INSTEAD OF INTRODUCTION

BIOFILM TECHNOLOGIES – BIOFILM TECHNOLOGIES – COMMON TERRITORY COMMON TERRITORY

OF BIOTECHNOLOGY AND OF BIOTECHNOLOGY AND NANOTECHNOLOGYNANOTECHNOLOGY

Ludmil Nikolov Ludmil Nikolov Biological FacultyBiological Faculty

of Sofia University of Sofia University “St. Kl. Ohridski”“St. Kl. Ohridski”

INSTEAD OF INSTEAD OF INTRODUCTIONINTRODUCTION

Activity 4.1Activity 4.1.. Nanosciences and Nanotechnologies Nanosciences and Nanotechnologies

4.1.1 Nanosciences and converging sciences:4.1.1 Nanosciences and converging sciences:

4.1.2 Nanotechnologies and converging 4.1.2 Nanotechnologies and converging technologiestechnologies

4.1.3 Health, Safety and Environmental 4.1.3 Health, Safety and Environmental ImpactsImpacts

4.1.24.1.2.. Nanotechnologies and Nanotechnologies and cconverging onverging technologiestechnologies

Pilot lines to study, develop and up-scale Pilot lines to study, develop and up-scale nanotechnology-based processes from nanotechnology-based processes from laboratorylaboratory

Equipment and methods for nanotechnologyEquipment and methods for nanotechnology Analysis of the ethical, regulatory, social and Analysis of the ethical, regulatory, social and

economic environment of nanomedicineeconomic environment of nanomedicine Coordination in nanometrologyCoordination in nanometrology Examining capacity building in Examining capacity building in

nnanobiotechnologyanobiotechnology

The biofilmsThe biofilms

They are well spread in the nature self They are well spread in the nature self organizing formations consisted of:organizing formations consisted of:

Spontaneously fixed cells of: microorganisms, plants or animals on carriers

Produced by themselves substances (EPS - exopolysaccharides, DNA, precipitated salts, etc)

Inclusions of nano- and micro-particles in their structures

Circulating free cells in their structure

BIOFILMS IMPACTSBIOFILMS IMPACTS Biofilms in ecosystemsBiofilms in ecosystems positive rolepositive role in acceleration of natural cycles of biogenic elements - C, N2,

P, S, O2 Biofilms in higher organisms – plants, animals and humans Biofilms in higher organisms – plants, animals and humans controversial roles: negativenegative – dental plaques, diseases prolongation, positivepositive – acceleration of digestion processes in high organisms. Biofilms in artificial systems:Biofilms in artificial systems: negativenegative rolerole – metal protection – biocorrosion, decreasing of the working

characteristics of equipment and apparatus: water transport – decrease of the transport equipment speed, heat exchangers - decrease of heat transfer coefficients, artificial organs, nano-membranes – plug formations.

positive rolepositive role – biofilm reactors, metal leaching acceleration.

BIOFILM TECHNOLOGIESBIOFILM TECHNOLOGIES1. PROCESSING TECHNOLOGIES1. PROCESSING TECHNOLOGIES BIOFILM AS BIOAGENTS (BIOCATALISTS) IN BIOFILM REACTORS IN:BIOFILM AS BIOAGENTS (BIOCATALISTS) IN BIOFILM REACTORS IN:

ENVIRONMENTAL PROTECTION (WASTEWATER TREATMENT, TAIL GAS PURIFICATION)

POTABLE WATER CONDITIONING (BIODENITRIFICATION), BIOMETALLURGY – METAL LEACHING BIOACTIVE SUBSTANCES PRODUCTION ETC.

2. 2. PROTECTING TECHNOLOGIESPROTECTING TECHNOLOGIES (ALSO:“PRESERVING”,“UMPERMISSIBLE”) (ALSO:“PRESERVING”,“UMPERMISSIBLE”)

TO AVOID THE BIOFILM FORMATION (BIOCORROSION OF METALS, DENTAL PLAQUES)

TO DESTROY BIOFILMS IN ARTIFICIAL SYSTEMS (LIKE IN POTABLE WATER SUPLYING SYSTEMS

IN HEALTH CARE – ARTIFICIAL ORGANS WITHOUT BIOFIILMS WATER TRANSPORT EQUIPMENT PROTECTION, NANO-MEMBRANES

WITHOUT BIOFILMS ETC).

THE AIMSTHE AIMS

To draw attention to the analogies of mechanisms of To draw attention to the analogies of mechanisms of biofilms formation and development with some biofilms formation and development with some subjects of nanotechnology.subjects of nanotechnology.

