ANTIMYCOTIC

PPROPERTY OF PEARL
OYSTER MUSHROOMS
Pleurotus ostreatus TESTED
ON SWINE

Reymond Gabriel L. Abedania

John Carlo G. Baribal
Laizel Mareli L. De Castro
Researchers

Mr. Arturo S. Blanco

Research Adviser

iii
Antimycotic Property of Pearl Oyster Mushrooms

Pleurotus ostreatus Tested on Swine

A Thesis
Presented to the Faculty of
SAN FABIAN NATIONAL HIGH SCHOOL
Special Science Class
S.Y. 2019-2020

In Partial Fulfillment of the

Requirements in Research I

Reymond Gabriel L. Abedania

John Carlo G. Baribal
Laizel Mareli L. De Castro
Grade 9 Special Science Class – Gamma Cassiopeiae

Mr. Arturo S. Blanco

Research Adviser

March 2020

ii
 Acknowledgement

The researchers would like to extend their heartfelt gratitude, first, to God who

helped the researchers through making the research. He gave the researchers the

patience, wisdom, and knowledge on upon conducting the research.

Furthermore, the researchers would like to thank the San Fabian National High

School for providing additional knowledge about the research – specifically, the school’s

library;

The Science and Technology Department under the leadership of Ms.

Luzviminda S. Juguilon for inspiring the researchers in completing the research;

To all the San Fabian National High School’s Departments, department heads,

and teachers for giving additional information about research and technological

advancements as stated so;

The researchers also want to take this opportunity to express gratitude to Mr.

Arturo S. Blanco for monitoring and constant encouragement throughout the course of

this research.

The researchers also want to thank the San Fabian hog-growers for letting the

researchers use their swine on upon conducting the scientific research and the

mushroom producers in Mapandan, Pangasinan who supplied the researchers needs in

terms of the researcher’s variable.

The researchers are thankful for their fellow researchers who continually

encouraged and supported them in whatever the researchers faced.

iii
 Lastly, the researchers also deeply honor their love and sincerest thanks to their

family members who supported them from the beginning of the research after the end of

it and for giving them strength in times of difficulty.

iv
 Dedication

The researchers dedicate this research study to Almighty God who guided the

researchers from the very beginning up to the end of conducting the research.

They would also like to dedicate the study to their parents who supported them

on upon conducting the research study.

They also dedicate the research study to the Philippines’ Department of

Agriculture to better sustain the quality of life in the Philippines, the researchers’

homeland.

Lastly, the researchers dedicate the research study to future researchers who

would like to develop further studies about the antimycotic properties of pearl oyster

mushroom Pleurotous ostreatus

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 ABSTRACT

Pearl oyster mushrooms are abundant in Pangasinan. They grow on dead trees

and/or cultured by farmers to get extra income from their jobs. Many are forgetting the

benefits that they can get in pearl oyster mushrooms which the researchers had an idea

to benefit humans and animals alike. Pork has been used for centuries as food. It

supplies energy that is needed for humans. Humans solved pig’s illnesses, yet,

outbreaks are still widespread. When there is an outbreak, the price of pork will increase

due to lack of resources. The results don’t just affect the producers, but also the

consumers. The researchers confirmed the antimycotic ability (property) of pearl oyster

mushrooms which are ought to kill Candida spp. and ringworms of different types. High

(peptides and proteins) and low (sesquiterpenes, terpenes, steroids, organic acids, and

quinolones) molecular substances from pearl oyster mushrooms are seen by the

researchers on upon conducting a confirmatory test – phytochemical analysis – in

relation to reviews of related literature and studies. Tested swine resulted to a

confirmation of the average time of healing of pearl oyster mushrooms and the

commercially available antimycotic drug to a difference of merely 50% - which affirms

that it is significant from the commercially available drug. Furthermore, the study about

antimycotic property of pearl oyster mushrooms needs to continue to avoid further

damage such as lysis on red blood cells.

