ts-6 beneficial indigenous microorganism products in microbiology
TRANSCRIPT
International Conference on the Role of Universities in Hands-On Education Rajamangala University of Technology Lanna, Chiang-Mai, Thailand 23-29 August 2009
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BENEFICIAL INDIGENOUS MICROORGANISM PRODUCTS
IN MICROBIOLOGY
Charunee Meechoui* Arissara Wanna Kingkan Wongjee
Kunnika Khuntong Sumalee �Promrukhachate and Sawit Meechoui
____________________________________ ABSTRACT
Indigenous microorganisms is a term of small beneficial microbes from that local area
(Tancho, 2004) which used in natural or organic agriculture system for soil remediation, composting,
water treatment and livestock etc. Beneficial indigenous microorganism products consisted of the
mixed cultures of beneficial and naturally living microorganisms that can be useful to increase
microbial diversity of soils and plants indirectly. �
To realize how to develop and produce the beneficial indigenous microorganism products
being suitable eastern wisdom technology, it’s necessary to integrate the knowledge of microbiology
biochemistry and fermentation technology.
This was studied the typical predominant microorganisms growth as microorganism ecology.
For the appropriated local wisdom fermentation was based on local submerged fermentation in fed-
batch process following as the local wisdom procedure. By this appropriated fermentation technology,
the living-fertile soil use natural inoculum and molasses as the carbon source. This methodology were
refer to Charunee and Sumalee (2550) that have been studied from batch culture fermentation of IMO
by viable count and direct microscopic count of motile spirillum, lactobacilli, and budding yeast.
The results showed a growth pattern of predominant indigenous microorganism according a time by
naturally submerge fed-batch process fermentation. The predominant microorganisms in the first 2 day
was nitrogen fixing living motile spirilla group that showed curve rod shape by gram stain which
shortly exponential phase stationary phase and decline phase. The second group of microorganism was
lactobacilli which grown 2-3 day after starting fermentation, and longer period growth phase than the
first, the lowest pH level was occurred in this period.The last predominant group was budding yeast
grown about 5-9 day the final fermentation which was film forming in appearance. The addition of soil
inocula and molasses every 10 day take a next cycle growth of microbial series that make microbial
population concentrated especially lactobacilli group which effective high quality of the indigenous
microorganisms inocula. In addition , t he ecological effects on the sequence of a microorganism
growth pattern were discussed.
Keywords : beneficial indigenous microorganisms products, �appropriated technology
Lampang Agricultural Research and Training Center, P.O. Box 89 Lampang 52000, Thailand Tel. 6654-342553 *Corresponding author. e-mail address : [email protected]
International Conference on the Role of Universities in Hands-On Education Rajamangala University of Technology Lanna, Chiang-Mai, Thailand 23-29 August 2009
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INTRODUCTION
The inoculum IMO preparation was done by using fertility soil in the forest, namely living
soil. It was fermented with molasses as carbon source. Fertile forest soil was as several microorganisms
groups. It may be done directly by using submerge culture fermentation method with molasses solution
for inoculums IMO culture, or by firstly adding different organic materials such as rice hull, rice bran
or cooked rice to ferment with the soil as followed solid state fermentation method, namely Baiyodo-
fermented soil. Then finally to do as submerge culture fermentation. During fermentation period, each
microorganism groups will growth as following inoculums soil microorganism ecology.
However, in Thailand, inoculums IMO culture product development and the appropriated
inoculums IMO technology , introduced from the east wisdom of Japan and Korea, should be modified
by integrating some knowledge bases, such as microbiology, fermentation technology and
biochemistry, etc.
This research was studied the natural IMO microbiological fermentation corresponded the
period of time by following batch and fed-batch process. In addition, the ecological results of the
sequent ordering microorganism growth were predicted.
MATERIALS AND METHODS
The local IMO inoculums preparation for this research was modified by both of Japanese local
wisdom method and the method from Tancho (2004). It was done as submerge culture by fed-batch
fermentation. Fertile forest soil, 200 g, obtained fulfilled microorganisms source was mixed with 5%
molasses solution, as carbon source for microorganisms growth in 30 liter plastic tank. Then the newer
mixed solution was added every 7 days until 1 month. And finally, the amount of each predominant
three microorganism groups; such as curve rod, lactobacilli and budding yeast were sampling everyday
to study their growth change under microscope under the gram strain counting method. It was
replicated 5 fields /slide. Their change was plotted to graph against the time.
