FNCA 2006 WORKSHOP ON BIOFERTILIZER
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Attachment III

IV. Comparison on Effects of Sterilization of Biofertilizer Carriers by Irradiation and Autoclaving


1.Sterilization of carriers is important for the maintenance of biofertilizer quality.

2.There are many methods for sterilization. In this workshop, irradiation and autoclave methods were compared.

3.The results are as in the following:

3-1.Irradiation

  1. Positive effects
    No drastic change in chemical and physical properties, suitable for mass production, less time consuming, less labour, suitable for various forms of carrier materials, complete sterilization at proper doses, simple handling of the carriers and enhanced survival rate for Rhizobium and Azospirillum

    *Supporting tables and figures

    Table 1. Effects of autoclaving and gamma irradiation on the physicochemical properties of soybean cake and perilla oil meal by Byun et al . 1988

    Samples

    Treatments

    Crude
    protein (%)

    TBA
    (mole g-1)

    Color parameters

    Lightness
    (L)

    Redness
    (a)

    Yellowness
    (b)

    △ E

     Soybean cake

    Control

    46.3

    1.48x10 -5

    70.8

    3.8

    18.3

    0.0

    5 kGy

    46.5

    1.81x10 -5

    68.4

    4.1

    18.0

    2.4

    10 kGy

    46.6

    1.88x10 -5

    69.0

    4.4

    17.6

    2.0

    Autoclaving

    46.0

    1.56x10 -5

    62.0

    7.0

    18.2

    9.3

     Perilla oil meal

    Control

    46.7

    1.81x10 -5

    37.2

    7.4

    12.6

    0.0

    5 kGy

    46.6

    1.99x10 -5

    37.0

    6.9

    12.8

    0.6

    10 kGy

    46.8

    2.06x10 -5

    37.0

    7.5

    12.8

    0.2

    Autoclaving

    46.4

    1.84x10 -5

    35.4

    7.3

    12.0

    1.8

    △E means color difference.
    It was detected using the color and color difference meter (Model ND-1001 DP) by the researchers.

    Figure 1. Cost of sterilization per biofertilizer package by Gandanegara , Indonesia

    Autoclave

    Irradiation

    Autoclave

    Irradiation

    Container size:
    3 kg in one time
    Box Size : 30 x 30 x 58 cm
    Package Nos : 900 pcs
    25kg for irradiation box
    16 boxes (400kg) in one time

    Table 2. Survival of Rhizobium vigna in carriers sterilized by irradiation and by autoclave by Le Thi Thanh Thuy et.al , Vietnam

    Storage time

    Rhizobium densities (cfu/g)

    irradiation

    autoclave

    0 h

    1,20 x 10 9 1,32 x 10 9

    2 weeks

    9,20 x 10 9 1,00 x 10 9

    1 month

    7,40 x 10 9 2,54 x 10 9

    2 months

    7,80 x 10 9 1,81 x 10 9

    3 months

    2,00 x 10 9 1,56 x 10 9

    6 months

    3,00 x 10 9 1,33 x 10 8
    Rhizobium strains 132

    Table 3. Suitability of irradiation for various forms of carrier materials

    Country

    Carrier

    Indonesia peat, filter mud and (soil + charcoal)
    Korea peat
    Malaysia Compost from empty fruit bunch of oil palm (EFB)
    Philippines soil and charcoal
    Thailand peat
    Vietnam peat

    Table 4. Proper dose for irradiation sterilization
    Country Carriers Optimum dose Remarks
    Indonesia Peat 35kGy 35 kGy had been evaluated by bf producer years ago and the dose had been used to sterilize bf carrier up to now
    Malaysia EFB compost > 30 kGy  
    Philippines Soil plus charcoal
    (3: 1)
    30 kGy Clay loam soil
    Thailand Peat 8 kGy 80 % reduced
    Vietnam Peat 25 to 30 kGy  
  2. Negative effects
    High cost for new equipment, transportation cost, needs expertise and regulation for radiation facilities.

