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Staphylococcus aureus biofilm on an indwelling catheter A biofilm is an aggregate of microorganisms in which cells adhere to each other on a surface. These adherent cells are frequently embedded within a self-produced matrix of extracellular polymeric substance (EPS). Biofilm EPS, which is also referred to as slime (although not everything described as slime is a biofilm), is a polymeric conglomeration generally composed of extracellular DNA, proteins, and polysaccharides. Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial and hospital settings. 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, nutritional cues, or in some cases, by exposure of planktonic cells to sub-inhibitory concentrations of antibiotics. When a cell switches to the biofilm mode of growth, it undergoes a phenotypic shift in behavior in which large suites of genes are differentially regulated.

Composition of biofilm
Biofilms consist of microorganisms and their self-produced extracellular polymeric substances (EPS). A fully developed biofilm contains many layers including a matrix of EPS with vertical structures, and a conditioning film. Vertical structures of microorganisms sometimes take the form of towers or mushrooms, and are separated by interstitial spaces. Interstitial spaces allow the bulk of the biofilm to easily and rapidly take in nutrients from the surrounding liquid and move byproducts away from the biofilm.

the biofilm grows through a combination of cell division and recruitment. trapped biofilm bacteria form a community that controls the structural complexity of the biofilm Formation of a biofilm begins with the attachment of free-floating microorganisms to a surface. It is during this colonization that the cells are able to communicate via quorum sensing using such products as AHL. The development of a biofilm may allow for an aggregate cell colony (or colonies) to be increasingly antibiotic resistant. cell-cell growth population growth / reproduction 3. initial adsorption to surface 2. The final stage of biofilm formation is known as development. production of an extracellular polysaccharide substances / irreversible adhesion 4. . Once colonization has begun. These first colonists adhere to the surface initially through weak. they can anchor themselves more permanently using cell adhesion structures such as pili. The first colonists facilitate the arrival of other cells by providing more diverse adhesion sites and beginning to build the matrix that holds the biofilm together. If the colonists are not immediately separated from the surface. Some species are not able to attach to a surface on their own but are often able to anchor themselves to the matrix or directly to earlier colonists. reversible adhesion via van der Waals forces. and is the stage in which the biofilm is established and may only change in shape and size.Formation 1.

CSLM. which have been shown to cross-link with the polymer strands and provide greater binding force in a developed biofilm. lipids and even humic substances). phosphate promotes the EPS synthesis. These multispecies micro consortia can result from association between metabolically co-operative organisms. EPS may vary in chemical and physical properties but it primarily consists of polysaccharides. It restricts the diffusion of compounds from surroundings into the biofilm. The presence of uronic acids D-glucuronic.Biofilm structure Observation of the biofilm without disrupting the community. The amount of EPS produced by different organisms may vary and the amount of EPS increases with the age of biofilm. Reported 9. Biofilm is composed primarily of micro colonies of different species of microbial cells (+15% by volume) and of matrix material (+85%). EPS may associate with metal ions. This property helps in the association of divalent cations such as calcium and magnesium. which allows the visualization of fully hydrated sample. The EPS matrix also has the potential to physically prevent the access of certain antimicrobial agents into the biofilm by acting as an anion exchanger. DNA. EPS production is known to be affected by nutrient status of the growth medium. enhancing the nutrient availability as 11 well as removal of potentially toxic metabolites. 4- linked hexose residues. 8 EPS has also been reported sequester. Role of Biofilm in Microbial Communities Biofilm formation is prevailing in bacterial life style. Micro colonies in biofilm quite often consist of different microbial communities. This effect appears to be more pronounced with the antibiotics that are hydrophilic and positively charged such as 7 aminoglycosides 7. Their close proximity facilitates interspecies substrate . Nutrient Availability The water channel provides effective means of exchanging nutrient and metabolites with the bulk aqueous phase. however. D-galactouronic and mannuronic) or ketal linked pyruvate confers the anionic properties. Here.or 1. we have described why biofilm strategy 2 has been adopted by so many microbes. provide cations protection and toxins from variety and of environmental stresses such as pH shift. These characteristics largely depend on the nature of both the agent and the EPS matrix. 10 metal to ions. UV radiation. Backbone of EPS contains 1. divalent cautions and macro molecules (proteins. osmotic shock and desiccation. excess available carbon. Protection from Environment EPS of biofilm provides certain degree of shelter and homeostasis to the bacteria residing in biofilm. limitation of nitrogen. potassium. Some of the polysaccharides are neutral or polyanionic. 3. EPS plays various roles in structure and function of different biofilm communities. has revealed the elaborate 3-dimensional structure of biofilm.

