Microbial diversity in trace water of jet fuel in Vietnam

Transcription

1 Title Authr(s) Citatin Micrbial diversity in trace water f jet fuel in Vietnam Lai, Thuy Hien; Pham, Thi Hang; Vung, Thi Nga; Hai, Hang; Yamaguchi, Nbuyasu; Tani, Katsuji Annual Reprt f FY 2006, The Cre University Prgram between Japan Sciety fr the Prmtin f Science (JSPS) and Vietnamese Academy f Science and Technlgy (VAST). P.413-P.420 Issue Date 2007 Text Versin publisher URL DOI rights

2 Micrbial diversity in trace water f jet fuel in Vietnam Lai Thuy Hien l,*, Pham Thi Hangl, Vung Thi Nga l, Hang Hae, Nbuyasu Yamaguchi 3, Katsuji Tani 3 1 Department 0/ Petrleum Micrbilgy, Institute /Bitechnlgy, VAST, Hani, Vietnam 2Divisin /Sustainable Energy and Envirnmental Engineering, Graduate Schl 0/ Engineering, Osaka University, 2-1 Yamadaka, Suita, Osaka , Japan 3Graduate Schl /Pharmaceutical SCiences, Osaka University, 1-6 Yamadaka, Suita, Osaka , Japan Keywrds: Aircraft fuel, micrbial diversity, jet AI-hydrcarbn using bacteria Intrductin Micrrganisms in petrleum prduct, including jet fuel, have been fund by sme English and American authrs since the 20's f the last century. In 1958, after Bakanaukas islated many micrrganisms in jet fuel JP4, this tpic was especially cncentrated. Mst f micrrganisms living in jet fuel are able t degrade hydrcarbn f the fuel t btain energy and carbn surce fr their grwth. The presence f micrrganisms in jet fuel nt nly brings abut the lst f fuel in stre prcess, but als briefly changes the cmpnent and structure f jet fuel. Mre ver, the prducts f their metablism, such as rganic acids, alchls, H2S... are crrsive. Hence, they cause many prblems fr fuel reservirs and tanks, leak plugging pipelines. Besides, filter system and plugging pipeline can becme bstructed because f micrrganisms' bimass. This influences the plane safety. Therefre, many cuntries in the wrld cnsider micrrganism criteria as an imprtant standard t estimate the jet fuel quality. Frm 1985 t 1990, in ur cuntry, Vietnam, there were sme science prjects abut micrrganisms in TC1 fuel. At the present, as all cuntry in Asia, Vietnam used t utilize and imprt hundred tns f jet AI. Trpical climate f Vietnam is the suitable cnditin fr grwing f micrrganisms. With Cg - C I8 alkane chains, jet fuel is carbn surce fr a lt f micrrganisms, especially bacteria and fungi, which harmfully affect plane safety. Therefre, we aimed Vietnam airlines cmpany t cntrl the number f micrrganisms in jet fuel f all Vietnam airplanes, as well as study n the diversity f micrrganisms in trace water in fuel samples, lking fr the reasn why filter is bstructed t cntribute t the jet fuel string and safety in Vietnam. Materials and methds - Cllect JetAI samples frm tw water valves in tw airplane wings. - Inculate, islate and estimate the number f micrrganisms in selective media: R2A and MP A fr bacteria, Czapeck fr fungi, Hansen fr yeast, Gause fr actinmyces, Gst fr jet Al degrading bacteria. - Observe micrrganism cell mrphlgy then phtgraph micrrganisms under Japanese JEM 1010 electrmicrscpy. - Identify sme cmmn species in JetAl with bichemical tests f Bimerieux (API20NE, API50CHB, API 50CH) *Crrespnding authr. Tel.: ; fax: ; address: hien.pm@ibt.ac.vn -413-

