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Grobe, K. Role of dose concentraion in biocide efficacy against Pseudomonas aeruginosa. Allison, D. Ruiz, B. Extracellular products as mediators of the formation and detachment of Pseudomonas fluorescens biofilms. Gov't Research Support, U. Gov't, Non-P. The exopolysaccharides EPS and water channels formation occur, facilitating nutrient supply which leads to the maturation of the biofilms. Microbial cells within biofilms have shown 10— times more antibiotics resistance than the planktonic cells [ 79 ].
Biofilm is formed in diverse environmental niches, including freshwater rivers, rocks, deep-sea vents and hydrothermal hot springs. Biofilm-related infections can be broadly divided into two types. The biofilms may be formed on the abiotic surfaces, especially infections associated with indwelling medical devices [ 34 ] and native biofilm infections of host tissue [ 21 ]. Urinary tract and bloodstream infections can be caused by the biofilm initially formed on medical implants, such as heart valves, catheters, contact lenses, joint prostheses, intrauterine devices and dental unit.
These infections can only be treated by removal of the implants which not only increasing the cost of the treatment but also it becomes problematic for patients [ 26 ]. Host tissue related biofilm infections are often chronic, including chronic lung infections of cystic fibrosis patients, chronic osteomyelitis, chronic prostatitis, chronic rhinosinusitis, chronic otitis media, chronic wounds, recurrent urinary tract infection, endocarditis, periodontitis and dental caries [ 21 ].
Some of the major biofilm associated infections causing human diseases are listed in Table 1. Antibiotic resistance of bacteria in the biofilm communities contributes to the chronic infections. Resistance mechanisms of biofilm communities are not similar as the planktonic ones such as target site mutations, lower cell permeability, efflux pumps, drug modifying enzymes and drug neutralizing proteins [ 14 , 70 , 72 , 76 , 77 , 96 , , , , ].
A panel of studies suggested that conventional antibiotic resistance mechanisms are unable to explain the various cases of antibiotic-resistant biofilm infections [ 2 , 5 , 19 , ]. It has been reported earlier that repeated exposure of ceftazidime in biofilm-growing Pseudomonas aeruginosa developed the conventional type of intrinsic antibiotic resistance in biofilms infections [ 10 ].
In biofilm communities, antibiotics resistance appears due to various strategies Fig. These mechanisms are the consequences of the multicellular nature of biofilms which leads to the antibiotics resistance of biofilm communities [ 30 ] along with the known conventional resistance mechanisms and makes the failure of treatment strategy.
Multicellular nature of biofilm is the key factor of antibiotics resistance of biofilm communities which is the actual cause of the resistance mechanisms as discussed above. A series of researches exists on formation of biofilm as a multicellular developmental process [ 65 , 98 , ].
Extracellular polymeric substances EPS hold the bacterial cells together and lead to the development of multicellular consortia which makes the heterogenous environment inside the biofilm and initiates the biofilm to function as a multicellular system. Biofilm development is well organized and during its development intercellular and intracellular signaling occurs.
Maturation of the biofilm into complex structures is regulated by the signalling among the cells by the quorum sensing process [ 31 ]. Multicellularity nature of biofilm bacterial communities is responsible for antibiotics resistance; if we can disrupt any step in the formation of multicellular structure of the biofilm than antibiotics efficacy as well as the host defences might be increased which leads to quick treatment of this persistent infection.
On the basis of these observations we can say that multicellular developmental process of the biofilms are important because its insight will open up new targets and approaches for designing new drug molecules against antibiotics resistant microorganisms. Diagrammatic representation of the potential mechanisms of antibiotic resistance in biofilms communities.
Antibiotics resistant state of the biofilm cells lead to a treatment complications in the series of human infections which include biofilm formation on various biological implants such as, heart catheters, urinary catheters, joint implants and replacement of heart valves [ ]. Biofilms pose a threat to the human race because of their persistent nature and plays a major role in certain pathogenic infections [ 40 , 58 , , ]. Studies suggested that role of EPS have been conferring tolerance to aminoglycosides [ 41 , 61 ].
EPS might quench the activity of antibiotics that diffuse through the biofilms via diffusion—reaction inhibition phenomenon, which may chelates the antibiotics by complex formation or degrade through enzymatically based reactions [ 17 , ]. Stationary phase a slow or non-growth phase of the bacterial life cycle and viable-but-nonculturable state VBNC state or a state of dormancy are the ways of survival for bacterial biofilms communities under antibiotics stress [ 20 , 73 ].
Biofilms possesses many cells of stationary phase which have the decreased antibiotics susceptibility to the antibiotics. Many biofilms communities enter into the stationary phase with time which suggested that older biofilms show higher tolerance to antibiotics [ 89 ]. Persisters are another dormancy state of bacterial subpopulation, which have the multidrug tolerance phenotypic rather than genetic variations [ 8 , 47 ].