flagellar synthesis decreases. for the establishment of syntrophic case of Syntrophism is a special symbiosis in which two metabolically distinct bacteria depend on each other to utilize certain substrates. antibiotics. 14 will reinforce the biofilm structure biofilm structure and 14. Researchers have noted that the pulmonary isolates (Pseudomonas aeruginosa) are mucoid due to the synthesis of large amounts of alginate. which are then utilized as substrate by acetogenic bacteria. With regard to methanogenic. Methanogen obtains energy by converting acetate. Fermentive bacteria initiate the catabolism producing acids and alcohols. In and removal and distribution of metabolic products. the bacterium must differentiate into biofilm-associated cells by repressing the synthesis of flagellum that destabilizes the biofilm and producing exopolysaccharides that 13. . to become an effective member of a biofilm community. typically for energy requirements. This reveals that when synthesis of the exopolysaccharides and alginate are increased in the biofilm-associated cells. degradation of complex organic matter into methane and carbon dioxide during anaerobic digestion requires interaction of at least three bacteria. The mechanisms responsible for the resistance to these antimicrobial agents. Transcription of algC gene involved in the production of alginate is increased approximately fourfold in biofilm associated cells as compared to planktonic cells. 15 the nature of physiological attributes of biofilm organisms confer inherent resistance to the antimicrobial agents. Acquisition of new genetic trait gives chances to the microbial communities to transcribe the necessary genes to become the active member of the biofilm communities. disinfectants or germicides are described below. Acquisition of New Genetic Trait Horizontal gene transfer is important for the evolution and genetic diversity of natural microbial community. This is due to transcription of different genes by biofilm forming communities and the phenotypic characters are the expression of a particular genotypic character. For example. 12. Thus. by mutational analysis researchers have shown that alginate synthesis is positively regulated by sigma factors whereas sigma factor negatively regulates the synthesis of flagellum. carbon dioxide and hydrogen to methane. Biofilm provides an ideal environment relationship. Syntrophism has been well degradation studied 11.

All photomicrographs are shown to same scale. 4. may play a role in biofilm dispersal. such as dispersin B and deoxyribonuclease. aeruginosa biofilm. Dispersal enables biofilms to spread and colonize new surfaces. Enzymes that degrade the biofilm extracellular matrix. There are five stages of biofilm development (see illustration at right): 1.Development Each stage of development in the diagram is paired with a photomicrograph of a developing P. 3. [10] Recent evidence has shown that a fatty acid messenger. 2. . Initial attachment: Irreversible attachment: Maturation I: Maturation II: Dispersion: Dispersal Dispersal of cells from the biofilm colony is an essential stage of the biofilm life cycle. cis-2-decenoic acid. is capable of inducing dispersion and inhibiting growth of biofilm colonies. Nitric oxide has the potential for the treatment of patients that suffer from chronic infections caused by biofilms. Secreted by Pseudomonas aeruginosa. Biofilm matrix degrading enzymes may be useful as anti-biofilm agents. this compound induces cyclo heteromorphic cells in several species of bacteria and the yeast Candida albicans. Nitric oxide has also been shown to trigger the dispersal of biofilms of several bacteria species at sub-toxic concentrations. 5.

