The Ultimate Guide to Sterilizing Mushroom Substrate for Beginners (2024)

The cultivation of mushrooms at home is an enriching experience, offering fresh and delectable gourmet mushrooms that also offer numerous health benefits.

For novices, mushroom grow kits provide an exceptional starting point, and easy-to-cultivate varieties such as oyster mushrooms can be grown with minimal equipment. However, if you wish to cultivate more delicate or medicinal mushrooms that require nutrient-rich substrates, sterilizing your mushroom substrate is essential.

It is crucial to understand the difference between sterilization and pasteurization and how to sterilize your mushroom substrate using a pressure cooker. In the following sections, we will delve into these topics and provide step-by-step guidance on the sterilization process.

To prepare mushroom substrates, sterilization is a method that utilizes steam, pressure, time, and temperature to eradicate living organisms and spores.

Achieving a substrate sterilization requires a temperature of at least 250°F (121°C) for a minimum of 2 hours, which cannot be accomplished by boiling water or steam alone.

Water boils at 212°F (100°C) at sea level, and increasing the heat does not raise the temperature any higher. Thus, pressure needs to be added to elevate the temperature to 250°F (121°C) or above to eliminate bacteria and fungal spores.

Typically, a pressure of 15 pounds per square inch (PSI) is required to reach this temperature. Mushroom cultivators typically use a pressure cooker for sterilization on a small scale and switch to an autoclave, retort, or pressure canner as their mushroom business expands.

The requirement for substrate sterilization depends on the nutritional content and the type of mushrooms one aims to grow. Substrates with a high level of supplementation, such as manure, soy hulls, grains, and master's mix, as well as nutrient-rich substrates, are more prone to contamination.

These moist and nutrient-rich substrates attract not only mushroom mycelium but also other organisms like bacteria and mold, which can grow faster than the mushroom mycelium. Therefore, sterilizing these substrates becomes essential as it eradicates all living and dormant organisms and fungal spores, providing the best possible start for mushroom mycelium.

However, low-nutrient substrates like straw, sugarcane bagasse, coco coir, hardwood sawdust, and cardboard only require pasteurization. Our article on mushroom substrates offers additional information on different types of substrates and how to prepare them for cultivation.

The primary contrast between sterilization and pasteurization lies in their respective goals. While sterilization aims to eliminate all potential contaminants, pasteurization only eliminates most of the competing organisms.

There are various pasteurization methods that effectively reduce the number of competing organisms in low-nutrient substrates like straw and sugarcane bagasse, which provides fast-growing mushroom species an advantage.

However, for slow-growing mushroom species that require more nutritious substrates or longer colonization times, the best approach is to use sterilized substrates with minimal competition.

For those with some experience growing mushrooms who are interested in trying their hand at cultivating more difficult gourmet or medicinal species, sterilizing the substrate is a necessary step.

The most straightforward method for small-scale growers is using a pressure cooker that can maintain 15 PSI of pressure. To begin, gather the following items: a pressure cooker, jars or bags of prepared substrate, a wire rack or jar lids for the bottom of the pressure cooker, and water.

Mushroom grow bags are recommended since they are designed to withstand high sterilization temperatures and make growing mushrooms easier. However, jars may also be used for the substrate. Follow the steps below to safely sterilize the mushroom substrate:

Before using a pressure cooker for sterilizing mushroom substrate, it is essential to inspect it thoroughly to make sure all components are functioning properly. Look for any signs of damage such as dents, bulges, or cracks.

Some pressure cookers have rubber seals that may degrade over time, resulting in leaks. Check the seals carefully for any signs of damage.

Examine the cooker lid and ensure that nothing is blocking the vent pipe. Confirm that all screws are securely tightened and that the pressure gauge reads zero and is free of damage.

To prevent excess moisture from entering the substrate, you should fold mushroom grow bags to prevent steam from entering through the filter patch and cover any jar filters with foil.

Placing a metal rack or jar lids at the bottom of the pressure cooker and putting substrate jars or bags on top helps prevent melting or cracking from direct contact with the cooker's base.

Make sure to avoid having the bags or jars touch the sides of the pot, as it can cause damage to the containers. If necessary, line the lower sides of the pot with a cloth or kitchen towel.

To ensure that there is enough water for the entire sterilization process, you will need to add the appropriate amount of water to the pressure cooker.

Typically, it is recommended to add approximately 3 quarts (2.8 liters) of water, or enough water to cover the substrate bags or jars up to 1 to 2 inches (2.5 to 5 cm) from the sides.

As you gain more experience, you will be able to determine the exact amount of water required, but in the beginning, it is important to monitor and take notes.

