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MARINE BIOREMEDIATION TECHNOLOGIES SCREENING
MATRIX AND REFERENCE GUIDE
Cádiz, July 2002
Edited by
Prof. Dr. Jose A. Perales Vargas-Machuca
Prof. Dr. Manuel A. Manzano Quiñones
Environmental Technology Research Group
University of Cádiz
Spain
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
NATURAL ATTENUATION AND MONITORING
OIL ON SHORE
IS THERE A SAFEACCESS?
PROTECT THREATENED
AREAS.MECHANICAL
RECOVERY FROM THE SEA WHEN THE
OIL MOVES FROM THE COAST
NON BIOLOGICAL
IN SITU TECHNIQUES
IS AN EX SITU TECHNIQUE
FEASIBLE AND EFFECTIVE?
IS THERE A THREATENED
SITE?
IS FREE OIL PRESENT?
IS A NON BIOLOGICAL IN
SITU TECHNIQUE FEASIBLE AND EFFECTIVE ?
IS IT A HIGH ENERGY COAST?
COMPLETED OBJECTIVES?
EROSION RISK?
EX SITU BIOLOGICAL TECHNIQUES
ON-SITE CHEMICAL -PHYSICAL
TREATMENT
IS THE OILBIODEGRADABLE?
IS IT A SITE OF SPECIAL CONCERN?
(AMENITY BEACH OR ECOLOGICAL IMPLICATIONS)
IS THE OILBIODEGRADABLE?
IS OXYGEN CONCENTRATION
ADEQUATE?
MONITORING
AERATE
IS AERATION FEASIBLE?
CAN WE WAIT UNTIL WARMER SEASONS?
IS THE TEMPERATURE
ADEQUATE?
ADD NUTRIENTSIS NUTRIENT CONTENT
SUFFICIENT?
OK
YESNO
YES
YES
YES
YES
NO
NO
NO
NO
NO
NO
YES
NO
NO
NO
NO
NO
YES
NO
YES
YES
YES
YES
YES
YES
YES
NO
YES
YES
DETERMINING CRITERIA FOR THE OBJECTIVES
RECONSIDERER OTHER OBJECTIVES?
IS THERE OTHER LIMITING FACTOR?NO
YES
OBJECTIVES COMPLETED?
OK
YES
NO
NATURAL ATTENUATION AND MONITORING
OIL ON SHORE
IS THERE A SAFEACCESS?
PROTECT THREATENED
AREAS.MECHANICAL
RECOVERY FROM THE SEA WHEN THE
OIL MOVES FROM THE COAST
NON BIOLOGICAL
IN SITU TECHNIQUES
IS AN EX SITU TECHNIQUE
FEASIBLE AND EFFECTIVE?
IS THERE A THREATENED
SITE?
IS FREE OIL PRESENT?
IS A NON BIOLOGICAL IN
SITU TECHNIQUE FEASIBLE AND EFFECTIVE ?
IS IT A HIGH ENERGY COAST?
COMPLETED OBJECTIVES?
EROSION RISK?
EX SITU BIOLOGICAL TECHNIQUES
ON-SITE CHEMICAL -PHYSICAL
TREATMENT
IS THE OILBIODEGRADABLE?
IS IT A SITE OF SPECIAL CONCERN?
(AMENITY BEACH OR ECOLOGICAL IMPLICATIONS)
IS THE OILBIODEGRADABLE?
IS OXYGEN CONCENTRATION
ADEQUATE?
MONITORING
IN-SITU BIOSTIMULATION BY ADDITION OF OXYGEN
IS AERATION FEASIBLE?
CAN WE WAIT UNTIL WARMER SEASONS?
IS THE TEMPERATURE
ADEQUATE?
IN-SITU BIOSTIMULATION WITH NUTRIENTS
IS NUTRIENT CONTENT SUFFICIENT?
OK
YESNO
YES
YES
YES
YES
NO
NO
NO
NO
NO
NO
YES
NO
NO
NO
NO
NO
YES
NO
YES
YES
YES
YES
YES
YES
YES
NO
NO
YES
YES
DETERMINING CRITERIA FOR THE OBJECTIVES
RECONSIDERER OTHER OBJECTIVES?
IS THERE OTHER LIMITING FACTOR?NO
YES
OBJECTIVES COMPLETED?
OK
YES
NO
Return
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
OIL ON SHORE
Return to Flowchart Back to the previous slide Explanations
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
DETERMINING CRITERIA FOR THE OBJECTIVES
Return to Flowchart Back to the previous slide Explanations
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS THERE A SAFE ACCESS?
YES
Return to Flowchart Back to the previous slide Explanations
NO
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS FREE OIL PRESENT?
Return to Flowchart Back to the previous slide Explanations
YES NO
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
RECONSIDER OTHER OBJECTIVES
Return to Flowchart Back to the previous slide
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS THERE A THREATENED SITE?