Bacteria living in a biofilm usually have significantly different properties from freefloating bacteria of the same species. protozoa. the biofilm does have greater resistance to antimicrobials. This resistance to antibiotics in both stationary phase cells and biofilms may be due to the presence of persister cells. Some biofilms have been found to contain water channels that help distribute nutrients and signalling molecules. the biofilm form of Pseudomonas aeruginosa has no greater resistance to antimicrobials than do stationary-phase planktonic cells. as the dense and protected environment of the film allows them to cooperate and interact in various ways. Lateral gene transfer is greatly facilitated in biofilms and leads to a more stable biofilm structure. Given sufficient resources for growth. a biofilm will quickly grow to be macroscopic (visible to the naked eye). some organisms will form single-species films under certain conditions. This matrix is strong enough that under certain conditions. fungi and algae. although when the biofilm is compared to logarithmic phase planktonic cells. . biofilms are not always less susceptible to antibiotics. biofilms can become fossilized. as the dense extracellular matrix and the outer layer of cells protect the interior of the community. bacteria. e. EPS is an abbreviation for either extracellular polymeric substance or exopolysaccharide. although they can form as floating mats on liquid surfaces and also on the surface of leaves. However. In some cases antibiotic resistance can be increased a thousandfold.Properties Biofilms are usually found on solid substrates submerged in or exposed to an aqueous solution. each group performs specialized metabolic functions. Biofilms can contain many different types of microorganism. One benefit of this environment is increased resistance to detergents and antibiotics. particularly in high humidity climates. For instance. This matrix protects the cells within it and facilitates communication among them through biochemical signals. However.g. archaea. Extracellular matrix The biofilm is held together and protected by a matrix of excreted polymeric compounds called EPS.

Pre-requisite for biofilm formation 2. Once a material is exposed to water. 2. Binds the bacteria together to form the biofilm  Sticking Efficiency Equation = sticking efficiency = number of cells adsorbed onto substratum = number of cells transported to substratum . organic molecules begin to adsorb to its surface.Biofilm adhesion How a biofilm adheres to the surface of a material Polymicrobic biofilm epifluorescence  Adsorption: the interphase accumulation of cells from the bulk liquid directly on the substratum (surface of the material) 1. Called the conditioning film: mainly composed of glycoproteins (subject to high turnover rate (not static) EPS (extracellular polymeric substances) 1.

In order to understand the process of attachment one must first examine the properties of both the substratum and the cell surface  Substratum: can be either very hydrophobic (Teflon) or hydrophilic (glass) 1. in order to prevent bacteria from forming a biofilm the substratum should be incredibly smooth. The reason that these are important is because a cell. these appendages enable the cell to remain attached until better/more capable attachment mechanisms are set Inhibiting protein adsorption/ biofilm adhesion Judging from what has been uncovered through the processes of Adsorption and Attachment. Attachment: the acquisition of cells from the bulk liquid by an existing biofilm 1. The Methods section will cover more detail in this regard. . once drawn to the substratum must combat the repulsive forces common for all materials. Another method could be to chemically coat the substratum in order to prevent the conditioning layer and the EPS from forming. or glycocalyx may impact rate of microbial attachment 1. This will make it difficult for the cells to attach to the surface. fimbriae. Rougher and more hydrophobic materials will develop biofilms faster  Cell properties: flagella. pili.

 Polymer modification Dispersive forces of grafted polymer chains can prevent bacterial adhesion to a surface . However. Typically. a process in which metal ions interfere with the growth and function of bacteria. after which leaching of the antimicrobial agent reduces the effectiveness of the coating. wine. Several in vitro studies have confirmed the effectiveness of silver at preventing infection. They prevent biofilm formation by interfering with the attachment and expansion of immature biofilms. and vinegar in silver bottles to keep them from spoiling. both in coating form and as nanoparticles dispersed in a polymer matrix. For this reason. biocides. silver coatings are commonly used on devices such as catheters. Antibiotics. its use can be traced to the Phoenicians. Despite this. some fear that silver may have a similarly toxic effect on human tissue. Considering the mechanism by which silver interferes with bacterial cell function. There has been renewed interest in silver coatings for antimicrobial purposes. who would store their water.Methods Chemical Antimicrobial coatings Chemical modifications are the main strategy for biofilm prevention on indwelling medical devices. The antimicrobial property of silver is known as an Oligodynamic effect. there has been limited use of silver coatings in vivo. and ion coatings are commonly used chemical methods of biofilm prevention. The medical uses of silver and silver ions have been known for some time. these coatings are effective only for a short time period (about 1 week). concerns remain over the use of silver in vivo.

