamil-cisteina-sintetase (γ-GCS) foi detectado por reação em cadeia da polimerase via transcriptase reversa (RT-PCR). Resultados: Comparadas ao grupo 1, as expressões do mRNA da GSH e da γ-GCS no grupo 3 aumentaram significativamente com um e sete dias de exposição (p < 0,05), porém a expressão de mRNA da γ-GCS diminuiu significativamente aos 14 dias; a expressão de IL-1 beta no grupo 3 aumentou significativamente aos 7 dias de exposição (p < 0,05) e diminuiu significativamente aos 14 dias. Comparadas ao grupo 3, as expressões do mRNA da GSH e da γ-GCS no grupo 4 aumentaram significativamente com um, sete e 14 dias de exposição (p < 0,05), porém as expressões de GSH mostraram uma tendência de queda aos 14 dias; a expressão de IL-1 beta no grupo 4 foi reduzida significativamente com um e sete dias de exposição (p < 0,05). Conclusões: As variações de IL-1 beta e GSH mediadas por oxidantes estão envolvidas no desenvolvimento de lesão pulmonar induzida por hiperóxia. A eritromicina poderá regular positivamente a atividade da γ-GCS, aumentando a expressão de GSH, inibindo os níveis de interleucina-1beta mediada por ...

Prevalencia de portación vaginal-anal de Streptococcus agalactiae en el tercer trimestre de gestación y susceptibilidad a macrólidos y lincosamidas, en mujeres embarazadas de Clínica Alemana Temuco, Chile / Prevalence of anal-vaginal colonization of Streptococcus agalactiae in third trimester of pregnancy and susceptibility to macrolides and lincosamides, in pregnant women controlled at Clínica Alemana Temuco, Southern Chile

Introduction: Streptococcus agalactiae (GBS) is the most common agent in early neonatal sepsis. Strategies incorporating universal screening for maternal colonization show the lowest rates of perinatal infection. A significant increase in resistance to erythromycin and clindamycin by GBS has been reported around the world. There are no published data regarding prevalence and antimicrobial resistance in southern regions of Chile. Surveillance of antimicrobial resistance is essential to define the drugs of choice and alternatives, in an institution that applies prevention protocols, as Clinica Alemana Temuco (CAT) does. Objectives: to determine the prevalence of carriage of GBS in vaginal-anal areas at end of pregnancy, in CAT, Araucanía Region, Chile. To determine the susceptibility to erythromycin and clindamycin of GBS strains isolated. Results: 1,181 pregnant women were included; 167 were positive for GBS (14.4% of colonization). Sixteen were resistant to erythromycin (9.5%); 15 of these strains were also clindamycin resistant. Twenty-three of 167 were resistant to clindamycin (13.7%). Conclusions: The prevalence rate of GBS colonization was lower than previously reported in other regions of Chile. Due to the high rates of resistance to clindamycin and erythromycin it is necessary to widen the study of susceptibility to other antimicrobials to have alternatives in allergy to penicillin (primarily cefazolin and vancomycin).

Introducción: Streptococcus agalactiae es el más frecuente causal de sepsis neonatal precoz. Las estrategias con pesquisa universal de colonización materna muestran las tasas más bajas de infección perinatal. Se ha reportado un significativo aumento de resistencia de S. agalactiae a eritromicina y clindamicina en el mundo. No existen datos publicados de prevalencia y susceptibilidad antimicrobiana en las regiones del sur de Chile. La vigilancia de resistencia es fundamental para definir los antimicrobianos de elección y alternativas para prevención del cuadro en instituciones que apliquen estrategias de prevención, como Clínica Alemana Temuco (CAT). Objetivos: Determinar la prevalencia de portación de S. agalactiae en la región vaginal-anal de mujeres embarazadas de tercer trimestre en CAT. Determinar la susceptibilidad a eritromicina y clindamicina de las cepas aisladas. Resultados: 1.181 embarazadas fueron incluidas. 167 resultaron S. agalactiae (+) (14,4% de colonización). Diez y seis eran resistentes a eritromicina (9,5%). Quince de ellas también lo eran a clindamicina. Veintitrés de 167, eran resistentes a clindamicina (13,7%). Conclusiones: La tasa de prevalencia de colonización (14%) fue más baja que lo reportado anteriormente en el centro del país. Debido a la alta tasa de resistencia a clindamicina y eritromicina se hace necesario aumentar el estudio de susceptibilidad a otros antimicrobianos alternativos en pacientes alérgicas a penicilina (principalmente cefazolina y vancomicina).

