Annual Report 2010 - National Center for Genetic Engineering and ...

CONTENTSMessage from the BIOTEC Executive Director 5Facts and Figures 7Research and Development 15Commercialization and Private Sector Partnership 37Human Resource Development 45Public Awareness 53International Collaboration 57Impact of BIOTEC’s Output 65Appendices 71List of PublicationsList of Patents and Petty PatentsHonors and AwardsExecutives and Management Team

MESSAGE FROM THE BIOTECEXECUTIVE DIRECTOR2010 has proved to be a memorable year for BIOTEC and NSTDA. In June 2010, NSTDA bid farewellto Dr. Sakarindr Bhumiratana, who provided strong leadership as NSTDA President for the past 6 years.Dr. Sakarindr led BIOTEC as the Executive Director before taking up the office of NSTDA President. Underthe new leadership of Dr. Thaweesak Koanantakool, NSTDA will continue to position itself as a key partnerfor strengthening a knowledge-based society through science and technology.This year, a synthetic antifolate compound P218 targeting the dihydrofilate reductase enzyme of the parasiteP. falciparum and its drug resistant strains entered pre-clinical trials sponsored by Medicines for MalariaVenture (MMV). The design and synthesis of P218 are the results of research efforts from the teamat BIOTEC led by Prof. Yongyuth Yuthavong. This is the first time that a drug-like compound from anyThai research group was selected to enter preclinical safety testing. This has proven that Thailand has thecapability of moving beyond basic research phase to the next step in drug discovery.During the end of the rainy season of this year, Thailand faced an unexpected flood affecting vast areasfrom the Northeast to the central part of the country. Rice farms were severely damaged due to the rapidincrease in the high water level and the length of inundation by accumulated water due to poor drainage.BIOTEC has seen the positive results of a new strain of rice, Homcholasit with submergence tolerance,generated via the marker assisted breeding program headed by Dr. Theerayut Toojinda in collaborationwith the Rice Department and Kasetsart University, which was being tested in the field at the time of thefloods. This new rice strain can be completely submerged under water for approximately 20 days withoutlosing its vitality. Currently, more seeds are being distributed to farmers to propagate in preparation for nextyear’s planting season.As usual, throughout 2010 we welcomed researchers and students from different countries to be part of theBIOTEC research community. Besides enjoying the participation of researchers from the neighboring countriesunder our HRD Program for Asia Pacific, and returning students from Nanyang Polytechnic (Singapore),Temasek Polytechnic (Singapore), Atma Jaya Catholic University of Indonesia (Indonesia), BIOTEC alsowelcomed new comers from the University of Liverpool to spend 6 weeks as internees in BIOTEC labs.Located at Thailand Science Park, BIOTEC shares the park with private sector tenants providing endlessopportunity for our scientists to work side by side with companies. This year, we welcomed the openingof Virbac Animal Health-Biotechnology R&D Center in Asia located in BIOTEC’s incubator wing, and thelaunching of “KEEEN”, a bioremediation agent, developed by BIOTEC research team in collaboration withHi-Grimm Environmental and Research Co. Ltd. The product is a result of a two-year collaborative researchproject, starting in November 2008, between BIOTEC and Hi-Grimm on selecting oil-degrading bacteria foundin Thailand and developing culturing methods and media suitable for a variety of products.With our excellent team and strong alliances, BIOTEC hopes to advance science and technology to fulfill ourcommitment to building a stronger society and enhancing the competitiveness of the country.Dr. Kanyawim KirtikaraExecutive Director, BIOTEC

FACTS AND FIGURESBIOTEC was first set up under the Ministry of Science,Technology and Energy on 20 September 1983. Afterthe establishment of the National Science and TechnologyDevelopment Agency (NSTDA) on 30 December 1991,BIOTEC became one of the NSTDA centers, operatingoutside the normal framework of civil service and stateenterprises. This enabled the Center to operate moreeffectively to support and transfer technology for thedevelopment of industry, agriculture, natural resources,environment and consequently the social and economicwell-being of Thai people. Other centers under the NSTDAfamily include National Metal and Materials TechnologyCenter (MTEC), National Electronics and ComputerTechnology Center (NECTEC), National NanotechnologyCenter (NANOTEC) and Technology Management Center(TMC).As a premier research institute in Thailand and Asia,BIOTEC operates research units located at Thailand SciencePark and specialized laboratories hosted by variousuniversities, covering a wide spectrum of research topicsfrom agricultural science to biomedical science andenvironmental science. Apart from research laboratories,BIOTEC activities also include policy research, an outreachprogram, training and international relations.

8Annual Report 2010ORGANIZATIONBIOTECResearch Units• Research Unitslocated at ThailandScience Park• BIOTEC’s SatelliteUnits located atuniversities andother governmentdepartmentsBusiness Unit• Development Units• BusinessDevelopment andTechnologyTransferHRD and PlatformTechnologyManagement Division• Training Unit• PlatformTechnologyManagementAdministration• Planning Division• Evaluation &Monitoring Unit• Budgeting Section• Policy Study andBiosafety Unit• Rural DevelopmentTechnology ServiceUnit• Intelligence Unit• ManagementInformation Systems• Public RelationsSection• InternationalCooperation Division• OrganizationDevelopment Section• Facility ManagementSection• Procurement Section

National Center for Genetic Engineering and Biotechnology (BIOTEC) 9RESEARCH UNITSat Thailand Science ParkGenome InstituteBIOTECCentral Research UnitProtein-Ligand Engineering and MolecularBiology Research Program• Protein-Ligand Engineeringand Molecular Biology Laboratory• Virology and Cell TechnologyLaboratory• Anti-Tuberculous Drug ResearchLaboratoryAgricultural Biotechnology ResearchProgram• Food Biotechnology Laboratory• Monoclonal Antibody ProductionLaboratory• Animal Physiology Laboratory• Aquatic Molecular Genetics andBiotechnology Laboratory• Shrimp-Virus InteractionLaboratory• Microarray Laboratory• Starch Biosynthesis Laboratory• Plant Physiology & BiochemistryLaboratory• Plant Molecular GeneticsLaboratory• Plant Research GroupBioresourcesTechnology UnitBioresources Management Program• BIOTEC Culture CollectionLaboratory• Biotechnology LawDiscovery Program• Bioresources Research Laboratory• Bioassay Laboratory• Enzyme Technology Laboratory• Fermentation andBiochemical Engineering Laboratory• Microbial Engineering LaboratoryMicroorganism Program• Mycology Laboratory• Mushroom Cultivation ResearchLaboratory• Phylogenetics LaboratoryInformation System Program• Information Systems Laboratory• Geoinformatics Laboratory• Ecology Laboratory• Genomic Research Laboratory• Proteomics Laboratory• Biostatistics and InformaticsLaboratory

10Annual Report 2010RESEARCH UNITSat universities and government organization• Biochemical Engineering and Pilot Plant Research and Development Unit (BEC)- at King Mongkut’s University of Technology Thonburi (KMUTT)• Excellent Center of Waste Utilization and Management (ECoWaste)- at King Mongkut’s University of Technology Thonburi (KMUTT)• Cassava and Starch Technology Research Unit- at Kasetsart University• Rice Gene Discovery Unit- at Kasetsart University• Medical Biotechnology Research Unit- at Siriraj Hospital• Biomedical Technology Research Center- at Chiang Mai University• Center of Excellence for Marine Biotechnology- at Chulalongkorn University• Center of Excellence for Molecular Biology and Genomics of Shrimp- at Chulalongkorn University• Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp)- at Mahidol University• Peat Swamp and Rainforest Research Station- in Narathiwat ProvinceDEVELOPMENT UNITS• Shrimp Biotechnology Business Unit (SBBU)• Dairy Cattle Production Research and Business Development Project• Shrimp Genetic Improvement Center (SGIC)• Pilot Plant for Nuclear Polyhedrosis Virus (NPV) Production

National Center for Genetic Engineering and Biotechnology (BIOTEC) 11INCOME 1,111 million BahtEXPENDITURE 1,017 million Baht84 MBFunding and income earned from joint / contract researchand technical services405 MBTonkla Archeep Program (a government economy stimulusprogram)622 MBBudget allocated from NSTDA12 MB (1%)Technology Transfer36 MB (4%)Infrastructure Development and Maintenance123 MB (12%)Internal Management409 MB (40%)Human Resource Development437 MB (43%)Research and DevelopmentResearch and Development Expenditure 437 million Baht1 MB (0%)Underprivileged8 MB (2%)Energy42 MB (10%)R&D Management44 MB (10%)Platform technology55 MB (13%)Health and Medicine124 MB (28%)Environment163 MB (37%)Agriculture and Food

12Annual Report 2010HUMAN RESOURCES 544 employeesBIOTEC Staff by Job Function8 (1%)Executive127 (23%)Administrative409 (76%)R&D and Technical StaffBIOTEC Staff by Education32 (6%)Below B.S.155 (29%)PhD165 (30%)B.Sc.192 (35%)M.Sc.PUBLICATIONS 216 papers4In journals with impact factors more than 109In journals with impact factors between 4 and 1018In non-citation index journals185In journals with impact factors between 0 and 4

14Annual Report 2010

National Center for Genetic Engineering and Biotechnology (BIOTEC) 15RESEARCH ANDDEVELOPMENTBIOTEC’s R&D and technical program covers a widespectrum of research topics from agricultural tobiomedical sciences and from energy to environmentalscience. In addition to applied research, the Centeralso focuses on building up the nation’s capacity inplatform technologies.

16Annual Report 2010HIGHLIGHTS ON RICE BIOTECHNOLOGYImprovement of Rice Quality through Marker-assistedSelection Technology Rice has enormous economicvalue for Thai people. There are three ecosystems forrice production in Thailand including rain-fed lowland,irrigated lowland and upland. In rain-fed lowland, twofamous varieties namely RD6, a glutinous jasminerice and the non-glutinous jasmine rice Khao DawkMali 105 known as KDML105 are popularly grown byfarmers and are mainly intended for consumption andcommercial purposes. Although these varieties arehighly adapted to the rain-fed lowland environments,various constraints threaten their productivity bydecreasing yield and consequently affecting the livelihoodof farmers in the area. Problems in rice productioninclude damage from diseases and insects and stressfrom floods, drought and soil salinity.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 17The Rice Gene Discovery Unit, a specialized unitestablished by BIOTEC and Kasetsart University, haslong had research collaboration with the Bureau ofRice Research and Development to improve KDML105and RD6 against adverse conditions. Marker-assistedselection (MAS) was employed in developing lines tohasten the transfer of gene/QTL more efficiently.Three newly developed varieties are being field testedat the Rice Gene Discovery Unit in 2010. Theyare RD6 with 4 blast resistance genes pyramiding,non glutinous rice varieties with blight resistanceand non-photoperiod sensitivity, and non glutinousrice varieties with brown planthopper resistance andnon-photoperiod sensitivity. Three rice varieties;KDML105 with submergence tolerance and resistanceto blight and brown planthopper, KDML105 withtolerance to drought and salinity, and Surin 1 withdrought tolerance; are undergoing intensive fieldtrials at the Rice Department, where they are beingtested at various rice research stations and farmerfields across the country. One variety, KDML105with submergence tolerance, is being reviewed forvarietal registration. Two varieties, Homcholasit withsubmergence tolerance and non-photoperiod sensitivityand RD6 with blast resistance, are in the process ofapplying for varietal registration.In February 2010, NSTDA and the Rice Departmentsigned an MOU to extend R&D collaboration intothe next five years until 2015, aiming to developadditional varieties with desirable traits.

18Annual Report 2010HIGHLIGHTS ON PLANT BIOTECHNOLOGYRapid Regeneration of Oil Palm Cultivation of oilpalm has expanded enormously in recent years, dueto the surge in demand for cooking oil and biofuel.Since oil palm is a perennial crop species with a longbreeding cycle and a single growing apex, somaticembryogenesis has become the key method formultiplication of oil palm elite genotypes. In the studyconducted by the Genome Institute and KasetsartUniversity, researchers evaluated the effect of mediumcomposition and growth regulators on embryogeniccompetence and plant regeneration for the developmentof an efficient and rapid regeneration system viasomatic embryogenesis, aiming for commercial plantingof oil palm elite genotypes as well as genetictransformation studies. The study established a simpleand rapid procedure to improve commercial productionof clonal plantlets, using 13-week-old zygotic embryosas explants to culture on N6 medium with 2,4-D forcallus induction. Somatic embryos are produced onmaturation medium (N6 plus 2,4-D, puterscine, caseinamino acids and activated charcoal) for 3-5 months.In plant regeneration, modified N6 medium with

National Center for Genetic Engineering and Biotechnology (BIOTEC) 19activated charcoal was used without growth regulators,whereby both shoots and roots were inducedsimultaneously. This combined shoot and root inductionprotocol shortened the culture time to 9-12 monthsinstead of 18-24 months in normal protocol. Theprocedure will be extremely useful for commercialplanting of oil palm elite genotypes as well as genetictransformation studies. Results of this study werepublished in Acta Physiologiae Plantarum.Genome Sequence of Mungbean Mungbean, Vignaradiata, is an economically important crop grownprincipally for its protein-rich dry seeds. Mungbean iswidely used as an ingredient in Asian cuisine, in theform of seeds or bean sprouts. The extracted starchis used to make transparent cellophane noodles. Despiteits economic importance, genomic research ofmungbean has lagged behind other species in theFabaceae family. The research team at the GenomeInstitute, in collaboration with Kasetsart University,performed shotgun genome sequencing of V. radiatausing the 454 pyrosequencing technology in order toobtain more genomic information. 470,024 shotgunsequences were assembled to 46,646 contigs. Around1,500 microsatellites were identified that could be usedas potential DNA markers and 192 loci were validatedand demonstrated to be useful in Fabaceae. In addition,the researchers also observed that around 5% ofthe shotgun reads were derived from the mungbeanchloroplast genome. The complete chloroplast genomesequence was obtained and characterized. These twomajor works were reported in DNA Research andBMC Plant Biology.set of DNA markers and a genetic map. Researchersfrom the Genome Institute, Mahidol University and theDepartment of Agriculture developed additional simplesequence repeat (SSR) markers from the expressedsequence tags (ESTs), and constructed a geneticlinkage map. In the study, researchers designed 425EST-SSR markers from sequences obtained from thecassava EST database in GenBank, and integratedthem with 667 SSR markers from microsatelliteenrichedgenomic sequences received from theInternational Center for Tropical Agriculture (CIAT). Ofthese, 107 EST-SSR and 500 genomic SSR primerpairs showed polymorphic patterns when screened intwo cassava varieties, Huay Bong 60 and Hanatee,which were used as female and male parental lines,respectively. Within the 107 and 500 primer pairs,81 and 226 EST-SSR and SSR primer pairs weresuccessfully genotyped with 100 samples of F1 progeny,respectively. The results showed 20 linkage groupsconsisting of 211 markers—56 EST-SSR and 155SSR markers—spanning 1,178 cM, with an averagedistance between markers of 5.6 cM and about 11markers per linkage group. These novel EST-SSRmarkers provided genic PCR-based co-dominantmarkers that were useful, reliable and economical. TheEST-SSRs were used together with SSR markers toconstruct the cassava genetic linkage map which willbe useful for the identification of quantitative trait locicontrolling the traits of interest in cassava breedingprograms. This work was published in MolecularBreeding.Genetic Linkage Map of Cassava Use of moleculartechnology for genetic improvement of cassava (Manihotesculenta) has been limited by the lack of a large

