Dự án nông nghiệp " Replacing fertiliser N with rhizobial inoculants for legumes in Vietnam for greater farm profitability and environmental benefits " MS7 doc

1

Ministry of Agriculture & Rural Development

Collaboration for Agriculture & Rural
Development


013/06VIE
Replacing fertiliser N with rhizobial
inoculants for legumes in Vietnam for
greater farm profitability and
environmental benefits



MS7: Capacity Improvement Report







September 2009


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Table of Contents

1. Institute Information 3
2. Contact Officers 3
3. Project Abstract 4
4. Executive Summary 4
5. Technical (Capacity Improvement) Report 7
5.1. Inoculant production and training by Institutes 8
5.2. Private sector distribution and extension of inoculants 25
5.3. Private sector production of inoculants 25
5.4. Training and extension 26


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1. Institute Information
Project Name
Replacing fertiliser N with rhizobial inoculants for
legumes in Vietnam for greater farm profitability
and environmental benefits
Vietnamese Institution
Oil Plants Institute (OPI)
Vietnamese Project Team Leader
Ms Tran Yen Thao
Australian Organisation
NSW Industry & Investment
University of New England
University of Sydney
Australian Personnel
Dr David Herridge; Dr Roz Deaker
Ms Elizabeth Hartley, Mr Greg Gemell
Date commenced
March 2007

Completion date (original)
March 2009
Completion date (revised)
November 2009
Reporting period
December 2008 – September 2009

2. Contact Officer(s)
In Australia: Team Leader
Name:
Dr David Herridge
Telephone:
02 67631143
Position:
Professor, Soil
Productivity
Fax:
02 67631222
Organisa
tion
University of New
England - PIIC
Email:


In Australia: Administrative contact
Name:
Mr Graham Denney
Telephone:
02 63913219

Position:
Manager External Funding
Fax:
02 63913327
Organisation
Industry & Investment NSW
Email:
graham.denney@dpi.
nsw.gov.au

In Vietnam
Name:
Ms Tran Yen Thao
Telephone:
08 9143024 – 8297336
Position:
Researcher
Fax:
08 8243528
Organisation
Oil Plants Institute
(OPI)
Email:

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3. Project Abstract




















4. Executive Summary
Objective assessment of capacity and competency of trained staff to
produce high quality inoculants and to train farmers and extension
workers in successful application in the field
During 2007-08 the three collaborating institutes, OPI, IAS and SFI, produced
inoculants and had them tested for quality by OPI. Each institute used slightly
different production technologies. The number of samples tested was 261 from a total

of 465 samples received (60 from IAS, 180 from SFI and 225 from OPI). There were
36, 108 and 117 samples tested from IAS, SFI and OPI, respectively. 117 samples
were tested in 2007 and 144 samples in 2008.

Results indicated that the number of rhizobia in the inoculants ranged between <10
6
to
>10
9
cfu/g and the number of contamination ranged <10
5
to >10
6
cfu/g. However,
most inoculants (72%) had high rhizobial counts, between 5 x 10
8
and 3 x 10
9
cfu/g,
and were low in contamination (≤10
6
cfu/g). Plant-injection MNP counts were <10
5
to
>10
8
rhizobia/g moist peat but most inoculants (75%) had ≥10
8
rhizobia/g. Current
standards for peat inculants in Australia are ≥ 1x10

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rhizobial cfu/g moist peat and
contaminants are not detectable at a dilution level of 10
-6
; MPN plant-infection counts
are ≥10
8
. Therefore, in relative terms, a high percentage of legume inoculants now
produced by the three project institutes in Vietnam can meet the requirements for
production of high quality inoculants.
Farmers in Vietnam currently fertilise legumes such as soybean and groundnut with N,
rather than inoculate with rhizobia. Replacing fertiliser N with rhizobial inoculants
would save Vietnamese farmers A$50-60 million annually in input costs and, at the same
time, help facilitate the desired expansion in legume production. There would also be
positive environmental outcomes. This project aims to increase production of high-
quality legume inoculants in Vietnam through enhanced production capacity,
implementation of a national quality assurance (QA) program and increased inoculant
R&D. Participating in the project in Vietnam are the Oil Plants Institute (OPI), the
Institute of Agricultural Science (IAS) and the National Institute for Soils and Fertilisers
(NISF), now known as the Soils & Fertilisers Institute (SFI). Institutions in Australia are
NSW Department of Primary Industries and the University of Sydney. Legume inoculant
use by farmers in Vietnam will be increased through the development and
implementation of an effective extension and training program for researchers, MARD
extension officers and farmers. The benefits of inoculants and legume nitrogen fixation
will be demonstrated in the field and communicated through workshops, meetings and
publications. To ensure sustainability of inoculant production and use, the project will
engage the private sector in marketing and ‘pilot production’ of legume inoculants, with
the aim that they may scale-up production and progressively take over supply as the
technology and markets are developed.

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The variation in rhizobial and contaminant counts for the different batches resulted
from differences in procedures and expertise amongst the three laboratories. The
number of rhizobia/g inoculant was unstable between production batches in 2007 but
improved during 2008. This mainly resulted from improvement in moisture content of
inoculants at OPI and sterilization process at SFI. At IAS, the problems with
sterilization and moisture content were not properly resolved, resulting in less
consistency and lower quality of inoculants. However, IAS continues improving
carrier formulations that will improve low quality peat.

The training program for farmers and extension workers was based on their
involvement in field demonstrations, attendance in workshops, exposure to extension
literature and application of inoculants on their farms. The field demonstrations were
simple, multi-location inoculation experiments in the legume production areas of the
country.

