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Simplified and Lower Cost Methods for Culinary-Medicinal Mushrooms

Cultivation

Article  in  International Journal of Medicinal Mushrooms · January 2012

DOI: 10.1615/IntJMedMushr.v14.i3.80 · Source: PubMed

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 Diversified Agriculture Part 1: Simplified and Lower Cost Methods for Mushroom
Cultivation in Africa

Britt M. Gianotti1, Matthew P. Cleaver1, Phillip D. Cleaver1, Cody Bailey1and John C. Holliday1

1. Aloha Medicinals Inc 2300 Arrowhead Dr, Carson City, NV 89706

Correspondent Author: britt@alohamedicinals.com

ABSTRACT

Mushroom cultivation in more developed countries has evolved from an art into huge agri-
business by way of the most innovative production technology and biotechnology available.
However, there are still less-developed areas of the world where access to these technological
advances are either unavailable or too expensive to utilize.

West Africa exemplifies this problem, and a low cost mushroom production program would have
compounded benefits for the region. This process would allow the use of secondary crops to be
produced in a short time frame from agricultural by-products derived from the primary
agriculture of the region, such as millet and sorghum straw, cassava peelings, or virtually any
other cellulose source. These and other potential raw materials are currently being disposed of as
waste. After the production of edible or medicinal mushroom crops are harvested from these
agricultural wastes, the residual mushroom substrate represents a bioconversion from the non-
nutritional cellulosic material to an edible fungal matrix of high protein content which can be
utilized as nutritious cattle and goat fodder as well as feedstock for tilapia farming. This
diversified approach results in three successive cash crops from any agricultural operation where
previously there had been only the one primary crop.

The oyster mushroom complex (Pleurotus ostreatus and other Pleurotus and Hypsizygus
species) appear to be the best candidates for production in the West African climate. These
mushrooms are primary cellulose decomposers, and can grow on almost any plant material
substrate including banana waste, coffee residue, sugar cane baggasse, paper or cardboard waste,
river grass, sawdust and nearly all other agricultural waste.

The purpose of this paper is to present low-cost, low-technology methods applicable to small-
scale village mushroom production. The process obtained from this research represents the least
training and smallest capital and infrastructure investment to implement a mushroom farming
operation, while suggesting the easiest system to grow locally acclimatized mushroom strains for
a large and valued mushroom crop in the shortest possible time with minimal financial risk for
the farmers.

INTRODUCTION:

13
A universal need for all mankind is food. Reliable supplies of low cost, nutritious and readily
available food for the population are the key items to a stable and healthy society. Until this basic
need is met, poverty alleviation and economic development cannot take place. To achieve the
goal of stable, healthy societies, mankind over most of the world has evolved from hunter-
gatherer societies to agriculture, aquaculture and animal husbandry for production of their
necessary foodstuffs. These farming endeavors have developed over centuries of social
evolution, originally utilizing local plants, fish and animals as the raw inputs to make these
production systems work. However, in the last century or two this has begun to change, with
plants and animals from far and wide being imported into new locals as superior candidates for
cultivation over the plants and animals found originally in that region.

This paper presents the first step in this expanded cultivation scheme, which is Mushroom
Production through utilization of low cost, technologically simple methods that can be easily
implemented at the village level in any country, be it highly developed or the poorest country on
earth. Our main goals in this research were to develop real world solutions to the problems faced
in implementing these cultivation systems. It is easy to imagine a system where if you have
capital resources of X, Y and Z, you can achieve great results, but what about those peoples that
do not have even the minimum resources of X, Y and Z? It is to these people that we have
directed this research. Using only agricultural waste which we have diverted from the disposal
stream, and commonly available tools, equipment and supplies, we will show in this paper how
anyone of even the most modest means can produce an economically viable crop of wholesome
and nutritious food with which to feed their families and villages; the surplus of which can be
exported to others as a cash crop. At the end of the first step of this diversified agriculture
program the remaining mushroom waste and spent substrate can be utilized as nutritious fodder
for goats, cattle, horses, camels, sheep, and fish farming. This allows a third crop, where initially
there was only one. To put this in perspective, consider the typical case of a cassava farmer in
Africa: the cassava is grown and harvested, and the plant waste and cassava peeling are dumped
back on the ground. The next crop to be harvested by the farmer is the next crop of cassava. By
implementing the system proposed here, the cassava peelings and plant waste becomes the
feedstock for the mushroom cultivation which yields a second crop (mushrooms) in
approximately three weeks, long before the next crop of cassava would be ready for harvest. The
waste materials from the mushroom production then becomes a highly nutritious feed for
animals and fish such as goats, cows and Tilapia, producing a third crop, where there originally
had been but the one primary crop. This increase in efficiency with little additional input of labor
or capital has perhaps the greatest potential for elevating the economic status of the rural farmer
across much of the world.