20Annual Report 2010HIGHLIGHTS ON SHRIMP BIOTECHNOLOGYaddition, it is more sensitive than the traditional nestedRT-PCR. This first application of HRM multiplex RT-PCRprovides new potential for rapid and sensitivedetection of multiple pathogens in shrimp. The workwas published in Molecular and Cellular Probes.Successful Propagation of Shrimp Yellow Head Virusin Immortal Mosquito Cells Research on crustaceanviruses is hampered by the lack of continuous cell lines.A research team at Centex Shrimp partially solvedthe problem by challenging the C6/36 mosquito cellline with shrimp yellow head virus (YHV) and provedthat YHV can be propagated successfully in C6/36mosquito cells and could be used at low passagenumbers as a source of inoculum to initiate lethalinfections in shrimp. This leads to further studies thatrequire the cultivation of YHV. The results of this workwere reported in Diseases of Aquatic Organisms.Application of High Resolution Melt (HRM) Analysisfor Duplex Detection of Macrobrachium rosenbergiinodavirus (MrNV) and Extra Small Virus (XSV) inShrimp Rapid and sensitive detection of pathogens isthe key practice for prevention of disease outbreaksin shrimp farming. BIOTEC researchers at the Centerof Excellence for Shrimp Molecular Biology andBiotechnology (Centex Shrimp) combined a multiplexRT-PCR with high resolution melt (HRM) analysis for thesimultaneous detection of Macrobrachium rosenbergiinodavirus (MrNV) and extra small virus (XSV) infectionin the giant freshwater prawn, M. rosenbergii, as anexample for the application of HRM to detect pathogensin shrimp. The newly developed technique is probe-freeand does not require gel electrophoresis, thus speedingup the detection time and cost effectiveness. InA Clip Domain Serine Proteinase Plays a Role inAntibacterial Defense in Shrimp Shrimp lack anadaptive immune response but rely on an innateimmunity to protect themselves from surroundingpathogens. This defense system is able to eliminatethe invading pathogens efficiently after infection.Antimicrobial peptides (AMPs) are important componentsof the innate immune system and function as a firstline of defense against invading microorganisms bykilling or slowing the growth of microbes like bacteria,fungi, viruses and protozoa. BIOTEC researchers atthe Center of Excellence for Molecular Biology andGenomics of Shrimp identified the clip domain serineproteinases (Pmclip-SP1) in the black tiger shrimp(Penaeus monodon). This gene was up-regulated at3 hours and down-regulated at 6–48 hours followingVibrio harveyi infection. Suppression of the PmClipSP1gene by injection of double-stranded RNA (dsRNA)

National Center for Genetic Engineering and Biotechnology (BIOTEC) 21led to a significant increase in the number of viablebacteria in the hemolymph (approximately 2.4-fold)and in the mortality rate (59%) of V. harveyi-infectedshrimp. The findings suggest that PmClipSP1 playsa role in the antibacterial defense mechanism ofP. monodon shrimp. Results of this study were presentedin Developmental and Comparative Immunology.Molecular Characterization of Stress-relatedBiomarkers in the Giant Tiger Shrimp Penaeusmonodon Natural and farming environments can bevariable and unpredictable. Under intensive cultureconditions, shrimp can be exposed to suboptimalenvironmental conditions, such as elevated watertemperature, low dissolved oxygen and significantreductions in salinity levels causing significant stress.BIOTEC researchers isolated several stress-relatedgenes including calreticulin (CRT) and heat shockproteins (Hsp21, Hsp70 and Hsp90). Under the heattreatment (i.e. 35 ° C for 2.5 hours), the expressionlevels of Hsp21, Hsp70 and Hsp90 in hepatopancreasof juvenile shrimp were significantly increased for 3,2, and 11-fold. In gills, more prominent effects ofheat stress on Hsp90 were observed (23-fold in 2.5hours), while comparable increases on Hsp21 but noeffect on Hsp70 of heat shock were found. UnlikeHsp genes, CRT in gills and hepatopancreas aftertemperature stress were not significantly different, butthe expression profile of CRT in hemocytes wasup-regulated approximately 25 fold after treatmentat 35 ° C for 3 hours. Recombinant CRT protein wassuccessfully expressed in vitro and exhibited an abilityto form a complex with recombinant EndoplasmicReticulum protein 57 of P. monodon (rPmERp57). Theinformation indicated that Hsps and CRT genes canbe regarded as biomarkers for temperature stressresponses in P. monodon. This work was publishedin Developmental and Comparative Immunology.Molecular Cloning and Expression of Progestinmembrane receptor component 1 (Pgmrc1) andProgesterone receptor-related protein p23 (Pm-p23)of the Black Tiger Shrimp Penaeus monodonGenetic improvement of P. monodon cannot beachieved without knowledge on the control of itsreproductive maturation. However, reduced spawningpotential and low degree of maturation of P. monodonin captivity crucially prohibit the development of effectivedomestication and selective breeding programs in thisspecies. Knowledge on molecular mechanisms of steroidhormonal induction on oocyte development maylead to possible ways to effectively induce ovarianmaturation in shrimp. BIOTEC researchers successfullyidentified PmPgmrc1 and Pm-p23 of P. monodon.These reproduction-related genes were differentiallyexpressed during ovarian development of P. monodon.Recombinant PmPgmrc1 and Pm-p23 proteins andtheir polyclonal antibody were produced and theirexpression analysis suggested that these geneproducts seem to play an important role in ovariandevelopment and may be used as a bioindicator formonitoring the progression of oocyte maturation ofP. monodon. This work was published in General andComparative Endocrinology.

22Annual Report 2010HIGHLIGHTS ON FOOD SCIENCE, TECHNOLOGYAND BIOTECHNOLOGYthe Ministry of Agriculture and Cooperatives. Thestudy concluded that applying the current standardof 200 ppm histamine concentration or alternativelya standard of 400 ppm does not greatly affect riskto consumers. The result of this study was used bythe Delegation of Thailand to propose to the 30thSession of the Codex Committee on Fish and FisheryProducts (CCFFP) taking place on 24 September – 4October 2009 in Morocco for the adjustment in thehistamine concentration in fish sauce to 400 ppm (40mg histamine /100g of fish sauce). The proposal wasendorsed by the Committee. This action can help theThai fish sauce industry to overcome trade barrierspreviously caused by conservative standards.Utilizing Microorganisms to Degrade Histamine inFish Sauce As high levels of histamine pose a riskof food toxification, BIOTEC Food BiotechnologyLaboratory in collaboration with Prince of SongklaUniversity and Chulalongkorn University, developed theRisk Assessment of Histamine in Fish Sauce Thepresence of high levels of histamine is detrimentalto the quality and safety of food, particularly fishsauce. Since fish sauce is a staple seasoning ingredientin Thai cuisine, with a value of 6.6 billion Baht fordomestic consumption and 1 billion Baht in exports,a joint study to assess the risk of histamine in fishsauce was launched at BIOTEC Food BiotechnologyLaboratory, in collaboration with the Department ofFisheries and The National Bureau of AgriculturalCommodity and Food Standards (ACFS), both under

National Center for Genetic Engineering and Biotechnology (BIOTEC) 23and temperature. This work was reported in FoodChemistry and could be applied directly in fish sauce.From these studies, a patent on “Biological processfor degradation of histamine in high salt containingfoods by using halophilic archaea producing histaminedehydrogenase” was filed in November 2009.use of halophilic archaea to degrade histamine in highsalt-fermented fishery products, including fish sauce.In 2008, the team reported on a novel halophilicarchaeon, namely Natrinema gari BCC 24369 thatwas isolated from salted anchovy. Later the study,published this year in Enzyme and Microbial Technology,revealed that this selected strain is capable of reducinghistamine in high salt condition and its histaminedegradingactivity is mediated through the intracellularhistamine dehydrogenase. Due to the limitations ofapplying Nmn. gari BCC 24369 cells and its enzymecaused by the slow growth rate, low yield of enzyme,and the cost of cell or enzyme preparation, a studyon the immobilization of whole cells to providepotential advantages over applications of free cellsand enzyme systems was performed. The immobilizedwhole cells retained histamine-degrading activity ashigh as 94% of the original activity detected in freewhole cells. Immobilized whole cells rendered stablehistamine-degrading activity at high salt concentrationScreening for Starter Culture for Fermented PorkSausage Nham is a Thai traditional fermented porksausage made from minced pork, boiled pork rinds,cooked rice, garlic, salt, sugar, pepper, chili and sodiumnitrite, packed in banana leaves or plastic sheetsand allowed to ferment for approximately 3–4 days.Over-fermentation of Nham, resulting in a variety ofundesired flavor and texture changes such as waterdripping, discoloration and off-flavor development, cantake place during handling, transportation or storageif not refrigerated. There is considerable interest inan economic solution to prevent over-fermentation.A mutated starter culture whose growth is sensitiveto high acidity, i.e. pH 4.6, the desirable pH of Nhamspecified by Thailand Food Standards and applythem in Nham fermentation, can offer a strategy toovercome the over-fermentation problem.In the study conducted jointly by BIOTEC FoodBiotechnology Laboratory, Mahidol University and theCenter of Excellence for Agricultural Biotechnology,several acid-sensitive mutants of one of the commercialNham starter cultures, Lactobacillus plantarum BCC9546, were isolated as spontaneous neomycin-resistantmutants. The most acid-sensitive mutant was appliedin experimental Nham production and the pH of Nhamfermented with the mutant was significantly higherat the end of fermentation (3 days) and after anadditional 4 days of storage at 30 ° C. These resultsindicate that the use of acid-sensitive L. plantarumas starter culture can reduce the severity of postacidificationand increase the shelf life of Nham atambient temperature. The results of this study werepublished in Food Microbiology.

24Annual Report 2010HIGHLIGHTS ON BIORESOURCES STUDYAND UTILIZATIONMicrobial Utilization in Animal Feed Industry BIOTECCulture Collection (BCC) holds more than 40,000strains of microorganisms, consisting of 68%filamentous fungi, 23% bacteria, 8% yeast and 1%algae. Part of the work conducted by the BioresourcesTechnology Unit is to find application for these localbioresources in the animal feed industry. Four enzymes,namely mannanase, phytase, xylanase and cellulaseproduced from in-house microbial collection are beinginvestigated. Mannanase enzyme is being produced atpre-pilot scale for testing in a chicken farm, to studythe potential use as a feed supplement. Thermostablerecombinant phytase has been tested to be on par withcommercially available phytase. Scale-up productionof 200 L is being conducted, while the pre-pilot scalelevel (50L) has already been achieved. Xylanase and

National Center for Genetic Engineering and Biotechnology (BIOTEC) 25cellulase from Thai-isolated fungi were tested and wereshown to improve fiber digestion and consequently,to increase metabolizable energy in poultry. Testingwith pigs is currently being planned with a commercialfarm. Apart from work with enzymes, BIOTEC scientistsalso developed high-density cultivation technology forSchizochytrium production in a 300 L fermentor. Withthis technology, up to 24 g/l of DHA is obtained. Thetechnology will likely be of interest to the aquaculturefeed industry.Enzyme Discovery for Ethanol Production fromBagasse Bagasse, the solid residue left after extractionof sugarcane juice, is one of the major lignocellulosicplant residues. With its high cellulosic polysaccharidecontent, it is considered as a potential source forethanol production. Researchers from the BioresourcesTechnology Unit conducted experiments on enzymatichydrolysis and fermentation of pre-treated bagasse,using enzyme-producing fungi and yeast strainsavailable in the BIOTEC Culture Collection. Theenzymatic hydrolysis stage utilizes an enzyme cocktailmade up of crude enzymes prepared from Penicilliumchrysogenum BCC4504 (containing cellulase activity) andAspergillus flavus BCC7179 (containing complementaryβ-glucosidase activity); whilst the fermentation stepuses Pichia stipitis. This study demonstrated thepotential use of local microorganisms in the productionof ethanol from this potent lignocellulosic biomass.The lab scale results of the experiment have beenreported in the Journal of Bioscience and Bioengineering.The research team also developed a multi-enzyme,consisting of non-starch polysaccharide hydrolyzingenzyme and raw starch degrading amylolytic enzyme,prepared from local Aspergillus in the BIOTEC CultureCollection for cassava hydrolysis. These multi-enzymescan be utilized with various forms of cassava, suchas fresh roots, chips and pulp in the non-thermalhydrolysis and saccharification process. Studies onthese multi-enzymes are underway for scale-upproduction and its effectiveness in various applicationsincluding biogas production and as an animal feedsupplement.Biocontrol Application To search for potentialbiocontrol agents, isolates maintained at BCC havebeen screened for their toxicity against the economicallyimportant insect pests. Beauveria bassiana BCC 2660was evaluated for biological control of green peachaphid, Myzus persicae (Sulzer), in kale greenhouse. Theperformance is 4 times better than the commercialformulation. Currently, scientists are working incollaboration with the Rice Department and theDepartment of Agriculture to test its effectiveness incontrolling brown plant hopper (Nilaparvata lugens)and Myzus persicae in the field. In the Bacillusthurigiensis study, scientists discovered the Vip3Aaprotein to be highly effective on beet armyworm(Spodoptera exigua) and leafworm (Spodoptera litura),when compared to protein extracted from commercialBt. Field testing of Vip3Aa protein is being plannedwith TFI Green Biotech Co., Ltd., a manufacturer ofBt for biocontrol.

