A biofilm is any group of microorganisms in which cells stick to each other and

often these cells adhere to a surface. These adherent cells are frequently embed
ded within a self-produced matrix of extracellular polymeric substance (EPS). Bi
ofilm extracellular polymeric substance, which is also referred to as slime (alt
hough not everything described as slimeis a biofilm), is a polymeric conglomerat
ion generally composed of extracellular DNA, proteins, and polysaccharides. Biof
ilms may form on living or non-living surfaces and can be prevalent in natural,
industrial and hospital settings.[2][3] The microbial cells growing in a biofilm
are physiologically distinct from planktonic cells of the same organism, which,
by contrast, are single-cells that may float or swim in a liquid medium.
Microbes form a biofilm in response to many factors, which may include cellular
recognition of specific or non-specific attachment sites on a surface, nutrition
al cues, or in some cases, by exposure of planktonic cells to sub-inhibitory con
centrations of antibiotics.[4][5] When a cell switches to the biofilm mode of gr
owth, it undergoes a phenotypic shift in behavior in which large suites of genes
are differentially regulated.[6]
An iridescent biofilm on the surface of a fish tank.
Formation of a biofilm begins with the attachment of free-floating microorganism
s to a surface. While still not fully understood, it is thought that the first c
olonists of a biofilm adhere to the surface initially through weak, reversible a
dhesion via van der Waals forces and hydrophobic effects.[7][8] 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 pl
ays an important role in determining the ability of bacteria to form biofilms, a
s those with increased hydrophobicity have reduced repulsion between the extrace
llular matrix and the bacterium.[9]
Some species are not able to attach to a surface on their own but are instead ab
le to anchor themselves to the matrix or directly to earlier colonists. It is du
ring this colonization that the cells are able to communicate via quorum sensing
(QS) using products such as N-acyl homoserine lactone (AHL). Some bacteria are
unable to form biofilms as successfully due to their limited motility. Non-motil
e bacteria cannot recognize the surface or aggregate together as easily as motil
e bacteria.[9] Once colonization has begun, the biofilm grows through a combinat
ion 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.
[9] The final stage of biofilm formation is known as dispersion, and is the stag
e 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 resistant to antibiotics. Cell-cell communication or quorum
sensing has been shown to be involved in the formation of biofilm in several ba
cterial species.[10]
Five stages of biofilm development: (1) Initial attachment, (2) Irreversible att
achment, (3) Maturation I, (4) Maturation II, and (5) Dispersion. Each stage of
development in the diagram is paired with a photomicrograph of a developing P. a
eruginosa biofilm. All photomicrographs are shown to the same scale.
There are five stages of biofilm development (see illustration at right):
Initial attachment.
Irreversible attachment.
Maturation I.
Maturation II.

Some biofi lms have been found to contain water channels that help distribute nutrients and signalling molecules.[13][14] Recent evidence has sh own that a fatty acid messenger.In conclusion. but highly sensitive towards iron stress. e. The social structure (cooperation/competition) within a biofilm depends highly on the different species present. Secreted by Pseudomonas aerug inosa. wit h its particular toxicity effects on bacterial viability. Nitric oxide has the potential for the treatment of patients th at suffer from chronic infections caused by biofilms. Enzy mes that degrade the biofilm extracellular matrix. as compared with planktonic cells.[18] It is generally assumed that cells dispersed from biofilms immediately go into t he planktonic growth phase. Biofilms can contain many different types of microorganism . A large proportion of the EPS is more or less strongly hydrated. emergence of bacterial resistance to QS inhibitors and its adverse effects must be evalua ted in-depth. The EPS matrix is an important key to the evolutio nary success of biofilms. it appears that targeting QS to mitigate membrane biofouling is a promising technology for the enhancement of efficiency and performance in membr ane systems. particularly in high humidity climates.[15] Nitric oxide has also been shown to tr igger the dispersal of biofilms of several bacteria species[16][17] at sub-toxic concentrations. each group performs speciali zed metabolic functions. Given suffi cient resources for growth.g. Rev. However. this compound induces cyclo heteromorphic cells in several species of bac teria and the yeast Candida albicans. may play a role in biofilm dispersal. One reason is that it traps extracellular enzymes and keeps them in close proximity to the cells. However. although they can form as floating mats on liquid surfaces and al so on the surface of leaves. 623-633). biofilms can become fossilized (Stromatolites). one example is cel lulose which is produced by a range of microorganisms.[19][20] Hence. protozoa. the matrix represents an exter nal digestion system and allows for stable synergistic microconsortia of differe nt species (Wingender and Flemming. recent studies have shown that the physiolo gy of dispersed cells from Pseudomonas aeruginosa biofilms is highly different f rom those of planktonic and biofilm cells. Furthermore.[19] Properties[edit] Biofilms are usually found on solid substrates submerged in or exposed to an aqu eous solution.