To show similarities of their exploitation orTo show similarities of their exploitation or prevention of development as well as of research prevention of development as well as of research methodology with some nanotechnology.methodology with some nanotechnology.

To illustrate one of the positive rolls, which the To illustrate one of the positive rolls, which the biofilms can play in industry for biofilms can play in industry for highhigh performance performance biofilm reactorsbiofilm reactors design and development design and development

To provoke discussion about the possibility of To provoke discussion about the possibility of transfer transfer of knowledgeof knowledge from one to another scientific fields.from one to another scientific fields.

To give examples of so called “converging To give examples of so called “converging technologies” with nanotechnologies. technologies” with nanotechnologies.

METODOLOGYMETODOLOGYComparative analysisComparative analysisSIMILARITIES AND DIFFERENCES SIMILARITIES AND DIFFERENCES OF BIOFILM TECHNOLOGIES (BIOTECHNOLOGY)OF BIOFILM TECHNOLOGIES (BIOTECHNOLOGY)WITH NANOTECHNOLOGIES (NANOSCIENCE)WITH NANOTECHNOLOGIES (NANOSCIENCE)

The similaritiesThe similarities

They are coming into being due to amalgamation of physical, chemical, physico-chemical (and biological phenomena), accepted as summarizing notion of “spontaneous fixation”.

Mechanisms of biofilm formation are similar to those of thin layers formation in nanotechnologies but in addition – they have all the pluses and minuses of their biological character due to the cells, which are both participants and reason of the formation of these specific living systems.

Diffusion character of structure of biofilm in the terms of system approach.

Some subsystems of biofilm large systems are consisted of macromolecules with nano characteristic parameters – exoploysaccharides, enzymes, nonstoichiomentric compounds, nucleic acids (DNA, RNA etc), parts of cells (organelles, protoplasts etc).

METODOLOGYMETODOLOGYComparative analysisComparative analysis SIMILARITIESSIMILARITIES ANDAND DIFFERENCES DIFFERENCES OF BIOFILM TECHNOLOGIES (BIOTECHNOLOGY)OF BIOFILM TECHNOLOGIES (BIOTECHNOLOGY)WITH NANOTECHNOLOGIES (NANOSCIENCE)WITH NANOTECHNOLOGIES (NANOSCIENCE)

The differencesThe differences

The sizes of the participants of biofilms – microbial cells and moreover - cells of higher organisms (plants or animals) are bigger than the characteristic parameters of subjects of nanotechnologies

Self-organizing character of biofilms as large system Complicated self-created structures of biofilms Living matter in processing systems Necessity of intervene into the processes of self

organization

PECULIARITIES OF BIOFILMS PECULIARITIES OF BIOFILMS AS SELF-ORGANIZING LIVIG SYSTEMSAS SELF-ORGANIZING LIVIG SYSTEMS

1. BIOFILM STRUCTURES 1. BIOFILM STRUCTURES

Fig. 1a. Fig. 1a. Stylized scheme of biofilm system formed on spherical carrier

A – carrier B – biofilm C – fluid 1 – clusters 2 – exopolysaccharides 3 – pores 4 – closed area 5 – cells 6 – microparticles 7 – inert macroparticles 8 – stream lines

a)

1B

3

2

4 5б

5в

5д

6а

7б

6б

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5а

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в

1. BIOFILM STRUCTURES 1. BIOFILM STRUCTURES

Fig. 1b. Fig. 1b. Stylized scheme of biofilm system formed on plane surface carrier

1. Carrier2. Biofilm3. Strongly linked cells in the

biofilm4. Weakly linked cells in the biofilm5. Dead cells6. Stream lines7. Substrate8. Suspended cells9. Pores10. Closed area11. Exsopolysaccharides12. Jarosite.

1 2

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1. BIOFILM STRUCTURES1. BIOFILM STRUCTURES

Fig. 2. Fig. 2. Formation of biofilm structures on the frontier with liquid phase at different scales of liquid flow linear velocity

АА - low velocities ; - low velocities ;

B – middle velocities ; B – middle velocities ; C – high velocities.C – high velocities.1 – piques on the biofilm 1 – piques on the biofilm surface; surface; 2 – pores (channels); 2 – pores (channels); 3 - streamers; 3 - streamers; 4 - microorganisms; 4 - microorganisms; 5 – liquid flow lines;5 – liquid flow lines;6 – closed empty room.6 – closed empty room.

