bacteria and biocide

BACTERIA AND BIOCIDES

CLASSIFICATION BASED ON RESPONSE TO OXYGEN AEROBIC ANAEROBIC FACULTATIVE MICROAEROPHILIC EH REQUIREMENTS

CLASSIFICATION ACCORDING TO ENVIRONMENTAL PREFERENCE

BARAPHILES THERMOPILES MESOPHILES PSYCHROPHILES HALOPHILES ACIDOPHILES NEUTROPHILES ETC. I.E., BAROPHILLIC THERMOPHILES

[SESSILE VS PLANKTONIC]

TYPES OF BACTERIA THAT ARE IMPORTANT IN THE OIL FIELD Sulfate reducing bacteria (SRBs)

desulfovibrio desulfomaculum

Anaerobic Bacteria Aerobic Bacteria Facultative Bacteria Iron Bacteria Others

sulfur oxidizing nitrate reducing methanogens fungi

[SESSILE VS PLANKTONIC]

SIGNS OF BACTERIAL CONTAMINATION

BLACK WATER [H2S]

[FeS]/SOLIDS PLUGGING / FILTER PROBLEMS /

HIGH INJECTIVITY PRESSURES PITTING CORROSION - PATTERNS HIGH BACTERIA COUNTS

BREEDING SITES

SLIME FORMERS

These bacteria need oxygen to grow. Although oil field waters are naturally very low in oxygen in many instances air can get into field water through system leaks.

OPEN PONDS SHALLOW WATER SUPPLY WELLS ON PRODUCTION EQUIPMENT ON TUBING

BREEDING SITES

SRB’SCan be found in many areas. Will only grow in an

environment that has low concentrations of oxygen.

RAT HOLES PACKER FLUIDS HEATER TREATERS FILTER BEDS AT OIL/WATER INTERFACES UNDER DEPOSITS

Intake Pumps

Strainer

Filtration

Deaeration

ChemicalTreatment

Injection Pump

Reservoir

ProcessCooling

DesalinationMudMake-up

Dump WaterStore

MudStore

WATER INJECTION SYSTEM

(1)

(2)

(3)

(4)

(5)

(6)

(7) (8)

( ) Sampling Point

Sampling Notes:

1. General organisms for water quality; blooms ; efficacy of chlorination; consolidation techniques for sulfate-reducing bacteria.

2. General organisms; clogging filters; backwash schedules; souring of deep-beds.

3. Sulfate-reducing bacteria.

4. Sulfate-reducing bacteria probably by consolidation techniques; check biocide efficiency.

5. Backflushing of reservoir; check sulfate-reducing bacteria and sulfur.

6. Heat exchangers, etc; general organisms slime formation and loss of heat transfer.

7. Quality of drinking water.

8. Blooms; sulfate-reducing bacteria; odor nuisance of mud in store.

Reservoir

Wellhead

High PressureSeparator

Low PressureSeparator

Pipeline

Storage

Storage

Tanker

Oily Water

(1)

(2A)

(2B)

(3)

(4)

(5)

(6)

PRODUCTION SYSTEM

( ) Sampling Point

Sampling Notes: 1. Sulfate-reducing bacteria levels after shut-ins; sulfate-

reducing bacteria on downloads SV; sulfate-reducing bacteria in sea water breakthrough.

2. Sulfate-reducing bacteria in water bottoms. 3. Sulfate-reducing bacteria in pipeline water at landfalls;

pigging debris; biocide efficacy tests. 4. Sulfate-reducing bacteria in tank bottoms. 5. Sulfate-reducing bacteria in offshore sea water ballasted

crude storage tanks; general oil degrading organisms. 6. General oil degrading organisms; sulfate-reducing bacteria.

Often main source of contamination of pipelines.

ENUMERATION / IDENTIFICATION OF BACTERIA Serial dilution followed by culturing in the appropriate

media Epifluorescence A.T.P Photometry Microscopic techniques “Fast” count methods

A true identification of bacteria is very complicated. This is generally not carried out in an oil field operation.