During 2007–09, a total of 168 demonstration (extension) trials were conducted in 10
provinces. The demonstration fields usually had two treatments: +inoculation and –
inoculation (farmer
’
practice with N fertilisers). Farmers were invited to the
demonstration fields at least once. At many fields such as in DakNong and DakLak,
they also came to the fields 2–3 times at nodule and biomass harvest as well as grain
harvest time. At each trial site, at least 20 farmers, extension officers, agriculture
advisors came and made evaluation of the trial (3400+ person visits to the extension
trials). Researchers at the project institutes trained farmers and extension workers on
how inoculants work, how to apply inoculants to seed, how to determine if the
inoculated crops are fixing nitrogen well and how to record results. They observed

development of soybean and groundnut and compared the health and growth of the
plants in the inoculated and N-fertilised treatments. They dug plants from the soil to
observe nodules and learnt to recognise effective nodules with pink colour inside as
distinct from the white ineffective nodules. They learnt to evaluate inoculation
benefits by sampling soybean and groundnut plants, weighing biomass and grains.
Farmers were very interested in learning about nitrogen fixation and asked many
questions.

As well, 20 training workshops for farmers were conducted in 10 provinces. In each
workshop, 30–50 farmers, extension workers/officers and other (agricultural persons,
officers) participated, with around 800 persons in total at the workshops. Farmers and
extension workers who participated in the field experiments and demonstrations were
usually involved. At the workshop farmers were be supplied information on legume
nitrogen fixation and its benefits, and on inoculants and how they are used. Each of
the three project institute prepared their own presentations. Flyers (about 1500) were
prepared and handed out to the farmers.

Capacity of the private sector to provide high quality inoculant
distribution systems, together with sound advice to farmers in their use


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There is an increasing capacity of the private sector to distribute inoculants and
support farmers in their use. Three private-sector companies are now actively involved
in the project - Private Business Ngoc Trung at Son La Town, Son La province, Viet
A Nghia Dan Joint Stock Company at Nghia Dan, Nghe An province and Komix at
Binh Duong province. Cu Chi Bio-Chemical Fertiliser Joint Stock Company would
like to distribute inoculants for farmers but are having problems arranging staff. It is
hopeful that IAS can collaborate with Cu Chi company in the future for distribution
and marketing. Ngoc Trung and Viet A Nghia Dan are young companies but they are

interested in production of bio-products like rhizobial inoculants and have great
potential for production and distribution. With support from SFI about delivery of
broth cultures and technology transfer, the two companies produced and supplied
inoculants for 90 ha of groundnut and soybean, for Nghe An and Son La provinces.

Komix now has a network for distributing their bio-fertilizers covering almost half of
the country, from the center to the south. Komix conducted field trials with inoculants
produced by OPI. During 2008, they conducted one soybean field experiment at
DakNong province and completed 15 field demonstrations in DakNong and Tay Ninh
provinces. Komix supplied inoculants for several ha of soybean and groundnut where
they conducted field trials.
Assessment of the improved capacity of private sector companies in the
production of high quality inoculant
The potential market for inoculants in Vietnam could be around 500 tonnes annually,
assuming application rates of 1 kg inoculant/ha and about 50% of legume area
inoculated. This amount substantially exceeds the capacity of the project institutes for
production (currently <20 tonnes annually). Therefore, it was considered necessary to
involve the private sector in commercial production.

Inoculant production at Son La and Nghe An companies – SFI produces the broth
cultures and supplies them to the companies. The broth cultures are injected into
sterilized peat prepared by the companies. SFI has transferred the technologies of peat
sterilization, injection, packaging and storage. The quality of inoculants has been
monitored by SFI throughout the production process. Final products were tested by an
independent quality control laboratory. Some pilot batch production was done and
inoculants were supplied for 90 ha of groundnut and soybean in Son La and Nghe An.
However, the cost of production was found to be high due to high cost of
transportation. Nghe An and Son La are around 300 km far from Hanoi where the
broths were produced by SFI. In addition, there is always the risk that the broths might
be damaged during transport from Hanoi to Son La.

The Komix company works in the project with OPI. There are 2 steps of the process.
In the first step they field-test inoculants produced by the institute, and market and
distribute them to farmers. Komix gets feedback of the potential of the inoculant based
on evaluation of biological and economic benefits of inoculation in the target
provinces. OPI transferred technologies to Komix including field experiments/trials


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and extension documents, extension activities (field days at farms, farmer workshops)
and together with Komix took care of field experiments and demonstrations. The
second step is to set up production at Komix. It is proposed for medium scale
production at Komix and then eventually increased production. OPI manages quality
control of inoculants produced by Komix, supplies mother culture annually, improves
production technology and promotes the products. OPI is arranging for a pilot
production at Komix in December 2009. Inoculants will be supplied on order to Dong
Thap province for soybean and groundnut (100 ha soybean and 20 ha groundnut).
Copies of training and extension materials including training reports and
lessons learned from study tours
Copies of training and extension materials are attached as appendices. In terms of
training of institute personnel in inoculant technology, there were four major events:
• Quality assurance workshop, IAS, HCM City Feb-March 2007. There were 17
participants drawn from collaborating institutes and private sector companies.
The workshop was designed by ALIRU staff and presented by Elizabeth
Hartley (ALIRU), Greg Gemell (ALIRU) and Rosalind Deaker (University of
Sydney). Each participant was provided with a workbook and comprehensive
notes on procedures:
Workbook: Quality Control of Legume Inoculants Workshop, 26
th
February –