an antimicrobial agent. The concept is similar to that of Steric Stabilization of colloids. These chains are anchored to the device surface by covalent bonds. Dispersion forces between the polymer chains and the bacterial cells prevent bacteria from binding to the surface and initiating biofilm growth. The solubility of these polymers stems from the high conformational entropy of polymer chains in solution. Adherence is thermodynamically . producing non-leaching. coli. the polymer will be soluble. aeruginosa bacteria. One in vitro study found that when N-alkylpyridinium bromide. and T is temperature in Kelvins.To avoid the undesirable effects of leaching. R is the ideal gas constant. and P. epidermidis. The Χ (Chi) parameter is used to determine whether a polymer will be soluble in a given solution. is the molar volume of the solution (assuming ). Χ is given by the equation: where and are the cohesive energy densities of the polymer and solvent. flexible polymeric chains. Mechanical  Hydrophobicity Contact angle of a liquid droplet wetted to a rigid solid surface. contact-killing surfaces. If 0 < < 2. respectively. was attached to a poly (4vinyl-N-hexylpyridine). antimicrobial agents can be immobilized on device surfaces using long. the polymer was capable of inactivating ≥99% of S. The ability of bacteria to adhere to a surface and begin the formation of a biofilm is determined in part by the enthalpy of adhesion of the surface. E. Polymer chains are grafting to a surface via covalent bonding or adsorption.

R is the ratio of actual surface area over projected surface area. respectively. solid–vapor interfaces. It is thus desirable to maintain a smooth surface on any products that may come in contact with bacteria. The roughness of a surface can affect the hydrophobicity or hydrophilicity of the contacting substance. Using this equation Surface roughness Wenzel model Surface roughness has canned also affect biofilm adhesion. and the can be determined. and are the interfacial energies of the solid–liquid. .favored if the free enthalpy of adhesion is negative and decreases with increasing free enthalpy values. Studies have shown that there is a threshold value of surface roughness (Ra = 2 microns) below which biofilm adhesion will reduce no further. while smooth surfaces are less susceptible to biofilm adhesion. which in turn affects its ability to adhere. high-energy surfaces are more conducive to biofilm formation and maturation. The Wetzel equation predicts that a hydrophilic surface will have a lower . Young's Equation can be used to determine whether if adhesion is favorable or unfavorable: Where . The free energy of adhesion can be determined by measuring the contact angles of the substances in question. thus making it easier for bacteria to adhere. the liquid–vapor. The Wetzel equation can be used to estimate the observed contact angle: Where the apparent contact angle and R is the roughness parameter of the surface. Rough.

Ozone cleaves extracellular polysaccharides re-dissolving the bacteria  Low-energy surface acoustic waves This technique uses low-energy waves produced from a battery powered device. Based on the principles of electrostatics charged particles will repel other particles of like charge.  Techniques Due to high interest in preventing biofilm formation in the recent years a large number of techniques have been studied to find a good solution. Positively charged polycationic chains enable the molecule to stretch out and generate bactericidal activity. . This technique has been tested on white rabbits and guinea pigs. in this case a catheter. The hydrophobicity and the charge of polymeric chains can be controlled by using several backbone compounds and antimicrobial agents. The following section summarizes just a few of the paths being examined in the field but many more techniques outside of these are being developed as well. Surface charge Modification of the surface charge of polymers has also proven to be an effective means of biofilm prevention. creating horizontal waves that prevent the adhesion of planktonic bacteria to surfaces. The device delivers periodic rectangular pulses through an actuator holding a thin piezo plate. The results showed a lowered biofilm growth. The waves spread to the surface.

a group of bacterial colonies on a surface. Ozone targets extracellular polysaccharides. The tubing it ran through was tested weekly for bacterial colonies. especially catheters. discharge techniques Surfaces with negative surface charge lessened formation. Ozone is so effective because it is a very strong oxidant and it encounters biofilms in much larger concentrations than most disinfectants like chlorine. The following table is a quick summary of a few surface modification techniques that have been studied. Ozonation Biofilms form as a way of survival for bacteria in aqueous situations. (dimethylamino)ethyl and silicone rubber Silicone rubber had almost no biofilm methacrylate]) present. Water purification methods are being scrutinized here because it is in this state that contamination is thought to occur and biofilms are formed.  Surface modification Surface modifications have been a highly studied technique for biofilm prevention.  Water purification When this technique was studied two purification methods were used to treat water. The highly purified water showed a sharp decrease in bacteria colony adherence. These techniques have been the focus of many biomedical studies aiming to reduce harmful biofilm formation on medical devices.Grafted to polyethylene Biofilm formations were reduced. The other was a double reverse osmosis technique with electric deionization which was continuously disinfected with UV light and disinfected weekly with ozone. After ten days a biofilm layer was completely established. The first was a typical reverse osmosis technique used for pure water. Technique Silver ions Method of action Results Eluted from sol-gel Silver ions initially impeded biofilm coating growth. and cleaves them. Many methods have been tested and a variety of results have been recorded. The ozone cuts through the skeleton of the biofilm at a rapid pace thus dissolving it back to harmless microscopic fragments. pDMAEMA (poly[2. This technique has been employed mainly in the spa and pool industry as a way to purify water. Polyurethane Polymer surface Changes in surface roughness showed modified through glow no effect on biofilm formation. .