3 - Study n the diversity f micrrganism ppulatin by 16S rdna analysis and DGGE methd. - Estimate the chemical and bilgical changes f jet fuel affected by micrrganisms with cntent methd and gas-chrmatgraphy. - Treat the data with Prs Nelsn Identify chemical and bilgical cmpnent f jet fuel filter in several planes t estimate the capacity f making bstructed filter f micrrganisms. Results and discussin The results invlved in three main parts: A. Analyzing the number f micrrganisms in trace water and fuel samples cllecting frm airplane at the time they land ff. B. Estimating the influence f micrrganisms n chemical cmpnent ffuel. C. Analyzing the micrrganisms and chemical structure f fuel filter samples. A. Analyzing the number f micrrganisms in trace water and fuel samples cllecting frm airplane at the time they land ff 1. The n.umber f micrrgan.isms in fuel trace water samples frm airplanes Table 1. The number f micrrganisms in fuel trace water samples frm airplanes f Vietnam airlines Sample n Sample The number f micrrganism (CFU/ml) Aerbic bacteria fungi JetAI degrader S7 -ASA 1.2 x 10 7 < S7 -ASA 7.6 x S7 -ASB 5 x 10 4 < 101 < S7 -ASC 1.5 x X S7 -ASD 4.7 x X S7 -ASD 10 6 < S7 -ASE 6.1 x S7 -ASE 2 x X S7 -ASF 6.6 x X S7 -ASF 1.2 x X S7 -ASG 2.4 x S7 -ASH x S7 -ASH 1.8 x 10 4 < S7 - ASI 8.4 x X S7 - ASI 2.4 x 10 5 < S7 - ASI 5.6 x 10 7 < A x A x A x X 10 1 < A x 10 6 < 101 < A x F x F x A x

4 In all 24 ftrace water samples in jet fuel micrrganisms are present. The minimum number f micrrganisms is 10 4 CFU/ml (S7-ASH n st.02) and the maximum number is 10 8 CFU/ml (S7-ASF n 5.26 th.03 with 10 8 bacteria/ml and 10 3 fungi/ml). Fungi are present in 19/24 analyzed samples and the maximum number f fungi is 6.6 x 10 5 CFU/ml in S7-ASD sample n 12.4 th.02. Only 5 samples have n fungi. JetAI degrading micrrganisms are present in samples. In sme samples, there are bth fungi and JetAl degrading bacteria (S7-ASD, 12.4 th.02; A , 1O.21st.03). Only in 3 samples, there are n JetAl utilizing micrrganisms. In an airplane but m different times f sample cllecting, the number f micrrganisms is different, depending n: the cllecting time, flight time and bicide treating time. 2. The number f micrrganisms in jet fuel samples Table 2. The micrrganism density in JetAl fuel frm planes f Vietnam airlines Density f micrrganism (CFU/ml) Day Samples Bacteria Fungi Yeast S7 -ASA S7 -ASA S7 - ASB S7 -ASC S7 -ASD S7 -ASD S7 -ASE S7 -ASE S7 -ASE S7 -ASF S7 -ASG S7 -ASH S7 -ASH S7 - ASI S7 - ASI S7 - ASI A A A A A F F A769 3 X X X x X X x X X X X X X X X X X X X X X X X X Actinmyces Jet Al degrader 10 1

5 Accrding t the table 1 and 2, micrrganisms in JetAl fuel in Vietnam are diverse with all representatin: aerbic bacteria, JetAl utilizing bacteria, yeast, actinmyces and fungi. Althugh cllecting sample at the same time and place, the number f micrrganisms in samples with trace water is thusand times higher than that in fuel samples. This means that it is necessary t cntrl and drain water in reservirs frequently, assciate string prcess with treating fuel filter during supplying plane. As a result, we will decrease the number f micrrganisms markedly. 3. DGGE analysis f bacterial ppulatin injetfuel taken frm airplanes After islatin crude DNA f ttal ppulatin in jet fuel we carried ut DGGE analysis (figure 1). There are many bands in sme clumn. The results f DGGE were very suitable t analyzed data n selective media (table 1 and 2). In the next step, we shuld cut these bands fr further analysis. Basing n mrphlgy and biphysical-bichemical characteristics, as well as bichemical tests and 16S rdna, we classified sme cmmn species in JetAl fuel B : ASA 3.4: ASB 5,6: ASI 7,8: A346 9,10: A769 Figure 1. DGGE f cntaminents in jet fuel Gram-negative bacteria invlved in Aermnas, Chrysemnas, Pseudmnas (p. aeruginsa, P. putida, P. jlurescens, P. cepacia) Gram-psitive bacteria invlved in Bacillus (E. cagulans; B. ciculans), Mycbacterium, Micrcccus Fungi invlved in Aspergillus (A. viridi, A.mutans, A. versiclol~ A. sydvy, A. jlavipes), Aurebasidium, Acremnium bacillispirilum, Paccillmyces varitii, Penicillium (p. lilacium, P. pullularia, P. nigricans, P. citrinum) Yeast invlved in Candida (c. trpicalis, C. albicans), Rhdturula (R. glutinis) Actinmyces invlved in Streptmyces (s. argentelus, S. rientalis, S. pulcher) Bacillus cagulans ASB Chrysemnas ATR- KOC Figure 2. The mrphlgy f Bacillus cagulans ASB and Chrysemnas ATR- KOC viewed with the electrmicrscpy -416

