How to Produce Liquid Spawn
1 The liquid fermentation tank medium producing process.
1.1 add water to the tank inter-layer
First, add water into the liquid tank inter-layer, detail method :
connect the water pipe to the water inlet value (No.5) with the
silicone tube , meanwhile open the inter-layer outlet value (No.6) to
add water(avoid the high pressure in the inter-layer damaging the
inner tank), till water outflow from the drain valve.
1.2 the liquid medium formula
120L: corn flour : 0.75kg; bean flour: 0.5kg; defoamer: 20ml.
Mix the corn flour and bean flour with warm water, and no dough
ball, then add to the tank through the pump or funnel, it’s advise to
add the liquid to the tank sight glass, and add the defoamer 20ml,
then tighten the vaccination mouth.
1.3 liquid medium sterilization
Adjust the control box temperature to 125℃, open the
heating rods began to heat, before the temperature up to 100℃,
open the inter-layer water(No.6) valve to release extra pressure
and water in the inter-layer.
1.3.1liquid medium pneumatic stirring
When temperature is under 70℃, open the air pump to do the
pneumatic stirring to the liquid medium through the air tank and
filter shell to prevent liquid agglomerate. Steps to open the air pump:
open the filter shell inlet air valve(No.12)、the filter shell air outlet
valve(No.13)、the filter shell outlet valve(No.14), after open the air
pump(No.7), close the filter shell outlet valve(No.14), then open the
tank air inlet valve(No.15) and the vent air valve(No.2).
1.3.2 close the air pump
When the medium temperature in the tank is above 70℃,
close the air pump(No.7) as follows: close the air inlet valve(No.15),
open the filter shell air outlet valve(No.14), close the pump
power(No.7), then close the filter shell air outlet valve(No.13), the
filter shell air inlet valve(No.12), the filter shell air outlet
1.3.3 liquid medium sterilization
When the inter-layer water outlet valve(No.6) release hot
steam for 3 to 5 minutes, please close it. When the pressure meter
in the inter-layer(No.18) up to 0.05MPa, open the inter-layer vent
air valve(No.17), then open the filter shell air inlet valve(No.16) in
the inter-layer, open the tank air inlet valve(No.15), open the tank
air outlet valve(No.2), when you feel the inlet air tube hot, close the
tank air outlet valve(No.2). When the tank pressure meter(No.3) up
to 0.15MPa, start timing, open the tank air outlet valve 1/3(No.2),
and adjust the temperature according to the pressure for 30 to 40
minutes to sterilize.
After timing, adjust and control the temperature to 25℃, then
close the heating rod, close the tank air inlet valve(No.15), close
the inter-layer air inlet valve(No.16), close the filter shell inter-layer
air outlet valve(No.17), open the filter shell air outlet valve(No.13)
and filter shell air inlet valve(No.12).
1.3.5 drain hot water in inter-layer
Open the tank air outlet valve(No.2), drain the hot water in the
inter-layer through the inter-layer water inlet valve(No.5) (tighten
the silicone tube, fix the hot water outlet well and avoid burning
people ) until the pressure data in interlayer pressure meter( No.18)
down to 0.
1.4liquid medium cooling
Open the inter-layer water outlet valve(No.6), connect the
inter-layer water inlet valve(No.5) to the water tube through silicon
tube for cooling. When the tank pressure meter(No.3) below
0.05MPa, open the air pump(No.7) (prevent the tank causing
pollution in the cooling process, and also make it lower water cool
faster). Steps to open the air pump: open the filter shell air outlet
valve(No.14), open the filter shell air outlet valve(No.13), open the
air pump(No.7), close the filter shell air outlet valve(No.14), open
the tank air inlet valve(No.15), then adjust the tank pressure range
from 0MPa to 0.05MPa through the tank air vent valve, when
temperature cools down to 25℃, then begin inoculation(the
temperature varies according to different species ).
2.1liquid strains culture medium
Sawdust(need screen):500g, bran:100g, gypsum:10g. Mix all
these raw material with water to the 500ml triangle bottle, the
mother culture method is same as the solid seed production
2.2 production of sterile water
Make 700-800ml sterile water by portable pressure cooker,
keep the pressure at 0.15MPa for one hour, after the sterile water
cool down, inoculate the strain.
2.3 inoculate the strain to the sterile water
2.3.1 inoculation appliance
Alcohol lamp(one), 75% alcohol bottle(about 30ml),
vaccination hook, screwdriver.
Rotate the triangle bottle with liquid strains culture medium on
the alcohol lamp flame evenly, and place the inoculation appliance,
sterile water, and a bottle of penicillin(400million units) in the air
purifying workbench for 30-40 minutes to sterilize.
2.3.3 vaccination preparation
Wipe hands in 75% alcohol sponge, and use the alcohol lamp
flame to burn the sterilization vaccination hook(after sterilization,
can put in 75% alcohol to cool down), use the screwdriver to
remove the penicillin aluminum cover(do no cock the inside skin )
Under the protection of alcohol flame, use the sterilization
inoculation hook to remove the surface of triangle bottle, stir the
middle and low part of strain, and pour to the sterile water 2-3times
in alcohol lamp flame. Then pour the penicillin.
2.4strains to tanks
2.4.1 produce the fire circle
Wind gauze to the iron ring (the inside diameter is slightly
bigger than the inoculation diameter), then pour 95% alcohol.
Open the tank air outlet valve(No.2) to make the pressure
under 0, light the flame ring then close the tank air vent valve(No.2),
open the inoculation mouth(No.1) quickly, steadily,and softly to
vaccinate the strain(when open the inoculation mouth, the tank
pressure may rise again, you can vent the air through the tank air
vent valve(No.2)), tighten the inoculation mouth(No.1). Open the
tank air outlet valve(No.2), the pressure will be 0-0.05MPa, then
enter the training stage.
3.liquid strain testing
Due to edible fungus varies, so the training time is also
different, generally, after four days of mushroom strain inoculation,
it needs strain testing, firstly, use the alcohol lamp flame to burn the
inoculation valve(No.4) for 30-40 seconds, and ignore the initial
small flow of liquid strains, then inoculate directly on to the vitro
medium slant, pour the water out along the slant, and leave small
amount of fungus ball for training(also can cultivate the material by
the petri dish). After inoculation, dry up the inoculation valve(No.4)
with alcohol lamp flame to avoid bacterial contamination.
Notes： the filter(No.11) need pack into the filter shell(No.10),
install properly to ensure no leakage. Assistant products: add small
amount of meal cleaning solvent(for lubricate) filter above the
sealing ring, and then tighten and press.