*sanicheck*rapid check

CULTURING OF BACTERIA

In order to successfully culture bacteria an environment needs to be provided that allows them to grow and multiply.

Requirements:Specificity

[Salt]

pH

Eh-(poising agents)

Temperature of incubation

Visualization

Time to develop growth

Availability of media

MEDIA REQUIREMENTS

SRB ANAEROBES AEROBESFood source lactic acid

acetic acidothers

various sugars protein extracts

Cations Na, Ca, Mg, Fe, etc Na, Ca, Mg, Fe, etc Na, Ca, Mg, Fe, etcSulfate + + + + +pH ~7 ~7 ~7Eh (poising Agent) + + -General Nutrients yeast yeastSalts Adjust to suit

organism

CULTURE MEDIA

KH2PO4 0.5gNH4CL 1.0gCaSO4 1.0gMgSO4

. 7H2O 2.0g

yeast extract 1.0gNa lactate 3.5gNa acetate 2.8gAsorbic acid 0.1gThioglycollic acid 0.1gFeSO4

. 7H2O 0.5g

pH AdjustCaClMgCl2 AdjustNaClH2O Add up to 1000ml

SRB Broth(Modified Postgate B)

CULTURE MEDIA

glucose 10.0gKNO3 0.6gNa2HPO4 0.45gMgSO4

. 7H2O 0.1g

FeSO4 . 7H2O 0.001g

yeast extract 3.0gNH4Cl 0.5gCysteine-HCl 0.5gpH 7.0NaCl adjust

H2O Add up to 1000ml

General Anaerobic Broth

CULTURE MEDIA

Beef Extract 3.0gPeptone 5.0gpH 7.0NaCl adjustH2O Add up to 1000 ml

API Aerobic Broth

DO’S AND DON’TS ON BACTERIA CULTURING

CRITICALLY IMPORTANT Adjust salinity to match field salt level. When in between go

to lower chloride. Incubate - temperature, time Darkness - Do not expose to UV Shoot a number of bottles (6) to define severity of problem Safety - needle destruction, bottle disposal, letter of

justification Observe shelf life Thermo-heat the media to avoid killing thermophiles


MAY BE IMPORTANT Changing syringes Alcohol swabbing Volume of sample-precision 1ml +/- 0.2 ml Gauge of needle - avoid large diameter Media manufacturer - stay with one Postgate v. API for SRB


DOES NOT MATTER Drawing vacuum on bottle before withdrawing

needle Size of syringe

SERIAL DILUTION

Bottles ShowingGrowth

Bact. Count (colonies/ml)

1 1-102 10-1003 100-10004 1000-10,0005 10,000-100,000

If the above procedures are carried out in triplicate (or higher)there are statistical tables available for more accurate counts

SERIAL DILUTION

SAMPLE 0-10 10-100 etc. etc. etc.

PLUGGING PROBLEMS

Bacteria can cause plugging and reduced flow in filters, down hole formations and lines. They can coat the walls of heat exchangers and reduce their efficiency. Formation of insoluble salts (FeS) Formation of slime and cell debris

HOW DO BACTERIA CAUSE CORROSION

-H2S production

SO4-2 + H2 H2S

sulfate hydrogen sulfide

Acid production

sugars various acids(other substrates) (lactic, acetic, propionic)

CO2 + H2

CATHODIC DEPOLARIZATION

2 ELECTRONS

From the anodic metal oxidation

+2H+ 2H H2

In the cell cytochrome systemoxidation of hydrogenreduction of sulfate

SO4-2

H2O + S-2 + Energy

AssimilationReactions

ADP

ATP

BACTERIAL CONTROL WITH BIOCIDES

Lab procedures Compatibility tests (biocide c field fluids)

c other treatment chemicals Time kill tests. This procedure evaluates the amount of chemical

needed and amount of contact time required for a biocide to kill the bacteria or significantly reduce their population density. This test measures the bactericidal potency of a biocide.

Bacteriostatic test. This procedure evaluates the ability of a reagent to stop the growth of (but not necessary kill) bacteria.

Biocide studies on bacterial films. These techniques are used to measure the effect of biocides on sessile bacteria that have been grown on a surface.


[H2S]

Corrosion rates Measurements of injectivity pressure filter plugging, total

solids etc.

Field monitors and surveys (establish base lines)Planktonic Sessile

appropriate samplepoints (bug vials)

Robbins sidestream device

pinpoint specific areasof high conc.

Robbins in-line devicecouponspipe sections etc.


TREATMENT PROCEDURES: Continuous vs slug Specific sites vs general Correlation with lab data Treatment in conjuction with a surfactant Treatment in conjuction with mechanical devices

Butterworth Sprayers

Pigs

Other clean up procedure

AN EXAMPLE OF A TREATMENT REGIMEN

OXYGEN CORROSION

SCAVENGERS INHIBITION PIG BIOCIDE PIG

PIPELINE

BIOCIDES MECHANISMS OF ACTION

Interfere with fundamental cell requirements denaturation of proteins and enzymes disruption of lipids cause cell wall disruption damage / inactivation of DNA, RNA

Most biocides have pluralistic modes of action Significance of resistance

Glycocalyx formation Specificity of action is rare

Synergisms

BIOCIDE REGISTRATION / LABELING

EPA LABEL REQUIREMENTS

BIOCIDES

DESCRIPTION AND AVAILABILITY

BIOCIDES (MISCELLANEOUS)Other biocideImidazolines NapervilleSulfones NapervilleSodium Hypochlorite NapervilleBenzothiazole NapervilleTin oxide NapervilleThiocyanate NapervilleGuanidine (VANTOCIL)

Europe

Copper Sulfate NapervilleMetronidazole Not Available

BIOSURFACANTS (biodispersants)

EC1082A Diamine plus surfactants

EC9047A Surfactant only

EC1088A Diamine plus surfactants

SAMPLING OF BACTERIA / REVIEW

Sessile bacteria - need to obtain the bacteria from the surface.

Planktonic bacteria - need to obtain the bacteria out of the liquid.

The procedures for obtaining sessile counts are somewhat more cumbersome than for planktonic but utilize the same fundamental principles.

Important Factors: Eh requirements oxygen concentration sampling techniques

BIOCIDE INCOMPATIBILITIES

Reactions with other chemicals. Temperature sensitivity. Sensitivity to pH. Solubility in brines. Foaming. Emulsion Formation

BACTERIAL METABOLISM

TYPE

Respiration

Anaerobic respiration

Fermentation

EXAMPLEC6H12O6 + 6O2 6CO2 + energy

CH3COOH + SO4-2 2CO2 + H2S

+2OH- + energy

C6H12O6 2CO2 + 2C2H5OH + energy

CONCLUSION

The determination of what problems exist in a field can be difficult and frustrating.

Many times more than one type of corrosion is taking place simultaneously. In these instances it may be very hard to determine which one (or more) is the significant factor that needs to be controlled.

RELATIONSHIP OF TEST DATA TO FIELD SYSTEMS

BATCH TREATMENT CONTINUOUS TREATMENT

CHEMICALS USED FOR BACTERIAL CONTROL

BACTERICIDES - CHEMICALS WHICH KILL BACTERIA

BACTERIOSTATS - CHEMICALS WHICH RETARD OR INHIBIT THE GROWTH OF BACTERIA

MICROBIOCIDES - CHEMICALS WHICH KILL OTHER FORMS OF LIFE IN ADDITION TO BACTERIA

MICROBIOSTATS - CHEMICALS WHICH RETARD OR INHIBIT THE GROWTH OF OTHER FORMS OF LIFE IN ADDITION TO BACTERIA

SAMPLING / ENUMERATION OF BACTERIA

PROCEDURES WITH PLANKTONIC BACTERIA

PROCEDURE WITH SESSILE BACTERIA

TAXONOMY

Kingdom - Plant

Phylum - Thallophyta

Class - Schizomycetes

Order - Pseudomonadales

Family - Spirillaceae

Genus - Desulfovibrio

Species - ie. vulgaris

desulfuricans

africans

MISC. CLASSIFICATIONS

Spore formers Staining techniques

gram pos

gram neg

gram variable

MORPHOLOGICAL CLASSIFICATIONS

Rods Curved (sigmoid) (like the letter C) Vibrio (like a comma or s shaped) Spiral Semilunar (cresent shaped) Coccoid (round) Filamentous Flageller classifications

peritrichous (uniform over body) monotrichous (one flagella/cell) lophotrichous (dist-about one end)

RememberBacteria can exhibit polymorphism or pleomorphism-especially under conditions of

stress.

CLASSIFICATION OF BACTERIA

There are several ways in which bacteria are grouped.Many of these depend on what particular aspect of the organism is of interest or is being studied.

CLASSIFICATION OF MICROORGANISMS

Algae - Contain Chlorophyll, Require Sunlight

Fungi - Do Not Contain Chlorophyll

*Bacteria - Some Properties Common To Both Algae and Fungi

INTRODUCTION

PRACTICAL THEORETICAL INTEREST HANDOUTS

BIOCIDES

NAME DESCRIPTION AVAILABILITYHouston Europe Naperville

Glutaraldehyde Dialdehyde EC6111A EC9131A yesEC6112A EC9128A

Glut mixtures with quats EC6109A (1) yesEC6110A

Glut mixtures with other chemicals no (1) yes

Formaldehyde no(2) 7-6444 noFormaldehyde mix with other actives no EC9135A no

Diamines R-N-C-C-C-N- EC6106A 7-7672 yesEC6107A EC6210A

Diamine mixtures with other chemicals no (1) yes R+

Quats R-N-R EC6116A EC9123A yes R

Quat mixtures with other chemicals no (1) yes

Isothiazoline Heterocyclic (KATHON) EC6114A no yes S

Thiocarbamates -N-C-S- ASP744 no yesDryocide

THPS Phosphonium quat (3) EC9126A (3)THPS mixtures with other acivites no 7-6450 no

DBNPA Brominated amide EC6116A no yes

Triazine Heterocyclic EC6113A no yes

Dazomet Heterocyclic EC6115A no yes

DIKIOR Chlorine dioxide (4) no no

(1) More than one available(2) Not registered for oilfield use in the USA.(3) Will be added to the product line as soon as it clears the EPA.(4) Will be added to the product line.

SCHEMATIC OF FLUSH MOUNTED BIOPROBE

~2”

PLASTIC FRONT (GLUED ONTO METAL PLATE)

THREADED BACK SIDE

ALLEN SCREW(TO HOLD STUD IN PLACE)

STUD (BIOBULLET)

The flush mounted bioprobe is threaded in the back and screws directly into the line. The plastic on the front isolates the removable studs from the rest of the metal probe and thus eliminates galvanic corrosion. The field water flows over the studs in the line. Bacteria, if present can settle on and stick to the studs. The studs can be removed and the number of bacteria present on the surface is determined.

BUTYL RUBBER STOPPER

STERILE STEEL WIRE USED TO SUSPEND STUD

GLASS BOTTLE, 125cm3

CAPACITY

TEST BOTTLE SOLUTION:ANAROBIC, BOTTLE FILLEDTO NECK

RUBBER ‘O’ RING

MILD STEEL STUD FROM BIOFILM GENERATOR

PLASTIC COATED STIRRINGBAR

MAGNETIC STIRRER

bacteria and biocide

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