[citation needed] Dispersal[edit] Dispersal of cells from the biofilm colony is an essential stage of the biofilm life cycle. however. In fact. current validation methods for QS must be improved and optimized. fungi andalgae. However. the dispersal process is a unique stage during the transition from biofilm to planktonic lifestyle in bacteria. Dispersed cells are found to be highly virulent against macrophages an d Caenorhabditis elegans.[11][12] Biofilm matrix degrad ing enzymes may be useful as anti-biofilm agents. some organisms will form single-species films under certain conditions. Future advances in membrane biofouling mitigation based on QS can be expected from further fundamental research. such as dispersin Band deoxyr ibonuclease. Microbiol. Nat. EPS is an abbreviation for either extracellular polymeric sub stance or exopolysaccharide.[22] This matrix is strong enough that under certain condi tions. the EPS matrix consists not only of polysaccharid es but also of proteins (which may be the major component in environmental and w aste water biofilms) and nucleic acids.[21] Extracellular matrix[edit] The biofilm is held together and protected by a matrix of secreted polymeric com pounds called EPS. Dispersal enables biofilms to spread and colonize new surfaces. in order to mitigate membrane biofouling . This matrix encases the c ells within it and facilitates communication among them through biochemical sign als as well as gene exchange. 8. although the latter one only refers to the polysacc haride moiety of EPS. Different strategies have b een demonstrated to target QS activity. Thus. hydrophobic EPS also occur. Results from recent investigations verified the existence of a corr elation between QS activity and membrane biofouling. cis-2-decenoic acid. is capable of inducing dis persion and inhibiting growth of biofilm colonies. Bacteria living in a biofilm usually have significantly different properties fro . bacteria. a biofilm will quickly grow to be macroscopic (visib le to the naked eye). archaea.

which extract and digest organic com pounds. bacteria are mainly responsible for removal of organic matter (BOD). In fact. Thermophilic bacteria in the outflow of Mickey Hot Springs. Biofilms c an form inside water and sewagepipes and cause clogging and corrosion. microbially influenced corrosion). This resistance to antibiotics in both stationary-pha se cells and biofilms may be due to the presence ofpersister cells.[26] Habitat[edit] Mats of bacterial biofilm color the hot springs inYellowstone National Park. for example.e. a s the dense extracellular matrix and the outer layer of cells protect the interi or of the community.Oregon. Biofilms on floors and counters can make sanitation difficult in food preparation areas.or heating-water systems are known to reduce heat t ransfer. prolonging voyages and consuming fuel. have mechanisms by which they can adhere to surfaces and to each other. • Bacterial adhesion to boat hulls serves as the foundation for biofouling of seagoing vessels. • Biofilms can grow in the most extreme environments: from. • Biofilms can also be harnessed for constructive purposes. it is easier for other mari ne organisms such as barnacles to attach. briny waters of hot springs ranging from very acidic to very a lkaline. • In the human environment.m free-floating bacteria of the same species. not only bacteri a and archaea. For instance.. Biofilms will form on virtually every non-shedding surface in a non-steri le aqueous (or very humid) environment. t he extremely hot. Slow sand . • Biofilms in cooling.[25]Enzymatic degradation of ext racellular DNA can weaken the biofilm structure and release microbial cells from the surface.[28] can lead to substantial corrosion problems.[23] Lateral gene transfer is greatly facilitated in biofilms and leads to a more stable biofilm structure. as the dense and protected environ ment of the film allows them to cooperate and interact in various ways. However. the biofilm form of Pseudomonas aeruginosa has no greater resistance to antimicr obials than do stationary-phase planktonic cells. Time in dry dock for refitting and repainting reduces the productivity of shipping assets. and the u seful life of ships is also reduced due to corrosion and mechanical removal (scr aping) of marine organisms from ships' hulls. Biofilms are ubiquitous. bi ofilms are important components offood chains in rivers and streams and are graz ed by the aquatic invertebrates upon which many fish feed. although when the biofilm is c ompared to logarithmic-phase planktonic cells. including pathogens and other microorganisms. Such fouling can reduce maximum vessel speed by up to 20%. while protozoa and rotifers are mainly responsible for removal of suspended solids (SS). Corrosio n is mainly due to abiotic factors. the biofilm does have greater res istance to antimicrobials. at least 20% of corrosion is caused by microorganisms that are attached to the metal subsurface (i. however. Nearly every species of microorganism. In such biofilms.[24] Extracellular DNA is a major structural component of many different microbial biofilms. to frozen glaciers.[27] • Biofilms in marine engineering systems. biofilms can grow in showers very easily since they provide a moist and warm environment for the biofilm to thrive. biofilms are not always less susceptible to antibiotics. Once a film of bacteria forms. For example. m any sewage treatment plants include a secondary treatment stage in which waste w ater passes over biofilms grown on filters. such as pipelines of the offshor e oil and gas industry. In some cases antibiotic resistance can be increased a thou sandfold. One bene fit of this environment is increased resistance to detergents and antibiotics. The longest raised mat area is about half a meter long. approximately 20 mm thick. • Biofilms can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water.

