BIOFILM FORMATION INPATHOGENS BACTERIAProfosser: Dr.ahmad ali PORBABAEIby: ABBAS MOROVVATI
CONTENT Definition of Biofilm History Biofilm information Biofilm information in vitro Where biofilms are found? Advantage of biofilm Biofilm and antibiotic resistance Dis advantage of biofilm Biofilms and infectious diseases References
DEFINITION OF BIOFILM Biofilm: An aggregate of microbes with a distinct architecture. A biofilm is like a tiny city in which microbial cells, each only a micrometer or two long, form towers that can be hundreds of micrometers high. The "streets" between the towers are really fluid-filled channels that bring in nutrients, oxygen and other necessities for live biofilm communities. The biofilm is held together and protected by a matrix of excreted polymeric compounds called EPS .This matrix protects the cells within it and facilitates communication among them through biochemical signals Attachment by pili and etc
BIOFILM Biofilms consist of:1-microbes2-EPS(poly sacarid--glycoprotein)3-water(85-98%) Biofilm can contain many different types of microorganism, e.g. bacteria, archaea, protozoa, fungi and algae; each group performing specialized metabolic functions. Fungal biofilms also frequently contaminate medical devices. They cause chronic vaginal infections and lead to life-threatening systemic infections in people with hobbled immune systems
HOW IS BIOFILM FORMED?Bacteria continue to grow, outer cells provide a physical barrier to protect inner cells surface Planktonic bacteria, in the presence of water, attach to a surface.
BIOFILM INFORMATION IN VITRO 1_batch culture model 2_cotinuous culture model NOTE Study population of archie bacteria use of coenzyme F420 and use of OR NAD and ATP production and so MONOCULTURE:THIS BIOFILM FORMED BY ONE TYPE OF BACTERIA
. WHERE BIOFILMS ARE FOUND . 1. on solid substratums in contact with moisture, 2. on soft tissue surfaces in living organisms 3. at liquid-air interfaces. 4-Biofilms form on the surface of catheter lines and contact lenses. They grow on pacemakers, heart valve replacements, artificial joints and other surgical implants 5-Biofilms grow in hot, acidic pools in Yellowstone National Park (USA) 6-In industrial environments, biofilms can develop on the interiors of pipes
ADVANTAGE OF BIOFILM Bacteria growing in a biofilm are highly resistant to antibiotics, up to 1,000 times more resistant than the same bacteria not growing in a biofilm. Standard antibiotic therapy is often useless and the only recourse may be to remove the contaminated implant. One benefit of this environment is increased resistance to detergents Biofilms can help eliminate petroleum oil from contaminated oceans or marine systems. Bioreactors for water infiltration
BIOFILM AND ANTIBIOTIC RESISTANCE Little or no effect because the bacteria in biofilm are in a different Bacteriostatic phase than most Antibiotics antibiotics target Surface
DIS ADVANTAGE OF BIOFILM In industrial environments, biofilms can develop on the interiors of pipes, which can lead to clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas Infiltration of wast water Oil industrial Attache to hulk and reduce speed of ship Envelope Destruction such as color and epoxi envelopes Reduce of heat transfer in condenser Biological erosion Biofoulding Reduce speed of water in pipe
BIOFILMS AND INFECTIOUS DISEASES Biofilms have been found to be involved in a wide variety of microbial infections in the body, by one estimate 80% of all infections.Infectious processes in which biofilms have been implicated include common problems such as urinary tract infections, catheter infections, middle-ear infections, formation of dental plaque --gingivitis, coating contact lenses, and less common but more lethal processes such as endocarditis, infections in cystic fibrosis, and infections of permanent indwelling devices such as joint prostheses and heart valves-chronic sinusitis-
DENTAL PLAQUE Dental plaque is the material that adheres to the teeth and consists of bacterial cells (mainly Streptococcus mutans and Streptococcus sanguis), salivary polymers and bacterial extracellular products. Plaque is a biofilm on the surfaces of the teeth. This accumulation of microorganisms subject the teeth and gingival tissues to high concentrations of bacterial metabolites which results in dental disease
ROLES FOR FLAGELLAR STATORS IN BIOFILMFORMATION BY PSEUDOMONAS AEROGINOSA pseudomonas aeroginosa caused sistic fibrosis It has only a single flagellum Genom of bacteria encode 2 flagellar stators called MotAB or the MotCD Flagellar stator produce energy
MYCOBACTERIUM AVIUM A biofilm-forming opportunistic human pathogen found in the environment Several strains have been isolated from AIDS patients and others with compromised immune systems
PAST WORK Past work on genes associated with biofilm formation in M. avium has yielded approximately 12 genes that are up-regulated in biofilm formation. Several of these genes are important for glycopeptidolipid (GPL) biosynthesis, while others play a key role in fatty acid metabolism or the citric acid cycle.
