Table of Contents

Cover

Title Page

List of Contributors

Acknowledgments

Preface

Chapter 1: Mushrooms and Human Civilization

1.1 Domestication of Mushrooms
References

Chapter 2: Current Overview of Mushroom Production in the World

2.1 Lentinula edodes
2.2 Pleurotus spp.
2.3 Auricularia spp.
2.4 Agaricus bisporus
2.5 Flammulina velutipes
2.6 Outlook
References

Chapter 3: Mushrooms: Biology and Life Cycle

3.1 Life Cycle of Fungi
3.2 The Subkingdom Dykaria
3.3 Homothallism, Heterothallism, and Amphithallism
3.4 Heterothallism
3.5 Homothallism
3.6 Amphithallism
3.7 Mating-Type Genes
3.8 Agaricus brasiliensis (Syn = A. subrufescens or A. blazei): An Intriguing Example of Amphithallism
3.9 Life Cycle of Uncultivated Mushrooms
3.10 The Truffles
3.11 Morels
3.12 The Chanterelles
3.13 The Matsutake
3.14 Porcini
3.15 Decreased Production of Mycorrhizal Mushrooms in the Northern Hemisphere
3.16 Fitness of Filamentous Fungi
3.17 Final Considerations
References

Chapter 4: Genetic Aspects and Strategies for Obtaining Hybrids

4.1 Agaricus bisporus
4.2 Oyster Mushroom (Pleurotus Species)
4.3 Conclusion
References

Chapter 5: Spawn Production

5.1 Our Spawn Industry Today
5.2 Basics
5.3 Spawn Production Techniques
5.4 Strain Selection
5.5 Strain Preservation and Degeneration
5.6 Production of Mother Cultures and Mother Spawn
5.7 Hygiene
5.8 Sterilization, Disinfection, and Filtration
5.9 Substrate Composition
5.10 Incubation
5.11 Conservation and Transport
References

Chapter 6: Compost as a Food Base for Agaricus bisporus

6.1 The Place of Agaricus strains in Nature
6.2 Compost Process Phase I
6.3 Preparing Raw Materials
6.4 Phase II
6.5 Phase III
References

Chapter 7: Casing Materials and Techniques in Agaricus bisporus Cultivation

7.1 General Aspects of Casing and Fruiting
7.2 Casing Materials
7.3 Casing Related Techniques
References

Chapter 8: The Bag or Block System of Agaricus Mushroom Growing

8.1 Overview of the System
8.2 Bags and Blocks in Use
8.3 Practical Use of the System – Phase I and Phase II
8.4 Practical Use of the System – Spawning and Phase III
8.5 Practical Use of the System – Casing through Cropping
References

Chapter 9: The Mushroom Industry in the Netherlands

References

Chapter 10: New Technology in Agaricus bisporus Cultivation

10.1 Introduction
10.2 Stages and Operations of the Production System
10.3 Conclusion
References

Chapter 11: Insect, Mite, and Nematode Pests of Commercial Mushroom Production

11.1 Fly Pests
11.2 Mite Pests
11.3 Nematode Pests
References

Chapter 12: Mushroom Diseases and Control

12.1 Introduction
12.2 Fungal Diseases
12.3 Bacterial Diseases
12.4 Viral Diseases
Further Reading

Chapter 13: Harvesting and Processing of Mushrooms

13.1 Introduction
13.2 Manual Harvesting
13.3 Mechanical Harvesting
13.4 Automatic Harvesting Systems
13.5 Washing Mushrooms
13.6 Canning Mushrooms
13.7 Conclusions
References

Chapter 14: Mushroom Farm Design and Technology of Cultivation

14.1 Selection of Site and Pre-Requisites
14.2 Components of a Mushroom Farm
14.3 Mushroom Cultivation Technology
14.4 Environmental Crop Management
14.5 Cultivated Edible Specialty Mushrooms
References

Chapter 15: Cultivation of Mushrooms in Plastic Bottles and Small Bags

15.1 Introduction
15.2 Characteristics of Mushroom Cultivation in Plastic Bottles and Small Bags
15.3 Cultivation Methods
15.4 Diseases in Mushroom Cultivation in Plastic Bottles and Small Bags
15.5 Pests in Mushroom Cultivation in Plastic Bottles and Small Bags
References

Chapter 16: Cultivation of Pleurotus ostreatus

16.1 Pleurotus Species as Edible Fungi
16.2 Pleurotus spp. as Recyclers
16.3 Cultivation of Pleurotus ostreatus
16.4 Diseases and Pests
16.5 Pleurotus spp. in Biotechnology
16.6 Future Challenges
References

Chapter 17: Characteristics, Production, and Marketing of the Sun Mushroom: The New Medicinal Cultivated Mushroom

17.1 Introduction
17.2 Morphological Characteristics
17.3 Spawn Production
17.4 Compost (Phase I and II)
17.5 Spawning and Mycelium Run
17.6 Casing Layer
17.7 Facilities used in the Production Process
17.8 Pinning and Harvest
17.9 Post-Harvest and Marketing
17.10 Medicinal and Chemical Characteristics of Mushrooms
17.11 Diseases and Pests
17.12 Spent Mushroom Substrate (SMS)
Acknowledgments
References