Before you close the lid of the pressure cooker, make sure to weigh down any grow bags with a heavy plate or similar object. This is necessary to avoid the bags from obstructing the pressure relief valve on the lid of the pressure cooker.

To begin the sterilization process, put the lid on the pressure cooker and make sure it’s sealed tightly. Then, put the pressure cooker on the stove or gas cooker and turn the heat up to the maximum setting.

It may take some time for the water to boil and heat up the pressure cooker. When the pressure cooker gets hotter and the pressure builds up, steam will start to come out of the vent pipe.

Once the steam is coming out steadily and no longer spluttering, place a 15 PSI pressure regulator weight on the vent to regulate the pressure.

As heat continues to be applied, pressure within the pressure cooker will gradually increase until it reaches 15 PSI, causing the pressure regulator weight to move and let out steam.

Once this occurs, the heat should be adjusted until the steam has just enough pressure to cause the weight to move gently up and down.

Once you have achieved the right pressure, set a timer or alarm to keep track of your sterilization time.

After the sterilization process is complete, turn off the heat and leave the substrate to cool inside the pressure cooker for at least 8 hours.

Once the substrate is completely cool, it is ready to be inoculated with mushroom spawn. It's recommended to do this step in front of a laminar flow hood to minimize contamination.

Mushroom growers commonly use pressure cookers or autoclaves to sterilize their substrates since high temperatures above 250°F (121°C) cannot be reached without pressure.

However, it is possible to achieve similar results without a pressure cooker. To do this, growers must maintain high temperatures for longer periods, typically accomplished using a barrel steam sterilizer. These are ideal for small mushroom farmers who may not have the funds for an autoclave.

Since they operate without pressure, barrel steam sterilizers are less expensive, and it's possible to build one yourself. If you prefer to buy one, you can find ready-to-use barrel steam sterilizers online.

When using a steam sterilizer, the substrate is super pasteurized, which involves keeping it as close to 212°F (100°C) as possible for long periods. With a barrel steam sterilizer, the substrate must be kept between 194 and 212°F (90 and 100°C) for up to 18 hours, taking much longer than classic sterilization but producing nearly identical results and suitable for most mushroom species.

Sterilizing mushroom substrate in the oven is not possible, but it is possible to pasteurize it. This method is sometimes used for small amounts of substrate or casing material.

However, there are several challenges to consider when pasteurizing mushroom substrate in the oven. For example, you can’t use bags, so the substrate is often exposed to airborne contaminants when it cools down.

In addition, the oven’s dry heat may dry out and burn the substrate, reducing the nutrients available for the mycelium. If the substrate dries out too much, you’ll need to rehydrate it with distilled water to help prevent contamination.

Microwaving can be used to pasteurize small amounts of mushroom substrate, but it is not an effective method for sterilization since it cannot eliminate all fungal spores. Microwaves operate by exciting water molecules to produce heat and can only achieve temperatures up to the boiling point of water at 212°F (100°C).

Moreover, the substrate must have sufficient water content to be heated efficiently. Fungal spores contain low levels of water, making them challenging to heat, and steaming alone cannot produce high enough temperatures to kill them.

However, microwave steamers can be used to pasteurize small quantities of substrate and decrease the number of competing organisms.

The time needed to sterilize mushroom substrate depends on the quantity of substrate and the temperature used. In general, sterilization of substrate blocks in a pressure cooker at 250°F (121°C) takes between 1 to 4 hours.

The minimum time needed to sterilize small blocks is one hour, and for larger blocks, 3 to 4 hours is sufficient. If the blocks are sterilized for too long, they may become over-sterilized.

After sterilization, it's important to let the substrate cool before inoculation, which can take up to 8 hours. When using atmospheric sterilization in a barrel steam sterilizer, it can take up to 18 hours to sterilize the substrate, and the substrate must also cool before inoculation.


-Elimination of competing organisms


- increased yields through nutrient supplementation, and the ability to grow a wider variety of mushrooms on various nutrient-rich substrate


-high energy consumption


-higher cost of pressure equipment


-need for careful monitoring during the heating stage


-time-consuming process requires patience and dedication

Mushroom growers often use advanced equipment like pressure cookers, autoclaves, or barrel steam sterilizers to sterilize mushroom substrate. However, there's no need for expensive equipment or years of experience to start growing mushrooms at home. With simple methods and minimal equipment, you can grow gourmet mushroom species like shiitakes and oysters.

The Ultimate Guide to Sterilizing Mushroom Substrate for Beginners (2024)

FAQs

The Ultimate Guide to Sterilizing Mushroom Substrate for Beginners? ›

Foreign germs and competitor spores can be killed by hydrogen peroxide without harming mycelium. Soak the substrate for about an hour in water. Drain it, then thoroughly clean it in water before draining it again. Allow the straw substrate to soak in a hydrogen peroxide water bath for a day.

