EI Research Report Investigation of microbiological susceptibility of biodiesel and biodiesel blends
EI RESEARCH REPORT INVESTIGATION OF MICROBIOLOGICAL SUSCEPTIBILITY OF BIODIESEL AND BIODIESEL BLENDS March 2014 Published by ENERGY INSTITUTE, LONDON The Energy Institute is a professional membership body incorporated by Royal Charter 2003 Registered charity number 1097899
The Energy Institute (EI) is the chartered professional membership body for the energy industry, supporting over 19 000 individuals working in or studying energy and 200 energy companies worldwide. The EI provides learning and networking opportunities to support professional development, as well as professional recognition and technical and scientific knowledge resources on energy in all its forms and applications. The EI s purpose is to develop and disseminate knowledge, skills and good practice towards a safe, secure and sustainable energy system. In fulfilling this mission, the EI addresses the depth and breadth of the energy sector, from fuels and fuels distribution to health and safety, sustainability and the environment. It also informs policy by providing a platform for debate and scientifically-sound information on energy issues. The EI is licensed by: the Engineering Council to award Chartered, Incorporated and Engineering Technician status; the Science Council to award Chartered Scientist status, and the Society for the Environment to award Chartered Environmentalist status. It also offers its own Chartered Energy Engineer, Chartered Petroleum Engineer and Chartered Energy Manager titles. A registered charity, the EI serves society with independence, professionalism and a wealth of expertise in all energy matters. This publication has been produced as a result of work carried out within the Technical Team of the EI, funded by the EI s Technical Partners. The EI s Technical Work Programme provides industry with cost-effective, value-adding knowledge on key current and future issues affecting those operating in the energy sector, both in the UK and internationally. For further information, please visit http://www.energyinst.org The EI gratefully acknowledges the financial contributions towards the scientific and technical programme from the following companies BG Group Premier Oil BP Exploration Operating Co Ltd RWE npower BP Oil UK Ltd Saudi Aramco Centrica Scottish Power Chevron SGS ConocoPhillips Ltd Shell UK Oil Products Limited Dana Petroleum Shell U.K. Exploration and Production Ltd DONG Energy SSE EDF Energy Statkraft ENI Statoil E. ON UK Talisman Energy (UK) Ltd ExxonMobil International Ltd Total E&P UK Limited International Power Total UK Limited Kuwait Petroleum International Ltd Tullow Maersk Oil North Sea UK Limited Valero Murco Petroleum Ltd Vattenfall Nexen Vitol Phillips 66 World Fuel Services However, it should be noted that the above organisations have not all been directly involved in the development of this publication, nor do they necessarily endorse its content. Copyright 2014 by the Energy Institute, London. The Energy Institute is a professional membership body incorporated by Royal Charter 2003. Registered charity number 1097899, England All rights reserved No part of this book may be reproduced by any means, or transmitted or translated into a machine language without the written permission of the publisher. ISBN 978 0 85293 658 0 Published by the Energy Institute The information contained in this publication is provided for general information purposes only. Whilst the Energy Institute and the contributors have applied reasonable care in developing this publication, no representations or warranties, express or implied, are made by the Energy Institute or any of the contributors concerning the applicability, suitability, accuracy or completeness of the information contained herein and the Energy Institute and the contributors accept no responsibility whatsoever for the use of this information. Neither the Energy Institute nor any of the contributors shall be liable in any way for any liability, loss, cost or damage incurred as a result of the receipt or use of the information contained herein. Further copies can be obtained from: Portland Customer Services, Commerce Way, Whitehall Industrial Estate, Colchester CO2 8HP, UK. t: +44 (0)1206 796 351 e: sales@portland-services.com Electronic access to EI and IP publications is available via our website, www.energypublishing.org. Documents can be purchased online as downloadable pdfs or on an annual subscription for single users and companies. For more information, contact the EI Publications Team. e: pubs@energyinst.org