Keywords: antimycotic, pearl oyster mushrooms, swine, lysis, peptides, Candida spp.,

ringworms, agriculture

vi
 Table of Contents

TITLE PAGE ii

ACKNOWLEDGENMENT iii

DEDICATION v

ABSTRACT vi

TABLE OF CONTENTS vii

LIST OF TABLES ix

LIST OF GRAPHS x

LIST OF FIGURES xi

LIST OF PLATES xii

INTRODUCTION 1

Background of the Study 1

Statement of the Problem 4

Conceptual Framework 5

Hypothesis 6

Significance of the Study 6

Scope and Delimitations of the Study 7

Definition of Terms 8

vii
 Review of Related Literature and Studies 10

METHODOLOGY 21

Research Design 21

Research Instrument Used 21

Sources of Data and Data Collection 21

Data Analysis 22

Figures for Methodology 25

RESULTS AND DISCUSSION 27

Summary Table of Different Mixing and Proportions 27

Summary Table of Zone of Inhibition 28

Summary Table of Area of Healing 30

Summary Table of Amount of Time of Healing 32

Summary Table of Appearance of Swine 34

Recommendation 35

REFERENCES 36

CURRICULUM VITAE xiii

viii
 List of Tables

Table No. Title Page

4.1 Different Mixing and Proportions 27

4.2 Zone of Inhibition 28

4.3 Area of Healing 30

4.4 Amount of Time the Fungus is Totally Killed 32

4.5 Physical Appearance of the Swine after Treatment 34

ix
 List of Graphs

Graph No. Title Page

4.1 Relationship of Zones of Inhibition 29

4.2 Relationship of Area of Healing 31

4.3 Relationship of Time of Healing 33

x
 List of Figures

Figure No. Title Page

1.1 Flow Chart of the Scientific Process 5

2.1 Experimental Flow Chart 23

2.2 Method of Preparing the Mushrooms for

Experimentation 24

2.3 Default View of where the plant variable (Pearl

Oyster Mushrooms Pleurotus ostreatus)

came from. 25

2.4 Satellite View of where the plant variable (Pearl

Oyster Mushrooms Pleurotus ostreatus) came from. 26

xi
 List of Plates

Plate No. Title Page

4.1 Collecting of Pearl Oyster

Mushrooms Pleurotus ostreatus 39

4.2 Chopping of Pearl Oyster

Mushrooms Pleurotus ostreatus 39

xii
 Introduction

Background of the Study

Pearl Oyster Mushrooms Pleurotus ostreatus are mushrooms commonly seen in

the Philippines. It is an edible mushroom that grows mainly on trees. Its binomial name

came from the Latin words pleurotus (sideways) and ostreatus (oyster) which means it

grows sideways along with the stem with respect to the cap, and its cap looks like the

bivalve oyster, respectively. It is a saprotroph that is a primary decomposer of trees and

is not a parasite. It is also one of the few known carnivorous mushrooms in which its

mycelia can kill and digest nematodes, which is believed to be a way for them to obtain

nitrogen. In addition to that, pearl oyster mushrooms are high in protein, fiber, iron,

potassium, phosphorus, selenium, calcium, folic acid, and vitamins B 1, B3, B5, B12, C,

and D. Plus, pearl oyster mushrooms are low in calories, fat free, cholesterol-free,

gluten-free, and is very low in sodium. Omphalotus nidiformis, Omphalotus olivascens,

and Clitocybe dealbata are lookalikes of Pleurotus ostreatus, yet they are toxic and are

not edible.

Pork has been used for centuries as food. It supplies energy that is needed for

humans. Pig's sicknesses from pathogens are solved by humans by giving them

vitamins to increase their immune system. Yet, outbreaks are still widespread. When

there is an outbreak, the price of pork will increase due to lack of resources. The result

will not just affect the producers, but also the consumers.

Living in a tropical country, pork won't be removed from our diet. This research

was parallelized from the discussion of Mrs. Juliet A. Castronuevo (a Science teacher of

1
San Fabian National High School). She said, "Don’t you know that venom is also an

antidote for venom?”. Turns out, Snake Antivenom Immunoglobulin (simply antivenom)

are these “antidote for venom.” Antivenoms are the only specific treatment for

envenoming by snakebites. Antivenoms can prevent or reverse most of the snakebite

envenomings effects, and play a crucial role in minimizing mortality and morbidity. The

procedures for having antivenom are as follows: 1) A technician extracts and later

purifies venom from the species for which scientists want to make an antivenom; 2) A

ranch hand injects a small, harmless dose of venom into a horse. The toxins in the

venom trigger the horse’s immune system to produce antibodies that neutralize those

particular toxins; 3) Over the next year, the horse receives several booster shots with

increasing amounts of venom; 4) A ranch hand draws blood from the horse. A machine

extracts the plasma, the part of the blood that contains the antibodies. The rest of the

blood is returned to the horse; 5) the plasma is sent to a lab, where chemists purify it

and package it as a liquid or freeze-dried powder. It is then shipped in vials to hospital

pharmacies; 6) when a patient comes in with a bite or sting, doctors use an IV line to

inject the antivenom into the patient’s veins. The antibodies circulate through the body

and neutralize the toxin molecules. It is our first time to hear that and with that, we came

up with "What will happen if a fungus be an antifungal medicine?”.