9
th
March 2007, compiled by E. Hartley, G. Gemell, J. Hartley (01/02/2007).
37 pages.
Procedures Section: Quality Control of Legume Inoculants Workshop,
compiled by E. Hartley, G. Gemell, J. Hartley (01/02/2007). 32 pages.
• Inoculant technologies training workshop, Suranaree University of
Technology, Thailand June 2007. Three project scientists participated.
Activities focussed on lectures, laboratory training and visits to production
facilities in Thailand. The training covered background information on
rhizobia, fermentation technologies including large systems and the micro
production unit (MPU), carbon source for rhizobia in fermentation, other plant
growth promoting (PGP) organisms and counting techniques.
• Inoculant training workshop, Suranaree University of Technology, Thailand
October 2008. Four project scientists participated. Activities focussed on
practice rather than theory and consisted of recognition of Rhizobium strains,
small- and large-scale inoculant production and quality control. Participants
from each institute concentrated on particular aspects that were particularly
relevant to them.
• Study tour, Yen Thao to Australia November-December 2008. Activities
focused on research on identification of rhizobial strains, survival of the
Australian, Vietnamese strains as well as strains from other countries;
participated in the review of quality control and QA system currently in


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Australia and prepared documents for protocols of inoculant production and
quality control of legume inoculants in order to apply in Vietnam.
The research was conducted at the Department of Agriculture, Food and Natural
resources, the University of Sydney, from 20 to 28 November and from 15 to 20

November, at the Australian Legume Inoculant Research Unit, Department of
Primary Industries, NSW, Gosford.
5. Technical Report
The technical report addresses the required reporting headings.
1. Objective assessment of capacity and competency of trained staff to produce
high quality inoculants and to train farmers and extension workers in
successful application in the field
2. Capacity of the private sector to provide high quality inoculant distribution
systems, together with sound advice to farmers in their use
3. Assessment of the improved capacity of private sector companies in the
production of high quality inoculant
4. Copies of training and extension materials including training reports and
lessons learned from study tours
5.1. Objective assessment of capacity and competency of trained staff to
produce high quality inoculants and to train farmers and extension
workers in successful application in the field
5.1.1. Testing inoculants produced by the three project institutes

All participating institutes produced inoculants using their own technologies and sent
samples to OPI for quality testing. We followed standardized methods of testing of
ALIRU (Australia) with some change to suit laboratory conditions in Vietnam. During
the two years 2007–2008, the number of samples tested was 261 from a total of 465
samples received (60 from IAS, 180 from SFI and 225 from OPI). There were 36, 108
and 117 samples tested from IAS, SFI and OPI, respectively. 117 samples were tested
in 2007 and 144 samples in 2008.

The information on tested samples is shown in the Table 1. The information was
source of inoculants, batch number of production, types of inoculant (formulation),
legume hosts, rhizobial strain, date of arrival, date of the batch testing and lab code for
the samples. Products were mainly peat-based inoculants for groundnut and soybean.

Some liquid inoculants were submitted by OPI (OPI-L-S-B3-CB1809, OPI-L-S-B3-
SL1, OPI-L-S-B3-SL2). NC 92, CB 1809 and 4 local strains (GL1, GL2 for
groundnut and SL1, SL2 for soybean) were used in the inoculants.



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Moisture content is showed in the Table 2. Generally moisture content of inoculants
produced in Vietnam varied in the range 20–52%, because of different qualities of the
carriers and production technologies applied in the institutes. However, moisture
content of most inoculants was 38–49% (80% samples) (Graph 1). For peat
inoculants, around 40–50% moisture content is optimum.

At IAS moisture content of inoculants fluctuated from 20 to 52% (Graph 2) while it
was more stable at ISF and OPI, ranged 38–46% (79% samples).

Moisture content of inoculants tested dur ing 2007-2008
0
10
20
30
40
50
60
051015
Production batch
Moi sture content (%)
QA Moisture content Year
2007
QA Moisture content Year

2008





Graph 2. Moisture content by institute
s
0
10
20
30
40
50
60
0246810
Batch production
Miosture content (%)
IAS
SFI
OPI



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Numbers of rhizobia in the inoculants were enumerated using direct plate counting
and plant-infection, most probable number (MPN) counting. With direct plate
counting, rhizobial numbers ranged from <10
6

to >10
9
(Table 3). The plant-infection
MPN counts closely matched those of the plate counts and ranged from <10
5
to >10
8

(Table 4). Most inoculant samples tested (71%) had rhizobial counts between 10
8
to
10
9
cells/g, a very good result. Twenty six percent of samples tested had counts in the
range 10
5
to 10
7
. There were few inoculants in which rhizobial cells did not exist or
could not be detected because contaminants (mainly fungal) had overun the rhizobia.

The necessity of MPN counting was highlighted with IAS Batch 3 and SFI Batch 1
when plate counting was impossible because of the large background populations of
contaminating organisms (Table 5).

Numbers of contaminants in the inoculants are shown in Table 5. Almost all samples
had number of contaminants from 10
5
to 10
6

cells/g peat (75%). Some samples had
very low contaminant counts, i.e. <10 cells/g, while few other samples had high levels
of contaminants, around 10
7
cells/g.

Table 1: General information of inoculants tested

IAS-produced inoculants
Year 2007
Batch number 1 2 3
Type of inoculants Peat Peat Peat
Legume host Groundnut
Soybean
Groundnut
Soybean
Groundnut
Soybean
Rhizobium strain
name
NC 92
CB 1809
NC 92
CB 1809
NC 92
CB 1809
Date of arrival 4/7/2007 16/7/2007 16/8/2007
Date the batch tested 6/7/2007 17/7/2007 17/8/2007
Lab code of samples IAS-P-B1-G-NC92 (1-5)
IAS-P-B1-S-BC1809 (1-5)

IAS-P-B2-G-NC92 (1-5)
IAS-P-B2-S-BC1809 (1-5)
IAS-P-B3-G-NC92 (1-5)
IAS-P-B3-S-BC1809 (1-5)
Year 2008

Batch number 1 2 3
Type of inoculants Peat Peat -
Legume host Groundnut Groundnut -
Rhizobium strain
name
NC 92
GL1
GL2
NC 92
GL1
GL2
-
Date of arrival 15/5/2008 10/9/2008 -
Date the batch tested 15/5/2008 12/9/2008 -
Lab code of samples IAS-P-B1-G-NC92 (1-5)
IAS-P-B1-G-GL1 (1-5)
IAS-P-B1-G-GL2 (1-5)