Return to Flowchart Back to the previous slide Explanations
YES NO
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
PROTECT THREATENED AREAS. MECHANICAL RECOVERY
FROM THE SEA WHEN THE OIL MOVES
FROM THE COAST
Return to Flowchart Back to the previous slide
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS IT A HIGH ENERGY COAST?
Return to Flowchart Back to the previous slide Explanations
YES NO
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS IT A SITE OF SPECIAL CONCERN ?
Return to Flowchart Back to the previous slide Explanations
YES NO
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS THE OIL BIODEGRADABLE ?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS A NON BIOLOGICAL IN-SITU TECHNIQUE
FEASIBLE AND EFFECTIVE?
YES NO
Return to Flowchart Back to the previous slide
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
NON BIOLOGICAL IN-SITU TECHNIQUES
Return to Flowchart Back to the previous slide
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
COMPLETED OBJECTIVES?
YES NO
Return to Flowchart Back to the previous slide
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
TREATMENT FINISHED
Home Back to the previous slide
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS THE TEMPERATUREADEQUATE?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
CAN WE WAIT UNTIL WARMER SEASONS?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS OXYGEN CONCENTRATION ADEQUATE?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS AERATION FEASIBLE?
YES NO
Return to Flowchart Back to the previous slide
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IN-SITU BIOREMEDIATION BY OXYGEN ADDITION TECHNIQUES
Return to Flowchart Back to the previous slide
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS AN EX-SITU TECHNIQUE FEASIBLE AND EFFECTIVE?
YES NO
Return to Flowchart Back to the previous slide
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
EROSION RISK?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS THE OIL BIODEGRADABLE ?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
ON SITE CHEMICAL-PHYSICALTREATMENT
Return to Flowchart Back to the previous slide
Next
Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
EX-SITU BIOLOGICAL TECHNIQUES
Return to Flowchart Back to the previous slide
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS NUTRIENT CONTENT SUFFICIENT ?
YES NO
Return to Flowchart Back to the previous slide Explanations
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IN-SITU BIOREMEDIATION BY NUTRIENT ADDITION TECHNIQUES
Return to Flowchart Back to the previous slide
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
MONITORING
Return to Flowchart Back to the previous slide Explanations
Next
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
COMPLETED OBJECTIVES?
YES NO
Return to Flowchart Back to the previous slide
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
IS THERE OTHER LIMITING FACTOR ?
YES NO
Return to Flowchart Back to the previous slide Explanations
SEE NOTE
MARINE BIOREMEDIATION TECHNOLOGIES SCREENING MATRIX
AND REFERENCE GUIDE
NATURAL ATTENUATION AND MONITORING
Return to Flowchart Back to the previous slide
Next
Back to the previous slide
Oil on Shore
This is a bioremediation guide only for oil and marineenvironments. On the other hand, bioremediation has beenconsidered as an effective technology only in the sediment.Therefore, the starting point of the flowchart will be the presenceof crude oil on the shore.
Back to the previous slide
Determining Criteria for Objectives
The first consideration has to be the establishment the reason forbioremediation, i.e., ecological and economic implicationsbecause they will determine not only the technology to be usedbut also the required final oil concentration
Back to the previous slide
Is there safe access?
In occasions and due to the particular morphology of thecoast, for example wave-cut rock platforms it becomes verydifficult or practically impossible to carry out bioremediationtechnologies. In this situations the only solution is naturalattenuation and monitoring.
Back to the previous slide
Is free oil present?
Bioremediation is considered the second phase of the plan,first being the removal of free oil. Therefore, if the shoreline issaturated with oil it is first necessary and very important to removeit. The presence of free oil on shoreline can be easily noticed bysimple visual observation.
Back to the previous slide
Is there a threatened site?
Before consider which clean-up technologies should beused, it is necessary to know if the oil spill is still spreading toothers non contaminated areas. In this case, the first actionshould be to confine the oil.
Back to the previous slide
Is it a high energy coast?
It is very important to differentiate between high and low energycoasts in order to determine what type of bioremediationtechnology is the most appropriate for each case.
Generally in high energy coasts in-situ bioremediationtechnologies are not feasible because the sediments are likely tobe removed by tidal action.
In order to have information about the classification of the offshorerelated with its energy, geomorphologic cartography is a veryuseful tool.
Back to the previous slide
Is it a site of special concern?(Amenity beach or ecological implications)
One important question is how long will clean-up take. Related tothis aspect, in-situ bioremediation technologies are long-termclean-up technologies, so they can not be selected when in ahurry are which is the case with economic (amenity beach) orecological reasons (high ecological value zone). Nevertheless, itis necessary to keep in mind that as fast is a clean-up technology,higher is the environmental impact that its application couldprovoke on the contaminated compartment, so a balance betweenboth possibilities, impact from the oil spill and impact from carryout the cleaning process, should be considered.
Back to the previous slide
Is oil biodegradable?
