Chapter 18 : The Influence of Biofilms in the Biology of Plasmids

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The natural state for many bacteria is not growth in liquid culture, but, rather, living as a community attached to a surface. These bacterial communities, termed biofilms, exist in the natural world as well as in the human host. The Centers for Disease Control and Prevention and the National Institutes of Health have estimated that approximately 65 to 80% of human infections are biofilm related. A recent burgeoning area of research has examined the role of plasmids in biofilms, including the effect of conjugative plasmid transfer on biofilm formation, as well as the role of biofilms in plasmid dissemination. In addition, heterogeneity in the biofilm population in terms of plasmid carriage has also been demonstrated. Most published studies of plasmid biology and conjugation in biofilms have focused on Gram-negative spp. such as and . In this article, we will review these studies in relation to recent work focusing on effects of biofilm growth on plasmid-related functions such as gene transfer and antimicrobial resistance in Gram-positive pathogens such as and .

Citation: Cook L, Dunny G. 2015. The Influence of Biofilms in the Biology of Plasmids, p 315-323. In Tolmasky M, Alonso J (ed), Plasmids: Biology and Impact in Biotechnology and Discovery. ASM Press, Washington, DC. doi: 10.1128/microbiolspec.PLAS-0012-2013
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Figure 1

Formation of a bacterial biofilm. Bacterial biofilm development involves three stages. Initial attachment of single group or small groups of bacteria to a surface, often aided by attachment structures such as pili. Growth of these attached cells as well as attachment of additional cells increases the biomass of the biofilm. Concurrently, the bacteria produce an extracellular matrix made of up various components including DNA, protein, and polysaccharides that help the biofilm retain its structure and keep the biofilm cells attached to the surface and to each other. During and after the formation of a large biofilm, individual cells or even large pieces of the biofilm may break off. These detached cells may go on to live a planktonic lifestyle or seed new biofilms. Dark lines indicate the components of the matrix used to attach cells to each other and the surface such as eDNA ( ), while orange extracellular material indicates other matrix components used to retain biofilm structure and surface attachment.

Citation: Cook L, Dunny G. 2015. The Influence of Biofilms in the Biology of Plasmids, p 315-323. In Tolmasky M, Alonso J (ed), Plasmids: Biology and Impact in Biotechnology and Discovery. ASM Press, Washington, DC. doi: 10.1128/microbiolspec.PLAS-0012-2013
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Figure 2

The interplay between conjugation and biofilm development. (Top) A model proposed by Ghigo ( ), based on his analysis of conjugation between strains. Planktonic populations of donor cells (green), carrying plasmids such as R1, whose conjugation functions are normally repressed, contain a few spontaneously depressed individuals. When these depressed cells encounter a biofilm containing recipient cells (white), they can attach via their sex pili (red) and transfer the plasmid. In newly generated transconjugants, there is a transient period where repression of conjugation is not operative. This can be followed by “epidemic spread” of the plasmid through the biofilm population, and the associated production of sex pili also can increase the biofilm biomass directly. (Bottom) In , expression of conjugation is regulated by peptide-mating pheromones produced by recipient cells. In the scenario depicted on the left, the pheromone produced by recipient cells (white) in a biofilm turns on expression of conjugation in planktonic donor cells (blue) in close proximity, and the resulting synthesis of pheromone-induced surface adhesins (thick, gray layer) promotes both an increase in biofilm resulting from increased attachment of planktonic cells, and also leads to plasmid transfer within the biofilm. In the right, the development of a mixed biofilm as a result of attachment of both donors and recipients to the same surface may allow for signaling and conjugation between sessile donor and recipient cells in close proximity ( ).

Citation: Cook L, Dunny G. 2015. The Influence of Biofilms in the Biology of Plasmids, p 315-323. In Tolmasky M, Alonso J (ed), Plasmids: Biology and Impact in Biotechnology and Discovery. ASM Press, Washington, DC. doi: 10.1128/microbiolspec.PLAS-0012-2013
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Figure 3

Three mechanistic models that could account for increased ratios of plasmids/chromosome in a fraction of biofilm cells. Figures depict actively growing “planktonic-like” diplococcal cells prior to cell division. Each cell half contains two copies of the chromosome (represented by thick lines), where a new round of replication has initiated on one chromosome (indicated by the “bubble”). Each cell half also contains three copies of a plasmid, with the dashed lines indicating rolling circle replication initiated by either a single-stranded replication initiator protein or a conjugative relaxase. (It should be noted that some plasmids showing increased copy number in biofilms actually use a “theta” mode of replication similar to the chromosome [ ]). Each diagram contains one diplococcal cell (left) with a plasmid copy number similar to that of planktonic cells and a second cell with an altered copy number, representing a subpopulation of the biofilm community. The three diagrams illustrate three possible mechanisms by which the plasmid/chromosome ratio could be increased in these subpopulations. (A) Secretion of eDNA into the biofilm matrix by a fraction of cells reduces the ratio of chromosome/plasmid. (B) Biofilm growth reduces chromosomal copy number in subpopulations by inhibiting initiation of chromosome replication. (C) In a subpopulation of biofilm cells, changes in cellular physiology could disrupt normal mechanisms limiting conjugative nicking or vegetative plasmid replication initiation, leading to increases in copy number. Additionally, for some well-studied plasmids, such as ColE1, blockage of chromosome replication initiation (B) can lead to “runaway” plasmid replication ( ). Based on published results ( ), all of these models are postulated to operate in the early stages of biofilm development, when the adherent cells are still growing, and there is no significant death or lysis.

Citation: Cook L, Dunny G. 2015. The Influence of Biofilms in the Biology of Plasmids, p 315-323. In Tolmasky M, Alonso J (ed), Plasmids: Biology and Impact in Biotechnology and Discovery. ASM Press, Washington, DC. doi: 10.1128/microbiolspec.PLAS-0012-2013
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1. Barnes AM,, Ballering KS,, Leibman RS,, Wells CL,, Dunny GM . 2012. Enterococcus faecalis produces abundant extracellular structures containing DNA in the absence of cell lysis during early biofilm formation. MBio 3 : e00193-12. doi:10.1128/mBio.00193-12 [PubMed] [CrossRef]
2. Branda SS,, Chu F,, Kearns DB,, Losick R,, Kolter R . 2006. A major protein component of the Bacillus subtilis biofilm matrix. Mol Microbiol 59 : 1229 1238.[PubMed] [CrossRef]
3. Costerton JW,, Cheng KJ,, Geesey GG,, Ladd TI,, Nickel JC,, Dasgupta M,, Marrie TJ . 1987. Bacterial biofilms in nature and disease. Annu Rev Microbiol 41 : 435 464.[PubMed] [CrossRef]
4. Flemming H-C,, Wingender J . 2010. The biofilm matrix. Nat Rev Microbiol 8 : 623 633.[PubMed]
5. Reisner A,, Haagensen JA,, Schembri MA,, Zechner EL,, Molin S . 2003. Development and maturation of Escherichia coli K-12 biofilms. Mol Microbiol 48 : 933 946.[PubMed] [CrossRef]
6. Pratt LA,, Kolter R . Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol Microbiol 30 : 285 293.[PubMed] [CrossRef]
7. Sauer K,, Camper AK,, Ehrlich GD,, Costerton JW,, Davies DG . 2002. Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J Bacteriol 184 : 1140 1154.[PubMed] [CrossRef]
8. Singh PK,, Schaefer AL,, Parsek MR,, Moninger TO,, Welsh MJ,, Greenberg EP . 2000. Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Nature 407 : 762 764.[PubMed] [CrossRef]
9. Chai Y,, Chu F,, Kolter R,, Losick R . 2008. Bistability and biofilm formation in Bacillus subtilis . Mol Microbiol 67 : 254 263.[PubMed] [CrossRef]
10. Aspiras MB,, Ellen RP,, Cvitkovitch DG . 2004. ComX activity of Streptococcus mutans growing in biofilms. FEMS Microbiol Lett 238 : 167 174.[PubMed]
11. Li T,, Lau P,, Lee J,, Ellen R,, Cvitkovitch D . 2001. Natural genetic transformation of Streptococcus mutans growing in biofilms. J Bacteriol 183 : 897 908.[PubMed] [CrossRef]
12. Li YH,, Tang N,, Aspiras MB,, Lau PC,, Lee JH,, Ellen RP,, Cvitkovitch DG . 2002. A quorum-sensing signaling system essential for genetic competence in Streptococcus mutans is involved in biofilm formation. J Bacteriol 184 : 2699 2708.[CrossRef]
13. Eaton RE,, Jacques NA . 2010. Deletion of competence-induced genes over-expressed in biofilms caused transformation deficiencies in Streptococcus mutans . Mol Oral Microbiol 25 : 406 417.[PubMed] [CrossRef]
14. Burmolle M,, Bahl MI,, Jensen LB,, Sorensen SJ,, Hansen LH . 2008. Type 3 fimbriae, encoded by the conjugative plasmid pOLA52, enhance biofilm formation and transfer frequencies in Enterobacteriaceae strains. Microbiology 154 : 187 195.[PubMed] [CrossRef]
15. Ghigo J . 2001. Natural conjugative plasmids induce bacterial biofilm development. Nature 412 : 442 445.[PubMed] [CrossRef]
16. D’Alvise PW,, Sjoholm OR,, Yankelevich T,, Jin Y,, Weurtz S,, Smets BF . 2010. TOL plasmid carriage enhances biofilm formation and increases extracellular DNA content in Pseudomonas putida KT2440. FEMS Microbiol Lett 312 : 84 92.[PubMed] [CrossRef]
17. Ong CL,, Beatson SA,, McEwan AG,, Schembri MA . 2009. Conjugative plasmid transfer and adhesion dynamics in an Escherichia coli biofilm. Appl Environ Microbiol 75 : 6783 6791.[PubMed] [CrossRef]
18. Molin S,, Tolker-Nielsen T . 2003. Gene transfer occurs with enhanced efficiency in biofilms and induced enhanced stabilisation of the biofilm structure. Curr Opin Biotechnol 14 : 255 261.[PubMed] [CrossRef]
19. Yang X,, Ma Q,, Wood TK . 2008. The R1 conjugative plasmid increases Escherichia coli biofilm formation through an envelope stress response. Appl Environ Microbiol 74 : 2690 2699.[PubMed] [CrossRef]
20. Reisner A,, Holler BM,, Molin S,, Zechner EL . 2006. Synergistic effects in mixed Escherichia coli biofilms: conjugative plasmid transfer drives biofilm expansion. J Bacteriol 188 : 3582 3588.[PubMed] [CrossRef]
21. May T,, Okabe S . 2008. Escherichia coli harboring a natural IncF conjugative F plasmid develops complex mature biofilms by stimulating synthesis of colanic acid and Curli. J Bacteriol 190 : 7479 7490.[PubMed] [CrossRef]
22. Castonguay MH,, van der Schaaf S,, Koester W,, Krooneman J,, van der Meer W,, Harmsen H,, Landini P . 2006. Biofilm formation by Escherichia coli is stimulated by synergistic interactions and co-adhesion mechanisms with adherence-proficient bacteria. Res Microbiol 157 : 471 478.[PubMed] [CrossRef]
23. Bahl MI,, Hansen LH,, Goesmann A,, Sorensen SJ . 2007. The multiple antibiotic resistance IncP-1 plasmid pKJK5 isolated from a soil environment is phylogenetically divergent from members of the previously established alpha, beta and delta sub-groups. Plasmid 58 : 31 43.[PubMed] [CrossRef]
24. Roder HL,, Hansen LH,, Sorensen SJ,, Burmolle M . 2013. The impact of the conjugative IncP-1 plasmid pKJK5 on multispecies biofilm formation is dependent on the plasmid host. FEMS Microbiol Lett 344 : 186 192.[PubMed] [CrossRef]
25. Hausner M,, Wuertz S . 1999. High rates of conjugation in bacterial biofilms as determined by quantitative in situ analysis. Appl Environ Microbiol 65 : 3710 3713.[PubMed]
26. Sorensen SJ,, Bailey M,, Hansen LH,, Kroer N,, Wuertz S . 2005. Studying plasmid horizontal transfer in situ: a critical review. Nat Rev Microbiol 3 : 700 710.[PubMed] [CrossRef]
27. Savage VJ,, Chopra I,, O’Neill AJ . 2013. Staphylococcus aureus biofilms promote horizontal transfer of antibiotic resistance. Antimicrob Agents Chemother 57 : 1968 1970.[PubMed] [CrossRef]
28. Keen NT,, Tamaki S,, Kobayashi D,, Trollinger D . 1988. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene 70 : 191 197.[PubMed] [CrossRef]
29. Licht TR,, Christensen BB,, Krogfelt KA,, Molin S . 1999. Plasmid transfer in the animal intestine and other dynamic bacterial populations: the role of community structure and environment. Microbiology 145( Pt 9) : 2615 2622.[PubMed]
30. Ehlers LJ,, Bouwer EJ . 1999. Rp4 plasmid transfer among species of Pseudomonas in a biofilm reactor. Water Sci Tech 39 : 163 171.[CrossRef]
31. Krol JE,, Wojtowicz AJ,, Rogers LM,, Heuer H,, Smalla K,, Krone SM,, Top EM . 2013. Invasion of E. coli biofilms by antibiotic resistance plasmids. Plasmid 70 : 110 119.[PubMed] [CrossRef]
32. Davies D,, Geesey G . 1995. Regulation of the alginate biosynthesis gene algC in Pseudomonas aeruginosa during biofilm development in continuous culture. Appl Environ Microbiol 61 : 860 867.[PubMed]
33. May T,, Ito A,, Okabe S . 2009. Induction of multidrug resistance mechanism in Escherichia coli biofilms by interplay between tetracycline and ampicillin resistance genes. Antimicrob Agents Chemother 53 : 4628 4639.[PubMed] [CrossRef]
34. Cook L,, Chatterjee A,, Barnes A,, Yarwood J,, Hu W-S,, Dunny G . 2011. Biofilm growth alters regulation of conjugation by a bacterial pheromone. Mol Microbiol 81 : 1499 1510.[PubMed] [CrossRef]
35. Cook LC,, Dunny GM . 2013. Effects of biofilm growth on plasmid copy number and expression of antibiotic resistance genes in Enterococcus faecalis . Antimicrob Agents Chemother 57 : 1850 1856.[PubMed] [CrossRef]
36. Clewell DB . 1972. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol 110 : 667 676.[PubMed]
37. Togna PA,, Shuler ML,, Wilson DB . 1993. Effects of plasmid copy number and runaway plasmid replication on overproduction and excretion of B-lactamase from Escherichia coli . Biotechnol Prog 9 : 31 39.[PubMed] [CrossRef]
38. Uhlin BE,, Nordström K . 1978. A runaway-replication mutant of plasmid R1drd-19: temperature-dependent loss of copy number control. Mol Gen Genet 165 : 167 179.[PubMed] [CrossRef]
39. Ma H,, Katzenmeyer KN,, Bryers JD . 2013. Non-invasive in situ monitoring and quantification of TOL plasmid segregational loss within Pseudomonas putida biofilms. Biotechnol Bioeng 110 : 2949 2958.[PubMed] [CrossRef]
40. Imran M,, Jones D,, Smith H . 2005. Biofilms and the plasmid maintenance question. Math Biosci 193 : 183 204.[PubMed] [CrossRef]
41. Teodosio JS,, Simoes M,, Mergulhao FJ . 2012. The influence of nonconjugative Escherichia coli plasmids on biofilm formation and resistance. J Appl Microbiol 113 : 373 382.[PubMed] [CrossRef]
42. Baur B,, Hanselmann K,, Schlimme W,, Jenni B . 1996. Genetic transformation in freshwater: Escherichia coli is able to develop natural competence. Appl Environ Microbiol 62 : 3673 3678.[PubMed]
43. Tsen SD,, Fang SS,, Chen MJ,, Chien JY,, Lee CC,, Tsen DH . 2002. Natural plasmid transformation in Escherichia coli . J Biomed Sci 9 : 246 252.[PubMed]
44. Maeda S,, Sawamura A,, Matsuda A . 2004. Transformation of colonial Escherichia coli on solid media. FEMS Microbiol Lett 236 : 61 64.[PubMed] [CrossRef]
45. Maeda S,, Ito M,, Ando T,, Ishimoto Y,, Fujisawa Y,, Takahashi H,, Matsuda A,, Sawamura A,, Kato S . 2006. Horizontal transfer of nonconjugative plasmids in a colony biofilm of Escherichia coli . FEMS Microbiol Lett 255 : 115 120.[PubMed] [CrossRef]
46. Berge MJ,, Kamgoue A,, Martin B,, Polard P,, Campo N,, Claverys JP . 2013. Midcell recruitment of the DNA uptake and virulence nuclease, EndA, for pneumococcal transformation. PLoS Pathog 9 : e1003596. doi:10.1371/journal.ppat.1003596. [PubMed] [CrossRef]

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