3-2. Autoclaving

  1. Positive effects
    Cheaper and complete sterilization for small scale production, easy to operate, low maintenance cost and ease of accessibility
  2. Negative effects
    Time and labour-consuming on mass production, possibility of change in chemical and physical properties of carrier materials

4. Comments

Irradiation sterilization is a viable alternative method to autoclaving. Irradiation is suitable for mass sterilization. The irradiation method will be gradually accepted with increasing acceptance of quality control on biofertilizers. Detailed studies are needed to determine the effects of irradiation on the biotic and abiotic properties of carriers.


Attachment IV

V. Strategies for Practical Application of Biofertilizer
Carriers Sterilized by Irradiation


  1. Objectives

    To supply biofertilizers of high quality to farmers and to provide better options for mass sterilization of biofertilizer carriers.

  2. Obstacles and constrains
    1. The main constraints for some participating countries are the inaccessibility to irradiation facilities and inconvenience to biofertilizer producers/manufacturers which include high transportation cost, and where there is no commercial multi-purpose irradiation facility ( Thailand and Vietnam ).
    2. There is still limited information on irradiation sterilization for biofertilizer producers/manufacturers to embark on commercial scale biofertilizer production with sterilized carriers.
    3. There is still limited research on practical application of irradiation sterilized biofertilizer carriers.
  3. Strategies
    1. Preparation of comprehensive information including benefits on use of biofertilizers and of irradiation sterilization methods to be submitted to the policy makers for decision on national program on irradiation sterilization on biofertilizer carries by 60 Co gamma irradiation or Electron Beam irradiation facilities
    2. Increasing dissemination of information on biofertilizer production and application through brochures, manuals, DVD, TV, radio and websites in native languages to producers/manufacturers, consumers and policy makers
    3. Conducting practical demonstration by producers/manufacturers and extension agencies to farmers and the public.
    4. Providing examples from countries having successful programs on biofertilizer production and application.

Attachment V

FNCA EVALUATION REPORT


Project name:

Biofertilizer

Project leader (country) :

China , Indonesia , Japan , Korea , Malaysia , Philippines , Thailand and Vietnam

Date:

November 22, 2006

Object of project:

To improve and disseminate biofertilizer technology
  1. Major outcome of project

    1. Development of Biofertilizers
      1. Selection methods for effective microorganisms have been developed. Nitrogen fixation activity was evaluated by 15 N tracer technique, Acetylene reduction assay (ARA) and DNA-PCR technique. 15 N dilution and N-difference methods were used for field evaluation ( China , Indonesia , Japan , Malaysia , Philippines , Thailand and Vietnam ). P solubilizing/P uptake/P absorption activities were evaluated by 32 P tracer technique ( Indonesia , Malaysia , Thailand and Vietnam ), and also by HPLC and spectrophotometric methods ( Korea ).
      2. The suitable carriers for biofertilizer have been selected as follows;
        - Rhizobium : peat, soil, charcoal, chicken dung, filter mud
        - Non-symbiotic nitrogen fixing bacteria: peat, filter mud, chicken dung, compost (EFB), Clay loam soil+charcoal (3:1)
        - Mycorrhiza: soil, sand, rice straw, vermiculite
        - P and K solubilizing bacteria: vermiculite, organic manure, peat moss, compost (EFB)
      3. Effectiveness of the following biofertilizers was reported;
        China : Rhizobium, Associative N fixer, P and K solubilizer
        Japan : Rhizobium , Rhizobium + Associative N fixer, VAM
        Indonesia: Non-symbiotic N fixer, PGPR
        Korea : P solubilizer, PGPR
        Malaysia : VAM, Non-symbiotic N fixer, P solubilizer, PGPR
        Philippines : Associative N fixer
        Thailand : R hizobium, Rhizobium + VAM, Non-symbiotic N fixer+VAM, PGPR
        Vietnam : Mixing culture of Rhizobium, P solubilizer +PGPR
      4. 15 N dilution method has been applied to evaluate the potential of nitrogen fixing activity of symbiotic ( Indonesia , Japan , Thailand and Vietnam ) and non-symbiotic bacteria ( China , Indonesia , Malaysia , and Philippines ). It was found that nitrogen fixing bacteria can contribute about 15 - 75% of plant N.
    2. Irradiation sterilization method
      1. Sterilization of carrier by irradiation has been considered. The appropriate gamma-ray dose was reported to be ranging from 5 to 50 kGy, depending on the carrier materials ( China , Indonesia , Japan , Korea , Malaysia , Philippines , Thailand and Vietnam ).
      2. Effect of carrier sterilization by irradiation
        Compared with autoclave method, irradiation method ensures higher survival rate of biofertilizer inoculants in the carriers, resulting in higher crop yield and better crop quality, in some cases. However, there is still need for further studies on irradiation dosage for complete sterilization of carrier materials.
    3. Economic Evaluation
      1. Field demonstration has been conducted in each country. It was demonstrated that yield was increased by inoculating biofertilizer as shown in Appendix I. In the case of soybean production in Thailand , yield was increased by 10-122 %, depending on soil properties. In the Philippines , with Bio-N inoculation + 1/2N+P+K, yield was increased by 34% relative to fertilizer NPK application.
      2. The economic advantages of biofertilizer usage are as follows;
        1. Reduce the application of chemical fertilizers, and increase the crop yield and quality.
          Indonesia: Reduce the application of N fertilizer close to 50% and increase grain yield of corn by 11- 22% by associative N 2 fixing bacteria
          Korea : Net yield increased by 48.7% for lettuce by Bacillus sp.
          Malaysia : Contribution of 1/3 of N requirement of young oil palm by biofertilizer from associative N 2 fixing bacteria.
        2. Increase in the net income
          China : Net income increased by 9.2% for wheat and 2.1 % for rice compared to NPK application
          Indonesia : Net income increased by 16% by Azospirillum +1/2N+PK for corn compared to 1/2N+PK application
          Japan : Net income increased by 14% by Rhizobium and 35% by Rhizobium + Azospirillum for soybean compared to NPK application
          Philippines : Net income increased by 58% by Bio-N+PK 81% by Bio-N+1/2N+PK for corn compared to NPK application
          Thailand : Net income increased by 88% by Rhizobium and 128% by Rhizobium +PK for soybean compared to no fertilizer in unfertile soil
          Vietnam : Net income increased by 12.31-21.21% for peanut compared to non inoculation of Rhizobium
  2. Evaluation indicators for above outcome