K. These organisms may originate from the skin of patient. Raad and Sherertz also showed that catheters in place for less than 10 d tended to have more extensive biofilm formation on the external surface of the catheter. tap water to which entry ports are exposed or other sources in the environment. Aeruginosa. researchers have attempted quantification of biofilm using sonication plus vortexing of catheter . epidermidis. medications. Colonization and biofilm formation may occur within 3 d of atherization. S. depending on the device and its duration of action. In this procedure. the distal tip of catheter is removed aseptically and rolled over the surface of a non-selective medium. Biofilm on central venous catheters have routinely been detected by a semiquantative procedure termed the roll plate catheter biofilm.albicans. During long term catheterization there would be more formation of a biofilm on the inner lumen of catheters. These organisms originate from patient's skin microflora. most commonly isolated from Catheters may be inserted for administration of fluid. Therefore. or health-care workers. are S. Biofilms have been reported to be universally present on central venous catheters using SEM and TEM and may be associated with either the outside of the catheter or inner lumen. nutritional solution. Microorganisms commonly associated with biofilm on indwelling devices are shown in Table 2.Applications of Biofilm Biofilm and Devices Associated Infection Biofilm on indwelling medical devices may be composed of Gram-positive or Gramnegative microorganisms. C. aureus. The organisms. exogenous microflora from health-care personnel. They gain access to the catheter by migration externally from skin along the exterior catheter surface or internally from the catheter port. etc technique. and hemodynamic monitoring. The roll plate technique has the limitation such as low diagnostic sensitivity and low predictive value for catheter-related infection. blood products. Central Venous Catheter Biofilm All the indwelling central venous catheters are colonized by microorganisms embedded in a biofilm matrix. Pneumoniae. P. Biofilms may be composed of single species or multiple species.

urinary catheter Artificial voice prosthesis. DNA intercalating agent Negative staining. prosthetic heart valves. Coagulase -negative Staphylococci . prosthetic heart valve. central venous catheter. intra uterine devices. ficols Stains DNA and RNA Indicates esterase activity Stains neutral lipid and phospholipids RNA. may be conjugated to antibodies. prosthetic heart valve Artificial hip prosthesis. Intra uterine devices. urinary catheter Artificial hip prosthesis. urinary catheter Causative organism Coagulase -negative Staphylococci Klebsiella pneumoniae Pseudomonas aeuginosa Candida albicans Staphylococcus aureus Enterococcus spp. lectins. may be conjugated to antibodies. dextrans. intra uterine devices Artificial hip prosthesis. pH indicator Bind to proteins. central venous catheter. central venous catheter Central venous catheter. lectins and dextran Calcium indicator PH indicator Stains DNA Table 2 — Microorganisms associated with biofilm on indwelling devices Medical devices Urinary catheter. central venous catheter. may be conjugated to antibodies. lectins and dextran Bind to proteins.Table 1 — Common fluorescent probes used in biofilm studies Name FITC Acridine orange Fluoroscein diacetate Nile red Propidium iodide Fluoroscein TRITC RITC Fluo-3 NCECF Name Excitation\emmision 490\520 490\530 495\520 450\530 530\615 490\520 541/572 575/ 595 506/ 526 500/ 530or 620 362/ 470 Applications Binds to proteins.