Mushroom cultivation represents a very basic natural process, that of fungal decay. In nature the
primary function of plants is to act as the molecular assemblers. Plants take simple compounds
like water and carbon dioxide and using sunlight as energy they assemble these into complex
organic compounds such as proteins, carbohydrates and fats. Upon the death of these plants, the
fungi move in and become the molecular disassemblers. That is to say the fungi take these
complex compounds like the proteins and carbohydrates and disassemble them back into the
simple compounds which we started with, those compounds like water and carbon dioxide. As
these simple compounds are released back into the environment by the fungi, they become

14
available as raw materials for the next generation of plant growth. In the process of running
through their life cycle, the fungi reproduce as do all other living things. In the case of fungi, the
reproductive organ can take several forms. One particular class of fungus, the basidiomycetes,
produce a large fleshy reproductive organ (or fruitbody as it is named) which we know more
commonly as the mushroom. Many of these mushrooms are choice edibles, and in fact represent
very high quality nutrition, often equaling or exceeding meat in quantities of protein and
essential amino acids. [1]

The basis of mushroom cultivation is the breakdown of cellulose. The cell wall structure of
virtually all plants is a fibrous structure composed of cellulose and hemicellulose, surrounded by
a structural compound called lignin. The lignin wraps around the cellulose fibers like plastic
wrap. This makes for a very strong structure, allowing a tree to stand upright for hundreds of
years. The cellulose and hemicellulose are made of sugars which are great sources of food, but
these are protected by the lignin wrapping, which is a very stable compound and difficult to
breakdown. Only a few organisms can breakdown the lignin and utilize it as a food source, thus
exposing the underlying cellulose and hemicellulose for food use by other organisms. The best
known and most effective of these lignin-breakdown organisms are known as the white rot fungi,
of which the Oyster mushrooms are the prime examples. Oyster mushrooms are a closely related
species complex, comprised of many species within the genera Hypsyzygus and Pleurotus. Many
different species of Oyster mushrooms have evolved in different locations around the world, and
have become more or less specialized in degrading different raw material cellulose sources (the
substrate) at different temperatures, oxygen and light levels (acclimatization).

A word on species and strains: There are many “Oystereque” mushrooms, with similar growth
characteristics, morphology (looks) edibility, flavor, shelf life, etc. Some of these are very
closely related to each other, even being given the same genus and species name such as the
common tree oyster, Pleurotus ostreatus. However, not all Tree oysters are completely alike
even though they may have the same genus and species name. For example, one may be growing
on an oak tree in the temperate region, while another is growing on an oil palm in the tropics.
These two mushrooms may be the same species but they are different strains. They will each
have evolved different enzymes to degrade their respective substrates, and they will have
different growth parameters having to do with the temperature, humidity and sunlight intensity in
the regions where they evolved. To try to grow the northern climate oak mushroom on a palm
tree in the tropics would yield very disappointing results. Then there are other Oysteresque
mushrooms that are not quite as closely related, but which still share most of the characteristics
with the other Oyster mushrooms. These have names like Hypsyzygus ulmarius, the Elm Oyster,
so named because it usually grows on Elm trees. This does not mean that it will ONLY grow on
Elm trees, just that this is the usual native habitat.

Choosing the best species and strains of Oyster mushrooms is the single most important step in
creating a successful mushroom cultivation program. There is no one strain that is any better than
another strain, it all just depends on the climate and the substrate and the cultivation methods
used. The only way to determine the best species and strain for growing in any particular
situation is to experiment with several types of Oyster mushrooms and record the production
time and yield for each strain in that particular circumstance. Generally, a local strain of

15
mushroom isolated from nearby the growing location is the best choice, as this local strain will
have become acclimatized to the local environment and the local competitors for the food source.

For the purpose of this research, we choose several strains of oyster mushrooms to indicate the
possibility of different strains being utilizable in this cultivation method, and to show that the
results obtained in this research were not strain specific. It should not be assumed that the results
shown here will necessarily transfer 100% to another location, strain or substrate, although the
general guidelines shown here will certainly apply. Anyone desiring to follow this methodology
of mushroom cultivation is encouraged to experiment with different species and strains, and to
include locally acclimatized strains whenever possible. In some cases such as northern climates,
it is not unusual to see several different species or strains cycled through a farm over the course
of the year as the seasons change. This way, one can assure the maximum yield both in the hot
season and in the cold season by selecting appropriate strains according to prevailing
temperature. Likewise, it is often preferable to cycle different strains through the farm seasonally
depending on substrate availability. If you have a strain that grows well on straw, you should use
that one at grain harvest time when straw is readily available, but if your only substrate is river
grass or cassava peeling, by all means be flexible in switching to strains particular to these
substrates so as to maximize your harvest.