IAS-P-B2-G-NC92 (1-5)
IAS-P-B2-G-GL1 (1-5)
IAS-P-B2-G-GL2 (1-5)
-

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SFI-produced inoculants
Year 2007
Batch number 1 2 3
Type of inoculants Peat Peat Peat
Legume host Groundnut
Soybean
Groundnut
Soybean
Groundnut
Soybean
Rhizobium strain
name
NC 92
GL1
GL2
CB 1809
SL1
SL2
NC 92
GL1
GL2
CB 1809
SL1
SL2
NC 92
GL1
GL2
CB 1809
SL1

SL2
Date of arrival 9/5/2007 10/10/007 20/11/2007
Date the batch tested 10/5/2007 12/10/2007 21/11/2007
Lab code of samples ISF-B1-NC92 (1-5)
ISF-B1-SL1 (1-5)
ISF-B1-GL2 (1-5)
ISF-B1-CB1809 (1-5)
ISF-B1-SL1 (1-5)
ISF-B1-SL2 (1-5)

ISF-B2 NC92 (1-5)
ISF-B2 SL1 (1-5)
ISF-B2 GL2 (1-5)
ISF-B2-CB1809 (1-5)
ISF-B2-SL1 (1-5)
ISF-B2-SL2 (1-5)

ISF-B3—NC92 (1-5)
ISF-B3 SL1 (1-5)
ISF-B3-GL2 (1-5)
ISF-B3-CB1809 (1-5)
ISF-B3-SL1 (1-5)
ISF-B3-SL2 (1-5)

Year 2008

Batch number 1 2 3
Type of inoculants Peat Peat Peat
Legume host Groundnut
Soybean

Groundnut
Soybean
Groundnut
Soybean
Rhizobium strain
name
NC 92
GL1
GL2
CB 1809
SL1
SL2
NC 92
GL1
GL2
CB 1809
SL1
SL2
NC 92
GL1
GL2
CB 1809
SL1
SL2
Date of arrival 22/5/2008 20/8/008 1/9/2008
Date the batch tested 23/5/2008 22/8/2008 4/9/2008
Lab code of samples ISF-B1-G-NC92 (1-5)
ISF-B1-G-SL1 (1-5)
ISF-B1-G-GL2 (1-5)
ISF-B1-CB1809 (1-5)

ISF-B1-SL1 (1-5)
ISF-B1-SL2 (1-5)

ISF-B2-G-NC92 (1-5)
ISF-B2-G-SL1 (1-5)
ISF-B2-G-GL2 (1-5)
ISF-B2-CB1809 (1-5)
ISF-B2-SL1 (1-5)
ISF-B2-SL2 (1-5)

ISF-B3-G-NC92 (1-5)
ISF-B3-G-SL1 (1-5)
ISF-B3-G-GL2 (1-5)
ISF-B3-CB1809 (1-5)
ISF-B3-SL1 (1-5)
ISF-B3-SL2 (1-5)

IOOP-produced inoculants
Year 2007
Batch number 1 2 3
Type of inoculants Peat Peat Liquid
Legume host Groundnut
Soybean
Groundnut
Soybean
Soybean


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Rhizobium strain

name
NC 92
GL1
GL2
CB 1809
SL1
SL2
NC 92
GL1
GL2
CB 1809
SL1
SL2
-
-
-
CB 1809
SL1
SL2
Date of arrival 2/4/2007 20/10/2007 22/12/2007
Date the batch tested 4/4/2007 25/10/2007 25/12/2007
Lab code of samples OPI-G-B1-NC92 (1-5)
OPI-G-B1-SL1 (1-5)
OPI-G-B1-GL2 (1-5)
OPI-S-B1-CB1809 (1-5)
OPI-S-B1-SL1 (1-5)
OPI-S-B1-SL2 (1-5)

OPI-G-B2-NC92 (1-5)
OPI-G-B2-SL1 (1-5)

OPI-G-B2-GL2 (1-5)
OPI-S-B2-CB1809 (1-5)
OPI-S-B2-SL1 (1-5)
OPI-S-B2-SL2 (1-5)

-
-
-
OPI-L-S-B3-CB1809 (1-5)
OPI-L-S-B3-SL1 (1-5)
OPI-L-S-B3-SL2 (1-5)

Year 2008

Batch number 1 2 3
Type of inoculants Peat Peat Peat
Legume host Groundnut
Soybean
Groundnut
Soybean
Groundnut
Soybean
Rhizobium strain
name
NC 92
GL1
GL2
CB 1809
SL1
SL2

NC 92
GL1
GL2
CB 1809
SL1
SL2
NC 92
GL1
GL2
CB 1809
SL1
SL2
Date of arrival 1/3/2008 6/4/2008 1/7/2008
Date the batch tested 1/3/2008 6/4/2008 1/7/2008
Lab code of samples OPI-G-B1-NC92 (1-5)
OPI-G-B1-SL1 (1-5)
OPI-G-B1-GL2 (1-5)
OPI-S-B1-CB1809 (1-5)
OPI-S-B1-SL1 (1-5)
OPI-S-B1-SL2 (1-5)

OPI-G-B2-NC92 (1-5)
OPI-G-B2-SL1 (1-5)
OPI-G-B2-GL2 (1-5)
OPI-S-B2-CB1809 (1-5)
OPI-S-B2-SL1 (1-5)
OPI-S-B2-SL2 (1-5)

OPI-G-B3-NC92 (1-5)
OPI-G-B3-SL1 (1-5)

OPI-G-B3-GL2 (1-5)
OPI-L-S-B3-CB1809 (1-5)
OPI-L-S-B3-SL1 (1-5)
OPI-L-S-B3-SL2 (1-5)