In the open system. enterococci and Candida spp. catheter quickly gets contaminated and develops urinary tract infection (UTI) within 4 d. These mineral containing biofilms. which is used to bond cephalosporin. greater the tendency of these organisms to develop biofilm and result in UTI. in closed system. Divalent cations (calcium and magnesium). Only 10 to 20% of patients undergoing short-term catheterization (up to 7 d) but essentially all the patients undergoing long-term catheterization (more than 30 d) get infected with UTI . bladder irrigation. Pneumoniae and other Gram-negative bacteria. The higher pH responsible for biofilm-urine interface results in precipitation of minerals such as struvite and hydroxyapatite. Catheters may be pen or closed systems. will less likely to develop biofilm than untreated catheter. may completely block the inner lumen. Likely with chlorhexidine impregnated and with Catheter. In open system. Other research group has found that catheter impregnated with minocycline and rifampicin commonly develops on the tissue surrounding the prosthesis. Aureus. Antimicrobial agents are usually applied during valve replacement and whenever patient has dental work to prevent the initial attachment by killing the microorganisms introduced into the blood stream were less to be colonized than impregnated sulfadiazine. Aeruginosa. Several strategies have been attempted to control the urinary catheter biofilm such as antimicrobials. On the other hand. Streptococcus spp. diptheroids. . and antimicrobial agents in collection bags. Urinary Catheters Urinary catheters are tubular latex or silicone devices that are inserted through urethra into the bladder to measure the urine output and collect urine during surgery. Patients using closed system are much less susceptible to UTI. and impregnation of Catheter with antimicrobial agents such as silver oxide or systemic antibiotics. the catheter drains into an open collection center. S. Some organisms of these biofilms produce urease. The longer the urinary catheter remains in place. Proteus mirabilis. which form encrustations. Epidermidis.Examples Several studies have examined the effect of various types of antimicrobial treatment in controlling the biofilm. The primary microorganisms responsible for this condition are S. Enterococcus faecalis. epidermidis. increase the urinary pH and ionic strength. P. which hydrolyzes the urea to ammonium hydroxide. Researchers have found that addition of sodium metabisulfite to dextrose heparin flush of left arterial catheter eliminated microbial colonization of this catheter. E. coli. K. Gram-negative bacilli. the catheter empties into a securely fastened plastic bag. which results in enhancement of bacterial attachment. Contact Lenses Contact lenses have been classified as soft contact lenses and hard contact lenses according to materials those silver cationic Surfactant. The organisms commonly contaminating these devices and developing biofilms are S.

The degree of attachment to the lenses depends on the nature of substrate. epidermidis and species of Proteus. design. IUDs removed from women with pelvic inflammatory disease may also contain streptococci. and frequency of disposal. the lens case has been implicated as the primary source for contamination. Aureus. Organisms mainly adhering to the contact lenses are P. water content. etc. Fibrin at the Microorganisms suture also site have and on the device. wear schedule. and S. Longest raised mat area is about half a meter long. In fact. coli. Microorganisms readily adhere to the surface of both types of lenses. Biofilm in Yellowstone National Park. Aeruginosa using SEM. coli and some anaerobic bacteria. Biofilms have been observed on the lenses removed from a patient with keratitis caused by P. Epidermidis. S. etc. aureus and species of Corynebacterium. Intrauterine Devices The intrauterine devices (IUDs) have a tail that facilitates locating the device for removal and it is composed of a plastic monofilament surrounded by a nylon sheath. Thermophilic bacteria in the outflow of Mickey Hot Springs. S. . The tail portion of the IUDs may be a primary source of contamination. Enterococcus. Candida albicans. S. approximately 20 mm thick. type of bacterial strain. Organisms which contaminate the IUDs are actobacillus plantarum. aureus. polymer composition. Greater tendency to colonize these locations. Serratia. S. etc.Prosthetic Heart Valves Mechanical valves and bioprostheses are being currently used as prosthetic heart valves. Biofilms have also been found to develop on contact lenses kept in storage cases. Candida. aeruginosa. E. The surgical implantation of the prosthetic valve results in tissue damage. E. electrolyte concentration. Oregon. leading to the accumulation of platelets and of construction.