Chapter 18: Cultivation of Ganoderma lucidum

18.1 Introduction
18.2 Growing Conditions of Lingzhi
18.3 Cultivating Patterns
18.4 Production of the Substrate
18.5 Preparation of Spawn and Inoculation
18.6 Facilities
18.7 Genetic Breeding
18.8 Duration, Number of Flushes
18.9 Diseases and Pests in the Cultivation of Lingzhi
18.10 Medicinal Values
Acknowledgments
References

Chapter 19: Naturally Occurring Strains of Edible Mushrooms: A Source to Improve the Mushroom Industry

19.1 Edible Species and Their Cultivation
19.2 Steps for the Domestication of Naturally Occurring Species
19.3 Finding New Species for the Mushroom Production Industry: A Look Back at the Last Few Years
19.4 Conclusions
References

Chapter 20: Spent Mushroom Substrate Uses

20.1 Introduction
20.2 Characteristics of Spent Substrate
20.3 Bioremediation
20.4 Crop Production
20.5 Reuse in the Cultivation of Mushrooms
20.6 Food for Animals and Fish
20.7 Pest Management
20.8 Other Varied Uses
20.9 Conclusion
References

Chapter 21: Chemical, Nutritional, and Bioactive Potential of Mushrooms

21.1 Brief Introduction
21.2 Chemical Composition and Nutritional Properties
21.3 Bioactive Properties
21.4 Conclusions
References

Chapter 22: Medicinal Properties and Clinical Effects of Medicinal Mushrooms

22.1 Introduction
22.2 Current Perspectives and Advances
22.3 Medicinal Mushroom Drugs
22.4 Medicinal Mushroom Dietary Supplements
22.5 Evidences, Challenges, and Unsolved Problems
22.6 Medicinal Mushroom Natural Products as an Unclaimed Source for Drug Discovery
22.7 Unsolved Problems in the Study of Structural Characteristics, Isolation Process, Receptor-Mediated Mechanism and Antitumor Activity of MM β-Glucans
22.8 Medicinal Mushroom Clinical Studies
22.9 Conclusions
References

Index

End User License Agreement

List of Illustrations

Chapter 2: Current Overview of Mushroom Production in the World

Figure 2.1 Components (edible, medicinal, and wild) of the world mushroom industry based on percentage of total value ($63 billion) (2013).
Figure 2.2 World population (billions) versus world cultivated, edible mushroom production (billion kg).
Figure 2.3 Cultivated mushroom production in China and selected regions of the world, 2013 (billion kg).
Figure 2.4 World edible mushroom production (% of total) by genus (2013).
Figure 2.5 Mushroom production in China by genus (2013, CEFA 2014). Percentages following horizontal bars for each genus represent change from 2010 production levels (in billion kg).
Figure 2.6 Growth in world shiitake production (1980–2013; billion kg).
Figure 2.7 Sawdust-based “logs” of Lentinula edodes: (left) arranged in rows under shade cloth-covered shelters and (middle and right) with maturing mushrooms (photos: D. J. Royse and Q. Tan).
Figure 2.8 Percentage of total world Pleurotus spp. production in selected countries and regions.
Figure 2.9 Growth in world Auricularia spp. production (billion kg) (1986–2013).
Figure 2.10 Production of Agaricus bisporus in selected countries and regions (2013).
Figure 2.11 Growth in world production of Flammulina velutipes (billion kg) (1980–2013).

Chapter 3: Mushrooms: Biology and Life Cycle

Figure 3.1 Scanning electron micrographs of Agaricus brasiliensis gills. A–C: Bisporic basidia. D–F: Trisporic basidia. G–1: Tetrasporic basidia. Bar = 1 mm. Herreira et al. (2012), Mycologia. With permission.
Figure 3.2 Scanning electron micrographs of Agaricus brasiliensis gills. The arrow indicates a basidiospore linked to a connection hyphae and two basidia. Arrowheads indicate sterigmata connected to one another by a hypha. Bar = 1 mm.
Figure 3.3 Theoretical life cycle of the genus Morchella. Dotted lines indicate possible routes. AlvaradoCastillo, G.; Mata, G.; Sangabriel-Conde, W. Understanding the life cycle of morels (Morchella spp.). Revista Mexicana de Micologia, v. 40, p. 47-50, 2014. With pennission.