26Annual Report 2010HIGHLIGHTS ON MALARIA RESEARCHDevelopment of Effective Antifolate Drug Fordecades, the group at BIOTEC Protein-LigandEngineering and Molecular Biology Laboratory havelong been engaged in research towards developingantimalarial drugs to overcome multi-drug resistantmalaria. The main strategies revolve around rational drugdesign and the synthesis of new effective antimalarialsbased on the structures of the drug targets. The team

National Center for Genetic Engineering and Biotechnology (BIOTEC) 27properties after oral administration in rodents and wasremarkably efficacious against Plasmodium berghei,leading to fully cured infected mice. In the course ofthese efficacy studies, however, some dose limitingtoxicity was uncovered at higher doses. In conclusion,despite its relative in vitro and in vivo selectivitytoward the Plasmodium DHFR enzyme and malariaparasites, QN254 does not show the adequatetherapeutic index to justify its further development asa single agent. This work was reported in AntimicrobialAgents and Chemotherapy.has discovered the protein structure of dihydrofolatereductase-thymidylate synthase of Plasmodium falciparum(PfDHFR-TS), an important drug target of Plasmodiumparasites. Based on information obtained from co-crystalstructure of DHFR and selected compounds, researcherswere able to design and create better effectivecompounds against resistant parasites. Initial safetytesting indicated that one of these compounds, P218,has a good safety margin between the toxicity inanimals and the predicted effective human dose. Thecompound will be examined by another group for itsparasite reduction ratio (PRR) value and propensity togenerate new resistance before further decision by theMedicines for Malaria Venture (MMV).Preclinical Evaluation of Antifolate QN254 Researchersin BIOTEC Protein-Ligand Engineering and MolecularBiology Laboratory took part in a preclinical evaluationof antifolate QN254 launched by Novartis Institutefor Tropical Diseases and Wellcome Trust. BIOTECresearchers performed the inhibitory activity test ofQN254 against pfDHFR and determined the crystalstructure of compounds in PfDHFR-TS. The teamprovided biochemical and structural evidence thatQN254 binds and inhibits the function of both thewild-type and the quadruple-mutant (V1S) forms ofthe DHFR enzyme. Study by other partners in theproject found that the compound is highly activeagainst Plasmodium falciparum clinical isolates,displaying various levels of antifolate drug sensitivity.The compound displayed favorable pharmacokinetic

28Annual Report 2010HIGHLIGHTS ON DENGUE RESEARCHfrom the Medical Biotechnology Research Unit, a jointunit between BIOTEC and the Faculty of MedicineSiriraj Hospital, and Emory University, it was for thefirst time found that rhesus macaques inoculatedintravenously with a high dose of dengue virus serotype2 (strain 16681) produced dengue hemorrhage, oneof the key clinical manifestations detected in denguepatients. The established monkey model may thereforeprovide a unique platform to define the early eventsin dengue virus infection and help identify which bloodcomponents contribute to the pathogenesis of denguedisease. This work was published in Blood.Role of Innate Immune Responses in DengueVirus Infection Role of innate immune parameters inthe course of dengue virus infection and pathogenesisof dengue is poorly understood. The main dilemma isdue to the lack of satisfactory small animal models fordengue virus infection. The only animal species besideshumans that are known to be naturally infected andthat can be experimentally infected via the parenteralroute are monkeys and apes. Previous studies usingsubcutaneous and/or intramuscular experimentalinoculation of rhesus macaques with dengue virusdemonstrated the antibody and viremia response inthe pattern relatively similar to that in man. However,clinical sequelae typical of human dengue virus infectionhave never been observed in the previously existingmonkey models. In a joint study between scientistsA Re-evaluation of the Mechanisms Leading toDengue Hemorrhagic Fever Viremia is one of theimportant features of dengue virus infection among theflaviviruses. Dengue virus infection results in a spectrumof clinical symptoms, ranging from undifferentiatedflu-like illness, mild dengue fever, to dengue hemorrhagicfever (DHF)/dengue shock syndrome (DSS), alife-threatening illness. Several mechanisms have beenhypothesized based primarily on data collected frompost-acute clinical phase to account for DHF/DSS.Among all the hypothesized mechanisms that havebeen forwarded to date concerning the etiology ofDHF, the understanding of the dynamics of dengueviremia is perhaps the most critical since this canpotentially influence vaccine design to prevent denguevirus infection and dengue disease development. Scientistsfrom the Medical Biotechnology Research Unit andEmory University presented a novel alternative conceptregarding how dengue virus disseminates during theacute viremia period based on its intimate relationshipwith platelets. They demonstrated that dengue viralparticles are present within platelets isolated fromdengue patients and that dengue viral products couldbe detected within platelets experimentally infected

National Center for Genetic Engineering and Biotechnology (BIOTEC) 29in vitro. Their results suggested that platelets maybe involved in the initial replication and disseminationof dengue virus in infected humans and may shielddengue virus from the host immune system. Thiswork, therefore, provides a useful piece of informationregarding the status of dengue virus and its potentialsurvival strategy in circulation during acute infection.The study was reported in Annals of the New YorkAcademy of Sciences.Cross-Reacting Antibodies Enhance Dengue VirusInfection in Humans In a collaborative researchbetween Imperial College London, Faculty of MedicineSiriraj Hospital Mahidol University, Khon Kaen Hospital,Songkhla Hospital, and the Medical Biotechnology Unit,scientists have analysed a panel of dengue specifichuman monoclonal antibodies (hAb) from human Bcells isolated from dengue infected patients. Antibodiesto the structural precursor-membrane protein (prM) forma major component of the response. These antibodiesare highly cross-reactive among the dengue virusserotypes and, even at high concentrations, do notneutralize infection but potently promote antibodydependent enhancement (ADE). ADE is thephenomena whereby non-neutralised serotypecross-reactive antibodies amplify the number of virusinfected cells by binding to virions as immune complexwhich then be taken up into Fc receptor bearingcells, leading to more infected cells. ADE has beenproposed as being one of the factors contributing tosevere dengue disease (Dengue Hemorrhagic fever,DHF). Anti-prM hAbs can enhance virus infection innot only cell lines but also in primary cells such asmonocytes and dendritic cells. Scientists propose that thepartial cleavage of prM from the viral surface reducesthe density of antigen available for viral neutralization,leaving dengue viruses susceptible to ADE by antibodyto prM. This finding has implications for future vaccinedesign in which the dengue vaccine should not induceanti-prM Ab responses in order to avoid ADE. Thework was published in Science.Immunodominant T-cell Responses to DengueVirus NS3 Are Associated with DHF In this study,scientists set out to study T-cell responses across theentire dengue virus proteome and to see whetherthese were related to disease severity in a cohortof dengue-infected children from Thailand. Robustresponses were observed in most infected individualsagainst most viral proteins. Responses to denguenon-structural protein 3 (NS3) were the most frequent,and there was a very strong association betweenthe magnitude of the response and disease severity.Furthermore, in severe dengue cases (DHF cases)cytokine-high CD107a-negative cells predominated,suggesting that these cells may play a role inimmunopathogenesis of the dengue disease. Thiswork was published in the Proceedings of the NationalAcademy of Sciences (PNAS).Influence of pr-M Cleavage on the Heterogeneityof Extracellular Dengue Virus Particles During Flavivirusreplication, new virus progenies containing envelope (E)and prM glycoproteins on their surfaces are generatedas non-infectious, immature virions. Cleavage of theprM proteins by host furin-like protease releasesextracellular infectious, mature virions, containing E andembedded M proteins, from infected cells. However,an incomplete cleavage of the prM glycoprotein of denguevirus generates a mixture of mature (prM-less) andprM-containing, immature extracellular particles. In thecollaborative research between Chiang Mai University,Purdue University, Faculty of Medicine Siriraj Hospitaland BIOTEC, “Partially mature virion”, as a third typeof extracellular particles, the major prM-containingparticles in a dengue serotype 2 virus, was demonstratedby sequential immunoprecipitation and cryo-electronmicroscopy. Changes in the proportion of viral particlesin the pr-M junction mutants exhibiting altered levelsof prM cleavage suggest that the partially matureparticles may represent an intermediate subpopulationin the virus maturation pathway. These findings areconsistent with a model suggesting the progressivemode of prM cleavage. The results of this studywere presented in the Journal of Virology.

30Annual Report 2010HIGHLIGHTS ON INFLUENZA RESEARCHExtraction of Neuraminidase of H5N1 Influenza VirusUsing Thrombin Proteolytics BIOTEC researchersemployed reverse genetics to construct non-pathogenicrecombinant influenza A viruses, termed rgH1N1 LVPRand rgH1N1 LVPR-GS, that harbor the neuraminidase(NA) of H5N1 virus engineered to contain a specificthrombin cleavage site at the stalk region. By usingthrombin to cleave NA at its stalk, a productiveextraction of NA globular heads could be obtainedfrom purified rgH1N1 LVPR. Furthermore, it was foundthat the NA of rgH1N1 LVPR-GScould be cleaved byendogenous thrombin present in embryonated chickeneggs, resulting in the release of NA globular heads intoallantoic fluids. These data highlight the use ofthrombin cleavage as an effective strategy for extractionof active NA heads directly from live viral particlesnot only of H5N1 but, theoretically, of any subtype ofinfluenza A viruses. This work was published in theJournal of Virological Methods.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 31Prediction of Avian Influenza A Binding Preferenceto Human Receptor It is known that the highlypathogenic avian influenza A virus H5N1 binds stronglyand with high specificity to the avian-type receptorby its hemagglutinin surface protein. This specificity isnormally a barrier to viral transmission from birds tohumans. However, strains may emerge with mutatedhemagglutinin, potentially changing the receptor bindingpreference from avian to human-type. The changein binding preference is due to mutation, which canbe computationally modelled. BIOTEC researchers incollaboration with Kasetsart University, used theinformation from the available structures of HAcomplexed with avian and human receptors to predictthe HA binding selectivity from different HA variants inreceptor-based conformational analysis by measuringa single torsion angle during molecular dynamicsimulation and found the predictions of receptorpreference agreeable with the available in vitro bindingdata. This work was published in BMC Genomics.

32Annual Report 2010HIGHLIGHTS ON BIOINFORMATICSAND INFORMATION SYSTEMSHuman Genetic Mapping of Asia BIOTEC researcherswere among 90 scientists participating in the HumanGenome Organisation’s (HUGO’s) Pan-Asian SNPConsortium, engaging in the study to map geneticdiversity in Asia, covering 73 Southeast Asian (SEA)and East Asian (EA) populations. This human geneticmapping of Asia has important implications, especiallyin the further understanding of migratory patterns inhuman history, and for the study of genetics anddiseases. The findings were published online in a reportin Science on 10 December 2009. In a genome-wideanalysis of genetic variation using more than fiftythousand single nucleotide polymorphisms (SNPs)typed in thirteen population samples from Thailand,it was found that Mlabri, a group of nomadichunter-gatherers inhabiting the rural highlands ofThailand, share more recent common ancestry withthe Htin. This can potentially be the first geneticevidence that supports the linguistic affinity of Mlabri, andthis association between linguistic and geneticclassifications could reflect the same past populationprocesses. The study was published in BMC Genetics.RExPrimer: An Integrated Primer Designing ToolScientists at the Genome Institute have developed anintegrated graphical web-based application for primer

National Center for Genetic Engineering and Biotechnology (BIOTEC) 33design, called RExPrimer, which was written in Pythonlanguage. RExPrimer provides many functionalitiesfor improved PCR primer design. Several databases,namely annotated human SNP databases, insertion/deletion (indel) polymorphisms database, pseudogenedatabase, and structural genomic variation databaseswere integrated into RExPrimer, enabling an effectivewithout-leaving-the-website validation of the resultingprimers. By incorporating these databases, the primersreported by RExPrimer avoid mis-priming to relatedsequences as well as possible PCR failure because ofstructural polymorphisms. In the study published inBMC Genomics, researchers demonstrated the programeffectiveness in successfully generating primers forstrong homologous sequences. This software is freelyavailable at http://www4a.biotec.or.th/rexprimer.ipPCA: A Novel Population Structure AnalysisAlgorithm Population structure analysis is importantto genetic association studies and evolutionaryinvestigations. As genotypic datasets becomeever larger, population structure analysis becomesprogressively difficult. Therefore, to keep abreast withthe ever increasing size and complexity of genotypicdata, BIOTEC researchers, in collaboration withresearchers from NECTEC and the Faculty of MedicineSiriraj Hospital, developed a novel population structureanalysis algorithm called iterative pruning PCA (ipPCA).In the study reported in BMC Bioinformatics, ipPCAperformance was tested on real datasets and able toaccurately resolve even highly structured populationscontaining many closely related subpopulations. Thissoftware is freely available at http://www4a.biotec.or.th/GI/tools/ippca.iCollect: An Enhanced Solution for BiologicalResources Management iCollect is a software tosupport laboratory inventory management of biologicalcollections at BIOTEC. iCollect is designed with ahighly customizable, configurable and user-friendlyinterface to allow users to create and store samples;graphically view the content of samples and searchinformation in a collection. Using the barcode assignedby iCollect, users can easily find a sample’s positionin the storage, and add a sample to a selectedstorage position. The in and out movement ofsamples in the storage are recorded and tracked foraudit trail. Moreover, the graphical tree view of thesamples can be used to trace the aliquot/sample backto its original parent.

34Annual Report 2010HIGHLIGHTS ON RISK ASSESSMENTAND BIOSAFETYBiosafety Guidelines for Contained Use of GeneticallyModified Microorganisms (GMM) at Pilot andIndustrial Scale BIOTEC, as the Secretariat to theTechnical Biosafety Committee (TBC), released theBiosafety Guidelines for Contained Use of GeneticallyModified Microorganisms (GMM) at Pilot and IndustrialScale in November, 2010. The Guidelines provide animportant framework for researchers and entrepreneursto utilize genetically modified microorganisms in allindustrial production such as food industries, agricultureindustries and environmental industries etc. Thedocument outlines the safe handling of GMM in pilotand industrial scale, covering the subjects of thecontainment and facility management to protect itsexposure to workers or environment.OECD Consensus Document on CompositionalConsiderations for New Varieties of Papaya Since1999, OECD’s Task Force for the Safety of NovelFoods and Feeds has focused its work on thedevelopment of science-based consensus documents,which are mutually acceptable among membercountries. These consensus documents containinformation for use during the regulatory assessment

National Center for Genetic Engineering and Biotechnology (BIOTEC) 35material on the production, processing and uses ofpapaya and considerations to be taken into accountwhen assessing new varieties of papaya. Constituentsto be analysed, related to food use and to feed use,are suggested. The Document was endorsed by TheTask Force and released for public access on theOECD website in June 2010. Among plants and cropsincluded in the previous consensus documents werecanola, soybean, sugar beet and potato. Thailand isthe first non-OECD member country to lead thedevelopment of a consensus document.of a particular food/feed product. In the area offood and feed safety, consensus documents are beingpublished on the nutrients, anti-nutrients or toxicants,information of its use as a food/feed and other relevantinformation. Thailand took the lead, in collaboration withthe United States (co-lead), to develop the consensusdocument addressing compositional considerations fornew varieties of papaya (Carica papaya L.) by identifyingthe key food and feed nutrients, anti-nutrients, toxicantsand allergens. A general description of thesecomponents is provided. As well, there is background

COMMERCIALIZATION ANDPRIVATE SECTORPARTNERSHIPBIOTEC places strong emphasis on promoting theindustrial applications of biotechnology in both Thaiand foreign companies. Technology/product licensingas well as collaborative research with the privatesector are among mechanisms to promote bio-businessin Thailand. Recognizing that the majority of companiesin Thailand are of small and medium size and do nothave in-house R&D capability, BIOTEC also providesR&D services in the form of contract research.

38Annual Report 2010LICENSING AGREEMENTSDuring FY 2010, six licensing agreements wereaccomplished with the following clients.SHRIMP BIOTECHNOLOGY BUSINESS UNIT (SBBU)SBBU was granted the rights to manufacture anddistribute Taura Syndrome Virus (TSV) and WhiteSpot Syndrome Virus (WSSV) detection kits basedon the LAMP-LFD technique. Compared to PCR-basedmethods, these test kits are economical, swift andsimple to use.MBSAsia Ltd. The licensing agreement covers thepermission to distribute monoclonal antibodies forprogesterone hormones. Progesterone levels can beused to confirm that cows are in estrus phase beforeperforming artificial insemination, as well as to confirmtheir pregnancy.Innova Biotechnology Co., Ltd. Under the agreement,Innova Biotechnology has the rights to produce anddistribute detection kits for white leaf disease insugarcane. These kits can be used on-site to detectphytoplasma directly in molasses. They can be usedas a risk management tool to reduce the spread ofand damage from the white leaf disease.New World Biotech Co., Ltd. New World Biotechacquired the rights to manufacture and distribute

National Center for Genetic Engineering and Biotechnology (BIOTEC) 39Infectious Myonecrosis Virus (IMNV) detection kitsbased on the LAMP-LFD technique. They can beused to reduce and contain IMNV infection in shrimpaquaculture.TFI Green Biotech Co., Ltd. TFI Green Biotech wasgranted the production rights on Bacillus megateriumfor the purpose of evaluating its efficacy againstRhizoctonia solani, which causes rice sheath blight. B.megaterium has been formulated into stable, effective,easy-to-use and safe biopesticide.Drew-Bio (Thailand) Co., Ltd. Drew-Bio (Thailand)obtained the rights to evaluate the performanceof prototypical tools that can interpret types ofThalassemia, Thalassemia carriers, and abnormalhemoglobin from hemoglobin type result obtained fromLow Pressure Liquid Chromatography (LPLC). Theseprototypes, together with mean corpuscular volume(MCV) information, can be added-on to any LPLCfor automatic Thalassemia test result interpretation.This system uses computer and electronic technologiesto intercept the print out signed from LPLC andanalyze it together with MCV data to offer theThalassemia type interpretation for physicians andmedical technicians.

40Annual Report 2010LAB TO MARKET HIGHLIGHTSSeveral products and prototypes developed fromBIOTEC laboratories have been introduced this yearand some examples are:iBrew Bovine Meat Quality Genotyping Kit iBrewcan be used to predict the meat quality of beefcattle. The Kit works by determining the genotypesof four Single Nucleotide Polymorphisms (SNPs) inCalpastatin (CAST), Calpain (CAPN) and Thyroglobulin(TG) genes that influence meat quality traits, interms of tenderness and juiciness, in cattle by usingthe restriction fragment length polymorphism (RFLP)method. iBrew uses non-toxic chemicals and is alsouser-friendly, giving results within 3 hours. The testwill be useful in making the selection of sires anddams for breeding, as well as predicting the meatquality of calves and cattle.Azupure Sperm DNA Purification Kit Azupure is agenomic DNA kit for extraction and purification fromfresh or frozen semen and whole blood of mammalssuch as humans, canines and cattle. Employing asimplified and rapid centrifugation method for DNAextraction process, the Kit offers short preparation timesand fast extraction of sperm DNA in approximately2 hours, using non toxic chemical reagents. Azupurecan be applied in forensic science, mainly in theexamination of semen stains; medical science such asexamining male infertility; as well as livestock breedingand improvement. It is suitable for on-farm use, aswell as in a research laboratory.ENZstrips ENZstrips are test strips containing in-gelcrosslink substrate for detecting cellulases, hemicellulases,beta-glucanases and amylases enzyme activity for

National Center for Genetic Engineering and Biotechnology (BIOTEC) 41Aqua-RAS D Aqua-RAS D is a closed recirculatingaquaculture system integrated with hybrid nitrificationbiofilter tanks for ammonia removal and patentedTubular Denitrification Reactor (TDNR) for nitrateremoval. With TDNR, water can be recirculated withinthe system hence water exchange can be prolongedfor a year or more.K9 Diagnostic Kit-MDR1 K9 Diagnostic Kit-MDR1 isa canine drug allergy test kit, using blood samples.The results can be obtained within an hour, making itsuitable for animal clinics and hospitals, as well as foranimal owners to perform the test at home.industrial and academic research use. To detectenzyme activities in the sample, the test strip, madeof plastic polymer, is dipped in the sample solution orextract. Enzymes in the sample will specifically attackits chromogenic substrates immobilized in the gelattached to the strip. Reactions can be incubatedunder a wide range of conditions and would takefrom a few minutes to hours depending on enzymeactivity level in samples. Once color is observed, itmeans that enzyme activity has taken place.This year, Hi-Grimm Environmental and Research Co.Ltd., launched a bioremediation product line under thebrandname “KEEEN”. KEEEN is the result of atwo-year collaborative research project, betweenBIOTEC and Hi-Grimm Environmental and ResearchCo. Ltd., for selecting oil-degrading bacteria found inThailand and developing culturing methods and mediasuitable for a variety of products. KEEEN providesits services into seven lines of business to caterfor the specific demands of its customers; 1.Solution Wastewater Treatment, 2. Solution Oil TankDegassing/Cleaning Remediation, 3. Solution Oil SpillResponse/Clean Up, 4. Solution Industrial Application,5. Solution Site Contamination and RemediationService, 6. Solution Sanitary Treatment and 7.Solution Testing/Diagnosis/Research.BIOTEC researchers are encouraged to participate invarious activities implemented by NSTDA designedto support their “Lab to Market” strategy, such asbootcamps, early-stage technology commercializationplan competitions (I2P competition) and technologybusiness plan competitions (Technopreneur competition).Through this incubation, researchers gain criticalfeedback and advice about the business model,management team, competitive positioning, marketstrategy and other key elements to adjust theirtechnology to better address market opportunities. Asa result of these programs, the following two BIOTECinventions were featured at NSTDA Investors’ Day,a public event to introduce NSTDA’s products andinventions to potential investors:

42Annual Report 2010COLLABORATIVE RESEARCHPTT Chemical Public Company Limited (PTTCH)Development of gamma-linolenic acid production fromMucor circinelloidesArysta LifeScience Corporation (Japan)Rearing and production of granulosis virus tocontrol leaf roller caterpillarsBIOTEC conducted a total of 19 collaborative researchprojects - 10 of which are new projects during FY2010.Thai Fermentation Industry Co., Ltd.Corynebacterium glutamicum strain improvementfor effective L-glutamate productionThai Better Foods Co., Ltd.Development of heat-humiditymethod for modified tapioca starchproduction at semi-industrial scaleMitr Phol Research and DevelopmentCenter Co., Ltd.Sugarcane strain improvement for salinitystress-tolerancePokphand Aquatech Co., Ltd.Development of loop mediatedisothermal amplification combinedwith the lateral flow dipstick(LAMP-LFD) technique to detectyellow head virus in shrimp andcarriersBioMedCore Inc. (Japan)Evaluating mannose-coated liposome asshrimp immune boosterSun Feed Co., Ltd.Optimization of fungal beta-mannanaseproduction and feasibility study of theenzyme for corn-based diet digestionThanapaisalApplication of enzyme-mix in a single-step desizingof and lipid stain removal from cotton fabricAsian Aqua & Animal (Thailand) Co., Ltd.Bacillus sp. strain selection and screening for digestionof organic matter and anti-microbial properties to beused in shrimp aquaculture

National Center for Genetic Engineering and Biotechnology (BIOTEC) 43CONTRACT RESEARCH AND CONSULTINGSERVICESIn 2010 BIOTEC performed 28 contract researchprojects and 14 technical consulting projects. Of these42 projects, 15 are new initiatives. Clustered by industryapplications, 6 of the new projects were under theagriculture and food sector, 5 were under the energyand environment sector, and the last 3 were underthe health and medicine sector.

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National Center for Genetic Engineering and Biotechnology (BIOTEC) 45HUMAN RESOURCEDEVELOPMENTBIOTEC places high priority on human resourcedevelopment in terms of increasing the quantity andquality of human resources in this sector as wellas upgrading and educating the workforce. Variousactivities are designed to capture different segmentsof the workforce as well as to address a varietyof objectives, ranging from providing scholarships/fellowships to organizing training workshops foracademics and industries, to organizing youthprograms.

46Annual Report 2010TECHNICAL WORKSHOPS ANDACADEMIC SEMINARSTraining Workshop on Application of Double Haploid Technique for Cucumber ImprovementThai - Franco Seminar on SmallRNA in Oil Palm ReproductiveDevelopmentTraining Workshop on BITEME:BIoTEc Microarray EnterpriseTraining Workshop on Biosafety Guidelines for WorkRelated to Modern Biotechnology or Genetic EngineeringWorkshop onBiotechnologyBusiness andRegulationTraining Workshop onBiotechnology for CattleProductionWorkshop on Gene Discoveryof Uncultured Microbes UsingMetagenomic ApproachTraining Workshop on Biotechnological Methods for Gynogenesis ofCucumber and Androgenesis of PepperWorkshop on TissueCulture of TeakWorkshops on FoodSafety Risk AssessmentWorkshop on BiosafetyPolicy and FrameworkWorkshop on ProteomicsAnalysis and Application: CaseStudies in Shrimp and SpirulinaResearch ProjectsWorkshop on OvulationInduction in CattleWorkshop on Screening Protocol for RootKnot Nematode, Meloidogyne sppWorkshop on Diagnostic Technology for Food-borne Pathogens and ChemicalContamination in Food ProductsWorkshop on Food SafetyRisk AssessmentWorkshop on Genetic Factors in BroodstockManagement for Selective Breeding Program inMarine ShrimpDuring the year, BIOTEC organized 22 trainingworkshops and seminars, as well as the 7 th NationalSymposium on Marine Shrimp, altogether attendedby 853 participants (1,858 man-days). Topics ofworkshops and seminars on offer covered variousthemes of biotechnology, e.g. plant biotechnology,animal biotechnology, biosafety and food safety andcutting-edge technologies.Public seminars with local and overseas guestspeakers are regularly organized with audiences fromuniversities, companies and government agencies.Over 40 seminars were held in 2010.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 47TONKLA ARCHEEP PROGRAM – GOVERNMENTECONOMIC STIMULUS PACKAGEIn March 2009, the Royal Thai Government launchedthe Tonkla Archeep Program, as part of thegovernment economic stimulus package to alleviatethe nation’s economic setback. Tonkla ArcheepProgram aimed to provide vocational training for theworkforce affected by economic downturn, allowingthem to re-enter the job market or start operatingtheir own small farming business with improved andnew skills.challenging and large-scale task, an alliance wasformed with the Community Enterprise Institute and itsnetwork of over 1,000 training centers across thecountry to offer on-site training to the workforce in53 provinces. The training offered was in the areasof small and micro community enterprise and holisticapproach to sufficiency economy. Over 50,000individuals were trained through this specific projectoperated by BIOTEC.BIOTEC was one of many organizations selected toprovide service to the Government in managing theproject run from June 2009 – June 2010, with totalfunding of 517 million Baht. To undertake this

48Annual Report 2010CAPACITY BUILDING ON ENERGY AND RESOURCEEFFICIENCY FOR THAI NATIVE STARCH INDUSTRYThailand is considered a major producer and exporterof tapioca and starch. Although the native starchproduction business became very competitive in therecent past, most of the native starch factories need tomake improvements on effective production processes,pollution control mechanisms, waste recycling/re-use,and energy efficiency options.Equipped with in-depth knowledge in biogas technologyand tapioca starch industrial production processes andwaste management, the Excellent Center of WasteUtilization and Management (ECoWaste), a joint unitbetween BIOTEC and King Mongkut’s University ofTechnology Thonburi, launched the project “CapacityBuilding on Energy and Resource Efficiency for Thai

National Center for Genetic Engineering and Biotechnology (BIOTEC) 49Native Starch Industry” in May 2009 with financialsupport from NSTDA and GTZ. The 2-year projectaims to improve the competitiveness of Thai nativetapioca starch industry through a practical trainingprogram targeting plant owners, managers and unitoperators. To enhance the learning process, twoguidelines for eco-efficiency improvement will bedeveloped, namely (i) best practice guide foreco-efficiency improvement in native starch industry,and (ii) management information system (MIS)guidelines for eco-efficiency for native starch industry.In 2010, the Project launched a website ThailandTapioca Starch (http://www.thailandtapiocastarch.net)to serve as a source of information and knowledgefor researchers and practitioners in the tapiocaindustry. The website is available in both Thaiand English languages. In addition to the website,newsletters have been regularly produced anddistributed amongst members of the network, whichcurrently includes more than 50 individuals, 25factories, and 20 engineering graduates. During June– August 2010, three workshops were conductedtargeting three major stakeholders, namely factorymanagers and operators; public and private entities(including environmental consulting firms); andgraduate students who plan to develop a career inthis field.

50Annual Report 2010YOUTH PROGRAMScience in Rural Schools Program (SiRS) TheProgram is dedicated to youth in the rural areas coveringthe following provinces; Maehongson, Nan, ChiangMai, Phrae, Chiang Rai, Sakon Nakhon, Pang-Ngaand Narathiwat. SiRS aims to improve the quality oflife and enhance learning capability in science for ruralstudents through activities such as science camps,workshops and science project contests. The followingactivities were organized in FY 2010:• A workshop for school administrators andteachers in Chiang Mai to train them intechniques for teaching science was held in

National Center for Genetic Engineering and Biotechnology (BIOTEC) 51November 2009.• A science camp with a banana theme washeld for students and villagers in SakonNakhon in May 2010.• A training workshop to train science teachersin Nan to create teaching-aid media for sciencecurriculum was held twice in August 2010.• A project to develop student leaders wasorganized. Two groups of leaders were chosen,one on leaders in science and the other onleaders in environmental conservation.Projects Supported by the Thai Health PromotionFoundation BIOTEC manages two projects fundedby the Thai Health Promotion Foundation, namely:• Science for Good Health The project aimsto supplement teachers in the target areawith heath-related scientific knowledge andengage students in science activities. PhaseI of the Project, implemented between July2007- March 2009, covered 224 schools and666 teachers. As a result of the project, 130science camps were organized and 561 scienceprojects were initiated. With the success ofPhase I, the Thai Health Promotion Foundationextended the project into Phase II until2011. Under Phase II, seven workshops wereconducted, with participation by science teachersfrom over 100 schools. The activities generated269 science projects, and a science camp washeld in Chiang Rai Province, with participationof 14 schools entering the science competition.• Development of Local Science Curriculumfor Southern Border Provinces A set of elevenchildren’s books were developed for students inthis region under the theme “Living a Happyand Healthy Life”. The content was developedby BIOTEC together with local school teachersand students dealing with health issues,incorporating the wisdom and knowledgein living a healthy life. The books weresubsequently reviewed and endorsed byacademic experts. 5,000 copies of the bookswere printed and introduced in the secondsemester of 2010 academic year.

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National Center for Genetic Engineering and Biotechnology (BIOTEC) 53PUBLIC AWARENESSStriving to make Thailand a knowledge-basedeconomy, one of BIOTEC’s missions is to createpublic awareness in biotechnology, especially relatingbiotechnology and life sciences to everyday living.The mission is carried out by channeling informationthrough various available media such as internet andtelevision, as well as children’s books.

54Annual Report 2010BOOKSWonderful World A children’s book on geneticallymodifiedmicroorganisms was released in 2010. Thisis the fourth book in the series “Wonderful World” toeducate children on basic biology and biotechnology,with simple content, illustrations and daily-life examples.Three previous books were on microorganisms, DNAand biodiversity.Living a Happy and Healthy Life Living a Happyand Healthy Life is a set of eleven children booksprepared under the project “Development of LocalScience Curriculum for Southern Border Provinces”funded by the Thai Health Promotion Foundation.The content was developed by BIOTEC together withlocal school teachers and students dealing with healthissues and incorporating the wisdom and knowledgein living a healthy life. 5,000 copies of the books wereprinted and introduced to schools in the four southernborder provinces during the 2010 academic year.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 55WEBSITETELEVISION PROGRAMA new version of the Thai-language corporate websitewas launched in January 2010. In addition to providinginformation on the organization itself, the BIOTECwebsite is filled with knowledge and informationon biotechnology and its application, catering to awide-spectrum of audiences from children to adults,from layman to technical persons to entrepreneurs.For educational purposes, an update on world-widebiotechnology in the form of articles is regularlyposted on the website.NSTDA produces documentaries for TV broadcasting,some of which cover BIOTEC activities:Short Documentary The 3-minute documentary isshown as part of the Science TV Program, aired onChannel 9. Documentaries introducing biotechnologytopics include: BIOTEC researcher - Winner of L’OrealFor Women in Science, microtube gel test kit andBIOTEC researcher - Winner of Young ScientistAward.Chowwit Chit Chowbaan TV Program Chowwit ChitChowbaan is a 1-hr documentary aiming to introduceresearch that can have an impact on the public,especially farmers. The show is aired every Fridayfrom 11:05-11:55 on the Thai PBS Channel andon public holidays from 11:00-11:30 on Channel 9.Seventeen episodes on biotechnology topics wereproduced and aired in FY 2010, on the following topics:rice seed production technology, sugarcane varietyimprovement through biotechnology, germinated brownrice, fish sausage, biogas technology, screening fordisease-resistant cucumber, surimi: fish-based foodproduct and feed additive for growing marineanimals.

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National Center for Genetic Engineering and Biotechnology (BIOTEC) 57INTERNATIONALCOLLABORATIONThe BIOTEC International Cooperation Program aimsto capitalize on international linkages to make BIOTECand Thailand a regional leader in the field ofbiotechnology. In so doing, the Center has developedclose linkages with overseas organizations at thebilateral, multilateral and regional levels. The goal isachieved through activities such as establishing formalcollaborative agreements, organizing joint scientificseminars with international partners, establishingand organizing an annual meeting of the BIOTECInternational Advisory Board and showcasing BIOTECand Thailand’s biotechnology in international events.

58Annual Report 2010FOSTERING RESEARCH AND ACADEMICCOLLABORATIONBIOTEC entered into MOUs with 3 institutes to promoteresearch collaboration and staff / student exchange:• Temasek Polytechnic, Singapore – BIOTEC startedhosting students from Temasek Polytechnic inBIOTEC laboratories in 2007. The program grewsteadily over the years, leading to formalization ofthe collaboration through an MOU in 2010.• Queen’s University of Belfast, Northern Ireland– The first joint research project started in2010 under this MOU framework involves thedevelopment of multiplex detection for plantdiseases in melon using antibody array technologyin a multiwell-plate format and multiplex SPR.• University of North Texas Health Science Centerat Fort Worth, USA – Under this MOU framework,BIOTEC has licensed the use of iCollect, software

National Center for Genetic Engineering and Biotechnology (BIOTEC) 59developed by the BIOTEC Information SystemsLaboratory to support the laboratory’s inventorymanagement of biological collections for academicpurposes at UNT.To continue collaboration with existing partners, BIOTECrenewed MOUs with the following organizations in2010:• Korea Research Institute for Bioscience andBiotechnology (KRIBB) -- The collaboration withKRIBB started in 2004, with a regular jointseminar to enhance research collaboration.Enzyme technology is one of the active researchtopics between the two parties.• Atma Jaya Catholic University, Indonesia - Thefirst MOU with Atma Jaya Catholic University wasformalized in 2005. Since then, BIOTEC labsregularly host students for research internship.• Institute of Biotechnology (IBT), Vietnam - TheMOU signed in 2005 led to a joint research projecton “A/H5N1 influenza virus in Vietnam andThailand: Molecular Epidemiology, Diagnosis andVaccines” funded by BIOTEC and the VietnameseMinistry of Science and Technology since March2008. A new project on “Large-scale sequencingand bioinformatics analysis of express sequencetags (ESTs) and partial genomic sequencesof the giant tiger shrimp (Penaeus monodon)using 454 pyrosequencing technology” has beeninitiated through two workshops held in Marchand September 2010 in Thailand and Vietnam,respectively. The new project is scheduled tocommence in 2011, with funding from BIOTECand the Vietnamese Ministry of Science andTechnology.Cultural Organization (UNESCO) Bangkok Office.• UK-Brazil-Thailand Joint Seminar on AgriculturalBiotechnology” on 22 - 24 February 2010, organizedin collaboration with the UK Biotechnology andBiological Sciences Research Council (BBSRC) andthe British High Commission in Singapore.Two joint seminars were organized:• The 5th BIOTEC and KRIBB Joint Symposiumon Bioresources Utilization and Management on16 October 2009, organized in collaboration withthe Korea Research Institute for Bioscience andBiotechnology (KRIBB)• Symposium on “Malaria R&D Financing, PolicyInnovation and Emerging Drug Resistance inSoutheast Asia - Perspectives from the Local andGlobal” on 9 - 10 November 2009, organized incollaboration with the University of Cambridge, theUK Economic and Social Research Council (ESRC),United Nations Educational and Scientific and

60Annual Report 2010STRENGTHENING RICE BREEDING PROGRAM INMEKONG REGIONThe Molecular Rice Breeding Program for the MekongRegion was launched in 2004 by the Rice GeneDiscovery Unit aiming to promote the implementationof marker aided selection (MAS) into the current ricebreeding programs in the Mekong region, particularlyThailand, Laos, Cambodia and Myanmar through acomprehensive hands-on training program and sharingof genomic information (genetic data for several traitsand the molecular markers for several genes) andresearch facilities. The program was designed to addressthe limitation of short-term workshops which cannotleave participants with full understanding of thetechnology and laboratory skills, as well as the limitationof research facilities in these countries. To ensurethe benefit of this program to each Mekong country,participants perform the MAS breeding to improvetheir own rice varieties.The program obtained research funding from theGeneration Challenge Program (GCP) in 2007-2008under the project title “Community of Practices -Concept Applied to Rice Production in the MekongRegion: Quick conversion of popular rice varietieswith emphasis on drought, salinity and grain qualityimprovement”In November 2009, the continuation of this projecttitled “Community of Practices – Strengthening ricebreeding program using genotyping building strategyand improving phenotyping capacity for biotic andabiotic stresses in the Mekong region” was approvedfor funding from GCP.Partners in this Program include the CambodianAgricultural Research and Development Institute,(CARDI), National Agricultural and Forestry Institute(NAFRI) from Laos, the Department of AgriculturalResearch (DAR) from Myanmar and the UbonRatchathani University from Thailand.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 61REGIONAL R&D AND TRAINING HUBWith over two decades of operation, BIOTEC has builtconfidence in its high-caliber researchers and R&Dinfrastructure and attempts to utilize this expertise andfacilities to build up research and technical capacityfor the world community. The Center provides foreignstudents and researchers in the field of biotechnologyand life sciences with opportunities to have hands-onexperience conducting research in a professionalenvironment.• Human Resource Development Program forNeighboring Countries Initiated in 2001, theProgram provides fellowships to young scientistsfrom developing countries in the Asia-Pacificregion to work in BIOTEC laboratories for 3-6months. In FY 2010, out of 89 applicants from10 countries, 12 fellowships were provided toresearchers from Myanmar, Lao PDR, Vietnam,the Philippines, Mongolia and Indonesia. Thesurvey conducted on past alumni, reaching 100alumni this year, has demonstrated the successof the program. At least twenty eight alumniobtained scholarships to further their educationoutside their home countries such as Australia,Austria, Japan, Korea, the Netherlands, thePhilippines and USA. Most of them are makingprogress in their research work and continuefocusing on their research career paths.• International Exchange Program This Programis designed for hosting foreign students to havehands-on experience conducting research in aprofessional environment, as well as for foreignresearchers who would like to obtain some trainingin particular areas. Fifty foreign students andresearchers, both new and on-going, were hostedin FY2010 from institutes such as Atma JayaCatholic University, Indonesia; University of Kent,UK; Temasek Polytechnic, Singapore; NanyangPolytechnic, Singapore; University of Monpellier,France. This year marked the first participation bythe University of Liverpool, sending 5 students tojoin the Program.

62Annual Report 2010THAILAND PAVILION AT BIO INTERNATIONALCONVENTION 2010BIOTEC, in collaboration with the Thai public andprivate sectors, organized the Thailand Pavilion at the2010 Bio International Convention held in Chicago on3 – 6 May 2010.This year, the Thailand Pavilion focused on Green &Clean Technology to showcase the nation’s abundantpotential in utilizing its natural resources and homegrownstrong R&D base to develop green technologyto support the nation’s growing economy.Highlights of the Thailand Pavilion also included thelaunch of a publication on Life Sciences in Thailand,as well as a mini–forum with talks covering varioustopics such as “Biotechnology R&D in Thailand” and“Biotech Business Opportunities in Asia”, with speakersfrom the Board of Investment, Thailand Science Parkand some foreign companies in Thailand.Thailand was among fifteen countries selected tomake a presentation in the International Case Studiestrack during the Convention. The presentation showedworking mechanisms to synergize the efforts andidentify opportunities in R&D in Thailand, using anR&D joint venture between Thailand’s BIOTEC andNanoDetection Technology, an American company, asan example.The Thai delegation consisted of 13 organizations,including government authorities in R&D andinvestment, academic institutes as well as local andmultinational companies.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 63LIFE SCIENCES IN THAILANDLife Sciences in ThailandBIOTEC spearheaded the publication of Life Sciencesin Thailand, the first ever supplement with a majorinternational magazine devoted to biotechnology andlife sciences in Thailand, providing a comprehensivelook into life sciences policy, industry and researchin Thailand. 80,000 copies were printed for globaldistribution. The supplement was unveiled at BioInternational Convention 2010 held during 3-6 May2010 in Chicago, USA, and released with the Mayissue of the Scientist.Joining force with BIOTEC to finance this publicationwere various scientific organizations, universities,government agencies and the private sector, bothlocal and multinational.The publication is available for download from thewebsites of BIOTEC, the Scientist and sponsors.

64Annual Report 2010

National Center for Genetic Engineering and Biotechnology (BIOTEC) 65IMPACT OF BIOTEC’SOUTPUTBIOTEC constantly monitors and assesses the impactof its output towards social and economic development.Every year, BIOTEC selects a number of technologytransfer projects for detailed impact study. The impactis measured in the form of income generated bythe clients out of such products and technologies,and also quantified from parameters such as importsubstitution and employment generation.

66Annual Report 2010EXAMPLES OF IMPACT FROM PROJECTS INAGRICULTURE AND FOODMicropropagation of Disease-free Sugarcane During2003-2005, in response to a request from MitrPhol Research and Development Center Co., Ltd., aresearch and development arm of the Mitr Phol SugarGroup, BIOTEC provided technical assistance to thecompany in establishing a tissue culture laboratory forsugarcane. Following this initial collaboration, BIOTECprovided continuous assistance to the company to helpbuild up their research capability through training andresearch collaboration and to help the company expandits sugarcane production. During the productionyear 2009/2010, the company was able to earn anadditional 32 million Baht as a result of the productionexpansion. The expansion also created an additional3.7 million Baht in employment value.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 67Germplasm Evaluation of Sugarcane Elite Linesacross the Country In 2006, BIOTEC-NSTDA launcheda project to evaluate sugarcane elite lines developedby 5 major sugarcane research centers through fieldtrials in various areas by working in partnership withresearch stations across the country. The result offield trials assured the quality of the lines amongfarmers, resulting in a plantation expansion of newlines in 300,000 rai (48,000 hectares) equivalent to 3billion Baht in income earned from the additional yieldthese elite lines can provide.Virus Resistant Hybrid Okra, UNIH109 In 2004,BIOTEC collaborated with Uniseeds Co., Ltd. todevelop virus resistant okra. Through the collaboration,the company was able to develop a new variety,UNIH109, yielding 2,500 – 3,000 kg/rai (15.63 – 18.75tonnes/hectare), while the average yield was 1,000kg/rai (6.25 tonnes/hectare). In 2010, the companyearned 3.5 million Baht in sales value for UNIH109,and the value of okra production, estimated from theseed volume sold, would reach 138.3 billion Baht forthe farmers.Air Filter for Tissue Culture Laboratory BIOTECresearchers have developed an air filter forgreenhouses or tissue culture containers, creatingan environmental condition to allow for air andmoisture exchange in the tissue culture process.The technology was licensed to Nisapan Co., Ltd., atissue culture business, in March 2008 for 5 years. Thecompany made 0.3 million Baht selling this productin 2010. The air filter helps reduce plantlet loss duringthe acclimatization period by 30%, the survivingplantlets being equivalent to 3 million Baht. As theequivalent imported product costs 5-6 times more,this locally-made air filter was able to save thecountry 1.96 million Baht in foreign exchange costs.Rapid Test Kit for Sugarcane White Leaf DiseaseBIOTEC, NSTDA and Mitr Phol Research andDevelopment Center Co., Ltd., have jointly developeda rapid test kit for phytoplasma infection, which isthe cause of white leaf disease in sugarcane. Thetest kit allows farmers to screen for phytoplasma-freecane stalks before planting, which will not only reducelosses, but will also minimize the spread of disease tohealthy plants. The test kit has been sold in Thailand,as well as neighboring countries such as Laos andCambodia, with a sales value of 1.15 million Baht.Steroid-based Synchronization of Ovulation Protocoland Fixed Time Artificial Insemination BIOTECresearchers have transferred the technology to farmersfor ovulation synchronization, using hormone andartificial insemination, through training workshopsand follow-up consultancy. The training and technicalassistance has enabled the farmers to induce thepregnancy in 3,164 cows which previously havereproductive system problems, adding 20,000 Bahtto the value of each cow. As cows reach lactation,additional income from milk production can also beearned. The total monetary impact is estimated to beover 270 million Baht.

68Annual Report 2010EXAMPLES OF IMPACT FROM PROJECTS INHEALTH AND MEDICINECD4+ Lymphocytes Enumeration Assay The assayfor enumeration of CD4+ lymphocytes was developedby the Biomedical Technology Research Center, a jointlaboratory between BIOTEC and Chiang Mai University.The assay does not require an expensive flowcytometer, but instead uses a hematoanalyzer, whichis common equipment in general hospitals. Thetechnology was licensed to i+MED Laboratories Co.,Ltd. In 2010, i+MED Laboratories earned 0.24 millionBaht in revenue. This locally-made product was ableto save the country 0.53 million Baht from importsand the cost of eliminating the purchase of flowcytometers is estimated at 0.26 million Baht.Test for Red Cell Antigen-Antibody DetectionBIOTEC-NSTDA provided a research grant to KhonKaen University to develop a test for red cellantigen-antibody detection. The test can be massproduced domestically at a relatively low cost, allowingfor a cost of 5-10 Baht/test, while the imported testcan cost up to 25 Baht. The technology was licensedto Innov (Thailand) Co., Ltd. and the product wasreleased in the market in January 2010, contributing0.94 million Baht in sales to the company. The savingfrom import costs for this product is estimated at 3.7million Baht.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 69EXAMPLES OF IMPACT FROM PROJECTS INBIORESOURCES UTILIZATION AND ENVIRONMENTPromotion of Biogas Technology in CassavaStarch Industry Biogas technology for wastewatertreatment and methane production was developed byEcoWaste and implemented in 3 factories, resulting in43.62 million Baht in energy savings and unaccountedenvironmental benefits.Pentosanase Production Technology BIOTEC hasdeveloped the production technology which usesthe pentosanase-producing microorganism carefullyscreened from the BIOTEC Culture Collection and theprocess to achieve a final product in powder form.The technology was licensed to Asia Star AnimalHealth Co., Ltd. (ASAH) to manufacture and distributepentosanase as a feed additive. The company earned2.4 million Baht in sales from the product, savingthe country 4.8 million Baht/year from importing thisenzyme. The impact from healthier animals, bettermeat quality and the saving on feed is estimated tobe 22.35 million Baht.Promotion of Biogas Technology in Agri-food IndustryThe fixed-film type of biogas technology, designed byEcoWaste, was installed in 2 factories making dry fruitproducts. The value of waste treatment cost saving,energy saving and electricity generation totaled 30million Baht.Biogas Technology in Palm Oil Industry The researchand engineering team at EcoWaste has designed andinstalled a combination of fixed-bed and sludge-bedreactor, suitable for wastewater from palm oil productionin Thachana Palm Oil Co., Ltd, adding to an existingreactor. The new reactor helps increase the stabilityof the wastewater systems of the factory, as well asaccommodating a higher loading rate. Biogas producedis valued at 1.2 million Baht/year.Development of Zero Waste Discharge CassavaStarch Factory Researchers and engineers fromEcoWaste have been continuously developingtechnologies to improve production processes, pollutioncontrol mechanisms, waste recycling/re-use andenergy efficiency options. In 2010, researchers wereable to make an improvement on an extractor andrecover more starch from water discharged froma separator at Cholcharoen Factory, resulting in anadditional 33.34 million Baht from the resultant yieldincrease and savings.

APPENDICESList of PublicationsList of Patents and Petty PatentsHonors and AwardsExecutives and Management Team

72Annual Report 2010LIST OF PUBLICATIONS1. Abdel-Wahab, M.A., Pang, K.-L., Nagahama, T., Abdel-Aziz,F.A. and Jones, E.B.G. (2010). Phylogenetic evaluationof anamorphic species of Cirrenalia and Cumulosporawith the description of eight new genera and four newspecies. Mycological Progress, doi: 10.1007/s11557-010-0661-x.2. Abdulla, M.A., Ahmed, I., Assawamakin, A., Bhak, J.,Brahmachari, S.K., Calacal, G.C., Chaurasia, A., Chen,C.H., Chen, J., Chen, Y.T., Chu, J., Cutiongco-de la Paz,E.M.C., De Ungria, M.C.A., Delfin, F.C., Edo, J., Fuchareon,S., Ghang, H., Gojobori, T., Han, J., Ho, S.F., Hoh, B.P.,Huang, W., Inoko, H., Jha, P., Jinam, T.A., Jin, L., Jung,J., Kangwanpong, D., Kampuansai, J., Kennedy, G.C.,Khurana, P., Kim, H.L., Kim, K., Kim, S., Kim, W.Y.,Kimm, K., Kimura, R., Koike, T., Kulawonganunchai, S.,Kumar, V., Lai, P.S., Lee, J.Y., Lee, S., Liu, E.T., Majumder,P.P., Mandapati, K.K., Marzuki, S., Mitchell, W., Mukerji,M., Naritomi, K., Ngamphiw, C., Niikawa, N., Nishida, N.,Oh, B., Oh, S., Ohashi, J., Oka, A., Ong, R., Padilla,C.D., Palittapongarnpim, P., Perdigon, H.B., Phipps, M.E.,Png, E., Sakaki, Y., Salvador, J.M., Sandraling, Y., Scaria,V., Seielstad, M., Sidek, M.R., Sinha, A., Srikummool, M.,Sudoyo, H., Sugano, P., Suryadi, H., Suzuki, Y., Tabbada,K.A., Tan, A., Tokunaga, K., Tongsima, S., Villamor, L.P.,Wang, E., Wang, Y., Wang, H., Wu, J.Y., Xiao, H., Xu,S., Yang, J.O., Shugart, Y.Y., Yoo, H.S., Yuan, W., Zhao,G., Zilfalil, B.A. and Indian Genome Variation Consortium.(2009). Mapping Human Genetic Diversity in Asia. Science,326(5959), 1541-1545.3. Aewsiri, T., Benjakul, S., Visessanguan, W., Wierenga,P.A. and Gruppen, H. (2010). Antioxidative activity andemulsifying properties of cuttlefish skin gelatin-tannic acidcomplex as influenced by types of interaction. InnovativeFood Science and Emerging Technologies, doi:10.1016/j.ifset.2010.04.001.4. Am-In, S., Limtong, S., Yongmanitchai, W. and Jindamorakot,S. (2010). Candida andamanensis sp. nov., Candidalaemsonensis sp. nov., and Candida ranongensis sp. nov.,three anamorphic yeast species isolated from estuarine watersin a mangrove forest in Ranong Province, Thailand.International Journal of Systematic and EvolutionaryMicrobiology, doi:10.1099/ijs.0.022038-0.5. Amparyup, P., Wiriyaukaradecha, K., Charoensapsri, W.and Tassanakajon, A. (2010). A clip domain serineproteinase plays a role in antibacterial defense but isnot required for prophenoloxidase activation in shrimp.Developmental and Comparative Immunology, 34(2),168-176.6. Angthong, P., Watthanasurorot, A., Klinbunga, S., Ruangdej,U., Söderhäll, I. and Jiravanichpaisal, P. (2010). Cloningand characterization of a melanization inhibition protein(PmMIP) of the black tiger shrimp, Penaeus monodon.Fish and Shellfish Immunology, 29(3), 464-468.7. Anuchapreeda, S., Tima, S., Duangrat, C. and Limtrakul,P. (2008). Effect of pure curcumin, demethoxycurcumin,and bisdemethoxycurcumin on WT1 gene expressionin leukemic cell lines. Cancer Chemotherapy andPharmacology, 62(4), 585-594.8. Arunpanichlert, J., Rukachaisirikul, V., Sukpondma, Y.,Phongpaichit, S., Tewtrakul, S.,Rungjindamai, N. andSakayaroj, J. (2010). Azaphilone and isocoumarinderivatives from the endophytic fungus Penicilliumsclerotiorum PSU-A13. Chemical and PharmaceuticalBulletin, 58(8), 1033-1036.9. Banyai, W., Kirdmanee, C., Mii, M. and Supaibulwatana, K.(2010). Overexpression of farnesyl pyrophosphate synthase(FPS) gene affected artemisinin content and growth ofArtemisia annua L. Plant Cell Tissue and Organ Culture,doi:10.1007/s11240-010-9775-8.10. Benjakul, S., Thiansilakul, Y., Visessanguan, W., Roytrakul,S., Kishimura, H., Prodpran, T. and Meesane, J. (2010).Extraction and characterisation of pepsin-solubilised collagensfrom the skin of bigeye snapper (Priacanthus tayenus andPriacanthus macracanthus). Journal of the Science ofFood and Agriculture, 90(1), 132-138.11. Benjakul, S., Yarnpakdee, S., Visessanguan, W. andPhatcharat, S. (2010). Combination effects of wheyprotein concentrate and calcium chloride on the propertiesof goatfish surimi gel. Journal of Texture Studies, 41(3),341-357.12. Bertoft, E., Laohaphatanalert, K., Piyachomkwan, K. andSriroth, K. (2010). The fine structure of cassava starchamylopectin. Part 2: Building block structure of clusters.International Journal of Biological Macromolecules, 47(3),325-335.13. Boondaeng, A., Suriyachadkun, C., Ishida, Y., Tamura, T.,Tokuyama, S. and Kitpreechavanich, V. (2010). Herbidosporasakaeratensis sp. nov., isolated from Thailand and reclassificationof Streptosporangium claviforme as alater synonym of Herbidospora cretacea. InternationalJournal of Systematic and Evolutionary Microbiology,doi:10.1099/ijs.0.024315-0.14. Boonmak, C., Limtong, S., Jindamorakot, S., Am-In, S.,Yongmanitchai, W., Suzuki, K.I., Nakase, T. and Kawasaki,H. (2010). Candida xylanilytica sp. nov., a xylan degradingyeast species isolated from Thailand. International Journalof Systematic and Evolutionary Microbiology, doi:10.1099/ijs.0.021873-0.15. Boonyos, P., Soonsanga, S., Boonserm, P. and Promdonkoy,B. (2010). Role of cysteine at positions 67, 161 and 241of a Bacillus sphaericus binary toxin BinB. Journal ofBiochemistry and Molecular Biology, 43(1), 23-28.16. Buaban, B., Inoue, H., Yano, S., Tanapongpipat, S., Ruanglek,V., Champreda, V., Pichyangkura, R., Rengpipat, S. andEurwilaichitr, L. (2010). Bioethanol production from ball milledbagasse using an on-site produced fungal enzyme cocktailand xylose-fermenting Pichia stipitis. Journal of Bioscienceand Bioengineering, doi: 10.1016/j.jbiosc.2009.12.003.17. Bunterngsook, B., Kanokratana, P., Thongaram, T.,Tanapongpipat, S., Uengwetwanit, T., Rachdawong, S.,

