Commensal Streptococcus agalactiaeisolated from patients seen at University Hospital of Londrina, Paraná, Brazil: capsular types, genotyping, antimicrobial susceptibility and virulence determinants

Eliane Saori Otaguiri1, Ana Elisa Belotto Morguette1, Eliandro Reis Tavares1, Pollyanna Myrella Capela dos Santos1, Alexandre Tadachi Morey1, Juscélio Donizete Cardoso1, Márcia Regina Eches Perugini2, Lucy Megumi Yamauchi1, Sueli Fumie Yamada‐Ogatta1
1Laboratório de Biologia Molecular de Microrganismos, Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Brazil
2Laboratório de Microbiologia Clínica, Departamento de Patologia, Análises Clínicas e Toxicológicas, Centro de Ciências da Saúde, Universidade Estadual de Londrina, Londrina, Brazil

Tóm tắt

Abstract Background Streptococcus agalactiae or Group B Streptococci (GBS) have the ability to access various host sites, which reflects its adaptability to different environments during the course of infection. This adaptation is due to the expression of virulence factors that are involved with survival, invasion and bacterial persistence in the host. This study aimed to characterize GBS isolates from women of reproductive age seen at University Hospital of Londrina, according to capsular typing, genetic relatedness, antimicrobial susceptibility profile and occurrence of virulence determinants. Results A total of 83 GBS isolates were enrolled in this study. Capsular types Ia (42.2%), II (10.8%), III (14.5%) and V (30.1%) were identified in most GBS. One isolate each was classified as type IX and non-typeable. A total of 15 multiple locus variable number of tandem repeat analysis (MLVA) types were identified among the isolates, seven were singletons and eight were represented by more than four isolates. All isolates were susceptible to penicillin, ampicillin, cefepime, cefotaxime, chloramphenicol, levofloxacin and vancomycin. Resistance to erythromycin and clindamycin was observed in 19.3 and 13.3% of isolates, respectively. All isolates resistant to clindamycin were simultaneously resistant to erythromycin and were distributed in the capsular types III and V. One isolate showed the constitutive macrolide-lincosamide-streptogramin B (cMLSB) phenotype and ten showed the inducible MLSB (iMLSB) phenotype. The mechanism of resistance to erythromycin and clindamycin more prevalent among these isolates was mediated by the gene ermA, alone or in combination with the gene ermB. The isolates displaying resistance only to erythromycin belonged to capsular type Ia, and showed the M phenotype, which was mediated by the mefA/E gene. All isolates harbored the gene hylB and at least one pilus variant, PI-1, PI-2a or PI-2b. Although cylE was observed in all GBS, four isolates were classified as gamma-hemolytic and carotenoid pigment non-producers. Conclusions Our results indicate the potential virulence of commensal GBS isolates, reinforcing the need for continued screening for this bacterium to prevent infections. The distribution of capsular and pili antigens, and MLVA profiles was also identified, which may contribute to the development of new strategies for the prevention and treatment of GBS infection.

Từ khóa


Tài liệu tham khảo

McCord N, Owen P, Powls A, Lunan B: A complete audit cycle of intrapartum group B streptococcus prophylaxis. Health Bull (Edinb). 2001, 59: 263-267.

Krohn MA, Hillier SL, Baker CJ: Maternal peripartum complications associated with vaginal group B streptococci colonization. J Infect Dis. 1999, 179: 1410-1415. 10.1086/314756.

Phares CR, Lynfield R, Farley MM, Mohle-Boetani J, Harrison LH, Petit S, Craig AS, Schaffner W, Zansky SM, Gershman K, et al: Epidemiology of invasive group B streptococcal disease in the United States, 1999–2005. JAMA. 2008, 299: 2056-2065. 10.1001/jama.299.17.2056.

Schuchat A: Group B streptococcal disease in newborns: A global perspective on prevention. Biomed Pharmacother. 1995, 49: 19-25. 10.1016/0753-3322(96)82573-X.