Chapter 4: Genetic Aspects and Strategies for Obtaining Hybrids

Figure 4.1 Global mushroom production (based on data from FAOSTAT, 2011).
Figure 4.2 Global mushroom production over the last four decades.
Figure 4.3 Three factors impeding hybridization in button mushrooms.
Figure 4.4 Life cycle of Agaricus bisporus var. bisporus.
Figure 4.5 Crop of DMR-03, a single spore isolate, at a commercial unit. (See color plate section for the color representation of this figure.)
Figure 4.6 Steps in development of U1 and U3.
Figure 4.7 Crop of NBS-1 (left), cross section of fruiting body (top right), and quality of fruiting bodies after storage at room temperature for 48 h (bottom right). (See color plate section for the color representation of this figure.)
Figure 4.8 Crop of NBS-5 (left), cross section of fruiting body (top right), and quality of fruiting bodies after storage at room temperature for 48 h (bottom right). (See color plate section for the color representation of this figure.)
Figure 4.9 Variability in stipe, pileus, gills and gill cavity in different accessions of button mushroom. (See color plate section for the color representation of this figure.)
Figure 4.10 Variation in browning after 2 h. (See color plate section for the color representation of this figure.)
Figure 4.11 Dendrogram separating fertile and non-fertile isolates.
Figure 4.12 RAPD of fertile and non-fertile isolates of A. bisporus.
Figure 4.14 ISSR profile of five fertile and five non-fertile isolates using P-3 and P-8 primers.
Figure 4.13 Diversity in homokaryons and heterokaryons.
Figure 4.15 Intermating of non-fertile isolates.
Figure 4.16 Five possible matings on mixing homokaryotic mycelium with spore print of other parental strain.
Figure 4.17 Above: yield and downward linear growth of non-fertile (left) and fertile (right) isolates, and Below: scatter plot of yield versus downward linear growth of 69 SSIs of A. bisporus.
Figure 4.18 Naturally occurring Pleurotus species in forests of Maharashtra (India).
Figure 4.19 Analysis of ITS2 sequences in Pleurotus species available at DMR Solan.

Chapter 5: Spawn Production

Figure 5.1 The mushroom production pyramid.
Figure 5.2 Example of inoculation practice of small grain bags without inbuilt filter.
Figure 5.3 Evolution in breathing bag technology.
Figure 5.4 Comparison of product flow of two spawn production methods: sterilization in bulk or individual bags.
Figure 5.5 Manual inoculation of large grain spawn bags in front of laminar flow. Note the anemometer measuring the horizontal LAF’s air velocity at the top and the sealing machine at the right.
Figure 5.6 Bag tumbler.
Figure 5.7 Example of a bulk vessel for the production of Agaricus grain spawn, inoculated with liquid mother spawn.
Figure 5.8 Lag time of liquid spawn vs. grain spawn of Pleurotus eryngii. If the amount of liquid inoculum is higher, the lag times are noticeably shorter
Figure 5.9 Keeping cryovials in a deep freezer.
Figure 5.10 Keeping cultures in a culture collection.
Figure 5.11 Double-packing cultures in fridges limits risks of cross-contamination.
Figure 5.12 Inoculating a mother culture petri dish with mushroom tissue.
Figure 5.13 Making PA (potato agar) and sterilizing slants.
Figure 5.14 Subculturing and the creation of four lines. Example of a four-step breeding program. Multiplication 1: subculturing = creation of five mother cultures (1 MCT and 4 MCP). One of these will be kept as backup, the other four become a line parent and are meant for production purposes. Multiplication 2: expansion from MCP to eight small mother spawn bags “A” (MSBA). Multiplication 3: expansion from one mother spawn bag “A” (MSBA) to eight large mother spawn bags “B” (MSBB). Multiplication 4: expansion from one mother spawn bag “B” (MSBB) to 24 spawn bags (SB).
Figure 5.15 Making pre-culture spawn in cleanroom conditions.
Figure 5.16 Floor plan of a grain spawn lab, depicting overpressure flow and pressure fall. Note the size of the mother spawn department compared to the spawn department.
Figure 5.17 Daily cleaning in the spawn cleanroom department.
Figure 5.18 Compression of gases in bag during autoclaving.
Figure 5.19 Gravity cycle – comparison of different spawn production systems.
Figure 5.20 Influence of gases on sterilization temperature.
Figure 5.21 Vacuum sterilization cycle – sterilization of porous and bagged goods.
Figure 5.22 Flow of aseptic overpressure air in cooling room.
Figure 5.23 Damaged HEPA filters in need of replacement.
Figure 5.24 Millet is an example of a grain type which has a good particle size.
Figure 5.25 Incubating mother culture displaying various zones of incubation.
Figure 5.26 Relative increase of C0₂ concentration in four spawn bag types in function of distance to headspace at 100% colonization. Head, high, mid, and low indicate the measuring levels inside the bag. The SD bag has filters evenly spread over the whole surface of the bag, the BK, UB, and UT bags have only a top filter strip or patch (Van Nuffel et al., 2016).

Chapter 6: Compost as a Food Base for Agaricus bisporus

Figure 6.1 After a good phase II process, there are a lot of Actinomycetes that can be seen as white spots on the compost.
Figure 6.2 Bale dunking to get a uniform wet start.
Figure 6.3 Left: Chicken manure cutter for high volumes. Right: Wetting process for corncobs.
Figure 6.4 In the US, many compost farms use Pannell mixing machines.
Figure 6.5 A relatively cheap manure mixing wagon.
Figure 6.6 Open air pile composting.
Figure 6.7 (a) An aerated bunker floor; (b) Modern bunker fermentation; and (c) Overhead filling system for filling bunkers as uniformly as possible.
Figure 6.8 Compost hall indoor facility.
Figure 6.9 Softening of straw after wetting on aerated floors.
Figure 6.10 Computer control is a must in modern compost facilities and is used in phase I and II of composting.
Figure 6.11 Ammonia pump.
Figure 6.12 One-zone system where all stages of the process happen in the same room.
Figure 6.13 Aerated floor from an open tunnel floor.
Figure 6.14 Left: Filling cassette for phase II tunnels. Right: Pulling winch for emptying compost from open tunnel floors with nylon nets.
Figure 6.15 Incubation in tunnels that can fill with phase III compost and casing together.
Figure 6.16 Filtering overpressure during spawning is necessary to protect the compost during spawning against infections. The plastic air ducts at the top distribute the clean air throughout the whole spawning hall.
Figure 6.17 (System under phase II). There are different methods of incubation. In plastic bags, blocks, and even in modern shelves; all of them need to be cased later.

