COURSE: BED SCIENCE.

UNIT NAME: APPLIED MICROBIOLOGY.

UNIT CODE: SBT B406

LECTURER: DR MWAFAIDA

TASK: LABORATORY REPORT ON MUSHROOM PRODUCTION.

GROUP 3 MEMBERS.
NUMBER NAME ADMISSION NUMBER
1 SYLVIA NZARO MAPENZI E37/PU/0993/15

2 YUSUF OTENGO E37/PU/1059/15

3 GOWA MOHAMED E37S/PU/3652/15
4 LEONARD KIPYEGON KIRUI E37/PU/1038/15
5 ANTHONY JAMES WANJUI E37/PU/0991/15
6 VICTOR KETER E37/PU/1311/15
7 KIPLANGAT J RUTO E37/PU/0217/15
8 CHARLES MWENDA E37/PU/1020/15
9 ALBERT NYANGARESI E37/PU/2509/15
10 LEONARD SIWARENG E37/PU/1754/15
11 KENDUIYWO VICTOR KIPKIRUI E37/PU/1037/15

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 LABORATORY EXPERIMENT ON PREPARATION OF MUSHROOM SPAWN.
ABATRACT.
Mushroom cultivation has gained increased attention in recent years. Currently, only four types of
spawn, including sawdust spawn, grain spawn, liquid spawn, and stick spawn, are commonly available
for mushroom cultivation.
This limited spawn diversity has led to difficulty in selecting suitable inoculum materials in some
cultivation.
The experiment was carried using barley grain medium in which the mycelia from the mushroom was
grown in it.
The small shreds of mushroom was surface sterilized with 70% AIOH then it was placed on water agar to
facilitate growth of hyphae which was the transferred into second media PDA and incubated to produce
the plugs of mycelia which was required to be incubated on barley grain system to produce spawn which
can be sold for commercial purposes.

INTRODUCTION
Mushroom was belonging to the kingdom Fungi due to unique fungal characteristics (Song, 2004).
Mushrooms are being used as food and medicine since time immemorial.
According to Chang (1992) the protein value of dried mushroom has been found to be 30-40%
containing all the essential amino acids. Mushrooms constitute an ideal source to reduce body weight.
Mushrooms supply more protein per unit area than other crops (Gupta, 1986). Mushroom helps to
reduce serum cholesterol and high blood pressure (Mori et al., 1986). Ganoderma lucidum is renowned
for its medicinal properties (Willard, 1990) and often is associated with health and repercussion,
longevity, wisdom, and happiness (Stamets, 1990).
In Bangladesh, huge amount of agricultural wastes is produced annually, and are of no uses. These
wastes could be used as source of food i.e. substrate for mushroom cultivation.
Al Amin (2004) in his experiment revealed that the highest number of primordia and fruiting bodies of
Oyster mushroom was found in sterilized paddy straw.
However, a profitable mushroom cultivation on large scale also requires close attention,
experience and skill (Carluccio, 1989).

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 METHODOLOGY.
Apparatus and equipments.

 Water Agar and PDA

 Standard distilled water
 Forceps
 Isolation pins
 70% alcohol
 Clean bench
 Autoclave
 Microwave
 Sterile bloating paper
 Scarpel

Procedure for spawn culture

1.Preparation of mycelia.
 Mushroom was collected from the market
 Mushroom was stripped at the bottom into small shreds
 Mushroom was surface sterilized for 10 seconds with70% AIOH
 The surface sterilized mushroom was then rinsed through 3 sets of distilled water
 The mushroom was also Bloat dried on paper and was placed on water agar media
 The mushroom was then incubated for 2 days
 The grown hyphae were Pin isolated and transferred onto PDA media
 The hyphae were incubated for 3-5 days
 The plugs were then transferred onto barley medium.

Barley grain system

 20 g grains of barley were taken and soaked into 18 ml SDW

 Autoclaved and Allow to cool

 3-5 Plugs of 3-5 old day mycelia were added into barley grain system
 The plugs were then incubated on stationary conditions for 10 days.
 The set-up is not shaken and colonized medium is sold as spawn

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 OBSERVATION.
The small shreds obtained from mushroom button and placed on water agar media colonized the petri
plates by growing several single hyphae which can be seen when petri plates were placed against light.
Also, the cuts from single hyphae which was grown on PDA colonized petri plates with mycelia of 3-5
days in which we can observe their morphology.
The plugs of old mycelia which was placed in barley grain system colonized the sterilized bottle with
white and fluffy spawn which is indication of healthy colonized spawn.

DISCUSSION

There are 3 steps in spawn production;

 Raising pure culture

 Preparation of mother spawn
 Multiplication of spawn

Raising pure culture.

The shreds from the mushroom button were able to grow and colonized the water agar petri plates
since the media has necessary chemical substance which supports the growth of the hyphae.

The environment is sterilized thus it is free of contaminants such as bacteria and fungus which could
alter the growth of the hyphae, it also provides a conducive environment like in natural environment
where other mushroom grow.

The role of water agar was to produce hyphae which can be transferred onto PDA which is another
media for pure culture of hyphae obtained through process of pin isolation from water agar. When the
single hyphae are grown on this media allows the mycelia to grow after being incubated for 3-5 days.

Preparation of mother spawn

The barley grain was soaked in sterile distilled water and autoclaved to make environment to be
sterilized in that it kills the microbes that might cause contamination to the mycelia.

The old mycelia were placed at the middle of the barley grain to provide conducive environment for the
growth of colonies which are white and fluffy which are now the spawn which can be grown on the
favorable substrate of choice.

The purpose of grain spawn is to allow your chosen mycelium to multiply in a controlled environment,
before you let it loose on your fruiting (bulk) substrate.

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 Multiplication of spawn

Your grain spawn at this stage should be white and fluffy in the jar – this is a sign of healthy, colonised
spawn, here we can multiply our colonies by growing them in the substrate that allow the growth of
spawn forming multiple fruiting bodies which can be sold as premium commercial enterprise.

CONCLUSION

Mushroom cultivation has two main phases, spawn production and fruiting body production. The
mushroom seed is generally referred to as spawn. It is the propagating material used by the mushroom
growers for planting the beds.

Spawn is the vegetative mycelium grown on a convenient medium like wheat, pearl millet, sorghum, etc.
for raising mushroom crop.

It essentially involves preparation of pure culture of mushroom from tissues/ spores that is generally
maintained on any agar medium, followed by culturing on sterilized grains and further multiplied on
grains. The spawn thus comprises of mycelium of the mushroom and a supporting medium which
provides nutrition to the fungus during its growth.

REFERENCES.

1. Al Amin, M. A. 2004. Studies on mycelium, spawn and production of certain edible mushrooms.
M. Thesis, Department of Biotechnology, Bangladesh Agricultural University.
2. Carluccio, A. 1989. A Passion for Mushrooms. Pavillion Books Ltd, London.
3. Chang, S. T. 1992. Cultivation of Volvariella Mushrooms in Southeast Asia, In: Trop.
4. Mushrooms; Biological Nature and Cultivation Methods. The Chinese Univ. Press, Hong Kong.
pp. 135-156.

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