Análisis molecular de la resistencia a fluoroquinolonas y macrólidos en aislados de Campylobacter jejuni de humanos, bovinos y carne de ave / Molecular analysis of fluoroquinolones and macrolides resistance in Campylobacter jejuni isolates from humans, bovine and chicken meat

Background: Campylobacter sp.- one of the leading causes of bacterial food-borne gastrointestinal illness worldwide- is increasingly resistant to fluoroquinolone and macrolide antimicrobials, which has become a major concern for public health. Objective: To describe the susceptibility patterns of Campylobacter jejuni strains to erythromycin and ciprofloxacin and to explore the origin of its resistance in human isolates. Material and Method: In this study, fifty-five ciprofloxacin and erythromycin susceptibility patterns of C. jejuni strains isolated from humans with diarrheal disease, performed by broth microdilution MIC, were compared with 55 and 44 isolates from chicken meat and bovines respectively, obtained from the Metropolitan Region, Chile. Results: Of the 55 human isolates of C. jejuni, 33 (60%) were resistant to ciprofloxacin and all were sensitive to erythromycin. Of the 55 isolates from chicken meat, 32 (58.2%) were resistant to ciprofloxacin and 1.8% were resistant to erythromycin. Of the 44 isolates of C. jejuni from cattle, 8 (18.2%) were resistant to ciprofloxacin and all were sensitive to erythromycin. Four PFGE patterns matched with certain resistance profiles and grouped isolates from human and animal. Conclusion: The findings showed continued effectiveness of erythromycin for campylobacteriosis and a high percentage of C. jejuni strains ciprofloxacin-resistant. This is interesting because it is considered that the presence of ciprofloxacin resistant strains in broiler meat can be in part the source of resistance to this antimicrobial in humans.

Introducción: El incremento de la resistencia en Campylobacter sp. (una de las principales causas de gastroenteritis bacteriana de origen alimentario) a fluoro-quinolonas y macrólidos, es un problema en salud pública. Objetivo: Conocer los patrones de susceptibilidad in vitro de Campylobacter jejuni a eritromicina y ciprofloxacina y conocer el origen de su resistencia en aislados de humanos. Material y Método: En este estudio, se compararon las susceptibilidades a ciprofloxacina y eritromicina -CIM efectuadas por microdilución en caldo- de 55 aislados de C. jejuni provenientes de humanos con enterocolitis, con 55 aislados de carne de pollo y 44 de bovinos obtenidos en la Región Metropolitana, Chile. Resultados: De 55 aislados de C. jejuni de humanos, 33(60%) presentaron resistencia a ciprofloxacina y todos presentaron susceptibilidad a eritromicina. De 55 aislados procedentes de carne de pollo, 32 (58,2%) presentaron resistencia a ciprofloxacina y un aislado resultó resistente a eritromicina (1,8%). De 44 aislados de bovinos, 8(18,2%) presentaron resistencia a ciprofloxacina y todos resultaron sensibles a eritromicina. Cuatro patrones de electroforesis a campo pulsado coincidieron en sus perfiles de resistencia y agruparon aislados de origen humano y animal. Conclusiones: Los resultados muestran que eritromicina continúa siendo efectiva para el tratamiento de la campilobacteriosis y que existe un alto porcentaje de cepas resistentes a ciprofloxacina. Se considera probable que la presencia de cepas resistentes a ciprofloxacina en la carne de pollo puede ser en parte el origen de la resistencia a este fármaco en humanos.

Increase of stress resistance in Lactococcus lactis via a novel food-grade vector expressing a shsp gene from Streptococcus thermophilus