А B C

1 2 4 5 3 6 3 3

1. BIOFILM STRUCTURES1. BIOFILM STRUCTURES

Fig. 3. Fig. 3. Various states of fixed cells at the beginning of biofilm formation

Structures A and B: A and B: 1 - active cells1 - active cells2 - inactive cells 2 - inactive cells

Structure C:Structure C:3 - weakly linked cells3 - weakly linked cells4 - strongly bounded microbial 4 - strongly bounded microbial

cells cells

А B C

1 1 1

2

1 2

3 4

2. DYNAMICS 2. DYNAMICS OF BIOFILM FORMATION and OF BIOFILM FORMATION and FUNCTIONINGFUNCTIONING

Admissions:Admissions:

1. Biofilm formation and functioning is carried out in 1. Biofilm formation and functioning is carried out in open systemsopen systems

2. No mass- and heat transport limitations in the liquid 2. No mass- and heat transport limitations in the liquid phasephase

3. No limitation regarding the constant cells 3. No limitation regarding the constant cells concentrations in the liquid phase concentrations in the liquid phase

Stages of biofilm system developmentStages of biofilm system development

Stage 1Stage 1

Fig. 4Fig. 4аа.. Stage 1.Stage 1. Adaptation of cells to the carrier surface СС – carrier; S – substrate; – carrier; S – substrate; ll – distance from the surface of the carrier; – distance from the surface of the carrier; 1- swimming cells (suspended cell culture); 2- adhered cell1- swimming cells (suspended cell culture); 2- adhered cell

Start of the stage: Start of the stage: cell adaptationcell adaptation End of the stage: End of the stage: singlesingle attached cellsattached cells

S 0 l

S

0

C 1 2

BIOLOGICAL PHENOMENABIOLOGICAL PHENOMENA

swimming cells:swimming cells: Cell adaptation Cell adaptation Swimming cells reproductionSwimming cells reproduction Bioconversion of substrates by swimming cells to:Bioconversion of substrates by swimming cells to:

- products (BAC – biological active compounds) according the aim of - products (BAC – biological active compounds) according the aim of the technologythe technology- EPS on cell surface need for their future fixation- EPS on cell surface need for their future fixation- specific compounds for carrier surface preparation for cell fixation - specific compounds for carrier surface preparation for cell fixation (adsorption) (adsorption) - byproducts- byproducts

fixed cells:fixed cells: only sporadic single attached cellsonly sporadic single attached cells

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA Adsorption of nutrients from the liquid phase Adsorption of specific compounds excreted by cells Adsorption of cells on the carrier surface (spontaneous

fixation) (Cell fixation on base of electrostatic interactions)

RESULTSRESULTS Carrier surface prepared for cell fixation Carrier surface prepared for cell fixation First adhered cells First adhered cells Needed productsNeeded products Prepared surface of cell surface for fixation (adsorption)Prepared surface of cell surface for fixation (adsorption)

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – homogeneous mechanism of – homogeneous mechanism of bioconversion (HMBC)bioconversion (HMBC)

No biofilm formation, isolated cases of attached cellsNo biofilm formation, isolated cases of attached cells Domination of swimming cells productivity Domination of swimming cells productivity

NECESSITY OF INTERVENTION INTO SELF NECESSITY OF INTERVENTION INTO SELF ORGANIZATIONORGANIZATION

In the case of processing technologies - artificial carrier surface In the case of processing technologies - artificial carrier surface modification for acceleration of cell fixation modification for acceleration of cell fixation

In the case of protective (“impermissible”) technologies – In the case of protective (“impermissible”) technologies – deliberated carrier surface treatment to avoid biofilm formation deliberated carrier surface treatment to avoid biofilm formation


Stage 2Stage 2

Formation of cell monolayerFormation of cell monolayer

Fig.4b. Fig.4b. Stage 2. Stage 2. Formation of cell monolayer

3 - adhered cells enveloped by exopolysacharides 3 - adhered cells enveloped by exopolysacharides

Start of the stage:Start of the stage: single single attached cells;attached cells; End of the stage:End of the stage: monocell layer. monocell layer.

l

S

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swimming cells:swimming cells: Cell adaptationCell adaptation Swimming cells reproductionSwimming cells reproduction Bioconversion of substrates by swimming cells to:Bioconversion of substrates by swimming cells to:

- products (BAC – biological active compounds) according the aim of the technology - products (BAC – biological active compounds) according the aim of the technology - substances for biofilm formation and its structuring (EPS, precipitated salts, etc) - substances for biofilm formation and its structuring (EPS, precipitated salts, etc) - byproducts - byproducts

fixed cells:fixed cells: Beginning of the fixed cells reproduction on the carrier surface – isolated casesBeginning of the fixed cells reproduction on the carrier surface – isolated cases Bioconversion of substrates by fixed cells to:Bioconversion of substrates by fixed cells to:

- products (BAC – biological active compounds) according the aim of the technology - products (BAC – biological active compounds) according the aim of the technology - substances for biofilm formation and its structuring (EPS, precipitated salts, etc) - substances for biofilm formation and its structuring (EPS, precipitated salts, etc) - byproducts - byproducts

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA

Adsorption of nutrients from the liquid phase Adsorption of nutrients from the liquid phase Adsorption of specific compounds excreted by cells Adsorption of specific compounds excreted by cells Adsorption of cells on the carrier surface (spontaneous fixation)Adsorption of cells on the carrier surface (spontaneous fixation) (Cell fixation on base of electrostatic interactions(Cell fixation on base of electrostatic interactions) )

RESULTS:RESULTS: Envelopment of cells with substances for biofilm formation Envelopment of cells with substances for biofilm formation

mainly EPS.mainly EPS. First cell configurations on the carrier (cell monolayer First cell configurations on the carrier (cell monolayer

formation)formation) Fixed cell reproductionFixed cell reproduction Needed productsNeeded products

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – homogeneous-heterogeneous – homogeneous-heterogeneous mechanism (HHMBC)mechanism (HHMBC)

Domination of swimming cells productivity Domination of swimming cells productivity Fixed cells start to participate in bioconversionFixed cells start to participate in bioconversion

INTERVENTION INTO SELF ORGANIZATIONINTERVENTION INTO SELF ORGANIZATION In processing technologies - no needIn processing technologies - no need In protective (“impermissible”) technologies – mechanical In protective (“impermissible”) technologies – mechanical

means: high shear stress around carrier surface, scraping off means: high shear stress around carrier surface, scraping off biofilm formed, biocides introduction in the liquid phase etc. biofilm formed, biocides introduction in the liquid phase etc.


Stage 3Stage 3

Biofilm structure formationBiofilm structure formation

Fig. 4 c. Fig. 4 c. Stage 3. Stage 3. Formation of the biofilm structure. First critical thickness of the biofilm - δ1Cr

Start of the stage:Start of the stage: monocell layer; monocell layer; End of the stage:End of the stage: polycell layer, first critical biofilm thickness -δ polycell layer, first critical biofilm thickness -δ1Cr1Cr..

S l

S

0

C

δ1Cr 0


swimming cells:swimming cells: Swimming cells reproductionSwimming cells reproduction Bioconversion of substrates by swimming cells to:Bioconversion of substrates by swimming cells to:

- products (BAC – biological active compounds) according the aim of the - products (BAC – biological active compounds) according the aim of the technologytechnology

- substances for biofilm formation and its structuring (EPS, precipitated - substances for biofilm formation and its structuring (EPS, precipitated salts, etc)salts, etc)- byproducts- byproducts

fixed cells:fixed cells: Fixed cells reproduction on the biofilm surface and in the biofilm volumeFixed cells reproduction on the biofilm surface and in the biofilm volume Bioconversion of substrates by fixed cells to:Bioconversion of substrates by fixed cells to:

- products (BAC – biological active compounds) according the aim of the - products (BAC – biological active compounds) according the aim of the technologytechnology- substances for biofilm formation and its structuring (EPS, precipitated - substances for biofilm formation and its structuring (EPS, precipitated salts, etc)salts, etc)- byproducts- byproducts

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA Adsorption of cells on the biofilm surface (spontaneous fixation)Adsorption of cells on the biofilm surface (spontaneous fixation)

RESULTS:RESULTS: First cell configurations on the carrier (cell monolayer formation)First cell configurations on the carrier (cell monolayer formation) Fixed cell reproductionFixed cell reproduction Biofilm structure formationBiofilm structure formation Needed productsNeeded products

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – mixed homogeneous-heterogeneous – mixed homogeneous-heterogeneous mechanism mechanism

Domination of fixed cells productivity Domination of fixed cells productivity Bioconversion without internal diffusion limitations Bioconversion without internal diffusion limitations

Conditions for pure biofilm kinetics investigationConditions for pure biofilm kinetics investigation

INTERVENTION INTO SELF ORGANIZATIONINTERVENTION INTO SELF ORGANIZATION In processing technologies In processing technologies - artificial biofilm thickness control at the - artificial biofilm thickness control at the

first critical value - δ1first critical value - δ1Cr Cr (by the biofilm reactor design)(by the biofilm reactor design) In protective (‘impermissible”) technologiesIn protective (‘impermissible”) technologies – mechanical means: high – mechanical means: high

shear stress around carrier surface, scraping off biofilm formed, shear stress around carrier surface, scraping off biofilm formed, chemical means: biocides introduction in the liquid phase etc. chemical means: biocides introduction in the liquid phase etc.