The most basic concept of mushroom cultivation is that we need to produce an environment in
the substrate that is selectively preferential to the growth of our target species of mushroom, and
less amenable to other types of microorganisms and pests. This involves sterilization (completely
killing any other organisms that are present in the substrate which would compete with the
mushrooms for utilization of the substrate as food) or pasteurization (killing off the majority of
competitive organisms). Mushroom cultivation in more developed countries has evolved from an
art into huge agri-business by way of the most innovative production technology and
biotechnology available. This usually means the use of large pressure chambers and high
temperature steam for sterilization of the substrate. However, steam and the associated
equipment are both expensive and technically demanding in terms of training and education.
There are many less-developed areas of the world where access to these technological advances
are either unavailable or too expensive to utilize. In those areas it would be much more practical
to look at ways to pasteurize rather than sterilize the substrate, and by methods other than the use
of steam. This can be done in many ways, such as using commonly available substances such as
soap or hydrated lime to pasteurize the substrate. The mechanism by which these chemicals work
is through the rapid change in osmotic pressure, causing the majority of microorganisms present
to burst from the change in osmotic pressure. Once the substrate is treated in this way, and the
majority of the microorganisms are killed, the substrate is suitable for the introduction of our
target species, the Oyster mushroom.

Another potential treatment method is simply to soak the substrate in water for a week or so.
This results in a rapid bloom of the bacteria that rapidly consume the readily available simple
sugars. Once the sugars are exhausted after about 5 days of fermentation, the bacterial bloom
dies off due to lack of any more simple nutrient availability. This leaves a substrate which is
more selective to the higher cellulose degrading organisms such as Oyster Mushrooms. This is
the simplest method of substrate preparation; however it is the least effective of any methods we

16
have tried. It does work, but the chemical treatment methods are much more effective and not
considerably more difficult or expensive.

MATERIALS AND METHODS:

Substrates used:

Unshredded wheat straw (whole stalk), shredded wheat straw (reduced in a hammer mill to small
particle size). Wheat straw is the post-harvest stalk of Triticum aestivum and Ground maize cob
(Zea mays) were used as the substrates. All straw was obtained from S&W feeds, Carson City,
NV. Ground maize cob obtained from Benson’s Feed and Tack, Carson City, NV

Species and Strains used:

Although there are several fungi capable of white rot degradation, the following genus and
species in the Tricholomataceae family were used in this experiment: Pleurotus pulmonarius AX
strain, Hypsizygous ulmarium ELM 1 strain, Pleurotus D’jamor PDJ strain, Pleurotus Sajar-caju
PSAJ strain, and Pleurotus ostreatus TL strain. All cultures used in this research are
commercially used in mushroom farming, and were all provided by Aloha Medicinals Inc. of
Carson City, Nevada.

Spawn used:

Spawn is the seed stock which is used for inoculating the substrate for mushroom production.
The spawn for this research was generated under the usual conditions which will be known to
anyone familiar with spawn production. Spawn is generally grown on grain or sawdust. The
spawn substrate used for this project was White Sorghum grain, which was cooked with a
measured quantity of water to achieve a moisture content of approx. 55%. A small amount of
ground oyster shell was added to control the pH. Approx. 400 g oyster shell per 100 kgs of dry
grain was added, along with 102 kgs of water, which resulted in a pH of 7.0. After cooking with
water and oyster shell, gypsum was added to keep the individual grains of sorghum separated
and reduce the possibility of anaerobic conditions forming in the spawn. The cooked, gypsum
treated grain was filled into glass jars of 1 liter size until jars were approx. ¾ full, which
measured an average of 454 g of weight per bottle. The top of the jars were covered with a filter
made of thick paper and a metal lid screwed over the filter paper. The metal lid has a single
central hole drilled, measuring approx. 25 mm in diameter. This allows gas exchange of the
spawn substrate while eliminating the introduction of any foreign organisms during the spawn
grow-out period. These filled, capped jars were then sterilized in an autoclave at 17 psi steam
pressure (approx. 1.2 bar) for a period of 2.5 hours. After the sterilized jars were cooled
overnight, a small piece of inoculum of the appropriate strain was introduced into each jar under
sterile conditions, as will be known by anyone familiar with the art of spawn making. The
inoculated grain jars were then grown for 10 to 12 days before use, which resulted in completely
colonized grain of only the target species of mushroom. The short growth period of 10 to 12 days
means there was some percentage of unconverted grain starch remaining in the spawn, which
becomes a nutritional amendment to the final mushroom substrate, raising the nutrient level of
the straw without raising the risk of contamination. The concept of using young grain spawn as

17
both inoculum and nutrition amendment is important to the simplified cultivation process
presented here. Simply adding a rich nutrient amendment to an otherwise poor nutrient base like
straw or maize cobs will increase the risk of contamination by unwanted competitive organisms,
which would lead to reduced yield of the target mushroom fruitbodies. By using the grain spawn
as mentioned here, the grain is already fully colonized by the target mushroom strain, which in
turns eliminates all competitors within that jar, while still providing a rich nitrogen and starch
base for quick initial growth in the final fruiting substrate for the mushroom organism.