Bioremediation is only applicable in significantly biodegradable oils. Farfrom being a homogeneous substance, crude oil is a complex mixture ofthousands of different chemical compounds. Since components ofpetroleum degrade at different rates, it is difficult and misleading tospeak in terms of an overall biodegradation rate.Biodegradation rates are typically highest for the saturates, followed bythe light aromatics, with high-molecular-weight aromatics, asphaltenes,and resins exhibiting extremely low rates of degradation. As a spillweathers, its composition changes: the light aromatics and light alkanesdissolve or evaporate rapidly and are metabolised by microorganisms.The heavier components that are harder to degrade remain. Thereforeno crude oil is subject to complete biodegradation, and claims that all ofa light oil or more than 50 percent of a heavy oil can be biodegraded indays or weeks are highly suspect.
Back to the previous slide
Is the temperature adequate?
At low temperature, the rate of hydrocarbon metabolism bymicroorganisms decreases. Also, lighter fractions of petroleumbecome less volatile, thereby leaving the petroleum constituentsthat are toxic to microbes in the water for a longer time anddepressing microbial activity. Petroleum also becomes moreviscous at low temperature. Hence, less spreading occurs andless surface area is available for colonisation by microorganisms.Therefore, a minimum temperature is required in order to promotesuch bio-reactions. This minimum temperature is between 5 and10ºC, depending on the latitude.
Back to the previous slide
Can we wait until warmer seasons?
In temperate regions, seasonal changes in water temperatureaffect the rate of biodegradation, but the process continues year-round.
Back to the previous slide
Is oxygen concentration adequate?
Oxygen is one of the most important requirements for microbial degradation ofhydrocarbons. When oxygen is not available, the rates of biodegradation decrease.Thus, oil that has sunk to the sea floor and been covered by sediment takes muchlonger to degrade. Oxygen availability there is determined by depth in the sediment,height of the water column, and turbulence. Low-energy beaches and fine-grainedsediments may also be depleted in oxygen; thus, the rate of biodegradation may belimited in these areas.
There are two organoleptic indicators that can be easily used to establish if a sedimentis under anaerobic or aerobic conditions. One is the colour of the sediment, which isblack under anaerobic conditions due to formation of metallic sulfides. Nevertheless, thisindicator could not be very useful because the colour of the oil. The second indicator isthe odour of the anaerobic sediment, which is nauseating due to emissions of hydrogensulfide acids, ammonia and intermediates from degradation of organic compounds withnitrogen and sulfur.
Back to the previous slide
Is the nutrient content sufficient?
Nutrients such as nitrogen and phosphorus play a critical role in limitingthe rate of biodegradation in marine waters. Several studies haveshown that an inadequate supply of these nutrients may result in a slowrate of biodegradation. Although petroleum is rich in the carbon requiredby microorganisms, it is deficient in the mineral nutrients necessary tosupport microbial growth. Marine and other ecosystems are oftendeficient in these substances because non-oil degradingmicroorganisms (including phytoplankton) consume them in competitionwith the oil degrading species. Also, phosphorus precipitates as calciumphosphate at the pH of seawater. The nutritional requirements of carbonto nitrogen is 10:1 and carbon to phosphorus 30:1.
Back to the previous slide
Monitoring
For proper application of the technology, there is a need for monitoringprogrammes to quantify intrinsic rates of oil loss and degradation, demostratetreatment efficacy and identify operational endpoint.
A major obstacle is heterogeneity within the natural environment. Absolutelevels of contamination can vary widely over a site and simple estimates ofbiodegradation based on sequential samples can be confounded by thisheterogeneity, unless large number of samples are taken. This problem can beresolved by the normalisation of data to conserved markers such us hopanesand chrysenes found within the oil. Nevertheless this solution has high costsand are time consuming. Other possibilities include reliable microbial responseand ecotoxicologycal monitoring protocols to verify efficacy for toxicity reductionover that of no treatment.
Back to the previous slide
Are there other limiting factors?
In addition to the nutrients just cited, some organisms mayalso need organic nutrients. Required organic nutrients,know as growth factors, are compounds needed byorganisms as precursors or constituents of organic cellmaterial that cannot be synthesised from other carbonsources. Although growth factors requirements differ from oneorganism to another, the major growth factors fall into thefollow three main classes: amino acids, purines andpyrimidines and vitamins.
Back to the previous slide
Are there other limiting factors?
Note: If at the previous question about if the oxygenconcentration is adequate the answer was not, then it has tobe considered that the oxygen is a limiting factor.
Back to the previous slide
On-site Chemical-Physical Treatment
The on-site treatments consists on those ex-situ technologieswhere the removed sediment is located nearby the contaminatedarea but in a zone not influenced by the tide.
Back to the previous slide
Erosion Risk
If an ex-situ bioremediation technology is applied, the removal ofthe original sediments can affect the littoral drift, because itcauses changes in the granuloumetry of the sediment
