Characterization of multi-drug tolerant persister cells in Streptococcus suis

BMC Microbiology

Peter Valentin-Weigand²  Ralph Goethe²  Ralph Bertram¹  Daniela Willms²  Jörg Willenborg² 

[1] Department of Microbial Genetics, University of Tübingen, Tübingen, Germany;Institute of Microbiology, University of Veterinary Medicine, Hannover, Germany

关键词: Antibiotics;    Multidrug tolerance;    Persister cells;    Streptococcus suis;   

Others  :  1141090
DOI  :  10.1186/1471-2180-14-120

 received in 2014-02-25, accepted in 2014-05-06,  发布年份 2014

【 摘 要 】

Background

Persister cells constitute a subpopulation of dormant cells within a microbial population which are genetically identical but phenotypically different to regular cells. Notably, persister cells show an elevated tolerance to antimicrobial agents. Thus, they are considered to represent a microbial ‘bet-hedging’ strategy and are of particular importance in pathogenic bacteria.

Results

We studied the ability of the zoonotic pathogen Streptococcus (S.) suis to form multi-drug tolerant variants and identified persister cells dependent on the initial bacterial growth phase. We observed lower numbers of persisters in exponential phase cultures than in stationary growth phase populations. S. suis persister cells showed a high tolerance to a variety of antibiotics, and the phenotype was not inherited as tested with four passages of S. suis populations. Furthermore, we provide evidence that the persister phenotype is related to expression of genes involved in general metabolic pathways since we found higher numbers of persister cells in a mutant strain defective in the catabolic arginine deiminase system as compared to its parental wild type strain. Finally, we observed persister cell formation also in other S. suis strains and pathogenic streptococcal species.

Conclusions

Taken together, this is the first study that reports multi-drug tolerant persister cells in the zoonotic pathogen S. suis.