The effects of the expression of a small heat shock protein (shsp) gene from Streptococcus thermophilus on stress resistance in Lactococcus lactis under different environmental stresses were investigated in this study. pMG36e-shsp, an expression vector, was first constructed by inserting a shsp open reading frame (ORF) cloned from S. thermophilus strain St-QC into pMG36e. Then, a food-grade expression vector, pMG-shsp, was generated by deleting the erythromycin resistance gene from pMG36e-shsp. The transformation rate of pMG-shsp was comparable to that of pMG36e-shsp when each of these two vectors was introduced into L. lactis. These results demonstrated that the shsp ORF could successfully used as a food-grade selection marker in both pMG-shsp and pMG36e-shsp. Furthermore, the growth characteristics were almost the same between L. lactis ML23 transformants harboring pMG36e or pMG-shsp. The survival rate of L. lactis ML23 expressing the shsp ORF were increased to 0.032%, 0.006%, 0.0027%, 0.03%, and 0.16% under the following environmental stresses: heat, acid, ethanol, bile salt and H2O2, respectively. These results indicated that the expression of the shsp gene in the food-grade vector pMG-shsp conferred resistance to environmental stresses without affecting the growth characteristics of L. lactis ML23.

Análisis de los fenotipos y genotipos de resistencia a eritromicina y clindamicina en cepas de Streptococcus pyogenes aisladas en Chile en un período de 10 años / Resistance phenotypes and genotypes of Streptococcus pyogenes clinical isolates in Chile over a 10-year period

Background: Macrolide and lincosamide resistance in Streptococcus pyogenes is due to the acquisition of mef, ermB and ermA genes, which confer different resistance phenotypes, namely M, MLSBconstitutive and MLSBinducible respectively. The last report of resistance in Chile was done in the period 1990-1998, in which resistance to macrolides was 5.4 percent, with M phenotype as the predominant one. Aim: To characterize the evolution of erythromycin and clindamycin resistance and their associated genes in S. pyogenes strains isolated from patients with invasive and non-invasive infections in the period 1996 to 2005. Material and Methods: Resistance to erythromycin and clindamycin was determined in 1,282 clinical isolates using the disk diffusion test. Resistant isolates were analyzed by polymerase chain reaction (PCR) for the presence of the above mentioned resistance genes. Results: Global resistance to erythromycin and clindamycin was 3.5 and 0.7 percent respectively. Eighty percent of the resistant strains possessed the M. phenotype. Conclusions: Resistance levels of S. pyogenes have decreased in Chile in the last years. Most resistant strains have M phenotype in contrast to many countries in which the MLSB constitutive phenotype is the predominant one.

Aumento de resistencia de Streptococcus agalactiae vaginal-anal en el tercer trimestre de gestación a eritromicina y clindamicina al cabo de una década de tamizaje universal / Streptococcus agalactiae increase in resistance to erythromycin and clindamycin in vaginal-anal colonization in third quarter of pregnancy in one decade of universal screening

Introduction: Streptococcus agalactiae (GBS) is the main causative agent of early perinatal sepsis. The acquisition of prevention policies has led to frequent use of intrapartum antibiotics. Surveillance of antimicrobial resistance is indispensable for defining drugs of choice and alternatives for such prophylaxis. Objectives: To determine the evolution of antimicrobial resistance of GBS from maternal colonization to drugs used in the prevention of neonatal sepsis, between 2002 and 2008. Methods: We studied 100 GBS positive vaginal and anal samples from pregnant women. Disc diffussion susceptibility method was performed for penicillin, ampicillin, cefazolin, erythromycin and clindamycin according to the Clinical and Laboratory Standards Institute (CLSI). Results: We analyzed the susceptibility of 99 strains. Seventeen were resistant to erythromycin (17.1 percent) and 13 were resistant to clindamycin (13.1 percent). Thirteen of the 17 strains resistant to erythromycin had the MLS phenotype (resistance to erythromycin and clindamycin) and 4 had the M phenotype (resistance to erythromycin only). Within the MLS phenotype, resistance was constitutive in 9 strains, and induced in 4 strains (positive D test). Compared with 2002 there was a significant increase in resistance to clindamycin (from 3.27 percent to 13.1 percent p < 0.002) and erythromycin (1.09 percent to 17 percent p < 0.001). 100 percent GSB remained sensitive to penicillin and ampicillin. Conclusions: GBS remains highly susceptible to drugs of choice for prevention of perinatal sepsis. There is a significant increase in antimicrobial resistance to clindamycin and erythromycin. Therefore, it is necessary to request susceptibility testing in GBS from third trimester of pregnancy screening in patients allergic to penicillin.