In stationary state of biofilms communities, persisters might be the prevalent [ 60 ]. One of the antibiotics resistance mechanisms of biofilms communities is the uptake of resistance genes by horizontal gene transfer [ 79 ]. Biofilms provides the compatible conditions for the horizontal gene transfer such as high cell density, increased genetic competence and accumulation of genetic elements or uptake of resistance genes [ 41 ]. Conjugation is the only mechanism of horizontal transfer of resistant genes in biofilms and may confirm the resistance to several antibiotics.
Few studies suggested that conjugation has been shown more efficient in biofilms as compared to planktonic ones [ 66 , 75 , , ]. A penal of studies reported that in vitro mycobacterial biofilms were resistant to antibiotics amikacin and clarithromycin or disinfectants [ 44 , ]. It has been reported earlier that differences between the MIC and minimum biofilm eradication concentration MBEC in 4 species of RGM [ 91 ] and suggested that ciprofloxacin as an effective antibiotic against these biofilms as compared to clarithromycin and amikacin.
Few studies shown the effect of antibiotics in different stages of biofilm development [ 90 , 91 , 92 ] and revealed that at early stage of biofilm development antibiotics treatment was more effective, probably due to the cells which are not completely adapted into biofilm communities.
In an attempt to evaluate mechanisms for these resistance patterns, it has been suggested that permeability of anti-tuberculosis drugs were independent among the mycobacterial species [ ]. Metabolic state and activation of resistance genes like methylases are indispensable for the development of antibiotics resistance in mycobacteria [ 36 , 44 ].
Successful treatment of biofilm-associated infections is troubled due to high antibiotic resistance in these bacterial communities. Classical antibiotics chemotherapy is unable to completely eradicate bacterial cells which are situated in the central region of the biofilm and leads to the emergence of the worsen situation globally.
Therefore to overcome the drug resistance of bacterial biofilm communities; alternative strategies Fig. Diagrammatic representation of the alternative approaches against antibiotic resistant biofilms communities. Naturally produced small molecules by bacterial biofilm communities such as D-amino acids and Polyamine norspermidine; induced the dispersal of mature biofilms which could prevent biofilm formation in S.
These molecules could be used as antibiofilm agent in the biofilm dispersal strategy. Tween 80 is more active against mycobacterial biofilm than NAC because mycobacterial cell wall as well as extracellular matrix possesses high lipid content and suggested that synergistic effect of drugs and anti-biofilm agent may effective in the treatment of infections associated with mycobacterial biofilms communities.
Degradation of the biofilm matrix by biofilm matrix degrading enzymes DNase I, Dispersin B DspB and a-amylase is also another promising antibiofilm strategy. Degradation of biofilm structural component allows the increased penetration of antibiotics which enhances the antibiotics effeciency. Formation of biofilms were controlled by the quorum sensing QS signalling genes and their products.
Halogenated furanone isolated from Delisea pulchra marine algae interrupt the bacterial QS signalling [ 74 ]. Recently, Kaur et al. Attenuation of bacterial QS signalling by ginseng extract, garlic extract, usnic acid and azithromycin possesses inhibitory activity against bacterial and fungal biofilms [ 18 , 51 , ].
Signalling molecule nitric oxide NO disperse the biofilms in P. Most recently our group used the CRISPRi technology to knockdown the luxS gene of QS signalling and fimbriae associated gene fimH for controlling the biofilm mediated infections [ , ]. Methanolic extract of a coral-associated actinomycete helps to reduce biofilm formation of S.
Another natural product, 4-phenylbutanoic acid show high antibiofilm activity against Gram positive and Gram negative bacteria [ 97 ]. Azadiracta indica Neem and Acacia extracts showed antimicrobial effect against S. Limitations of the conventional antibiotic treatments reduced penetration and retention in cell or biofilm were overcome by their nano-formulations which have the ability to cross the biological barrier. Since the last few years, different type of nanoparticles have been used as antimicrobial and antibiofilm metal nanoparticles, organic nanoparticles, green nanoparticles and their combinations [ 9 ].
Kulshrestha et al. It has reported earlier that PDT has sufficiently reduced the clinically-relevant microbes, such as drug resistant Gram-positive and Gram-negative bacteria [ 23 ]. PDT has significant advantages over conventional treatment owing to its ability of selective binding to the membranes of pathogenic cells and the possibility for accurate delivery of light to the affected tissue for the maximal damage of microbes as well as minimal damage of the host [ ].
