ASTM D7978 - 14

    Standard Test Method for Determination of the Viable Aerobic Microbial Content of Fuels and Associated Water—Thixotropic Gel Culture Method

    Active Standard ASTM D7978 | Developed by Subcommittee: D02.14

    Book of Standards Volume: 05.05


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    Significance and Use

    5.1 This test method is intended to provide a tool for assessing whether fuel storage and distribution facilities or end user fuel tanks are subject to microbial growth and alert fuel suppliers or users to the potential for fuel quality or operational problems and/or the requirement for preventative or remedial measures.

    5.2 This test method detects numbers of microbial colony forming units (CFU), the same detection parameter used in the laboratory standard procedures Practice D6974 and IP 385. However, whereas Practice D6974 and IP 385 provide separate assessment of numbers of viable aerobic bacteria CFU and numbers of viable fungal CFU, this test method provides a combined total count of viable aerobic bacteria and fungal CFU.

    5.3 This test method is designed to detect a recognized group of microorganisms of significance in relation to contamination of distillate fuels, but it is recognized that microbiological culture techniques do not detect all microorganisms that can be present in a sample. Culturability is affected primarily by the ability of captured microbes to proliferate on the growth medium provided, under specific growth conditions. Consequently, a proportion of the active or inactive microbial population present in a sample can be viable but not detected by any one culture test.7 In this respect, the test is indicative of the extent of microbial contamination in a sample ,and it is assumed that when a fuel sample is significantly contaminated, some of the dominant microbial species present will be quantifiably detected, even if not all species present are culturable.

    5.4 Many samples from fuel systems can be expected to contain a low level of “background” microbial contamination, which is not necessarily of operational significance. The minimum detection level of this test method is determined by the volume of specimen tested and is set such that microbial contamination will generally only be detected when it is at levels indicative of active proliferation.

    5.5 The test will detect culturable bacteria and fungi that are metabolically active and dormant fungal spores. Presence of fungal spores in a fuel sample can be indicative of active microbial proliferation within a fuel tank or system, but at a point distant from the location sampled. Active microbial growth only occurs in free water, and this can be present only as isolated pockets at tank or system low points. Because fungal spores are more hydrophobic than active cells and fungal material (mycelium), they disperse more readily in fuel phase and are thus more readily detected when low points cannot be directly sampled and only fuel phase is present in samples.

    5.6 This test method can determine whether microbial contamination in samples drawn from fuel tanks and systems is absent or present at light, moderate, and heavy levels.

    5.7 The categorization of light, moderate, and heavy levels of contamination will depend on the fuel type, the sampling location, the facility sampled, and its specific operating circumstances.

    5.8 Further guidance or interpretation of test results can be found in Guide D6469, in the Energy Institute Guidelines for the investigation of the microbial content of petroleum fuels, and for the implementation of avoidance and remedial strategies and in the IATA Guidance Material on Microbiological Contamination in Aircraft Fuel Tanks.

    5.8.1 Further guidance on sampling can be found in Practice D7464.

    5.9 Testing can be conducted on a routine basis or to investigate incidents.

    5.10 Microbiological tests are not intended to be used to determine compliance with absolute fuel specifications or limits. The implementation of specification limits for microbiological contamination in fuels is generally not appropriate, and microbial contamination levels cannot be used alone or directly to make inferences about fuel quality or fitness for use.

    5.11 When interpreting results, it must be appreciated that the test result applies only to the specific sample and specimen tested and not necessarily to the bulk fuel. Microbiological contamination usually shows a highly heterogeneous distribution in fuel systems, and therefore, analysis of a single sample will rarely provide a complete assessment of the overall levels of contamination present.

    5.12 Water phase will usually contain substantially higher numbers of microbial CFU than fuel phase and, consequently, a different interpretation of results is required.

    1. Scope

    1.1 This test method describes a procedure that can be used in the field or in a laboratory to quantify culturable, viable aerobic microorganisms present as contaminants in liquid fuels, including those blended with synthesized hydrocarbons or biofuels, with kinematic viscosities (at 40 °C) of 24 mm2 s-1 and heavy and residual fuels with kinematic viscosities (at 40 °C) of 700 mm2 s-1 and in fuel-associated water.

    1.1.1 This test method has been validated by an ILS for a range of middle distillate fuels meeting Specifications D975, D1655, ISO 8217 DMA, and NATO F-76.2

    1.2 This test method quantitatively assesses culturable, viable aerobic microbial content present in the form of bacteria, fungi, and fungal spores. Results are expressed as the total number of microbial colony forming units (CFU)/L of fuel or total number of CFU/mL of associated water. The number of CFU should not be interpreted as absolute values but should be used as part of a diagnostic or condition monitoring effort; for example, these values can be used to assess contamination as absent, light, moderate, or heavy.

    Note 1: This test method is technically equivalent to IP 613, although the two methods are not currently jointed.

    1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.

    1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.


    2. Referenced Documents (purchase separately) The documents listed below are referenced within the subject standard but are not provided as part of the standard.

    ASTM Standards

    D975 Specification for Diesel Fuel Oils

    D1129 Terminology Relating to Water

    D1655 Specification for Aviation Turbine Fuels

    D4175 Terminology Relating to Petroleum, Petroleum Products, and Lubricants

    D4176 Test Method for Free Water and Particulate Contamination in Distillate Fuels (Visual Inspection Procedures)

    D6469 Guide for Microbial Contamination in Fuels and Fuel Systems

    D6974 Practice for Enumeration of Viable Bacteria and Fungi in Liquid Fuels--Filtration and Culture Procedures

    D7464 Practice for Manual Sampling of Liquid Fuels, Associated Materials and Fuel System Components for Microbiological Testing

    D7847 Guide for Interlaboratory Studies for Microbiological Test Methods

    E2756 Terminology Relating to Antimicrobial and Antiviral Agents

    Energy Institute Standards

    Guidelines for the Investigation of the Microbial Content of Petroleum Fuels and for the Implementation of Avoidance and Remedial Strategies, 2nd Edition, 2008, Energy Institute, London, ISBN 978 0 85293 524 8

    IP 385 Determination of the Viable Aerobic Microbial Content of Fuels and Fuel Components Boiling Below 390 C--Filtration and Culture Method

    IP 613 Determination of the Viable Aerobic Microbial Content of Fuels and Associated Water--Thixotropic Gel Culture Method

    Other Standards

    IATA Guidance Material on Microbiological Contamination in Aircraft Fuel Tanks, 4th Edition, December 2011 Available from International Air Transport Assocation (IATA), 800 Place Victoria, PO Box 113, Montreal H4Z 1M1, Quebec, Canada or 33, Route de lAeroport, PO Box 416, 1215 Geneva 15 Airport, Switzerland, http://www.iata.org.

    ISO 8217 DMA Petroleum Products, Fuels (Class F), Specifications of Marine Fuels, 4th Edition 2010, ISO Switzerland


    ICS Code

    ICS Number Code 75.160.20 (Liquid fuels)

    UNSPSC Code

    UNSPSC Code 77121700(Water pollution)


    Referencing This Standard
    Link Here
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    DOI: 10.1520/D7978-14

    Citation Format

    ASTM D7978-14, Standard Test Method for Determination of the Viable Aerobic Microbial Content of Fuels and Associated Water—Thixotropic Gel Culture Method, ASTM International, West Conshohocken, PA, 2014, www.astm.org

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