Verani JR, Schrag SJ: Group B streptococcal disease in infants: Progress in prevention and continued challenges. Clin Perinatol. 2010, 37: 375-392. 10.1016/j.clp.2010.02.002.

Verani JR, McGee L, Schrag SJ: Prevention of perinatal group B streptococcal disease-revised guidelines from CDC, 2010. MMWR Recomm Rep. 2010, 59: 1-36.

Edmond KM, Kortsalioudaki C, Scott S, Schrag SJ, Zaidi AK, Cousens S, Heath PT: Group B streptococcal disease in infants aged younger than 3 months: Systematic review and meta-analysis. Lancet. 2012, 379: 547-556. 10.1016/S0140-6736(11)61651-6.

Edwards MS, Baker CJ: Group B streptococcal infections in elderly adults. Clin Infect Dis. 2005, 41: 839-847. 10.1086/432804.

Skoff TH, Farley MM, Petit S, Craig AS, Schaffner W, Gershman K, Harrison LH, Lynfield R, Mohle-Boetani J, Zansky S, et al: Increasing burden of invasive group B streptococcal disease in nonpregnant adults, 1990–2007. Clin Infect Dis. 2009, 49: 85-92. 10.1086/599369.

Duarte RS, Bellei BC, Miranda OP, Brito MA, Teixeira LM: Distribution of antimicrobial resistance and virulence-related genes among Brazilian group B streptococci recovered from bovine and human sources. Antimicrob Agents Chemother. 2005, 49: 97-103. 10.1128/AAC.49.1.97-103.2005.

Palmeiro JK, Dalla-Costa LM, Fracalanzza SE, Botelho AC, da Silva Nogueira K, Scheffer MC, de Almeida Torres RS, de Carvalho NS, Cogo LL, Madeira HM: Phenotypic and genotypic characterization of group B streptococcal isolates in southern Brazil. J Clin Microbiol. 2010, 48: 4397-4403. 10.1128/JCM.00419-10.

Correa AB, Silva LG, Pinto Tde C, Oliveira IC, Fernandes FG, Costa NS, Mattos MC, Fracalanzza SE, Benchetrit LC: The genetic diversity and phenotypic characterisation of Streptococcus agalactiae isolates from Rio de Janeiro, Brazil. Mem Inst Oswaldo Cruz. 2011, 106: 1002-1006. 10.1590/S0074-02762011000800017.

Nakamura PA, Schuab RBB, Neves FP, Pereira CF, Paula GR, Barros RR: Antimicrobial resistance profiles and genetic characterisation of macrolide resistant isolates of Streptococcus agalactiae. Mem Inst Oswaldo Cruz. 2011, 106: 119-122. 10.1590/S0074-02762011000200001.

Nagano N, Nagano Y, Toyama M, Kimura K, Tamura T, Shibayama K, Arakawa Y: Nosocomial spread of multidrug-resistant group B streptococci with reduced penicillin susceptibility belonging to clonal complex 1. J Antimicrob Chemother. 2012, 67: 849-856. 10.1093/jac/dkr546.

Capanna F, Emonet SP, Cherkaoui A, Irion OP, Schrenzel J, MartinezdeTejada B: Antibiotic resistance patterns among group B Streptococcus isolates: Implications for antibiotic prophylaxis for early-onset neonatal sepsis. Swiss Med Wkly. 2013, 143: 0-

Leclercq R: Mechanisms of resistance to macrolides and lincosamides: Nature of the resistance elements and their clinical implications. Clin Infect Dis. 2002, 34: 482-492. 10.1086/324626.

Clancy J, Petitpas J, Dib-Hajj F, Yuan W, Cronan M, Kamath AV, Bergeron J, Retsema JA: Molecular cloning and functional analysis of a novel macrolide-resistance determinant, mefA, from Streptococcus pyogenes. Mol Microbiol. 1996, 22: 867-879. 10.1046/j.1365-2958.1996.01521.x.