Spawning:

Spawning is the term used in inoculating the final substrate in order to grow mushrooms. In this
research 2.73 kg (6 pounds) of treated straw was added to a growing bag for each strain and
treatment type. All treatment methods and strains were done in triplicate time three, or in other
words a minimum of nine bags were used for each treatment method and strain, insuring
statistical reliability of the data generated.

The growing bags in this research were spawned with the addition of 454 g (one pound) of
spawn per bag, added alternately with a layer of straw as the bags were filled. The layers of straw
were approx 50 mm thick, then a sprinkling of spawn, before adding the next layer of straw,
followed by another layer of spawn until the bags were full. This represents a spawn to substrate
ratio of approximately 1:6. This is considered a heavy rate of spawning, but any spawn ratio
from 1: 5 up to 1: 40 can be used. It is generally considered that the higher the rate of spawning
(the more spawn used per bag) the better the results in consistency and yield, while a lower rate
of spawning (less spawn per bag) is cheaper, but with a correspondingly lower yield of
mushrooms per bag. The spawn rate chosen in farm practice is usually a reflection on the price
and availability of spawn in the local region. Whenever possible, it is always better to use more
spawn rather than less.

Growing Bags used:

Clear polyethylene bags of approx 300 mm x 700 mm x 2 mil thickness were used as growing
containers for this experiment. The bags were filled with substrate and spawn, with the excess air
squeezed out and the bags sealed by twisting a short metal wire or twine around the top. After 48
hours post-filling, the bags were ventilated by puncturing repeatedly with a knife blade. The
knife blade was 20 mm wide and ventilation holes were made on a random spacing approx 100
mm apart all over the surface of the bag. These holes both allowed gas exchange and a place for
the mushrooms to erupt from when fruiting.

Substrate Treatments:

A control batch of substrate for comparisons was made by first soaking the whole straw for 24
hours in barrels of water, and then draining the straw by setting it on racks and covering by
newspaper for two hours resulting in approx. 60% moisture content in the straw. The straw was
then treated with steam at atmospheric pressure for 2 hours (maintaining ~100 degrees C). The

18
bins were then tipped so that the excess water condensed from the steam treatment could drain
off for two hours while cooling. This straw was filled into growing bags and spawned in an
identical fashion to the experimental bags and grown in the same room and under identical
conditions throughout the trial period. This method of steam treatment is well known in the
mushroom industry and is well understood by anyone familiar with mushroom farming, and so
forms the comparison for all other substrate treatment methods. Straw treated with steam had no
detergent, lime or other substances added.

A second method of substrate conditioning used was the natural fermentation of one bale of
wheat straw (approx. 30 kgs) in water. The straw was soaked in barrels containing 200L of clean
tap water for seven days with no other substances added. The straw was removed from the
barrels after seven days and placed on racks for two hours, covered with newspaper in order to
drain excess water and allow out-gassing and aeration of the straw. As can be imagined, after
soaking this straw for seven days in the sun it was a stinky, sticky mess, but actually had a fairly
low microorganism load due to exhaustion of the simple sugars and other readily utilizable food
sources upon which the first generation of decay organisms depend. This fermented straw was
layered with spawn (454 g bottle) in four plastic bags for each test strain until a total weight for
each bag was 2.73 kg (6 pounds). When the bags reached the proper weight, they were
compressed by hand and tied off using segments of light metal wire. These bags were kept
outside in the shade for two weeks of observation and then moved inside a climate controlled
grow room. While this treatment method did produce edible mushrooms, it did not result in
commercially viable quantities of mushrooms being produced. This method of substrate
preparation is not very effective when compared to the other methods utilized in this trial, and
the data on this treatment method is not shown in the results.

Two more bales (approx 30 kgs ea) of unshredded wheat straw were soaked in several barrels
containing 200L water each and treated with one of the following substances for 24 hours:

1. 355 g per 200 L water of hydrated lime (Calcium hydroxide )(Rockwell Lime Co.
Manitowoc, WI, 54220)

2. 710 mL per 200 L water 5% Chlorine Bleach (Clorox Co. Oakland, CA 94612

4. 240 g per 200 L water Tri Sodium Phosphate (Savogran Co. Norwood MA 02062)

3. 120 g clothes washing powder per 200 L water (Tide brand clothes washing detergent, Procter
& Gamble, Cincinnati, OH)

After soaking in the above solutions for 24 hrs, the straw was laid out on top of large metal racks
and drained in the same manner as the fermented straw.