Introducción: Streptococcus agalactiae es el principal agente causal de sepsis perinatal precoz. La adquisición de políticas de prevención ha traído consigo la utilización frecuente de antimicrobianos intra-parto. La vigilancia de resistencia antimicrobiana se hace indispensable para definir el fármaco de elección y alternativas en dicha profilaxis. Nuestro centro realiza tamizaje universal desde hace 10 años. Objetivos: Determinar la evolución de la resistencia antimicrobiana de S. agalactiae de colonización materna, a los antimicrobianos utilizados en la prevención de sepsis neonatal, entre 2002 y 2008. Métodos: Se estudiaron 100 muestras vaginales-anales positivas para S. agalactiae de mujeres embarazadas, con edad gestacional de 35 a 37 semanas. Se realizó estudio de susceptibilidad in vitro por discos a penicilina, ampicilina, cefazolina, eritromicina y clindamicina, según método estandarizado por Clinical and Laboratory Standards Institute (CLSI). Resultados: Se analiza la susceptibilidad de 99 cepas. Diecisiete fueron resistentes a eritromicina (17,1 por ciento) y 13 eran resistentes a clindamicina (13,1 por ciento). De las 17 cepas resistentes a eritromicina, 13 eran fenotipo MLS y 4 del fenotipo M. Dentro del fenotipo MLS, la resistencia fue constitutiva en nueve cepas e inducible en cuatro cepas (test D positivo). En comparación con el año 2002, hubo un aumento significativo de resistencia a clindamicina (de 3,2 a 13,1 por ciento p < 0,002) y a eritromicina (de 1,09 a 17 por ciento p < 0,001). Streptococcus agalactiae se mantuvo 100 por ciento sensible a penicilina y ampicilina. Conclusiones: S. agalactiae mantiene alta sensibilidad a los antimicrobianos de elección para la prevención de sepsis neonatal y a un antimicrobiano alternativo: cefazolina. Se observó un aumento significativo de resistencia antimicrobiana a clindamicina y eritromicina. Se hace necesario, entonces, solicitar antibiograma en el tamizaje del tercer trimestre del embarazo, en pacientes alérgicas a penicilina.

Decline in Erythromycin Resistance in Group A Streptococci from Acute Pharyngitis due to Changes in the emm Genotypes Rather Than Restriction of Antibiotic Use / 대한진단검사의학회지

BACKGROUND: Group A streptococcus (GAS) is the most common cause of bacterial pharyngitis in children. Antibiotic resistance rates and emm genotypes of GAS isolated from patients with acute pharyngitis were studied in 2009. METHODS: Throat cultures were taken from 499 children with acute pharyngitis in Jinju, Korea, in 2008-2009. A total of 174 strains (34.9%) of GAS were isolated, and antimicrobial susceptibility testing was performed using the disk diffusion method. The phenotypes of macrolide resistance and macrolide resistance genes were determined. The emm genotypes were identified using PCR and sequencing. The data were compared with those acquired in 2002 in the same region. Data on the annual macrolide production were collected between 1999 and 2008. RESULTS: The resistance rates of GAS to erythromycin, clindamycin, and tetracycline were 4.6%, 2.9%, and 2.3%, respectively. The constitutive resistance rate was 62.5% for the erm(B) gene and 37.5% for the M phenotype of the mef(A) gene. emm4 was most frequently detected (28.2%), followed by emm89 (20.1%). Most of the erythromycin resistant strains had the emm28 genotype. We noted a gradual increase in macrolide production during the study period. CONCLUSIONS: The erythromycin resistance rate of GAS isolated from children with acute pharyngitis was significantly lower in 2009 (4.6%) than in 2002 (44.8%). We observed a remarkable change in the distribution of emm genotypes during the 7-yr period. The significant decline in erythromycin resistance in 2009 might be associated with a prominent decrease in the resistant genotype emm12 (3.4% in 2009 vs. 28.0% in 2002) rather than restriction of macrolide use.