6 4. The number f micrrganisms in fuel samples frm reservir and filter Table 3. The rati f micrrganism grups in analyzed samples frm reservir and filter The number f The number f samples with micrrganisms micrrganism Aerbic Anaerbic JetAl degrading Actin- (CFU/ml) bacteria bacteria micrrganism myces Fungi Yeast > _ _ Ttal Table 4. The rati f fuel samples having micrrganism The number f The rati f fuel sample with micrrganisms (%) micrrganism Aerbic Anaerbic JetAl degrading Actinm (CFU/ml) bacteria bacteria micrrganism yces Fungi Yeast > ,5 0 1, _ ,3 8,3 21,5 19,2 11,3 15, _ ,1 41,6 27,2 30,7 38,0 35,0 50,0 50,0 49,4 50,0 50,0 50,0 Table 5. The number f micrrganism in jet fuel filter Samples Aerbic JetA1 degrading JetA1 degrading Actinbacteria bacteria fungi myces Fungi Yeast A320 3x10~ 2x ATR Accrding t the table 3 and 4, the number f micrrganism grups is in this rder: Aerbic bacteria> fungi> JetAl utilizing bacteria> actinmyces> yeast> anaerbic bacteria. All f these micrrganisms can take part in the JetAl fuel metablism. Sme strains are able t degrade fuel hydrcarbns; thers have capacity f utilizing mediatr prducts prduced by directly fuel utilizing bacteria. Therefre, the presence f all micrrganisms affect n quality f fuel, especially n ash weight after burning fuel. This amunt f ashes is mechanics mingling which makes the engine crrsive and shrten the life. We analyzed samples frm all airplanes f Vietnam airlines. Accrding t the data in table 6, we rely that the plane A320 and TU134 are the mst highly injected micrrganisms (56-57%). Particularly, it is very imprtant t pay attentin t JetAl utilizing aerbic bacteria and fungi, because they created viscus membrane in filter, s they make filter bstructed. Table 6. The prtin f infected micrrganisms in jet fuel Type f planes The number f The number f injected analyzed samples samples A Being Fkker ATR72 TUl Prtin (%) 56,0 22,7 20,0 23,3 57,0-417

7 B. Estimating the influence f micrrganisms n chemical cmpnent f fuel Table 7. The change f saturated hydrcarbn f JetA1 caused by micrrganisms Aurebasidium pullularia Pseudmnas sp. ASB Pseudmnas sp. ASJ TX1B1 The number Cntrl f carbn % % % utilized utilized utilized C7 0 C8 0 C9 67 CIO 1304 Cll 1939 C Cl C C C C17 46 C18 9 C19 trace Ttal 8036 Hydrcarbn remaining (mgl1) ,54 72,24 59,77 51,79 45,44 41,65 57,98 65,05 84,78 100,00 55,89 Hydrcarbn remaining (mg/l) ,54 17,79 9,24 5,20 9,08 26,07 44,66 56,52 17,47 Hydrcarbn remaining (mg/l) ,55 74,46 48,79 26,14 09,47 11,26 13,19 12,62 15,22 34, ,-----p'"\---;::::=============; Cntrl ~ --*f- Pseudmnas ASB E -;; \ Pseudmnas ASJ 'E 0 'm E ~ r~~~~~ c -e t1l e H-f-----~~~ ~ :t: k AU.pullularia TX1B1 C7 C8 C9 Cl0 Cll C12 C13 C14 C15 C16 C17 C18 C19 Number f carbns Fig.3. Degradatin f saturated hydrcarbn f JetAl by islated micrrganisms C. Analyzing the micrrganisms and cmpsitin f fuel filter samples Table 8. Analyzing the amunt f rganic cmpund in sediment f ASG plane filter Sample Percentage Sediment ffilter befre burning Sediment f filter after burning 0,0327 0,0124 The ammlt f rganic cmpund The amunt f inrganic cmpund 62,1 37,9 Accrding t the table 6, all 3 trains highly utilize saturated hydrcarbn f JetAl (frm 17.47% t 55.89%), especially CI0 - C17 hydrcarbn chains. Pseudmnas sp. ASB can degrade 100% C 18, Pseudmnas sp. ASJ degrades C15- C 17 hydrcarbn chain, and Aurebasidium pullularia TX1B1 degrades C9 - C 11 hydrcarbn chain. Bth bacteria and fungi change the JetAI hydrcarbn cmpnent