Shredding Straw and Maize Cobs:

When shredding the straw for the second portion of this experiment, dry straw was run through a
50 HP Jacobsen Hammer Mill for shredding. The shredded straw was soaked and processed in

19
the same fashion as the regular unshredded straw. The bags used for the shredded straw were
hand compressed more tightly than the regular straw after inoculating each, due to the smaller
particle size of the substrate. This additional compression was possible due to the increased
density of the shredded straw.

In addition, a drum containing 118 mL bleach was mixed with 200L water and filled with
shredded maize cob. Once the maize cob was sufficiently drained of excess water by straining
through small mesh chicken wire, the material was filled into identical growing bags and
spawned. For spawning, bottles containing 454 g of spawn were mixed intermittently with the
maize cob in a manner similar to that described for the straw. The maize cob substrate treated
with bleach resulted in yields very similar to the straw substrate. The outcome for the maize cob
substrate is not shown in the results simply in the interest of conserving space. It is notable that
the information regarding straw can be used representatively with maize waste and with most
other cellulosic agricultural waste.
.
All of the shredded straw bags and the maize cob bags were compressed by hand and tied with
string, wire or rubber bands. They were then placed on racks in the same growing room as the
unshredded straw bags. The growing room was maintained at a temperature of 21 to 25 degrees
C. Humidity was maintained with the use of humidifiers from 80% to 90% RH. Light cycle was
natural shaded light, approx 12 hrs per day of light and dark.

Picking Procedure:

Mushrooms were checked daily and picked at specific times in their growth cycle: eg. When the
edges of the caps were flattening out or showing signs of slight curling. The mushrooms were
picked, weighed, and put into a refrigerator. Pictures and weights were taken and recorded at
each harvest.

Observation sheets:

Observations were taken bi-weekly in order to note the difference and similarities between
growth for each treatment method and individual strains. Characteristics noted included the
amount of straw sprouts, bacterial contamination, competitive mold contamination, mycelial
growth, number of primordial formation (not yet matured fruitbodies called pins) and fruitbody
clusters, and the weight of the harvest.

Bag disposal:

Bags that became overly contaminated by bacteria or competitive molds, were weighed using an
accurate lab scale (Yamato Corp. Colorado Springs, CO 80906) and disposed of immediately to
avoid contamination of the remaining bags. After the remaining bags passed two months of
active harvest, they were also weighed and disposed of.

Biological Efficiency equations:

20
Biological efficiencies (BE) for each strain were attained by dividing the average fresh weight
totals for that treatment method and strain type by the average dry weight of substrate and
multiplied by 100. This resulting figure is given as a percentage, with the figure of 100% BE
equal to 2.5 kgs of dry weight substrate yielding 2.5 kgs fresh weight mushrooms. In this study,
it was not uncommon to see BE’s in the neighborhood of 150% to 180%.

RESULTS AND DISCUSSION:

In the third world, protein-energy malnutrition (PEM) affects 500 million people and kills 10
million annually. [2] To avoid ill health from certain diseases, and death caused from nutritional
deficiency, daily dietary protein consumption is essential.[3, 4]

In countries that suffer from widespread protein deficiency, available potential food sources are
generally full of plant fibers from the terrestrial biomass. Wood and straw make up about 80% of
this biomass, which equates to approximately 800 billion tons worldwide, an extremely large
reservoir. [4, 5, 6] Although four stomached mammals (ruminants) such as cows, goats and sheep
produce the enzymes necessary to digest the cellulose present in plants such as grasses and hay,
they are unable to directly utilize the biggest cellulose reservoir; woody biomass, which remains
untouchable because of the lignin that is structurally wrapped around the cellulose. Lignin is a
complex chemical compound found in plant cell walls that is cross-linked with other cell wall
components, and acts as an integral part of the secondary cell walls of plants[7] and some algae[3].
As a biopolymer lignin is unusual because of its assorted composition and lack of defined
primary structure. Lignin is most commonly noted for support through strengthening of wood
(xylem cells) in trees [5, 6, 8] and is generally only degradable by highly specialized enzymes
associated with very few organisms.[8] Therefore, this molecule is also generally associated with
reduced digestibility which helps defend against pathogens and pests.[9] In order to make use of
this cellulose, the lignin first needs to be stripped, leaving the cellulose exposed. As mentioned
above, all forms of White-rot fungi such as Pleurotus ostreatus are able to oxidize the non-
digestible lignin, setting free the cellulose and giving both the fungi and other potential
consumers such as ruminants ready access to this significant nutrient reservoir.

As will be seen from the graphs and illustrations, this experiment clearly shows the practicality
of raising mushrooms on low cost (or free) agricultural waste such as straw, that has been treated
by simple and low cost methods such as soaking with a bit of lime or washing powder. In fact,
the yields seen with these low cost treatment methods were not substantially different than the
results seen in the steam treated control group. The biggest factor in yields was the density of the
substrate, with shredded straw showing considerably higher yields than was seen with
unshredded straw.