Stage 4Stage 4

Stable growth of biofilm systemStable growth of biofilm system

Fig. 4 d. Fig. 4 d. Stage 4. Stage 4. Stable biofilm growth. Second critical biofilm thickness - δ2Cr

Start of the stage:Start of the stage: first critical biofilm thickness - first critical biofilm thickness -- - δδ11CrCr

End of the stage:End of the stage: second critical biofilm thickness - second critical biofilm thickness -- - δδ22CrCr

l δ2Cr

S

C S

0 0

BIOLOGICAL PHENOMENABIOLOGICAL PHENOMENAswimming cells:swimming cells: Swimming cells reproductionSwimming cells reproduction Bioconversion of substrates by swimming cells to:Bioconversion of substrates by swimming cells to:

- products (BAC – biological active compounds) according the aim of the - products (BAC – biological active compounds) according the aim of the technologytechnology- substances for biofilm formation and its structuring - EPS on cell surface- substances for biofilm formation and its structuring - EPS on cell surface- byproducts- byproducts

fixed cells: fixed cells: Fixed cells reproduction in the biofilm surface and volumeFixed cells reproduction in the biofilm surface and volume Bioconversion of substrates by fixed cells to:Bioconversion of substrates by fixed cells to:

- products (BAC – biological active compounds) according the aim of the - products (BAC – biological active compounds) according the aim of the technologytechnology- substances for biofilm formation and its structuring (EPS, precipitated salts, - substances for biofilm formation and its structuring (EPS, precipitated salts, etc)etc)- byproducts- byproducts- new products obtained due to change of metabolism with substrate limited - new products obtained due to change of metabolism with substrate limited kineticskinetics

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA Adsorption of cells on the biofilm surfaceAdsorption of cells on the biofilm surface Internal diffusion limitation in the zone behind δInternal diffusion limitation in the zone behind δ11CrCr of biofilm of biofilm

RESULTS:RESULTS: Well developed biofilm Well developed biofilm Fixed cell reproduction in the biofilm volume and surfaceFixed cell reproduction in the biofilm volume and surface Needed products according the technology aimNeeded products according the technology aim New products (usually undesired, but sometimes quite possible to be useful)New products (usually undesired, but sometimes quite possible to be useful) Linear dependence of biofilm thickness development on timeLinear dependence of biofilm thickness development on time Second critical biofilm thickness - δSecond critical biofilm thickness - δ22CrCr – S = 0 at the carrier surface – S = 0 at the carrier surface

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – – mixed homogeneous-heterogeneous mechanism (HHM)mixed homogeneous-heterogeneous mechanism (HHM) Domination of fixed cells productivity Domination of fixed cells productivity Bioconversion under internal diffusion limitationsBioconversion under internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations New products due to bioconversion under changed condition in the biofilmNew products due to bioconversion under changed condition in the biofilm Biofilm system is still stable Biofilm system is still stable

INTERVENTION INTO SELF ORGANIZATIONINTERVENTION INTO SELF ORGANIZATION In processing technologiesIn processing technologies - biofilm thickness control at desired values - higher than δ - biofilm thickness control at desired values - higher than δ11CrCr (if (if

necessary)necessary) In protective (“impermissing)In protective (“impermissing) technologies - mechanical means: high shear stress around technologies - mechanical means: high shear stress around

carrier surface, scraping off biofilm formed, chemical means: biocides introduction in thecarrier surface, scraping off biofilm formed, chemical means: biocides introduction in the liquid phase etc. liquid phase etc.