National Center for Genetic Engineering and Biotechnology (BIOTEC) 73Vichitsoonthonkul, T. and Eurwilaichitr, L. (2010).Identification and Characterization of Lipolytic Enzymesfrom a Peat-Swamp Forest Soil Metagenome. BioscienceBiotechnology and Biochemistry, 74(9), 1848-1854.18. Bunyapaiboonsri, T., Yoiprommarat, S., Srikitikulchai, P.,Srichomthong, K. and Lumyong, S. (2010). Oblongolidesfrom the Endophytic Fungus Phomopsis sp. BCC 9789.Journal of Natural Products, 73(1), 55-59.19. Cerenius, L., Babu, R., Söderhäll, K. and Jiravanichpaisal,P. (2010). In vitro effects on bacterial growth ofphenoloxidase reaction products. Journal of InvertebratePathology, 103(1), 21-23.20. Chaiklahan, R., Chirasuwan, N., Siangdung, W.,Paithoonrangsarid, K. and Bunnag, B. (2010). Cultivationof Spirulina platensis using pig wastewater in asemi-continuous process. Journal of Microbiology andBiotechnology, 20(3), 609-614.21. Chaiprasert, A., Krajaejun, T., Pannanusorn, S.,Prariyachatigul, C., Wanachiwanawin, W., Sath patayavongs,B., Juthayothin, T., Smittipat, N., Vanittanakom, N. andChindamporn, A. (2010). Pythium insidiosum Thai isolates:molecular phylogenetic analysis. Asian Biomedicine, 3(6),623-633.22. Chaivisuthangkura, P., Longyant, S., Hajimasalaeh, W.,Sridulyakul, P., Rukpratanporn, S. and Sithigorngul, P.(2010). Improved sensitivity of Taura syndrome virusimmunodetection with a monoclonal antibody against therecombinant VP2 capsid protein. Journal of Virological Methods,163(2), 433-439.23. Chaivisuthangkura, P., Longyant, S., Rukpratanporn, S.,Srisuk, C., Sridulyakul, P. and Sithigorngul, P. (2010).Enhanced white spot syndrome virus (WSSV) detectionsensitivity using monoclonal antibody specific toheterologously expressed VP19 envelope protein.Aquaculture, 299(1-4), 15-20.24. Chaivisuthangkura, P., Srisuk, C., Rukpratanporn, S.,Longyant, S., Sridulyakul, P. and Sithigorngul, P. (2010).Rapid and sensitive detection of Penaeus monodonnucleopolyhedrovirus by loop-mediated isothermalamplification. Journal of Virological Methods, 162(1-2),188-193.25. Chamroensaksri, N., Tanasupawat, S., Akaracharanya,A., Visessanguan, W., Kudo, T. and Itoh, T. (2009).Salinivibrio siamensis sp. nov., from fermented fish(pla-ra) in Thailand. International Journal of Systematicand Evolutionary Microbiology, 59(4), 880-885.26. Chamroensaksri, N., Tanasupawat, S., Akaracharanya, A.,Visessanguan, W., Kudo, T. and Itoh, T. (2010). Gracilibacillusthailandensis sp. nov., from fermented fish (pla-ra).International Journal of Systematic and EvolutionaryMicrobiology, 60(4), 944-948.27. Chanama, M., Chanama, S., Shaw, P.J., Chitnumsub, P.,Leartsakulpanich, U. and Yuthavong, Y. (2010). Formationof catalytically active cross-species heterodimers ofthymidylate synthase from Plasmodium falciparum andPlasmodium vivax. Molecular Biology Reports, doi:10.1007/s11033-010-0199-7.28. Chareonlimkun, A., Champreda, V., Shotipruk, A.and Laosiripojana, N. (2010). Catalytic conversion ofsugarcane bagasse, rice husk and corncob in thepresence of TiO 2, ZrO 2and mixed-oxide TiO 2-ZrO 2underhot compressed water (HCW) condition. BioresourceTechnology, 101(11), 4179-4186.29. Chareonlimkun, A., Champreda, V., Shotipruk, A. andLaosiripojana, N. (2010). Reactions of C 5and C 6-sugars,cellulose, and lignocellulose under hot compressed water(HCW) in the presence of heterogeneous acid catalysts.Fuel, 89(10), 2873-2880.30. Charerntantanakul, W. and Kasinrerk, W. (2010).Interleukin-10 antisense oligodeoxynucleotide suppressesIL-10 expression and effects on proinflammatorycytokine responses to porcine reproductive andrespiratory syndrome virus. Viral Immunology, 23(4),425-435.31. Charlermroj, R., Gajanandana, O., Barnett, C., Kirtikara,K. and Karoonuthaisiri, N. (2010). A chemiluminescentantibody array system for detection of foodbornepathogens in milk. Analytical Letters, doi:10.1080/00032719.2010.511736.32. Charoensapsri, W., Amparyup, P., Hirono, I., Aoki, T. andTassanakajon, A. (2010). PmPPAE2, a new class ofcrustacean prophenoloxidase (proPO)-activating enzymeand its role in PO activation. Developmental andComparative Immunology, doi:10.1016/j.dci.2010.09.002.33. Charoenthaikij, P., Jangchud, K., Jangchud, A.,Piyachomkwan, K., Tungtrakul, P. and Prinyawiwatkul,W. (2009). Germination conditions affect physicochemicalproperties of germinated brown rice flour. Journal ofFood Science, 74(9), c658-c665.34. Cha-um S. and Kirdmanee, C. (2010). Salt tolerancescreening in six maize (Zea mays L.) genotypes usingmultivariate cluster analysis. Philippine AgriculturalScientist, 93(2), 156-164.35. Cha-um S., Ashraf, M. and Kirdmanee, C. (2010).Screening upland rice (Oryza sativa L. ssp. indica)genotypes for salt-tolerance using multivariate clusteranalysis. African Journal of Biotechnology, 9(30), 4731-4740.36. Cha-um, S. and Kirdmanee, C. (2009). Proline Accumulation,Photosynthetic Abilities and Growth Characters ofSugarcane (Saccharum officinarum L.)Plantlets inResponse to Iso-Osmotic Salt and Water-Deficit Stress.Agricultural Science in China, 8(1), 51-58.37. Cha-um, S. and Kirdmanee, C. (2010). Effect of glycinebetaineon proline, water use and photosynthetic efficienciesand growth of rice seedlings under salt stress. TurkishJournal of Agriculture and Forestry, doi:10.3906/tar-0906-34.38. Cha-um, S. and Kirdmanee, C. (2010). Effects of waterstress induced by sodium chloride and mannitol onproline accumulation, photosynthetic abilities and growthcharacters of eucalyptus (Eucalyptus camaldulensisDehnh.). New Forests, doi:10.1007/s11056-010-9204-1.39. Cha-um, S., Boriboonkaset, T., Pichakum, A. and Kirdmanee,C. (2009). Multivariate physiological indices for salttolerant classification in indica rice (Oryza sativa L. spp.

74Annual Report 2010indica). General and Applied Plant Physiology, 35(1-2),75-87.40. Cha-Um, S., Nhung, N.T.H. and Chalermpol, K. (2010).Effect of mannitol- and salt-induced iso-osmotic stress onproline accumulation, photosynthetic abilities and growthcharacters of rice cultivars (Oryza sativa L. spp. Indica).Pakistan Journal of Botany, 42(2), 927-941.41. Cha-Um, S., Siringam, K., Juntawong, N. andKirdmanee, C. (2010). Water relations, pigmentstabilization, photosynthetic abilities and growthimprovement in salt stressed rice plants treated withexogenous potassium nitrate application. InternationalJournal of Plant Production, 4(3), 187-198.42. Cha-um, S., Supaibulwatana, K. and Kirdmanee, C.(2009). Comparative Effects of Salt Stress and ExtremepH Stress Combined on Glycinebetaine Accumulation,Photosynthetic Abilities and Growth Characters of TwoRice Genotypes.Rice Science, 16(4), 274-282.43. Cha-Um, S., Takabe, T. and Kirdmanee, C. (2010). 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78Annual Report 2010130. Pengsuk, C., Longyant, S., Rukpratanporn, S., Chaivisuthangkura,P., Sridulyakul, P. and Sithigorngul, P. (2010).Development of monoclonal antibodies for simpledetection and differentiation of Vibrio mimicus from V.cholerae and Vibrio spp. by dot blotting. Aquaculture,300(1-4), 17-24.131. Phannachet, K., Raksat, P., Limvuttegrijeerat, T. andPromdonkoy, B. (2010). Production and Characterizationof N- and C-terminally Truncated Mtx2: a MosquitocidalToxin from Bacillus sphaericus. Current Microbiology, doi:10.1007/s00284-010-9651-0.132. Piccinato, C.A., Sartori, R., Sangsritavong, S., Souza, A.H.,Grummer, R.R., Luchini, D. and Wiltbank, M.C. (2010).In vitro and in vivo analysis of fatty acid effects onmetabolism of 17β-estradiol and progesterone in dairycows. Journal of Dairy Science, 93(5), 1934-1943.133. Pinruan, U., Rungjindamai, N., Choeyklin, R., Lumyong,S., Hyde, K.D. and Jones, E.B.G. (2010). 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National Center for Genetic Engineering and Biotechnology (BIOTEC) 79and Phongpaichit, S. (2010). Phylogenetic diversity ofendophyte assemblages associated with the tropicalseagrass Enhalus acoroides in Thailand. Fungal Diversity,doi10.1007/s13225-009-0013-9.152. Saksmerprome, V., Charoonnart, P., Gangnonngiw,W. and Withayachamnankul, B. (2009). A novel andinexpensive application of RNAi technology to protectshrimp from viral disease. Journal of Virological Methods,162(1-2), 213-217.153. Saksmerprome, V., Puiprom, O., Noonin, C. and Flegel,T.W. (2010). Detection of infectious hypodermal andhaematopoietic necrosis virus (IHHNV) in farmed AustralianPenaeus monodon by PCR analysis and DNA sequencing.Aquaculture, 298(3-4), 190-193.154. Sangseethong, K., Termvejsayanon, N. and Sriroth,K. (2010). Characterization of physicochemical propertiesof hypochlorite- and peroxide-oxidized cassava starches.Carbohydrate Polymers, 82(2), 446-453.155. 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Different properties of electrospun fibrousscaffolds of separated heavy-chain and light-chainfibroins of Bombyx mori. International Journal ofBiological Macromolecules, 46(5), 493-501.202. Wang, H.W., Noland, C., Siridechadilok, B., Taylor,D.W., Ma, E., Felderer, K., Doudna, J.A. and Nogales,E. (2009). Structural insights into RNA processing bythe human RISC-loading complex. Nature Structural andMolecular Biology, 16(11), 1148-1153.203. Wangchuk, P., Bremner, J.B., Samten, Rattanajak, R. andKamchonwongpaisan, S. (2009). Antiplasmodial agentsfrom the Bhutanese medicinal plant Corydalis calliantha.Phytotherapy Research, doi 10.1002/ptr.2893.204. Wangchuk, P., Bremner, J.B., Samten, Skelton, B.W.,White, A.H., Rattanajak, R. and Kamchonwongpaisan, S.(2010). Antiplasmodial activity of atisinium chloride fromthe Bhutanese medicinal plant, Aconitum orochryseum.Journal of Ethnopharmacology, 130(3), 559-562.205. Wanichananan, P., Teerakathiti, T., Roytrakul, S., Kirdmanee,C. and Peyachoknagul, S. (2010). A highly efficientmethod for Agrobacterium mediated transformation inelite rice varieties (Oryza sativa L. spp. indica). AfricanJournal of Biotechnology, 9(34), 5488-5495.206. Wanitchang, A., Kramyu, J. and Jongkaewwattana, A. (2010).Enhancement of Reverse Genetics-derived Swine-OriginH1N1 Influenza Virus Seed Vaccine Growth by Inclusionof Indigenous Polymerase PB1 Protein. Virus Research,147(1), 145-148.207. Washio, K., Lim, S.P., Roongsawang, N. and Morikawa, M.(2010). Identification and Characterization of the GenesResponsible for the Production of the Cyclic LipopeptideArthrofactin by Pseudomonas sp. MIS38. BioscienceBiotechnology and Biochemistry, 74(5), 992-999.208. Watthanasurorot, A., Jiravanichpaisal, P., Sderhll, I. andSderhll, K. (2010). A gC1qR prevents WSSV replicationin the freshwater crayfish Pacifastacus leniusculus.Journal of Virology, doi:10.1128/JVI.01045-10.209. 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Analysis of a thermophilic lignocellulose degradingmicrobial consortium and multi-species lignocellulolyticenzyme system. Enzyme and Microbial Technology, 47(6),283-290.213. Xu, S., Kangwanpong, D., Seielstad, M., Srikummool, M.,Kampuansai, J., Jin, L. and The HUGO Pan-Asian SNPConsortium. (2010). Genetic evidence supports linguisticaffinity of Mlabri - a hunter-gatherer group in Thailand.BMC Genetics, 11(1), 18.214. Yukphan, P., Malimas, T., Lundaa, T., Muramatsu, Y.,Takahashi, M., Kaneyasu, M., Tanasupawat, S., Nakagawa,Y.,Suzuki, K.-I.,Tanticharoen, M. and Yamada, Y. (2010).Gluconobacter wancherniae, sp. nov., an acetic acidbacterium from Thai isolates in the α-Proteobacteria.Journal of General and Applied Microbiology, 56(1), 67-73.215. Yukphan, P., Malimas, T., Muramatsu, Y., Takahashi, M.,Kaneyasu, M., Potacharoen, W., Tanasupawat, S., Nakagawa,Y., Hamana, K., Tahara, Y., Suzuki, K., Tanticharoen, M.and, Yamada, Y. (2009). 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82Annual Report 2010LIST OF PATENTS AND PETTY PATENTSList of Patent / Petty Patent ApplicationsTitle Filing Date Ref. No.Xanthone analogues with anti-herpes simplex activity 16 October 2009 0903001233(Petty patent)Xanthones with anti-herpes simplex activity 16 October 2009 0901004650Calculation of marbling fat for meat quality determination by using random 13 November 2009 0901005076image processing techniqueBiological process for degradation of histamine in high salt containing foods 20 November 2009 0901005189by using halophilic archaea producing histamine dehydrogenaseMethod for production of monoclonal antibodies having desired isotypes 18 December 2009 0901005713The use of yeast cell with phytase on cell surface in combination with apolysaccharide-degrading enzyme to improve nutrient in animal feed15 January 2010 1003000039(Petty patent)DNA vector for expressing gene responsible for the production of proteinand metabolite in filamentous fungi22 January 2010 1003000062(Petty patent)Immunoassay methods for detecting bacterial fruit blotch in cucurbits using 19 February 2010 1001000265monoclonal antibody specific to Acidovorax avenae subsp. citrulliHemoglobin E (HbE) tube kit 26 February 2010 1001000304Recombinant plasmid for the co-production and secretion of at least 2 25 March 2010 1003000265enzymes and production thereof(Petty patent)Pyrone compound that can inhibit the growth of cancer cell 1 April 2010 1001000566Procedure for peptide inhibitor design 29 April 2010 1001000686A gene expression-based method for serological identificationof autoantibodies to cancer cells29 April 2010 1003000366(Petty patent)Continuous biogas production unit 27 May 2010 1003000451(Petty patent)PCR detection method for infectious hypodermal and haematopoietic necrosis 10 June 2010 1001000856virus (IHHNV) in black tiger shrimp using primer pairs to cover the wholeIHHNV genomeDetermination of bovine sperm quality by measuring the RNA level ofprotamine gene24 June 2010 1003000546(Petty patent)Detection of porcine growth rate associated SNPs by using allele specificPCR24 June 2010 1003000547(Petty patent)Detection of porcine production performance and litter size associated SNPsby using allele-specific PCR24 June 2010 1003000548(Petty patent)Detection of porcine meat tenderness and meat quality associated SNPs byusing allele-specific PCR24 June 2010 1003000549(Petty patent)Detection of porcine growth rate associated SNPs 24 June 2010 1003000550(Petty patent)Detection of porcine production performance and litter size associated SNPs 24 June 2010 1003000551(Petty patent)Detection of porcine meat tenderness and meat quality associated SNPs 24 June 2010 1003000552(Petty patent)Unless otherwise specified, all titles presented in the table are for patents filed in Thailand.