CURRENT WORK Using primers for previously identified genes associated with biofilm formation, quantify gene expression M. avium strains MAC A5, MAC 101, and MAC 104 in the presence and absence of three different antibiotics at sub-inhibitory concentrations using Real-Time PCR.
GENES / GENE PRODUCTSBiofilm genes Glycosyl Transferase, essential for the expression of mature GPLs. GuaB2 (IMP dehydrogenase), catalyzes the first reaction in GMP biosynthesis PmmB (Phosphomannose mutase), converts D- Mannose 1-Phosphate TO D-Mannose 6- PhosphateControl gene 16S RNA, not involved in biofilm formation, to be used as a control
TIME COURSE OF BIOFILM FORMATION BYSTAPHYLOCOCCUS AUREUS AND STAPHYLOCOCCUSEPIDERMIDIS MASTITIS ISOLATES Isolation mastitis staphylococcal Loci gene Isolatin with fluorescent in situ hybridisation(FISH) TSB Fixation by paraformaldehyde Change in pentration by ethanol,lysostaphin Hybridation with 16SrRNA Probes :sta and sau
EFFECT OF SHEAR STRESS ON GROWTH ,ADHESION AND BIOFILMFORMATION OF PSEUDOMONAS AEROGINOSA WITHANTIBIOTIC_INDUCED MORPHLOGICAL CHANGES Effect of morphological changes and stress on growth and attachment biofilm formation pseudomonas aeroginosa ATCC27853 Microtitre plate assay Effect of .5 piperacillin tazobactam imipenem meropenem Orbital shaking 250rpm Decrease of attachment and biofilm information Use of sub_mic antibiotic(minimal inhibitory concentration)
BILE SALTS ENHANCE BACTERIAL CO-AGGREGATION,BACTERIAL-INTESTINAL EPITHELIAL CELLADHESION,BIOFILM FORMATION AND ANTIMICROBIALRESISTANCE OF BACTEROIDES FRAGILIS bacteroides fragilis anearobic bacteria Bile as detergenet Bile salt hydrolase enzyme bacteroides fragilis NCTC9343
TRYPANOSOMA CRUZI: ULTRA STRUCTUREALSTUDIES OF ADHESION,LYSIS AND BIOFILMFORMATION BY SERRATIA MARCESCENS trypanosoma cruzi cused chagas disease Serratia SM365 Attach by D_MANOSE Serratia DB11
STRUCTURE AND FUNCTION OF ESHERICHIA COLIPROTEIN YMGB:APROTEIN CRITICAL FOR BIOFILMFORMATION AND ACID RESISTANCE YMgB gene in biofilm In richment culture of glocose ,decrease motolity of cellular Resistance to acid
REFERENCES 1-Christian m,toutain(2007) Roles of flagellar stators in biofilm formation by pseudomonas aeruginosa 2-Staffan kjellberg and micheal givskov(2007) the biofilm mode of life :mechanism and adaptations 3-M.oliveira s.f nunes (2007)Time course of biofilm formation by staphylococcus aureus and staphylococcus epidermidis mastitis isolates 4-Daniele p.castro Sergio H.seabra (2007) trypanosoma cruzi: ultra structureal studies of adhesion,lysis and biofilm formation by serratia marcescens
5-Lilian pumbwe christopher A.skilbeck(2007) bile salts enhance bacterial co-aggregation, bacterial-intestinal epithelial cell adhesion,biofilm formation and antimicrobial resistance of bacteroides fragilis 6-Kristina mojica danielle Elsey (2007) Quantitative analysis of biofilm EPS uronic acid content 7-A.P.FONSECA J.C SOUSA(2007) effect of shear stress on growth ,adhesion and biofilm formation of pseudomonas aeroginosa with antibiotic_induced morphlogical changes 8-JINTAE LEE Rebecca pag (2007) 9-Structure and function of esherichia coli protein YmgB:Aprotein critical for biofilm formation and acid resistance