Batch number 4 5 6
Type of inoculants Peat Peat Peat
Legume host Groundnut
Soybean
Groundnut
Soybean
Groundnut
Soybean
Rhizobium strain
name
NC 92
CB 1809

NC 92
CB 1809

NC 92
CB 1809

Date of arrival 2/8/2008 5/10/2008 10/11/2008
Date the batch tested 2/8/2008 5/10/2008 10/11/2008
Lab code of samples OPI-B4-NC92 (1-10)
OPI-B4-CB1809(1-10)

OPI-B5-NC92 (1-10)

OPI-B5-CB1809 (1-10)

OPI-B6-NC92 (1-10)
OPI-B6-CB1809 (1-10)




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Table 2: Moisture content of inoculants

Moisture content (%) Inoculants
Batch 1 Batch 2 Batch 3
IAS
Year 2007
IAS-G-NC92
IAS-S-CB1809
Average
44.4
42.2
43.3
54.6
49.5
52.1
-
-
-
IAS
Year 2008

Batch 3 Batch 4
IAS-G-NC92
IAS-G-GL1
IAS-G-GL2
Average
38.8
39.5
40.0
39.4
19.8
20.4
20.6
20.3
-
-
-


SFI
Year 2007
Batch 1 Batch 2 Bathch 3
ISF-G-NC92
ISF-G-SL1
ISF-G-GL2
ISF-CB1809
ISF-SL1
ISF-SL2
Average
42.3
42.3

41.2
42.1
42.8
41.5
42
34.9
38.6
34.7
37.0
31.8
34.5
35.2
45.6
43.2
44.8
46.1
43.7
42.5
44.3
Year 2008
Batch 4 Batch 5 Bathch 6
ISF-G-NC92
ISF-G-GL1
ISF-G-GL2
ISF-CB1809
ISF-SL1
ISF-SL2
Average
36.5
37.0

37.6
36.8
32.5
35.5
36.0
41.3
31.9
30.2
41.6
38.2
45.3
38.9
41.3
31.9
30.2
41.6
38.2
45.3
38.1
IOOP

Year 2007
Batch 2 Batch 2 Batch 3
OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
OPI-S-CB1809
OPI-S-SL1
OPI-S SL2
Average

20.0
21.2
20.7
20.5
21.3
20.6
20.7
45.5
46.6
45.8
44.7
44.6
45.8
45.5
-
-
-
-
-
-

Year 2008
Batch 4 Batch 5 Batch 6
OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
OPI-S-CB1809
36.7
36.0
36.8

38.2
37.3
38.2
37.9
38.4
41.1
37.5
36.4
38.6


14
OPI-S-SL1
OPI-S SL2
Average
37.6
36.9
37.0
40.0
39.3
38.5

39.2
37.3
38.4
Batch7 Batch 8 Batch 9
OPI-G-NC92
OPI-G-NC92
OPI-G-NC92
OPI-S-CB1809

OPI-S-CB1809
OPI-S—CB1809
Average
36.2
36.3
40.1
37.5
38.6
37.7
37.7
38.3
37.4
38.2
34.8
39.4
40.2
38.2
39.2
37.4
40.5
38.7
39.6
40.7
39.4



15

Table 3: Number of viable rhizobia by direct plate count

Number/g Samples
Batch 1 Batch 2 Batch 3
IAS

Year 2007

IAS-NC92
1
2
3
IAS-BC1809
1
2
3
Average


68 x 10
6

27 x 10
6
51 x 10
6

29 x 10
6

53 x 10

6

12 x 10
6

37x10
6


9 x 10
7

16 x 10
7
17 x 10
7
63 x 10
7

0.3 x 10
7

43 x 10
7

27x10
7


Fungi cont

*
Fungi cont
*
20 x 10
5


6 x 10
5

Fungi cont
*
Fungi cont
*
14x10
5

Year 2008

IAS-NC92
IAS-GL1
IAS-GL2
Average
53x10
6

16x10
6

33x10

6

34x10
6
0
0
0
-
-
-
SFI


Year 2007

ISF-NC92
ISF-SL1
ISF-GL2
ISF-CB1809
ISF-SL1
ISF-SL2
Average
Fungi cont.
Fungi cont.
Fungi cont.
Fungi cont.
Fungi cont.
Fungi cont.

33 x 10

6
20 x 10
6
25 x 10
6
36 x 10
6

7 x 10
6
16 x 10
6

29x10
6

55 x 10
8

45 x 10
8
12 x 10
8

24 x 10
8

19 x 10
8

16 x 10
8

28X10
8

Year 2008

ISF-NC92
ISF-GL1
ISF-GL2
ISF-CB1809
ISF-SL1
ISF-SL2
Average

23x10
7

32x10
7

14x10
8

68x10
7

20x10
7

12x10
7

49x10
7

22x10
8

34x10
8

12x10
8

18x10
8

12x10
8

87x10
7

18x10
8

10x10
8

40x10
8

52x10
8

90x10
7

47x10
7

64x10
7

20x10
8
IOOP
Bacth1 Bacth2 Bacth3
Year 2007

OPI-NC92
OPI-SL1
20 x 10
5

45 x 10
5

-
-
-
-


16
OPI-GL2
OPI-CB1809
OPI-SL1
OPI-SL2
Average
25 x 10
5

60 x 10
5

10 x 10
5

22 x 10
5

30x10
5

-
50 x 10
8

43 x 10
8

6 x 10
8

33x10
8
-
40 x 10
8

20 x 10
8

25 x 10
8

28x10
8

Year 2008
Bacth1 Bacth2 Bacth3
OPI-NC92
OPI-SL1
OPI-GL2
OPI-CB1809
OPI-SL1
OPI-SL2