CONTENTS Page Foreword...3 Acknowledgements...4 1 Introduction...5 1.1 Technical background...5 1.1.1 FAME in automotive diesel...5 1.1.2 FAME in aviation kerosene...6 1.2 Project scope...6 2 Test methods....7 2.1 Set-up of microcosms...7 2.1.1 FAME...7 2.1.2 Diesel base fuel...7 2.1.3 Aviation fuel....7 2.1.4 Screening blend components for biocidal activity and microbial contamination....7 2.1.5 Preparation of fuel blends...8 2.1.6 Test micro-organisms...8 2.1.7 Test microcosms...10 2.2 Assessment of test microcosms...10 2.2.1 Routine assessment...10 2.2.2 Procedure for sampling test microcosms....12 2.2.3 Additional assessments at the end of the trial....12 3 Results...14 3.1 Diesel fuel study...14 3.1.1 Visual appearance of diesel microcosms...14 3.1.2 ph of aqueous phase of diesel microcosms....14 3.1.3 Total viable counts of aqueous phase of diesel microcosms....15 3.1.4 MicrobMonitor2 tests of aqueous and fuel phase of diesel microcosms..18 3.1.5 ATP measurements of aqueous and fuel phase of diesel microcosms (ASTM D7463)...21 3.1.6 Total viable counts of fuel phase of diesel microcosms (IP 385)... 23 3.1.7 Gravimetric and particulates in fuel and interface phase of diesel microcosms (modified IP 415)... 23 3.1.8 Water content of fuel phase of diesel microcosms (IP 438)............ 24 3.1.9 Identification of microbial isolates in diesel microcosms...25 3.2 Aviation kerosene study....28 3.2.1 Visual appearance of aviation kerosene microcosms...28 3.2.2 ph of aqueous phase of aviation kerosene microcosms...28 3.2.3 Total viable counts of aqueous phase of aviation kerosene microcosms...29 3.2.4 MicrobMonitor2 tests of aqueous and fuel phase of aviation kerosene microcosms...30 3.2.5 ATP measurements of aqueous and fuel phase of aviation kerosene microcosms (ASTM D7463)... 32 3.2.6 Total viable counts of fuel phase of aviation kerosene microcosms (IP 385) 34 3.2.7 Gravimetric and particulates in fuel and interface phase of aviation kerosene microcosms (modified IP 423)... 34 1
3.2.8 Water content of fuel phase of aviation kerosene microcosms (IP 438)... 35 3.2.9 Identification of microbial isolates in aviation kerosene microcosms...36 4 Discussion and conclusions...38 4.1 Diesel fuel study...38 4.2 Aviation fuel study....39 Annexes Annex A Test micro-organisms....40 Annex B Photographs of test microcosms...44 Annex C Results of microbiological assays...58 Annex D References....65 2
FOREWORD The petroleum industry has raised concerns that the increasingly widespread use of Fatty Acid Methyl Esters (FAME) in road transport and other diesel fuels will make fuels more susceptible to microbial spoilage and associated operational problems during their distribution and use. FAME is more readily biodegraded than mineral fuels, and anecdotal evidence from the field suggests a significant increase in operational problems as a result of microbial growth and contamination of diesel fuel systems. These problems include increased plugging of fuel filters at diesel retail sites and in large diesel vehicles, and also microbially-influenced corrosion of fuel tanks. There has been consequent speculation that if FAME inadvertently contaminates aviation fuels during transport, for example, in non-dedicated pipelines or ship tankers, there may also be increased microbial growth in aviation fuel tanks and systems, with potentially dramatic impact on aircraft operational safety. The Energy Institute (EI) commissioned a review paper, Implications of biofuels on microbial spoilage and corrosion within the fuel distribution chain and end use, published in May 2008, which identified that very little research had been conducted to establish the extent to which FAME content influenced the susceptibility of fuels to microbial spoilage. At the request of the EI s Microbiology Committee, a laboratory study was undertaken to investigate the influence of FAME concentration on the susceptibility of fuels to microbiological spoilage. The study consisted of two parts: one investigated the influence of various FAME blend concentrations (2 % (B2) to 100 % (B100)) in diesel fuel; and the second investigated the influence of trace concentrations (100 ppm and 400 ppm) of FAME in aviation fuel. Microcosms of zero-sulphur diesel (ZSD) and high sulphur diesel (HSD) fuel blended with FAME were inoculated with a mixture of fuel spoilage micro-organisms in a small amount of aqueous phase and were assessed for the extent of subsequent microbial growth by a number of techniques. A similar study was conducted for both mercaptan oxidation (MEROX)-treated and hydro-treated aviation kerosene without FAME and with FAME at 100 ppm and 400 ppm. In both studies the rate and extent of microbial growth was compared to the base fuels without FAME. The opportunity was also used to compare a number of different methods for assessing microbial contamination in fuels, including commercially available test kits which have been adopted, or are under consideration, as industry standard methods. This research report presents the results of the laboratory study. It concludes that FAME does increase the susceptibility of fuels to microbial growth, most notably fungal growth. Under the test conditions, the