Cieslewicz MJ, Chaffin D, Glusman G, Kasper D, Madan A, Rodrigues S, Fahey J, Wessels MR, Rubens CE: Structural and genetic diversity of group B streptococcus capsular polysaccharides. Infect Immun. 2005, 73: 3096-3103. 10.1128/IAI.73.5.3096-3103.2005.

Slotved HC, Kong F, Lambertsen L, Sauer S, Gilbert GL: Serotype IX, a Proposed New Streptococcus agalactiae Serotype. J Clin Microbiol. 2007, 45: 2929-2936. 10.1128/JCM.00117-07.

Murayama SY, Seki C, Sakata H, Sunaoshi K, Nakayama E, Iwata S, Sunakawa K, Ubukata K: Capsular type and antibiotic resistance in Streptococcus agalactiae isolates from patients, ranging from newborns to the elderly, with invasive infections. Antimicrob Agents Chemother. 2009, 53: 2650-2653. 10.1128/AAC.01716-08.

Madzivhandila M, Adrian PV, Cutland CL, Kuwanda L, Madhi SA: Distribution of pilus islands of group B streptococcus associated with maternal colonization and invasive disease in South Africa. J Med Microbiol. 2013, 62: 249-253. 10.1099/jmm.0.052951-0.

Marques MB, Kasper DL, Pangburn MK, Wessels MR: Prevention of C3 deposition by capsular polysaccharide is a virulence mechanism of type III group B streptococci. Infect Immun. 1992, 60: 3986-3993.

Lauer P, Rinaudo CD, Soriani M, Margarit I, Maione D, Rosini R, Taddei AR, Mora M, Rappuoli R, Grandi G, Telford JL: Genome analysis reveals pili in Group B Streptococcus. Science. 2005, 309: 105-10.1126/science.1111563.

Sharma P, Lata H, Arya DK, Kashyap AK, Kumar H, Dua M, Ali A, Johri AK: Role of pilus proteins in adherence and invasion of Streptococcus agalactiae to the lung and cervical epithelial cells. J Biol Chem. 2013, 288: 4023-4034. 10.1074/jbc.M112.425728.

Rinaudo CD, Rosini R, Galeotti CL, Berti F, Necchi F, Reguzzi V, Ghezzo C, Telford JL, Grandi G, Maione D: Specific involvement of pilus type 2a in biofilm formation in group B Streptococcus. PLoS One. 2010, 5: e9216-10.1371/journal.pone.0009216.

Maisey HC, Quach D, Hensler ME, Liu GY, Gallo RL, Nizet V, Doran KS: A group B streptococcal pilus protein promotes phagocyte resistance and systemic virulence. FASEB J. 2008, 22: 1715-1724. 10.1096/fj.07-093963.

Pritzlaff CA, Chang JC, Kuo SP, Tamura GS, Rubens CE, Nizet V: Genetic basis for the beta-haemolytic/cytolytic activity of group B Streptococcus. Mol Microbiol. 2001, 39: 236-247. 10.1046/j.1365-2958.2001.02211.x.

Doran KS, Chang JC, Benoit VM, Eckmann L, Nizet V: Group B streptococcal beta-hemolysin/cytolysin promotes invasion of human lung epithelial cells and the release of interleukin-8. J Infect Dis. 2002, 185: 196-203. 10.1086/338475.

Liu GY, Doran KS, Lawrence T, Turkson N, Puliti M, Tissi L, Nizet V: Sword and shield: linked group B streptococcal beta-hemolysin/cytolysin and carotenoid pigment function to subvert host phagocyte defense. Proc Natl Acad Sci U S A. 2004, 101: 14491-14496. 10.1073/pnas.0406143101.

Baker JR, Pritchard DG: Action pattern and substrate specificity of the hyaluronan lyase from group B streptococci. Biochem J. 2000, 348 (Pt 2): 465-471.

Benchetrit LC, Fracalanzza SE, Peregrino H, Camelo AA, Sanches LA: Carriage of Streptococcus agalactiae in women and neonates and distribution of serological types: a study in Brazil. J Clin Microbiol. 1982, 15: 787-790.