Chapter 7: Casing Materials and Techniques in Agaricus bisporus Cultivation

Figure 7.1 Fruiting of mushrooms on a peat-based casing (interval A–B: 5 days; interval B–C: 2 days).
Figure 7.2 Compost (lower) and casing (upper) layers in mushroom production.
Figure 7.3 Hand-operated ruffling apparatus.
Figure 7.4 Detail of a ruffling machine: rod with teeth and pressure roller.

Chapter 8: The Bag or Block System of Agaricus Mushroom Growing

Figure 8.1 Superior quality of bag system with a single layer of bags on the floor.
Figure 8.2 Bagging machine.
Figure 8.3 Example of blocking machine platform, each separation refers to the block size.
Figure 8.4 Spawned blocks (palletizing).
Figure 8.5 Bags closed (not hermetically) for the spawn run.
Figure 8.6 Mechanical casing of blocks winched onto shelves.
Figure 8.7 Dedicated incubation room with tight bag/block placement, but less than optimum heat transfer.
Figure 8.8 Dedicated incubation room for bags (a) and blocks (b).
Figure 8.9 Mechanical filling of the shelves.
Figure 8.10 Casing machine.
Figure 8.11 Individual “trays” with blocks.
Figure 8.12 Blocks with plastic cut away for casing.
Figure 8.13 Mechanized casing.
Figure 8.14 Heavy casing which limits mycelial colonization (approximately 4 cm).
Figure 8.15 Equipment used for irrigation of the casing layer, called the “water tree.”
Figure 8.16 Mushrooms accessible for harvest on bags (a) and blocks (b).
Figure 8.17 Individual “trains”, an efficient way of disposing of blocks.
Figure 8.18 Harvested mushrooms.

Chapter 9: The Mushroom Industry in the Netherlands

Figure 9.1 Using compost blocks in shelves.
Figure 9.2 The huge roof construction over phase I compost piles for many years. In the same place now are modern tunnels for phase II and III.
Figure 9.3 Many years before the development of tunnels, the CNC delivered phase I compost to all members in the Netherlands. The CNC filled these as well and many years on, they empty the rooms too.
Figure 9.4 Head filling machine for filling phase III compost and casing together in modern Dutch mushroom farms.
Figure 9.5 Automatic lorry: the yellow cable follows after picking to connect the batteries.
Figure 9.6 Indoor compost facility of the CNC. The CNC also has another location containing many tunnels. The plan is to develop an indoor facility there as well.
Figure 9.7 Typical Dutch mushroom farm in 2016.
Figure 9.8 In the filling hall you cannot see compost conveyer belts because the input belts are above the ceiling and the output belts below the floor. The tunnels are filled completely automatically, with no person in the tunnel. There is only one laborer watching a monitor to control the filling process.
Figure 9.9 An actual situation in the Netherlands where they fill phase III compost plus supplement and casing using modern equipment all together on a typical modern Dutch mushroom farm. In roughly 1 hour they fill about 200–300 m² with these materials. The organization is so good that the head filling machine (from a special filling company) plus the truck with compost and the truck with casing soil, from different companies, arrive at the same time. Just 90 minutes later, everything is gone and the room is filled perfectly.

Chapter 10: New Technology in Agaricus bisporus Cultivation

Figure 10.1 Plan view of a typical mushroom farm.
Figure 10.2 Modern composting plant in La Rioja (Spain).
Figure 10.3 Filling tray with compost and casing on plastic film.
Figure 10.4 Line filling the trays.
Figure 10.5 Detail of irrigation (indicated by the pale arrow) and ruffling (indicated by the dark arrow).
Figure 10.6 Line of casing, conveyor belt, and mixing drum.
Figure 10.7 Facility used for the new technology of A. bisporus cultivation. Incubation and crop rooms with the line of harvesting.
Figure 10.8 Different views of the harvest line.

Chapter 11: Insect, Mite, and Nematode Pests of Commercial Mushroom Production

Figure 11.1 Sciarid flies, adult.
Figure 11.2 Cecid larvae, orange cecid larvae on a mushroom.
Figure 11.3 Sciarid larvae with distinctive black head capsule.
Figure 11.4 Howardula husseyi, a macerated female phorid fly showing Howardula husseyi larve, gravid female, and eggs.
Figure 11.5 Red pepper mites on the surface of mushrooms.
Figure 11.6 Phorid larvae.