Biofilms and infectious diseases[edit] Biofilms have been found to be involved in a wide variety of microbial infection s in the body. wher e they may cause tooth decay and gum disease. formation of dental plaque. What we regard as clean wa ter is effectively a waste material to these microcellular organisms. middle-ear infections. includi ng Lactobacillus plantarum and Lactococcus lactis) and gram-negative species (e. and are still forming today. unlike the control . especially of cyanobacteria. and in the soil. e. This is especially important because the appendix holds a mass amount of these bacterial bio-films. catheter infections. by one estimate 80% of all infections. Stromatolites include some of the most anci ent records of life on Earth. including complex organic waste and renew able biomass.[40][41] More recently it has been noted that bacterial biofilm s may impair cutaneous wound healing and reduce topical antibacterial efficiency in healing or treating infected skin wounds. Pierce's Disease of grapes. g. Staphylococcus spp.[34] This discovery helps to distinguish the possible functi on of the appendix and the idea that the appendix can help reinoculate the gut w ith good gut flora. Pseudomonas putida. Bacillus sp p. • Biofilms can help eliminate petroleum oil from contaminated oceans or ma rine systems. spring or river sources for drinking purposes.[31] • Biofilms are used in microbial fuel cells (MFCs) to generate electricity from a variety of starting materials.g. including gram-positive (e. and the majority of their n atural isolates form biofilms. and infections of permanent indwelling devices such as joint prostheses and heart valves. Although many techniq ues have developed to identify planktonic bacteria in viable wounds.[43] It has recently been shown that biofilms are present on the removed tissue of 80 % of patients undergoing surgery for chronic sinusitis. Future studies ar e needed to find means of identifying and monitoring biofilm colonization at the bedside to permit timely initiation of treatment.[38] gingivitis.[39] and les s common but more lethal processes such as endocarditis.[35] Biofilms are formed by bacteria that colonize plants. P seudomonas fluorescens. • Biofilms are found on the surface of and inside plants. This was supported mainly with the fact t hat the two most abundantly produced molecules by the immune system also support bio-film production and are associated with the bio-films developed in the gut. the so-called hydrocarbonoclastic bacteria (HCB). Escherichia coli. coating contact lenses. The oil is eliminated by the hydrocarbon-degrading activities of m icrobial communities. in particular by a remarkable recently discovered group of specialists.[36] For other species in disease-associated biofilms see below. or Pseudomonas aeruginosa). and related pseudomonads which are common plant-associat ed bacteria found on leaves. infections in cystic fi brosis. The patients with biofil ms were shown to have been denuded of cilia and goblet cells. and Bacterial Spot of plants such as peppers and tomatoes.[37] Infectious processes i n which biofilms have been implicated include common problems such as bacterial vaginosis.[30] Examples of crop diseases related t o biofilms include Citrus Canker.[29] • Stromatolites are layered accretionary structures formed in shallow wate r by the trapping.[36] Several nitrogen-fixing symbionts of legumes such as Rhizobium leguminosarum and Sinorhizobium meliloti form biofilms on legu me roots and other inert surfaces.[42] Early detection of biofilms in wounds is crucial to successful chronic wound management. Listeria monocytogenes. roots. few have be en able to quickly and accurately identify bacterial biofilms. They can either contribute to crop disease or. urinary tract infections.filters rely on biofilm development in the same way to filter surface water from lake. and lactic acid bacteria.[3][32][33] • Recent studies in 2003 discovered that the immune system supports bio-fi lm development in the large intestine. as in the case of nitrogen-fixing Rhizobium on ro ots. • Biofilms are present on the teeth of most animals as dental plaque. Taxonomic diversity[edit] Many different bacteria form biofilms.g. exist symbiotically with the plant. binding and cementation of sedimentary grains by microbial bi ofilms.