Average
40x10
8

22x10
8

12x10
8

21x10
8

20x10
8

70x10
7

20x10
8
22x10
8

67x10
7

82x10
7

13x10
8

10x10
8

8x10
8

11x10
8
12x10
8

21x10
8

43x10
8

11x10
8

25x10
8

32x10
8

24x10

8
Batch 4 Batch 5 Batch 6
OPI-NC92
OPI-NC92
OPI-NC92
OPI-CB1809
OPI-CB1809
OPI-CB1809
Average

52x10
7

46x10
7

13x10
8

57x10
7

20x10
8

12x10
8

10x10
8

12x10
8

23x10
8

10x10
8

18x10
8

12x10
8

76x10
7

14x10
8

23x10
8

11x10
8

48x10
8

75x10
7

21x10
8

16x10
8

21x10
8



17

Table 4: Number of viable rhizobia by plant-infection MPN

Number/g Samples
Batch 1 Batch 2 Batch 3
IAS
Year 2007
IAS-P-G-NC92
1
2
3
IAS-P-S-CB1809
1
2

3
Average

84 x 10
6

26 x 10
6

44 x 10
6

132 x 10
6

57 x 10
6

42 x 10
6

64x10
6


97 x 10
7

45 x 10
7

96 x 10
7


85 x 10
7

42 x 10
7

59 x 10
7



58 x 10
6

42 x 10
6
25 x 10
6

12 x 10
5
59 x 10
5
30 x 10
6

Year 2008 Batch 4 Batch 5
IAS-NC92
IAS-GL1
IAS-GL2

Undetected
*
Undetected
*
Undetected
*
No rhizobia
No rhizobia
No rhizobia

-
-
-
ISF

Year 2007

ISF-G-NC92
ISF-G-SL1
ISF-G-GL2
ISF-CB1809
ISF-SL1
ISF-SL2

No rhizobia

No rhizobia
No rhizobia
No rhizobia
No rhizobia
No rhizobia
8 x 10
5
22 x 10
5

47 x 10
5

75 x 10
5

59 x 10
5

57 x 10
5
91 x 10
8

42 x 10
8

35 x 10
8

84 x 10
8

47 x 10
8

59 x 10
8
Year 2008

ISF-G-NC92
ISF-G-SL1
ISF-G-GL2
ISF-CB1809
ISF-SL1
ISF-SL2

13x10
8

28x10
7
11x10
7
10x10
7

30x10
6
70x10

6

30x10
7

14x10
8

22x10
8
30x10
7

22x10
8
60x10
7

42x10
7

34x10
8
23x10
7
60x10
7

50x10
7

70x10
7

IOOP

Year 2007

OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
OPI-S-CB1809
42 x 10
5

14 x 10
5

9 x 10
5

9 x 10
5
-
-
-
42 x 10
8

-
-

-
-


18
OPI-S-SL1
OPI-S SL2

68 x 10
5
16 x 10
5


36 x 10
8

22 x 10
8

-
-
Year 2008
Batch 1 Batch 2 Bacth 3
OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
OPI-S-CB1809
OPI-S-SL1
OPI-S SL2

16x10
7

17x10
7

27x10
7

13x10
7

20x10
7

80x10
6
70x10
7

17x10
7

58x10
7

26x10
8

15x10
8

80x10
7
11x10
8

7x10
8

24x10
8

23x10
8

10x10
8

32x10
8

Year 2008
Batch 4 Batch 5 Batch 6
OPI-G-NC92
OPI-G-NC92
OPI-G-NC92
OPI-S-CB1809
OPI-S-CB1809

OPI-S-CB1809

14x10
7

36x10
7

10x10
8

50x10
7

30x10
7

80x10
7
30x10
7

90x10
7

50x10
7

10x10
8

26x10
7

50x10
7
17x10
8

90x10
7

40x10
7

33x10
7

50x10
7

40x10
7


*¨
undetected: rhizobia could not be detected as samples were contaminated by a fungus that
damaged groundnut root (turned to black color).

19
Table 5: Number of contaminants in inoculants

Number/g Samples
Batch 1 Batch 2 Batch 3
IAS

Year 2007

IAS-P-G-NC92
1
2
3
IAS-P-S-CB1809
1
2
3
Average

28 x 10
5

76 x 10
5

90 x 10
5


16 x 10

5

74 x 10
5

43 x 10
5

55x10
5


13 x 10
4

82 x 10
4

32 x 10
4


17 x 10
4
49 x 10
4

91 x 10
4

47x10
4


14 x 10
6

32 x 10
6
24 x 10
6


54 x 10
6
39 x 10
6
11 x 10
6

29x10
6
Year 2008

IAS-NC92
IAS-GL1
IAS-GL2
Average
48x10
5

14x10
6

33x10
5

32x10
5
<10
<10
<10

-
-
-
ISF Batch 1 Batch 2 Batch 3
Year 2007

ISF-G-NC92
ISF-G-GL1
ISF-G-GL2
ISF-CB1809
ISF-SL1
ISF-SL2
Average

Fungi cont.
Fungi cont.
Fungi cont.

Fungi cont.
Fungi cont.
Fungi cont.

50 x 10
5

23 x 10
5

40 x 10
5

15 x 10
5
60 x 10
4
50 x 10
4

23x10
5

25 x 10
3

48 x 10
3

87 x 10

3

56 x 10
3

15 x 10
3

36 x 10
3

45x10
3

Year 2008

ISF-G-NC92
ISF-G-GL1
ISF-G-GL2
ISF-CB1809
ISF-SL1
ISF-SL2
Average

40 x 10
4

29 x 10
5

44x 10
5

20 x 10
6
90 x 10
5
45 x 10
5

69x10
5

18 x 10
5

45 x 10
5

88x 10
4

11 x 10
5
30 x 10
5
36 x 10
5

25x10

5


<100
<100
<100
<100
<100
<100

IOOP

Year 2007 Batch 1 Batch 2 Batch 3
OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
20 x 10
3