Chapter 12: Mushroom Diseases and Control

Figure 12.1 Undifferentiated masses of mushroom tissue as a result of infection by Lecanicillium fungicola.
Figure 12.2 Imperfectly formed mushroom with distorted stalk caused by Lecanicillium fungicola.
Figure 12.3 Cap spotting of Agaricus bisporus caused by Lecanicillium fungicola.
Figure 12.4 Undifferentiated tissue mass of Agaricus bisporus affected by Mycogone perniciosa.
Figure 12.5 Mycogone perniciosa on the surface of developed mushrooms. Amber droplets of liquid can be seen on the diseased mushrooms.
Figure 12.6 Cobweb mycelium (Cladobotryum mycophilum) on the surface of the casing and attacking mushrooms (Agaricus bisporus).
Figure 12.7 Cobweb mycelium (Cladobotryum mycophilum) causing cap spotting.
Figure 12.8 Brown spots on a mushroom cap, caused by Trichoderma aggressivum.
Figure 12.9 Trichoderma aggressivum colonizing compost.
Figure 12.10 Trichoderma mold colonizing casing.
Figure 12.11 Mycelium and ascocarps of Diehliomyces microsporus on the casing surface.
Figure 12.12 Dense mycelium of Diehliomyces microsporus on the compost.
Figure 12.13 White plaster mold on the compost surface.
Figure 12.14 Brown plaster mold on the casing surface.
Figure 12.15 Bacterial blotch causing cap spotting.
Figure 12.16 Close up view of bacterial blotch causing cap spotting.
Figure 12.17 Internal stipe necrosis caused by Ewingella americana.
Figure 12.18 Internal stipe necrosis. Mushroom with a hollow center.
Figure 12.19 Mushrooms affected by La France disease (drumstick symptom).

Chapter 14: Mushroom Farm Design and Technology of Cultivation

Figure 14.1 General layout of 250 TPA (tons per annum) mushroom farm.
Figure 14.2 Compost yard, bulk chamber, and spawning area.
Figure 14.3 Cross section of a compost bulk pasteurization chamber (length 13 m/45’, width 3 m/10’, height 4 m/13’).
Figure 14.4 Casing pasteurization chamber.
Figure 14.5 Layout plan of a spawn lab.
Figure 14.6 Front cross-sectional view of cropping rooms.
Figure 14.7 Low cost growing room.
Figure 14.8 Internal layout of a cropping room (length 18 m/60’, width 7 m/22’, height 4.4 m/15’).
Figure 14.9 Gray oyster mushroom (a), black oyster mushroom (b), and white oyster mushroom (c).
Figure 14.10 Pink oyster mushroom (a) and yellow oyster mushroom (b).
Figure 14.11 King oyster mushroom.
Figure 14.12 Crop of black ear mushroom.
Figure 14.13 Buna shimeji mushroom.
Figure 14.14 Milky mushroom (a) and paddy straw mushroom (b).
Figure 14.15 Cultivation of portobello brown (a) and portobello with button mushroom, note the difference in color and size (b).

Chapter 15: Cultivation of Mushrooms in Plastic Bottles and Small Bags

Figure 15.1 Small-scale bag cultivation of Auricularia in Vietnam.
Figure 15.2 Large facility for bag cultivation of Grifola frondosa in Japan.
Figure 15.3 Polypropylene bottles and tray for cultivation.
Figure 15.4 Polypropylene small bags with and without microporous filter.
Figure 15.5 Mixing substrate.
Figure 15.6 Automatic filling machine for bottle cultivation.
Figure 15.7 Filling the substrate for small-scale bag cultivation of Shiitake in China.
Figure 15.8 Large autoclaves for sterilization of substrate.
Figure 15.9 Automatic inoculation machine for bottle cultivation.
Figure 15.10 Stacking trays containing 25 bottles in incubation room.
Figure 15.11 Scratching machine for fruiting.
Figure 15.12 Automatic harvesting machine in H. marmoreus cultivation.
Figure 15.13 Fruit body development of F. velutipes after scratching.
Figure 15.14 F. velutipes fruiting in bottle cultivation.
Figure 15.15 F. velutipes fruiting in small bag cultivation in China.
Figure 15.16 Fruit body development of H. marmoreus after scratching.
Figure 15.17 H. marmoreus fruiting in bottle cultivation.
Figure 15.18 P. eryngii fruiting in bottle cultivation under LED illumination.
Figure 15.19 Packaging of P. eryngii.
Figure 15.20 P. nameko fruiting in bottle cultivation.
Figure 15.21 P. ostreatus fruiting in bottle cultivation.
Figure 15.22 G. frondosa fruiting on colonized substrate in bag cultivation.
Figure 15.23 L. edodes fruiting on colonized substrate removed from bags
Figure 15.24 Fruit bodies of Flammulina velutipes affected by Cladobotryum varium. Mushrooms covered with the fluffy mycelium in the early stage (a) and enveloped with soft powdery mycelium at a later stage of disease development (b).
Figure 15.25 C. varium attacking fruit bodies of Hypsizygus marmoreus.
Figure 15.26 Antagonistic contaminants competing with mushroom mycelium. The diamond-shaped patches show that the contaminants were introduced into the bottles during the cooling of substrate or inoculation.
Figure 15.27 Bacterial contamination in the substrate (right). Mushroom mycelial growth is strongly prevented by preceding fully colonized bacteria.
Figure 15.28 Patch-like blue-green colonies of Penicillium caused by an invasion of the mites fed the spores. Patch-like contamination patterns show the moving trace of mites in the substrate.