What we regard as clean water is a waste material to these microcellular organisms since they are unable to extract any further nutrition from the purified water. Biofilms in cooling. in particular by a remarkable recently-discovered group of specialists. where they may cause tooth decay and gum disease. Stromatolites include some of the most ancient records of life on Earth. Time in dry dock for refitting and repainting reduces the productivity of shipping assets. including pathogens and other microorganisms. Such fouling can reduce maximum vessel speed by up to 20%. Bacterial adhesion to boat hulls serves as the foundation for biofouling of seagoing vessels. at least 20% of corrosion is caused by microorganisms that are attached to the metal subsurface (i. binding and cementation of sedimentary grains by microbial biofilms.or heating-water systems are known to reduce heat transfer.Biofilms are ubiquitous. Nearly every species of microorganism. not only bacteria and archaea. especially of cyanobacteria. bacteria are mainly responsible for removal of organic matter (BOD). biofilms are important components of food chains in rivers and streams and are grazed by the aquatic invertebrates upon which many fish feed. and the useful life of ships is also reduced due to corrosion and mechanical removal (scraping) of marine organisms from ships' hulls.e. Biofilms on floors and counters can make sanitation difficult in food preparation areas. In the human environment.[18] Biofilms in marine engineering systems. however. such as pipelines of the offshore oil and gas industry. Slow sand filters rely on biofilm development in the same way to filter surface water from lake. Biofilms can help eliminate petroleum oil from contaminated oceans or marine systems. and are still forming today. the extremely hot. for example. it is easier for other marine organisms such as barnacles to attach. briny waters of hot springs ranging from very acidic to very alkaline. The oil is eliminated by the hydrocarbon-degrading activities of microbial communities.           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 can also be harnessed for constructive purposes. For example. Biofilms are present on the teeth of most animals as dental plaque. which extract and digest organic compounds. have mechanisms by which they can adhere to surfaces and to each other. the so-called hydrocarbonoclastic bacteria (HCB). microbially-influenced corrosion). can lead to substantial corrosion problems. Biofilms will form on virtually every non-shedding surface in a non-sterile aqueous (or very humid) environment. spring or river sources for drinking purposes. Stromatolites are layered accretionary structures formed in shallow water by the trapping. biofilms can grow in showers very easily since they provide a moist and warm environment for the biofilm to thrive. Biofilms can grow in the most extreme environments: from. Corrosion is mainly due to abiotic factors. many sewage treatment plants include a treatment stage in which waste water passes over biofilms grown on filters. . to frozen glaciers. prolonging voyages and consuming fuel. Once a film of bacteria forms. while protozoa and rotifers are mainly responsible for removal of suspended solids (SS). In such biofilms. Biofilms can form inside water and sewage pipes and cause clogging and corrosion. In fact.

middle-ear infections. They can either contribute to crop disease or.New staining techniques are being developed to differentiate bacterial cells growing in living animals. infections in cystic fibrosis. contribute to a better clinical management of chronically infected patients and should lead to the . The species of bacteria from interoperative cultures did not correspond to the bacteria species in the biofilm on the respective patient's tissue.[21] Examples of crop diseases related to biofilms include Citrus Canker. Pierce's Disease of grapes. catheter infections.[24] gingivitis. Pseudomonas aeruginosa biofilms The achievements of medical care in industrialized societies are markedly impaired due to chronic opportunistic infections that have become increasingly apparent in immunocompromised patients and the aging population.g. as in the case of nitrogen-fixing Rhizobium on roots.  Biofilms are found on the surface of and inside plants.[23] Infectious processes in which biofilms have been implicated include common problems such as urinary tract infections. e. aeruginosa pathogenicity. It has recently been shown that biofilms are present on the removed tissue of 80% of patients undergoing surgery for chronic sinusitis. and Bacterial Spot of plants such as peppers and tomatoes. the cultures were negative though the bacteria were present. exist symbiotically with the plant. from tissues with allergyinflammations. formation of dental plaque. Chronic infections remain a major challenge for the medical profession and are of great economic relevance because traditional antibiotic therapy is usually not sufficient to eradicate these infections. More recently it has been noted that bacterial biofilms may impair cutaneous wound healing and reduce topical antibacterial efficiency in healing or treating infected skin wounds. by one estimate 80% of all infections. prosthetic cardiac valves and intrauterine devices. In other words. In this context the elucidation of the molecular mechanisms responsible for the switch from planktonic growth to a biofilm phenotype and the role of inter-bacterial communication in persistent disease should provide new insights in P. Biofilms are used in microbial fuel cells (MFCs) to generate electricity from a variety of starting materials. One major reason for persistence seems to be the capability of the bacteria to grow within biofilms that protects them from adverse environmental factors. unlike the controls without biofilms who had normal cilia and goblet cell morphology. Pseudomonas aeruginosa is not only an important opportunistic pathogen and causative agent of emerging nosocomial infections but can also be considered a model organism for the study of diverse bacterial mechanisms that contribute to bacterial persistence. Biofilms and infectious diseases Biofilms have been found to be involved in a wide variety of microbial infections in the body. and infections of permanent indwelling devices such as joint prostheses and valves.[29] Biofilms were also found on samples from two of 10 healthy controls mentioned. and less common but more lethal processes such as endocarditis. The patients with biofilms were shown to have been denuded of cilia and goblet cells. including complex organic waste and renewable biomass.[24] coating contact lenses. Biofilms can also be formed on the inert surfaces of implanted devices such as catheters.