The results of this experiment show no significant difference between the mushroom
marketability and characteristics such as fruit body, yield, taste, color, smell, etc. of the low cost,
low technological methods described here as compared to the high technological steam-based
methods used with most mushroom farming in developed nations today.

Implementing a low tech mushroom farming approach and low cost mushroom production
program in West Africa would have compounded benefits for the region. This process would

21
allow the use of secondary crops to be produced in a short time frame from agricultural by-
products derived from the primary agriculture of the region. These and other potential raw
materials are currently being disposed of as waste. After the production of edible or medicinal
mushroom crops from these agricultural wastes, the residual mushroom substrate represents a
bioconversion from the non-nutritional cellulosic material to an edible fungal matrix of high
protein content which can be utilized as nutritious cattle and goat fodder as well as feedstock for
tilapia farming. This diversified approach results in three successive cash crops from any
agricultural operation where previously there had been only the one primary crop.

The oyster mushroom complexes used for this experiment appear to be one of the best candidates
for production in the West African climate. These mushrooms are primary lignin/cellulose
decomposers, and can grow on almost any plant material available for substrate including banana
waste, coffee residue, sugar cane baggasse, cassava peelings, paper or cardboard waste, river
grass, sawdust and nearly any other agricultural waste.

Other studies have been successful in revealing semi-low-tech methods for recycling agricultural
waste and utilizing white-rot fungi for the production of fodder for ruminants which showed fast
and reliable conditioning of the substrate. [10] This type of study was also able to help in the
revitalization of the local grazing areas from being arid and dry from overgrazing to lush and
green. To take this idea one step further, we explored different detergent solutions as a method
for pasteurization to find which will provide the best protection against outside contamination,
while still producing the highest yield. In this way farmers will be able to produce less expensive
fodder for their herds while having a commodity that will feed and support their families as well
as raise their economic status and maintain the land. This type of technique has been shown to
decrease the land needed to maintain healthy livestock and increase employment availability. An
addition to these obviously stated facts, the first harvest of oyster mushrooms occurs only 18 to
21 days after inoculation. There are few other crops that can give the farmer an income in less
than 3 weeks.

Many of the graphs employed are calculated using the values found for biological efficiency
(BE). BE is the amount of growth determined by a comparison between the dry weight of
substrate compared to the fresh weight of the mushrooms harvested. BE indicates the total
percentage of growth in terms of mushroom fruitbodies. The figures below indicate a significant
increase in BE when using shredded straw as the substrate for all the treatment types. In fact,
substrate density appears to be the greatest determining factor in mushroom yield, at least for
straw substrate and the strains of oyster mushrooms tested in this experiment.

22
Figure 1a shows the steam treatment comparison between unshredded and shredded straw. Steam
treatment was used as a control in this experiment for both shredded and unshredded straw as it
is a known and regularly used method for substrate sterilization. The AX strain was found to
have an efficiency of 129.8% on shredded straw and 34% on unshredded straw, which is a 95.8%
difference. The ELM 1 strain had an efficiency of 175.6% on shredded straw and 43.3% on
unshredded straw, a 132.3% difference. The PDJ strain had an efficiency of 102.3% on shredded
straw and 24.6% on unshredded straw, a 77.7% difference. The PSAJ strain had an efficiency of
119.1% on shredded straw and 30.5% on unshredded straw, an 88.6% difference. As seen in this
graph, the biological efficiency for any of the Pleurotus strains using the shredded straw as a
substrate and steam as the treatment gives a substantially higher yield then using unshredded
straw as the substrate.

Figure 1a: Steam growth comparison

Steam Growth Comparison

200.00%

180.00%

160.00%

140.00%

120.00%

BE Shredded Straw
100.00%
Regular Straw

80.00%

60.00%

40.00%

20.00%

0.00%
AX ELM PDJ PSAJ
Strain

Figure 1a shows a direct comparison between the Bio efficiencies of shredded straw to regular unshredded straw for each of the representative
Pleurotus strains, AX, ELM, PDJ, and PSAJ using an autoclave for the steam sterilization method (122 °C 1 bar pressure).

23
Figure 1b shows the Bleach treatment comparison between unshredded and shredded straw using
710 ml bleach diluted with 200L water. The AX strain was found to have a biological efficiency of
129.3% on shredded straw and 46.2% on unshredded straw, an 83.1% difference; while the ELM 1 strain
had an efficiency of 129.1% on shredded straw and 67.6% on unshredded straw, an 83.1% difference. The
PDJ strain had an efficiency of 84.2% on shredded straw and 25.1% on unshredded straw, a 59.1%
difference; while the PSAJ strain had an efficiency of 96.6% on shredded straw and 25.6% on unshredded
straw, a 71% difference.