Chapter 16: Cultivation of Pleurotus ostreatus

Figure 16.1 (a) Cluster of Pleurotus ostreatus basidiomata. (b) Pleurotus cornucopiae var. “citrino-pileatus,” the golden oyster mushroom. (c) Pleurotus eryngii growing on cottonseed hulls substrate.
Figure 16.2 Mushroom mycelium growing on potato dextrose agar (left), partially colonized grain (center) and completely colonized grain (right). Reproduced with permission from the Spawn Laboratory, The Pennsylvania State University.
Figure 16.3 (a) Liquid spawn incubation system in China. (b) Oyster cultivation in small bags inoculated with liquid spawn (China).
Figure 16.4 (a) Chopped wheat straw. (b) Pelletized cottonseed hulls. (c) Tractor about to unload homogenized wheat straw substrate into a pasteurization tunnel. (d) Washed and disinfected tractor used to transfer the substrate from the pasteurization tunnel to the spawning room.
Figure 16.5 (a) Substrate incubation in black polypropylene bags with slits and (b) holes.
Figure 16.6 Oyster primordia developing from a fully colonized substrate bag.
Figure 16.7 (a) Heavy cluster of oyster mushrooms' primordia. (b) Young oyster mushrooms growing on cottonseed hulls/wheat straw substrate. (c) Mature oyster mushrooms growing on cottonseed hulls/wheat straw substrate.
Figure 16.8 (a) Harvested golden oyster (P. cornucopiae var. “citrino-pileatus”) mushrooms and (b) pink oyster mushrooms (P. djamor var. salmoneostramineus) in plastic crates.
Figure 16.9 P. tolaasii growing on young P. eryngii basidiomata.
Figure 16.10 P. tolaasii lesions on P. eryngii stipes are characterized by striations.
Figure 16.11 (a) Scarid fly, adult fly and larva. (b) Phorid fly. (c) Cecid fly, adult and larvae. Source: Reproduced with permission of The Pennsylvania State University.

Chapter 17: Characteristics, Production, and Marketing of the Sun Mushroom: The New Medicinal Cultivated Mushroom

Figure 17.1 (a) Morphological aspects of the mushrooms, (b) spore print, and (c) elliptical spore.
Figure 17.2 On the left, a machine used to turn the compost (semi-mechanized, it still needs some manual labor); right, manual turning made in smaller, family farms.
Figure 17.3 On the left, disposal of compost on wooden shelves for mycelia growth; right, detail of the folded bags that allow the application of insecticide.
Figure 17.4 On the left, compost colonized by Sun mushrooms; right, compost colonized by A. bisporus.
Figure 17.9 Yield of the Sun mushroom depending on the strain, compost, the casing layer, environmental conditions, and type of irrigation (data provided by Zied and colleagues from their experiences during 2005–2009).
Figure 17.5 Facilities used in the production process: (a) greenhouses; (b) aligned “Tetra Pak” barracks; (c) outdoor; (d) controlled chambers; and (e) rustic structures in forests.
Figure 17.6 Scheme of 120 days production, under controlled conditions (climate chambers), and semi-controlled (greenhouses and aligned barracks), depending on the temperature of the compost, the air (°C), and relative humidity (%).
Figure 17.7 On the left, mushrooms with a natural color on the surface of the pileus; right, mushrooms with the “burned” pileus, as indicated by the arrows.
Figure 17.8 Temperatures of growing cycle in the fast cooling method (five flushes).
Figure 17.10 Diagram of process of post-harvest: (a) mushrooms picked; (b) washing; (c) clean mushrooms; (d) cutting; (e) dehydrator; and (f) final product for export.
Figure 17.11 Dehydrated mushrooms, light colored (left) packages containing 2 kg of dehydrated mushrooms ready for export (center); and mushrooms of a darker shade that have not passed the antioxidant treatment (right).
Figure 17.12 States of development of mushrooms and their current market values (dried), * FP–non-standard (crushing of these mushrooms is recommended to give value to the product).
Figure 17.13 Scatter chart scores. Mushroom divided according to their parts and degree of maturation: ◦ - Closed/Pileus; • - Closed/Stipe; ▵ - Open/Pileus; ▴ - Open/Stipe; ▫ - Closed mushroom/Pileus + Stipe; ▪ - Open mushroom/Pileus + Stipe. EV2 – Energy value; M: Moisture; HE: Hemicellulose; A: Ash; N/P: Nitrogen and protein; ADF: Acid detergent fiber; CE: Cellulose; CF: Crude fiber; TC: Total carbohydrates.
Figure 17.14 (a) Deformation due to the presence of L. fungicola (globose aspect of mushroom), (b) presence of bacteria of the genus Pseudomonas, and (c) internal view of mushroom infected by Mycogone (wet mass).
Figure 17.15 (a) Mushroom infected with the mycoparasite S. megalocarpus, where it is possible to see its mycelium on lamellae of the Sun mushroom; (b) mushrooms presenting cracks on stipe and brown coloration in the pileus; (c) mycelium of S. megalocarpus in the casing layer.
Figure 17.16 (a) Flies in the mushroom, (b) light trap entomological impregnated tail, and (c) close view of the trap with dead flies.