Neisseria gonorrhoeae biofilms Neisseria gonorrhoeae is an exclusive human pathogen. in which they are protected against disinfectants.[33] Dental plaque Dental plaque is the material that adheres to the teeth and consists of bacterial cells (mainly Streptococcus mutans and Streptococcus sanguinis). . or maintained. There is evidence for formation of gonococcal biofilms on human cervical epithelial cells during natural disease and that outer membrane blebbing by the gonococcus is crucial in biofilm formation over human cervical epithelial cells. type IV pili. Understanding the molecular mechanisms of biofilm formation has facilitated the exploration of novel strategies to control bacterial biofilms. Biofilm development can be divided into several key steps including attachment.[36] Molecular genetics Technological progress in microscopy. Recent studies have demonstrated that it utilizes two distinct mechanisms for entry into human urethral and cervical epithelial cells involving different bacterial surface ligands and host receptors. micro colony formation. This accumulation of microorganisms subjects the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease. DNA and exopolysaccharides. In addition it has been demonstrated that the gonococcus can form biofilms on glass surfaces and over human cells. Workers in cooling towers.identification of new drug targets for the development of alternative anti-infective treatment strategies. constructed. dental implants. The bacterial strains identification of the biofilms isolated from the dental plaque or from the biofilms attached to the surfaces of some dental alloys. salivary polymers and bacterial extracellular products.The rapid progress in biofilm research has also unveiled several genetic regulation mechanisms implicated in biofilm regulation such as quorum sensing and the novel secondary messenger cyclic-di-GMP. molecular genetics and genome analysis has significantly advanced our understanding of the structural and molecular aspects of biofilms. restorative and cement materials play an essential role concerning the biofilms establishment dynamics towards the physical-chemical properties of the materials which biofilms are attached to. especially of extensively studied model organisms such as Pseudomonas aeruginosa. Legionellosis Legionella bacteria are known to grow under certain conditions in biofilms. persons working in air conditioned rooms and people taking a shower are exposed to Legionella by inhalation when the systems are not well designed. Plaque is a biofilm on the surfaces of the teeth. impression materials. biofilm maturation and dispersion. and in each step bacteria may recruit different components and molecules including flagellae.

The resistance of microbes residing in the biofilms towards various types of antimicrobial agents poses a serious threat the pharmaceutical industries. These systems should closely simulate the in vivo or in situ conditions for each device. Therefore. device design modifications or different media formulations. effective treatment strategies for complete eradication of biofilms and complete understanding of make the two biofilm phenotype different Counterparts. it is recommended to prevent their formation rather than treatment. . Model systems should be developed and used to study biofilm processes on various indwelling medical devices. This system design could be used to investigate and compare various biofilm control treatments. Biofilms are difficult to remove from blood processing surfaces and evironments due to the production associated equipment of EPS materials and the with and cleaning processing Complex difficulties processing environments. Further study on the biofilms include effective control strategies to prevent the formation of biofilms. Microorganism’s wet so on which from planktonic have been surfaces observed to aggregate and grow into micro colonies form 3-dimensional structures. resulting in a complex biofilm.Conclusions Residual throughout the distribution the importance from a public health perspective is the role of biofilm in antimicrobial drug resistance.

 Watnick P & Kolter R. 8 (2002) 881-890.  Kumar C G. city of microbes. J  Bacteriol.REFERENCE  www. .wikipedia. Biofilm. 1 (2001) 6-7.  Deibel  Donlan R M. J Food Safety. 182 (2000) 2675-2679. Emerg  Infect Dis. 42 (1998) 9-27.  www. Int J Food Microbiol. Biofilms: Microbial life on surfaces. Significance of microbial biofilm in food  Industry: A

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