Evaluation of MicroScan Synergies plus Positive Combo 3 Panels for Identification and Antimicrobial Susceptibility Testing of Staphylococcus aureus and Enterococcus Species / 대한진단검사의학회지

BACKGROUND: Few studies have evaluated the performance of the recently introduced MicroScan Synergies plus Positive Combo 3 Panels (SIPC3) (Dade Behring Inc., USA). We evaluated the clinical efficacy of the panels in identification (ID) and antimicrobial susceptibility testing (AST) of Staphylococcusaureus and enterococci. METHODS: To evaluate the panels' accuracy of identification, the results obtained using the test panels were compared with those obtained by using conventional biochemical tests in conjunction with VITEK 2 system (bio-Merieux, USA). In addition, the AST results obtained using the panels were compared with those obtained by performing CLSI broth microdilution. RESULTS: The overall agreement between the approaches for the ID of S. aureus and enterococci was 100% and 96%, respectively. The categorical and essential agreements (CA and EA) for S. aureus were 98%, each. Very major errors (VME), major errors (ME), and minor error (mE) for S. aureus were 0.45%, 0.3%, and 4.2%, respectively. The majority of VMEs were for oxacillin (8.6%), penicillin (2.0%), erythromycin (7.9%), clindamycin (3.8%), and tetracycline (4.1%). For enterococci, the CA, EA, VME, ME, and mE were 88.8%, 93.7%, 4.4%, 0%, and 2.8%, respectively. The 80.5% (29/36) of Enterococcus faecium had concordant ID with the reference. Most of the categorical errors (3 VMEs and 14 mEs) were observed for quinupristin/dalfopristin (Synercid; Catalytica Pharmaceuticals Inc., USA). CONCLUSIONS: The panels compared favorably with conventional methods for the ID and AST of S. aureus. However, we expected a better performance for ID of E. faecium and AST using Synercid.

Prevalence of mef and ermB genes in invasive pediatric erythromycin-resistant Streptococcus pneumoniae isolates from Argentina / Prevalencia de los genes mef y ermB en aislamientos invasivos de Streptococcus pneumoniae resistentes a eritromicina recuperados de pacientes pediátricos en Argentina

During the period 1993-2001, a total of 1,499 pneumococci isolates were recovered through the Argentinean surveillance of Streptococcus pneumoniae causing invasive disease in children under 6 years of age, 3.5% of which were erythromycin resistant. Among the 50 erythromycin-resistant strains available, 58% (n=29) harbored mefA/E genes (15 mefA, 30%; and 14 mefE, 28%), 34% (n=17) ermB, and 6% (n=3) both mefA/E plus ermB genes, while one isolate was negative for all the acquired genes studied. The England14-9 (42%), Poland6B-20 (20%) and Spain9v-3 (16%) clones were responsible for the emergence of pneumococcal macrolide resistance in pediatric population from Argentina.

En el marco del programa de vigilancia regional SIREVA, se analizaron 1499 aislamientos de Streptococcus pneumoniae causantes de enfermedad invasiva en menores de 6 años, recuperados entre 1993 y 2001. Se detectó un 3,5% de resistencia a eritromicina. De los 50 aislamientos resistentes a eritromicina que pudieron ser estudiados, el 58% (n=29) tenían los genes mefA/E (15 mefA, 30% y 14 mefE, 28%), el 34% (n=17) el gen ermB y el 6% (n=3) la combinación de genes mefA/E y ermB. Sólo un aislamiento fue negativo para todos los genes analizados. Los clones internacionales England14-9, Poland6B-20 y Spain9v-3 representaron el 78% del total de aislamientos resistentes (42, 20 y 16%, respectivamente) y se consideraron los responsables de la emergencia de la resistencia a macrólidos entre los neumococos que afectan a la población pediátrica de Argentina.

Inducible clindamycin resistance among clinical isolates of Staphylococci.

INTRODUCTION: Clinical failure of clindamycin therapy has been reported due to multiple mechanisms that confer resistance to macrolide, lincosamide and streptogramin antibiotics. This study was undertaken to detect the presence of inducible clindamycin resistance among clinical isolates of staphylococci. MATERIALS AND METHODS: The detection of inducible clindamycin resistance was performed by D-test using erythromycin and clindamycin discs as per CDC guidelines. RESULTS: Among the 244 clinical isolates of staphylococci studied, 32 (13.1%) showed inducible clindamycin resistance and belonged to the MLSBi phenotype. Among the MLS B i phenotypes, 10 isolates were methicillin-resistant Staphylococcus aureus (38.4% of the total MRSA), 16 were methicillin-sensitive Staphylococcus aureus (12.9% of the total MSSA) and 6 were coagulase-negative staphylococci (6.3% of the total CONS). CONCLUSION: The test for inducible resistance to clindamycin should be included in the routine antibiotic susceptibility testing, as it will help in guiding therapy.