Stage 5Stage 5

Uncontrolled and unstable biofilm growthUncontrolled and unstable biofilm growth

Fig.4 e. Fig.4 e. Stage 5. Stage 5. Uncontrolled and unstable biofilm growth. Cavities formation

4 – cavity4 – cavity

Start of the stage:Start of the stage: second critical biofilm thickness -- δ second critical biofilm thickness -- δ2Cr2Cr End of the stage:End of the stage: cavities formation cavities formation

C S

S

l

0 0

4



- products (BAC – biological active compounds) according the aim of the technology- products (BAC – biological active compounds) according the aim of the technology- substances for biofilm formation and its structuring (EPS, precipitated salts, etc)- substances for biofilm formation and its structuring (EPS, precipitated salts, etc)- byproducts- byproducts


- products (BAC – biological active compounds) according the aim of the technology- products (BAC – biological active compounds) according the aim of the technology- substances for biofilm formation and its structuring (EPS, precipitated salts, etc)- substances for biofilm formation and its structuring (EPS, precipitated salts, etc)- byproducts- byproducts- new products obtained due to change of metabolism with substrate limited kinetics- new products obtained due to change of metabolism with substrate limited kinetics

Cell decay and lysis nearby the carrier surface due to lack of substratesCell decay and lysis nearby the carrier surface due to lack of substrates Products of the decay and lysis in the biofilm volume (possibly also in the liquid) Products of the decay and lysis in the biofilm volume (possibly also in the liquid) Cavities formationCavities formation

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA Adsorption of cells on the biofilm surfaceAdsorption of cells on the biofilm surface Diffusion limitation in the biofilm Diffusion limitation in the biofilm Detachment of parts of biofilm from carrier surface due to cells decay and lysisDetachment of parts of biofilm from carrier surface due to cells decay and lysis

RESULTS:RESULTS: Thick biofilm with cavitiesThick biofilm with cavities Fixed cell reproduction in the biofilm volume and surfaceFixed cell reproduction in the biofilm volume and surface Needed products according to technology aimNeeded products according to technology aim New products (usually undesired, but sometimes quite possible to be useful) New products (usually undesired, but sometimes quite possible to be useful) Non linear dependence of biofilm thickness development on timeNon linear dependence of biofilm thickness development on time Lack of substrates in the deeper biofilm layersLack of substrates in the deeper biofilm layers Products of cells decay and lysisProducts of cells decay and lysis

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – – beginning of destruction, HHMbeginning of destruction, HHM Domination of fixed cells productivity Domination of fixed cells productivity Bioconversion under internal diffusion limitationsBioconversion under internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Production of new products due to changed condition in the biofilmProduction of new products due to changed condition in the biofilm Biofilm system is in the state of the beginning of its destructionBiofilm system is in the state of the beginning of its destruction

INTERVENTION INTO SELF ORGANIZATIONINTERVENTION INTO SELF ORGANIZATION In processing technologiesIn processing technologies - no need - biofilm system destruction is starting and - no need - biofilm system destruction is starting and

this process is uncontrollablethis process is uncontrollable In protective (“impermissible”) technologiesIn protective (“impermissible”) technologies – mechanical means: high shear – mechanical means: high shear

stress around carrier surface, scraping off biofilm formed, chemical means: stress around carrier surface, scraping off biofilm formed, chemical means: biocides introduction in the liquid phase etc. biocides introduction in the liquid phase etc.


Stage 6Stage 6

Biofilm destructionBiofilm destruction

Fig. 4 f. Fig. 4 f. Stage 6. Stage 6. Biofilm destruction. Third critical biofilm thickness- δ3Cr

5 – detached part of the biofilm structure5 – detached part of the biofilm structure

Start of the stage:Start of the stage: cavities formation cavities formation End of the stage:End of the stage: detachment of parts of biofilm volumedetachment of parts of biofilm volume

C S

S

δ3Cr l 0

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5



- products (BAC – biological active compounds) according the aim of the technology- products (BAC – biological active compounds) according the aim of the technology- substances for biofilm formation and its structuring - EPS on the cell surface- substances for biofilm formation and its structuring - EPS on the cell surface- byproducts.- byproducts.



Cell decay and lysis nearby the carrier surfaceCell decay and lysis nearby the carrier surface Products of the decay and lysis in the biofilm volume (possibly also in the liquid) Products of the decay and lysis in the biofilm volume (possibly also in the liquid)

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA Adsorption of cells on the biofilm surfaceAdsorption of cells on the biofilm surface Diffusion limitation in the biofilm volumeDiffusion limitation in the biofilm volume Detachment of cells nearby the carrier surface due to cells decay and lysisDetachment of cells nearby the carrier surface due to cells decay and lysis Detachment of biofilm parts from the outer layers to the liquid phaseDetachment of biofilm parts from the outer layers to the liquid phase