    Socio-economic impact
    (Application or Achievement of project object) : 4 point
    Scientific impact (Basic technology or Activity): 4 point

  3. Project outlook

    □ Continuation ■ Change/Revision □ Termination

    Reason: In Asia , sustainable agriculture is essentially important to support increasing population. Agro-environment is drastically changed by climate and intensive agronomic practices from improper and injudicious use of chemical fertilizers and other agrochemicals, and beneficial microorganisms are needed for sustainable agriculture. Gamma or Ion Beam radiation methods are proposed to be utilized in the development of new functional microorganisms for sustainable agro-environment.

    Proposal: Development of new functional microorganisms for sustainable agriculture

    Definition of new "functional microorganisms": organisms that have improved symbiotic and non-symbiotic properties, tolerant to biotic and abiotic stresses, and having abilities to promote plant growth and control plant pathogens.

    Plan 1: Selection of functional microorganisms in agriculture

    • Mutation breeding of functional microorganisms by irradiation
      -- Symbiotic and non-symbiotic microorganisms such as Rhizobium , Azospirillum, mycorrhiza, PGPR, antagonistic microorganisms for plant diseases, etc.
    • The use of isotope technique to evaluate effectiveness of functional microorganisms

    Plan 2: Irradiation sterilization of carriers for microorganisms

    • Establishment of suitable sterilization methods for carriers
    • Practical application of carriers sterilized by gamma and electron beam irradiation
  4. Ripple effect to other fields or Benefits to end-users

    FNCA Biofertilizer Manual was published on the basis of the information and experience of the BF group activities from 2002 to 2005. This manual was distributed to researchers, academicians and biofertilizer producers. New publications including brochures are proposed to be printed in native languages for farmers and extension workers. Biofertilizer Newsletters were edited by project leaders and distributed to FNCA participating countries. The Manual and Newsletters can be easily accessed and downloaded via FNCA website.

  5. Opinion about the project (problems, ideas, remarks, proposal)

    The participants feel that the biofertilizer project has positive socio-economic and scientific impacts. However, there is a need for a revision or a change of the project, to include substantiation on the functionality of the microorganisms using isotope techniques and further application of radiation including ion beam and electron beam for the development of new functional microorganisms.


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