Recently, our group has shown that PDT could be used to eliminate the biofilms related issues in S. Bacterial antibiotic resistance is also one of the consequences of the bacterial biofilm communities which contribute to the chronic infections. These biofilm communities have few additional resistance mechanisms as compared to the planktonic ones which hamper the treatments option and leads to emergence as well as spreading of the chronic bad bugs. Emergence and spreading of multidrug resistant, extremely drug resistant and total drug resistant strains of M.
In this timeline review we have discussed the mechanisms of antibiotics resistance in biofilms communities and alternative therapeutic options to combat the resistance mediated by chronic bacterial biofilm infections. Alternative approaches, like nanoparticles based antibiotics formulation, novel anti-biofilm agents, CRISPRi gene editing technologies and photodynamic therapy might be the future options to treat the infections caused by multidrug resistant, extremely drug resistant and total drug resistant strains of M.
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Nitric oxide signaling in Pseudomonas aeruginosa biofilms mediates phosphodiesterase activity, decreased cyclic di-GMP levels, and enhanced dispersal.
J Bacteriol. Mycobacterial biofilms: revisiting tuberculosis bacilli in extracellular necrotizing lesions. Microbiol Spectr. Site of action of two ribosomal RNA methylases responsible for resistance to aminoglycoside. J Mol Biol. Beloin C, Ghigo JM. Although only patulin had no effect on the P. Another study showed that the mixture of the quorum controlling compounds with the antibiotic tigecycline increased the susceptibility of S.
Furthermore, the treatment of S. Considering the rising number of antibiotic-resistant pathogens, QS inhibitors can be used as a mixture with the remaining sensitive antibiotics to complement their effects. These molecules mainly act by suppressing the QS system, and their practice with antibiotics leads to effective cure at much lower dosages of the drug than necessary, which may result in reduced therapeutic costs.
These combinations can be beneficial in the cure of chronic infections, such as chronic urinary tract, cystic fibrosis, or prosthetic infections and biofilms are a barrier to antibiotic diffusion in these chronic diseases.
There is an urgent need for new methods in the cure of biofilm-associated infections. For instance, cyclic di-GMP c-di-GMP is a commonly protected prokaryotic second messenger signal molecule necessary for biofilm development [ 58 ]. New inhibitors of diguanylate cyclase enzymes were identified by using in silico screening, and they tested them successfully in vitro. Inhibitors of flow pumps can also be recommended to complement the effect of antimicrobial agent and needed to be tested in vivo.
The choice of antimicrobial agents also seems to be significant because some of them may act as agonists for biofilm formation and some may disrupt it. The usage and dosages of novel antibiotics should be checked and clinically synthesized antibiotics should be tested at impactful concentrations by considering their distribution in biofilms and the detrimental effects of signaling molecules. Other compounds act as key enzymes in the biosynthesis of these signaling molecules and play a role in regulating virulence factor production and biofilm formation.
A ligand-based strategy will allow the identification of new inhibitors in the future. Better usage of the new active molecules can be supported by understanding mechanisms of antimicrobial agents activity as well as the molecular mechanisms associated with biofilm formation and recalcitrance [ 5 ]. Biofilm infections are highly resistant to antibiotics and physical treatments and it is known that there are many strategies that support biofilm antibiotic resistance and tolerance, such as persistent cells, adaptive responses, and limited antibiotic penetration.
It is also known that the underlying mechanisms of antibiotic tolerance and resistance in biofilms have a genetic basis in many cases.
In human diseases, highly organized bacterial cells gradually induce immune responses to form biofilms responsible for chronic infections that lead to tissue damage and permanent pathology.
Therefore, the formation of biofilm is considered a critical concern in health care services. Exploring promising cure methods for biofilm-associated infections is an urgent task. Few innovative and effective antibiotic strategies have been tried, such as dispersion of biofilms, antibiotic combinations with quorum sensing inhibitors, and a mixture of all these new techniques. Although the mentioned anti-biofilm strategies are important research areas, they are still in infancy and have not undergone clinical research and entered the commercial market.
We hope that new anti-biofilm molecules based on finding universal substances that do not harm cells and synergistic with commonly used antibiotics will be available in the near future. Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution 3. Help us write another book on this subject and reach those readers. Login to your personal dashboard for more detailed statistics on your publications. Edited by Sadik Dincer.
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Downloaded: Abstract Biofilms can be found on several living and nonliving surfaces, which are formed by a group of microorganisms, complex assembly of proteins, polysaccharides, and DNAs in an extracellular polymeric matrix. Keywords biofilm antibiotic resistance bacteria antimicrobial agents. Introduction Bacteria can grow in biofilms on a wide variety of surfaces and attach to inert or alive surfaces, including tissues, industrial surfaces, and artificial devices, such as catheters, intrauterine contraceptive devices, and prosthetic medical devices, implants, cardiac valves, dental materials, and contact lenses [ 1 , 2 ].
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