【 授权许可】

   
2014 Willenborg et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Bigger J: Treatment of staphylococcal infections with penicillin by intermittent sterilisation. Lancet 1944, 244(6320):497-500.
• [2]Balaban NQ, Gerdes K, Lewis K, McKinney JD: A problem of persistence: still more questions than answers? Nat Rev Microbiol 2013, 11:587-591.
• [3]Wiuff C, Zappala RM, Regoes RR, Garner KN, Baquero F, Levin BR: Phenotypic tolerance: antibiotic enrichment of noninherited resistance in bacterial populations. Antimicrob Agents Chemother 2005, 49:1483-1494.
• [4]Lewis K: Persister cells. Annu Rev Microbiol 2010, 64:357-372.
• [5]Kussell E, Leibler S: Phenotypic diversity, population growth, and information in fluctuating environments. Science 2005, 309:2075-2078.
• [6]Balaban NQ, Merrin J, Chait R, Kowalik L, Leibler S: Bacterial persistence as a phenotypic switch. Science 2004, 305:1622-1625.
• [7]Dhar N, McKinney JD: Microbial phenotypic heterogeneity and antibiotic tolerance. Curr Opin Microbiol 2007, 10:30-38.
• [8]Johnson PJ, Levin BR: Pharmacodynamics, population dynamics, and the evolution of persistence in Staphylococcus aureus. PLoS Genet 2013, 9:e1003123.
• [9]Fauvart M, De Groote VN, Michiels J: Role of persister cells in chronic infections: clinical relevance and perspectives on anti-persister therapies. J Med Microbiol 2011, 60:699-709.
• [10]Moyed HS, Bertrand KP: hipA, a newly recognized gene of Escherichia coli K-12 that affects frequency of persistence after inhibition of murein synthesis. J Bacteriol 1983, 155:768-775.
• [11]Gerdes K, Maisonneuve E: Bacterial persistence and toxin-antitoxin loci. Annu Rev Microbiol 2012, 66:103-123.
• [12]Amato SM, Orman MA, Brynildsen MP: Metabolic control of persister formation in Escherichia coli. Mol Cell 2013, 50:475-487.
• [13]Nguyen D, Joshi-Datar A, Lepine F, Bauerle E, Olakanmi O, Beer K, McKay G, Siehnel R, Schafhauser J, Wang Y, Britigan BE, Singh PK: Active starvation responses mediate antibiotic tolerance in biofilms and nutrient-limited bacteria. Science 2011, 334:982-986.
• [14]Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K: Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 2004, 230:13-18.
• [15]Lechner S, Lewis K, Bertram R: Staphylococcus aureus persisters tolerant to bactericidal antibiotics. J Mol Microbiol Biotechnol 2012, 22:235-244.
• [16]Brooun A, Liu S, Lewis K: A dose–response study of antibiotic resistance in Pseudomonas aeruginosa biofilms. Antimicrob Agents Chemother 2000, 44:640-646.
• [17]Keren I, Minami S, Rubin E, Lewis K: Characterization and transcriptome analysis of Mycobacterium tuberculosis persisters. MBio 2011, 2:e00100-e00111.
• [18]Wakamoto Y, Dhar N, Chait R, Schneider K, Signorino-Gelo F, Leibler S, McKinney JD: Dynamic persistence of antibiotic-stressed mycobacteria. Science 2013, 339:91-95.
• [19]Shapiro JA, Nguyen VL, Chamberlain NR: Evidence for persisters in Staphylococcus epidermidis RP62a planktonic cultures and biofilms. J Med Microbiol 2011, 60:950-960.
• [20]Singh R, Ray P, Das A, Sharma M: Role of persisters and small-colony variants in antibiotic resistance of planktonic and biofilm-associated Staphylococcus aureus: an in vitro study. J Med Microbiol 2009, 58:1067-1073.
• [21]Cohen NR, Lobritz MA, Collins JJ: Microbial persistence and the road to drug resistance. Cell Host Microbe 2013, 13:632-642.
• [22]Spoering AL, Lewis K: Biofilms and planktonic cells of Pseudomonas aeruginosa have similar resistance to killing by antimicrobials. J Bacteriol 2001, 183:6746-6751.
• [23]Lewis K: Persister cells, dormancy and infectious disease. Nat Rev Microbiol 2007, 5:48-56.
• [24]Keren I, Shah D, Spoering A, Kaldalu N, Lewis K: Specialized persister cells and the mechanism of multidrug tolerance in Escherichia coli. J Bacteriol 2004, 186:8172-8180.
• [25]Lewis K: Multidrug tolerance of biofilms and persister cells. Curr Top Microbiol Immunol 2008, 322:107-131.
• [26]Leung V, Levesque CM: A stress-inducible quorum-sensing peptide mediates the formation of persister cells with noninherited multidrug tolerance. J Bacteriol 2012, 194:2265-2274.
• [27]Arends JP, Zanen HC: Meningitis Caused by Streptococcus suis in Humans. Rev Infect Dis 1988, 10:131-137.
• [28]Chanter N, Jones PW, Alexander TJ: Meningitis in pigs caused by Streptococcus suis - a speculative review. Vet Microbiol 1993, 36:39-55.
• [29]Clifton-Hadley FA, Alexander TJ: The carrier site and carrier rate of Streptococcus suis type II in pigs. Vet Rec 1980, 107:40-41.
• [30]Gottschalk M, Xu J, Calzas C, Segura M: Streptococcus suis: a new emerging or an old neglected zoonotic pathogen? Future Microbiol 2010, 5:371-391.
• [31]Mai NT, Hoa NT, Nga TV, Linh LD, Chau TT, Sinh DX, Phu NH, Chuong LV, Diep TS, Campbell J, Nghia HD, Minh TN, Chau NV, de Jong MD, Chinh NT, Hien TT, Farrar J, Schultsz C: Streptococcus suis meningitis in adults in Vietnam. Clin Infect Dis 2008, 46:659-667.
• [32]Wertheim HF, Nguyen HN, Taylor W, Lien TT, Ngo HT, Nguyen TQ, Nguyen BN, Nguyen HH, Nguyen HM, Nguyen CT, Dao TT, Nguyen TV, Fox A, Farrar J, Schultsz C, Nguyen HD, Nguyen KV, Horby P: Streptococcus suis, an important cause of adult bacterial meningitis in northern Vietnam. PLoS One 2009, 4:e5973.
• [33]Baums CG, Verkuhlen GJ, Rehm T, Silva LM, Beyerbach M, Pohlmeyer K, Valentin-Weigand P: Prevalence of Streptococcus suis genotypes in wild boars of Northwestern Germany. Appl Environ Microbiol 2007, 73:711-717.