8 Organic cmpnent usually is frm 50 t 60% in sediment f filter, if this cmpnent is higher than 60%, this means that after burning, the amunt f rganic cmpund is created by micrrganisms. When the plane wrks this rganic cmpund will make filter bstructed. 5. The capacity f applying - We applied t cntrl the number f micrrganisms in jet fuel f Vietnam airlines - We can apply t all string units, petrleum and fuel Figure 4. Airplane filter string in Vietnam Cnclusin Frm analyzing mre than 200 jet fuel samples, we cncluded: 1. The number f micrrganisms in trace water jet fuel is frm 10 4 t 10 8 CFU/ml and. the number f micrrganisms is nly frm 10 1 t 10 3 in samples withut water. 2. Basing n mrphlgy and biphysical, bichemical characteristics, micrrganisms fund in Jet Al fuel are: Aermnas, Chlysemnas, Pseudmnas, Bacillus, Mycbacterium, Micrcccus, Candida, Rhdtrula, Aspergillus, Penicilium, Aurebasidium, Paecillmyces, Acremnium, Streptmyces 3. Accrding t the result f analyzing rganic and inrganic cmpund in filter, micrrganisms are the reasn that creates viscus membrane which make fuel bstructed. The main micrrganism structures n fuel filter are fungi and bacteria. 4. Analyzing hydrcarbn cmpnent f JetA1 under the effect f tw bacterial and ne fungus strain, the capacity f grwing and changing fuel cmpnent f micrrganisms brings abut the decrease the fuel quality. 5. It is necessary t cntrl the number f micrrganisms during fuel string frequently, drain water peridically and treat reservirs with bicide when the number f bacteria is higher than 10 2 CFU/ml. References Baryshnikva LM, Grishchenkv VG, Arinbasarv MU, Shkidchenk AN, Brnin AM., Bidegradatin f il prducts by degrader strains and their assciatins in liquid media] Prikl Bikhim Mikrbil. 37(5): Russian. Bent, F.M. and Gaylarde, C.C. Effect f additives n fuel stability - a micrbilgical study. In: LABS 3 - Third Latin American Bidegradatin and bideteriratin Sympsium, eds. C.C. Gaylarde, T.c. Barbsa and N.H.Gabilan, The British Phyclgical Sciety, UK, paper n. 10, Bent, F.M. and Gaylarde, C.c. Micrbial cntaminatin f stred diesel il in Brazil. Rev.Micrbil.,27: , Defene standard turbine fuel, aviatin kersene type JET AI. May 1996, DEF/STAN (DERD 2494)

9 Ferrari MD, Neirtti E, Albmz C "Occurrence f hetertrphic bacteria and fungi in an aviatin fuel handling system and its relatinship with fuel fuling" Rev Argent Micrbil Jul Sep; 30(3): Hill, E.C. and Hill, G.c. Micrbilgical prblems in distillate fuels. Trans. Inst. Marine Eng., 104: , lata guidance material fr aviatin turbine fuels. Aprils, Canada. Jnes RK, Sun WH, Tang CS, Rbert FM., Phytremediatin f petrleum hydrcarbns in trpical castal sils. ll. Micrbial respnse t plant rts and cntaminant. Envirn. Sci. Pllut. Res. Int. 11(5): Litman, E.S. Micrbilgical cntaminatin f fuels during strage. In Distillate Fuel Stability and Cleanliness, eds.l.l. Stavinah and E.I. Henry, ASTM , 1980, pp Saadun I., Islatin and characterizatin f bacteria frm crude petrleum il cntaminated sil and their ptential t degrade diesel fuel. 1. Basic Micrbil. 42(6): She1l1lan, J.L. Cntrl f micrbial cntaminatin f fuels in strage. In Bideteriratin 7, eds. DR Hughtn, R.N. Smith and H.O.W.Eggins, Elsevier, Barking, 1988, pp Slana, M.J.Y. and Gaylarde, c.c. Fungal cntaminatin and grwth in varius hydrcarbn fuels. In Bideteriratin and Bidegradatin 9, eds. E. Busher, M. Chandra and R. Edyvean, Rugby: Inst. Chern. Engs., 1995, pp Szbszlay S, Atzel B, Kriszt B., Cmparative bidegradatin examinatin f Pseudmnas aeruginsa (ATCC 27853) and ther il degraders n hydrcarbn cntaminated sil. 1: Cmmun Agric Appl BiI Sci. 68 (2 Pt A):207-1O. Whyte L.G., Hawari 1., Zhu I., Burb1l1liere L., l1l1liss W. E., Greer C. W., Bidegradatin f Variable-Chain-Length Alkanes at Lw Temperatures by a Psychrtrphic Rhdcccus sp. Appl Envirn. Micrbil., 64, 7: Yerushalmi L., Guit S.R., Kinetics f bidegradatin f gasline and its hydrcarbn cnstituents. Appl. Micrbil. Bitechnl., 49: