BIOTECHNOLOGY IN NEPAL

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BIOTECHNOLOGY IN NEPAL

• A SYNOPSIS
• Professor Dr. Sarbajna Man Tuladhar
• Research Centre for Applied Science and
  Technology (RECAST)
• Tribhuvan University, Kirtipur, Kathmandu, Nepal.
• E-mail phoenix_at_wlink.com.np turecast_at_mail.com.np

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BIOTECHNOLOGY IN NEPAL A SYNOPSIS

• Country paper presented at Expert Group Meeting
  on Biotechnology Information Network for Asia
  (BINASIA) 24-25 January 2006, Bangkok, Thailand
  organized by UN APCTT, KRIBB in cooperation with
  BIOTEC, Thailand.

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1. INTRODUCTION

• Biotechnology has been around us for centuries.
  In industrialized countries, modern biotechnology
  has become well advanced.
• In the last decade of the 20th century, Nepal
  witnessed several biotechnology-derived products
  in the market
• Human health care products
• Seeds and plants of high yielding variety of
  agricultural and horticultural crops
• Food products and drugs
• Improved livestock
• Animal health care products
• THEY ENTERED INTO THE COUNTRY FROM THE
  NEIGHBOURING AND THE DEVELOPED COUNTRIES.

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• MODERN BIOTECHNOLOGY HAS ENTERED INTO NEPAL LIKE
  BANDWAGON AND EVERYBODY IS JUMPING ONTO IT.
• IN FACT BIOTECHNOLGY PROGRAMMES ARE RUNNING
  WITHOUT ANY POLICIES AND PLANS.
• MODERN BIOTECHNOLOGY IN NEPAL IS STILL IN A STATE
  OF INFANCY.

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2. THE CONCERNED MINISTRIES

• Ministry of Environment, Science and Technology
• Ministry of Forest and Soil Conservation
• Ministry of Agriculture and Co-operative
• Ministry of Health
• Ministry of Education and Sports
• Ministry of Industry, Commerce and Supply

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3. PRESENT STATUS OF DIFFERENT AREAS OF
BIOTECHNOLOGY
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3.1 PLANT TISSUE CULTURE

• MAJOR BIOTECHNOLOGY ACTIVITY OF PUBLIC AS WELL AS
  PRIVATE SECTORS.
• PUBLIC LABORAORIES DEVELOPED PROTOCOLS FOR IN
  VITRO PROPAGATION OF DIFFERENT PLANT SPECIES.
• PRIVATE LABORATORIES PRODUCED ELITE PLANTS
  COMMERCIALLY USING PROTOCOLS DEVELOPED BY PUBLIC
  LABORATORIES.

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3.1.1 PUBLIC SECTOR INITIATIONS

• PROMINENT LABORATORIES
• DEPARTMENT OF PLANT RESOURCES (DPR)
• First tissue culture laboratory established at
  Godawari to initiate and promote plant tissue
  culture in 1976.
• Developed protocols for tissue culture
  propagation of more than 100 plant species
  including orchids, agricultural and horticultural
  trees, ornamental plants, and bamboo.

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• Most important achievement is the development of
  sand rooting technique the tissue-cultured
  micro shoots can be rooted in the non-sterile
  sand.
• Sand rooting technique is easy, cheap, less time
  consuming, economic, conveniently done in
  nurseries, can easily be established for field
  plantation.
• Provides tissue culture training for
  entrepreneurs. Consequently four private plant
  tissue culture factories have been established.

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• 2. NEPAL AGRICULTURE RESEARCH COUNCIL (NARC)
• Potato Research Program (PRP) with the assistance
  of Swiss Development Corporation has been
  producing virus-free pre-basic seeds of potato at
  its tissue culture laboratories and green house
  facilities at Khumaltar.
• DIVISION OF AGRICULTURE BOTANY of NARC has
  initiated some works on
• anther culture of rice and wheat,
• germ plasm conservation and diagnostic
  facilities
• using Polymerase Chain Reaction (PCR)
  technology.
• Established tissue culture laboratories at
  regional agricultural research stations at
  Dhankuta (Eastern development region) and Lumle
  (Mid-western development region) to promote
  micropropagaton.

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• 3. HORTICULTURE DEVELOPMENT PROJECT/ DEPARTMENT
  OF AGRICULTURE
• Tissue culture propagation of apple rootstocks
• Micro grafting of sweet orange (JUNAR) for the
  production of virus-free and greening free
  plants.
• The facilities remain unused at present

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• 4. INSTITUTE OF AGRICULTURE AND ANIMAL SCIENCES
  (IAAS) / T. U.
• Established tissue culture facilities with the
  grant assistance from Rockefeller Foundation.
• Assisting thesis works for M. Sc. Degree.

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3.1.2 UNIVERSITY SECTOR ACTIVITIES

• CENTRAL DEPARTMENT OF BOTANY/ TRIBHUVAN
  UNIVERSITY
• Tissue culture propagation of different medicinal
  plants.
• Anther culture.
• Rhizobium inoculation in leguminous plants

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3.1.3. PRIVATE SECTOR INITIATIONS

• BOTANICAL ENTERPRISES PVT. LTD.
• In vitro propagation of many species of orchid,
  potato, fodder, Chrysanthemum, Gerbera, African
  violet, Lily etc.
• Export directly to The Netherlands.
• NEPAL BIOTECH NURSERY
• Produce banana, orchid, and ornamental plants by
  tissue culture and non-sterile sand rooting
  technique.

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• 3. RESEARCH LABORATORY FOR AGRICULTURE
  BIOTECHNOLOGY AND BIOCHEMISTRY (RALBB)
• Tissue culture propagation for pine, Artocarpus,
  Brassica.
• Anther culture of cold tolerant rice
• Modest facilities for DNA work by PCR technology
  and enzyme analysis.
• Facilities used for research and teaching.

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• 4. MICROPLANTS NEPAL PVT. LTD.
• Commercial propagation of banana, orchid, lily,
  mums etc.
• 5. GREEN RESEARCH AND TECHNOLOGY (GREAT)
• Developing virus testing and elimination
  facilities on horticultural crops such as potato,
  citrus, banana, cardamom, strawberry, and some
  ornamental plants using tissue culture
  techniques.
• Has modest screen-house facility for indexing
  against citrus greening disease.

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• Cleaned two potato cultivars of Bangladesh namely
  Petronese and Multa from potato viruses such as
  PVX and PVY.
• Cleaned three leading cultivars of large cardamom
  (Amomum subutalum) namely Ramshahi, Golshahi, and
  Dambarshahi from Chhirke and Furke viruses.
• Virus-free plants derived from meristem culture
  are available for further multiplication.

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• HIMALAYAN BOTANICAL RESEARCH CENTRE (HIMBORCE)
• Established infrastructure for micropropagation
  of medicinal and ornamental plants indigenous to
  Nepal.
• Embarking upon micropropagation of Gladiolus.

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3.2 BIOFERTILIZERS

• ROYAL NEPAL ACADEMY OF SCIENCE AND TECHNOLOGY
  (RONAST)
• Developed Rhizobium inoculant technology for
  pulses (lentils).
• Developed microbial inoculant technology for
  composting.
• Use of ecto and endomycorrhiza in conservation of
  forest soils.
• Quality production of organic fertilizers.

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• 2. DIVISION OF SOIL SCIENCE / NARC
• Production and distribution of legume inoculant
  packets for more than a decade.
• Distributed 2627 Rhizobium inoculant packets to
  the farmers for important pulse and pasture
  crops.
• 3. DIVISION OF PLANT PATHOLOGY / NARC
• Engaged in spawn production technology for
  mushroom culture using two genera namely Agaricus
  and Pleurotus.

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• RESEARCH CENTRE FOR APPLIED SCIENCE AND
  TECHNOLOGY (RECAST) / T. U.
• Improvement of traditional composting technology.
• An integrated approach for the utilization of
  municipality solid wastes.

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• 5. CENTRAL DEPARTMENT OF
• MICROBIOLOGY / T. U
• AND
• 6. CENTRAL DEPARTMENT OF CHEMISTRY / T. U.
• Impact of industrial waste on land and water
  pollution.

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3.3 BIOPESTICIDES

• ROYAL NEPAL ACADEMY OF SCIENCE AND TECHNOLOGY
  (RONAST)
• Several isolations of Bacillus thuringiensis (Bt)
  from Nepalese soils for biological control of
  Lepidoptera caterpillar in crucifer vegetables.
• Field trial of indigenous pesticide plants like
  Eupatorium adenophorum, Lantana camara,
  Azadirachta indica etc. in collaboration with
  Department of Agriculture (Ministry of
  Agriculture).

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• 2. DIVISION OF ENTOMOLOGY / NARC
• Reported efficacy of Acorus calamus (Root
  powder), neem oil (Azadiractin, 0.15), and neem
  seed extract (Azadiracta monotetala) against
  storage insect like Sitophilus zeamais.
• 3. RESEARCH CENTRE FOR APPLIED SCIENCE AND
  TECHNOLOGY / T. U.
• Survey of plant pesticides for grain storage.

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3.4 MEDICAL BIOTECHNOLOGY

• THE LEPROSY MISSION, NEPAL
• Determination of cytokines by ELISA for blood
  immune response studies in leprosy patients.
• DNA diagnosis of Mycobacterium leprae by PCR.
• CENTRAL VETERINARY LABORATORY
• Vaccine production against PPR (Pestes des
  Petites Ruminant) using tissue culture.

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• NEPAL AGRICULTURE RESEARCH COUNCIL (NARC)
• Performing embryo transfer and artificial
  insemination in cattle.
• EVEREST BIOTECH PVT. LTD.
• Production of specific antibody required of Human
  Genome Project, England.

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• 5. CENTRAL DEPARTMENT OF CHEMISTRY/
• T.U.
• Antibacterial, antifungal, and cytotoxic
  activities of medicinal plants.
• Study of specific inhibitors for proteolytic
  enzymes.
• Taxol from endophytes of Himalayan conifers.

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• 6. CENTRAL DEPARTMENT OF MICROBIOLOGY / T. U.
• Antimicrobial activities of some plant extracts.
• 7. RESEARCH CENTRE FOR APPLIED SCIENCE AND
  TECHNOLOGY / T. U.
• Bioprospecting of ethno medicinal plants of Nepal
  for the conservation of biological and cultural
  resources.

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3.5 DISEASE DIAGNOSTICS

• ROYAL NEPAL ACADEMY OF SCIENCE AND TECHNOLOGY
  (RONAST)
• Diagnosis of Asian citrus greening diseases or
  Huanglongbing of citrus using PCR based
  technique.
• Diagnosis of Citrus tristeza virus disease using
  Double Antibody Sandwich Enzyme Linked
  Imunosorbent Assay (DAS-ELISA) technique.

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• INSTITUTE OF AGRICULTURE AND ANIMAL SCIENCES
  (IAAS) / T. U.
• Diagnosis of viral and bacterial diseases in rice
  using PCR technology.

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• GREEN RESEARCH AND TECHNOLOGY (GREAT)
• Use of PCR technology for routine diagnosis of
  citrus-greening disease (Huanglongbing).
• Provides training on virus testing using
  DAS-ELISA and biological indicator plants for the
  diagnosis of Huanglongbin on citrus.

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3.6 BIOENERGY AND BIOCONVERSION TECHNOLOGY

• RESEARCH CENTR FOR APPLIED SCIENCE AND TECHNOLOGY
  (RECAST) / T. U.
• Improvement of biogas production by using weeds
  such as Eupatorium adenophorum and water
  hyacinth.
• Biofuel from the Physic nut (Jatropha curcas L)
  for running diesel engine.

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3.7 MOLECULAR CHARACTERIZATION

• NEPAL AGRICULTURE AND RESEARCH COUNCIL (NARC)
• Genetic diversity analysis of Fagopyrum spp.
  (wild sweet and bitter buckwheat) using Randomly
  Amplified Polymorphic DNA (RAPD).
• Genetic diversity analysis of isozymes in
  indigenous crops like rice, finger millet,
  barley, pigeon pea, buckwheat, taro, cucumber,
  sponge gourd, citrus, and Swertia spp.

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• ROYAL NEPAL ACADEMY OF SCIENCE AND TECHNOLOGY
  (RONAST)
• Initiated research work on molecular
  characterization of medicinal plants like Swertia
  chirata, Camellia sinensis etc. and
  microorganisms such as Bacillus thuringiensis.
• RESEARCH CENTRE FOR APPLIED SCIENCE AND
  TECHNOLOGY (RECAST)
• Selection of appropriate genotypes of Mulberry
  and exploitation of genetic yield potential of
  common buckwheat ecotypes.

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3.8 FERMENTATION

• MAJOR TRADITIONAL BIOTECHNOLOGY AND NATIONAL
  HERITAGE OF NEPAL.
• SIGNIFICANT ANCIENT BIOTECHNOLOGIES STILL
  SURVIVING IN NEPAL ARE CONFINED TO
• COTTAGE INDUSTRY SCALE
• OR
• INDIVIDUAL FAMILY CONSUMPTION ARE
• Fermentation and distillation of characteristic
  alcoholic drinks from millet (KODO), rice, brown
  sugar (SAKHAR) by using indigenous yeast.
• The traditional process of making yeast is still
  a closely guarded secret.

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• Fermentation of leafy vegetables into GUNDRUK,
  the favourite food specialties of rural Nepalese.
• GUNDRUK from the leaves of mustard, radish or
  cauliflower.
• SINKI from radishes cut into pieces.
• SINAMANI from the whole radishes without their
  leaves cut out.
• Browning of rice into HAKUWA or HAKUJAKI
  (literally meaning black rice) by indigenous
  fermentation.

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• Production of jute (NALU) from the bark of the
  hemp through fermentation process.
• Production of yogurt (DAHI) and hard cheese
  (CHHURPI).
• Dungs used as fertilizers and dung cakes used as
  fuel for cooking.
• Interesting to note that traditional
  biotechnology is being conserved predominantly by
  womenfolk.

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3.9 PRACTICAL APPLICATION OF BIOTECHNOLOGY
PRODUCTS/ PROCESSES

• 1. COMMERCIAL / SEMI-COMMERCIAL
• Tissue culture raised plantlets of sisso tree,
  orchids, medicinal plants, ornamental plants, and
  virus free banana and citrus.
• COTTAGE INDUSTRY LEVEL
• Characteristic alcohol from millet (KODO), rice,
  and brown sugar (SAKHAR).
• Fermented vegetables GUNDRUK, SINKI, SINAMANI.
• Yogurt (DAHI) and Hard Cheese (CHHURPI).

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3.10 BIOTECHNOLOGY IN ACADEMIA

• AT UNDERGRADUATE LEVEL
• Biotechnology is taught only as a small part of
  the course unit.
• AT GRADUATE / POST-GRADUATE LEVEL
• Central Department of Botany, T. U. M. Sc.
  Thesis works in tissue culture.
• Central Department of Microbiology, T. U. M.
  Sc. thesis works in agriculture biotechnology,
  medical biotechnology, environmental
  biotechnology, genomics, and proteomics.

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• Institute of Agriculture and Animal Sciences, T.
  U. thesis works in diagnosis of viral and
  bacterial diseases in rice using PCR technology.
• Two year M. Sc. in Biotechnology opening soon in
  Tribhuvan University, the oldest and largest
  university of Nepal.
• Kathmandu University B. Sc. with specialization
  in Biotechnology.
• Purbanchal University B. Sc. with
  specialization in Biochemistry (with more
  emphasis on Biology).
• Pokhara University B. Sc. with specialization
  in Biochemistry (with more emphasis on Chemistry).

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3.11 HUMAN RESOURCES

• Persons specialized in biotechnology are only
  nominal.
• Most biotechnology activities are due to
  personnel specialized in Agriculture and Botany.
• 3.12 NATIONAL ORGANISM BANK
• Established in 1985 at Royal Nepal Academy of
  Science and Technology.
• Objectives Isolation, identification, and
  maintenance of the microorganisms (fungi and
  bacteria).
• Acts as a depository to supply authentic
  microbial cultures for the use in various
  research activities of research organizations,
  universities, and industries. This is an attempt
  to conserve and utilize microbial genetic
  resources of the country.

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3.13 BIOTECHNOLOGY INFORMATION NETWORK

• Biotechnological research findings and activities
  are communicated through occasional papers and
  annual reports published by respective
  organizations or national journals.
• Ministry of Environment, Science and Technology
  plans to develop the information networking in
  Biotechnology.
• A resource centre as the focal point of SAARC
  Technical Committee on Science and Technology
  will be developed in due course of time.

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3.14 REGULATORY PROCEDURES

• Nepal has signed the Cartagena Protocol on
  Biosafety on March 2, 2001.
• Focal point for the Biosafety Protocol Chief of
  the Environment Division in the Ministry of
  Forests and Soil Conservation.
• Ministry of Forests and Soil Conservation has
  enforced BIOSAFETY REGULATIONS 2062 BS throughout
  the Kingdom of Nepal on and from May 25, 2005.

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• OTHER PERTINENT LEGISLATIONS
• - Export Import (Control) Act, 2013 BS.
• - Food Act, 2023 BS.
• - Plant Protection Act, 2029 BS
• - Livestock Feed Products Act, 2033 BS
• - Medicine / Drug Act, 2035 BS
• - Seed Act, 2045 BS
• - Livestock Health and Livestock service act,
• 2055 BS

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3.15 REGULATORY MECHANISM

• IMPLEMENTING AGENCY
• Ministry of Forests and Soil Conservation
• Department of Livestock, Animal Quarantine
  Section and 25 Quarantine check posts / Ministry
  of Agriculture and Cooperatives.
• Plant Quarantine Office, Department of
  Agriculture/ Ministry of Agriculture and
  Cooperatives
• Department of Food Research and Quality Control

• RESPONSIBILITY
• National Focal Point for the Biosafety Protocol.
• For enacting The Livestock Health and Livestock
  Service Act.
• For enacting The Plant Protection Act and The
  Seed Act.
• For checking whether any food products are
  polluted or of quality below the permissible
  standard for human consumption.

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• Department of Drug Administration / Ministry of
  Health
• Department of Industry / Ministry of Industry,
  Commerce and Supply
• Department of custom / Ministry of Finance

• For issuing permission for production, import,
  export, and clinical trial of new medicines.
• For issuing license to establish industry and for
  export and import of goods.
• For issuing of custom clearance of goods only
  after seeking recommendation or permission letter
  from the concerned Departments.

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• For dealing with the issues of the LMOs / GMOs,
  capacity of the above-mentioned ministries should
  be increased.
• CAPACITY BUILDING refers to
• Human resources development
• Infrastructure (laboratory and equipments)
  development
• Development of administrative mechanism
• Networking of concerned ministries
• Formulation of legislation

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3.16 INTERNATIONAL AGREEMENTS

• Nepal has always expressed its solidarity and
  commitment toward several international efforts
  related to the environment conservation.
• Consequently it has become a signatory to a
  number of international legal instruments.

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• LEGAL INSTRUMENT
• Plant Protection Agreement for the South-East
  Asia and Pacific Region.
• Convention on Wetlands of International
  Importance Especially waterfowl Habitat.
• Convention for the Protection of the World
  Cultural and Natural Heritage.
• Convention on International Trade in Endangered
  Species of Wild Fauna and Flora.
• World Conservation Strategy
• World Charter for Nature

• DATE SIGNED
• 27 February 1956
• 2 February 1971
• 23 November 1972
• 3 March 1973
• 1980
• 1982

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• International Tropical Timber Agreement.
• Agreement on Network of Aquaculture Centres in
  Asia and the Pacific.
• Conservation on Biological Diversity.
• The Cartagena Protocol on Biosafety.
• World Trade Organization (WTO) member 147th.

• 18 November 1983
• 8 January 1988
• 22 May 1922
• 2 March 2001
• 23 April 2004

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4. MAJOR CONSTRAINTS

• Acute lack of national programme on
  biotechnology.
• Biotechnology programmes are usually dependent
  on the initiative of a few foreign-trained
  scholars and enthusiast scientists at home.
• National Biotechnology Policy is non-existent as
  yet. Draft of the Policy prepared by Ministry of
  Environment, Science and Technology has been
  years in gestation in the Cabinet.
• Lack of qualified and properly trained research
  and development manpower the limiting factor
  both for private and government sectors.
• The availability and quality of infrastructures
  relevant to biotechnology research is negligible.
• Lack of public sector-private enterprise links,
  university-industry interaction, and
  university-university cooperation.

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5. PROSPECTS

• Despite serious handicaps, the prospects for
  growth of biotechnology in Nepal are numerous.
• Biotechnology potentials worth mentioning are
• AGRICULTURE / FOOD SECTOR
• Plant tissue culture
• Biofertilizer
• Biopesticide
• Animal feeds and supplements from agricultural
  products
• INDUSTRY SECTOR
• Food and industrial enzymes
• Fermentation products
• Bioenergy, biofuel and bioconversion

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• HUMAN HEALTH CARE SECTOR
• Development of rapid diagnostic kits and
  techniques
• Production of culture media
• Production of monoclonal antibodies
• Vaccines against malaria, measles, diarrhoea,
  jaundice, cholera etc.
• Therapeutic agents from local plant resources,
  for example, anticancer / antitumour drugs,
  sweeteners, dyes, flavours, fragrances etc.

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• ANIMAL HEALTH CARE SECTOR
• Veterinary diagnostics and vaccines
• Immunology
• Embryo transfer
• MINING SECTOR
• Mineral leaching is particularly important in the
  context of Nepal having several mineral deposits
  scattered all over the country in relatively
  lower volume of mineral deposits.

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• DNA TECHNOLOGY
• Identification and isolation of genes for use in
  molecular hybridization.
• DNA fingerprinting for forensic, diagnosis of
  genetic disease, paternity analysis, immigration,
  and citizenship record.
• Studies on polymorphism and molecular mapping of
  endangered and commercial crops.
• Development of diagnostic kits.
• Selection in breeding programme.
• AQUACULTURE
• Feeds
• Diagnostics

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• HUMAN RESOURCES DEVELOPMENT SECTOR
• Introduction of biotechnology and bioinformatics
  at the graduate and post graduate levels in
  universities.
• More Ph. D.s at home and abroad.
• Academic collaboration with leading universities
  in the neighbour and abroad.
• Develop programmes to foster biotechnology
  entrepreneurship.

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• ENVIRONMENT AND BIODIVERSITY
• Biological control agents
• Urban waste management
• Industrial waste treatment
• Monitoring tools and biosensors

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6.POSSIBLE ROLE OF BINASIA

• A SERIOUS CONSTRAINT IS THE LACK OF INFORMATION
  ON BIOTECHNOLOGIES
• NO INSTITUTION HAS DEVELOPED CAPABILITY IN
  INFORMATION COLLECTION
• RETRIEVAL DISSEMINATION
• THEREFORE IT RELIES ON INDUSTRIALISED COUNTRIES
  FOR SPECIALIZED INFORMATION

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SOLUTION TO INFORMATION SHORTAGE

• to conduct a thorough literature search on the
  existing information, which should then be stored
  in the institutes having the capability to manage
  information and make it easily accessible to
  biotechnology researchers.
• OR
• to set up a biotechnology information centre and
  establish a computer link with institutions in
  the industrialized countries. However this
  depends on the willingness of the institutions in
  the North to share their biotechnological
  information with those in the South.
• PIVOTAL ROLE OF BINASIA
• MAKE AVAIL OF PUBLICATIONS WITH FULL TEXT

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• T H A N K Y O U