• [34]Sanchez DR V, Fernandez-Garayzabal JF, Briones V, Iriso A, Dominguez L, Gottschalk M, Vela AI: Genetic analysis of Streptococcus suis isolates from wild rabbits. Vet Microbiol 2013, 165:483-486.
• [35]Varela NP, Gadbois P, Thibault C, Gottschalk M, Dick P, Wilson J: Antimicrobial resistance and prudent drug use for Streptococcus suis. Anim Health Res Rev 2013, 14:68-77.
• [36]Tan JH, Yeh BI, Seet CS: Deafness due to haemorrhagic labyrinthitis and a review of relapses in Streptococcus suis meningitis. Singapore Med J 2010, 51:e30-e33.
• [37]Fulde M, Willenborg J, De Greeff A, Benga L, Smith HE, Valentin-Weigand P, Goethe R: ArgR is an essential local transcriptional regulator of the arcABC operon in Streptococcus suis and is crucial for biological fitness in an acidic environment. Microbiology 2011, 157:572-582.
• [38]Gruening P, Fulde M, Valentin-Weigand P, Goethe R: Structure, regulation, and putative function of the arginine deiminase system of Streptococcus suis. J Bacteriol 2006, 188:361-369.
• [39]Willenborg J, Fulde M, De Greeff A, Rohde M, Smith HE, Valentin-Weigand P, Goethe R: Role of glucose and CcpA in capsule expression and virulence of Streptococcus suis. Microbiology 2011, 157:1823-1833.
• [40]Chen C, Tang J, Dong W, Wang C, Feng Y, Wang J, Zheng F, Pan X, Liu D, Li M, Song Y, Zhu X, Sun H, Feng T, Guo Z, Ju A, Ge J, Dong Y, Sun W, Jiang Y, Wang J, Yan J, Yang H, Wang X, Gao GF, Yang R, Wang J, Yu J: A glimpse of streptococcal toxic shock syndrome from comparative genomics of S. suis 2 Chinese isolates. PLoS One 2007, 2:e315.
• [41]Allgaier A, Goethe R, Wisselink HJ, Smith HE, Valentin-Weigand P: Relatedness of Streptococcus suis isolates of various serotypes and clinical backgrounds as evaluated by macrorestriction analysis and expression of potential virulence traits. J Clin Microbiol 2001, 39:445-453.
• [42]Betriu C, Gomez M, Sanchez A, Cruceyra A, Romero J, Picazo JJ: Antibiotic resistance and penicillin tolerance in clinical isolates of group B streptococci. Antimicrob Agents Chemother 1994, 38:2183-2186.
• [43]Pichichero ME, Casey JR: Systematic review of factors contributing to penicillin treatment failure in Streptococcus pyogenes pharyngitis. Otolaryngol Head Neck Surg 2007, 137:851-857.
• [44]Entenza JM, Caldelari I, Glauser MP, Francioli P, Moreillon P: Importance of genotypic and phenotypic tolerance in the treatment of experimental endocarditis due to Streptococcus gordonii. J Infect Dis 1997, 175:70-76.
• [45]Orman MA, Brynildsen MP: Establishment of a method to rapidly assay bacterial persister metabolism. Antimicrob Agents Chemother 2013, 57:4398-4409.
• [46]Luidalepp H, Joers A, Kaldalu N, Tenson T: Age of inoculum strongly influences persister frequency and can mask effects of mutations implicated in altered persistence. J Bacteriol 2011, 193:3598-3605.
• [47]Bizzini A, Entenza JM, Moreillon P: Loss of penicillin tolerance by inactivating the carbon catabolite repression determinant CcpA in Streptococcus gordonii. J Antimicrob Chemother 2007, 59:607-615.
• [48]Bradely JJ, Mayhall CG, Dalton HP: Incidence and characteristics of antibiotic-tolerant strains of Staphylococcus aureus. Antimicrob Agents Chemother 1978, 13:1052-1057.
• [49]Sader HS, Flamm RK, Farrell DJ, Jones RN: Daptomycin activity against uncommonly isolated streptococcal and other gram-positive species groups. Antimicrob Agents Chemother 2013, 57:6378-6380.
• [50]Francois B, Gissot V, Ploy MC, Vignon P: Recurrent septic shock due to Streptococcus suis. J Clin Microbiol 1998, 36:2395.
• [51]Bonifait L, Grignon L, Grenier D: Fibrinogen induces biofilm formation by Streptococcus suis and enhances its antibiotic resistance. Appl Environ Microbiol 2008, 74:4969-4972.
• [52]Olson ME, Ceri H, Morck DW, Buret AG, Read RR: Biofilm bacteria: formation and comparative susceptibility to antibiotics. Can J Vet Res 2002, 66:86-92.
• [53]Brisebois LM, Charlebois R, Higgins R, Nadeau M: Prevalence of Streptococcus suis in four to eight week old clinically healthy piglets. Can J Vet Res 1990, 54:174-177.
• [54]MacInnes JI, Gottschalk M, Lone AG, Metcalf DS, Ojha S, Rosendal T, Watson SB, Friendship RM: Prevalence of Actinobacillus pleuropneumoniae, Actinobacillus suis, Haemophilus parasuis, Pasteurella multocida, and Streptococcus suis in representative Ontario swine herds. Can J Vet Res 2008, 72:242-248.
• [55]Amass SF, Wu CC, Clark LK: Evaluation of antibiotics for the elimination of the tonsillar carrier state of Streptococcus suis in pigs. J Vet Diagn Invest 1996, 8:64-67.
• [56]Smith HE, Veenbergen V, Van der Velde J, Damman M, Wisselink HJ, Smits MA: The cps genes of Streptococcus suis serotypes 1, 2, and 9: development of rapid serotype-specific PCR assays. J Clin Microbiol 1999, 37:3146-3152.
• [57]Schubert A, Zakikhany K, Schreiner M, Frank R, Spellerberg B, Eikmanns BJ, Reinscheid DJ: A fibrinogen receptor from group B Streptococcus interacts with fibrinogen by repetitive units with novel ligand binding sites. Mol Microbiol 2002, 46:557-569.
• [58]Rohde M, Muller E, Chhatwal GS, Talay SR: Host cell caveolae act as an entry-port for group A streptococci. Cell Microbiol 2003, 5:323-342.
• [59]Molinari G, Talay SR, Valentin-Weigand P, Rohde M, Chhatwal GS: The fibronectin-binding protein of Streptococcus pyogenes, SfbI, is involved in the internalization of group A streptococci by epithelial cells. Infect Immun 1997, 65:1357-1363.