Chapter 18: Cultivation of Ganoderma lucidum

Figure 18.1 Several Ganoderma species approved for use in production of health products. (a) G. sinensis; (b) G. tsuage; (c) G. lucidum.
Figure 18.2 Cultivating pattern of Lingzhi for producing fruiting bodies.
Figure 18.3 Cultivating patterns of Lingzhi for producing mycelial biomass.
Figure 18.4 Schematic diagram of STRs. 1. Charging hole; 2. Mixer; 3. Cooling water inlet; 4. Clean hole; 5. Electric engine; 6. A device for measurement and control; 7. Blow vent; 8. Cooling water outlet; 9. Liquid-state medium; 10. Sterile air.
Figure 18.5 Display mode of the incubated bags for substitute cultivation.
Figure 18.6 Schematic diagram of development process of Lingzhi fruiting body. (a) Bud-breaking stage; (b) Bud-developing stage; (c) Developing stage; (d) Growth stage; (e) Maturity stage.

Chapter 19: Naturally Occurring Strains of Edible Mushrooms: A Source to Improve the Mushroom Industry

Figure 19.1 Production of Agaricus pseudoargentinus in A. bisporus compost.
Figure 19.2 Production of Lentinus tigrinus on sawdust. Detail of pileus (a) and massive primordia formation (b).
Figure 19.3 Production of Polyporus tenuiculus in bags.
Figure 19.4 Production of Polyporus tenuiculus in logs.
Figure 19.5 Production of Agrocybe cylindracea in supplemented wheat straw.
Figure 19.6 Production of Pleurotus albidus on supplemented sawdust.
Figure 19.7 Production of Gymnopilus pampeanus in bags.

Chapter 22: Medicinal Properties and Clinical Effects of Medicinal Mushrooms

Figure 22.1 Objectives of clinical trials for herbal and mushroom medicines.

List of Tables

Chapter 4: Genetic Aspects and Strategies for Obtaining Hybrids

Table 4.1 Variation in type of basidia in two varieties of Agaricus bisporus (button mushroom).

Chapter 7: Casing Materials and Techniques in Agaricus bisporus Cultivation

Table 7.1 Interrelated factors involved in mushroom fruiting.
Table 7.2 Main parameters and methods used in the analysis of casing soils and their components.

Chapter 10: New Technology in Agaricus bisporus Cultivation

Table 10.1 Sample of managing environmental conditions for fruiting of A. bisporus for the fresh market.

Chapter 14: Mushroom Farm Design and Technology of Cultivation

Table 14.1 Edible specialty mushrooms/optimum temperature for cultivation.

Chapter 16: Cultivation of Pleurotus ostreatus

Table 16.1 Species of Pleurotus spp. commercially or experimentally cultivated around the world.
Table 16.2 Nutritional composition (fat, carbohydrates, dietary fiber, and protein) of different edible mushrooms (numbers are percent of the fresh weight).

Chapter 17: Characteristics, Production, and Marketing of the Sun Mushroom: The New Medicinal Cultivated Mushroom

Table 17.1 Yield (kg mushroom/12 kg of compost) obtained for five strains of Sun mushroom in three different composts.
Table 17.2 Formulation of Sun mushroom compost, for the final production of 15 tons; first and second formulation “without manure,” and third formulation classic compost.
Table 17.3 Operations to be carried out during composting (first formulation).
Table 17.4 Values of yield (kg of mushrooms/12 kg of compost); unit weight of mushrooms (yield divided by the number of mushroom), and number of mushrooms (collected in 12 kg of compost).
Table 17.5 Recommended environmental conditions for the cultivation of Sun mushrooms after casing application.
Table 17.6 Price of the most commonly cultivated mushroom species in different countries, comparing them to the commercial price of Sun mushrooms in Brazil.
Table 17.7 Mycochemicals characteristics of mushrooms depending on physiological and morphologic characteristics.

Chapter 21: Chemical, Nutritional, and Bioactive Potential of Mushrooms

Table 21.1 Approximate composition and energy values of wild and cultivated mushroom species.
Table 21.2 Main fatty acids (relative percentages) found in wild and cultivated mushroom species.
Table 21.3 Main sugars (g/100 g dw) found in wild and cultivated mushroom species.
Table 21.4 Trace metals (µg/g dw) found in wild and cultivated mushroom species.
Table 21.5 Free amino acids (g/100 g dw) found in wild and cultivated mushroom species.
Table 21.6 Antioxidant activity of mushroom species.
Table 21.7 Antimicrobial activity of mushroom species.
Table 21.8 Antitumor activity of mushroom species.

Chapter 22: Medicinal Properties and Clinical Effects of Medicinal Mushrooms

Table 22.1 Levels of evidence.
Table 22.2 Therapeutic activities of medicinal mushroom compounds and extracts evaluated in clinical studies.

