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Aircraft manufacturers are even more cautious due to the inherent risks of air travel, but a test flight has been performed by a Czech Aircraft (completely powered on biofuel); testing has been announced by Rolls Royce plc, Air New Zealand and Boeing (one engine out of four on a Boeing 747); and commercial passenger jet testing has also been announced by Virgin Atlantic's Richard Branson.

The world's first biofuel-powered commercial aircraft took off from London's Heathrow Airport on February 24, 2008 and touched down in Amsterdam on a demonstration flight hailed as a first step towards "cleaner" flying. The "BioJet" fuel for this flight was produced by Seattle based Imperium Renewables, Inc.

The world's first commercial aviation test flight powered by the sustainable second-generation biofuel jatropha has been successfully completed in Auckland. More than a dozen key performance tests were undertaken in the two hour test flight which took-off at 11:30am (NZ time), 30th December 2008 from Auckland International Airport. A biofuel blend of 50:50 jatropha and Jet A1 fuel was used to power one of the Air New Zealand Boeing 747-400's Rolls-Royce RB211 engines.

The test flight is a joint initiative between Air New Zealand, Boeing, Rolls-Royce and Honeywell's UOP, with support from Terasol Energy. The jatropha oil Air New Zealand sourced and refined for its test flight came from South Eastern Africa (Malawi, Mozambique and Tanzania) and India. It was sourced from seeds grown on environmentally sustainable farms. Once received from Terasol Energy, the jatropha oil was refined through a collaborative effort between Air New Zealand, Boeing and leading refining technology developer UOP, utilising UOP technology to produce jet fuel from renewable sources that can serve as a direct replacement to traditional petroleum-based fuel.

Biodiesel can also be used as a heating fuel in domestic and commercial boilers, sometimes known as bioheat. Older furnaces may contain rubber parts that would be affected by biodiesel's solvent properties, but can otherwise burn biodiesel without any conversion required. Care must be taken at first, however, given that varnishes left behind by petro diesel will be released and can clog pipes- fuel filtering and prompt filter replacement is required. Another approach is to start using biodiesel as blend, and decreasing the petroleum proportion over time can allow the varnishes to come off more gradually and be less likely to clog.

Thanks to its strong solvent properties, however, the furnace is cleaned out and generally becomes more efficient. A technical research paper describes laboratory research and field trials project using pure biodiesel and biodiesel blends as a heating fuel in oil fired boilers. During the Biodiesel Expo 2006 in the UK, Andrew J. Robertson presented his biodiesel heating oil research from his technical paper and suggested that B20 biodiesel could reduce UK household CO2 emissions by 1.5 million tons per year.

The British businessman Richard Branson's Virgin Voyager train, number 220007 Thames Voyager, billed as the world's first "biodiesel train" was converted to run on 80% petro diesel and only 20% biodiesel, and it is claimed it will save 14% on direct emissions.

Similarly, a train in eastern Washington will be running on a 25% biodiesel 75% petro diesel blend during summer, purchasing fuel from a biodiesel producer seated along the railroad tracks. The train will be powered by biodiesel made in part from Washington-grown canola.