How do I sterilize my mushroom substrate? ›

Foreign germs and competitor spores can be killed by hydrogen peroxide without harming mycelium. Soak the substrate for about an hour in water. Drain it, then thoroughly clean it in water before draining it again. Allow the straw substrate to soak in a hydrogen peroxide water bath for a day.

What happens if you don't sterilize mushroom substrate? ›

Mushroom substrates are the perfect breeding ground for unwanted microbes. If not properly treated through pasteurization or sterilization, contamination will lead to wasted crops. Pasteurization simply reduces microscopic competition in a substrate while sterilization eliminates it entirely.

What pressure do you sterilize mushroom substrates? ›

Most substrates, grain, and agar need a Pressure of 15 PSI in order to completely kill off any potential contaminates. At this pressure, steam reaches a temperature of 121 deg C.

What is the spawn to substrate ratio for cubensis? ›

The recommended spawn to substrate ratio for growing mushrooms, such as cubensis, b plus mushrooms, or oyster mushrooms, is around 1:2 to 1:4. This means that for the psilocybe cubensis substrate recipe, one part of grain spawn should be used for every two to four parts of mushroom substrate.

Can you use hydrogen peroxide to sterilize mushroom substrate? ›

The method of ste- rilization with hydrogen peroxide H2O2 can be used as a successful method for sterilization of cultivation substrates for oyster mushrooms, taking into account the soaking for a longer period and increasing the period of time before ad- ding the mushroom spawn to the substrates to get rid of the ...

Does boiling water sterilize substrate? ›

Achieving a substrate sterilization requires a temperature of at least 250°F (121°C) for a minimum of 2 hours, which cannot be accomplished by boiling water or steam alone. Water boils at 212°F (100°C) at sea level, and increasing the heat does not raise the temperature any higher.

Is it better to pasteurize or sterilize substrate? ›

Pasteurization will not sterilize—or remove all contaminants—the way actual sterilization will. It really only weakens microorganisms, killing some pathogenic bacteria and reducing the level of substrate contaminants in the process.

How long does substrate stay sterile? ›

Sterile substrates do not require refrigeration, we recommend storing at room temperature. If kept at room temperature and out of direct sunlight, all sterile substrates can be stored for several months before use.

What temperature damages mycelium? ›

5), and thus can be attributed fully to the loss of physically adsorbed water. The increase in heat release between 100 and 200 °C can be attributed to the release of flammable low molecular weight volatiles. Mycelium starts to decompose at approximately 225 °C.

What temperature do you pasteurize mushroom substrate? ›

In general a compost substrate temperature of 140° F for 4 hours is adequate for a complete pasteurization. To insure a complete pasteurization it is suggested to have a minimum of 2 hour crossover time, where both the air and compost substrate at 140° F together.

How long does it take to autoclave mushroom substrate? ›

For example, hardwood sawdust in small volumes can be sterilised in under 20 minutes, while the same sawdust in blocks or large bags could take upwards of 2.5 hours. Jars of grain spawn can likewise take from 45 minutes to two hours.

What temperature should a Cubensis Monotub be? ›

Allow your bag to colonize in a cool location out of direct sunlight. The temperature for incubation shouldn't exceed 75 degrees F or dip below 55 degrees. After a couple weeks or so you should start to see mycelium growing on the grain. You'll notice the white mycelium growing outward from the injection point.

What is the best temperature for cubensis mycelium growth? ›

The optimal temperature for mycelium growth of both oyster mushroom species was obtained at 28℃.

How much worm castings to add to substrate? ›

Say if you're adding worm castings to a small container or seedling soil mix, you might only need a few handfuls. For seed starting, the suggested amount of worm castings to use is up to 50% of your soil mix. Whereas for houseplants, you'll need much less as they aren't fruiting plants.

How do you disinfect mycelium? ›

Steam cook-out should aim for 65-70˚C in the compost for a minimum of 8 hours. Although steam cook-out gives the best protection, infections can be transmitted from one crop to another at any time. Cleaning is needed before disinfection because disinfectants cannot kill mycelium fragments in compost.

How do you sterilize a Monotub? ›

Put some rubbing alcohol into a spray bottle and sterilize your entire tub. Spray your hands, your working surface, any scissors you might use to cut your spawn bag open etc. Spray the outside of your spawn bag and your bulk substrate bags and rub the alcohol around the entire bag.

How to sterilize mushroom casing? ›

The casing material can be disinfected by two methods: a thermal method (steaming) and a chemical method. At using the thermal method, it is recommended to treat the casing mixture with steam of low pressure at the temperature of 60-65°C for 6 hours.

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