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List of Contributors

Edgardo Albertó
Laboratory of Mycology and Mushroom Cultivation
Instituto de Investigaciones Biotecnológicas-
Instituto Tecnológico Chascomús (UNSAM-CONICET)
Buenos AiresArgentina

Johan Baars
Wageningen UR, Plant Breeding WageningenNetherlands

Jos Buth
Viña del Mar
Region VChile

Ângela Fernandes
Centro de Investigação de Montanha (CIMO)
ESA, Instituto Politécnico de Bragança
BragançaPortugal

Isabel C.F.R. Ferreira
Centro de Investigação de Montanha (CIMO)
ESA, Instituto Politécnico de Bragança
BragançaPortugal

Francisco J. Gea
Centro de Investigación
Experimentación y Servicios del Champiñón (CIES)
Quintanar del Rey (Cuenca)Spain

Arcadio Gómez
Mushiberica Consultores
AlbaceteSpain

Sandrina A. Heleno
Centro de Investigação de Montanha (CIMO)
ESA, Instituto Politécnico de Bragança
BragançaPortugal

Behari Lal Dhar
NNMushroom Consulting India/ICAR-Directorate of Mushroom Research SolanIndia

Shwet Kamal
ICAR-Directorate of Mushroom Research, SolanIndia

Kasper Moreaux
Mycelia, Spawn Production and School for Professionals in the Mycelium IndustryNeveleBelgium

María J. Navarro
Centro de Investigación
Experimentación y Servicios del Champiñón (CIES)
Quintanar del Rey (Cuenca)Spain

José Emilio Pardo González
Escuela Técnica Superior de Ingenieros Agrónomos y de Montes (ETSIAM)
Universidad de Castilla-La ManchaAlbaceteSpain

Arturo Pardo-Giménez
Centro de Investigación
Experimentación y Servicios del Champiñón (CIES)
Quintanar del Rey (Cuenca)Spain

John Pecchia
Plant Pathology and Environmental Microbiology
Penn State University
University Park, PAUSA

Danny Lee Rinker
University of Guelph
Guelph, ONCanada

Manuela Rocha de Brito
Department of Biology, University of Lavras (UFLA)Brazil

Alma E. Rodriguez Estrada
Biology Department
Aurora University
Aurora, ILUSA

Daniel J. Royse
Department of Plant Pathology and Environmental Microbiology, The Pennsylvania State University, University Park, PAUSA

Raymond Samp
Agari-Culture Consulting Services
San MarcosTexasUSA

Manjit Singh
ICAR-Directorate of Mushroom Research, SolanIndia

Eustáquio Souza Dias
Department of Biology, University of Lavras (UFLA)Brazil

Qi Tan
Shanghai Academy of Agricultural Sciences ShanghaiChina

Juan Valverde
Food Research and Technology Programme
Research and Development Department
Monaghan Mushrooms
MonaghanIreland

Solomon P. Wasser
Institute of Evolution and Department of Evolutionary and Environmental Biology
Faculty of Natural Sciences
University of Haifa, Haifa, Israel
and N.G. Kholodny Institute of Botany
National Academy of Sciences of Ukraine
KievUkraine

Katsuji Yamanaka
Director
Kyoto Mycological Institute
KyotoJapan

Xuan-Wei Zhou
School of Agriculture and Biology
Engineering Research Center of Cell & Therapeutic Antibody (Ministry of Education)
Shanghai Jiao Tong University
ShanghaiPeople’s Republic of China

Diego Cunha Zied
Universidade Estadual Paulista (UNESP)
São PauloBrazil

Preface

The term Mushrooming, or mushroom cultivation, refers to the intentional and directed production of mushrooms as a substitute for wild collection in fields and forests with a harvest under defined conditions of growing, resulting in strict quality control and food safety without risk of consumption of poisonous or toxic species, and with guaranteed benefits from fungi.

Although knowledge about the cultivation of edible and medicinal mushrooms is practically the same throughout the world, there are significant differences between countries and even within the same country. These are primarily associated with different socioeconomic conditions. In this way, just as there are large-scale growers, other smaller-scale plants act as a complement to the family economy, while very basic and rustic facilities coexist with others that operate on a high technological level.

This book involves a multidisciplinary approach that includes aspects of agriculture and agronomy, microbiology, biology, biotechnology, chemistry, environmental management, food technology, and health, among others. With a global and collaborative purpose, the book consists of 22 chapters written by 28 authors, from 15 different countries, who are recognized experts in the different areas that compose this activity. We thank them all for their participation.

The different areas of the science of cultivation are approached, so the book can serve as a tool for researchers, professors, technical specialists, and growers, and as an introduction for both students and anyone interested in the world of mushrooming knowledge as a business opportunity or out of simple curiosity.

Diego Cunha Zied, Ph.D.
Professor and Head of Centro de Estudos em Cogumelos
Faculdade de Ciências Agrárias e Tecnológicas
Universidade Estadual Paulista (UNESP – Campus de Dracena)
Brazil

Arturo Pardo-Giménez, Ph.D.
Researcher of Centro de Investigación, Experimentación y Servicios del Champiñón
Patronato de Desarrollo Provincial, Diputación Provincial de Cuenca
Spain

Edible and Medicinal Mushrooms

Technology and Applications

List of Contributors

Acknowledgments

Preface