Figure 1b: Bleach growth comparison

708mL/200L Bleach Comparison

140.00%

120.00%

100.00%

BE
80.00%
Shredded Straw
Regular Straw
60.00%

40.00%

20.00%

0.00%
AX ELM PDJ PSAJ
Strain

Figure 1b shows a direct comparison between the Bio efficiencies of shredded straw to regular unshredded straw for each of the representative
Pleurotus strains, AX, ELM, PDJ, and PSAJ using the 708 ml bleach to 200L of water sterilization method.

24
Figure 1c shows the Lime treatment comparison between unshredded and shredded straw using
355 g lime diluted with 200 L water. The AX strain was found to have an efficiency of 126.7%
on shredded straw and 33.2% on unshredded straw, a difference of 93.5%. The ELM 1 strain had
an efficiency of 168.9% on shredded straw and 53.2% on unshredded straw, a difference of
115.7%. The PDJ strain had an efficiency of 165.9% on shredded straw and 23.14% on
unshredded straw; a difference of 142.76%.The PSAJ strain had an efficiency of 117.1% on
shredded straw and 29.23% on unshredded straw, a difference of 87.87%.

Figure 1c: Lime growth comparison

Lime Growth Comparison

180.00%

160.00%

140.00%

BE 120.00%

100.00%
Shredded Straw
Regular Straw
80.00%

60.00%

40.00%

20.00%

0.00%
AX ELM PDJ PSAJ
Strain

Figure 1c shows a direct comparison between the Bio efficiencies of shredded straw to regular unshredded straw for each of the representative
Pleurotus strains, AX, ELM, PDJ, and PSAJ using the 355 ml lime to 200L of water sterilization method.

25
Figure 1d shows the Tide brand washing powder treatment comparison between the unshredded
and shredded straw using the 118.3 ml tide washing powder diluted with 200 L water. The AX
strain was found to have an efficiency of 134.7% on shredded straw and 79.5% on unshredded
straw; a difference of 55.2%. The ELM 1 strain had an efficiency of 128% on shredded straw
and 37.9% on unshredded straw; a difference of 90.1% . The PDJ strain had an efficiency of
83.2% on shredded straw and 23.13% on unshredded straw; a difference of 60.07%. The PSAJ
strain had an efficiency of 87% on shredded straw and 39.7% on unshredded straw; a difference
of 47.3%. As seen in graphs 1a through 1d, the biological efficiency for any of the test strains
using the shredded straw as a substrate gives a noticeably higher yield then using unshredded
straw as the substrate for all of the treatment methods.

Figure 1d: Washing powder growth comparison

Tide washing powder Growth Comparison

160.00%

140.00%

120.00%

100.00%
BE
Shredded Straw
80.00%
Regular Straw

60.00%

40.00%

20.00%

0.00%
AX ELM PDJ PSAJ
Strain

Figure 1d shows a direct comparison between the Bio efficiencies of shredded straw to regular unshredded straw for each of the representative
Pleurotus strains, AX, ELM, PDJ, and PSAJ using the 118.3 ml tide to 200L of water sterilization method.

26
Figure 2 shows the direct biological efficiency comparisons for shredded straw versus
unshredded straw for each of the four test strains used. As seen in this graph, none of the regular
unshredded straw strains for any of the treatment methods reached a biological efficiency higher
than 80% while none of the shredded straw strains reached a biological efficiency lower than
83.20%. It is interesting to note that the steam treatment method did not have the highest bio-
efficiency for all representative strains; which shows the potential for varying growth
requirements among this family of mushroom.

Figure 2: Unshredded straw vs. Shredded straw biological efficiency comparison graph
Shredded and Regular Straw Comparison Graph

200.00%

180.00%

160.00%

140.00%

120.00%
BE.
Shredded Straw
100.00%
Regular Straw

80.00%

60.00%

40.00%

20.00%

0.00%
AX ELM PDJ PSAJ AX ELM PDJ PSAJ AX ELM PDJ PSAJ AX ELM PDJ PSAJ

STEAM LIME 708mL/200L BLEACH Washing powder

Strain/Treatment

Figure 2 shows the direct comparison of Bio-efficiencies for each of the four Pleurotus strains grown using the four treatment methods for
shredded straw versus regular straw.

27
Figure 3 shows a direct comparison between the quantity of fruit bodies (fresh weight) picked in
grams for both the unshredded and shredded straw for each of the treatment methods Steam,
Lime, Bleach, and Washing Powder. This graph includes only representative Pleurotus strains
grown on both the shredded and unshredded substrate. It is interesting to note that the regular
straw gave higher yield average for ELM 1 treated with bleach and AX treated with washing
powder only, all the other strains treated with any of the treatment methods remained that the
shredded straw gave better yield in terms of fresh weight.