36 x 10
3

27 x 10
3

-
-
-
-
-

-


20
OPI-S-CB1809
OPI-S-SL1
OPI-S SL2
Average

54 x 10
3

42 x 10
3

32 x 10
3
35x10
3
25 x 10
3

72 x 10
3

37 x 10
3

45x10
3

<100
<100
<100
Year 2008 Batch 1 Batch 2 Batch 3
OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
OPI-S-CB1809
OPI-S-SL1
OPI-S SL2
Average

62 x 10
4

23 x 10
4

14 x 10
4

12 x 10
5
36 x 10
4
13 x 10
5

64x10
4

21 x 10
5

10 x 10
6

21 x 10
5

11 x 10
5
52 x 10
4
36 x 10
5

32x10
5

12 x 10
5

65 x 10
4

70 x 10
4

18 x 10

5
17 x 10
5
24 x 10
5

14x10
5

Year 2008 Batch 4 Batch 5 Batch 6
OPI-G-NC92
OPI-G-SL1
OPI-G-GL2
OPI-S-CB1809
OPI-S-SL1
OPI-S SL2
Average
22 x 10
5

23 x 10
5

16 x 10
5

10 x 10
5
40 x 10
4

13 x 10
5

15x10
5

32 x 10
3

14 x 10
3

10 x 10
4

52 x 10
3
34 x 10
3
38 x 10
3

45x10
3

<100
<100
<100
<100
<100

<100




21

5.1.2. Institute capacity to produce high quality of inoculants

The results of QA on 261 inoculants produced by the three collaborating Vietnamese
institutes during 2007-2008 above showed that the number of rhizobia ranged between
<10
6
to >10
9
cfu/g inoculant and number of contamination was <10
5
to >10
6
cfu/g.
However, the number of rhizobia/g was high in most inoculants (72% ), between 5 x
10
8
and 3 x 10
9
cfu/g, and low in contamination (≤10
6
cfu/g). Plant-injection MNP
counts were <10
5

to >10
8
rhizobia/g moist peat but most inoculants (75%) had ≥10
8

rhizobia/g. Current standards for peat inculants in Australia are ≥ 1x10
9
rhizobial cfu/g
moist peat and contaminants are not detectable at a dilution level of 10
-6
; MPN plant-
infection counts are ≥10
8
. Therefore, in relative terms, a high percentage of legume
inoculants now produced in Vietnam can meet the requirements for production of high
quality inoculants.

The variation in rhizobial and contaminant counts for the different batches resulted
from differences in procedures and expertise amongst the three laboratories. The
number of rhizobia/g inoculant was unstable between production batches in 2007
(Graph 3) but was stable during 2008. This resulted from improvement of production
technology. In the second year of the project the quality of inoculants produced at SFI
and OPI was much improved, 10
8
–10
9
rhizobia/g and contaminant <10
7
cells/g and no
batches were over-run with contaminants (Graphs 3 and 4, Tables 3–5). This mainly

resulted from improvement in moisture content of inoculants at OPI and sterilization
process at SFI. At IAS, the problems with sterilization and moisture content were not
properly resolved, resulting in less consistency and lower quality of inoculants.
However, IAS continues improving carrier formulations that will improve low quality
of peat.

Graph 3. Number of Rhizobium/g inoculant
0
2
4
6
8
10
02468101214
Production batch
Number of Rhizobium/g
inoculant (log)
Plate Count Year 2007
Plate Count Year 2008




22
Graph 4. Number of Rhizobium/g inoculant by institute
0
2
4
6
8

10
0246810
Production batch
Number of Rhizobium/g
inoculnat (log)
IAS
SFI
OPI


The quality control process at OPI worked extremely well and, clearly, the foundation
has been laid for a formal and expanded QA program in Vietnam in line with the
proposed expansion of inoculant production.

Graph 5. Number of contaminants in inoculant
s
0
1
2
3
4
5
6
7
8
0 5 10 15
Production batch
Numberof contaminants/g
inoculants (log)
Contami nants Year 2007

Contami nants Year 2008



5.1.3. Capacity of institute staff to train farmers and extension workers to
use inoculants

The training program for farmers and extension workers was based on their
involvement in field demonstrations, attendance in workshops, exposure to extension
literature and application of inoculants on their farms. The field demonstrations were


23
simple, multi-location inoculation experiments in the legume production areas of the
country. The experiments involve participation of farmers and extension officers in all
aspects, from the design of experiments to sowing, sampling, harvesting and
interpretation of results. The MARD extension service played a large role in extension
activities. Data from field demonstrations has used to compare the economics of
inoculant use versus the current practice of fertilizer N application. In the future, after
the project has officially ended, additional training courses will be organised for
farmers, extension workers and researchers in methods of inoculant use, and economic
as well as environmental benefits of inoculation.

During 2007–09, a total of 168 demonstration (extension) trials were conducted in 10
provinces. The demonstration fields usually had two treatments: +inoculation and –
inoculation (farmer
’
practice with N fertilisers). Farmers were invited to the
demonstration fields at least once. At many fields such as in DakNong and DakLak,
they also came to the fields 2–3 times at nodule and biomass harvest as well as grain

harvest time. At each trial site, at least 20 farmers, extension officers, agriculture
advisors came and made evaluation of the trial (3400+ person visits to the extension
trials). Researchers at the project institutes trained farmers and extension workers on
how inoculants work, how to apply inoculants to seed, how to determine if the
inoculated crops are fixing nitrogen well and how to record results. They observed
development of soybean and groundnut and compared the health and growth of the
plants in the inoculated and N-fertilised treatments. They dug plants from the soil to
observe nodules and learnt to recognise effective nodules with pink colour inside as
distinct from the white ineffective nodules. They learnt to evaluate inoculation
benefits by sampling soybean and groundnut plants, weighing biomass and grains.
Farmers were very interested in learning about nitrogen fixation and asked questions,
such as:

- How does the inoculant price?
- How much do inoculants used for 1000m
2
or 1 ha?
- Where can we purchase inoculants?
- Do inoculants have other benefits besides urea (N) fertiliser?
- Can we use inoculants together with plant protection products?
- Can we use legume inoculants for other crops?
- Are inoculants effected by bad weather such as heavy rain, hot weather?
- Can we use inoculants together with urea (fertiliser N)?