tooth rushing).[52] Dietary car ohydrates can cause a dramatic decrease in pH in oral iofilms to values of 4 and elow (acid stress). the sign and symptom eing a carious lesion.to 600-fold higher than S. and cariogenic micro iological population develops and is maint ained y frequent consumption of fermenta le dietary car ohydrate.e. from tissues with allergy-inflammations. or d uring the normal course of anti iotic therapy. em edded in salivary polymers and micro ial extracellular products. the cu ltures were negative though the bacteria were present.[45] Biofilms can also be formed on the inert surfaces of implanted devices such as c atheters. This su -therapeutic level of anti iotic may result from the use of anti iotics as growth promoters in agriculture. e. sugar intake) and frequent removal of the iofilm (i. from an evolutionary point of view. mutans can survive the frequent acid s . mutans cells are genetica lly transformed at a rate 10. Competence can lead to genetic transformation. containing S. the competence regulon is induced. leading to resistance to eing killed y acid. The iofilm formation induced y low-level methicillin was inhi ited y DNase.g.[56][57] Gen etic competence is the a ility of a cell to take up DNA released y another cell . suggesting that the su -therapeuti c levels of anti iotic also induce extracellular DNA release.[52] A pH of 4 at ody temperature of 37 °C causes depurination of DNA. The speci es of bacteria from interoperative cultures did not correspond to the bacteria s pecies in the biofilm on the respective patient's tissue.[49] Dental plaque[edit] Dental plaque is an oral iofilm that adheres to the teeth and consists of many species of oth acteria and fungi (such as Streptococcus mutans and Candida al icans). mutans and related oral streptococci. The accumulation of microorganisms su jects the teeth and gingival tissues to high c oncentrations of acterial meta olites which results in dental disease.[50] The iofilm on the surface of teeth is frequently su ject to oxidative stress[51 ] and acid stress. [46] New staining techniques are being developed to differentiate bacterial cells gro wing in living animals.[47] Research has shown that sub-therapeutic levels of β-lactam anti iotics induce iof ilm formation in Staphylococcus aureus. leaving apurinic (AP) sites in DNA. mutans cells r eside in an actively growing iofilm.[52] As pointed out y Michod et al. The resulting activity shift in the iofilm (and resulting acid production within the iofilm . A peptide pheromone quorum sensing signaling system in S. at the tooth surface) is associated with an im alance etween demineralization and remineralisation leading to net mineral loss within dental hard tissues (en amel and then dentin). dental caries can e prevented and arrested.[44] Biofilms were also found on samples from two of 10 healthy controls mentioned.[54] The Dental plaque iofilm can result in the disease dental caries if it is allow ed to develop over time. An ecologic shift away from alanced populations within the dental iofilm is driven y certain (cariogenic) micro iological population s eginning to dominate when the environment favours them. f avored under conditions of high cell density and/or stress where there is maxima l opportunity for interaction etween the competent cell and the DNA released fr om near y donor cells. the creation of the tragedy of the commons in pa thogenic micro es may provide advanced therapeutic ways for chronic infections c aused y iofilms via genetically engineered invasive cheaters who can invade wi ld-types ‘cooperators’ of pathogenic acteria until cooperator populations go to ext inction or overall population ‘cooperators and cheaters ’ go to extinction. This system is optimally expressed when S.[56] When the iofilm.e. mutans growing as freefloating planktonic cells suspended in liquid. [53] especially loss of guanine. By preventin g the Dental plaque iofilm from maturing or y returning it ack to a non-cario genic state.[58] It appears that S. is su ject ed to acid stress. transformation in act erial pathogens likely provides for effective and efficient recom inational repa ir of DNA damages. The shift to an Acido genic. In other words..[48] Moreover. aciduric. Biofilm grown S. prosthetic cardiac valves and intrauterine devices.[55] This can e achiev ed though the ehavioural step of reducing the supply of fermenta le car ohydrat es (i. mutans includes the Co mpetence Stimulating Peptide (CSP) that controls genetic competence. a form of sexual interaction.s without biofilms who had normal cilia and goblet cell morphology.

