Annual Report 2011 National Center for Genetic Engineering and Biotechnology (BIOTEC)
This report was prepared according to the 2011 fiscal year of the Royal Thai Government, from 1 October 2010 – 30 September 2011. Annual Report 2011 National Center for Genetic Engineering and Biotechnology (BIOTEC)
ISBN 978-616-12-0197-5 First Edition January 2012 Number of copies printed 400
Copyright © 2012 by National Center for Genetic Engineering and Biotechnology (BIOTEC) National Science and Technology Development Agency (NSTDA)
Cover images
Thanyasirin: a jasmine glutinous rice (RD6) with blast resistance
Ophiocordyceps halabalaensis: a new species of a fungal pathogen on ant discovered in Hala-Bala Wild Sanctuary, Narathiwat Province
Exploring microbial biodiversity in tropical peat swamp forest using metagenomic approach
From malaria to sleeping sickness: DHFR as drug targets
Published by National Center for Genetic Engineering and Biotechnology (BIOTEC) National Science and Technology Development Agency (NSTDA) 113 Thailand Science Park Phahonyothin Road Klong 1, Klong Luang Pathumthani 12120 THAILAND Tel: +66 (0) 2564 6700 Fax: +66 (0) 2564 6701-5 Website: http://www.biotec.or.th Contents
4 Message from the BIOTEC Executive Director 6 Facts and Figures 14 Research and Development 42 Commercialization and Private Sector Partnership 50 Human Resources Development 56 Public Awareness 62 International Collaboration 70 Impact of BIOTEC’s Output 77 Appendices
77 List of Publications 90 List of Patents and Petty Patents 92 Honors and Awards 95 Executives and Management Team 4
Message from the BIOTEC Executive Director
ur 2011 fiscal year started off with the establishment of three research units in October 2010: Agricultural Biotechnology, OFood Biotechnology and Medical Molecular Biology Research. From this, we will be able to improve the focus of our research as well as establish clear goals.
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In December 2010, BIOTEC was honored by H.R.H. Princess Regarding human resources development, BIOTEC has long Maha Chakri Sirindhorn who bestowed the name “Thanyasirin” been positioning itself to serve as a regional R&D and training to a new blast-resistant glutinous rice, jointly developed by hub in biotechnology by acting as a host to students and Kasetsart University, the Thai Government’s Rice Department, young scientists from abroad to conduct internships and Rajamangala University of Technology Lanna Lampang and early research career development under supervision of BIOTEC. Four years were required to develop Thanyasirin, BIOTEC scientists. This year, BIOTEC was entrusted by Soon (2002 to 2006), using marker-assisted breeding techniques Chun Hyang University (SCH) in Korea to implement a 3-week to achieve the new rice with grain and cooking quality equal training course for twenty undergraduate students to gain to that of RD6, the widely eaten glutinous jasmine rice, with understanding of biotechnology research and development the additional ability to resist rice blast disease. RD6, popularly in the context of Thailand and to experience hands-on grown by farmers in rain-fed lowlands, is highly susceptible to laboratory practice. Throughout the year, we had over one a fungus causing blast disease. This new blast-resistant rice will hundred foreign students and researchers in total from 18 hopefully not only improve rice productivity, but also reduce different countries. the need for fungicides. As this report was being prepared in October/November In April 2011, an effort to integrate databases on biodiversity (2011), Thailand is facing the most devastating flood for the among major institutes within the Ministry of Science and past 70 years. Due to heavier than average rainfall, particularly Technology (BIOTEC, Thailand Institute of Scientific and in September, the campus of Thailand Science Park had to be Technological Research [TISTR] and National Science Museum closed for over 6 weeks, during which the mainline electricity [NSM]), was launched. While BIOTEC’s collection is on special was shut down for two full weeks, posing a challenge to maintain microorganisms such as fungi, actinomycetes and yeasts, TISTR all biological specimens and major scientific equipment. With houses a large collection of algae and plant samples, whereas swift measures put in place, all cultures and biomaterials NSM Natural History Museum has two major collections on preserved at BIOTEC Culture Collection, one of core culture plant and animal reference collections and holotypes (the first collections of the nation, were transferred out of the facility in specimen of a species). The integration of these biodiversity time. We aim to increase our efforts in early 2012 to restore our data will result in a substantial collection of over 100,000 research activities back to schedule. In 2012, we also anticipate specimens. This database has the potential to serve as support newly-initiated projects in response to problems emerging for, in addition to research and development, community- from the floods, such as water quality and water treatment, based ecotourism and prioritization of areas of high diversity production and distribution of rice grain to farmers due to lost for increased government protection and management. crops, for instance – to reiterate our goal to use our expertise in biotechnology to work on the nation’s problems. On the translational side, this year we witnessed a significant step in dengue vaccine development. BIOTEC/NSTDA, Chiang Mai University, Mahidol University jointly licensed dengue vaccine candidates to BioNet-Asia Co., Ltd. for further development into a commercial product. This breakthrough stemmed from a long-term collaborative effort among researchers from these three organizations since 2000. Dr. Kanyawim Kirtikara Executive Director, BIOTEC
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Facts and Figures
IOTEC was first set up under the Ministry for Science, As a premier research institute in Thailand and Asia, BIOTEC BTechnology and Energy on 20 September 1983. After operates research units located at Thailand Science Park the establishment of the National Science and Technology and specialized laboratories hosted by various universities, Development Agency (NSTDA) on 30 December 1991, BIOTEC covering a wide spectrum of research topics from agricultural became one of the NSTDA centers, operating outside the science to biomedical science and environmental science. normal framework of civil service and state enterprises. This In addition to research units, development units have been enabled the Center to operate more effectively to support and established for activities with high commercial potential. These transfer technology for the development of industry, agriculture, are full scale business and production operations designed natural resources, environment and consequently the social and to demonstrate the commercial viability of technologies to economic well-being of Thai people. Other centers under the prospective investors. NSTDA family include National Metal and Materials Technology Center (MTEC), National Electronics and Computer Technology Apart from research and commercialization, BIOTEC activities Center (NECTEC), National Nanotechnology Center (NANOTEC) also include policy research, an outreach program, human and Technology Management Center (TMC). resource development and international relations.
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Organization
Business and Research Capacity Building Administration Technology Transfer
• Research Units located • Development Units • Technical Training Unit • Planning Division at Thailand Science Park • Biotechnology Business • Human Resources and • Evaluation and • BIOTEC’s Satellite Units Development Division Platform Technology Monitoring Division located at universities • Rural Development Development • Budgeting Section and other government Technology Service Unit • Management departments Information System • Research Unit Division Coordination Division • Public Relations Section • Policy Study and • International Biosafety Unit Collaboration Division • Human Resources and Organization Development Section • Building and Maintenance Management Section • Safety, Health and Environment Section • Procurement Section • General Administration Section
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Research Units at Thailand Science Park
Medical Molecular Biology Research Unit • Protein-Ligand Engineering and Molecular Biology Laboratory • Tuberculosis Research Laboratory
Food Biotechnology Research Unit • Food Biotechnology Laboratory Agricultural Biotechnology Research Unit • Monoclonal Antibody Production Laboratory • Animal Physiology Laboratory • Aquatic Molecular Genetics and Biotechnology Laboratory • Shrimp-Virus Interaction Laboratory • Microarray Laboratory • Starch Biosynthesis Laboratory • Plant Physiology and Biochemistry Laboratory • Plant Molecular Genetics Laboratory • Plant Research Laboratory • Virology and Cell Technology Laboratory
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Genome Institute • Genomic Research Laboratory • Proteomics Research Laboratory • Biostatistics and Bioinformatics Laboratory
Bioresources Technology Unit • BIOTEC Culture Collection Laboratory • Biotechnology Law Agricultural Biotechnology Research Unit • Bioresources Research Laboratory • Monoclonal Antibody Production Laboratory • Bioassay Laboratory • Animal Physiology Laboratory • Enzyme Technology Laboratory • Aquatic Molecular Genetics and Biotechnology Laboratory • Microbial Cell Factory Laboratory • Shrimp-Virus Interaction Laboratory • Fermentation Technology and Biochemical Engineering Laboratory • Microarray Laboratory • Microbial Engineering Laboratory • Starch Biosynthesis Laboratory • Mycology Laboratory • Plant Physiology and Biochemistry Laboratory • Information Systems Laboratory • Plant Molecular Genetics Laboratory • Geoinformatics Laboratory • Plant Research Laboratory • Ecology Laboratory • Virology and Cell Technology Laboratory
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Research Units at 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 Province
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Development Units
• Shrimp Biotechnology Business Unit (SBBU) • Dairy Cattle Research and Business Development Project • Shrimp Genetic Improvement Center (SGIC) • Nuclear Polyhedrosis Virus Pilot Plant for Insect Pest Control
Finance
For 2011 fiscal year, BIOTEC’s budget totaled 753 million Baht and expenditure was 643.73 million Baht. An income of 81 million Baht was earned from external sources outside NSTDA.
Expenditure of 644 MB Research Expenditure, 483 MB 75% HRD in Biotechnology, 23 MB 3% Technology Transfer, 18 MB 3% General Administration, 45 MB 7% ICT Support & Services, 36 MB 6% Building Operation & Services, 39 MB 6%
Income of 81 MB Research Fund, 34 MB 42% Contract and Collaborative Research, 22 MB 28% Licensing Activity, 1 MB 2% Technical Services, 20 MB 24% Workshop/Conference, 3 MB 3% Others, 1 MB 1%
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Human Resources
As of 30 September 2011, BIOTEC employed a total of 570 staff.
BIOTEC Staff by Education 29% PhD, 163 36% M.Sc., 204 30% B.Sc., 172 15% Below B.S., 31
BIOTEC Staff by Job Function 3% Executive, 17 14%
Administrative, 81 83% R&D and Scientific Staff, 472
Publications
BIOTEC produced 218 publications, with 24 published in journals with impact factors greater than 4. Two papers were published in journals with impact factors greater than 10. Sixteen were published in non-citation index journals.
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Patents
BIOTEC applied for twenty-two patents, one of which was applied in the USA, and fourteen petty patents. Four patents and five petty patents were obtained. A patent on “BADH2 nucleic acids associated with grain aroma” was granted in the USA, whereas “Nucleic acids that enhance the synthesis of 2-acetyl-1-pyrroline in plants and fungi” in Japan.
Honors and Awards
BIOTEC researchers and affiliated staff were presented with 35 awards, both locally and internationally.
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RESEARCH AND DEVELOPMENT
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IOTEC R&D and technical program covers a wide spectrum of Bresearch topics from agricultural to medical sciences and from energy to environmental sciences. Plant biotechnology focuses on three economically important plants: rice, cassava, and oil palm. Animal biotechnology at BIOTEC focuses on shrimp and dairy cow, whereas food biotechnology aims to improve and upgrade the processing and quality of fermented food, with topics such as food safety and risk assessment, food chemistry, and starter culture technology. Medical biotechnology focuses on tropical as well as emerging diseases such as malaria, tuberculosis, dengue fever and influenza. On environment, BIOTEC emphasizes on the study of the rich microbial diversity, the preservation, utilization, and conservation of bioresources. Biogas and other renewable energy is the topic of focus in energy research theme.
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Highlights on Rice Biotechnology
Molecular breeding for rainfed lowland rice produced. Some improved lines appeared to have traits in The research team from the Rice Gene Discovery Unit (RGDU), a combination such as tolerance to submergence and bacterial satellite research unit of BIOTEC in collaboration with Kasetsart blight or submergence and resistance to brown planthopper, University, developed genomic tools for discovering genes as well as some with three traits combined together such as underlying important economic traits such as resistance to tolerance to submergence and resistance to bacterial blight and environmental stresses (salt tolerance, flooding or drought), brown planthopper. Lines are being developed and selected disease resistance (blast and bacterial leaf blight) and pest by MAS at RGDU such as Chainat 1 with brown planthopper (brown plant hopper, white backed plant hopper) by studying resistance, RD6 with several resistant QTL pyramided against the biodiversity of rice varieties and finding molecular markers blast disease. Improved lines of KDML105 and RD6 with brown for the Thai rice breeding program. In a collaborative project planthopper resistance were validated in the greenhouse at between RGDU, BIOTEC and the Rice Department, resistance RGDU. At Ubon Ratchathani Rice Research Center of the Rice against biotic and abiotic stresses of rainfed lowland rice such Department, KDML105, RD6 and Surin 1 with drought tolerance as Khao Dawk Mali 105 (KDML105) and RD6 (glutinous rice) were validated under field conditions. Farmer’s field trials of were improved with the use of molecular tools. RD6 with blast resistance (cultivar: Thanyasirin), KDML105 with submergence tolerance and resistance to bacterial blight and The main objective of breeding rainfed lowland rice is to brown planthopper were conducted to evaluate performance develop lines by using marker assisted selection to ensure that and adaptation of the promising lines. Finally, promising both KDML105 and RD6 maintain their desirable agronomic lines will be certified by the Rice Department to hasten seed characteristics or to breed lines that improve upon the original production and supply to farmers. In addition, three elite lines KDML105 and RD6. Backcross introgression lines of KDML105 (KDML105 with submergence tolerance, KDML105 with salinity and RD6 with improved resistance to bacterial blight, blast, tolerance and RD6 with blast resistance) are in the process of brown planthopper, flooding, salinity and drought were varietal registration.
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Ideotype breeding by marker-assisted Delivery of new rice varieties to farmers selection Homcholasit is an irrigated rice cultivar (non photoperiod Marker-assisted selection (MAS) is an efficient method for sensitive) with aroma and good eating quality. It is derived from hastening line development. Its efficiency in transferring a cross between KDML105 and IR57514 and was developed gene/QTL conferring traits related to biotic and abiotic for flash flooding tolerance. It can survive under water for up stresses and grain qualities has been proven in rice by many to 2 to 3 weeks. Homcholasit can be planted all throughout breeding programs around the world. A research team from the year, providing yield of 5-6 tons per hectare. Towards the RGDU, in cooperation with the Phitsanulok Rice Research end of the year 2010, around August to December, BIOTEC Center, developed the desirable lines with submergence delivered 400 kg of Homcholasit seeds to farmers in Phak Hai tolerance and jasmine-like cooking quality for rice varieties District, Ayuthaya Province, through agricultural cooperatives. grown in rainfed and irrigated lowland ecosystems in the Homcholasit was planted and yielded about 5 tons/hectare Mekong region of Southeast Asia by using this technology. in the non-flooded area while in areas flooded for 12 days at Hybridization between varieties IR57514 and Kao Dawk Mali the flowering stage, Homcholasit survived and yielded about 105 (KDML105) was initiated with the goal of producing an 1 ton/hectare. In addition, this cultivar was also delivered to ideotype that combines submergence tolerance and jasmine- 3 other provinces namely Chainat, Ang Thong and Uttaradit. like cooking quality. Through the single seed descent (SSD) During crop season of 2010 to 2011, total yield of this cultivar method, a large population of recombinant inbred lines was 451 tons from 154 hectares. (RILs) was developed. MAS was used to identify the ideotype of the offspring carrying the favored alleles of the Sub1, Wx, Thanyasirin, the name bestowed by H.R.H. Princess Maha Chakri badh2 and SSIIa loci, respectively. Sixty six lines were selected. Sirindhorn, was developed under collaboration among RGDU, All lines exhibited submergence tolerance and jasmine-like Kasetsart University, Rajamangala University of Technology cooking quality and displayed low amylose content, fragrance Lanna and the Rice Department. Thanyasirin is a jasmine and a high alkali spreading value. A wide range of agronomic glutinous rice (RD6), with blast resistance added into its characteristics was observed and some of the lines were genome using molecular markers in the backcross breeding superior in yield component, as compared to their parents. process. Thanyasirin has good cooking and eating qualities. It In this study, the superior genotype of Homcholasit was shows drastic improvement in leaf and neck blast resistance discovered. It has desirable traits of submergence tolerance, of a broad range of spectrum in the rainfed lowland areas. low amylase content, fragrance, lodging resistance (short plant Thanyasirin seeds were delivered to farmers in 5 provinces such height), is non-photoperiod sensitive and high yielding (7.2 t/ as Nan, Chiang Rai, Lampang, Chiang Mai and Chaiyaphum. ha in the farmer’s field). This work was published inField Crop Research in June 2011.
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Highlights on Cassava Research
SSR and EST-SSR-based genetic linkage map encompassed around 88% of the cassava genome indicating of cassava that the map was almost complete. Therefore, sufficient markers now exist to encompass most of the genomes and efficiently Simple sequence repeat (SSR) markers provide a powerful map traits in cassava. This work was published in Theoretical tool for genetic linkage map construction that can be applied and Applied Genetics in April 2011. for identification of quantitative trait loci (QTL). The research team at the Genome Institute in collaboration with Mahidol Genome scan for quantitative trait loci University and the Ministry of Agriculture and Cooperatives, developed a total of 640 new SSR markers from an enriched affecting cyanogenic potential of cassava genomic DNA library of the cassava variety ‘Huay Bong 60’ and root 1,500 novel expressed sequence tag-simple sequence repeat Cassava (Manihot esculenta Crantz) can produce cyanide, (EST-SSR) loci were developed from the GenBank database. a toxic compound, without self-injury. Researchers at the To construct a genetic linkage map of cassava, a 100 F1 line Genome Institute, in collaboration with Mahidol University, mapping population was developed from the cross Huay the Ministry of Agriculture and Cooperatives and the Southern Bong 60 by ‘Hanatee’. Polymorphism screening between the Illinois University (USA), identified quantitative trait loci parental lines revealed that 199 SSRs and 168 EST-SSRs were (QTL) associated with the cyanogenic potential (CN) in an identified as novel polymorphic markers. Combined with outbred population derived from ‘Hanatee’ x ‘Huay Bong previously developed SSRs, the team constructed a linkage 60’, two contrasting cultivars. CN was evaluated in 2008 map consisting of 510 markers encompassing 1,420.3 cM and and in 2009 in Rayong Province, and in 2009 in Lopburi distributed on 23 linkage groups with a mean distance between Province, Thailand. QTL analysis affecting CN was performed markers of 4.54 cM. Comparison analysis of the SSR order on with 303 SSR markers. Five new QTL affecting CN were the cassava linkage map and the cassava genome sequences successfully identified from 4 linkage groups: CN08R1 provided the location of 284 scaffolds on the genetic map. from 2008 at Rayong, CN09R1 and CN09R2 from 2009 at Although the number of linkage groups reported here revealed Rayong, and CN09L1 and CN09L2 from 2009 at Lopburi. that this F1 genetic linkage map is not yet a saturated map, it Discovery of these QTL can provide useful markers to assist in
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cassava breeding and studying genes affecting the trait. For leaf proteins, which can help understand how biochemical molecular breeding, QTL analysis was often used to identify processes in cassava leaves may be involved in storage root trait-linked markers in order to facilitate marker-assisted development. This study involved collaboration between the selection (MAS). QTL identification was used not only to Rayong Field Crop Research Center, Mahidol University and assist the breeding program, but also to gain understanding Proteomic Research Laboratory of the Genome Institute. The of the loci and underlying genes and their effects. This work progress achieved in this research was published in Planta in was published in BMC Genomics in May 2011. June 2011.
Leaf proteomic analysis in cassava during Electrospun polylactic acid and cassava plant development starch fiber by conjugated solvent Tuberization in cassava (Manihot esculenta Crantz) occurs technique simultaneously with plant development, suggesting Electrospinning is a widely used technique for making competition of photoassimilate partitioning between the continuous fiber with diameter ranging from tens nanometer shoot and the root organs. In potato, which is the most widely to tens micron. Electrospun fibers are created from polymer studied tuber crop, there is ample evidence suggesting that solution by using electrical force as a mean for fiber fabrication. metabolism and regulatory processes in the leaf may have Researchers from the Cassava and Starch Technology Research an impact on tuber formation. To search for leaf proteins Unit, a joint research unit between BIOTEC and Kasetsart putatively involved in regulating tuber generation and/or University, have created electrospun polylactic acid (PLA) and development in cassava, comparative proteomic approaches cassava starch fiber using the conjugated solvent technique. have been applied to monitor differentially expressed leaf By introducing a conjugated solvent during mixed solution proteins during root transition from fibrous to tuberous. preparation, it is possible to create a homogeneous and Stringent cross comparison and statistical analysis between spinnable mixed solution. Unlike emulsifier, conjugated solvent two groups with different plant ages revealed a number of evaporates during fiber fabrication without any residual traces. protein spots whose abundance were significantly altered Conjugated solvent is unable to dissolve PLA and cassava starch, during week 4 to week 8 of growth. Of these, 39 spots were thus exhibiting a major difference when compared to the co- successfully identified by ion trap LC–MS/MS. The proteins span solvent system which is used for dissolving multiple solutes. various functional categories from antioxidant and defense, Composite fiber from PLA and cassava starch has potential in carbohydrate metabolism, cyanogenesis, energy metabolism, medical applications for making electrospun tissue scaffolding, miscellaneous and unknown proteins. Results suggested since raw materials are made of biodegradable polymer. This possible metabolic switches in the leaf that may trigger/regulate work was reported in Materials Letters and a patent was filed storage root initiation and growth. This study provides a basis in December 2010. for further functional characterization of differentially expressed
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Highlights on Shrimp Biotechnology
Bacterial community associated with the Molecular cloning and expression analysis intestinal tract of P. monodon in commercial of the MAPK1 gene and protein during farms ovarian development of P. monodon Knowledge of the intestinal bacterial community of the black Isolation and characterization of genes and/or proteins tiger shrimp is important for maintaining sustainable aquaculture differentially expressed in ovaries are necessary for production. The research team at the Aquatic Molecular Genetics understanding ovarian development in the giant tiger shrimp and Biotechnology Laboratory and the Microarray Laboratory, (Penaeus monodon). The research team at the Aquatic Molecular under the Agriculture Biotechnology Research Unit, examined Genetics and Biotechnology Laboratory of the Agricultural the intestinal bacterial community associated with juvenile black Biotechnology Research Unit, characterized the full-length tiger shrimp during the grow-out stage in different commercial cDNA of P. monodon mitogen-activating protein kinase 1 farms in eastern Thailand using 16S rDNA PCR-DGGE and 16S (PmMAPK1). PmMAPK1 was more abundantly expressed in rDNA clone library methods. Bacteria in the γ-Proteobacteria class ovaries than in testes of P. monodon and differential expression were the only common bacteria group found in the intestinal levels of PmMAPK1 mRNA during ovarian development of tracts of shrimp from all farms. The dominant bacterial genera in intact broodstock, where it peaked in stage III ovaries and the intestinal population of each shrimp varied among different slightly decreased afterwards. Likewise, the expression level farms, and these genera were Vibrio, Photobacterium, Aeromonas, of PmMAPK1 in stage IV ovaries was significantly greater than or Propionigenium (phylum Fusobacteria). Additionally, strictly that in stages I, II ovaries of eyestalk-ablated broodstock. The anaerobic bacteria such as Propionigenium and Fusibacter were PmMAPK1 transcript was localized in ooplasm of previtellogenic found. These intestinal bacterial communities varied significantly oocytes. In intact broodstock, the expression of the PmMAPK1 among different commercial farms and were distinct from their protein was clearly increased from previtellogenic ovaries in rearing water. The results provide descriptive structures of the subsequent stages of ovarian development. In contrast, the intestinal bacterial communities of P. monodon in commercial level of ovarian PmMAPK1 protein was comparable during farms. This work was published on-line in Microbiology Ecology oogenesis in eyestalk-ablated broodstock. The PmMAPK1 in September 2011. protein was localized in ooplasms of previtellogenic and
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vitellogenic oocytes. It was also detected around the nuclear cDNA microarray for identification of genes membrane of early cortical rod oocytes in both intact and relevant to testicular development in P. eyestalk-ablated broodstock. Results indicated that PmMAPK1 monodon gene products seem to play functional roles in the development and maturation of oocytes/ovaries in P. monodon. This work Poor reproductive maturation in captive male broodstock was published in Molecular Reproduction and Development of the black tiger shrimp (Penaeus monodon) is one of the in May 2011. serious problems faced by the aquaculture industry. In this study, a new version of cDNA microarray, UniShrimpChip, was cDNA microarray approach for analyzing constructed from the P. monodon EST libraries of 12 tissues, transcriptional changes in P. monodon containing 5,568 non-redundant cDNA clones from 10,536 unique cDNA in the P. monodon EST database. The overall A cDNA microarray comprised of 9,990 different ESTs obtained gene expression patterns from the microarray experiments from the Penaeus monodon EST project (http://pmonodon. revealed distinct transcriptomic patterns between the wild biotec.or.th) was employed to identify viral (white spot and and domesticated groups. Moreover, differentially expressed yellow head viruses) and bacterial (Vibrio harveyi) responsive genes from the microarray comparisons were identified, and genes in the hemocytes of P. monodon at 6, 24 and 48 hours the expression patterns of eight selected transcripts were post injection (hpi). The number of differentially expressed subsequently confirmed by reverse-transcriptase quantitative genes found was highest in shrimps infected with WSSV PCR (RT-qPCR). Among these, expression levels of six subunits (1954 genes) followed by YHV (1136 genes) and V. harveyi (CSN2, 4, 5, 6, 7a, and 8) of the COP9 signalosome (CSN) gene (420 genes). Changes in shrimp gene expression were highest family in wild and different ages of domesticated brooders at the late infection stage for both viruses, whilst that for V. were examined by qPCR. Among the six subunits, CSN5 and harveyi induced gene expression was mainly found at the CSN6 were most highly expressed in wild brooders and least early infection stage, but the repression of genes was mainly expressed in the 18-month-old domesticated group. The found in the mid stage of infection. Shrimp genes specifically results show that there are obvious differences between the upregulated by each particular pathogen were identified by wild and domesticated shrimp at the transcriptomic level. the research team at the Center of Excellence for Molecular Furthermore, the study suggested that the CSN gene family Biology and Genomics of Shrimp, a cooperation between might have been involved in reproduction and development of BIOTEC and Chulalongkorn University. This work was published this economically important species. This work was published in Fish and Shellfish Immunology in January 2011. in BMC Molecular Biology in April 2011.
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New platform for detection of shrimp Genome fragments of the P. stylirostris viruses by LAMP using a portable multi- densovirus in the giant tiger shrimp channel turbidimeter genome have implications for viral disease Researchers at the Center of Excellence for Shrimp Molecular diagnosis Biology and Biotechnology (Centex Shrimp), a joint Center A research team at Centex Shrimp examined the random of Excellence between BIOTEC and Mahidol University, have genomic insertions of densovirus (also called Infectious acquired expertise in the development of diagnostic probes Hypodermal and Hematopoietic Necrosis Virus, IHHNV) into the and kits based on shrimp industry needs to assist in preventing giant tiger shrimp genome by using PCR analysis with seven the outbreak of disease. In collaboration with the National overlapping IHHNV specific primers combined with genome Electronics and Computer Technology Center (NECTEC), a new walking technique to reveal a viral insert linked to a host platform for detection of shrimp viruses by loop-mediated microsatellite-like fragment. The results revealed probability isothermal amplification (LAMP) using a designed portable of two slightly different inserts on paired chromosomes in multi-channel turbidimeter has been developed by using one specimen and similar insertions could occur in several the Taura Syndrome Virus (TSV) as a model. The detection shrimp specimens, including those infected with IHHNV but sensitivity of turbidity measurement was comparable to showing no signs of disease. Some inserts gave false positive agarose gel electrophoresis detection of RT-LAMP and PCR test results using 309F/R primers and a new IQ2000 test nested RT-PCR (IQ2000™) products. Cross reactions with other protocol currently recommended for detection of infectious shrimp viruses were not found, indicating that the RT-LAMP- IHHNV. This is the first experimental results to support that a turbidity measurement was highly specific to TSV and the random number and length of sequence fragments from a results could be detected within 1 hour. The new alternative single virus genome may occur in the shrimp genome. False platform is portable and can be used for detection not only positive results with the recommended methods for infectious in the laboratory but also in the field. This work filed for a Thai IHHNV associated with inserts, has resulted in negative effects patent in March 2011 and was also published in the Journal on international seafood trade. This work was published in of Virology Methods in August 2011. Viral Research in September 2011.
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Highlights on Food Biotechnology
Immunoassay based on carbon nanotubes- enhanced ELISA for Salmonella The research team at the Biochemical Engineering and Pilot Plant Research and Development Unit, a joint research unit between BIOTEC and King Mongkut’s University of Technology Thonburi, demonstrated a new method to enhance the Biogenic amine formation in Thai fermented sensitivity of enzyme linked immunosorbent assay (ELISA) for sausage and reduction by commercial detection of Salmonella enterica serovar Typhimurium, a serious life threatening pathogen causing human gastroenteritis. starter culture Single-walled carbon nanotubes (SWCNTs) were employed as Biogenic amines are of concern for Nham, a traditional Thai a labeling platform for antibody and horseradish peroxidase fermented pork sausage, due to their toxicological effects (HRP) co-immobilizing, resulting Ab/SWCNTs/HRP bioconjugate on nervous, blood pressure, gastric and intestinal systems. used in the experiments. The combination between Ab and Researchers at the Food Biotechnology Research Unit have HRP on the SWCNTs complement each other in the way that studied the influence of raw pork meat quality and starter one is used as a recognition element and the other as a signal culture inoculation on biogenic amines accumulation in Nham. generator. A large surface area to volume ratio of SWCNTs It was found that the formation of biogenic amines (cadaverine, permits a high amount of enzyme binding and hence signal putrescine, histamine and tyramine) was significantly higher amplification. The results have shown the proof-of-concept in Nham processed from stored meat. Accumulation of these using Ab/SWCNTs/HRP bioconjugate as a detecting molecule biogenic amines in Nham was reduced significantly by the to detect S. enterica serovar Typhimurium through classic ELISA addition of Lactobacillus plantarum BCC 9546, a commercial experiments. By replacing a commercial Ab-HRP with the Ab/ Nham starter culture. The results indicate the importance of SWCNTs/HRP bioconjugate, a 100-times greater detection limit using fresh meat products and the inclusion of an appropriate was achieved when tested in food samples. The simplicity starter culture to minimize the formation of biogenic amines and efficacy of using this bioconjugate in ELISA could help during the process of Nham fermentation, resulting in a safer to escape the sensitivity limitations of the technique itself. product for the consumer. This work was published in Food This work was published in Biosensors and Bioelectronics in Chemistry in May 2011. February 2011.
Annual Report 2011 24 Highlights on Plant and Animal Biotechnology
16 of which are duplicated in the inverted repeat. Of the 112 unique genes, 78 are predicted protein-coding genes, 4 are ribosomal RNA genes and 30 are tRNA genes. Relative to other plant chloroplast genomes, a unique rearrangement in the rubber tree chloroplast genome, a 30-kb inversion between the trnE(UUC)-trnS(GCU) and the trnT(GGU)-trnR(UCU), was observed. A comparison between the rubber tree chloroplast genes and cDNA sequences revealed 51 RNA editing sites in which most (48 sites) were located in 26 protein coding genes and the other sites were in introns. Phylogenetic analysis based on chloroplast genes demonstrated a close relationship between Hevea and Manihot in Euphorbiaceae and provided a strong support for a monophyletic group of the eurosid I. The results of this research were published in Gene in April 2011.
Bovine embryo sex determination by multiplex-LAMP Maintaining an appropriate sex ratio between male/female animals is one of the important aspects for livestock farm management. Though commercially available, test kits for bovine embryo sex determination are costly and usually detect only the Y-determinant without internal control. A Characterization of the complete collaborative research project between the Food Biotechnology Research Unit and the Animal Physiology Laboratory of the chloroplast genome of rubber tree Agricultural Biotechnology Research Unit has been undertaken Rubber tree (Hevea brasiliensis) is an economically important to develop a rapid and cost-effective method for bovine plant and widely grown for natural rubber production. However, embryo sex determination by simultaneous amplification of genomic research of rubber tree has lagged behind other the Y-chromosome and internal control in a single tube using species in the Euphorbiaceae family. A research team at the multiplex loop-mediated amplification (multiplex-LAMP). This Genome Institute performed shotgun genome sequencing method is fast (within 30-60 minutes), simple, and highly of H. brasiliensis using the 454 pyrosequencing technology specific because of the embedded internal amplification and obtained the complete chloroplast genome sequence. control. The result is easy to read by visual inspection and The complete chloroplast genome sequence of rubber tree only simple equipment, such as a temperature control box was reported as being 161,191 bp in length including a pair or a water bath and a UV- lamp are required. The simplicity of inverted repeats of 26,810 bp separated by a small single of the detection method and associated equipment allows copy region of 18,362 bp and a large single copy region of for on-site detection when needed. A patent application of 89,209 bp. The chloroplast genome contains 112 unique genes, this method was made in April 2011.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 25
Highlights on Malaria Research
pyrimethamine-resistant mutation isolated from the field. A transgenic P. berghei line, in which endogenous Pbdhfr allele was replaced with the mutant PfdhfrS108N, was generated and confirmed to have normal growth rate compared to the parental non-transgenic parasite and also able to confer resistance to pyrimethamine. This study demonstrated the power of the transgenic P. berghei system to predict drug-resistant Pfdhfr mutations in an in vivo parasite/host setting. The system could be utilized for identification of possible novel drug-resistant mutants that could arise against new antifolate compounds and for prediction the evolution of resistant mutations. This work was published in Malaria Journal in May 2011.
Trypanosomal dihydrofolate reductase reveals natural antifolate resistance Selection of drug resistant mutants from Dihydrofolate reductase (DHFR) is a potential drug target in folate metabolism for Plasmodium spp. and other parasites. random library of Plasmodium falciparum Trypanosoma brucei, a causative agent of human African sleeping A major problem that hinders malaria control is antimalarial sickness, is naturally resistant to the classical antifolates such as drug resistance which is associated with mutations of the pyrimethamine, cycloguanil, or trimethoprim. Understanding drug target enzymes within the parasites. Prediction of drug of natural antifolate resistance is useful for the design of new resistance mutations could provide some insights into effective drugs that targetT. brucei DHFR (TbDHFR). Researchers the design of effective new drugs against resistant parasites. at BIOTEC Protein-Ligand Engineering and Molecular Biology BIOTEC researchers at the Protein-Ligand Engineering and Laboratory in collaboration with Mahidol and Chulalongkorn Molecular Biology Laboratory of the Medical Molecular Biology universities, have studied natural antifolate resistance of TbDHFR Research Unit, in collaboration with Chiang Mai University, have based on information obtained from co-crystal structure of been engaged in research towards developing antimalarial TbDHFR compared with malarial Plasmodium falciparum DHFR drugs to overcome multi-drug resistant parasites for decades. (PfDHFR). The results revealed the structure of TbDHFR is similar The team has developed an animal Plasmodium parasite model to a mutant PfDHFR, rather than a wild-type enzyme. TbDHFR for identification of possible new Plasmodium falciparum has steric hindrance for rigid inhibitors like pyrimethamine. A dihydrofolate reductase (Pfdhfr) mutants that confer resistance missing residue on TbDHFR active-site loop and the presence to antifolate drugs. The researchers generated libraries of of Ile51 widens its active site further than the structural effect Pfdhfr mutants by using the error-prone PCR technique of Asn51Ile, which is observed in PfDHFR structures. Mutations and then episomally transfected the mutant libraries to the of TbDHFR at Thr86 resulted in 10-fold enhancement or 7-fold Plasmodium berghei parasite. The transfected P. berghei were reduction in the rigid inhibitors affinities for Thr86Ser or selected with antifolate drug pyrimethamine and analyzed Thr86Asn, respectively. The co-crystal structure of TbDHFR with for the pyrimethamine-resistant Pfdhfr mutants. It was found a flexible antifolate WR99210 suggests that its greater affinity that the principal mutation identified from this experiment results from its ability to avoid such Thr86 clash and occupy was the Pfdhfr with mutation at amino acid position 108 the widened binding space similarly to what is observed in from serine (S) to asparagine (N), coincident with the first the PfDHFR structures. The results of this study were published
Annual Report 2011 26
reduced from 16 hours to 4 hours by the use of dynamic hybridization scheme. Dynamic hybridization also increased the average fluorescent signal intensities and the signal- to-noise ratios in both Cy3 and Cy5 channels over a static hybridization, allowing more effective spots to be included in the subsequent microarray data analysis. This provides an immense advantage in transcriptomic profiling studies as the additional good spots detected under dynamic hybridization, but not static hybridization could have potential biological or medical significance in, for example, drug target and disease biomarker discovery studies. This work was published in the Lab on a Chip in February 2011.
Determination of the Plasmodium vivax schizont stage proteome With the genome of the malaria parasite Plasmodium vivax sequenced, it is important to determine the proteomes of the in ACS Chemical Biology in June 2011. The article was selected parasite in order to assist efforts in antigen and drug target and evaluated by Prof. Vivian Cody, a Member of the Faculty discovery. Since a method for continuous culture of P. vivax of 1000 (F1000), and was placed in the F1000 library of the top parasite is not available, a proteome of the erythrocytic stages 2% of published articles in biology and medicine. using fresh parasite isolates from patients was conducted in a joint study between researchers from the Genome Institute, Simple and low-cost platform for dynamic Mahidol University, Armed Forces Research Institute of DNA hybridization Medical Sciences (AFRIMS), Virginia Tech and Pennsylvania BIOTEC researchers at the Protein-Ligand Engineering and State University. In schizont-enriched samples, 316 proteins Molecular Biology Laboratory of the Medical Molecular Biology were confidently identified by tandem mass spectrometry. Research Unit, in collaboration with the National Electronics Almost 50% of the identified proteins were hypothetical, and Computer Technology Center (NECTEC), developed a while other major categories include proteins with binding piezoelectric microagitation platform for enhancing DNA function, protein fate, protein synthesis, metabolism and microarray hybridization. cellular transport. To identify proteins that are recognized by host humoral immunity, parasite proteins were separated Plasmodium falciparum DNA microarrays and total RNA extracted by two-dimensional gel electrophoresis and screened by from parasite cells were used to perform DNA microarray Western blot using an immune serum from a P. vivax patient. experiments. DNA hybridization was performed in a sealed Mass spectrometry analysis of protein spots recognized by chamber containing a DNA microarray glass chip, coupling the serum identified four potential antigens including PV24. medium and piezoelectric transducers. Compared with the The recombinant protein PV24 was recognized by antibodies conventional methods of static hybridization scheme, this new from vivax malaria patients even during the convalescent platform could enhance the efficiency of DNA hybridization by period, indicating that PV24 could elicit long-lasting antibody using piezoelectric microagitation through a liquid coupling responses in P. vivax patients. This work was published in medium which allows ultrasonic wave transmission to the Journal of Proteomics in August 2011. glass chip. Required hybridization time was substantially
National Center for Genetic Engineering and Biotechnology (BIOTEC) 27
Highlights on Dengue Research the first vaccination. This work is a joint effort among various partners, namely the Medical Biotechnology Research Unit, a joint research unit among BIOTEC, Mahidol University and Chiang Mai University, Imperial College London, Hospital for Tropical Disease in Vietnam, Faculty of Medicine Siriraj Hospital, Mahidol University, Khon Kaen Hospital and Songkha Hospital. This work was published in the Journal of Virology in October 2010.
Novel serotyping-NS1-ELISA to identify serotypes of dengue virus Dengue virus (DENV), which causes the mosquito-borne disease dengue hemorrhagic fever (DHF), consists of four serotypes co-circulating in endemic areas. Currently, DENV serotypes can be identified by laborious virus isolation followed by immunofluorescent assay and sophisticated RT-PCR. Researchers from the Medical Biotechnology Research Unit, In-depth analysis of original antigenic sin in in collaboration with the Biomedical Technology Research dengue virus infection Center, have developed a new assay designated as "serotyping- Dengue infection, although frequently mild, can lead to dengue NS1-ELISA" to detect the NS1 protein and to identify DENV hemorrhagic fever (DHF) which can be life threatening. DHF serotypes simultaneously. The monoclonal antibodies (Mabs) is more common in secondary dengue infections, implying a against NS1 of each DENV serotype were produced and applied role for the adaptive immune response in the disease. There is to detect the NS1 antigen, which was previously captured by currently much effort toward the design and implementation a flavivirus cross-reactive anti-NS1 Mab. Serotyping accuracy of a dengue vaccine but these efforts are made more difficult of the developed assay was validated with NS1 from DENV- by the challenge of inducing durable neutralizing immunity to infected cell culture supernatants and from well-characterized all four viruses. Domain 3 of the dengue virus envelope protein clinical specimens. In acute phase plasmas from DENV infected (ED3) has been suggested as one such candidate because it patients, 65 out of 85 specimens were positive to one of the contains neutralizing epitopes and it was originally thought four serotypes in this assay. Among those positive samples, that relatively few cross-reactive antibodies are directed to identification of DENV serotypes by serotyping-NS1-ELISA was this domain. The research team has performed a detailed 100% accurate for DENV1, 3 and 4 and 82.4% for DENV2 as analysis of the anti-ED3 response in a cohort of patients compared with standard RT-PCR. Assay specificity was 100%. suffering either primary or secondary dengue infections The developed serotyping-NS1-ELISA provides an alternative from Thailand. Robust responses of neutralizing antibodies for simultaneous detection of DENV NS1 and identification directed to ED3 domain show dramatic evidence of original of its serotype in acute patient’s specimens. The assay would antigenic sin in secondary infections both in terms of binding be applicable for dengue diagnosis and epidemiological and enhancement activity. This basic knowledge has important studies. This work was published in Journal of Clinical Virology implications for dengue vaccine design because heterologous in February 2011. boosting is likely to maintain the immunological footprint of
Annual Report 2011 28
Highlights on Influenza Research
N terminus of PA polymerase of swine- origin influenza virus H1N1 genes, especially the PA segment, to reassort with those of other circulating strains would be limited or even unlikely in Despite several lines of evidence suggesting possible the absence of specific mutations. This article was published mechanisms by which the influenza virus polymerase complex, in Virus Research in November 2010 and was highlighted on comprising PB2, PB1 and PA, work in concert during virus A-IMBN Research, a website intended to highlight some of the replication, exactly how they function is not entirely understood. best research being published by members of the Asia-Pacific The N terminal region of the PA subunit has been shown International Molecular Biology Network. to play a key role in various functions through a number of conserved amino acid residues. However, little is known about Atypical characteristics of nucleoprotein of the role of amino acids reported to be unique for a virus strain. Researchers at the Virology and Cell Technology Laboratory pandemic influenza virus H1N1 of the Agricultural Biotechnology Research Unit investigated Sequence analysis of the nucleoprotein (NP) of swine-origin the functional implication of an amino acid (S186) present influenza virus H1N1 (S-OIV) reveals a number of atypical uniquely in the N terminus of the PA subunit of the pandemic characteristics including an early start codon and a highly H1N1 influenza virus and determined the effect of its mutation conserved, non-aromatic residue at position 313. Using an in in terms of polymerase activity as well as virus growth. Using vitro viral polymerase reconstitution assay, the polymerase chimeric constructs of PA derived from A/PR/8/34 (H1N1) (PR8) complex containing the NP of S-OIV (NPS-OIV) had substantially and the swine-origin influenza virus (S-OIV) H1N1 (complexed lower activity than those assays with NP derived from other with PB2 and PB1 of PR8), the chimeric PA protein containing influenza virus strains. Moreover, alteration of the early start the N terminus of S-OIV (1–213) with the remaining region codon or introduction of an aromatic residue at position 313 from PR8 showed significantly reduced polymerase activity. (V313Y) did not increase but instead exacerbated the poor Recombinant viruses harboring the chimeric PA also grew polymerase activity. When NPS-OIV was allowed to compete poorly in MDCK cells and embryonated eggs. Likewise, the with that of a mouse-adapted influenza virus (A/PR/8/34) to chimeric PA in which the N terminus of PA of PR8 (1–213) form progeny virions, only progeny bearing NPS-OIV were was assembled with the remaining region of PA of S-OIV produced, despite the low polymerase activity associated with showed a similar phenotype when complexed with PB2 and NPS-OIV. Results indicated that NPS-OIV requires both the early PB1 of S-OIV. Interestingly, when S186 in the N terminus was start codon and the V313 residue for its optimal function. These altered to the residue common in most strains of influenza characteristics are required for strong compatibility between virus (G186), the chimeric as well as wild-type PA of S-OIV the S-OIV polymerase subunits and its indigenous NP over showed severely impaired polymerase activity when assayed that of other strains, which might explain why productive with PB2 and PB1 of S-OIV. Collectively, this finding suggests reassortment between S-OIV and seasonal influenza viruses that S186 at the N terminal region of PA of S-OIV is necessary has yet to occur in nature. The data collectively indicated that for the protein to function optimally. This study is the first unique characteristics of NPS-OIV, including the early start knowledge to identify the functional implication of PAS186. codon and the V313 residue in NPS-OIV, are essential for the Results demonstrated that the strain-specific amino acid S186 compatibility between NPS-OIV and the polymerase complex at the N terminal region of the PA polymerase of S-OIV plays a of S-OIV, which could potentially contribute to restriction of critical role in determining the compatibility with the PB2 and reassortment between S-OIV and the influenza virus used in PB1 subunits of S-OIV. Given the potential for PAS186 to be the this study, and possibly other human influenza viruses as well. restriction factor for reassortment with polymerase complexes This work by the Virology and Cell Technology Laboratory was of other influenza viruses, the chance of S-OIV polymerase published in Archive of Virology in February 2011.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 29
Highlights on Medical Biotechnology
Novel immunomagnetic particles for Novel test tube method of screening for separation of CD4+ lymphocytes hemoglobin E The research team at the Biomedical Technology Research Hemoglobin (Hb) E is a b-structural variant common worldwide. Center, a joint research unit between BIOTEC and Chiang Mai This Hb disorder can form a compound heterozygous state with University, in collaboration with the National Nanotechnology the b-thalassemia gene, leading to life-threatening hereditary Center (NANOTEC), has developed novel immunomagnetic hemolytic anemia, HbE/b-thalassemia. Screening of HbE has particles for separation of CD4+ lymphocytes. The magnetic proven to be a challenging practice in prevention and control nanoparticles were successfully prepared by co-precipitation of the HbE/b-thalassemia. The research team at the Biomedical from ferrous and ferric iron solutions and subsequently Technology Research Center developed a novel test tube encapsulated with poly (glycidyl methacrylate) (PGMA) by method for HbE screening using diethyl aminoethyl (DEAE)- precipitation polymerization. Monoclonal antibody specific cellulose resin. With the developed system, HbE/A2 did not bind to CD4 molecules expressed on CD4+ lymphocytes was to the resin and remained dissolved in the supernatant, whereas covalently bonding between the epoxide groups on the other Hbs completely bound to the resin. The red color of the surface of particles and primary amine groups of antibody. supernatant observed in the test tube indicated the presence The generated immunomagnetic particles were specific of HbE. Colorless or markedly pale color of the supernatant and highly efficient in separation of CD4+ lymphocytes from indicated the absence of HbE. Accuracy and efficiency of the whole blood with over 95% purity. The results indicated that established method in detecting HbE was comparable with the these immunomagnetic particles can be useful for CD4+ standard cellulose acetate electrophoresis method. In addition, lymphocytes and other cells separation which provide strong this method is inexpensive and simple with no requirement for potential to be applied in various biomedical applications sophisticated equipment. The reagent could be stored at 4°C including treatment, diagnosis and monitoring of human for up to 5 months. Hemolysate samples aged up to 5 months diseases without the need for column separation technique. were still suitable for this test. The described novel test tube This work was reported in Talanta in March 2011. method could be an alternative method of mass population screening for HbE, particularly in small health care facilities. This work was published on-line in the International Journal of Laboratory Hematology August 2011.
Annual Report 2011 30
Highlights on 66.9% Bioresources Management
25.4% Microbial Cultures in BCC • 32,037 strains of filamentous fungi (66.9%) 7.3% • 12,164 strains of bacteria (25.4%) • 3,484 strains of yeasts (7.3%) 0.4% • 208 strains of algae (0.4%)
processing of bioassay data and field biology/ecology data to allow scientists to search for samples and activity relationships. More than 30,000 strains of microbes are now available in the MIMS database.
Data management system for bioresource utilization research A data management system for research in bioresource utilization has been developed in several projects by the research team at the Information Systems Laboratory of the Microbial culture collection Bioresources Technology Research Unit. Software has been Established in 1996, the BIOTEC Culture Collection has a developed and is currently used for supporting the laboratory’s primary task to collect and maintain microorganisms and inventory management of biological collections at BIOTEC their relevant data for BIOTEC’s in-house research. In addition using iCollect. It allows users to create any type of sample to the maintenance and distribution of microorganisms containers and storage devices, store any type of samples for in-house research, BCC provides culture deposit, safe and collections, graphically view the content of all levels of deposit and lyophilization services to public. The facility is storages, generate and print barcode, easily aliquot/derive/ also designated by the Thailand Department of Intellectual extract any number of samples and search any information in Property as a repository of patent-related microorganisms. the collection. Using the barcode assigned by iCollect, users Under international standard of preservation and database can easily find a sample’s position in the storage, and add a management (certified ISO9001:2008), BCC holds 47,893 sample to a selected storage position. The in and out movement strains of microorganisms as of October 2011, of which 67% of samples in the storage are recorded and tracked for audit are identified strains, while less than 33% are unidentified trail. In the graphical tree view of the samples, users can trace strains. Among these, 60 strains are patented-related and safe the aliquot/sample back to its original parent. deposited. In addition to microorganisms, BCC also preserves a total of 105 biological materials, comprising 38 vectors, 19 iCollect has also been installed at the Thailand Institute of hosts, and 48 recombinants. Scientific and Technology Research (TISTR) and National Science Museum (NSM) to manage and integrate biological The MIMS (Microbial Information Management System) databases including plants, microbes, and animals, covering has been established as an in-house database system for at least 100,000 pieces of biodiversity-related information managing the taxonomic, ecological, bibliographic and among agencies under the Ministry of Science and Technology graphic information of the microbial culture collection at (MOST). A Memorandum of Understanding (MOU) has been the Bioresources Technology Unit. The key features of MIMS signed for database collaboration among the three agencies include the storage of specimen-related information and the covering 3 years (2011- 2014).
National Center for Genetic Engineering and Biotechnology (BIOTEC) 31
Highlights on Microbial Research and Utilization
Multi-gene phylogeny of the and they proposed the erection of three new genera to Halosphaeriaceae accommodate C. tubulifera (Toriella), R. crispa (Kochiella) and R. galerita (Tubakiella). The molecular data indicated that different The halosphaeriaceous fungi constitute the largest group phylogenies based on DNA sequences support a hypothesis of marine Ascomycota found predominantly in marine that ascospore appendage developments have evolved and environments, with a few transitional species found in been lost several times. This work was published in Fungal freshwater and brackish water habitats. It has been one of Diversity in November 2010. the most intensively studied groups of marine fungi, with 126 species in 53 genera. The classification of the halosphaeriaceous New species of Ophiocordycepts fungi is contentious, with one school maintaining that they should be regarded as a family in the Microascales, while others halabalaenses in Hala-Bala Wildlife continue to retain the order Halosphaeriales. To refine the Sanctuary phylogenetic inter-relationships among the halosphaeriaceous In an ongoing biodiversity study of fungal pathogens on ants fungi, a joint research team comprising the Mycology Laboratory in several national parks in Thailand, previous research has of the Bioresources Technology Unit and the Institute of Marine shown that there were 7 species of fungal pathogens on ants. Biology of the National Taiwan Ocean University sequenced One of them is Ophiocordyceps unilateralis, which is a common and analysed 36 taxa based on three loci [nuclear small and ant pathogen found in Thailand. Surveys in Hala-Bala Wildlife large subunit (SSU, LSU), the second largest RNA polymerase Sanctuary located in the South of Thailand, by a research team II subunit (RPB2)]. The halosphaeriaceous fungi constitute at the Mycology Laboratory of the Bioresources Technology a monophyletic group and share a common ancestor with Unit has revealed that an ant fungal pathogen found on the Microascaceae. However, they share few morphological collected specimens is specific to a giant forest ant called characters. Based on morphological data the halosphaeriaceous Camponotus gigas. Characteristics of the ant-specific fungus taxa might be considered as a group warranting ordinal are morphologically similar to O. unilateralis by producing both status (Halosphaeriales). Sequence data showed clearly that a lateral pad on one side of the stroma and whole ascospores. the genera Remispora and Ceriosporopsis are polyphyletic Phylogenetic analyses of the partial elongation factor tef1-α
Annual Report 2011 32
gene sequences and 16S-23S rRNA gene internal transcribed spacer (ITS) sequences, (ii) the calculated 16S rRNA gene sequence and 16S-23S rRNA gene ITS sequence similarities, and (iii) DNA G+C contents showed that the strains clearly represent a novel species, for which the name Gluconobacter nephelii sp. nov. is proposed. This work was published in the International Journal of Systematic and Evolutionary Microbiology in September 2011. and the internal transcribed spacer regions ITS1-5.8S-ITS2 rDNA » Neokomagataea thailandica sp. nov. and have indicated that the ant-specific fungus is a distinct species Neokomagataea tanensis sp. nov. The research from O. unilateralis. The morphological characters of the fungus team has isolated and studied the new species of differ fromO. unilateralis in the possession of bigger perithecia acetic acid bacteria. The AH11T and AH13Tstrains and ascospores. Moreover, molecular analyses have shown were isolated from flowers of lantana and candle that this fungus is sufficiently different fromO. unilateralis. As bush respectively collected in Thailand. The a result, the ant-specific fungus discovered by the research analysis of (i) phylogenetic trees based on 16S team deserves to be a new species named Ophiocordyceps rRNA gene sequences, (ii) the calculated pair-wise halabalaensis. This work was published in Fungal Biology in 16S rRNA gene sequence similarities, and (iii) G+C March 2011. content revealed that Neokomagataea gen. nov. was proposed with the two novel species, which New species of acetic acid bacteria in a class are Neokomagataea thailandica sp. nov. for isolate of α-Proteobacteria AH11Tand Neokomagataea tanensis sp. nov. for isolate AH13T. This work was published in the Bioscience, Acetic acid bacteria are mostly studied in sugary acid or alcoholic Biotechnology, and Biochemistry in March 2011. sources, for example: beer, wine or fermented vinegar, but many new genera and species of acetic acid bacteria have New actinomycetes species in the genus of been isolated and proposed from other various sources in the past decade. Researchers from BIOTEC Culture Collection Herbidospora cretacea Laboratory in collaboration with Chulalongkorn University Over the past sixty years, a significant amount of effort has been and NITE Biological Resource Center in Japan discovered new focused on the isolation and identification of novel bioactive species of acetic acid bacteria in a class of α-Proteobacteria. metabolites from actinomycetes. It has been known that These novel species will be additional biological resources actinomycetes are prolific sources of bioactive compounds with for further investigation and utilization. Examples of new diverse chemical structures, which are shown to possess distinct species include: biological activities. In addition, actinomycete strains could be an excellent bioactive metabolite producer. Researchers » Gluconobacter nephelii sp. nov. The research team have isolated and studied an actinomycete strain DMKUA has isolated and studied three acetic acid bacterial 205T, which is isolated from a soil sample collected from the strains in the class of α-Proteobacteria, RBY-1(T), Sakaerat Biosphere Reserve in Nakhon Ratchasima Province. PHD-1 and PHD-2, from rambutan and lychee in On the basis of morphological, chemotaxonomic, phenotypic, Thailand. Their phenotypic and chemotaxonomic genotypic and phylogenetic data, strain DMKUA 205T could characteristics were investigated. The results revealed be clearly distinguished from the type strains of H. yilanensis that the strains were acetic acid bacteria. The and H. daliensis. Therefore, strain DMKUA 205T represents a analysis of (i) phylogenetic trees based on 16S rRNA novel species, for which the name Herbidospora sakaeratensis
National Center for Genetic Engineering and Biotechnology (BIOTEC) 33
sp. nov. is proposed. In addition, the DNA–DNA hybridization conoideocrellide A, its linear derivatives, results from this study revealed that Streptosporangium conoideocrellides B−D, three new hopane claviforme is a later synonym of Herbidospora cretacea. This triterpenoids and two new bioxanthracenes, and work was a collaboration of various partners, including BCC, a new isocoumarin glycoside were identified and Kasetsart University, NITE Biological Resource Center (NBRC) reported as new compounds. Biological activities and Shizuoka University, Japan. The report was published of the new compounds were evaluated. The result in the International Journal of Systematic and Evolutionary showed Triterpine 6 exhibited antiviral HSV-1 activity, Microbiology in April 2011. anti-oral epidermoid carcinoma activity (KB), anti- breast cancer activity (MCF-7) and anti-small cell New bioactive compound discovery lung cancer (NCI-H187). This work was published in the Journal of Natural Products in April 2011. Over 40 new and 33 known compounds from various microorganisms such as insect fungi, mushroom fungus including actinomycetes have been isolated and evaluated Functional characterization of truncated for their biological activities. These new compounds will be fragments of B. sphaericus binary toxin BinB new sources for further novel drug development. Examples Bacillus sphaericus produces a mosquitocidal protein, called of novel compounds are: binary toxin. This toxin is composed of two subunits, BinA and BinB which contain 42 kDa and 51 kDa, respectively. » Sterostreins A-E, New Terpenoids from cultures Both components are required for maximum toxicity against of the Basidiomycete Stereum ostrea BCC 22955 mosquito larvae. BinB has been proposed to provide specificity Sterostreins A-E, five novel terpenoids, were isolated by binding to the epithelial gut cell membrane, while BinA from cultures of the mushroom fungus Stereum may be responsible for toxicity. To identify regions in BinB ostrea BCC 22955. Sterostrein A exhibited antimalarial which are responsible for receptor binding and interaction activity and cytotoxicity. This work was published to BinA, researchers have examined the binding of six BinB in the Journal of Organic Chemistry in August 2011. shorter constructs derived from both the N-terminal and the » Antimalarial and antituberculosis substances C-terminal halves of the protein to the epithelium gut cell from Streptomyces sp. BCC26924 Three new membrane and the binding region between BinA and different compounds (carbazomycin dimmers) were isolated parts of the N-and C- halves of BinB. The result revealed that from actinomycetes and six known compounds fragments corresponding to both N- and C- halves of BinB are (cyclomarin C, carbazomycins A-D and pimprinine). able to bind the receptor and interact with BinA. The binding of Carbazomycins B and C, cyclomarin C showed cell surface receptor on epithelial gut cell membrane required antimalarial activity (against Plasmodium falciparum, several amino acids on BinB. In addition, the experiment K1 multi-drug resistant strain). In addition, cyclomarin indicated that the amino acid arrangement on BinB is critical C exhibited anti-TB activity with a minimum to the binding between BinA and BinB. However, both C inhibitory concentration value of 0.10 μg/mL, while and N-halves are required by the binary toxin to exhibit full carbazomycin D displayed MIC values in a range of larvicidal activity. The knowledge gained from this research 12.5-25.0 μg/mL. This work was published in the will lead to further study on the function and mechanism of Tetrahedron in September 2011. the binary toxin, BinA and BinB, for killing the mosquito larvae. » Bioactive compounds from the scale insect This work, a collaboration between the Microbial Engineering pathogenic fungus Conoideocrella tenuis BCC Laboratory of the Bioresources Technology Unit and Mahidol 18627 Ten new compounds were isolated from University, was published in the Journal of Invertebrate Pathology the insect fungus Conoideocrella tenuis BCC18627. in October 2010. The structure of new cyclohexadepsipeptide,
Annual Report 2011 34
Genes involved in NQ production from Phylogeny and metabolic potential of Ophiocordyceps unilateralis a primary tropical peat swamp forest Napthoquinones (NQs) is a secondary metabolite useful in microbial community the medical application and found in ant-pathogen fungi. BIOTEC research team conducted a study on the phylogeny One of them is Ophiocordyceps unilateris BCC1869 which can and metabolic potential of a primary tropical peat swamp forest produce six NQs derivatives. Also, the NQs have antibacterial, microbial community by metagenomic analysis. A primary anticancer, and antimalarial activities and red pigments with tropical peat swamp forest is a unique ecosystem characterized potential for use as natural colorants. The research team at the by long-term accumulation of plant biomass under high Bioresources Technology Unit conducted a study to identify humidity and acidic water-logged conditions and is regarded genes potentially involved in biosynthesis and secretion as an important terrestrial carbon sink in the biosphere. In of NQs in fungus O. unilateralis BCC1869. To identify genes this study, the microbial community in the surface peat layer differentially expressed between fungal cultures producing in Pru Toh Daeng, a primary tropical peat swamp forest, was NQs and cultures lacking NQs, the suppressive subtractive studied for its phylogenetic diversity and metabolic potential hybridization (SHH) method was performed. Results showed using direct shotgun pyrosequencing of environmental that there were sixty-six unique expressed sequence tags DNA, together with analysis of 16S rRNA gene library and key (ESTs) which include five transporter genes, two transcriptional metabolic genes. The community was dominated by aerobic regulator genes and several genes in secondary metabolism microbes together with a significant number of facultative and biodegradation. The transporter genes include an and anaerobic microbial taxa. Acidobacteria and diverse ATP-binding cassette transporter gene OuAtr1 and a major Proteobacteria (mainly Alphaproteobacteria) constituted facilitator super family transporter gene OuMfs1. Expression the major phylogenetic groups, with minor representation of selected ESTs was further validated using quantitative of archaea and eukaryotic microbes. Based on comparative reverse transcription PCR. Gene expression results indicate pyrosequencing dataset analysis, the microbial community that OuAtr1 and OuMfs1 were dramatically upregulated (136- showed high metabolic versatility of plant polysaccharide and 29-fold increase, respectively) during the NQ–producing decomposition. A variety of glycosyl hydrolases targeting stage compared with the NQ–nonproducing stage. This lignocellulosic and starch-based polysaccharides from diverse EST collection represents the first group of genes identified bacterial phyla were annotated, originating mostly from from this potential biocontrol agent and includes candidate Proteobacteria and Acidobacteria, together with Firmicutes, genes for production and secretion of the red NQs. Roles of Bacteroidetes, Chlamydiae/Verrucomicrobia and Actinobacteria, these genes could be further determined using a functional suggesting the key role of these microbes in plant biomass analysis which is either a gene knocking out system or RNA degradation. Pyrosequencing dataset annotation and direct silencing. This work was published in the Canadian Journal of mcrA gene analysis indicated the presence of methanogenic Microbiology in August 2011. archaea clustering in the order Methanomicrobiales, suggesting the potential on partial carbon flux from biomass degradation
National Center for Genetic Engineering and Biotechnology (BIOTEC) 35
through methanogenesis. The insights on the peat swamp microbial assemblage thus provide a valuable approach for further study on biogeochemical processes in this unique ecosystem. This study was jointly conducted by three BIOTEC research units, namely Genome Institute, Bioresources Technology Unit and Food Biotechnology Research Unit. This work was published in Microbial Ecology in April 2011 and highlighted on A-IMBN Research.
Xylanase production from Aspergillus niger Comparative analysis of Spirulina platensis for pulp bleaching subcellular proteome Biobleaching of pulp using xylanases is recognized as an Spirulina cells encounter temperature fluctuations, associated economically feasible biotechnology-based process in the pulp with outdoor mass cultivation, that have an effect on and paper industries. Pre-bleaching with xylanases reduces the biomass yield and the biochemical content of the cells. Some need for toxin bleaching chemicals and is thus environmentally components with pharmaceutical benefits, such as unsaturated and economically advantageous. Enzyme-aided bleaching is fatty acids in membrane lipids, have been shown to play vital the first full-scale biotechnological application in the pulp and roles in the response to temperature change. In the study paper industry. In order to enhance chemical delignification conducted by BIOTEC in collaboration with King Mongkut’s efficiency, xylanases are used to modify the structure of University of Technology Thonburi, scientists focused on xylan and glucomannan in pulp fibers. BIOTEC researchers, in comparative proteome analyses of low- and high-temperature collaboration with SCG Paper PLC, have optimized xylanase stresses and potential protein-protein interaction networks, production from the fungus Aspergillus niger BCC14405 for constructed by using a bioinformatics approach, in response pulp bleaching. A crude endo-xylanase produced by A. niger to both stress conditions. The data revealed two important BCC14405 was investigated for its potential in pre-bleaching points: first, the results indicate that low-temperature stress is of chemical pulp from eucalyptus. The results obtained from tightly linked with oxidative stress as well as photosynthesis; optimal fermentation conditions showed xylanase production however, no specific mechanism is revealed in the case of was 89.5IU/mL with minor cellulose activity. Also, the crude the high-temperature stress response. Second, temperature enzyme was a composite of other hemicellulolytic enzymes stress was revealed to be linked with nitrogen and ammonia attacking arabinoxylan and mannan. Pretreatment of the assimilation. Moreover, the data also highlighted the cross-talk pulp at a xylanase dosage of 10IU/g increased the brightness of signaling pathways. Some of the detected signaling proteins, ceiling. Moreover, xylanase treatment led to reduction in e.g., Hik14, Hik26 and Hik28, have potential interactions with chlorine charge of at least 20%, with an acceptable brightness differentially expressed proteins identified in both temperature level. This research has shown the crude enzyme from stress conditions. Some differentially expressed proteins found A. niger BCC14405 has potential use in efficient pretreatment in the Spirulina protein-protein interaction network were also of eucalyptus pulp prior to industrial bleaching. The results examined for their physical interactions by a yeast two hybrid obtained for eucalyptus pulp in this study are of industrial system (Y2H). The Y2H results suggest that the potential PPI significance and provide a promising alternative to on-site network gives quite reliable potential interactions for Spirulina. enzyme production for the development of a sustainably This study was published in Proteome Science in July 2011 and greener pulping industry. This work was published in Bioscience, highlighted on A-IMBN Research. Biotechnology, and Biochemistry in June 2011.
Annual Report 2011 36 Highlights on Biorefinery
Enhancing UASB treatment of palm oil mill effluent using chitosan The effects of chitosan addition on treatment of palm oil mill effluent in an upflow anaerobic sludge bed (UASB) reactor were Highlight on Ecology investigated in a collaborative project between the Excellent Center of Waste Utilization and Management, a joint laboratory Non-redundancy in the dispersal network of between BIOTEC and King Mongkut’s University of Technology a generalist tropical forest tree Thonburi (KMUTT) and King Mongkut’s University of Technology Plant species with generalized dispersal mutualisms are North Bangkok. It was found that chitosan could enhance the considered to be robust to local frugivore extinctions because sludge granulation process and UASB performance, especially of redundancy between dispersal agents. However, real in terms of effluent volatile suspended solids (VSS) and biogas redundancy can only occur if frugivores have similar foraging production. Chitosan also helped to retain the methanogens in and ranging patterns and if fruit is a limiting resource. Joint the genus Methanosaeta, thus resulting in higher populations research between the Ecology Laboratory and a researcher from of acetotrophic methanogens. This work was published in India evaluated the quantitative and qualitative contributions Bioresource Technology in January 2011. of seed dispersers for an endochorus mast-fruiting species, Prunus javanica (Rosaceae) in Khao Yai National Park, Thailand. Biocatalytic esterification of palm oil fatty To evaluate the potential redundancy of dispersers, data acids for biodiesel production were collected from tree watches, seed/fruit traps and seed In biodiesel production, the conversion of feedstock containing transects under and away from fruiting trees, feeding and seed high free fatty acid contents to alkyl esters is limited by the deposition by gibbons (Hylobates lar), and evaluations of seed currently used alkali-catalyzed biodiesel synthesis process. In a and first-year seedling survival. The research team identified joint study conducted by the Enzyme Technology Laboratory of three clusters of dispersers within the network. Firstly, most the Bioresources Technology Unit and KMUTT, esterification of (>80%) frugivore species observed were small birds and squirrels palm fatty acids to ethyl esters was studied using heterogeneous that were not functional dispersers, dropping most seeds cross-linked protein coated microcrystalline (CL-PCMC) lipase. under or very near the tree crown, where seedling survival Highly active CL-PCMCs could be prepared from Thermomyces was ultimately nil. Secondly, monkeys (Macaca leonina) were lanuginosus lipase with glycine as the core matrix in acetone. low-quality, short-range dispersers, but they dispersed large This CL-PCMC-lipase was found to be more catalytically numbers of seeds and were responsible for 67% of surviving efficient than protein coated microcrystalline (PCMC) lipase, first-year seedlings. Lastly, gibbons and Oriental Pied Hornbills Novozyme®435 and Lipolase 100T, for both free fatty acids and (Anthracoceros albirostris) handled few fruits, but they provided palm fatty acid distillate. The glycine-based microcrystalline the highest quality service by carrying most seeds away from lipase is thus a promising alternative economical biocatalyst the canopy to medium and long distances, respectively. for biodiesel production from inexpensive feedstock with high Although there was overlap in the deposition patterns of the free fatty acid contents. This would lead to an improvement in functional dispersers, they displayed complementary, rather the process economics of the biodiesel industry. This work was than redundant, roles in seed dispersal. The results underscore filed for Thai patent and published in theJournal of Molecular the need for research on broader spatial and temporal scales, Catalysis B: Enzymatic in August 2011. which combines studies of dispersal and plant recruitment, to better understand mechanisms that maintain network stability. This work was published in Ecology in July 2011.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 37 Highlights on Bioinformatics and Information Systems
LinkinPath: a web-based interactive exploration tool for pathway analysis Pathways are functional units resulting from the interplay of interacting genes, RNAs, proteins and small molecules. Mapping genes or proteins into the context of pathways can help gain more insights into their functions and interactions in an organism. The research team at the Information Systems Laboratory has developed LinkinPath, a pathway mapping and analysis tool that enables users to explore and visualize the list of gene/protein sequences through various Flash- driven interactive web interfaces including KEGG pathway maps, functional composition maps (TreeMaps), molecular interaction/reaction networks and pathway-to-pathway networks. Users can submit single or multiple datasets of gene/protein sequences to LinkinPath to (i) determine the co-occurrence and co-absence of genes/proteins on animated KEGG pathway maps, (ii) compare functional compositions within and among the datasets using TreeMaps, (iii) analyze Consortium, constructed a SNP database (PanSNPdb), which the statistically enriched pathways across the datasets, (iv) contains data and various new analyses. PanSNPdb is a research build the pathway-to-pathway networks for each dataset, resource in the analysis of the population structure of Asian (v) explore potential interaction/reaction paths between peoples, including linkage disequilibrium patterns, haplotype pathways, and (vi) identify common pathway-to-pathway distributions and copy number variations. Furthermore, networks across the datasets. Therefore, using LinkinPath to PanSNPdb provides an interactive comparison with other overlay new sequence data from any system of interest onto SNP and CNV databases, including HapMap3, JSNP, dbSNP established pathways and networks will aid understanding of and DGV and thus provides a comprehensive resource of the regulation of cellular signaling and metabolism. LinkinPath human genetic diversity. The information is accessible via a is freely available to all interested users at http://www.biotec. widely accepted graphical interface used in many genetic or.th/isl/linkinpath/. This work was published in Bioinformatics variation databases. Unrestricted access to PanSNPdb and in July 2011 and was highlighted on A-IMBN Research. any associated files are available at: http://www4a.biotec. or.th/PASNP. This work was published in PLoS ONE in June PanSNPdb: The Pan-Asian SNP genotyping 2011 and was highlighted on A-IMBN Research. database Study of population datasets by combining The HUGO Pan-Asian SNP consortium conducted the largest survey to date of human genetic diversity among Asians by ipPCA and STRUCTURE sampling 1,719 unrelated individuals among 71 populations A new population stratification testing heuristic, called EigenDev, from China, India, Indonesia, Japan, Malaysia, the Philippines, was proposed by the a team composed of researchers from Singapore, South Korea, Taiwan and Thailand. This work the Genome Institute, King Mongkut’s Institute of Technology continued from the research titled Mapping Human Genetic Ladkrabang and the National Electronics and Computer Diversity in Asia published in Science in 2009. The research Technology Center (NECTEC). This heuristic markedly improves team at the Genome Institute and Medical Molecular Biology the performance of the previously published population Research Unit, in collaboration with the HUGO Pan Asian SNP classification algorithm by the team at Genome Institute, called
Annual Report 2011 38
iterative pruning principal component analysis (ipPCA). When humans. Both peptides were able to inhibit ADE, showing that tested on simulated data, this proposed population structure entry inhibitors are possible candidates for the development testing heuristic is robust to sample size. In contrast, the of specific treatment for severe DENV infection. This work was popular Tracy-Widom (TW) statistical test was demonstrated published in Antiviral Research in January 2011. in this work to be susceptible to type I error, especially for large population samples. EigenDev is thus better-suited for analysis Hypomethylation of intragenic LINE-1 of large datasets containing many individuals, in which spurious represses transcription in cancer cells patterns are likely to exist and could be incorrectly interpreted through AGO2 as population stratification. This resulting subpopulation information was used to supervise STRUCTURE analysis to infer In human cancer, the methylation of long interspersed patterns of ancestry at an unprecedented level of resolution. nuclear element-1 (LINE-1 or L1) retrotransposons is reduced, In summary, the EigenDev heuristic is robust to sampling and also called hypomethylation. The role of this genome wide is thus superior for detecting structure in large datasets. The hypomethylation is poorly understood, albeit common. The application of EigenDev to the ipPCA algorithm improves research team at the Genome Institute in collaboration with the estimation of the number of subpopulations and the a team from Chulalongkorn University, studied the role of individual assignment accuracy, especially for very large and intragenic L1 hypomethylation that would influence their complex datasets. Furthermore, the structure resolved by this host gene expression in cancer. The results demonstrated approach complements parametric analysis, allowing a much that intragenic L1 sequences have been conserved across more comprehensive account of population structure. As a evolutionary time with respect to transcriptional activity result, the combination of EigenDev-ipPCA and STRUCTURE and CpG dinucleotide sites (CG sites) for mammalian DNA are complementary and can be used together to perform a methylation. The regulated mRNA levels of cells from two powerful population stratification analysis. The new version different experiments available from Gene Expression Omnibus of the ipPCA software with EigenDev incorporated can be (GEO) were statistically evaluated by chi-square. Both odds ratio downloaded from http://www4a.biotec.or.th/GI/tools/ippca, and p-value of down-regulated genes between demethylated and this work was published in BMC Bioinformatics in June 2011. normal bronchial epithelium and lung cancer were markedly significant, suggesting cancer genome wide hypomethylation Viral entry inhibitors block dengue down-regulating gene expression. Furthermore, comprehensive antibody-dependent enhancement in vitro analysis between L1 locations and gene expression showed that expression of genes containing L1s had a significantly Severe dengue virus (DENV) disease symptoms, including higher likelihood to be repressed in cancer and hypomethylated dengue hemorrhagic fever and dengue shock syndrome, have normal cells. In contrast, many mRNAs derived from genes been correlated with the presence of pre-existing antibodies containing L1s are elevated in Argonaute 2 (AGO2 or EIF2C2)- that enhance rather than neutralize infections in Fc receptor depleted cells. Hypomethylated L1s increase L1 mRNA levels. bearing cells. These antibodies can originate from previous AGO2 targets intronic L1 pre-mRNA complexes and represses infection with a different serotype of dengue, or from waning cancer genes. These findings represent one of the mechanisms antibody titers that occur in infants and young children as they of cancer genome wide hypomethylation altering gene are weaned from breast milk that contains protective dengue- expression. Hypomethylated intragenic L1s are a nuclear siRNA specific antibodies. Despite the apparent importance of this mediated cis-regulatory element that can repress genes. This antibody dependent enhancement (ADE) effect, there has epigenetic regulation of retrotransposons likely influences been no description of any specific inhibitors of this process. many aspects of genomic biology. This work was published The research team at the Genome Institute, in collaboration in PLoS ONE in March 2011. with a team at the University of Washington, investigated DENV entry inhibitors as potential candidates to block dengue ADE. Two different peptide entry inhibitors were tested for the ability to block antibody-mediated DENV-2 infection of
National Center for Genetic Engineering and Biotechnology (BIOTEC) 39 Highlights on Proteomics
Secreted Opisthorchis viverrini glutathione Phosphoproteomic analysis of apoptotic S-transferase regulates cell proliferation in hematopoietic stem cells from hemoglobin cholangiocarcioma E/β-thalassemia Opisthorchis viverrini is the strong risk factor of Hemoglobin E/b-thalassemia is particularly common in cholangiocarcinoma (CCA) in Thai populations where this Southeast Asia and has variable symptoms ranging from human liver fluke is endemic. Opisthorchis viverrini can develop mild to severe anemia. Previous investigations demonstrated mitogenic substances into the excretory/secretory product the remarkable symptoms of b-thalassemia in terms of the (ESP) that may play an important role in promoting the genesis acceleration of apoptotic cell death. Ineffective erythropoiesis of cholangiocarcinoma (CCA). Researchers conducted a study has been studied in human hematopoietic stem cells, however on secreted opisthorchis viverrini glutathione s-transferase the distinct apoptotic mechanism was unclear. Researchers regulation of cell proliferation through AKT andERK pathways from the Genome Institute, Department of Biochemistry, in cholangiocarcioma. In this study, glutathione S-transferase Department of Pediatrics, Faculty of Medicine Ramathobodi (GST) is identified as being secreted into Ov-ESP and acting Hospital, Mahidol University and Faculty of Science and as one of the parasiticmitogens. Its proliferative effect and Technology, Suan Dusit Rajabhat University conducted possible mechanism were explored and its association with research on phosphoproteomic analysis of apoptotic tumor development was postulated. Ov-ESP was concentrated hematopoietic stem cells from hemoglobin E/beta-thalassemia. and purified by gel filtration chromatography. SDS-PAGE, 2-DE The phosphoproteome of bone marrow HSCs/CD34+ cells and LC-MS/MS identified GST predominantly expressed in the from HbE/b-thalassemic patients was analyzed using IMAC proliferative ESP fraction. The recombinant OvGST (rOvGST) was phosphoprotein isolation followed by LC-MS/MS detection. produced by wheat germ cell-free expression and confirmed Decyder MS software was used to quantitate differentially by an MTS assay to have a proliferative function on NIH-3T3 expressed proteins in 3 patients and 2 normal donors. The murine fibroblasts and MMNK1 non-tumorigenic human differentially expressed proteins from HSCs/CD34+ cells bile duct epithelial cells in a dose dependent manner with were compared with HbE/b-thalassemia and normal HSCs. different optimal doses. The cell surface binding of rOvGST A significant change in abundance of 229 phosphoproteins was confirmed in vitro and the activation of both pAKT and was demonstrated. Importantly, the analysis of the candidate pERK was revealed as the mechanism of OvGST-mediated cell proteins revealed a high abundance of proteins that are proliferation. With support from the observation of secreted commonly found in apoptotic cells including cytochrome C, OvGST on the biliary cells surrounding the parasites, it is caspase 6 and apoptosis inducing factors. Moreover, in the HSCs suggested that OvGST can promote cell proliferation that patients a significant increase was observed in a specific type consequently may accelerate the genesis of CCA. This is a of phosphoserine/threonine binding protein, which is known joint project between scientists from the Genome Institute, to act as an important signal mediator for the regulation of cell Department of Biochemistry of Khon Kaen University and survival and apoptosis in HbE/b-thalassemia. This study used a the Liver Fluke and Cholangiocarcinoma Research Center novel method to investigate proteins that influence a particular of Khon Kaen University. The work was published on-line in pathway in a given disease or physiological condition. Also, Parasitology International in July 2011. this report provides a comprehensive phosphoproteome, an important resource for the study of ineffective erythropoiesis and developing therapies for HbE/b-thalassemia, and it was published in Journal of Translational Medicine in June 2011.
Annual Report 2011 40 Highlights on Biosafety
Biosafety guidelines BIOTEC, as Secretariat of the Technical Biosafety Committee (TBC), launched new Biosafety Guidelines for Contained Use of Genetically Modified Microorganisms at Pilot and Industrial Scales in January 2011. The Guidelines provide the basic biosafety information, clear perspective on key issues, regulations and guidelines for using genetically modified microorganisms at pilot and industrial scales. This includes topics such as containment measures, biosafety levels, risk assessment and management, basis for the classification of biohazardous agents by risk group, certified host-vector systems Mapping airborne pollen of papaya and its and good laboratory and industrial large scale practices. A revision of the Biosafety Guidelines for Work Related to Modern distribution Biotechnology or Genetic Engineering was also published in Papaya is an economically important crop in Thailand for both 2011. The revised edition provides relevant basic biosafety domestic consumption and export. However, it is extremely information, general principles and framework for work susceptible to disease caused by the papaya ring spot virus related to modern biotechnology or genetic engineering. (PRSV). Although transgenic papaya has been developed Topics covered in these Guidelines include types of research by BIOTEC researchers at the Plant Research Laboratory, or experimentation involving genetically modified organism commercial cultivation of transgenic plants in Thailand is still (GMOs), routine precautions when working with biohazards, illegal. One concern is cross-pollination to conventional varieties. disinfectants, disposal of biohazards, laboratory biosafety Researchers have investigated windborne-pollen dispersion and biological levels as well as risk assessment for lab and of papaya (Carica papaya L.) using geographic information field experiments of genetically-modified microorganisms systems (GIS) and remotely sensed data. Pollen traps, filed for and plants. Thai petty patent in 2010, were placed around a papaya plot in eight geographic directions, with radiuses varying from 5 Status and trend of the use of GM to 900 meters away from the plot. Pollen counts on different microorganisms in Thai industry dates were put into a GIS database. Comparative analyses of pollen dispersal, wind direction and speed were made using BIOTEC was commissioned by the Department of Industrial data collected from a micro-climatic station set up at a papaya Works to conduct a study on the use of genetically-modified plot. The result showed that the furthest distance from the (GM) microorganisms in Thai industry. The study, completed plot that pollen was found was at 0.9 kilometers, a distance at in 2011, reviewed the current status and trends in the use of which only 1 pollen grain was found. Moreover, the number of GM microorganisms in various types of industry. The report pollen grains carried by wind decreased as distance increased. will also provide recommendations on the management and The direction of dispersal was not in accordance with wind regulation framework to ensure the safety to the environment direction data. Most pollen grains were found in agricultural and human health. The study will be useful to further develop areas and bare land. The total number of pollen grains found biosafety guidelines for each type of industry. in exposed areas was considerably higher than the total found in areas sheltered by dense tree lines. This study was jointly conducted by BIOTEC (Geoinformatics Laboratory and Plant Research Laboratory), Kasetsart University and King Mongkut’s University of Technology Thonburi. The work was published on-line in Aerobiologia in February 2011.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 41 Highlights on Policy Research and Development
Success indicators for technology transfer projects Success indicators for transferring technology projects were The result of this study was adopted by NSTDA to develop a developed to be used as a tool for more effective management strategy plan for its program on research, development and of NSTDA rural development programs. technology transfer for rural development. Moreover, the Research and Development Institute of Khon Kaen University, The indicators chosen by the communities show that the a project advisor, has planned to initiate a social process activities supported by NSTDA have assisted in community promoting project in Na Haew area, Loei Province in the future. development in 4 aspects; 1) Economic aspect – evaluated by an increase of Identifying NSTDA research and income, an establishment of new community development direction in Water Footprint enterprises/groups 2) Social aspect – assessed by related groups in Recently, Water Footprint (WF) and Virtual Water Trade (VWT) communities, strength of the group in bargaining have become new tools for quantitative assessment and impact power and accessing the support from other evaluation of production and consumption on freshwater. As organizations, job creation for informal laborers the consumer pays more attention to environmentally friendly e.g. housewives, students, elderly people. production, WF is used to show consumers and investors that 3) Environmental aspect – evaluated by positive and the product is responsible for maintaining water protection. negative environmental impacts from promoted activities The project aimed to identify the effect of this new footprint 4) Human resource development aspect – assessed on Thai export products and R&D topics where NSTDA by an increasing number of experts and leaders in should place priority. The result of the study could help the communities. Sustainable Environment Program to develop an appropriate The communities in each area also indicated that the strategy to promote and support research on WF for academia supported activities result in an increase of community and manufacturers. groups and community income. Furthermore, the supported activities in Hala Bala area, Narathiwat Province, showed The study indicates that the top five agricultural Thai exports considerable success in environmental management and may receive beneficial impact from the WF scheme. Other human development. Notable is the increase in local experts industrial products that consume large amount of water in and leaders, but also the youth who intend to come back production, e.g. textile, pulp and paper, automobile, are also for working in local areas after their graduation. With the included. In order to maintain Thailand’s competitiveness as determination of leaders and communities as well as support one of the world leaders in agricultural export in the coming from other organizations are also key drivers for the achievement WF scheme, research on WF assessment and improved water- of the activities. efficiency in production is critically needed. Moreover, it is suggested that NSTDA should play a key role as a focal point and Overall, all activities funded by NSTDA contribute to a satisfactory drive the related activities such as human resource development success in local development. However, those activities on WF assessment and WF consortium establishment. These cannot meet community needs in some aspects, especially will help promote multidisciplinary R&D and cooperation on in improving their main livelihoods, health and environment. joint activities under the same goal to achieve the national WF. Social process is essentially required to strengthen group and community power in order to enhance the sustainability of local development.
Annual Report 2011 42
COMMERCIALIZATION AND PRIVATE SECTOR PARTNERSHIP
National Center for Genetic Engineering and Biotechnology (BIOTEC) 43
IOTEC places strong emphasis on promoting the industrial Bapplication of biotechnology in both Thai and foreign companies. Technology/product licensing as well as collaborative research with the private sector are among mechanisms to promote bio-business in Thailand. Recognizing that the majority of companies in Thailand are of small and medium size and do not have in-house R&D capability, BIOTEC also provides R&D services in the form of contract research. In 2011 fiscal year, BIOTEC worked with 27 companies on 35 projects in the form of collaborative research and commissioned research, 9 of which were initiated in 2011. BIOTEC provided consultancy services to 18 companies on 18 different projects, 4 of which were new in 2011. Examples of collaborators and clients are Pfizer (Thailand) Limited, Arysta LifeScience Corporation (Japan), Excel Orchid Co., Ltd., Mitr Lao Sugar Co., Ltd., Siam Bioscience Co., Ltd. and Sitthinan Co., Ltd. Projects vary from identifying enzymes for alternative energy production, biocontrol production, utilization of wastewater from mungbean starch production, development of culture media to improve biomass and lactic acid production in fermentation process, developing a closed plant production system (CPPS) to providing advice on setting up a tissue culture laboratory for sugarcane.
Annual Report 2011 44
Thailand Biotech Guide 2010/2011
hailand Biotech Guide is a publication by Green World The 2010/2011 edition aims to highlight the importance of R&D TPublication Co., Ltd. and Marshall Cavendish Business in contributing to the continued development of Thailand’s Information Private Limited, with support from BIOTEC, bioindustry. The editorial section features companies and NSTDA and Thailand Board of Investment (BOI). Published in organizations such as Betagro Group, i+MED Laboratories Co., English, the Guide aims to serve as a biotech industry source Ltd., Microinnovate Co., Ltd. and PTT Chemical Plc. that have for foreign investors and promote foreign investments, with succeeded in developing innovative products and services, editorial articles on up-to-date biotechnology research and and bringing these innovations to the market in Thailand investment, a directory of biotech companies, as well as a and beyond. These examples will give readers more insight listing of suppliers for equipment and products supporting into the factors that contribute to successful R&D capability bio-industry. Thailand Biotech Guide has been published development and commercialization in Thailand. annually since 2006.
Licensing Agreements
ive licensing agreements have been accomplished with o BioNet-Asia Co., Ltd. The agreement enables BioNet-Asia Fthe following partners: Co., Ltd. to further develop four serotypes of chimeric live- attenuated vaccine into a commercial product. The University of North Texas Health Science Center o Asia Star Animal Health Co., Ltd. (ASAH) ASAH was at Forth Worth, USA The University obtained no-charge o licensed the technology to increase activity of non-starch permission to use iCollect, a software package to support the polysaccharide (NSP) degrading enzymes. ASAH previously laboratory’s inventory management of biological collections, obtained a license from BIOTEC for the production technology of for research purposes. pentosanase from Aspergillus sp. BCC7178 for feed supplement. o Aquatica Co., Ltd. Under the agreement, Aquatica has The new technology will improve the company’s productivity the rights to market a tubular denitrification reactor (TDNR) and improve stability of the feed product. technology for nitrate removal. With TDNR, water can be Hi-Grimm Environmental and Research Co., Ltd. Hi- recirculated within the system hence water exchange can o Grimm Environmental and Research was granted the rights to be prolonged. Aquatica is an aquarium retailer and provides IP management and commercialization of a research project to aquarium setting services. search for oil-degrading bacteria to develop into commercial bioremediation products.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 45
Chimeric Live- attenuated Dengue Vaccines
he four serotypes of dengue vaccine candidate were On 21 February 2011, NSTDA, Chiang Mai University, Mahidol Tjointly developed by scientists from Chiang Mai University, University and BioNet-Asia Co., Ltd. announced a licensing BIOTEC Medical Biotechnology Research Unit and Mahidol agreement for a chimeric dengue vaccine. The agreement University. The research team has successfully engineered enables BioNet-Asia Co., Ltd. to further develop and produce the chimeric live-attenuated vaccine for four serotypes of the vaccine for testing in pre-clinical and clinical stages of dengue viruses. A number of modifications of genetic material vaccine development into a commercial product. in different regions of the genome of dengue virus serotype 2 were found to result in the attenuation of virus virulence. BioNet-Asia Co., Ltd. is a biotech company based in Thailand with This discovery led to the construction and testing of the core business in the development and marketing of vaccines in chimeric viruses which are comprised of the prM-E coding the emerging countries. It is expected that this collaboration region from recent dengue clinical isolates on the genetic will lead to a commercial dengue vaccine available to Thais background of attenuated virus. Based on promising results and eventually expanded to the ASEAN market and beyond. from the neurovirulence and immunogenicity testing in mice, the chimeric viruses are considered to have good potential to be developed further as vaccine candidates for the safety and efficacy studies in a non-human primate model.
Annual Report 2011 46
New Products Available at BIOTEC SBBU in 2011
n 2011, BIOTEC’s Shrimp Biotechnology Business fruit blotch disease in curcubit, genetic tests for farm animal IUnit (SBBU) acquired licences for three new products and pests and a continuous microalgal culture system. invented by BIOTEC researchers. The three products are an These products and test services are available for interested immunochromatographic strip test for detection of bacterial customers through SBBU.
Continuous Microalgal Culture System IC Strip Test for Detection of Bacterial Fruit The photobioreactor is designed for continuous production of Blotch Disease in Cucurbit various species of microalgae such as Chaetoceros, Thalassiosira, The patented immunochromatographic strip test based on Chlorella and Spirulina. With the production capacity of mouse monoclonal antibodies against Acidovorax avenae 6 6-7L, at approximately 1x10 cells/mL concentration for subsp. citrulli – the bacteria which causes fruit blotch disease Chaetoceros which can be used as stock culture for further in cucurbit (cantaloupe, cucumber, watermelon, gourd, squash mass cultivation, the unit can operate continuously for 30 days and pumpkin) – has been developed to provide seed exporters before maintenance. It is equipped with the inline device for and their growers a tool to cost-effectively, conveniently, protozoa and microalgae decontamination, thus eliminating accurately and rapidly (in 5 to 10 minutes) detect the disease. the need of autoclave sterilization. This Continuous Microalgal The test enables epidemiological studies, disease management Culture System is especially suitable for shrimp hatcheries that and quality control for export of disease-free seed. require daily consumption of Chaetoceros as live feed for shrimp.
Genetic Tests for Farm Animal and Pests Available tests include: porcine growth rate, porcine meat tenderness and meat quality, porcine production performance and litter size, bovine marbling fat and meat tenderness, porcine resistance to E. coli and gene defects associated with canine drug allergy.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 47
BIOTEC @ NSTDA Investors’ Day
STDA Investors’ Day was organized on 22 September N2011 to showcase technologies and products to potential investors, with activities including Investment Pitching Session and One-on-One Matching. ENZbleach, an alkaline- tolerant enzyme for pulp bleaching process, was one the five technologies highlighted at this year’s event. In addition, four technologies were showcased, namely Lysis Buffer, Tubular Denitrification Reactor (TDNR), Continuous Microalgal Culture System, EzeeGene® Speedy Color Kit for shrimp disease detection (currently available for WSSV and TSV).
ENZbleach, an alkaline-tolerant enzyme for pulp bleaching process In a conventional chemical bleaching process, a large volume of chemicals is used, releasing pollution into the environment and causing numerous harmful disturbances to the ecosystem. The new biobleaching process using enzymes leads to a significant reduction of chlorine and chlorine-based compounds. The xylanase attacks hemicellulose and alters the interface between the cellulose and lignin, thereby facilitating the chemical extraction of lignin from pulp. For biobleaching applications, the candidate xylanase should be alkalophilic due to the high pH of the pulp. However, treatment of pulp with commercial xylanases at present needs a pH adjustment of the incoming pulp from 9.0-10.0 to 5.0-8.0 for its optimal activity which leads to a rise in operational cost. To address this issue, ENZbleach has been developed and optimized for pulp biobleaching without the need for pH adjustment of the pulp, which will make the large-scale operations more simple and cost effective. Moreover, ENZbleach is a cellulase-free xylanase, thus does not destroy the structure of cellulose and diminish pulp quality.
Annual Report 2011 48
Open Lab to Industry
pen Lab is a new activity initiated in 2011 to promote o Open Lab: Enzyme Technology Laboratory The laboratory’s Ocollaboration between laboratories and industries by activities include all aspects of enzyme biotechnology from inviting companies to meet with BIOTEC scientists, be informed screening of enzymes from microbial isolates, taking advantage of BIOTEC expertise and potential of the biotechnology to of Thailand’s biodiversity and from metagenomic libraries, make an improvement to industry, with expected results such gene isolation, enzyme production in wild-type microbes as collaborative research, commissioned research or analytical and recombinant systems to development of enzymatic service provision. Three laboratories were selected for three processes in industry. The Laboratory currently collaborates different events in 2011, based on the industrial application with several private companies to develop enzymes suitable of the technology: for each industry and process, for instance, alkaline-tolerant xylanase for pulp bleaching process in paper production, mixed enzymes for biofuel production, effective enzymes for o Open Lab: Food Biotechnology Research Unit The Food Biotechnology Research Unit focuses on research and animal feed and feed additives. development of food, with a special emphasis on using biotechnology to improve food quality to increase the value of o Open Lab: Genome Institute The Institute has built up commercial products. The research includes the development of capability for genome sequencing and analysis for plant and starter culture technology for fermented food, food chemistry, animal trait evaluation and improvement as well as identifying food safety and risk assessment. Several technologies have markers for breeding programs. Projects with industrial partners successfully been developed and are in use in industry, such include an identification of DNA markers for important traits for as starter culture technology for Thai traditional fermented a breeding program of eucalyptus, rubber tree and oil palm. pork sausage (Nham), acceleration of fish sauce fermentation with enzymes, fermentation process and a selected bacillus A total of 35 companies and venture capitalists participated strain to produce feed additives, production of pentosanase in the events, consisting of SMEs to major companies such from locally-isolated microorganisms for animal feed, and as CP Group, Saha Farms, Mitr Phol Sugar Group, Agromed, starter culture technology for sour soy sauce manufacture. Merck and Vnet Capital.
National Center for Genetic Engineering and Biotechnology (BIOTEC) 49 BIOTEC and Microinnovate Showcase Potential of Biotechnology Research to Industry
ommencing in 2006, BIOTEC has collaborated with SPM On 23 August 2011, BIOTEC and CFeed Co. Ltd. on a project to develop the fermentation Microinnovate co-organized process for Bacillus spp. to produce Bacillus-fermented an open-house event to product used as a feed additive. The trial was made in the 100 Microinnovate’s clients, SPM Group’s pig farms, resulting in a profit increase and composed of medium to large saving in imported feed additives. The success led to a joint farm operators in Thailand. investment between NSTDA and SPM, totaling 130 million The event aimed to inform baht, to set up a fermentation plant to produce feed additives farm operators of the R&D and microbial cells for commercial sale. The joint venture was behind the DS-1 product and named Microinnovate and the feed additive product is now biotechnology application to food and feed research. In marketed under the name “DS-1”. addition, guests also had a chance to visit BIOTEC state-of- the-art laboratories.
New TV Show Promoting Products and Technologies
n June 2011, NSTDA launched a new 1-hr TV show, Long ILaew Ruai, broadcasting on Sunday on Channel 9 public television. Each episode of Long Laew Ruai features a selected technology or product from NSTDA research activities that have been made available in the market. It details the research as well as the commercialization side, with participation of researchers, investors (in the case that the technology has been licensed to the private sector) and users of the technology. Five shows have been produced on BIOTEC technologies, namely bioremediation agent (based on research to screen for oil- degrading microorganisms found in Thailand), Aqua RASD (a closed recirculating aquaculture system integrated with hybrid nitrification biofilter tanks for ammonia removal and patented Tubular Denitrification Reactor for nitrate removal), flood- tolerant rice cultivar and biogas technology.
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HUMAN RESOURCES DEVELOPMENT
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IOTEC places high priority on capacity building through increasing Bthe quantity and quality of human resources in biotechnology as well as upgrading and educating the workforce. Several activities were designed to assist different segments of the workforce as well as address a variety of objectives, ranging from providing scholarships/fellowships to organizing training workshops for academics and industries, to organizing youth programs. In 2011, a total of 44 workshops were organized with over 2,000 participants attending in total. Eleven post- doctoral fellowships were granted, five of which are new fellowships approved in 2011. Workshops and science camps were organized, benefiting over 3,000 teachers and students.
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Capacity Building in Institutional Biosafety Committees (IBCs)
ith several years of biosafety experience, BIOTEC has BIOTEC continually facilitates the technical capacity of these Wbeen entrusted by the National Research Council IBCs through training and the dissemination of information. of Thailand (NRCT) to implement a project to build up the capacity of Institutional Biosafety Committees (IBC) in 2011. In order to strengthen capacity building to all 35 IBCs, NRCT provided financial support to BIOTEC to set up the 2011 training IBCs were established in various research and academic in human resource development for IBC network. With this institutes throughout Thailand to supervise the internal support, BIOTEC in collaboration with IBC’s nodes, developed research in laboratories and greenhouses to comply with training curriculum and organized practical hands-on training the Biosafety Guidelines for Research and Development, workshops for using the Guidelines to IBCs covering four developed by the Technical Biosafety Committee (TBC) with regions of Thailand. Four workshops were held in Chiang Mai, BIOTEC as its Secretariat. Currently, there are 35 IBCs scattered Khon Kaen, Bangkok and Songkhla provinces, and attended in four regions of Thailand. As the national coordinator of IBCs, by 180 officers.
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International Training Courses
hrough a strong network with international partners, o Training Workshop on Soil and Tissue Analysis for TBIOTEC organizes a number of international training courses Heavy Metals Organized on 21-22 February 2011, the catering to research and academic community in the region, Workshop with instructors from the Institute of Soil Science, with technical support from its international allies. In 2011, Chinese Academy of Sciences (ISSCAS) provided training on three international workshops were organized. soil standard making, monitoring of bioremediation process and regional soil pollution survey. The workshop drew about o Preservation and Management of Microbial Resources 30 attendees from the universities and government sector. Workshop The training workshop on Long-Term Preservation and Management of Microbial Resources with Agricultural o Regional Workshop on Gene Discovery from Uncultured Importance, held from 23-27 May 2011 at Thailand Science Microbes Using Metagenomic Approach The workshop, Park, was organized in cooperation with members of Asian held on 14 – 17 December 2010, aimed to provide knowledge Consortium for the Conservation and Sustainable Use and encourage researchers on utilization of uncultured of Microbial Resources (ACM) and World Data Centre for microbes for gene discovery that would serve as a prolific Microorganisms (WDCM). The training aims to increase the source for novel enzymes or effective bioactive compounds, capabilities of participants in the proper handling, long- and hand-on practice in handling and learning the laboratory term preservation and quality control of microorganisms to techniques relevant to unculturable microbes. Researchers maintain high quality inoculants which will help ensure their from BIOTEC and the National Institute of Advanced Industrial effectiveness in the field, including enhancing the awareness Science and Technology (AIST), Japan served as instructors. and appreciation on the benefits of high quality microbe- The Workshop was attended by twenty selected participants based fertilizers and bio-control products. Lectures and lab from seven different countries -- Indonesia, Lao PDR, Malaysia, practice were delivered by experts from China, Japan, Korea, Myanmar, Philippines, Thailand and Vietnam, ten of which the Philippines and Thailand. Nearly 70 participants made up were sponsored by funding from the Asia-Pacific International of researchers, lecturers and Ph.D. students from Cambodia, Molecular Biology Network (A-IMBN). Indonesia, Lao PDR, Malaysia, Myanmar, the Philippines, Vietnam and Thailand attended the Workshop.
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Training Program for Dairy Farmers on Steroid-based Synchronization of Bovine Ovulation
roblems on reproductive efficiency of dairy cows are Pprobably the most important problems for dairy producers across the globe. Controlling and synchronizing ovulation has become a routine reproductive management tool for dairy producers. BIOTEC Animal Physiology Laboratory has studied many hormonal treatment regimens, particularly steroid hormones in controlling ovulation. The steroid-based synchronization of ovulation protocol developed in the Laboratory has produced a good synchronization of follicular waves and sufficient synchronization of ovulation time, resulting in a useful protocol for timed artificial insemination (timed-AI). Field trials showed that the protocol can improve the conception rates in repeat-breeding dairy cows and also overall reproductive efficiencies in dairy cow herds. With good performance and cost-effectiveness, this protocol has been In order to popularize this technology among dairy farmers transferred to, and implemented in, a number of dairy co-ops across the country, BIOTEC and Pfizer (Thailand) implemented a and major dairy companies. training program on steroid-based synchronization of ovulation protocol in 2011. A total of eight workshops were organized in eight different provinces to give hands-on practical training to farmers and local livestock development government officials. The training program was attended by 380 individuals.
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Capacity Building Frontier Science in Food Processing Camp and Good Hygiene n 2011, the Ministry of Science and Technology launched a Practices for Rural Iproject “Frontier Science Camp”, aiming to upgrade science teachers in rural areas. Four camps were held -- one each in Communities Nakhon Sawan, Phitsanulok, Nakhon Ratchasima and Surat Thani provinces. Having experience designing teaching curriculum and materials in life science and also running its IOTEC works in collaboration with 14 local universities own Science in Rural Schools Program (SiRs), BIOTEC actively Bacross the country to provide technical assistance to participated in the project, alongside other research centers village communities to improve their local food product quality under NSTDA. The subject selected for training science teachers and safety. In 2011, the program assisted 23 communities, was enzyme technology. In total, the Project has trained over resulting in over 40 products from participating communities 600 science teachers in four regions of Thailand. to be awarded certificates from the Thailand Food and Drug Administration (FDA). Examples of products obtaining FDA certificates are: cassava cracker, macadamia nut products, mangosteen candy.
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PUBLIC AWARENESS
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triving to make Thailand a knowledge-based economy, one of SBIOTEC missions is to create public awareness of biotechnology, especially relating biotechnology and life sciences to everyday life. The mission is carried out by channeling information through various available popular media such as the internet and television, public events as well as children’s books.
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Book and Multimedia CD ROM on the Thai Amazing Microbes Rice College Exhibition
n 2002, PTT Exploration and Production PCL (PTTEP), in col- n 6 - 21 August 2011, the Ministry of Science and Ilaboration with the Population and Community Development OTechnology hosted the National Science and Technology Association (PDA) and the Thai Rice Foundation under Royal Fair 2011 at Bangkok International Trade & Exhibition Centre Patronage established the Thai Rice College Project in Kok (BITEC). The S&T Fair is organized with the aim to stimulate Klang, Buriram Province to improve the sustainability of rice public interest in science and technology and promote farming as well as act as continuing learning and development knowledge, understanding and dissemination of research center for Thai farmers. Training courses were developed to and development advances in science and technology for provide knowledge and technology on the improvement youth and the general public. of the quality of jasmine 105 rice, increase the productivity, reduce the costs of farming and reduce the use of chemical During the Fair, BIOTEC’s exhibition was organized under the fertilizers. The courses included the following lessons: farmland theme “Amazing Microbes” offering a close look into the micro- leveling, productivity improvement technology, integrated bial world through the images in a microscope and enlarged methods for the prevention and elimination of the insects models of microorganisms. Among the topics presented in and diseases, and harvest methods for quality and productivity the exhibition were: the discovery of the first bacteria; variet- improvement. As a result of this project, farmers in the area ies of microbial applications such as ethanol production and were able to apply the knowledge to their rice production, insulin production; as well as the collection and preservation becoming more self-sustained, gaining more income and of microorganisms to study their applications. improving their standard of living. The Thai Rice College at Kok Klang serves as a model for farmers in other regions and other organizations.
BIOTEC was commissioned by PTTEP to produce learning materials based on the training courses and also knowledge cultivated by local farmers through the training process. A book, accompanied by a multimedia CD ROM, was completed in late 2010. A total of 3,500 copies were produced for PTTEP to distribute to farmers.
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Participation in the 2011 National Dairy Fair
n 5-11 January 2011, the Dairy Farming Promotion As a part of research collaboration with DPO, BIOTEC and OOrganization of Thailand (DPO) organized 2011 National NSTDA participated in the exhibition to showcase a variety Dairy Fair at Muak Lek District, Saraburi Province. The opening of technologies as well as research and development in dairy ceremony on 5 January was graciously presided over by Her cows that could be utilized for reproductive and farm manage- Royal Highness Princess Maha Chakri Sirindhorn. The event ment. The exhibition represented the main R&D on steroid- featured the exhibitions of dairy farm industries, government based program for synchronizing ovulation, embryo transfer agencies and private sector. Also, the practical aspects of dairy technology, forage cane for animal feed and the development farming and several lectures from cattle experts in the fields of FrieBrah cattle management system. The BIOTEC research of dairy feeding, forage crop production, cattle breeding, team also provided a demonstration on embryo transfer treat- milk collection and processing, disease prevention and farm ment and management at the Fair. management were displayed and demonstrated to the visitors.
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Raising Awareness on Intellectual Property Rights
IOTEC, along with the Central Intellectual Property and BInternational Trade Court (IP&IT Court) and True Visions, joined by the British Council Thailand, initiated the Young Thai Documentary on IP Law Ambassador Award in 2006. The award aims to raise awareness of the legal and cultural issues regarding intellectual Public Television property (IP) among the younger generation, including creat- ing an informal network for university law students and legal ecognizing the power of television for reaching out to professionals. The competition is held every year, comprising millions of people in all walks of life, NSTDA produces an essay contest and a moot competition. BIOTEC plays an R documentaries for broadcasting via public television to teach important role in setting the topics for the essays and moot science as well as to demonstrate the results and impact of competition for each year and providing technical guidance inventions through NSTDA research. In 2011, BIOTEC intro- to the competition. duced the following topics in the documentary: Innovation
Technology for Phyto-remediation on Salinity Land, antibody On 23 November 2010, the 2010 Young Thai IP Law Ambassador array technology for disease detection in food and agriculture, Award Competition was launched, with an essay topic on “The biodiversity: importance and application, meat quality genetic Development of the Thai Intellectual Property System towards testing and PCR-based detection of tuberculosis. the ASEAN community” and the English moot competition on “Intellectual Property Dispute in Government Funded Research”. A total of 32 teams of law students from public and private universities entered the competition with the winning team, announced in April 2011, comprising two students from The Faculty of Law, Chulalongkorn University.
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Open Lab to Public
hailand Science Park (TSP), home of BIOTEC and its five Tresearch units, is considered the largest R&D community in Thailand with government-funded research labs and also private labs of several companies. TSP thus becomes a study site for schools, universities, government and private orga- nizations, as well as the general public. BIOTEC realizes this opportunity to raise awareness of biotechnology, especially the importance and relevance of technology to everyday life Numbers of visitors to BIOTEC Lab Tour and the economy, and therefore has an open-door policy to welcome the public to have a tour of the laboratory facilities on a regular basis, with close interaction with BIOTEC research staff. This interaction also encourages research staff to relate their research to problem solving with the public. International Groups 53 130 Domestic Groups
In 2011, there were a total of 183 groups touring BIOTEC laboratories at Thailand Science Park. Examples of visitors include: Department of Trade Negotiations (Thailand), Minister 85 of Science and Technology (Thailand), science teachers from 19 20 Chantaburi Province (Thailand), Ajinomoto (Thailand), Lee Feed Mill (Thailand), Institute of Forensic Training and Research 14 (Thailand), Japanese Chamber of Commerce (Thailand), Pfizer, Chinese Academy of Sciences, Vietnamese Ministry of 24 Science and Technology, high school students from Korea and 21 Singapore, Biotechnology Industry Study Group of National Defense University (USA) and Bogor Agricultural University (Indonesia). Academic institute Government organization Private company
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INTERNATIONAL COLLABORATION
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he BIOTEC International Cooperation Program aims to capitalize Ton international linkages to help BIOTEC and Thailand become a regional leader in the field of biotechnology. In so doing, the Center has developed close linkages with overseas organizations at the bilateral, multilateral and regional levels. The goal is achieved through activities such as establishing formal collaborative agreements, organizing joint scientific seminars with international partners as well as establishing and organizing an annual meeting of the BIOTEC International Advisory Board. In 2011, BIOTEC entered into MOU’s to foster collaboration with the following organizations: Mie University (Japan), Institute of Medical Science of University of Tokyo (Japan), World Data Centre for Microorganisms (China) and the Academy of Sciences for the Developing World (TWAS).
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UK-Thailand Partners in Science
nder the ‘UK-Thailand Partners in Science’ program, Uformalised in September 2009 by the UK Government Chief Scientific Adviser and the National Science and Technology Development Agency (NSTDA) of Thailand, BIOTEC, to collaborate with the University of Liverpool. The BIOTEC and the British High Commission in Singapore, jointly network on natural products has also reached an agreement identified three areas of potential collaboration: foodborne on one joint project on a type of insect pathogenic fungi, pathogens, natural products and genomic technology. Ophiocordyceps unilateralis, with the topics of marine fungi To initiate this collaboration, a series of workshops and and actinomycetes still being explored. Ophiocordyceps meetings were organized throughout the first half of the unilateralis project is to be funded by BIOTEC, and will consist 2011 fiscal year, bringing in UK scientists to meet with their of partners including BIOTEC, the Genome Analysis Centre Thai counterparts on each specific theme. These meetings (TGAC) of the John Innes Centre, and the University of Bristol. resulted in the formation of two UK-Thailand networks, one In addition to research, both networks of natural products on foodborne pathogens and the other on natural products. and foodborne pathogens also include a capacity building The UK-Thailand network on foodborne pathogens currently component, through the provision of PhD scholarships for has one research project on microbial risk assessment of Thai students to be jointly funded by Thailand Research Campylobacter in the broiler chicken production chain in Fund and the UK’s Biotechnology and Biological Sciences Thailand. This was funded by NSTDA which provided grants Research Council (BBSRC). to Chulalongkorn University, Khon Kaen University, King Mongkut’s University of Technology Thonburi (KMUTT) and
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Joint Research on Antibody Array Technology for Multiplex Detection of Plant Diseases
etection and identification of plant pathogens is essential an additional sandwich PhD scholarship jointly funded by Dto epidemiological and disease management studies, as BIOTEC and QUB. Combining QUB’s expertise in SPR and well as to selective breeding programs for pathogen-resistant assay development with BIOTEC’s competence in microarray plants and certification of disease-free seeds for import and technology and unique collection of highly specific antibodies, export. A joint project to develop multiplex detection of the project aims to develop two antibody-based diagnostic plant diseases in melon using antibody array technology methods, ie. a multiwell-plate antibody array and multiplex and multiplex SPR was conceived between scientists from SPR, to simultaneously detect multiple seed-borne pathogens. BIOTEC and the Institute of Agri-Food & Land Use, Queen’s Although the methods developed in this particular project University Belfast (QUB). The three-year research project, target at melon diseases, they can serve as prototypes for commenced in December 2010, is funded by NSTDA, with other different diagnostic purposes.
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BIOTEC – ETH Fostering Workshop on Collaboration on Functional Genomics Phyto-bioremediation in Rice and Cassava with China
o establish research collaboration with the Swiss Federal n 20 – 25 February 2011, BIOTEC hosted a visit of research TInstitute of Technology Zurich (ETH), BIOTEC/NSTDA invited Oscientists from the Institute of Soil Science, Chinese scientists from ETH Zurich Department of Biology, led by Prof. Academy of Sciences (ISSCAS), China. As part of the visit, Wilhelm Griussem, to Thailand to organize the BIOTEC – ETH BIOTEC and ISSCAS co-organized a Training Workshop on Soil Workshop on Functional Genomics and Biotechnology Studies and Tissue Analysis for Heavy Metals, with Chinese scientists in Rice and Cassava on 6-8 July 2011. The Workshop aimed sharing their knowledge and experience in soil standard to promote research collaboration between Thai and leading making, monitoring of bioremediation process and regional ETH scientists, encourage knowledge exchange and explore soil pollution survey. In addition, a roundtable discussion areas of common interest in functional genomics and systems was held to explore and discuss the scope and challenges biology in rice and cassava. Attending the Workshop were of possible research collaboration on phyto-bioremediation various research groups from Thailand, including BIOTEC (starch technologies for heavy metals. The meeting brought together biosynthesis, cassava genome and rice molecular breeding), representatives of Chulalongkorn University, King Mongkut’s Kasetsart University (rice functional genomics), King Mongkut’s University of Technology Thonburi and BIOTEC. The mutual University of Technology Thonburi (systems biology of cassava research interests included phytoremediation of heavy metal starch synthesis) and Chulalongkorn University (salt tolerance contamination in agricultural soil focusing on cadmium mechanism in rice). remediation in paddy rice field, risk assessment and risk based soil standards, management of acid mine drainage in mining areas, bioremediation of organic pollutants: mechanisms and application and R&D of innovative physiochemical remediation technologies.
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Collaboration with Short Course for World Data Centre Students from for Microorganisms Soon Chun Hyang (WDCM) University, Korea
n 14 September 2011, an MOU signing ceremony between ecognized for research competency and industry-oriented OBIOTEC and the World Data Centre for Microorganisms Rresearch programs, BIOTEC was entrusted by Soon (WDCM) was held with the purpose to facilitate mutual access Chun Hyang University (SCH) in Korea to organize a 3-week to the biological resources and information, promote joint course for undergraduate students from Faculties of Science, research and exchange researchers. The MOU signing was Medical Science and Medicine in January 2011. Twenty SCH conducted in the presence of Dr. Philippe Desmeth, President students attended the course, which includes a 5-day lecture of World Federation for Culture Collections (WFCC) and Dr. class – a broad overview of the biotechnology field including Juncai Ma, Director of World Data Centre for Microorganisms Bioresources Diversity and Utilization, Food Biotechnology, (WDCM), whose visit was to provide advice to BIOTEC on Sensors and Diagnostics and Plant Science- and hands-on making the transition from culture collection to a Biological research practice in individual laboratories. The program also Resource Centre (BRC). offered opportunities for students to visit a food-processing factory and a Biopharmaceutical cGMP Pilot Plant. These BIOTEC Culture Collection (BCC) was established in 1996 to activities will provide SCH students with a well-rounded serve as a central facility to collect and maintain microor- academic experience in Thailand. ganisms and their relevant data, as well as a repository of patent-related microorganisms of Thailand Department of Intellectual Property. BIOTEC also houses a storage facility for other biomaterials such as hybridoma clones. To fully serve as a source of all bioresource essentials, including services for research and development in life sciences, BIOTEC is currently in the process of establishing a BRC.
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Human Resource Development Program in Biotechnology
n July 2011, BIOTEC welcomed twelve researchers from Ithe Asia-Pacific region to attend the Human Resource Development Program in Biotechnology, a program to sponsor young researchers from the region for on-the-job training in BIOTEC laboratories for 3-6 months. This year participants are young researchers from seven countries, namely Fiji, Indonesia, Malaysia, Myanmar, Papua New Guinea, the Philippines and Vietnam. The Program aims to build up R&D capability for developing countries as well as foster research networks among countries in this region.
As this Program places importance on improving research In addition to lab training, participants had an opportunity to skills, it is designed to be an on-the-job and research-based visit factories or other research institutes to gain experience in training in BIOTEC’s laboratories under the guidance of BIOTEC biotechnology and bioindustry development in Thailand. At researchers. The program of training is divided into two seg- the end of the training, all trainees gave seminars of their ments, fundamental and advanced courses. During a period research work to their peers and BIOTEC researchers. of three months, ten researchers in the fundamental course were trained in basic techniques in different research areas Commencing in 2001, this program has sponsored 124 fellow- such as plant biotechnology, protein-ligand engineering and ships, with successful outcomes measured by joint publications molecular biology, shrimp molecular biology and biotech- between BIOTEC and foreign researchers, scholarships acquired nology, biodiversity and bioresource utilization, while two by alumni for higher education in developed countries and other participants stayed over a period of six months to learn career progression in home countries. advanced techniques.
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Visiting Researchers
IOTEC regularly has foreign researchers spending time in BBIOTEC laboratories. Some visiting researchers come to work Internship Program on existing research collaboration, some arrive with a program aiming to initiate and foster collaboration such as BIOTEC Visiting for Foreign Students Scientist Scheme and TWAS-UNESCO Associateship Scheme, and some make a request for training on specific topics by BIOTEC researchers. In 2011, sixteen foreign researchers and very year, BIOTEC welcomes foreign students to join BIOTEC experts joined BIOTEC, from Nigeria, Sweden, the Philippines, Elaboratories to gain research experience. Agreements have Indonesia, Malaysia, India, Bangladesh and USA. been reached with Atma Jaya Catholic University (Indonesia); University of Kent (UK), University of Liverpool (UK), National Amongst the researchers spending time at BIOTEC was Dr. University of Taiwan (Taiwan), Temasek Polytechnic (Singapore) David Matthews who was a visiting scientist from the US. An and Nanyang Polytechnic (Singapore) to send students to expert on biomolecular structures and drug discovery, Dr. BIOTEC on a regular basis. In addition, direct and individual Matthews was invited to BIOTEC for one month to review and applications from students are also welcomed. In 2011 fiscal advise the BIOTEC research team on potential drugs targeting year, BIOTEC hosted over fifty students from regular institutes malarial dihydrofolate reductase antifolate. During his visit, as well as from University of British Columbia (Canada) and Dr. Matthews also shared his expertise with the Thai research University of Montpellier 2 (France). community through two public seminars.
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IMPACT OF BIOTEC’S OUTPUT
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IOTEC constantly monitors and assesses the impact of its output Btowards social and economic development. Every year, a number of technology transfer projects are selected for detailed impact study for the following few years. The impact is measured in the form of income generated by the clients out of such products and technologies, and also quantified from parameters such as import substitution and employment generation.
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Agriculture and Food Cluster
echnology transfer of rice seed production to farmers: households were selected to join the program, with cultivation TThanyasirin Thanyasirin is the RD6 cultivar, a jasmine area of 205 rai (33 hectares). 112 tonnes of seed were produced glutinous rice, with blast resistance added into its genome using in 2009, worth 2 million Baht at the market price of 18 Baht/ molecular markers in the backcross breeding. Thanyasirin has kg. 65 tonnes of 2009 seed were planted in 2010/2011, and high cooking and eating qualities. It shows drastic improvement is expected to yield 6,500 tonnes of grains. At the price of 10 in leaf and neck blast resistance to a broad range of spectrum Baht/kg, the grain to be produced is worth 65 million Baht, in the rainfed lowland areas. Seed production of Thanyasirin bringing the total income generated from the technology was transferred to four northern provinces (Nan, Chiang Rai, to 67 million Baht. Lampang and Chiang Mai) through collaboration between BIOTEC, NSTDA, Rajamangala University of Technology Lanna Technology transfer of rice seed production to farmers: and local authorities. In the 2010/2011 growing season, 260 Homcholasit Homcholasit is an irrigated rice (non photope- households participated in the program, with the cultivation riod sensitivity) with aromatic and good eating quality. It is area of 753 rai (120.5 hectares). 554 tonnes of seed were derived from a cross between KDML105 and IR57514 and was produced, valued at 14 million Baht (at 25 Baht/kg). This developed for flash flood tolerance. In 2010, BIOTEC, NSTDA volume of seed can further produce 40,791 tonnes of grain, and Phakhai Agricultural Cooperative implemented the tech- worth 408 million Baht. The technology has generated 422 nology transfer of Homcholasit seed production in Ayuthaya million Baht in total along the value chain. The technology Province. The program was joined by 15 households, with 125 transfer has also been implemented in Chaiyaphum Province, rai of land (20 hectares). 44.6 tonnes of seed were produced, in collaboration with the Chumphae Rice Research Center. 48 generating 3 million Baht in income from selling seeds and
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grains. During off-season crop production (December 2010 – Establishment of disease-free sugarcane production unit April 2011), the program expanded to more cooperatives in BIOTEC provided technical assistance to Mitr Lao Sugar Co., four provinces (Ayuthaya, Chainat, Ang Thong and Uttaradit), Ltd. to establish a disease-free sugarcane production unit, with the participation of 93 households. 450 tonnes were the largest unit in Asia, to serve as a stock parent plant for harvested from 965 rai of land (154 hectares). 178 tonnes the company’s sustainable production. In 2010/2111, 5200 met the standard and were sold as seed, fetching 3.9 million tonnes of cane stem were produced, worth 12 million Baht. Baht. This amount of seed can be planted in 8,900 rai (1,424 hectares) and is expected to produce 5,340 tonnes of grain, Establishment of microbiology laboratory BIOTEC provided worth 42.7 million Baht. 272 tonnes of sub-standard seed are technical assistance to Mitr Phol Research and Development sold as grain, worth 2.7 million Baht. In total, the technology Center Co., Ltd., to set up a Microbiology Laboratory, resulting can generate 43.9 million Baht. in an investment of 10.7 million Baht in scientific equipment and construction of microbial production plant, and an ad- Pentosanase production technology for animal feed ditional 1 million Baht of employment/year. Microorganisms industry BIOTEC has developed the technology, which in- produced have replaced 1.5 million Baht worth of imports. The cludes the pentosanase-producing microorganism carefully use of these microorganisms reduced production costs and screened from BIOTEC Culture Collection and the production increased ethanol production, estimated at 6.2 million Baht. process to achieve a product in a powder form. The technol- ogy was licensed to Asia Star Animal Health Co., Ltd. (ASAH) to Rooftop plant cultivation technology SCG Building Materials manufacture and distribute pentosanase as feed additives. In commissioned BIOTEC to screen for suitable plants and to 2011, the company earned 4.9 million baht in sales from the develop technology for rooftop plant cultivation in order to product, saving the country 9.8 million baht from importing add value to SCG’s building materials. This technology en- 24.6 tonnes of this enzyme. The impact resulted in healthier abled SCG to add a “Roof Garden” system to its product line, animals, better meat quality and the saving on feed is estimated generating the 1st year income of 3.5 million Baht. to be 45.9 million Baht. Forage cane for animal feed BIOTEC, in collaboration with Bacillus subtilis and fermentation process to produce Kasetsart and Khon Kaen universities, studied the potential of feed additives The technology, which includes microbial using cane as forage for farm animals. In 2011, forage cane strain and fermentation process to produce feed additives, was planted in 50 rai (8 hectares) of Namfon Farm Co., Ltd. was licensed to Microinnovate Co., Ltd. (previously known and 23 rai (3.7 hectares) in Lopburi, Sa Kaew, Loei and Khon as SPM Science). In 2011, the company was able to produce Kaen provinces, producing 2.3 million Baht worth of cane. 170,280 Baht worth of culture, which was subsequently used to produce dry supplement product worth 12.6 million Baht. Monoclonal and polyclonal antibodies for ELISA test The use of feed supplement enabled farms to increase their BIOTEC collaborated with Kasetsart University in developing profits by 811,750 Baht. antibodies for ELISA tests of geminiviruses, tospovirus and potyviruses, whereas antibody for ELISA test of Acidovorax Embryo transfer technology BIOTEC, in collaboration with avenae subsp. Citrulli (Aac) was developed by BIOTEC, Khon Suranaree University of Technology and Namfon Farm Co., Kaen and Kasertsart universities. Disease detection made by Ltd., conducted a study on the performance of embryo these antibody assays enables seed farmers to produce high- transfer technology (ET). Cows produced from ET possessed quality seed. This has been estimated to accrue 27 million Baht better genetic parameters, with improved milk production in income to farmers and 180 million Baht worth of exports of 36,000 Baht/lactation/cow. The project has generated 60 to seed companies. cows, producing 2.1 million Baht worth of fresh milk.
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NPV production for insect pest control BIOTEC established a Nuclear Polyhedrosis Virus Pilot Plant for Insect Pest Control. In 2011, BIOTEC earned 2.1 million Baht in sales from the NPV product. The use of NPV can reduce chemical insecticide cost by 1.8 million Baht. The product also saved the country 5.7 million baht from importing biocontrol agents. The indirect impact, quantified from the safety of the product (as opposed to chemical pesticides) to human health and environment, is estimated to be 2.8 million Baht.
Agro-industry Practice School Program BIOTEC/NSTDA supported the establishment of Agro-industry Practice School Program at Prince of Songkhla University (PSU) in 2007. The Practice School Program is an intensive graduate course plac- ing strong emphasis on practical experience at industrial sites to give students hands-on experience. Since its inception, Agro-industry Practice School Program attracted 23 factories to participate as a practice site, produced 17 graduates, and generated 48 projects with industry. In 2011, the projects resulted in an estimated savings of 7 million Bath through improvements to factory efficiency, reduction of production losses, reduction of waste, for instance.
Health and Medicine Alpha thalassemia immunochromatographic strip test The technology was licensed to i+MED Laboratories Co., Ltd. Cluster In 2011, i+MED Laboratories earned revenue of 610,911 Baht in sales. This product was able to save the cost of detection by 7.7 million Baht compared to other available methods. A iagnostic test for influenza BIOTEC collaborated with blood test to check for thalassemia is important, especially for Innova Biotechnology Co., Ltd., one of the NSTDA joint D couples planning to have children. If doctors find that either venture companies, in developing an immuno-chromatographic parent is a carrier, they will recommend the couple avoid test kit for influenza type A and subsequently licensing this pregnancy and this helps early screening of children who technology to Innova for commercial production. In 2011, would be prone to thalassemia. Therefore, indirect impact Innova earned revenue of 255,534 Baht in sales of “Innova Flu-A” of 21.5 million Baht in healthcare saving is estimated for the product. This product was able to save the country 766,602 prevention of thalassemia transmission. Baht from the importation of influenza kits. The availability of this rapid test kit enabled the effective control of the disease, estimated to be 5.6 million Baht in healthcare savings from the spread of the disease. Energy and
Clove and cinnamon oil formulation for dust mite fumiga- Environment Cluster tion The technology, developed by King Mongkut’s Institute of Technology Ladkrabang (KMITL), was licensed by BIOTEC tandard test for determining the resistance of exterior and KMITL to Good Guy Group Co., Ltd. and Natural Herb emulsion paint to algae The protocol for standard test to Product (Thailand) Co., Ltd. for commercialization in 2009. In S determine the resistance of exterior emulsion paint to algae 2011, the product generated 3.5 million Baht in revenue. The was developed by scientists from the Thailand Institute of product is estimated to create an indirect impact of 1 million Scientific and Technological Research (TISTR), with BIOTEC Baht in preventing allergies caused by dust mite. research grant. TISTR later acquired the license to offer the
National Center for Genetic Engineering and Biotechnology (BIOTEC) 75
service based on this protocol. In 2011, the testing service Capacity building on energy and resource efficiency for generated 150,000 Baht in revenue, saving the country 1.4 Thai native starch industry In 2009, BIOTEC/NSTDA and GTZ million Baht from sending the paint to be tested in foreign launched the project aiming to increase the competitiveness countries. of Thai native tapioca starch industry through practical train- ing program for, and providing advise to, starch factories Promotion of biogas technology in cassava starch industry to make improvements on efficient production processes, The biogas technology, for wastewater treatment and methane pollution control mechanisms, waste recycling/re-use, and production, developed by EcoWaste has been implemented energy efficiency options. For the 14 factories participating in 4 factories, resulting in an average of 116 million Baht in in the project, a total of 40 million Baht in savings was esti- energy saving annually. In 2011, two more factories have mated from waste minimization and making improvements signed up for the technology transfer, with an investment of to the machinery. 80 million Baht in total. Innovation technology for phyto-remediation on salinity Promotion of biogas technology in agri-food industry land BIOTEC in collaboration with SCG and several organiza- The biogas technology was installed in 3 agri-food factories. tions implemented the technology for phyto-remediation on The value of waste treatment cost saving, energy saving and salinity land. In 2011, over 1100 households in Sakon Nakhon electricity generation totaled 31 million Baht. 44 million Baht and Udorn Thani provinces joined the project, covering the of additional investment was made in 2011. area of 5000 rai (800 hectares). The project was able to turn arid land into being arable for rice and other crops, selected Promotion of biogas technology in palm oil industry for the area. Farmers were able to subsist on the rice and farm Research and engineering team at EcoWaste has designed produce, estimated to be worth 16 million Baht. and installed a hybrid reactor to treat wastewater from palm oil production in one factory. The technology generated 7.4 million Baht worth of energy and 50 million Baht of additional investment.
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77
Appendices
List of Publications
1. Adachi, O., Hours, R.A., Akakabe, Y., Tanasupawat, S., 4. Aewsiri, T., Benjakul, S., Visessanguan, W., Wierenga, Yukphan, P., Shinagawa, E., Yakushi, T. and Matsushita, K. P.A. and Gruppen, H. (2011). Emulsifying Property and (2010). Production of 4-Keto-D-arabonate by Oxidative Antioxidative Activity of Cuttlefish Skin Gelatin Modified Fermentation with Newly Isolated Gluconacetobacter with Oxidized Linoleic Acid and Oxidized Tannic Acid. liquefaciens. Bioscience Biotechnology and Biochemistry, Food and Bioprocess Technology, doi: 10.1007/s11947- 74(12), 2555-2558. 011-0636-1. 2. Aderibigbe E. Y., Visessanguan W., Sumpavapol P. and 5. Aewsiri, T., Benjakul, S., Visessanguan, W., Wierenga, Kongtong K. (2011). Sourcing starter cultures for Parkia P.A. and Gruppen, H. (2011). Improvement of foaming biglobosa fermentation I: Phylogenic grouping of Bacillus properties of cuttlefish skin gelatin by modification species from commercial ‘iru’ samples. International with N-hydroxysuccinimide esters of fatty acid. Food Journal for Biotechnology and Molecular Biology Research, Hydrocolloids, 25(5), 1277-1284. 2(7), 121-127. 6. Aewsiri, T., Benjakul, S., Visessanguan, W., Wierenga, P.A. 3. Aewsiri, T., Benjakul, S., Visessanguan, W., Encarnacion, and Gruppen, H. (2011). Surface activity and molecular A.B., Wierenga, P.A. and Gruppen, H. (2011). Enhancement characteristics of cuttlefish skin gelatin modified by of Emulsifying Properties of Cuttlefish Skin Gelatin by oxidized linoleic acid. International Journal of Biological Modification withN -hydroxysuccinimide Esters of Fatty Macromolecules, 48(4), 650-660. Acids. Food and Bioprocess Technology, doi:10.1007/ 7. Aichayawanich, S., Nopharatana, M., Nopharatana, A. s11947-011-0553-3. and Songkasiri, W. (2011). Agglomeration mechanisms of cassava starch during pneumatic conveying drying. Carbohydrate Polymers, 84(1), 292-298.
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8. Alias, S.A., Zainuddin, N. and Jones, E.B.G. (2010). 17. Boonlarppradab, C., Suriyachadkun, C., Suphothina, S. Biodiversity of marine fungi in Malaysian mangroves. and Tobwor, P. (2011). Bireticulol, a bioactive isocou- Botanica Marina, 53(6), 545-554. marin dimer from Streptomyces sp. BCC24731. Journal 9. Amnuaykanjanasin, A., Panchanawaporn, S., Chutrakul, of Antibiotics, 64(3), 267-270. C. and Tanticharoen, M. (2011). Genes differentially 18. Boonyuen, N., Chuaseeharonnachai, C., Suetrong, expressed under naphthoquinone-producing condi- S., Sri-indrasutdhi, V., Sivichai, S., Jones, E.B.G. and tions in the entomopathogenic fungus Ophiocordyceps Pang, L.K . (2011). Savoryellales (Hypocreomycetidae, unilateralis. Canadian Journal of Microbiology, 57(8), Sordariomycetes): a novel lineage of aquatic ascomy- 680-692. cetes inferred from multiple-gene phylogenies of the 10. Amparyup, P., Klinbunga, S. and Jarayabhand, P. (2010). genera Ascotaiwania, Ascothailandia, and Savoryella. Identification and Expression Analysis of Sex-Specific Mycologia, doi:10.3852/11-102. Expression Markers of Thai Abalone Haliotis asinina, 19. Brück, T., Beaudry, C., Hilgenkamp, H., Karoonuthaisiri, Linneaus, 1758. Journal of Shellfish Research, 29(3), 765- N., Mohamed, H.S.-E., Weiss, G.A. (2010). Empowering 773. young scientists. Science, 328(5974), 17. 11. Aporntewan, C., Phokaew, C., Piriyapongsa, J., Ngamphiw, 20. Brück, T., Beaudry, C., Hilgenkamp, H., Kassen, R., C., Ittiwut, C., Tongsima, S. and Mutirangura, A. (2011). Karoonuthaisiri, N., El Din Mohamed, H.S., Weiss, G.A. Hypomethylation of Intragenic LINE-1 Represses (2010). Response--The Time of Young Scientists. Science, Transcription in Cancer Cells through AGO2. PLoS ONE, 329(5992), 626-627. 6(3), e17934. 21. Buatong, J., Phongpaichit, S., Rukachaisirikul, V. and 12. Arunrut, N., Phromjai, J., Gangnonngiw, W., Kanthong, Sakayaroj, J. (2011). Antimicrobial activity of crude N., Sriurairatana, S. and Kiatpathomchai, W. (2011). In extracts from mangrove fungal endophytes. World vitro cultivation of shrimp Taura syndrome virus (TSV) Journal of Microbiology and Biotechnology, doi:10.1007/ in a C6/36 mosquito cell line. Journal of Fish Diseases, s11274-011-0765-8. doi:10.1111/j.1365-2761.2011.01286.x. 22. Bunyapaiboonsri, T., Yoiprommarat, S., Srisanoh, U., 13. Arunrut, N., Prombun, P., Saksmerprome, V., Flegel, T.W. Choowong, W., Tasanathai, K., Hywel-Jones, N.L., Luangsa- and Kiatpathomchai, W. (2011). Rapid and sensitive ard, J.J. and Isaka, M. (2011). Isariotins G-J from cultures detection of infectious hypodermal and hematopoietic of the Lepidoptera pathogenic fungus Isaria tenuipes. necrosis virus by loop-mediated isothermal amplifica- Phytochemistry Letters, doi:10.1016/j.phytol.2011.04.018. tion combined with a lateral flow dipstick. Journal of 23. Busayarat, N., Senapin, S., Tonganunt, M., Phiwsaiya, K., Virological Methods, 171(1), 21-25. Meemetta, W., Unajak, S., Jitrapakdee, S., Lo, C.-F. and 14. Arunrut, N., Seetang-Nun, Y., Phromjai, J., Panphut, W. Phongdara, A. (2011). Shrimp laminin receptor binds and Kiatpathomchai, W. (2011). Rapid and sensitive with capsid proteins of two additional shrimp RNA detection of Laem-Singh virus by reverse transcription viruses YHV and IMNV. Fish and Shellfish Immunology, loop-mediated isothermal amplification combined with 31(1), 66-72. a lateral flow dipstick. Journal of Virological Methods, 24. Chaiyapechara,S., Rungrassamee, W., Suriyachay, I., doi:10.1016/j.jviromet.2011.06.020. Kuncharin, Y., Klanchui, A., Karoonuthaisiri, N. and 15. Asasutjarit, R., Thanasanchokpibull, S., Fuongfuchat, A. Jiravanichpaisal, P. (2011). Bacterial Community and Veeranondha, S. (2011). Optimization and evaluation Associated with the Intestinal Tract of P. monodon of thermoresponsive diclofenac sodium ophthalmic in in Commercial Farms. Microbial Ecology, doi:10.1007/ situ gels. International Journal of Pharmaceutics, 411(1-2), s00248-011-9936-2. 128-135. 25. Chantarasuwan, C., Benjakul, S. and Visessanguan, W. 16. Auephanwiriyakul, S., Sumonphan, E., Theera-Umpon, (2011). The effects of sodium bicarbonate on confor- N. and Tayapiwatana, C. (2011). Automatic Nevirapine mational changes of natural actomyosin from Pacific concentration interpretation system using support white shrimp (Litopenaeus vannamei). Food Chemistry, vector regression. Computer Methods and Programs in 129(4), 1636-1643. Biomedicine, 101(3), 271-281.
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26. Chantasingh, D., Kitikhun, S., Eurwilaichitr, L., 36. Cha-um, S., Yooyongwech, S. and Supaibulwatana, K. Uengwetwanit, T. and Pootanakit, K. (2011). Functional (2010). Water deficit stress in the reproductive stage expression in Beauveria bassiana of a chitinase gene from of four indica rice (Oryza sativa L.) genotypes. Pakistan Ophiocordyceps unilateralis, an ant-pathogenic fungus. Journal of Botany, 42(5), 3387-3398. Biocontrol Science and Technology, 21(6), 677-686. 37. Cheawchanlertfa, P., Cheevadhanarak, S., Tanticharoen, 27. Cha-um, C. and Kirdmanee, C. (2010). Acclimatisation M., Maresca, B. and Laoteng, K. (2011). Up-regulated ex- to mannitol-induced water deficit in miniature rose pression of desaturase genes of Mucor rouxii in response (Rosa x hybrida) plantlets cultivated in vitro under to low temperature associates with pre-existing cellular autotrophic conditions. Journal of Horticultural Science fatty acid constituents. Molecular Biology Reports, 38(5), and Biotechnology, 85(6), 533-538. 3455-3462. 28. Cha-um, C. and Kirdmanee, C. (2010). Effect of inland 38. Chotineeranat, S., Wansuksri, R., Piyachomkwan, K., salt-affected soil on physiological and growth charac- Chatakanonda, P., Weerathaworn, P. and Sriroth, K. ters of Eucalypt tree (Eucalyptus camaldulensis Dehnh.). (2010). Effect of calcium ions on ethanol production from Emirates Journal of Food and Agriculture, 22(6), 466-474. molasses by Saccharomyces cerevisiae. Sugar Technology, 29. Cha-um, C., Takabe, T. and Kirdmanee, C. (2010). Osmotic 12(2), 120-124. potential, photosynthetic abilities and growth characters 39. Chunglok, W., Wuragil, D.K., Oaew, S., Somasundrum, M. of oil palm (Elaeis guineensis Jacq.) seedlings in responses and Surareungchai, W. (2011). Immunoassay based on to polyethylene glycol-induced water deficit. African carbon nanotubes-enhanced ELISA for Salmonella en- Journal of Biotechnology, 9(39), 6509-6516. terica serovar Typhimurium. Biosensors and Bioelectronics, 30. Cha-um, C., Ulziibat, B. and Kirdmanee, C. (2010). Effects 26(8), 3584-3589. of temperature and relative humidity during in vitro 40. Chunglok,W., Khownarumit, P., Rijiravanich, P., acclimatization, on physiological changes and growth Somasundrum, M. and Surareungchai, W. (2011). characters of Phalaenopsis adapted to in vivo. Australian Electrochemical immunoassay platform for high sensitiv- Journal of Crop Science, 4(9), 750-756. ity protein detection based on redox-modified carbon 31. Cha-um, S. and Kirdmanee, C. (2011). In Vitro flowering of nanotube labels. Analyst, 136, 2969-2974. miniature roses (Rosa×hybrida L. 'Red Imp') in response 41. Daorueang, D., Thuwajit, P., Roitrakul, S., Laha, T., to salt stress. European Journal of Horticultural Science, Kaewkes, S., Endo, Y. and Thuwajit, C. (2011). Secreted 76(1), 239-245. Opisthorchis viverrini glutathione S-transferase regulates 32. Cha-um, S. and Kirdmanee, C. (2011). Photosynthetic cell proliferation through AKT and ERK pathways in chol- Features, Growth and Yield of Thai Jasmine Rice as angiocarcinoma. Parasitology International, doi:10.1016/j. Influenced by Saline Soil Treated with Organic Matter. parint.2011.07.011. Philippine Agricultural Scientist, 94(1), 23-28. 42. Dedsuksophon, W., Faungnawakij, K., Champreda, V. 33. Cha-um, S. and Kirdmanee, C. (2011). Remediation of and Laosiripojana, N. (2011). Hydrolysis/dehydration/ salt-affected soil by the addition of organic matter - an aldol-condensation/hydrogenation of lignocellulosic investigation into improving glutinous rice productivity. biomass and biomass-derived carbohydrates in the
Scientia Agricola, 68(4), 406-410. presence of Pd/WO3–ZrO2 in a single reactor. Bioresource 34. Cha-um, S., Chanseetis, C., Chintakovid, W., Pichakum, A. Technology, 102(2), 2040-2046. and Supaibulwatana, K. (2011). Promoting root induc- 43. Deo, R.P., Songkasiri, W., Rittmann, B.E. and Reed, D.T. tion and growth of in vitro macadamia (Macadamia (2010). Surface Complexation of Neptunium(V) onto tetraphylla L. ‘Keaau’) plantlets using CO2-enriched Whole Cells and Cell Components of Shewanella alga: photoautotrophic conditions. Plant Cell Tissue and Organ Modeling and Experimental Study. Environmental Science Culture, 106(3), 435-444. and Technology, 44(13), 4930-4935. 35. Cha-um, S., Pokasombat, Y. and Kirdmanee, C. (2011). 44. Duangchinda, T., Dejnirattisai, W., Vasanawathana, Remediation of salt-affected soil by gypsum and farm- S., Limpitikul, W., Tangthawornchaikul, N., Malasit, yard manure - Importance for the production of Jasmine P., Mongkolsapaya, J. and Screatona, G. (2010). rice. Australian Journal of Crop Science, 5(4), 458-465. Immunodominant T-cell responses to dengue virus NS3 are associated with DHF. Proceedings of the National Academy of Sciences of the United States of America, 107(39), 16922-16927.
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45. Faksri, K., Drobniewski, F., Nikolayevskyy, V., Brown, T., 55. Isaka, M., Chinthanom, P., Supothina, S., Tobwor, P. and Prammananan, T., Palittapongarnpim, P., Prayoonwiwat, Hywel-Jones, N.L. (2010). Pyridone and Tetramic Acid N. and Chaiprasert, A. (2011). Genetic diversity of the Alkaloids from the Spider Pathogenic Fungus Torrubiella Mycobacterium tuberculosis Beijing family based on sp. BCC 2165. Journal of Natural Products, 73(12), 2057- IS6110, SNP, LSP and VNTR profiles from Thailand. 2060. Infection Genetics and Evolution, 11(5), 1142-1149. 56. Isaka, M., Palasarn, S., Prathumpai, W. and Laksanacharoen, 46. Gutka, H.J., Rukseree, K., Wheeler, P.R., Franzblau, S.G. and P. (2011). Pimarane Diterpenes from the Endophytic Movahedzadeh, F. (2011). glpX Gene of Mycobacterium Fungus Eutypella sp. BCC 13199. Chemical and tuberculosis: Heterologous Expression, Purification, Pharmaceutical Bulletin, 59(9), 1157-1159. and Enzymatic Characterization of the Encoded 57. Isaka, M., Palasarn, S., Supothina, S., Komwijit, S. and Fructose 1,6-bisphosphatase II. Applied Biochemistry Luangsa-Ard, J.J. (2011). Bioactive Compounds from the and Biotechnology, 164(8), 1376-1389. Scale Insect Pathogenic Fungus Conoideocrella tenuis 47. Haritakun, R., Rachtawee, P., Chanthaket, R., Boonyuen, BCC 18627. Journal of Natural Products, 74(4), 782-789. N. and Isaka, M. (2010). Butyrolactones from the Fungus 58. Isaka, M., Sappan, M., Jennifer Luangsa-ard, J., Hywel- Aspergillus terreus BCC 4651. Chemical and Pharmaceutical Jones, N.L., Mongkolsamrit, S. and Chunhametha, S. Bulletin, 58(11), 1545-1548. (2011). Chemical taxonomy of Torrubiella s. lat.: zeorin 48. Hatin, W.I., Nur-Shafawati, A.R., Zahri, M.K., Xu, S., Jin, L., as a marker of Conoideocrella. Mycological Research, Tan, S.G., Idid, M.R., Zilfalil, B.A. and The HUGO Pan-Asian 115(4-5), 401-405. SNP Consortium. (2011). Population Genetic Structure 59. Isaka, M., Sappan, M., Rachtawee, P. and Boonpratuang, of Peninsular Malaysia Malay Sub-Ethnic Groups. PLoS T. (2011). A tetrahydrobenzofuran derivative from the ONE, 6(4), e18312. fermentation broth of Lentinus squarrosulus BCC 22366. 49. Himananto, O., Thummabenjapone, P., Luxananil, P., Phytochemistry Letters, 4(2), 106-108. Kumpoosiri, M., Hongprayoon, R., Kositratana, W. and 60. Isaka, M., Srisanoh, U., Choowong, W. and Boonpratuang, Gajanandana, O. (2011). Novel and Highly Specific T. (2011). Sterostreins A-E, New Terpenoids from Cultures Monoclonal Antibody to Acidovorax citrulli and of the Basidiomycete Stereum ostrea BCC 22955. Organic Development of ELISA-Based Detection in Cucurbit Letters, doi:10.1021/ol2019778. Leaves and Seed. Plant Disease, 95(9), 1172-1178. 61. Isaka, M., Yangchum, A., Auncharoen, P., Srichomthong, K. 50. Hughes, D.P., Andersen, S.B., Hywel-Jones, N.L., Himaman, and Srikitikulchai, P. (2011). Ring B Aromatic Norpimarane W., Billen, J. and Boomsma, J.J. (2011). Behavioral mecha- Glucoside from a Xylaria sp. Journal of Natural Products, nisms and morphological symptoms of zombie ants 74(2), 300-302. dying from fungal infection. BMC Ecology, 11, art. no. 62. Isaka, M., Yangchum, A., Sappan, M., Suvannakad, R. and 13. Srikitikulchai, P. (2011). Cyclohexadepsipeptides from 51. Imjongjirak, C., Amparyup, P. and Tassanakajon, A. Acremonium sp. BCC 28424. Tetrahedron, doi:10.1016/j. (2011). Two novel antimicrobial peptides, arasin-likeSp tet.2011.08.041. and GRPSp, from the mud crab Scylla paramamosain, 63. Isarankura-Na-Ayudhya, P., Isarankura-Na-Ayudhya, C., exhibit the activity against some crustacean pathogenic Yainoy, Y., Thippakorn, C., Singhagamol, W., Polprachum, bacteria. Fish and Shellfish Immunology, 30(2), 706-712. W., Roytrakul, S. and Prachayasittikul, V. (2010). Proteomic 52. Ingsriswang, S., Yokwai, S. and Wichadakul, D. (2011). alterations of Escherichia coli by paraquat. EXCLI Journal, LinkinPath: From Sequence to Interconnected Pathway. 9(1), 108-118. Bioinformatics, 27(14), 2015-2017. 64. Issarapayup, K., Powtongsook, S. and Pavasant, P. (2011). 53. Intarasirisawat, R., Benjakul, S. and Visessanguan, W. Economical review of Haematococcus pluvialis culture (2011). Chemical compositions of the roes from skipjack, in flat-panel airlift photobioreactors. Aquacultural tongol and bonito. Food Chemistry, 124(4), 1328-1334. Engineering, 44(3), 65-71. 54. Intaraudom, C., Rachtawee, P., Suvannakad, R. and 65. Jantaboon, J., Siangliw, M., Im-mark, S., Jamboonsri, W., Pittayakhajonwut, P. (2011). Antimalarial and antitu- Vanavichit, A. and Toojinda, T. (2011). Ideotype breed- berculosis substances from Streptomyces sp. BCC26924. ing for submergence tolerance and cooking quality by Tetrahedron, 67(39), 7593-7597. marker-assisted selection in rice. Field Crops Research, 123(3), 206-213.
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66. Jindamorakot, S., Am-in, S., Yukphan, P. and Yamada, 77. Kaewsaneha, C., Tangboriboonrat, P., Polpanich, D. and Y. (2011). Limtongia gen. nov. for Zygozyma smithiae Smanmoo, S. (2010). Preparation of core-shell particle of (Lipomycetaceae). Annals of Microbiology, 61, 689-693. disinfectant agent nanocapsules-skim rubber particles 67. Jiravanichpaisal, P., Söderhäll, K. and Söderhäll, I. (2011). by the heterocoagulation technique. Macromolecular Inflammation in Arthropods. Current Pharmaceutical Research, 18(9), 876-883. Design, 16(38), 4166-4174. 78. Kageyama, H., Tripathi, K., Rai, A.K., Cha-Um, S., 68. Jones, E.B.G. (2010). Fungi. Biology Marine Mediteranean, Waditee-Sirisattha, R. and Takabe, T. (2011). An Alkaline 17(1), 681-684. Phosphatase/Phosphodiesterase, PhoD, Induced by Salt 69. Jones, E.B.G. and Pang, K.-L. (2010). 11th International Stress and Secreted Out of the Cells of Aphanothece Marine and Freshwater Mycology Symposium, Taichung, halophytica, a Halotolerant Cyanobacterium. Applied Taiwan R.O.C., November 2009. Botanica Marina, 53(6), and Environmental Microbiology, 77(15), 5178-5183. 475-478. 79. Kanjana, K., Radtanatip, T., Asuvapongpatana, S., 70. Jonmalung, J., Prammananan, T., Leechawengwongs, M. Withyachumnarnkul, B. and Wongprasert, K. (2011). and Chaiprasert, A. (2010). Surveillance of pyrazinamide Solvent extracts of the red seaweed Gracilaria fisheri susceptibility among multidrug-resistant Mycobacterium prevent Vibrio harveyi infections in the black tiger shrimp tuberculosis isolates from Siriraj Hospital, Thailand. BMC Penaeus monodon. Fish and Shellfish Immunology, 30(1), Microbiology, 10, art. no. 223. 389-396. 71. Jupatanakul, N., Wannapapho, W., Eurwilaichitr, L., 80. Kanokratana, P., Uengwetwanit, T., Rattanachomsri, U., Flegel, T.W. and Sritunyalucksana, K. (2011). Cloning Bunterngsook, B., Nimchua, T., Tangphatsornruang, S., and expression of recombinant shrimp PmRab7 (a virus- Plengvidhya, V., Champreda, V. and Eurwilaichitr, L. (2011). binding protein) in Pichia pastoris. Protein Expression and Insights into the Phylogeny and Metabolic Potential Purification, 76(1), 1-6. of a Primary Tropical Peat Swamp Forest Microbial 72. Kaeodee, M., Pongsomboon, S. and Tassanakajon, A. Community by Metagenomic Analysis. Microbial Ecology, (2011). Expression analysis and response of Penaeus 61(3), 518-528. monodon 14-3-3 genes to salinity stress. Comparative 81. Khemkhao, M., Nuntakumjorn, B., Techkarnjanaruk, S. Biochemistry and Physiology - Part B: Biochemistry and and Phalakornkule, C. (2011). Effect of chitosan on UASB Molecular Biology, 159(4), 244-251. treating POME during a transition from mesophilic to 73. Kaewmanee, T., Benjakul, S. and Visessanguan, W. (2011). thermophilic conditions. Bioresource Technology, 102(7), Effect of NaCl on thermal aggregation of egg white 4674-4681. proteins from duck egg. Food Chemistry, 125(2), 706-712. 82. Khongto, B., Laoteng, K. and Tongta, A. (2010). 74. Kaewmanee, T., Benjakul, S. and Visessanguan, W. (2011). Fermentation Process Development of Recombinant Effects of Salting Processes and Time on the Chemical Hansenula polymorpha for Gamma-Linolenic Acid Composition, Textural Properties, and Microstructure Production. Journal of Microbiology and Biotechnology, of Cooked Duck Egg. Journal of Food Science, 76(2), 20(11), 1555-1562. s139-s147. 83. Khongto, B., Laoteng, K. and Tongta, A. (2011). Enhancing 75. Kaewmanee, T., Benjakul, S. and Visessanguan, W. the production of gamma-linolenic acid in Hansenula (2011). Effect of Acetic Acid and Commercial Protease polymorpha by fed-batch fermentation using response Pretreatment on Salting and Characteristics of Salted surface methodology. Chemical Papers, 65(2), 124-131. Duck Egg. Food and Bioprocess Technology, doi:10.1007/ 84. Khonzue, P., Laothanachareon, T., Rattanaphan, N., s11947-011-0510-1. Tinnasulanon, P., Apawasin, S., Paemanee, A., Paemanee, 76. Kaewsaneha, C., Opaprakasit, P., Polpanich, D., V., Ruanglek, V., Tanapongpipat, S., Champreda, V. Smanmoo, S. and Tangboriboonrat, P. (2010). Composite and Eurwilaichitr, L. (2011). Optimization of Xylanase Particles of Disinfectant Nanocapsules-Skim Rubber Production from Aspergillus niger for Biobleaching Latex. International Journal of Polymer Analysis and of Eucalyptus Pulp. Bioscience Biotechnology and Characterization, 15(8), 524-535. Biochemistry, 75(6), 1129-1134.
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85. Kiatmetha, P., Siengdang, W., Bunnag, B., Senapin, S. and 94. Krissanaprasit, A., Somasundrum, M. and Surareungchai, Withyachumnarnkul, B. (2011). Enhancement of survival W. (2011). RGB colour coding of Y-shaped DNA for simul- and metamorphosis rates of Penaeus monodon larvae taneous tri-analyte solid phase hybridization detection. by feeding with the diatom Thalassiosira weissflogii. Biosensors and Bioelectronics, 26(5), 2183-2187. Aquaculture International, doi:10.1007/s10499-010- 95. Kritsiriwuthinan, K., Chaotheing, S., Shaw,P.J., 9375-y. Wongsombat, C., Chavalitshewinkoon-Petmitr, P. and 86. Kitidee, K., Nangola, S., Gonzalez, G., Boulanger, P., Kamchonwongpaisan, S. (2011). Global gene expression Tayapiwatana, C. and Hong, S.S. (2010). Baculovirus profiling of Plasmodium falciparum in response to the display of single chain antibody (scFv) using a novel anti-malarial drug pyronaridine. Malaria Journal, 10, 242. signal peptide. BMC Biotechnology, 10, 80. 96. Kümpornsin, K., Jiemsup, S., Yongkiettrakul, S. and 87. Klinbunga, S., Yuvanatemiya, V., Wongphayak, S., Khetpu, Chookajorn, T. (2011). Characterization of band 3–an- K., Menasveta, P. and Khamnamtong, B. (2010). Genetic kyrin–Protein 4.2 complex by biochemical and mass population differentiation of the blue swimming crab spectrometry approaches. Biochemical and Biophysical Portunus pelagicus (Portunidae) in Thai waters revealed Research Communications, 406(3), 332-335. by RAPD analysis. Genetics and Molecular Research, 9(3), 97. Kunanopparat, A., Chaivisuthangkura, P., Senapin, 1615-1624. S., Longyant, S., Rukpratanporn, S. Flegel, T.W. and 88. Kommanee, J., Tanasupawat, S., Yukphan, P., Malimas, Sithigorngul, P. (2011). Detection of infectious myo- T., Muramatsu, Y., Nakagawa, Y. and Yamada Y. (2011). necrosis virus using monoclonal antibody specific to Gluconobacter nephelii sp. nov., an acetic acid bacterium N and C fragments of the capsid protein expressed in the α-Proteobacteria. International Journal of Systematic heterologously. Journal of Virological Methods, 171(1), and Evolutionary Microbiology, PMID: 20870887. 141-148. 89. Koowadjanakul, N., Jindamorakot, S., Yongmanitchai, 98. Kurdrid, P., Senachak, J., Sirijuntarut, M., Yutthanasirikul, W. and Limtong, S. (2011). Ogataea phyllophila sp. nov., R., Phuengcharoen, P., Jeamton, W., Roytrakul, S., Candida chumphonensis sp. nov. and Candida mat- Cheevadhanarak, S. and Hongsthong, A. (2011). tranensis sp. nov., three methylotrophic yeast species Comparative analysis of the Spirulina platensis subcellular from phylloplane in Thailand. Antonie van Leeuwenhoek proteome in response to low- and high-temperature Journal of Microbiology, 100(2), 207-217. stresses: uncovering cross-talk of signaling components. 90. Korinsak, S., Sirithunya, P., Meakwatanakarn, P., Sarkarung, Proteome Science, 9, 39. S., Vanavichit, A. and Toojinda, T. (2011). Changing allele 99. Laoteng, K., Čertík, M. and Cheevadhanark, S. (2011). frequencies associated with specific resistance genes Mechanisms controlling lipid accumulation and poly- to leaf blast in backcross introgression lines of Khao unsaturated fatty acid synthesis in oleaginous fungi. Dawk Mali 105 developed from a conventional selec- Chemical Papers, 65(2), 97-103. tion program. Field Crops Research, 122(1), 32-39. 100. Leelatanawit, R., Uawisetwathana, U., Klinbunga, S. 91. Kornsakulkarn, J., Dolsophon, K., Boonyuen, N., and Karoonuthaisiri, N. (2011). A cDNA microarray, Boonruangprapa, T., Rachtawee, P., Prabpai, S., UniShrimpChip, for identification of genes relevant Kongsaeree, P. and Thongpanchang, C. (2011). to testicular development in the black tiger shrimp Dihydronaphthalenones from endophytic fungus (Penaeus monodon). BMC Molecular Biology, 12, 15. Fusarium sp. BCC 14842. Tetrahedron, 67(39), 7540-7547. 101. Limpiti, T., Intarapanich, A., Assawamakin, A., Shaw, 92. Kositcharoenkul, N., Chatchawankanphanich, O., P.J., Wangkumhang, P., Piriyapongsa, J., Ngamphiw, Bhunchoth, A. and Kositratana, W. (2011). Detection of C. and Tongsima, S. (2011). Study of large and highly Xanthomonas citri subsp. citri from field samples using stratified population datasets by combining iterative single-tube nested PCR. Plant Pathology, 60(3), 436-442. pruning principal component analysis and Structure. 93. Krajaejun, T., Khositnithikul, R., Lerksuthirat, T., Lowhnoo, BMC Bioinformatics, 12, 255. T., Rujirawat, T., Petchthong, T., Yingyong, W., Suriyaphol, 102. Limtong, S., Jindamorakot, S., Am-In, S., Kaewwichian, P., Smittipat, N., Juthayothin, T., Phuntumart, V. and R., Nitiyon, S., Yongmanitchai, W. and Nakase, T. (2011). Sullivang, T.D. (2011). Expressed sequence tags reveal Candida uthaithanina sp. nov., an anamorphic yeast genetic diversity and putative virulence factors of the species in Nakaseomyces clade isolated in Thailand. pathogenic oomycete Pythium insidiosum. Fungal Biology, Antonie van Leeuwenhoek Journal of Microbiology, 99(4), 115(7), 683-696. 865-871.
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103. Liu J.-K., Chomnunti P., Cai L., Phookamsak R., Chukeatirote 112. Montet, D., Alldrick, A., Bordier, M., Bresson, H., E., Jones E.B.G., Moslem M. and Hyde K.D. (2010). Chokesajjawatee, N., Durand, N., Toan Ha, T., Chea Hak, Phylogeny and morphology of Neodeightonia palmicola S., Hariyadi, P., Jinap, S., Keeratipibul, S., Leepipatpiboon, sp. nov. from palms. Sydowia, 62(2), 261-276. N., Quang Luong, H., Marvin, H.J.P., Medoc, J.M., Moustier, 104. Longyant, S., Chaiyasittrakul, K., Rukpratanporn, S., P., Nitisinprasert, S., Dong Phan, T., Poms, R., Prasertvit, Chaivisuthangkura, P. and Sithigorngul, P. (2011). Simple S., Rakshit, S.K., Ruangwises, S., Shamsudin, M.N., and direct detection of Aeromonas hydrophila infection Siriwatwechakul, W., Sparringa, R.A., Stouten, P., Stroka, in the goldfish, Carassius auratus (L.), by dot blotting J., Taharnklaew, R., Tayaputch, N., Tongpim, S., Valyasevi, using specific monoclonal antibodies. Journal of Fish R., Vithayarungruangsri, J. and Saletes, S. (2010). Future Diseases, doi:10.1111/j.1365-2761.2010.01197.x. topics of common interest for EU and SEA partners in 105. Luangsa-ard, J., Houbraken, J., Van Doorn, T., Hong, S.- food quality, safety and traceability. Quality Assurance B., Borman, A.M., Hywel-Jones, N.L. and Samson, R.A. and Safety of Crops and Foods, 2(4), 158-164. (2011). Purpureocillium, a new genus for the medically 113. Morchang, A., Yasamut, U., Netsawang, J., Noisakran, S., important Paecilomyces lilacinus. FEMS Microbiology Wongwiwat, W., Songprakhon, P., Srisawat, C., Puttikhunt, Letters, 321(2), 141-149. C., Kasinrerk, W., Malasit, P., Yenchitsomanus, P. and 106. Luangsa-ard, J.J., Ridkaew, R., Tasanathai, K., Limjindaporn, T. (2011). Cell death gene expression Thanakitpipattana, D. and Hywel-Jones, N. (2011). profile: Role of RIPK2 in dengue virus-mediated apop- Ophiocordyceps halabalaensis: a new species of tosis. Virus Research, 156(1-2), 25-34. Ophiocordyceps pathogenic to Camponotus gigas in Hala 114. Muangkeow, B., Ikejima, K., Powtongsook, S. and Gallardo, Bala Wildlife Sanctuary, Southern Thailand. Mycological W. (2011). Growth and nutrient conversion of white Research, 115(7), 608-614. shrimp Litopenaeus vannamei (Boone) and Nile tilapia 107. Mcconkey, K.R. and Brockelman, W.Y. (2011). Oreochromis niloticus L. in an integrated closed recircu- Nonredundancy in the dispersal network of a generalist lating system. Aquaculture Research, 42(9), 1246-1260. tropical forest tree. Ecology, 92(7), 1492-1502. 115. Nagila, A., Netsawang, J., Srisawat, C., Noisakran, S., 108. Methacanon, P., Weerawatsophon, U., Tanjak, P., Morchang, A., Yasamut, U., Puttikhunt, C., Kasinrerk, W., Rachtawee, P. and Prathumpai, W. (2011). Interleukin-8 Malasit, P., Yenchitsomanus, P.T. and Limjindaporn, T. stimulating activity of low molecular weight β-glucan (2011). Role of CD137 signaling in dengue virus-medi- depolymerized by γ-irradiation. Carbohydrate Polymers, ated apoptosis. Biochemical and Biophysical Research 86(2), 574-580. Communications, 410(3), 428-433. 109. Midgley, C.M., Bajwa-Joseph, M., Vasanawathana, S., 116. Nakase, T., Jindamorakot, S., Am-In, S., Lee, C.F. and Limpitikul, W., Wills, B., Flanagan, A., Waiyaiya, E., Tran, Limtong, S. (2011). Four novel species of the anamorphic H.B., Cowper, A.E., Chotiyarnwon, P., Grimes, J.M., Yoksan, yeast genus Candida found in Thailand and Taiwan. S., Malasit, P., Simmons, C.P., Mongkolsapaya, J. and Journal of General and Applied Microbiology, 57(4), 231- Screaton, G.R. (2011). An In-Depth Analysis of Original 242. Antigenic Sin in Dengue Virus Infection. Journal of 117. Nakase, T., Jindamorakot, S., Am-In, S., Lee, C.-F., Imanishi, Virology, 85(1), 410-421. Y. and Limtong, S. (2011). Three novel species of the ana- 110. Mitprasat, M., Roytrakul, S., Jiemsup, S., Boonseng, O. and morphic yeast genus Candida in the Candida intermedia Yokthongwattana, K. (2011). Leaf proteomic analysis in clade found in Japan, Thailand and Taiwan. Journal of cassava (Manihot esculenta, Crantz) during plant devel- General and Applied Microbiology, 57(2), 73-81. opment, from planting of stem cutting to storage root 118. Nakase, T., Jindamorakot, S., Am-In, S., Ninomiya, S., formation. Planta, 233(6), 1209-1221. Kawasaki, H. and Limtong, S. (2011). Candida maleeae 111. Mongkolsamrit, S., Luangsa-ard, J.J. and Hywel-Jones, N.L. sp. nov., a novel anamorphic yeast species in the (2011). Samuelsia mundiveteris sp. nov. from Thailand. Ambrosiozyma clade found in Thailand. Journal of Mycologia, 103(4), 921-927. General and Applied Microbiology, 57(4), 253-258.
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119. Nakase, T., Jindamorakot, S., Imanishi, Y., Am-In, S., 129. Palangsuntikul, R., Somasundrum, M. and Surareungchai, Ninomiya, S., Kawasaki, H. and Limtong, S. (2010). W. (2010). Kinetic and analytical comparison of horserad- Candida potacharoeniae sp. nov. and Candida spenceri ish peroxidase on bare- and redox-modified single-walled sp. nov., two novel galactose-containing ascomycetous carbon nanotubes. Electrochimica Acta, 56(1), 470-475. anamorphic yeast species isolated in Thailand. Journal 130. Pang, K.-L., Alias, S.A., Chiang, M.W.L., Vrijmoed, L.L.P. of General and Applied Microbiology, 56(4), 287-295. and Jones, E.B.G. (2010). Sedecimiella taiwanensis gen. et 120. Nerungsi, C., Wanitchang, P., Sahasithiwat, S., Sadorn, K., sp. nov., a marine mangrove fungus in the Hypocreales Kerdcharoen, T. and Thongpanchang, T. (2010). Organic (Hypocreomycetidae, Ascomycota). Botanica Marina, electroluminescence devices based on anthracene sul- 53(6), 493-498. fide derivatives.Tetrahedron Letters, 51(49), 6392-6395. 131. Panphut, W., Senapin, S., Sriurairatana, S., 121. Ngamphiw, C., Assawamakin, A., Xu, S., Shaw, P.J., Yang, Withyachumnarnkul, B. and Flegel, T.W. (2011). A novel J.O., Yang, J.O., Ghang, H., Bhak, J., Liu, E., Tongsima, integrase-containing element may interact with Laem- S. and the HUGO Pan-Asian SNP Consortium. (2011). Singh virus (LSNV) to cause slow growth in giant tiger PanSNPdb: The Pan-Asian SNP Genotyping Database. shrimp. BMC Veterinary Research, 7, 18. PLoS ONE, 6(6), e21451. 132. Pengsuk, C., Longyant, S., Rukpratanporn, S., 122. Nicholson, C.O., Costin, J.M., Rowe, D.K., Lin, L., Chaivisuthangkura, P., Sridulyakul, P. and Sithigorngul Jenwitheesuk, E., Samudrala, R., Isern, S. and Michael, P. (2011). Differentiation among the Vibrio cholerae S.F. (2011). Viral entry inhibitors block dengue antibody- serotypes O1, O139, O141 and non-O1, non-O139, non- dependent enhancement in vitro. Antiviral Research, O141 using specific monoclonal antibodies with dot 89(1), 71-74. blotting. Journal of Microbiological Methods, doi:10.1016/j. 123. Nishimura, T., Cha-Um, S., Takagaki, M., Ohyama, K. and mimet.2011.07.022. Kirdmanee, C. (2011). Survival percentage, photosyn- 133. Pichyangkul, S., Jongkaewwattana, A., Thitithanyanont, thetic abilities and growth characters of two indica rice A., Ekchariyawat, P., Wiboon-ut, S., Limsalakpetch, (Oryza sativa L. spp. indica) cultivars in response to iso A., Yongvanitchit, K., Kum-Arb, U., Mahanonda, R., osmotic stress. Spanish Journal of Agricultural Research, Utaisincharoen, P., Sirisinha, S., Mason, C.J. and Fukuda, 9(1), 262-270. M.M. (2010). Cross-reactive antibodies against avian 124. Nivitchanyong, T., Yongkiettrakul, S., Kramyu, J., influenza virus A (H5N1). Emerging Infectious Diseases, Pannengpetch, S. and Wanasen, N. (2011). Enhanced 15(9), 1537-1539. expression of secretable influenza virus neuraminidase 134. Pimpha, N., Chaleawlert-umpon, S., Chruewkamlow, in suspension mammalian cells by influenza virus N. and Kasinrerk, W. (2011). Preparation of anti-CD4 nonstructural protein 1. Journal of Virological Methods, monoclonal antibody-conjugated magnetic poly(glycidyl doi:10.1016/j.jviromet.2011.08.010. methacrylate) particles and their application on CD4+ 125. Noonin, C., Jiravanichpaisal, P., Söderhäll, I., Merino, S., lymphocyte separation. Talanta, 84(1), 89-97. Tomás, J.M. and Söderhäll, K. (2010). Melanization and 135. Pinijsuwan, S., Rijiravanich, P., Somasundrum, M. and Pathogenicity in the Insect, Tenebrio molitor, and the Surareungchai, W. (2010). Attomolar Electrochemical Crustacean, Pacifastacus leniusculus, by Aeromonas Detection of DNA Hybridization Based on Enhanced hydrophila AH-3. PLoS ONE, 5(12), art. no. e15728. Latex/Gold Nanoparticles. Advanced Engineering Materials, 126. Nootong, K., Pavasant, P. and Powtongsook, S. (2011). 12(11), B649-B653. Effects of Organic Carbon Addition in Controlling 136. Pitakdantham, W., Sutabutra, T., Chiemsombat, P. and Inorganic Nitrogen Concentrations in a Biofloc System. Pitaksutheepong, C. (2010). Isolation and characterization Journal of the World Aquaculture Society, 42(3), 339-346. of chalcone synthase gene isolated from Dendrobium 127. Nuangnaowarat, W., Phupong, W. and Isaka, M. (2010). Sonia earsakul. Pakistan Journal of Botany, 13(20), 1000- New xanthones from the barks of Cratoxylum sumatra- 1005. num ssp. neriifolium. Heterocycles, 81(10), 2335-2341. 137. Pitakdantham, W., Sutabutra, T., Chiemsombat, P. and 128. Nuangnaowarat, W., Phupong, W., Intereya, K. and Isaka, Pitaksutheepong, C. (2011). Isolation and Characterization M. (2010). New prenylated xanthone from the branch of Dihydroflavonol 4-reductase Gene in Dendrobium of Garcinia costata. Heterocycles, 81(10), 2377-2384. Flowers. Journal of Plant Sciences, 6(2), 88-94.
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138. Plevka, P., Battisti, A.J., Junjhon, J., Winkler, D.C., Holdaway, 146. Pulmanausahakul, R., Roytrakul, S., Auewarakul, P. and H.A., Keelapang, P., Sittisombut, N., Kuhn, R.J., Steven, A.C. Smith, D.R. (2011). Chikungunya in Southeast Asia: and Rossmann, M.G. (2011). Maturation of flaviviruses understanding the emergence and finding solutions. starts from one or more icosahedrally independent International Journal of Infectious Diseases, doi:10.1016/j. nucleation centres. EMBO reports, 12(6), 602-606. ijid.2011.06.002. 139. Pongsomboon, S., Tang, S., Boonda, S., Aoki, T., Hirono, 147. Punjaisee, C., Chaiyasut, C., Chansakaow, S., Tharata, I. and Tassanakajon, A. (2011). A cDNA microarray ap- S., Visessanguan, W. and Punjaisee, S. (2011). proach for analyzing transcriptional changes in Penaeus 8-Hydroxygenistein formation of soybean fermented monodon after infection by pathogens. Fish and Shellfish with Aspergillus oryzae BCC 3088. African Journal of Immunology, 30(1), 439-446. Agricultural Research, 6(4), 785-789. 140. Ponnikorn, S., Panichakul, T., Sresanga, K., Wongborisuth, 148. Puttharugsa, C., Wangkam, T., Huangkamhang, N., C., Roytrakul, S., Hongeng, S. and Tungpradabkul, S. (2011). Gajanandana, O., Himananto, O., Sutapun, B., Amarit, R., Phosphoproteomic analysis of apoptotic hematopoietic Somboonkaew, A. and Srikhirin, T. (2011). Development stem cells from hemoglobin E/beta-thalassemia. Journal of surface plasmon resonance imaging for detection of Translational Medicine, 9, 96. of Acidovorax avenae subsp. citrulli (Aac) using specific 141. Ponza, P., Yocawibun, P., Sittikankaew, K., Hiransuchalert, monoclonal antibody. Biosensors and Bioelectronics, R., Yamano, K. and Klinbunga, S. (2011). Molecular clon- 26(5), 2341-2346. ing and expression analysis of the Mitogen-activating 149. Puttikhunt, C., Prommool, T., U-thainual, N., Ong- protein kinase 1 (MAPK1) gene and protein during ovarian ajchaowlerd, P., Yoosook, K., Tawilert, C., Duangchinda, development of the giant tiger shrimp Penaeus mon- T., Jairangsri, A., Tangthawornchaikul, N., Malasit, P. odon. Molecular Reproduction and Development, 78(5), and Kasinrerk, W. (2011). The development of a novel 347-360. serotyping-NS1-ELISA to identify serotypes of dengue 142. Prammananan, T., Chaiprasert, A. and Leechawengwongs, virus. Journal of Clinical Virology, 50(4), 314-319. M. (2011). 8-years experience of fluoroquinolone sus- 150. Rachapradit, N., Thepa, S. and Monyakul, V. (2011). An ceptibility testing of multidrug-resistant Mycobacterium Influence of Air Volume Flow Rate and Temperature tuberculosis isolates from Siriraj Hospital, Thailand. Set Point on Performance of Inverter Split-Type Air- International Journal of Antimicrobial Agents, 37(1), 84-85. Conditioner. Experimental Techniques, doi:10.1111/j.1747- 143. Prapavorarat, A., Pongsomboon, S.and Tassanakajon, A. 1567.2011.00720.x. (2010). Identification of genes expressed in response 151. Raita, M., Laothanachareon, T., Champreda, V. and to yellow head virus infection in the black tiger shrimp, Laosiripojana, N. (2011). Biocatalytic esterification of Penaeus monodon, by suppression subtractive hybrid- palm oil fatty acids for biodiesel production using ization. Developmental and Comparative Immunology, glycine-based cross-linked protein coated microcrys- 34(6), 611-617. talline lipase. Journal of Molecular Catalysis B-Enzymatic, 144. Preechaphol, R., Klinbunga, S., Khamnamtong, B. and doi:10.1016/j.molcatb.2011.07.020. Menasveta, P. (2010). Isolation and characterization of 152. Rattanachomsri, U., Kanokratana, P., Eurwilaichitr, genes functionally involved in ovarian development of L., Igarashi, Y. and Champreda, V. (2011). Culture- the giant tiger shrimp Penaeus monodon by suppression independent phylogenetic analysis of the microbial subtractive hybridization (SSH). Genetics and Molecular community in industrial sugarcane bagasse feedstock Biology, 33(4), 676-685. piles. Bioscience Biotechnology and Biochemistry, 75(2), 145. Prompamorn, P., Sithigorngul, P., Rukpratanporn, S., 232-239. Longyant, S., Sridulyakul, P. and Chaivisuthangkura, P. 153. Rehner, S.A., Minnis, D., Sung, G.H., Luangsa-ard, J.J., (2011). The development of loop-mediated isothermal DeVotto, L. and Humber, R.A. (2011). Phylogeny and amplification combined with lateral flow dipstick for systematics of the anamorphic, entomopathogenic detection of Vibrio parahaemolyticus. Letters in Applied genus Beauveria. Mycologia, doi:10.3852/10-302. Microbiology, doi:10.1111/j.1472-765X.2011.03007.x. 154. Ritthong, C., Puanglarp, N., Piyatiratitivorakul, S. and Menasvata, P. (2011). Biomonitoring of Mercury Contamination at Petroleum Production Platforms in the Gulf of Thailand using Transplanted Green Mussel, Perna viridis. Environment Asia, 4(1), 21-29.
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155. Rodaree, K., Maturos, T., Chaotheing, S., Pogfay, T., 163. Sappat, A., Jaroenram, W., Puthawibool, T., Lomas, T., Suwanakitti, N., Wongsombat, C., Jaruwongrungsee, K., Tuantranont, A. and Kiatpathomchai, W. (2011). Detection Wisitsoraat, A., Kamchonwongpaisan, S., Lomas, T. and of shrimp Taura syndrome virus by loop-mediated iso- Tuantranont, A. (2011). DNA hybridization enhancement thermal amplification using a designed portable multi- using piezoelectric microagitation through a liquid channel turbidimeter. Journal of Virological Methods, coupling medium. Lab on a Chip, 11(6), 1059-1064. 175(2), 141-148. 156. Roobsoong, W., Roytrakul, S., Sattabongkot, J., Li, J., 164. Saricheewin, K., Sirivithayapakorn, S., Noophan, P., Udomsangpetch, R. and Cui, L. (2011). Determination of Wantawin, C., Techkarnjanarunk, S. and Munakata-Marr, the Plasmodium vivax schizont stage proteome. Journal J. (2010). Nitrogen removal of anammox cultures under of Proteomics, 74(9), 1701-1710. different enrichment conditions.Journal of Environmental 157. Roongsawang, N., Washio, K. and Morikawa, M. (2011). Science and Health, Part A, 45(14), 1832-1838. Diversity of Nonribosomal Peptide Synthetases Involved 165. Sawasdee, N., Junking, M., Ngaojanlar, P., Sukomon, N., in the Biosynthesis of Lipopeptide Biosurfactants. Ungsupravate, D., Limjindaporn, T., Akkarapatumwong, International Journal of Molecular Sciences, 12(1), 141-172. V., Noisakran, S. and Yenchitsomanus, P.-T. (2010). Human 158. Ruenwai, R., Neiss, A., Laoteng, K., Vongsangnak, W., kidney anion exchanger 1 interacts with adaptor-related Dalfard, A.B., Cheevadhanarak, S., Petranovic, D. and protein complex 1 μ1A (AP-1 mu1A). Biochemical and Nielsen, J. (2011). Heterologous production of polyun- Biophysical Research Communications, 401(1), 85-91. saturated fatty acids in Saccharomyces cerevisiae causes 166. Searmsirimongkol, P., Rangsunvigit, P., Leethochawalit, a global transcriptional response resulting in reduced M. and Chavadeja, S. (2011). Hydrogen production from proteasomal activity and increased oxidative stress. alcohol distillery wastewater containing high potas- Biotechnology Journal, 6(3), 343-356. sium and sulfate using an anaerobic sequencing batch 159. Rungnoi, O., Chanprem, S., Toojinda, T., Godwin, I., reactor. International Journal of Hydrogen Energy, 36(20), Lambrides, C. and Srinives, P. (2010). Characterization, 12810-12821. inheritance, and molecular study of opaque leaf mutant 167. Seepiban, C., Gajanandana, O., Attathom, T. and Attathom, in mungbean (Vigna radiata (L.) Wilczek). Journal of Crop S. (2011). Tomato necrotic ringspot virus, a new tospo- Science and Biotechnology, 13(4), 219-226. virus isolated in Thailand. Archives of Virology, 156(2), 160. Rungroj, N., Sritippayawan, S., Thongnoppakhun, W., 263-274. Paemanee, A., Sawasdee, N., Nettuwakul, C., Sudtachat, 168. Senapin, S., Phiwsaiya, K., Gangnonngiw, W. and Flegel, N., Ungsupravate, D., Praihirunkit, P., Chuawattana, D., T.W. (2011). False rumours of disease outbreaks caused Akkarapatumwong, V., Borvornpadungkitti, S., Susaengrat, by infectious myonecrosis virus (IMNV) in the whiteleg W., Vasuvattakul, S., Malasit, P. and Yenchitsomanus, P. shrimp in Asia. Journal of Negative Results in BioMedicine, (2011). Prothrombin Haplotype Associated With Kidney 10, 10. Stone Disease in Northeastern Thai Patients. Urology, 169. Senapin, S., Phiwsaiya, K., Kiatmetha, P. and 77(1), 249.e17-249.e23. Withyachumnarnkul, B. (2011). Development of primers 161. Sakayaroj, J., Pang, K.-L. and Jones, E.B.G. (2011). Multi- and a procedure for specific identification of the diatom gene phylogeny of the Halosphaeriaceae: its ordinal Thalassiosira weissflogii. Aquaculture International, 19(4), status, relationships between genera and morphological 693-704. character evolution. Fungal Diversity, 46(1), 87-109. 170. Shoombuatong, W., Traisathit, P., Prasitwattanaseree, 162. Saksmerprome, V., Jitrakorn, S., Chayaburakul, K., S., Tayapiwatana, C., Cutler, R. and Chaijaruwanich, J. Laiphrom, S., Boonsua, K. and Flegel, T.W. (2011). (2011). Prediction of the disulphide bonding state of Additional random, single to multiple genome frag- cysteines in proteins using Conditional Random Fields. ments of Penaeus stylirostris densovirus in the giant International Journal of Data Mining and Bioinformatics, tiger shrimp genome have implications for viral disease 5(4), 449-464. diagnosis. Virus Research, 160, 180-190. 171. Siringam, K., Juntawong, N., Cha-um, S. and Kirdmanee, C. (2011). Salt stress induced ion accumulation, ion homeostasis, membrane injury and sugar contents in salt-sensitive rice (Oryza sativa L. spp. indica) roots under isoosmotic conditions. African Journal of Biotechnology, 10(8), 1340-1346.
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172. Sithigorngul, P., Rukpratanporn, S., Chaivisuthangkura, 181. Sriket, C., Benjakul, S., Visessanguan, W. and Hara, K. P., Sridulyakul, P. and Longyant, S. (2011). Simultaneous (2011). Effect of legume seed extracts on the inhibi- and rapid detection of white spot syndrome virus and tion of proteolytic activity and muscle degradation of yellow head virus infection in shrimp with a dual im- fresh water prawn (Macrobrachium rosenbergii). Food munochromatographic strip test. Journal of Virological Chemistry, 129(3), 1093-1099. Methods, 173(1), 85-91. 182. Sriket, C., Benjakul, S., Visessanguan, W. and Kishimura, 173. Smanmoo, S., Nasomphan, W. and Tangboriboonrat, H. (2011). Collagenolytic serine protease in fresh water P. (2011). Highly selective fluorescent chemosensor prawn (Macrobrachium rosenbergii): Characteristics and for Fe3+ imaging in living cells. Inorganic Chemistry its impact on muscle during iced storage. Food Chemistry, Communications, 14(2), 351-354. 124(1), 29-35. 174. Smanmoo, S., Nasomphan, W. and Tangboriboonrat, P. 183. Sritakae, A., Praseartkul, P., Cheunban, W., Miphokasap, (2011). Isothiocyanatoluminol as a Chemiluminescence P., Eiumnoh, A., Burns, P., Phironrit, N., Phuangrat, B., Labeling Reagent for Amino Acids and Proteins. Chemistry Kitsubun, P. and Meechai, A. (2011). Mapping airborne Letters, 40(2), 188-190. pollen of papaya (Carica papaya L.) and its distribution 175. Somboonna, N., Mangkalanan, S., Udompetcharaporn, related to land use using GIS and remote sensing. A., Krittanai, C., Sritunyalucksana, K. and Flegel, T.W. Aerobiologia, doi:10.1007/s10453-011-9197-z. (2010). Mud crab susceptibility to disease from white 184. Sritunyalucksana, K., Intaraprasong, A., Sa-Nguanrut, spot syndrome virus is species-dependent. BMC Research P., Filer, K. and Fegan, D.F. (2011). Organic selenium Notes, 3, art. no. 315. supplementation promotes shrimp growth and disease 176. Somsak, V., Srichairatanakool, S., Kamchonwongpaisan, resistance to Taura syndrome virus. ScienceAsia, 37(1), S., Yuthavong, Y. and Uthaipibull, C. (2011). Small-scale 24-30. in vitro culture and purification of Plasmodium berghei 185. Sumpavapol, P., Tongyonk, L., Tanasupawat, S., for transfection experiment. Molecular and Biochemical Chokesajjawatee, N., Luxananil, P. and Visessanguan, W. Parasitology, 177(2), 156-159. (2010). Bacillus siamensis sp. nov., isolated from salted 177. Songsiriritthigul, C., Lapboonrueng, S., Roytrakul, S., crab (poo-khem) in Thailand. International Journal of Haltrich, D. and Yamabhai, M. (2011). Crystallization and Systematic and Evolutionary Microbiology, 60(10), 2364- preliminary crystallographic analysis of β-mannanase 2370. from Bacillus licheniformis. Acta Crystallographica Section 186. Sunthornvarabhas, J., Chatakanonda, P., Piyachomkwan, F: Structural Biology and Crystallization Communications, K. and Sriroth, K. (2011). Electrospun polylactic acid and 67(2), 217-220. cassava starch fiber by conjugated solvent technique. 178. Soowannayan, C. and Phanthura, M. (2011). Horizontal Materials Letters, 65(6), 985-987. transmission of white spot syndrome virus (WSSV) be- 187. Supaibulwattana, K., Kuntawunginn, W., Cha-um, S. tween red claw crayfish (Cherax quadricarinatus) and and Kirdmanee, C. (2011). Artemisinin accumulation the giant tiger shrimp (Penaeus monodon). Aquaculture, and enhanced net photosynthetic rate in Qinghao
319, 5-10. (Artemisia annua L.) hardened in vitro in enriched-CO2 179. Sraphet, S., Boonchanawiwat, A., Thanyasiriwat, T., photoautotrophic conditions. Plant Omics, 4(2), 75-81. Boonseng, O., Tabata, S., Sasamoto, S., Shirasawa, K., 188. Surasilp, T., Longyant, S., Rukpratanporn, S., Sridulyakul, Isobe, S., Lightfoot, D.A., Tangphatsornruang, S. and P., Sithigorngul, P. and Chaivisuthangkura, P. (2011). Triwitayakorn, K. (2011). SSR and EST-SSR-based genetic Rapid and sensitive detection of Vibrio vulnificus by linkage map of cassava (Manihot esculenta Crantz). loop-mediated isothermal amplification combined with Theoretical and Applied Genetics, 122(6), 1161-1170. lateral flow dipstick targeted to rpoS gene. Molecular 180. Sriket, C., Benjakul, S. and Visessanguan, W. (2011). and Cellular Probes, 25(4), 158-163. Characterisation of proteolytic enzymes from muscle and 189. Suthianthong, P., Pulsook, N., Supungul, P., Tassanakajon, hepatopancreas of fresh water prawn (Macrobrachium A. and Rimphanitchayakit, V. (2011). A double WAP rosenbergii). Journal of the Science of Food and Agriculture, domain-containing protein PmDWD from the black 91(1), 52-59. tiger shrimp Penaeus monodon is involved in the con- trolling of proteinase activities in lymphoid organ. Fish and Shellfish Immunology, 30(3), 783-790.
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190. Tanasupawat, S., Kommanee, J., Yukphan, P., Nakagawa, 199. Tran, T., Lee, B.H., Yang, H.S., Chotineeranat, S., Sriroth, K. Y. and Yamada, Y. (2011). Identification of Acetobacter and Kim, H.J. (2011). Use of starch granules melting to strains from Thai fermented rice products based on the control the properties of bio-flour filled polypropylene 16S rRNA gene sequence and 16S-23S rRNA gene internal and poly(butylene succinate) composites:Mechanical transcribed spacer restriction analyses. Journal of the properties. Starch- Stärke, doi:10.1002/star.201100024. Science of Food and Agriculture, doi: 10.1002/jsfa.4506. 200. Trisuwan, K., Khamthong, N., Rukachaisirikul, V., 191. Tanasupawat, S., Kommanee, J., Yukphan, P., Phongpaichit, S., Preedanon, S. and Sakayaroj, J. (2010). Muramatsu, Y., Nakagawa, Y. and Yamada, Y. (2011). Anthraquinone, Cyclopentanone, and Naphthoquinone Acetobacter farinalis sp. nov., an acetic acid bacterium Derivatives from the Sea Fan-Derived Fungi Fusarium in the α-Proteobacteria. Journal of General and Applied spp. PSU-F14 and PSU-F135. Journal of Natural Products, Microbiology, 57(3), 159-167. 73(9), 1507-1511. 192. Tangphatsornruang, S., Uthaipaisanwong, P., 201. Trisuwan, K., Rukachaisirikul, V., Kaewpet, M., Phongpaichit, Sangsrakru, D., Chanprasert, J., Yoocha, T., Jomchai, N. S., Hutadilok-Towatana, N., Preedanon, S. and Sakayaroj, and Tragoonrung, S. (2011). Characterization of the J. (2011). Sesquiterpene and Xanthone Derivatives from complete chloroplast genome of Hevea brasiliensis the Sea Fan-Derived Fungus Aspergillus sydowii PSU-F154. reveals genome rearrangement, RNA editing sites and Journal of Natural Products, 74(7), 1663-1667. phylogenetic relationships. Gene, 475(2), 104-112. 202. Trisuwan, K., Rukachaisirikul, V., Phongpaichit, S., 193. Tangsongcharoen, C., Boonserm, P. and Promdonkoy, Preedanon, S. and Sakayaroj, J. (2011). Modiolide and B. (2011). Functional characterization of truncated frag- pyrone derivatives from the sea fan-derived fungus ments of Bacillus sphaericus binary toxin BinB. Journal Curvularia sp. PSU-F22. Archives of Pharmacal Research, of Invertebrate Pathology, 106(2), 230-235. 34(5), 709-714. 194. Techaoei, S., Lumyong, S., Prathumpai, W., Santiarwarn, D. 203. Tsaia, J.J., Jen, Y.H., Chang, J.S., Hsiao, H.M., Noisakran, and Leelapornpisid, P. (2011). Screening Characterization S. and Perng, G.C. (2011). Frequency Alterations in Key and Stability of Biosurfactant produced by Pseudomonas Innate Immune Cell Components in the Peripheral aeruginosa SCMU106 Isolated from Soil in Northern Blood of Dengue Patients Detected by FACS Analysis. Thailand. Asian Journal of Biological Sciences, 4(4), 340- Journal of Innate Immunity, 3(5), 530-540. 351. 204. Ubol, S., Suksatu, A., Modhiran, N., Sangma, C., 195. Thummajitsakul, S., Klinbunga, S. and Sittipraneed, S. Thitithanyanont, A., Fukuda, M. and Juthayothin, T. (2011). (2011). Genetic Differentiation of the Stingless Bee Intra-host diversities of receptor-binding domain of stork Tetragonula pagdeni in Thailand Using SSCP Analysis faeces-derived avian H5N1 viruses and its significance of a Large Subunit of Mitochondrial Ribosomal DNA. as predicted by molecular dynamic simulation. Journal Biochemical Genetics, 49(7-8), 499-510. of General Virology, 92(2), 307-314. 196. Tipsuwan, W., Srichairatanakool, S., Kamchonwongpaisan, 205. Uppanan, P., Channasanon, S., Veeranondh, S. and S.,Yuthavong, Y. and Uthaipibull, C. (2011). Selection Tanodekaew, S. (2011). Synthesis of GTMAC modified of drug resistant mutants from random library of chitin-PAA gel and evaluation of its biological properties. Plasmodium falciparum dihydrofolate reductase in Journal of Biomedical Materials Research Part A, 98A(2), Plasmodium berghei model. Malaria Journal, 10, 119. 185-191. 197. Tosukhowong, A., Visessanguan, W., Pumpuang, L., 206. Vanavichit, A. and Yoshihashi, T. (2010). Molecular Aspects Tepkasikul, P., Panya, A. and Valyasevi, R. (2011). Biogenic of Fragrance and Aroma in Rice. Advances in Botanical amine formation in Nham, a Thai fermented sausage, and Research, 56(c), 50-73. the reduction by commercial starter culture, Lactobacillus 207. Vanichtanankul, J., Taweechai, S., Yuvaniyama, J., plantarum BCC 9546. Food Chemistry, 129(13), 846-853. Vilaivan, T., Chitnumsub, P., Kamchonwongpaisan, S. 198. Tran, T., Lee, B.H., Yang, H.S., Chotineeranat, S., Sriroth, K. and Yuthavong, Y. (2011). Trypanosomal Dihydrofolate and Kim, H.J. (2011). Use of starch granules melting to con- Reductase Reveals Natural Antifolate Resistance. ACS trol the properties of bio-flour filled polypropylene and Chemical Biology, 6(9), 905-911. poly(butylene succinate) composites: Physico-chemical properties. Starch-Stärke, doi:10.1002/star.201100025.
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208. Wangchuk, P., Keller, P.A., Pyne, S.G., Taweechotipatr, M., 214. Wongkaew, N., Guajardo, C., Rijiravanich, P., Tonsomboon, A., Rattanajak, R. and Kamchonwongpaisan, Somasundrum, M. and Surareungchai, W. (2011). Use S. (2011). Evaluation of an ethnopharmacologically se- of 3-D plots to avoid mutual interference in bianalyte lected Bhutanese medicinal plants for their major classes ASV determinations: Application to cadmium and lead of Phytochemicals and biological activities. Journal of detection. Talanta, 85(2), 1225-1228. Ethnopharmacology, 137(1), 730-742. 215. Woramongkolchai, N., Supungul, P. and Tassanakajon, 209. Wanitchang, A., Jengarn, J. and Jongkaewwattana, A. A. (2011). The possible role of penaeidin5 from the (2011). The N terminus of PA polymerase of swine-origin black tiger shrimp, Penaeus monodon, in protection influenza virus H1N1 determines its compatibility with against viral infection. Developmental and Comparative PB2 and PB1 subunits through a strain-specific amino Immunology, 35(5), 530-536. acid serine 186. Virus Research, 155(1), 325-333. 216. Yamada, N., Cha-Um, S., Kageyama, H., Promden, W., 210. Wanitchang, A., Patarasirin, P., Jengarn, J. and Tanaka, Y., Kirdmanee, C. and Takabe, T. (2011). Isolation Jongkaewwattana, A. (2011). Atypical characteristics and characterization of proline/betaine transporter gene of nucleoprotein of pandemic influenza virus H1N1 from oil palm. Tree Physiology, 31(4), 462-468. and their roles in reassortment restriction. Archives of 217. Yamada, Y., Yukphan, P., Vu, H.T.L., Muramatsu, Y., Virology, 156, 1031-1040. Ochaikul, D. and Nakagawa, Y. (2011). Subdivision of the 211. Watthanasurorot, A., Jiravanichpaisal, P., Liu, H., Söderhäll, genus Gluconacetobacter Yamada, Hoshino and Ishikawa I. and Söderhäll, K. (2011). Bacteria-Induced Dscam 1998: the proposal of Komagatabacter gen. nov., for Isoforms of the Crustacean, Pacifastacus leniusculus. strains accommodated to the Gluconacetobacter xylinus PLoS Pathogens, 7(6), e1002062. group in the α-Proteobacteria. Annals of Microbiology, 212. Watthanasurorot, A., Söderhäll, K., Jiravanichpaisal, P. doi:10.1007/s13213-011-0288-4. and Söderhäll, I. (2011). An ancient cytokine mediates 218. Yukphan, P., Malimas, T., Muramatsu, Y., Potacharoen, circadian regulation of invertebrate hematopoiesis. W., Tanasupawat, S., Nakagawa, Y., Tanticharoen, M. Cellular and Molecular Life Sciences, 68(2), 315-323. and Yamada, Y. (2011). Neokomagataea gen. nov., with 213. Whankaew, S., Poopear,S., Kanjanawattanawong, Descriptions of Neokomagataea thailandica sp. nov. S.,Tangphatsornruang, S., Boonseng, O., Lightfoot, and Neokomagataea tanensis sp. nov., Osmotolerant D.A. and Triwitayakorn, K. (2011). A genome scan for Acetic Acid Bacteria of the α-Proteobacteria. Bioscience, quantitative trait loci affecting cyanogenic potential of Biotechnology, and Biochemistry, 75(3), 419-426. cassava root in an outbred population. BMC Genomics, 13, 266.
Annual Report 2011 90 List of Patents and Petty Patents
Title Filing Date Ref. No. Country
Patent
Transgenic plants with reduced expression of AMADH2 and elevated levels 6 December 2010 US 12/961398 USA of 2-acetyl-1-pyrroline
Process of preparing polylactic acid-starch composite nanofiber by 9 December 2010 1101000490 Thailand electrospinning
Primers covering infectious hypodermal and hematopoietic necrosis virus 3 February 2011 1101000154 Thailand (IHHNV) genome
The folAthyA knock-out (KO) cells for screening dihydrofolate reductase- 9 February 2011 1101000176 Thailand thymidylate synthase (PfDHFR-TS) inhibitors
Transgenic Plasmodium models for Plasmodium vivax screening 9 February 2011 1101000177 Thailand
Transgenic Plasmodium models for Plasmodium falciparum screening 9 February 2011 1101000178 Thailand
Turbidimeter for detection of viral infection and its method 4 March 2011 1101000330 Thailand
Anti-folate antimalarials with dual-binding modes and their preparation. 10 March 2011 1101000353 Thailand
Formulation of a bioremediating agent for oil degradation 29 April 2011 1101000600 Thailand
One-step bovine embryo sex determination method using multiplex-LAMP 29 April 2011 1101000605 Thailand technique
Cancer-cell inhibiting compound, Xyrrolin; 4-hexyl-2-(hydroxymethyl)-2,5- 2 June 2011 1101000765 Thailand dihydro-1H-pyrroline-3ol
Method to test the mutation of HIV integrase gene 30 June 2011 1101000994 Thailand
Dried phytase encapsulated from a multiple emulsion technique for 14 July 2011 1101001153 Thailand feed industry
Antimicrobial bioplastic film for food packaging and its preparation 21 July 2011 1101001215 Thailand
Biocatalyst for biodiesel production and its application 11 August 2011 1101001495 Thailand
Preparation method and applications of antibody labeled with an enzyme 18 August 2011 1101001600 Thailand encapsulation
Biodegradable thermoplastic starch from starch modified with enzyme from 15 September 2011 1101002053 Thailand Aspergillus niger BCC 7178 and its preparation
Pulp bleaching process without the need of pH adjustment using alkaline- 15 September 2011 1101002054 Thailand tolerant xylanase obtained from metagenomic library of termite gut
Antibody array in a multiple well plate format for multiplex detection of 22 September 2011 1101002173 Thailand bateria and virus in melon seeds
Process to remove organic matters in high-salinity wastewater from sugar- 30 September 2011 1101002384 Thailand mill industry using halophilic microorganisms
Biocontrol agent containing Beauveria bassiana BCC 2660 and Vip3Ag2 30 September 2011 1101002385 Thailand protein for beet armyworm (Spodoptera exigua)
Biocontrol agent containing Beauveria bassiana BCC 2660 and Vip3Aa35 30 September 2011 1101002386 Thailand protein for beet armyworm (Spodoptera exigua)
Petty Patent
Method to induce spore germination in wild mushroom 3 February 2011 1103000114 Thailand
Culture medium to enhance endospore production (formulation 1) 9 June 2011 1103000566 Thailand
National Center for Genetic Engineering and Biotechnology (BIOTEC) 91
Title Filing Date Ref. No. Country
Culture medium to enhance endospore production (formulation 2) 9 June 2011 1103000567 Thailand
Culture medium to enhance endospore production (formulation 3) 9 June 2011 1103000568 Thailand
Culture medium to enhance endospore production (formulation 4) 9 June 2011 1103000569 Thailand
Culture medium to enhance endospore production (formulation 5) 9 June 2011 1103000570 Thailand
Detection of white spot syndrome virus in shrimp using LAMP technique 17 June 2011 1103000606 Thailand and colorimetry
Dried cellulase and xylanase encapsulated from a solution technique for 14 July 2011 1103000688 Thailand feed industry
Dried phytase encapsulated from a solution technique for feed 14 July 2011 1103000689 Thailand industry
Pretreatment process of cassava materials for thermoplastic starch production 22 July 2011 1103000710 Thailand
Enzymatic process for pulp grinding in the paper-making process 28 July 2011 1103000736 Thailand
Colorimetric detection method of methylparathion based on the interaction 4 August 2011 1103000782 Thailand of gold nanoparticle and methylparathion hydrolase
Method for Listeria monocytogenes detection based on antibody array using 18 August 2011 1103000847 Thailand liposome encapsulation conjugated with antibodies and enzyme for naked eye signal detection
Specific primers for shrimp parasite, Enterocytozoon hepatopenaei, and its 9 September 2011 1103000952 Thailand application List of Granted Patents / Petty Patents
Title Granting Date (Petty) Patent No. Country
Patent
BADH2 nucleic acids associated with grain aroma 7 December 2010 US 7847083 USA
Continuous microalgal culture system 3 February 2011 TH 29560 Thailand
Antituberculosis compounds, Hirsutellones A, B, and C 3 February 2011 TH 29561 Thailand
Nucleic acids that enhance the synthesis of 2-acetyl-1-pyrroline in plants 22 July 2011 JP 4786353 Japan and fungi
Petty Patent
Continuous biogas production unit 3 December 2010 TH 5814 Thailand
Cryopreservation of spermatophore of Penaeus monodon* 17 February 2011 TH 5986 Thailand
Sucrose measuring device based on pulse amperometry for sugar mill 1 April 2011 TH 6127 Thailand factory
Sandworm (polychaetes) harvesting device 30 June 2011 TH 6333 Thailand
Seed coating substance* 14 July 2011 TH 6373 Thailand
* Inventions made by researchers from universities with BIOTEC’s research grant.
Annual Report 2011 92
Honors and Awards
Assoc. Prof. Dr. Apichart Vanavichit Dr. Saengchan Senapin Rice Gene Discovery Unit Center of Excellence for Molecular Biology and Genomics of Outstanding Technologist Award 2010 for the work Shrimp on “Enhancing the aroma (2-accetyl-pyrroline) in Thai L’OREAL Thailand-UNESCO for Women in Science rice and sticky rice”, awarded by The Foundation for 2011 Fellowship (Life Science) for her research on the Promotion of Science and Technology under the the “Study of shrimp molecular biology using yeast Patronage of His Majesty the King two-hybrid technique”, awarded by L’OREAL Thailand and the Thai National Commission for UNESCO Prof. Dr. Anchalee Tassankajon Center of Excellence for Molecular Biology and Genomics of Dr. Supawadee Ingsriswang, Mr. SunaiYokwai and Dr. Shrimp Duangdao Wichadakul NRCT Outstanding Researcher 2010 (Agriculture and Bioresources Technology Unit Biology), awarded by the National Research Council Paper titled “LinkinPath: From Sequence to of Thailand Interconnected Pathway” was highlighted on A-IMBN Research, selected by the Asia-Pacific Dr. Siwat Sangsritavong International Molecular Biology Network (A-IMBN) Agricultural Biotechnology Research Unit Award from H.R.H. Princess Maha Chakri Sirindhorn Dr. Suthasinee Somyong and researchers from Cornell for his dedicated efforts to improve dairy farm and University industry, presented at the 2011 National Dairy Fair Genome Institute and Cornell University Paper titled “Comparative genetic analysis of Dr. Taridaporn Buajarern a wheat seed dormancy QTL with rice and Bioresources Technology Unit Brachypodium identifies candidate genes for ABA Wiley-CST Outstanding Publication Award 2010 for perception and calcium signaling” was highlighted the work on “Isoriotins E and F, spirocyclic and bicyclic on A-IMBN Research, selected by the Asia-Pacific hemiacetals from the entomopathogenic fungus International Molecular Biology Network (A-IMBN) Isaria tenuipes BCC 12625”, awarded by Wiley and the Chemical Society of Thailand Dr. Anan Jongkaewwattana, Mr. Asawin Wanitchang and Mr. Juggragarn Jengarn Dr. Piti Amparyup Agricultural Biotechnology Research Unit Center of Excellence for Molecular Biology and Genomics of Paper titled “The N terminus of PA polymerase of Shrimp swine-origin influenza virus H1N1 determines its Young Biochemistry and Molecular Biology Award compatibility with PB2 and PB1 subunits through a 2011, awarded by the Science Society of Thailand strain-specific amino acid serine 186” was highlighted under the Patronage of His Majesty the King on A-IMBN Research, selected by the Asia-Pacific International Molecular Biology Network (A-IMBN) Dr. Jariya Sakayaroj, et al. Bioresources Technology Unit Ms. Thidathip Wongsurawat, Dr. Rungnapa Leelatanawit, Young Scientist Award 2011 for the research project Ms. Natechanok Thamniemdee, Ms. Umaporn Uawi- titled “Butt rot disease in Thailand’s mangroves", setwathana, Dr. Nitsara Karoonuthaisiri, Prof. Piamsak awarded by the UNESCO Man and Biosphere (MAB) Menasveta and Dr. Sirawut Klinbunga Programme Agricultural Biotechnology Research Unit, Center of Excellence for Marine Biotechnology and Chulalongkorn University Dr. Darin Kongkasuriyachai Paper titled “Identification of testis-relevant Medical Molecular Biology Research Unit genes using in silico analysis from testis ESTs Career Development Fellowships (CDFs) in research and cDNA microarray in the black tiger shrimp areas related to infectious diseases of poverty, awarded (Penaeus monodon)” was highlighted on by The UNICEF/UNDP/World Bank/WHO Special A-IMBN Research, selected by the Asia-Pacific Programme for Research and Training in Tropical International Molecular Biology Network (A-IMBN) Diseases (TDR).
Dr. Nitsara Karoonuthaisiri Agricultural Biotechnology Research Unit Marie Curie International Incoming Fellowship (IIF) for her research on “Pathfinder-Rapid and Reliable Detection of Foodborne Pathogens by Employing Multiplexing Biosensor Technology”, awarded by the European Union Framework Program 7
National Center for Genetic Engineering and Biotechnology (BIOTEC) 93
Mr. Pattanop Kanokratana, Ms. Tanaporn Uen- Mr. Yutthana Kingcha gwetwanit, Mr. Ukrit Rattanachomsri, Ms. Benjarat Food Biotechnology Research Unit Bunterngsook Dr. Thidarat Nimchua Dr. Sithichoke Outstanding Oral Presentation Award for the work Tangphatsornruang, Dr. Vethachai Plengvidhya, titled “Whole cell detection of pediocin PA-1/AcH Dr. Verawat Champreda and Dr. Lily Eurwilaichitr producing lactic acid bacteria by MALDI-TOF/TOF mass Bioresources Technology Unit, Genome Institute and Food spectrometry to be used as starter culture in Nham, a Biotechnology Research Unit traditional fermented pork sausage”, presented at JSPS Paper titled “Insights into the Phylogeny and Asian CORE Program (2009-2013), Hanoi Meeting on Metabolic Potential of a Primary Tropical “Next-Generation Bioproduction Platform Leveraging Peat Swamp Forest Microbial Community by Subtropical Microbial Bioresources” , 14-15 October Metagenomic Analysis” was highlighted on 2010, Hanoi, Vietnam A-IMBN Research, selected by the Asia-Pacific International Molecular Biology Network (A-IMBN) Mr. Yutthana Kingcha Food Biotechnology Research Unit Sombat Rukpratanporn and researchers from the Depart- Outstanding Oral Presentation Award for the work ment of Biology, Faculty of Science, Srinakharinwirot titled “Screening of Potential Probiotic Strain of University Lactobacillus plantarum for Use as Nham Starter Center of Excellence for Marine Biotechnology and Culture”, presented at The 22nd Annual Meeting of Srinakharinwirot University the Thai Society for Biotechnology and International Paper titled “The development of loop-mediated Conference on Biotechnology for Healthy Living, 20- isothermal amplification combined with lateral flow 22 October 2010, Trang, Thailand dipstick for detection of Vibrio parahaemolyticus” was highlighted on A-IMBN Research, selected by the Asia- Ms. Aritsara Jaruwat Pacific International Molecular Biology Network (A-IMBN) Medical Molecular Biology Research Unit Poster Presentation Award (first runner-up) for the work Ms. Jarunee Vanichtanankul, Ms. Supannee Taweechai, Dr. titled “Schiff’s base formation of Lys-237 and PLP reveals Penchit Chitnumsub, Dr. Sumalee Kamchonwongpaisan, in crystal structure of Serine hydroxymethyltransferase Prof. Dr. Yongyuth Yuthavong, Dr. Jirundon Yuvaniyama from Plasmodium falciparum”, presented at The 6th and Dr. Tirayut Vilaivan International Symposium of the Protein Society of Medical Molecular Biology Research Unit, Mahidol University Thailand, 30 August - 2 September 2011, Bangkok, and Chulalongkorn University Thailand Paper titled “Trypanosomal dihydrofolate reductase reveals natural antifolate resistance” was selected and Ms. Chayaphat Wongsombat evaluated by Vivian Cody, a Member of the Faculty Medical Molecular Biology Research Unit of 1000 (F1000), which places the work in the F1000 Poster Presentation Award (second runner-up) library of the top 2% of published articles in biology for the work titled “Identifying mRNA targets of P. and medicine. falciparum RNA binding proteins”, presented at The 6th International Symposium of the Protein Society Ms. Benchaporn Buaban, Dr. Hiroyuki Inoue, Dr. Shinichi of Thailand, 30 August - 2 September 2011, Bangkok, Yano, Dr. Sutipa Tanapongpipat, Dr. Vasimon Ruanglek, Thailand Dr. Verawat Champreda, Dr. Rath Pichyangkura, Dr. Sirirat Rengpipat and Dr. Lily Eurwilaichitr Ms. Nattida Suwanakitti Bioresources Technology Unit, National Institute of Advanced Medical Molecular Biology Research Unit Industrial Science and Technology (AIST) and Chulalongkorn Poster Presentation Award (consolation prize) for University the work titled “A novel bacterial surrogate system 2011 Excellent Paper Award for the paper titled for antifolate screening against malaria and other “Bioethanol production from ball milled bagasse parasitic diseases”, presented at The 6th International using an on-site produced fungal enzyme cocktail Symposium of the Protein Society of Thailand, 30 and xylose-fermenting Pichia stipitis”, awarded by the August - 2 September 2011, Bangkok, Thailand Society for Biotechnology, Japan
Annual Report 2011 94
Ms. Charuwan Chuaseeharonnachai, Mr. Veera Sri-Indra- Dr. Piti Amparyup sutdhi, Ms. Kaoru Yamaguchi Dr. Akira Nakagiri, Dr. Izumi Center of Excellence for Molecular Biology and Genomics of Okane and Mr. Nattawut Boonyuen Shrimp Bioresources Technology Unit and National Institute of Dissertation Award 2010 (Agriculture and Biology) Technology and Evaluation (NITE) for “Development of genetic markers for increasing Oral Presentation Award for the work titled “Diversity production efficiency of the tropical abalone Haliotis and systematics of aero-aquatic fungi in two major asinina in Thailand”, awarded by National Research tropical rainforests, Thailand”, presented at The 1st Council of Thailand Conference on Taxonomy and Systematics in Thailand, 2-4 May 2011, Phitsanulok, Thailand Dr. Honglada Thoetkiattikul Bioresources Technology Unit Mr. Chanwit Suriyachadkun Dissertation Award 2010 (Agriculture and Biology) for Bioresources Technology Unit “Expression and functional studies of the Microplitis Oral Presentation Award for the work titled “Diversity demolitor Bracovirus IkBLIKEGene family)” awarded by of actinomycetes in the family Streptosporangiaceae National Research Council of Thailand in Thailand”, presented at The 1st Conference on Taxonomy and Systematics in Thailand, 2-4 May 2011, Dr. Srung Smanmoo Phitsanulok, Thailand Bioresources Technology Unit First runner-up for the work on “The synthesis of highly Mr. Artit Khonsanit, Ms. Kanoksri Tasanathai, Ms. Kanoksri sensitive fluorescent and colorimetric chemosensor Tasanathai, Ms. Donnaya Thanakitpipattana, Ms. Wasana for detection of Hg(II) ion in aqueous environment Noisripoom and Dr. Janet J. Luangsa-Ard and living cells”, at the Environmental Research Bioresources Technology Unit Competition: Sustainable Development organized by Poster Presentation Award for the work titled the Department of Environmental Quality Promotion “Diversity of Cordyceps nipponica affecting different hosts in Thailand”, presented at The 1st Dr. Verawat Champreda, Mr. Ukrit Rattanachomsri and Conference on Taxonomy and Systematics in students from Thammasart University International MBA Thailand, 2-4 May 2011, Phitsanulok, Thailand Program Bioresources Technology Unit Ms Wipaporn Ngaemthao, Mr. Chanwit Suriyachadkun, Ms. First Prize Winner for the work on “ENZstrip: test Suwanee Chunhametha and Dr. Vichien Kitpreechavanich strips containing in-gel crosslink substrate for Bioresources Technology Unit and Kasetsart University detectingenzyme activity for industrial and academic Poster Presentation Award for the work titled research use”, presented at The NSTDA Idea to Product “Sphearisporangium krabiensis sp. nov., Competition: Fast Track to Commercialization 2010, isolated from Thailand”, presented at The 1st 4-5 November 2010 Pathum Thani, Thailand Conference on Taxonomy and Systematics in Thailand, 2-4 May 2011, Phitsanulok, Thailand Dr. Mongkon Audtho and students from Thammasart University International MBA Program Mr. Winai Chaipitakchonlatarn, Ms. Mika Miyashita, Dr. Bioresources Technology Unit Pattaraporn Rattanawaree, Mr. Taweesak Malimas, Mr. Second runner-up and Rocket Pitch awards for Somphop Boonpayung, Ms. Yuki Muramatsu, Dr. Yas- the work on “The Production and Utilization of uyoshi Nakagawa, Dr. Ken-ichiro Suzuki, Dr. Somboon Vegetative Insecticidal Protein (VIP) for Controlling Tanasupawat, Ms. Wanchern Potacharoen Insect Pest”, presented at The NSTDA Idea to Product Bioresources Technology Unit, Chulalongkorn University and Competition: Fast Track to Commercialization National Institute of Technology and Evaluation (NITE) 2010, 4-5 November 2010 Pathum Thani, Thailand Poster Presentation Award for the work titled “Taxonomic and ecological study of lactic acid Dr. Thidarat Nimchua bacteria from fermented foods in Central and Bioresources Technology Unit South-Northern part of Thailand”, presented at Best Presentation Award for the work on “ENZbleach: The 1st Conference on Taxonomy and Systematics An alkaline-tolerant enzyme for pulp bleaching in Thailand, 2-4 May 2011, Phitsanulok, Thailand process”, presented at NSTDA Investors’ Day, 22 September 2011 in Pathum Thani, Thailand Ms. Donnaya Thanakitpipattana, Ms. Kanoksri Tasanathai, Mr. Rungpet Ridkaew and Dr. Janet J. Luangsa-Ard Bioresources Technology Unit Poster Presentation Award for the work titled “A new species of Ophiocordyceps halabalaenses in Hala-Bala Wildlife Sanctuary, Southern Thailand”, presented at The 1st Conference on Taxonomy and Systematics in Thailand, 2-4 May 2011, Phitsanulok, Thailand
National Center for Genetic Engineering and Biotechnology (BIOTEC) 95 Executives and Management Team
Executive Board
Advisors Prof. Sanit Aksornkoae President, Thailand Environment Institute (TEI) Prof. Naksitte Coovattanachai Director of the Royal Golden Jubilee PhD Program, Thailand Research Fund (TRF) Dr. Sermpol Ratasuk Expert, TEAM Consulting Engineering and Management Co., Ltd. (TEAM) Dr. Sakarindr Bhumiratana President, King Mongkut’s University of Technology Thonburi
Chairman Prof. Sujin Jinahyon President, Naresuan University
Vice Chairman Dr. Thaweesak Koanantakool President, National Science and Technology Development Agency (NSTDA)
Members Prof. Amaret Bhumiratana Faculty of Science, Mahidol University Prof. Narongsak Chaiyabutr Faculty of Veterinary Science, Chulalongkorn University Mr. Somchai Charnnarongkul Director General, Department of Agriculture Dr. Vichai Chokevivat Director, Institute for Development of Human Research Protection Dr. Somsak Chunharas Secretary General, National Health Foundation Mr. Pornsil Patchrintanakul Vice President, Charoen Pokphand Group Deputy Secretary General, Thai Chamber of Commerce Mrs. Vipajaree Putthamilinprateep Bureau of the Budget Prof. Sawasd Tantaratana Director, Thailand Research Fund (TRF) Prof. Peerasak Srinives Faculty of Agriculture, Kasetsart University Prof. Prapon Wilairat Faculty of Science, Mahidol University Dr. Kanyawim Kirtikara Executive Director, BIOTEC Ms. Dussadee Siamhan Deputy Executive Director, BIOTEC
Annual Report 2011 96
International Advisory Board
Chairman Prof. Ken-ichi Arai Professor Emeritus, The University of Tokyo, JAPAN
Members Dr. Jill Conley Director, International and Precollege Science Education Programs Howard Hughes Medical Institute (HHMI), USA Prof. Douglas Hilton Director, Walter and Eliza Hall Institute, AUSTRALIA Dr. Ming-Chu Hsu Chairman & CEO, TaiGen Biotechnology Co., Ltd., TAIWAN Dr. Martin Keller Associate Laboratory Director of Biological and Environmental Sciences, Oak Ridge National Laboratory (ORNL), USA Prof. Gerald T. Keusch Professor of International Health and of Medicine, Boston University School of Public Health, USA Prof. Lene Lange Vice Dean for Faculty of Engineering, Science and Medicine, Aalborg University, DENMARK Prof. Sangkot Marzuki Director, Eijkman Institute for Molecular Biology, INDONESIA Dr. Jean-Marcel Ribaut Director, Generation Challenge Program (GCP)
Management Team
Dr. Kanyawim Kirtikara Dr. Lily Eurwilaichitr Executive Director Director, Bioresources Technology Unit
Ms. Dussadee Siamhan Dr. Somvong Tragoonrung Deputy Executive Director Director, Genome Institute
Dr. Suvit Tia Dr. Sumalee Kamchonwongpaisan Deputy Executive Director Director, Medical Molecular Biology Research Unit
Ms. Kruawan Potisombat Dr. Sirawut Klinbunga Assistant Executive Director Director, Agricultural Biotechnology Research Unit
Dr. Wonnop Visessanguan Director, Food Biotechnology Research Unit
National Center for Genetic Engineering and Biotechnology (BIOTEC)