increase in susceptibility of the fuels to microbial growth increased with increasing FAME concentration. For diesel fuels it is concluded that when FAME is blended above 2 %, the influence on the rate and extent of microbial growth warrants additional consideration in the maintenance and monitoring of fuel handling facilities. For aviation fuels it is concluded that, although FAME did marginally increase susceptibility to microbial growth, the influence was modest compared to the influence of other factors associated with MEROX treatment as opposed to hydro-treatment. At the concentrations proposed as de minimus limit values in aviation kerosene (provisionally 100 ppm), any increased susceptibility to microbial growth due to the presence of FAME is unlikely to have significant operational impact. 3
ACKNOWLEDGEMENTS This project was commissioned by the Energy Institute s Microbiology Committee. The work was carried out by Graham Hill, ECHA Microbiology Limited, United Kingdom and steered by members of the Microbiology Committee, who during the project included: Simon Ashton Simon Christopher Brian Crook Carol Devine Bob Eden Beate Hildenbrand Graham Hill Joan Kelley Jan Kuever Jan Larsen Bart Lomans Torben Lund Skovhus Elaine McFarlane Andrew Price Tony Rizk Kerry Sinclair Jim Stott Ian Vance Neil Whitehead ExxonMobil BP HSL North East Corrosion Engineers (NECE) RawWater Engineering Company Energy Institute ECHA Microbiology CABI UK Bremen Institute for Materials Testing Maersk Oil & Gas Shell Det Norske Veritas (DNV) Shell Global Solutions Oil Plus Saudi Aramco Energy Institute Intertek - Capcis Centromere Minton Treharne & Davies Ltd The EI wishes to record its appreciation of the work carried out by the author and also its gratitude for the valuable contributions made by the Microbiology Committee during the course of the project. 4
1 INTRODUCTION 1.1 TECHNICAL BACKGROUND 1.1.1 FAME in automotive diesel The European Union Directive 2003/30/EC on the promotion of the use of biofuels or other renewable fuels for transport, mandates that all fuel sold for use in road vehicles should contain a minimum 5,75 % (calculated on the basis of energy content) of components derived from renewable sources. For automotive diesels the requirement is implemented in practice by blending conventional fossil diesel with Fatty Acid Methyl Esters (FAME) derived from a variety of plant sources; including rape seed oil, soy bean oil, palm oil and also from animal fats (tallow). Member states have shown varying degrees of progress towards meeting the Directives requirements, but in much of Europe automotive diesel has for the past few years contained in the order of 2-5 % FAME. These concentrations are likely to increase in the foreseeable future and in some countries diesel containing 7 or 10 % FAME is already widely used. Automotive diesel specification EN590 currently allows up to 7 % FAME. Industry experience suggests that diesel fuels containing FAME can have an increased susceptibility to microbial growth (Hill and Hill, 2009). The Energy Institute (EI) published a review paper (Price, 2008) which highlighted the need for further research. The EI has also issued a Technical Bulletin which discusses the implications of FAME on microbial growth and provides provisional recommendations for the maintenance of fuel handling facilities. The Technical Bulletin expands on previously issued technical guidance already published by the EI for all fuel types (Hill, 2008). Microbial growth by bacteria and fungi in diesel storage tanks, distribution facilities and end-user tanks can lead to contamination of the diesel with microbial particulates (biomass) which can cause severe filter blocking problems and blocking of fuel lines. Significant operational issues have already been experienced by some fuel retailers, where rapid onset of pump filter blocking has been encountered. Fuel filter blocking and fuel starvation problems have also been experienced by some truck and bus fleets; although, to date, operational problems attributed to microbial contamination of diesel in private cars have been extremely rare. Field evidence suggests that the most extensive problems associated with microbial growth occur in diesel where FAME is blended at less than 20 % but greater than 2 %. There is some evidence that when FAME blend concentration exceeds approx. 20 % in diesel, providing excessive amounts of visible water, then microbial growth may actually be reduced or inhibited. This is because FAME has a water scavenging effect. Microbes can only grow when free water is present; when high concentrations of FAME are present, water is dissolved in the fuel and is no longer available for microbial growth. This document reports research on the influence of FAME concentration on susceptibility of diesel to microbial growth. On-going research will address in more detail the influence of water content. 5