Figure 3: Regular straw vs. Shredded straw - fresh weight totals

Fresh Total Comparison

1200

1000

800
Fresh
Weight
SHRD STRW
600
REG STRAW

400

200

0
AX ELM PDJ PSAJ AX ELM PDJ PSAJ AX ELM PDJ PSAJ AX ELM PDJ PSAJ

STEAM LIME 708mL/200L BLEACH Washing powder

Strain/treatment

Figure 3 shows a direct comparison between the amount of fruit bodies (fresh weight) picked, in grams, for both the regular unshredded and
shredded straw for each of the treatment methods Steam, Lime, Bleach, and Tide.

28
Figure 4 shows the comparative all-strain average biological efficiencies for each treatment
method for both the regular unshredded and shredded straw substrates. The Steam treatment
averaged a biological efficiency of 132% on shredded straw and 63% on unshredded straw; the
bleach treatment averaged a biological efficiency of 110% on shredded straw and 73% on
unshredded straw; the lime treatment averaged a biological efficiency of 145% on shredded
straw and 69% on unshredded straw; and the washing powder treatment averaged a biological
efficiency of 108% on shredded straw and 90% on unshredded straw.

Figure 4: Regular straw vs. Shredded straw treatment averages

Treatment Comparison

160%

140%

120%

100%

BE.
Shredded
80%
Regular

60%

40%

20%

0%
Steam Avg Avg Lime Avg 708mL/200L Bleach Avg Washing powder
Treatment

Figure 4 shows the comparative all-strain average bio-efficiencies for each treatment method for both the regular unshredded and shredded straw
substraights.

29
Figure 5 shows the average biological efficiencies of both unshredded straw and shredded straw
substrates for each representative strain. The AX strain averaged a biological efficiency of 130%
on shredded straw and 62% on unshredded straw; ELM 1 averaged a biological efficiency of
150% on shredded straw and 67% on unshredded straw; PDJ strain averaged a biological
efficiency of 109% on shredded straw and 42% on unshredded straw; and the PSAJ strain
averaged a biological efficiency of 105% on shredded straw and 51% on unshredded straw.

Figure 5: Unshredded straw vs. shredded straw strain averages

Strain Avg Comparison

160%

140%

120%

100%

BE.
Shredded
80%
Regular

60%

40%

20%

0%
AX Avg Elm Avg PDJ Avg PSAJ Avg
Strain

Figure 5 shows the average bio-efficiencies of both regular unshredded straw and shredded straw substrates for each representative Pleurotus
strain.

30
Figure 6 shows representative photographs of the Pleurotus fruit bodies on the densely packed
shredded straw substraight using a variety of treatment methods; notice that the quantity of
fruitbody clusters on each of the bags are more numerous than the quantity of fruitbodies shown
in figure 7; this is due to the more densely packed shredded straw substrate. Picture 1 is a bag of
the AX strain treated with washing powder. Picture 2 is a bag of the PSAJ strain also treated with
the washing powder treatment method. Picture 3 shows bags of the PDJ and AX strains both
treated with washing powder. Picture 4 shows fruitbody clusters of an ELM 1 bag treated with
the steam treatment method.

Figure 6: Photos of shredded straw mushrooms

1. 2.

3. 4.
Figure 6 shows representative photographs of the Pleurotus fruit bodies on the densely packed shredded straw substrate using a variety of
treatment methods

31
 Figure 7 shows representative pictures of fruitbodies growing on unshredded straw treated with
the four representative treatment methods. The pictures in this figure can be compared to the
pictures in figure 6 in order to see the difference in growth between shredded and unshredded
straw when used as the substrate. Picture 1 shows the AX strain fruitbodies treated with washing
powder. Picture 2 shows the ELM 1 strain fruitbodies treated with steam. Picture 3 shows PDJ
strain fruitbodies treated with washing powder. Picture 4 shows the PSAJ strain fruitbodies
treated with lime.

Figure 7: Photos of unshredded straw mushrooms

1. 2.

3. 4.
Figure 7 shows representative pictures of fruitbodies growing on regular unshredded straw treated with the four representative treatment methods.

CONCLUSION:

Until now each farm cultivation operation has almost always been looked upon as a “stand
alone” operation, which is an operation complete in and of itself, whether it be for plants,
animals or fish. Raw materials come in, a crop is produced and the waste byproducts are
disposed of. It does not need to, and should not be like this. In fact, all of nature is a continuous
flow of building up followed by decay, with the residual waste from each step of the process
forming the raw material basis for the life forms to follow. Following this natural process, a
system of diversified agriculture has been developed and is presented where several successive
crops can be raised one after another, each crop depending upon the waste materials from the
previous step to act as the raw materials for the next step. In this fashion, several food crops can

32
be successfully grown, harvested, and used so that hunger and malnutrition need not be a factor
while the people of a country, city, town or village grow and develop.

33
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