Haguenoer E, Baty G, Pourcel C, Lartigue MF, Domelier AS, Rosenau A, Quentin R, Mereghetti L, Lanotte P: A multi locus variable number of tandem repeat analysis (MLVA) scheme for Streptococcus agalactiae genotyping. BMC Microbiol. 2011, 11: 171-10.1186/1471-2180-11-171.

Radtke A, Lindstedt BA, Afset JE, Bergh K: Rapid multiple-locus variant-repeat assay (MLVA) for genotyping of Streptococcus agalactiae. J Clin Microbiol. 2010, 48: 2502-2508. 10.1128/JCM.00234-10.

Uh Y, Kim HY, Jang IH, Hwang GY, Yoon KJ: Correlation of serotypes and genotypes of macrolide-resistant Streptococcus agalactiae. Yonsei Med J. 2005, 46: 480-483. 10.3349/ymj.2005.46.4.480.

Rosini R, Rinaudo CD, Soriani M, Lauer P, Mora M, Maione D, Taddei A, Santi I, Ghezzo C, Brettoni C, et al: Identification of novel genomic islands coding for antigenic pilus-like structures in Streptococcus agalactiae. Mol Microbiol. 2006, 61: 126-141. 10.1111/j.1365-2958.2006.05225.x.

Martins ER, Andreu A, Melo-Cristino J, Ramirez M: Distribution of Pilus islands in streptococcus agalactiae that cause human infections: Insights into evolution and implication for vaccine development. Clin Vaccine Immunol. 2013, 20: 313-316. 10.1128/CVI.00529-12.

Forquin MP, Tazi A, Rosa-Fraile M, Poyart C, Trieu-Cuot P, Dramsi S: The putative glycosyltransferase-encoding gene cylJ and the group B Streptococcus (GBS)-specific gene cylK modulate hemolysin production and virulence of GBS. Infect Immun. 2007, 75: 2063-2066. 10.1128/IAI.01565-06.

Merritt K, Jacobs NJ: Characterization and incidence of pigment production by human clinical group B streptococci. J Clin Microbiol. 1978, 8: 105-107.

Milligan TW, Baker CJ, Straus DC, Mattingly SJ: Association of elevated levels of extracellular neuraminidase with clinical isolates of type III group B streptococci. Infect Immun. 1978, 21: 738-746.

Granlund M, Oberg L, Sellin M, Norgren M: Identification of a novel insertion element, IS1548, in group B streptococci, predominantly in strains causing endocarditis. J Infect Dis. 1998, 177: 967-976. 10.1086/515233.

Horan TC, Andrus M, Dudeck MA: CDC/NHSN surveillance definition of health care-associated infection and criteria for specific types of infections in the acute care setting. Am J Infect Control. 2008, 36: 309-332. 10.1016/j.ajic.2008.03.002.

de Paris F, Machado AB, Gheno TC, Ascoli BM, Oliveira KR, Barth AL: Group B Streptococcus detection: comparison of PCR assay and culture as a screening method for pregnant women. Braz J Infect Dis. 2011, 15: 323-327.

Imperi M, Pataracchia M, Alfarone G, Baldassarri L, Orefici G, Creti R: A multiplex PCR assay for the direct identification of the capsular type (Ia to IX) of Streptococcus agalactiae. J Microbiol Methods. 2010, 80: 212-214. 10.1016/j.mimet.2009.11.010.

Hunter PR, Gaston MA: Numerical index of the discriminatory variability of typing systems: An application of Simpson’s index of diversity. J Clin Microbiol. 1988, 26: 2465-2466.

CLSI: Performance standards for antimicrobial susceptibility testing. Twenty-second informational supplement (M100-S22). 2012, Wayne, PA: Clinical and Laboratory Standards Institute

Seppala H, Nissinen A, Yu Q, Huovinen P: Three different phenotypes of erythromycin-resistant Streptococcus pyogenes in Finland. J Antimicrob Chemother. 1993, 32: 885-891. 10.1093/jac/32.6.885.