National Center for Genetic Engineering and Biotechnology (BIOTEC) 83Title Filing Date Ref. No.Detection of gene defect associated with canine drug allergy 24 June 2010 1003000553(Petty patent)Detection of bovine marbling fat and meat tenderness associated SNPs byusing loop-mediated isothermal DNA amplification24 June 2010 1003000554(Petty patent)Method for lactic acid bacteria identification using DNA microarray 28 July 2010 1001001146Sucrose measuring device based on pulse amperometry for sugar millfactory2 August 2010 1003000692(Petty patent)Simultaneously serotyping of dengue virus by using anti-dengue NS1monoclonal antibodies11 August 2010 1003000726(Petty patent)Methods to enhance expression of influenza A virus neuraminidase in 20 August 2010 1001001267mammalian cells using influenza non-structural protein 1A rapid dual-labelling bioassay for screening of anti-anthracnose agents 2 September 2010 1003000806(Petty patent)Construction of a red fluorescent Colletotrichum capsici recombinant strainand its application2 September 2010 1003000807(Petty patent)Sandworm harvesting device 16 September 2010 1003000884(Petty patent)Protocol for clonal propagation of oil palm plantlets (Elaeis guineensis Jacq.) 27 September 2010 1001001479An effective protocol for rice (Oryza sativa L.) flowering and seed set in 30 September 2010 1001001519plant tissue culture systemDNA plasmid for the production of Vegetative insecticidal proteins (Vips) inBacillus spp. hosts and its application.30 September 2010 1001001520List of Granted Patents / Petty PatentsTitle Granting Date (Petty) Patent No.Determination of risk of developing dengue hemorrhagic fever/dengue shocksyndrome, methods and compositions therefor8 December 2009 US7629117(USA)TB detection kit using one-tube nested PCR technique 8 January 2010 TH27265Nucleic acids that enhance the synthesis of 2-acetyl-1-pyrroline in plantsand fungi12 January 2010 VN8180(Vietnam)Pollen trapping device 22 April 2010 TH5341(Petty patent)Technique for long-term preservation of microbial cells 29 April 2010 TH16673Dengue virus mutant strain, MBU 01-2002 25 May 2010 US7722885(USA)Dried blood spot paper hole punching device for DNA extraction 25 May 2010 TH5396(Petty patent)DNA detection of M. marinum and M. fortuitum complex 1 July 2010 TH28129Identification of Mycobacterium tuberculosis and nontuberculousmycobacteria by one tube multiplex PCR1 July 2010 TH28130Unless otherwise specified, all titles presented in the table are for patents filed in Thailand.

84Annual Report 2010HONORS AND AWARDSProf. Morakot TanticharoenSenior Advisor to the Executive DirectorAPBioChEC Award, presented at the Asia Pacific Biochemical Engineering Conference 2009 (APBioChEC’09) in JapanProf. Watchara KasinrerkBiomedical Technology Research CenterOutstanding Person of the Nation 2009 (Science and Technology), awarded by the National Identity Board, the Prime Minister’sOfficeAssoc. Prof. Dr. Apichart VanavichitRice Gene Discovery UnitFirst runner-up Innovation Award 2009, awarded by Kasetsart UniversityDr. Verawat ChampredaBioresources Technology UnitYoung Scientist Award 2010 for his work on “Enzyme discovery from microbial resources in Thailand and enzyme application inkey local industries”, awarded by the Foundation for the Promotion of Science and Technology under the Patronage of HisMajesty the KingDr. Wonnop VisessanguanBIOTEC Central Research UnitNSTDA Inventor’s Day & Technopreneur Prize 2010 for his work on “Production of pentosanase enzyme from Aspergillus sp.BCC7178 for animal feed”, awarded by the National Science and Technology Development Agency (NSTDA)Dr. Chalermpol KirdmaneeBIOTEC Central Research UnitNSTDA Inventor’s Day & Technopreneur Prize 2010 for his work on “Air filter for air and moisture exchange for tissue culturecontainers”, awarded by the National Science and Technology Development Agency (NSTDA)Dr. Ekachai JenwitheesukGenome InstituteSecond runner-up in The NSTDA Idea to Product Competition 2009: Fast Track to Commercialization for his work on “Canine drugallergy test kit”, awarded by Technology Management Center and Thammasart UniversityDr. Nitsara Karoonuthaisiri et. al.(Dr. Oraprapai Gajanandana, Dr. Plearnpis Luxananil, Dr. Kanyawim Kirtikara, Mr. Ratthaphol Charlermroj and Ms. UmapornUawisetwathana)BIOTEC Central Research UnitNRCT Invention Award 2010 for the development of “Antibody array for multiple detection of food-borne pathogens”, awarded bythe National Research Council of Thailand (NRCT)Dr. Philip ShawBIOTEC Central Research UnitGrand Challenges Explorations Grant 2009 for a research project titled “A new tool for anti-malarial target gene validation”, awardedby the Bill & Melinda Gates FoundationThe Great Gigabase for a research project titled “Comprehensive Transcript Mapping of the 5’ ends of P. falciparum mRNA”,awarded by Roche Diagnostics (Thailand) Ltd., Biogenomed Co., Ltd. and Genome Institute.Dr. Nitsara KaroonuthaisiriBIOTEC Central Research UnitL’Oreal Thailand For Women in Science Fellowship 2009 (Life Science) for her work on “Application of Microarray Technology forResearch and Development in Thailand”, awarded by L’Oreal Thailand and the Thai National Commission for UNESCO

National Center for Genetic Engineering and Biotechnology (BIOTEC) 85Dr. Chawanee ThongpanchangBioresources Technology UnitCST – Wiley Outstanding Publication Award 2009 for her work on “Immobilization of malarial (Plasmodium falciparum) dihydrofolatereductase for the selection of tight-binding inhibitors from combinatorial library”, awarded by the Chemistry Society of ThailandDr. Nipa ChokesajjawateeBIOTEC Central Research UnitEndeavour Executive Awards 2010 for her work on “Principle and advanced application of predictive modeling for construction ofquantitative risk assessment model of Salmonella sp. in chicken”, awarded by the Australian GovernmentDr. Alongkorn AmnuaykanjanasinBioresources Technology Unit2010 IFS Grant for a research project titled “Determination of potential role of reducing clade III PKS gene in theentomopathogenic fungus Beauveria bassiana on pathogenesis against insect pests”, awarded by the International Foundationfor Science (IFS)Dr. Virak VisudtipholeBIOTEC Central Research Unit2010 IFS Grant for a research project titled “Characterization of genes and proteins related to Ca2+ homeostasis and stress in theblack tiger shrimp (Penaeus monodon)”, awarded by the International Foundation for Science (IFS)Dr. Vanvimon SaksmerpromeCenter of Excellence for Shrimp Molecular Biology and Biotechnology2010 IFS Grant for a research project titled “Production and delivery of double-stranded RNA to control yellow-head virus diseasein shrimp”, awarded by the International Foundation for Science (IFS)Best Oral Presentation 2009 for her work on “A novel and inexpensive application of RNAi technology to protect shrimp from viraldisease”, presented at the Meeting of Young Scientists and TRF Senior Research Scholars, organized by Thailand ResearchFund (TRF)Dr. Sithichoke Tangphatsornruang, et al.(Ms. Duangjai Sangsrakru, Ms. Juntima Chanprasert, Ms. Thippawan Yoocha, Ms. Pichahpuk Uthaipaisanwong, Ms. Nukoon Jomchaiand Dr. Somvong Tragoonrung)Genome InstitutePaper titled “The chloroplast genome sequence of mungbean (Vigna radiata) determined by high-throughput pyrosequencing:structural organization and phylogenetic relationships” was highlighted on A-IMBN Research, selected by the Asia-PacificInternational Molecular Biology Network (A-IMBN)Dr. Piyanun Harnpicharnchai, et. al.(Ms. Warasirin Sornlake, Mr. Kittapong Sae-Tang, Dr. Lily Eurwilaichitr and Dr. Sutipa Tanapongpipat)Bioresources Technology UnitPaper titled “Cell-surface phytase on Pichia pastoris cell wall offers great potential as a feed supplement” was highlighted onA-IMBN Research, selected by the Asia-Pacific International Molecular Biology Network (A-IMBN)Mr. Ukrit Rattanachomsri, et. al.(Dr. Sutipa Tanapongpipat, Dr. Lily Eurwilaichitr and Dr. Verawat Champreda)Bioresources Technology UnitJBB Excellent Paper Award for Overseas Researchers 2010 for the paper titled “Simultaneous non-thermal saccharification of cassavapulp by multi-enzyme activity and ethanol fermentation by Candida tropicalis”, awarded by The Society for Biotechnology,JapanMr. Asawin Wanitchang and Dr. Anan Jongkaewwattana, et. al.(Research team of Virology and Cell Technology Laboratory)BIOTEC Central Research UnitOutstanding Poster Presentation Award on “Compatibility of strain-specific amino acids of PA of pandemic H1N1 influenza A virusinfluences its polymerase function and virus growth”, presented at Thailand Conference on Emerging Infectious and NeglectedDiseases (EID2010)

86Annual Report 2010Ms. Pinpunya RiangrungrojBIOTEC Central Research UnitBest Poster Award 2010 (Third Prize) on “Biochemical characterization of adenosine deaminase from Plasmodium falciparum”,presented at The 5 th Annual Symposium of Protein Society of ThailandDr. Jittima PiriyapongsaGenome InstituteThesis Award 2009 (Agricultural Science and Biology) for the PhD thesis titled “Origin and evolution of eukaryotic gene sequencesderived from transposable elements”, awarded by the National Research Council of Thailand (NRCT)Dr. Sumarin SoonsangaBioresources Technology UnitThesis Award 2009 (Agricultural Science and Biology) for the PhD thesis titled “Sensing of organic hydroperoxide by Bacillus subtilisOhrR”, awarded by the National Research Council of Thailand (NRCT)Dr. Nanchaya WanasenBIOTEC Central Research UnitThesis Award 2009 (Medical Science) for the PhD thesis titled “Immuopathogenic mechanisms of non – healing – cutaneousleishmaniasis caused by Leishmania amazonensis”, awarded by the National Research Council of Thailand (NRCT)Mr. Soonthorn Todam and Bala Conservation Youth GroupPeat Swamp and Hala-Bala Rain Forest Research Unit and Nikompattana 10 School, Narathiwat ProvinceGreen Globe Award 2009 (Youth Category) for the work on “Conservation and Integration of Biodiversity in Saiburi River intoSchool Curriculum”, awarded by PTT Public Company Limited

National Center for Genetic Engineering and Biotechnology (BIOTEC) 87Executives and Management TeamExecutive BoardAdvisorsProf. Naksitte CoovattanachaiProf. Sanit AksornkoaeDr. Sermpol RatasukDr. Sakarindr BhumiratanaChairmanProf. Sujin JinahyonVice ChairmanDr. Thaweesak KoanantakoolMembersProf. Narongsak ChaiyabutrProf. Prapon WilairatMr. Pornsil PatchrintanakulProf. Peerasak SrinivesDr. Vichai ChokevivatMrs. Vipajaree PutthamilinprateepProf. Sawasd TantaratanaMr. Somchai CharnnarongkulDr. Somsak ChunharasProf. Amaret BhumiratanaDr. Kanyawim KirtikaraMs. Dussadee SiamhanDirector of the Royal Golden Jubilee PhD Program, Thailand Research Fund (TRF)President, Thailand Environment Institute (TEI)Expert, TEAM Consulting Engineering and Management Co., Ltd. (TEAM)President, King Mongkut’s University of Technology ThonburiPresident, Naresuan UniversityPresident, National Science and Technology Development Agency (NSTDA)Faculty of Veterinary Science, Chulalongkorn UniversityFaculty of Science, Mahidol UniversityVice President, Charoen Pokphand GroupDeputy Secretary General, Thai Chamber of CommerceFaculty of Agriculture, Kasetsart UniversityDirector, Institute for Development of Human Research ProtectionBureau of the BudgetDirector, Thailand Research Fund (TRF)Director General, Department of AgricultureSecretary General, National Health FoundationFaculty of Science, Mahidol UniversityExecutive Director, BIOTECDeputy Executive Director, BIOTECInternational Advisory BoardChairmanProf. Ken-ichi AraiMembersDr. Jill ConleyProf. Mauro GiaccaProf. Paul GreenfieldDr. Ming-Chu HsuProf. Lene LangeProf. Anthony TurnerProf. Dyann WirthProf. Albert Cheung Hoi YuProfessor Emeritus, The University of Tokyo, JAPANDirector, International and Precollege Science Education ProgramsHoward Hughes Medical Institute (HHMI), USADirector, International Center for Genetic Engineering and Biotechnology (ICGEB)Trieste, ITALYVice-Chancellor, The University of Queensland, AUSTRALIAChairman & CEO, TaiGen Biotechnology Co., Ltd., TAIWANVice Dean for Faculty of Engineering, Science and Medicine,Aalborg University, DENMARKCommercial Director & Distinguished Professor of Biotechnology,Cranfield Health, Cranfield University, UKChair, Department of Immunology and Infectious Diseases,Harvard School of Public Health, USAVice-Director and Professor, Neuroscience Research Institute &Department of Neurobiology, Peking University, CHINAAs of 30 September 2010

88Annual Report 2010Management TeamDr. Kanyawim KirtikaraExecutive DirectorMs. Dussadee SiamhanDeputy Executive DirectorDr. Suvit TiaDeputy Executive DirectorMs. Kruawan PotisombatAssistant Executive DirectorDr. Omjai SaimekAssistant Executive DirectorAs of 30 September 2010