Biofilms

Surfaces are important microbial habitats for at least two reasons. For one, nutrients
adsorb to surfaces and thus often contain more resources than are available to planktonic
cells (cells that live a floating existence). And second , surfaces are areas to which
microbial cells can attach. Attachment is a means for cells to remain in a favorable
habitat and not be washed away. As a consequence, microbial numbers and activities are
typically quite high on surface.
A surface may also be a nutrient, such as a particle of organic matter. On such
surfaces the attached microorganisms catabolize nutrient directly from the surface of the
particle. For example, plant roots become heavily colonized by soil bacteria living on
organic exudates from the plant, and fluorescent stains can be used to detect and
enumerate them.

Biofilms
As bacterial cells grow on surfaces they tend to form biofilms- assemblages of bacterial
cells attached to a surface and enclosed in an adhesive matrix excreted by the cells. The
study begin when Van Leeuwenhoek, using his simple microscopes, first observed
microorganisms on tooth surfaces and can be credited with the discovery of microbial
biofilms. Heukelekian and Heller observed the bottle effect for marine
microorganisms, i.e., bacterial growth and activity were substantially enhanced by the
incorporation of a surface to which these organisms could attach. Zobell observed that
the number of bacteria on surfaces was dramatically higher than in the surrounding
medium (in this case, seawater). However, a detailed examination of biofilms would
await the electron microscope, which allowed high-resolution photomicroscopy at much
higher magnifications than did the light microscope. Jones et al. used scanning and
transmission electron microscopy to examine biofilms on trickling filters in a
wastewater treatment plant and showed them to be composed of a variety of organisms
(based on cell morphology). By using a specific polysaccharide-stain called Ruthenium
red and coupling this with osmium tetroxide fixative, these researchers were also able to
show that the matrix material surrounding and enclosing cells in these biofilms was
polysaccharide.
As early as 1973, Characklis studied microbial slimes in industrial water systems
and showed that they were not only very tenacious but also highly resistant to
disinfectants such as chlorine. Based on observations of dental plaque and sessile
communities in mountain streams, Costerton et al. in 1978 put forth a theory of biofilms
that explained the mechanisms whereby microorganisms adhere to living and nonliving

materials and the benefits accrued by this ecologic niche. Since that time, the studies of
biofilms in industrial and ecologic settings and in environments more relevant for public
health have basically paralleled each other. Much of the work in the last 2 decades has
relied on tools such as scanning electron microscopy (SEM) or standard microbiologic
culture techniques for biofilm characterization. Two major thrusts in the last decade
have dramatically impacted our understanding of biofilms: the utilization of the confocal
laser scanning microscope to characterize biofilm ultrastructure, and an investigation of
the genes involved in cell adhesion and biofilm formation.
The matrix of biofilms is typically a mixture of polysacharides, but can contain
proteins and even nucleic acid. Biofilms trap nutrients for microbial growth and help
prevent the detachment of cells on surfaces present in flowing systems (Figure 1).

Figure 1. Example of microbial biofilms

Biofilms typically contain several porous layers, and the cells in each layer can
be examined by scanning laser confocal microscopy. Biofilms may contain only one or
two species or, more commonly, several species of bacteria. The biofilm that forms
around a tooth surface, for example, contain a several hundred different phylotypes,
including species of both Bacteria and Archae.

Biofilm Formation
Formation of a biofilm begins with the attachment of free-floating microorganisms to a
surface. These first colonists adhere to the surface initially through weak, reversible
adhesion via van der Waals forces. If the colonists are not immediately separated from
the surface, they can anchor themselves more permanently using cell adhesion structures
such as pili. Hydrophobicity also plays an important role in determining the ability of

bacteria to form biofilms, as those with increased hydrophobicity have reduced

repulsion between the extracellular matrix and the bacterium.
Some species are not able to attach to a surface on their own but are sometimes
able to anchor themselves to the matrix or directly to earlier colonists. It is during this
colonization that the cells are able to communicate via quorum sensing using products
such as acylated homoserine. Some bacteria are unable to form biofilms as successfully
due to their limited motility. Nonmotile bacteria cannot recognize the surface or
aggregate together as easily as motile bacteria. Once colonization has begun, the biofilm
grows through a combination of cell division and recruitment. Polysaccharide matrices
typically enclose bacterial biofilms. In addition to the polysaccharides, these matrices
may also contain material from the surrounding environment, including but not limited
to minerals, soil particles, and blood components, such as erythrocytes and fibrin. The
final stage of biofilm formation is known as dispersion, and is the stage in which the
biofilm is established and may only change in shape and size. The development of a
biofilm may allow for an aggregate cell colony (or colonies) to be increasingly antibiotic
resistant. Cell-cell communication or quorum sensing has been shown to be involved in
the formation of biofilm in several bacterial species (Figure 2).

Figure 2. Biofilm formation

Biofilms: Advantages and Control

The biofilm lifestyle confers many advantages on microbial populations, and once
formed, biofilms can be very difficult to control.

Why Do Bacteria Form Biofilms?

At least four reason underlie the formation of biofilms.
1. Biofilms are means of microbial self-defense. Biofilm resist physical forces that
could sweep away unattached cells. Moreover, biofilms resist phagocytosis by
cells of the immune system and the penetration of toxic molecules, such as
antibiotics. All of these advantages improve the chances for survival of cells in
the biofilm.
2. Biofilms formation allows cells to remain in a favorable niche. Biofilms attached
to nutrient-rich surfaces, such as animal tissues or to surfaces in flowing systems,
such as a rock in a stream.
3. Biofilms form because they alow bacterial cells to live in close association with
each other.
4. Biofilms seem to be the typical way bacterial cells grow in nature.
Biofilm Control
Biofilms have significant implications in human medicine and commerce. In the body,
bacterial cells within a biofilm are protected from attack by the immune system, and
antibiotics and other antimicrobial agents often fail to pierce the biofilm. Biofilms have
been implicated in medical and dental condition, including periodontal disease, kidney
stone, tuberculosis, Legionnaires disease, and Staphylococcus infections.
Biofilm control is big business and thus far, only a limited number of tools exist
to fight biofilms. Collectively, industries commit huge financial resources to treating
pipes and other surfaces to keep them free of biofilms. New antimicrobial agents that
can be penetrate biofilms, as well as drugs that prevent biofilm formation by interfering
with intercellular communication, are being developed. A class of chemicals called
furanones, for example, has shown promise as biofilm preventatives in tests on abiotic
surfaces. Furanones are also stable and some are relatively nontoxic, so they may have
applications as antibiofilm agents in human medicine as well.

References
Donlan, Rodney M. 2002. Biofilms: Microbial Life on Surfaces. EID Journals. 8: 881890
Maerdigan, et al. 2009. Brock Biology of Microorganisms. USA: Pearson Education Inc.