And main requests:

- Supply inoculants to farmers to test inoculants in their fields
- Technical support for farmers to use inoculants.

Farmers were given flyers introducing biological nitrogen fixation, benefits in terms

of chemical nitrogen fertiliser replacement, yield improvement and how to use
inoculants for their fields.



24
All participating institutes were successful with extension activities. SFI and IAS
staff organized field trials, invited farmers and extension workers to the fields and
trained them. OPI utilized extension officers more for the extension work with the
extension workers playing a principal role in the field trials. They prepared the trials
from the beginning to the end and liaised with the farmers while OPI staffs gave them
field trial procedures after discussions with them and extension documents. The OPI
staff and extension officers came together at the field demonstration sites at important
times (sowing, nodulation sampling, biomass and harvest sampling, field days) and
kept close contact during the course of the trials. The main objective of the
collaboration was that extension workers would become familiar in field trials and
demonstrations of legume inoculation and will be the key persons to train others
when inoculant use is spreading. As they are local extension persons they know very
well the current agricultural packages for peanut or/and soybean in an area, i.e. best
varieties, the package for plant protection, irrigation, fertiliser applications etc. This is
the case in Binh Dinh province (central coast), Dong Thap, An Giang and Tra Vinh
provinces (Mekong Delta).

In regarding to extension activities, 20 training workshops for farmers were conducted
in 10 provinces. In each workshop, 30–50 farmers, extension workers/officers and
other (agricultural persons, officers) participated, with around 800 persons in total at
the workshops. Farmers and extension workers who participated in the field
experiments and demonstrations were usually involved. At the workshop farmers were
be supplied information on legume nitrogen fixation and its benefits, and on
inoculants and how they are used. Each of the three project institute prepared their

own presentations. Flyers (about 1500) were prepared and handed out to the farmers.

5.2. Capacity of the private sector to provide high quality inoculant
distribution systems, together with sound advice to farmers in their use

At this stage three companies are actively involved in the project - Private Business
Ngoc Trung at Son La Town, Son La province, Viet A Nghia Dan Joint Stock
Company at Nghia Dan, Nghe An province and Komix at Binh Duong province. Cu
Chi Bio-Chemical Fertiliser Joint Stock Company would like to distribute inoculants
for farmers but are having problems arranging staff. It is hopeful that IAS can
collaborate with Cu Chi company in the future for distribution and marketing. Ngoc
Trung and Viet A Nghia Dan are young companies but they are interested in
production of bio-products like rhizobial inoculants and have great potential for
production and distribution. With support from SFI about delivery of broth cultures
and technology transfer, the two companies produced and supplied inoculants for 90
ha of groundnut and soybean, for Nghe An and Son La provinces.

Komix now has a network for distributing their bio-fertilizers covering almost half of
the country, from the center to the south (Appendix 5, in the second project report).
Komix conducted field trials with inoculants produced by OPI. They conducted one
soybean field experiment at DakNong province to see the effectiveness of nitrogen


25
fixation in terms of nodulation, biomass and seed yields and they completed 15 field
demonstrations in DakNong and Tay Ninh provinces. The data from the trials were
yield, income from inoculation and willingless to use inoculants by farmers. Komix
supplied inoculants for several ha of soybean and groundnut where they conducted
field trials.

5.3. Assessment of the improved capacity of private sector companies in
the production of high quality inoculant
The potential market for inoculants in Vietnam could be around 500 tonnes annually,
assuming application rates of 1 kg inoculant/ha and about 50% of legume area
inoculated. This amount substantially exceeds the capacity of the project institutes for
production (currently <20 tonnes annually). Therefore, it was considered necessary to
involve the private sector in commercial production.

In Vietnam there is no history of private companies producing substantial amounts of
quality legume inoculants. Initially Fitohoocmon was the proposed major private
company to be involved in the project for inoculant production as the profile supplied
by the company showed that it was producing soybean and groundnut inoculants.
However, the company soon withdrew from the project after proposing to produce
inoculants the same way as bio-fertilizers in Vietnam, i.e. based on unsterilised peat
and mixed with a huge amount of bio-fertilisers (up to 5 tonnes/ha). This does not fit
with the objectives of the project to produce and market high quality of inoculants,
applied at rates of around 1 kg/ha.

In the North, Private Business Ngoc Trung at Son La Town, Son La province (Son La)
and Viet A Nghia Dan Joint Stock Company at Nghia Dan, Nghe An (Nghe An) and
in the South the Komix company are interested in the objectives of the project and
they have become involved.

Inoculant production at Son La and Nghe An companies – SFI produces the broth
cultures and supplies them to the companies. The broth cultures are injected into
sterilized peat prepared by the companies. SFI transferred the technologies of peat
sterilization, injection, packaging and storage. The quality of inoculants has been
monitored by SFI throughout the production process. Final products were tested by an
independent quality control laboratory. Some pilot batch production was done and
inoculants were supplied for 90 ha of groundnut and soybean in Son La and Nghe An.

However, the cost of production was found to be high due to high cost of
transportation. Nghe An and Son La are around 300 km far from Hanoi where the
broths were produced by SFI. In addition, there is always the risk that the broths might
be damaged during transport from Hanoi to Son La.

The Komix company works in the project with OPI. There are 2 steps of the process.
In the first step they field-test inoculants produced by the institute, and market and
distribute them to farmers. Komix gets feedback of the potential of the inoculant based
on evaluation of biological and economic benefits of inoculation in the target