1. IN THE NAME OF GOD

2. BIOFILM FORMATION IN
PATHOGENS BACTERIA
Profosser: Dr.ahmad ali PORBABAEI
by: ABBAS MOROVVATI

3. 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

4. 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

5. BIOFILM
 Biofilms consist of:
1-microbes
2-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

6. HISTORY
 Van lion Hook
 zobel
 1913 Sohngen
 1970 enviromental
places
 1971 Marshal
 1973 charakis
 1976 koshreton

7. BIOFILM INFORMATION

8. 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.

9. 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

10. . 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

11. BIOFILM IMAGE BY ESEM

12. 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

13. 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

14. 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

15. 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-

16. 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

17. ROLES FOR FLAGELLAR STATORS IN BIOFILM
FORMATION 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

18. 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

19. 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.

20. 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.

21. GENES / GENE PRODUCTS
Biofilm 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-
Phosphate
Control gene
 16S RNA, not involved in biofilm formation, to be
used as a control

22. TIME COURSE OF BIOFILM FORMATION BY
STAPHYLOCOCCUS AUREUS AND STAPHYLOCOCCUS
EPIDERMIDIS 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

23. EFFECT OF SHEAR STRESS ON GROWTH ,ADHESION AND BIOFILM
FORMATION OF PSEUDOMONAS AEROGINOSA WITH
ANTIBIOTIC_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)

24. BILE SALTS ENHANCE BACTERIAL CO-AGGREGATION,
BACTERIAL-INTESTINAL EPITHELIAL CELL
ADHESION,BIOFILM FORMATION AND ANTIMICROBIAL
RESISTANCE OF BACTEROIDES FRAGILIS
 bacteroides fragilis
anearobic bacteria
 Bile as detergenet
 Bile salt hydrolase
enzyme
 bacteroides fragilis
NCTC9343

25. TRYPANOSOMA CRUZI: ULTRA STRUCTUREAL
STUDIES OF ADHESION,LYSIS AND BIOFILM
FORMATION BY SERRATIA MARCESCENS
 trypanosoma cruzi
cused chagas disease
 Serratia SM365
 Attach by D_MANOSE
 Serratia DB11

26. STRUCTURE AND FUNCTION OF ESHERICHIA COLI
PROTEIN YMGB:APROTEIN CRITICAL FOR BIOFILM
FORMATION AND ACID RESISTANCE
 YMgB gene in biofilm
 In richment culture of
glocose ,decrease
motolity of cellular
 Resistance to acid

27. 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

28.  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