RESULTS:RESULTS: Destroyed biofilmDestroyed biofilm Fixed cell reproduction in the biofilm volume and surfaceFixed cell reproduction in the biofilm volume and surface Needed products according to the technology aimNeeded products according to the technology aim New products (usually undesired, but sometimes quite possible to be useful) New products (usually undesired, but sometimes quite possible to be useful) Non linear dependence of biofilm thickness on time, decrease of biofilm volumeNon linear dependence of biofilm thickness on time, decrease of biofilm volume Lack of substrates in the deeper biofilm layersLack of substrates in the deeper biofilm layers Cavities formationCavities formation Products of cells decay and lysisProducts of cells decay and lysis Detached biofilm parts swimming in the liquid phaseDetached biofilm parts swimming in the liquid phase Third critical biofilm thickness – δThird critical biofilm thickness – δ33CrCr

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – – system destruction; HHMsystem destruction; HHM Domination of fixed cells productivity :Domination of fixed cells productivity :

- in the biofilmin the biofilmin the detached biofilm parts swimming in the liquid phase - in the detached biofilm parts swimming in the liquid phase

Bioconversion under internal diffusion limitationsBioconversion under internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Production of new products due to changed conditionProduction of new products due to changed condition in the biofilmin the biofilm Biofilm system is in destructionBiofilm system is in destruction

INTERVENTION INTO SELF ORGANIZATIONINTERVENTION INTO SELF ORGANIZATION In processing technologies - In processing technologies - artificial retention of detached biofilm parts in the liquid phase artificial retention of detached biofilm parts in the liquid phase

(if necessary), i.e. in the bioconversion zone.(if necessary), i.e. in the bioconversion zone. In protective (“impermissible”) technologiesIn protective (“impermissible”) technologies – mechanical means: high shear stress around – mechanical means: high shear stress around

carrier surface, scraping off biofilm formed, chemical means: biocides introduction in the carrier surface, scraping off biofilm formed, chemical means: biocides introduction in the liquid phase etc.liquid phase etc.


Stage 7Stage 7

Restart of new biofilm formation. Simultaneous realization of all the stages.Restart of new biofilm formation. Simultaneous realization of all the stages.

Fig. 4 g. Fig. 4 g. Stage 7. Stage 7. Restart of new biofilm formation

Start of the stage:Start of the stage: cavities formation cavities formation End of the stage:End of the stage: detachment of parts of biofilm volumedetachment of parts of biofilm volume

C S

l 0

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5

1 2

3

4

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S

1 - swimming cells (suspended cell culture)1 - swimming cells (suspended cell culture)2- adhered cell2- adhered cell3 - adhered cells enveloped by exopolysacharides 3 - adhered cells enveloped by exopolysacharides 4 - cavity4 - cavity5 - detached part of the biofilm structure5 - detached part of the biofilm structure6 - new attached cells 6 - new attached cells



- products (BAC – biological active compounds) according the aim of the technology- products (BAC – biological active compounds) according the aim of the technology- substances for biofilm formation and its structuring - EPS on the cell surface- substances for biofilm formation and its structuring - EPS on the cell surface- byproducts- byproducts



Cell decay and lysis nearby the carrier surfaceCell decay and lysis nearby the carrier surface Products of the decay and lysis in the biofilm volume (possibly also in the liquid) Products of the decay and lysis in the biofilm volume (possibly also in the liquid)

PHYSICO-CHEMICAL PHENOMENAPHYSICO-CHEMICAL PHENOMENA Adsorption of cells on the biofilm surfaceAdsorption of cells on the biofilm surface Diffusion limitation in the biofilm volumeDiffusion limitation in the biofilm volume Detachment of cells nearby the carrier surface due to cells decay and lysisDetachment of cells nearby the carrier surface due to cells decay and lysis Detachment of biofilm parts from the outer layers to the liquid phaseDetachment of biofilm parts from the outer layers to the liquid phase

RESULTS:RESULTS: Destroyed biofilmDestroyed biofilm Fixed cell reproduction in the biofilm volume and surfaceFixed cell reproduction in the biofilm volume and surface Needed products according to the technology aimNeeded products according to the technology aim New products (usually undesired, but sometimes quite possible to be useful) New products (usually undesired, but sometimes quite possible to be useful) Non linear dependence of biofilm thickness on time, decrease of biofilm volumeNon linear dependence of biofilm thickness on time, decrease of biofilm volume Lack of substrates in the deeper biofilm layersLack of substrates in the deeper biofilm layers Cavities formationCavities formation Products of cells decay and lysisProducts of cells decay and lysis Detached biofilm parts swimming in the liquid phaseDetached biofilm parts swimming in the liquid phase Restart of biofilm formation Restart of biofilm formation All the phases of biofilm dynamics are performing simultaneouslyAll the phases of biofilm dynamics are performing simultaneously Repeated biofilm growth:Repeated biofilm growth:

- on the carrier using places of detached biofilm parts- on the carrier using places of detached biofilm parts- on the outer biofilm surface- on the outer biofilm surface

BIOFILM SYSTEM STATEBIOFILM SYSTEM STATE – system renovation, HHM, all the stage existence – system renovation, HHM, all the stage existence Domination of fixed cells productivity:Domination of fixed cells productivity:

- in the biofilmin the biofilmin the detached biofilm parts swimming in the liquid phase - in the detached biofilm parts swimming in the liquid phase

Bioconversion under internal diffusion limitationsBioconversion under internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Conditions for biofilm kinetics investigation with internal diffusion limitations Production of new products due to changed conditionProduction of new products due to changed condition in the biofilmin the biofilm Biofilm system is in renovationBiofilm system is in renovation

INTERVENTION INTO SELF ORGANIZATIONINTERVENTION INTO SELF ORGANIZATION In processing technologies - In processing technologies - artificial retention of detached biofilm parts in the liquid phase (if artificial retention of detached biofilm parts in the liquid phase (if

necessary)necessary) In protective (“impermissible”) technologiesIn protective (“impermissible”) technologies – mechanical means: high shear stress around carrier – mechanical means: high shear stress around carrier

surface, scraping off biofilm formed, chemical means: biocides introduction in the liquid phase etc.surface, scraping off biofilm formed, chemical means: biocides introduction in the liquid phase etc.

APPLICATIONS inAPPLICATIONS inBIOFILM REACTOR DESIGN AND DEVELOPMENTBIOFILM REACTOR DESIGN AND DEVELOPMENT HIGH PERFORMANCE INVERSE FLUIDIZED BED BIOFILM HIGH PERFORMANCE INVERSE FLUIDIZED BED BIOFILM REACTORSREACTORS

P

Air

bioparticle

support

biofilm

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Stage 1

Stage 2

Stage 3

APPLICATIONS inAPPLICATIONS inBIOFILM REACTOR DESIGN AND DEVELOPMENTBIOFILM REACTOR DESIGN AND DEVELOPMENT HIGH PERFORMANCE INVERSE FLUIDIZED BED BIOFILM HIGH PERFORMANCE INVERSE FLUIDIZED BED BIOFILM REACTORSREACTORS

b)

S

P

Air

1

2

3

4

5

6

S 0 l

S

0

C 1 2

l

S

0

S C

0

3

S l

S

0

C

δ1Cr 0

Stage 1

Stage 2

Stage 3

APPLICATIONS inAPPLICATIONS inBIOFILM REACTOR DESIGN AND DEVELOPMENTBIOFILM REACTOR DESIGN AND DEVELOPMENT

HIGH PERFORMANCE BIODISK REACTORHIGH PERFORMANCE BIODISK REACTOR

inlet

outlet for sediments

biofilm

disk

outlet

wire-net

fragments from biofilm

C S

l 0

0

5

1 2

3

4

6

S Stage 7

CONCLUSIONSCONCLUSIONS

COMON TERITORYCOMON TERITORYOF BIOTECHNOLOGY AND NANOTECHNOLOGY OF BIOTECHNOLOGY AND NANOTECHNOLOGY IS IN ACTIVE INTERVENTION IN THE DINAMICS OFIS IN ACTIVE INTERVENTION IN THE DINAMICS OFBIOFILM SELFORGANIZATION AS FOLLOW:BIOFILM SELFORGANIZATION AS FOLLOW:

1. 1. PROCESSING TECHNOLOGIESPROCESSING TECHNOLOGIES To accelerate the first stage of biofilm formation by means of To accelerate the first stage of biofilm formation by means of

introduction of specific compounds for modification of the introduction of specific compounds for modification of the carrier surfacecarrier surface

2. 2. PROTECTING TECHNOLOGIESPROTECTING TECHNOLOGIES To find new appropriate approaches to eliminate To find new appropriate approaches to eliminate any possibility any possibility

of biofilm formation of biofilm formation on the material surfaceson the material surfaces To develop new nanotechnologies for biofilm-defended To develop new nanotechnologies for biofilm-defended

membranes production for ultra filtration under industrial membranes production for ultra filtration under industrial conditionsconditions

Documents

INSTEAD OF INTRODUCTION