RESULTS DISCUSSION AND CONCLUSTION
For fed-batch fermentation of inoculums IMO (in Figure 1.3), the different characteristic
growth of microorganisms were found. However, it was summarized only three predominant
microorganism groups were determined as 1.) curve rod bacteria, motile spirilla living former bacteria,
2.) lactobacilli group, and 3.) budding yeast group. This methodology were refer to Charunee and
Sumalee (2550) that have been studied from batch culture fermentation of IMO by viable count and
direct microscopic count of motile spirillum, lactobacilli, fission yeast and budding yeast. (From
those experiment, Charunee and Sumalee (2550) The figure 1.1 show typical time course fermentation
of indigenous microorganisms by direct microscopic count. And figure 1.2 show predominant major
groups of microorganisms isolated during typical time course fermentation of
indigenousmicroorganism products) From this experiment, firstly the response growth curve was
International Conference on the Role of Universities in Hands-On Education Rajamangala University of Technology Lanna, Chiang-Mai, Thailand 23-29 August 2009
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mostly the motile spirilla population increased and then decreased sharply in medium pH nutrient. This
bacteria growth changed quickly in exponential phase, stationary phase and decline phase of the growth
pattern. It was hypothesized that this bacteria growth firstly predominant and short growth period
because of nitrogen shortage condition at the beginning of fermentation. But it was able to fix nitrogen
in the air and then grew so fast and much more than the others.
0
20000000
40000000
60000000
80000000
100000000
120000000
140000000
160000000
0 1 2 3 4 5 6 7 8 9
Time (day)
cell/ml
motile spirilla
lactobacilli
Schizosaccharomyces
budding yeast
Figure 1.1 Typical time course fermentation of indigenous microorganisms by direct microscopic count.
International Conference on the Role of Universities in Hands-On Education Rajamangala University of Technology Lanna, Chiang-Mai, Thailand 23-29 August 2009
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Figure 1.2 Predominant major groups of microorganisms isolate during typical time course
fermentation of indigenous microorganism products
Left above : motile spirilla (1000x) 0.5x2.3-2.5 µm
Right above : lactobacilli (1000x) 0.4x10-20 µm
Left below : fission yeast , Schizosaccharomyces (1000x) 4x12 µm
Right below : budding yeast (400x) 4.5x12 µm
Bright field observation microscopy of motile spirilla living cell, the first major group
during typical time course fermentation. (left above) Gram’s stain microscopy of lactobacilli, the
second group of microorganisms which plays an important role in this system. (right below). The
fission yeast Schizosaccharomyces, unicellular eukaryote which is the third group in this ecosystem
grows together with lactobacilli. (left below) And the last group is budding yeast which grows after
another group is at final stage
Then this bacteria population had declined by death and composed to be organic nitrogen
International Conference on the Role of Universities in Hands-On Education Rajamangala University of Technology Lanna, Chiang-Mai, Thailand 23-29 August 2009
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source for the other microorganism living. In this research, it was isolated pure culture to classify. This
was spiral bacteria, movable and well growth in lack nitrogen source in media. For the secondary phase
of fermentation growth curve, it found lactobacilli bacteria population mainly increase in low pH media
condition. On the other, its growth made lower pH media. Finally, the budding yeast group, the third
generation microorganism growth was shown as the lactobacilli declining. Its growth as a thin film on
the surface of media could protect oxidation reaction by air to prevent acetic acid bacteria growth. It
may be the way to preserve the usage IMO for plant. On the other had, the observed predominantly
IMO growth pattern in the batch had change as a system and finally had low pH 3.9 for IMO
inoculums culture product. On the fed-batch fermentation, it affected to inoculums culture quality,
especially for lactic bacteria
REFFERENCES
Charunee Meechoui and Sumalee Promrukhachate. 2550.�Application Azospirillum and Rhodopseudomonas
of as Biofertilizers for Rice. Complete Research Report. Rajamangala University of Technology.
Tancho, A. 2004. Natural Agriculture. National Science and Technology Development Agency.
Bangkok. Thailand. 146 p. ISBN: 974-226-108-3 (in Thai)