microorganisms attach to the surfaces and iofilms develop internall y. pneumoniae is the main cause of community-acquired pneumonia and meningitis i n children and the elderly. ut also increases virulence in pneumoni a and meningitis. pneumoniae in a iofilm have the survival advantage that they can more easily take up transforming DNA from near y cells in the iofilm to us e for recom inational repair of oxidative damages in their DNA. and soil.tress in oral iofilms. Salmonella is als o found in the seafood industry where iofilms form from seafood orne pathogens .[61] 60-70% of nosocomial or hospital acquired infections are associated with the imp lantation of a iomedical device.[61] Biofilms in the food industry[edit] Biofilms have ecome pro lematic in several food industries due to the a ility t o form on plants and during industrial processes. It has een proposed that competence development and iofilm formation is an ada ptation of S. and of septicemia in HIV-infected persons. pne umoniae can also secrete an enzyme (murein hydrolase) that destroys non-competen t cells (fratricide) causing DNA to e released into the surrounding medium for potential use y the competent cells. through the recom inational repair provided y competence and transformation.[61] This leads to 2 million cases annually in the U. No matter the sophistication.S. it ecomes even harder to treat. Competent S.. animal manure. genes are specifically expressed that respond to oxi dative stress and induce competence. CSP also functions as a quorum-sensing peptide.[64] Along with economic pro lems iofilm formation on food pos es a health risk to consumers due to the a ility to make the food more resistant to disinfectants [62] As a result. yet this industry c ontinues to suffer from micro ial colonization.[64] These can lead to a loss of energy in a system and overall loss of products. costing the healthcare system over $5 illion in additional healthcare expenses. dairy is suscepti le to iofilm formation and contamination. It not only induces iofilm formation. resulting in the uildup of acteria. in part.[62] In produce. and this response can kill acteria y damaging their DNA.[62] This pro lem is also found in ready to eat foods ecause the foods go through limited cleaning procedures efore consumptio n [62] Due to the perisha ility of dairy products and limitations in cleaning pr ocedures. Competent S.[60] Biofilms in medicine[edit] The rapidly expanding worldwide industry for iomedical devices and tissue engin eering related products is already at $180 illion per year. pneumoniae to survive the defenses of the host.[62] Bacteria can survive long periods of time in water.[61] If an infection develops a iofilm.[62] Biofi lms have een connected to a out 80% of acterial infections in the United State s.[62][64] The acteria can spoil the products more re adily and contaminated products pose a health risk to consumers.[63] The uildup of iofilms can affect th e heat flow across a surface and increase surface corrosion and frictional resis tance of fluids. Streptococcus pneumoniae[edit] S. causing iofilm formation on plants or in the processing equipment. As the act eria changes. When S. it ecomes more resistant to anti iotics and the odys own host d efenses.[65] Large amounts of salmonella contamination can e found in the poultry processing industry as a out 50% of salmonella strains can produce iof ilms on poultry farms. the host’s polymorphonuclear leukocytes produce an oxidative urst to defend agai nst the invading acteria.[59] Formation of a iofilm depends on compe tence stimulating peptide (CSP). One acteria th at can e found in various industries and is a major cause of food orne disease is Salmonella.[62]Salmonella increases the risk of food orne illnesses w hen the poultry products are not cleaned and cooked correctly.[58] In particular. iofilms resist sanitization and allow acter ia to spread across the produce. mi cro ial infections can develop on all medical devices and tissue engineering con structs. from 1996 to 2010 the Center for Disease Con trol and Prevention estimated 48 million food orne illnesses per year. p neumonia grows in iofilms.[62] During the washing process.

[65] New forms of cleaning procedures are eing tested in order to reduce iofilm for mation in these processes which will lead to safer and more productive food proc essing industries.[64] .[65] Shrimp products are commonly aff ected y salmonella ecause of unhygienic processing and handling techniques[65] The preparation practices of shrimp and other seafood products can allow for a cteria uildup on the products.on the seafood itself as well as in water. These new forms of cleaning procedures also have a profound e ffect on the environment. often releasing toxic gases into the groundwater reser voirs.