The main objective of this research is to know if it is possible for Pearl Oyster

Mushrooms Pleurotus ostreatus to be an Antimycotic Drug on Pigs focusing on toxins

like Aflatoxins (by Aspergillus sp) , Ochratoxin and Citrinin (by Penicillium sp).

2
 Over the past years, mushrooms are not commercially grown for food. On the

other hand, pigs are commercially grown for pork. Thus, a decrease on the number of

pork due to unwanted pathogens will lead to increase of price. Whilst without knowing

what to do to the abundant mushrooms, will, might lead to starvation (because there are

edible and non-edible mushrooms and it’s hard to differentiate them). Further so,

economically, when supply of pork decreases, the price will increase; thus, results to a

decrease in demand.

In addition to these, the researchers ought to treat fungal infections to swine by

using natural substances which can be found in plant extracts – and not in chemically

manufactured products. Furthermore, the researchers ought to lessen human diseases

that (studies show) are caused by chemicals in chemically manufactured products and

to eliminate fear of people regarding using of chemically manufactured products

respectively.

This research aims see the result on making a fungus kill another fungus from

pigs so that outbreaks on pork will decrease and the price will be constant. It also aims

to see the other uses of the antimycotic property of pearl oyster mushrooms Pleurotus

ostreatus tested on swine and the further benefits that everyone can get from pearl

oyster mushrooms Pleurotus ostreatus.

3
Statement of the Problem

This study aims to determine the antimycotic property of pearl oyster mushrooms

Pleurotus ostreatus tested on swine. Specifically, the questions that the researchers aim

to answer are the following:

1. What are the chemical components of pearl oyster mushrooms Pleurotus

ostreatus that can be an antimycotic agent to fight fungal infections?

2. What is the effectiveness of the treatment methods indicated by:

a. Zone of Inhibition?

b. Time of healing?

c. Surface area of the fungus before and after treatment?

d. The characteristic of the pig?

3. What treatment method must be used for the infected swine?

4. What is the significant difference of antimycotic property of pearl oyster

mushrooms Pleurotus ostreatus tested on swine from the commercial one?

4
Conceptual Framework

INPUT PROCESS OUTPUT

 Collecting of Pearl
Oyster
Mushrooms
Pleurotus
ostreatus Antimycotic
 Analysis of Data
Pearl Oyster gathered from
Property of
Mushrooms Pearl Oyster Pearl Oyster
Mushrooms
Mushrooms
Pleurotus Pleurotus
ostreatus. Pleurotus
ostreatus  Testing Pearl
Oyster ostreatus
Mushrooms Tested on
Pleurotus
ostreatus on pigs Swine
with fungal
infection.
 Recording of Data.

Figure
1.FLOW

5
Assumptions and Hypothesis

This study is conducted based on the following assumptions:

1. Pearl Oyster Mushrooms Pleurotus ostreatus can be used as Antimycotic

Medication to swine.

2. Pearl Oyster Mushrooms Pleurotus ostreatus has no side effects.

3. Tested pigs in Pearl Oyster Mushrooms Pleurotus ostreatus as an Antimycotic

Medication is safe to eat.

Significance of the Study

In general, the purpose of this study is to resolve pig's illnesses caused by

fungus by using fungus too. It focuses on medicating something by the thing that

caused it. Specifically, respondents will come from piggeries and pig growers in the

town of San Fabian, Pangasinan. In addition, it will be tested on pigs that are infected

with a certain fungus (sampled Candida spp. and diverse ringworms respectively).

6
Scope and Delimitation of the Study

Because most pigs nowadays are immune to diseases, this study focuses on

swine fungal infection. Thus, this might lead for the researchers to manage the disease

and will infect the pigs mechanically and not naturally. Limits will be ensured so that

there will be no waste in livestock and economic value of the pig/hog growers of San

Fabian Pangasinan and, generally, in the Philippines. Furthermore, the researchers

sought the assistance of LDP Farms Food Corporation – Sir ____ and its stakeholders

– for supplying the researchers’ needs and technical assistance throughout the study.

7
Definition of Terms

 Antifungal – used to prevent fungal growth; active against fungi.

 Antimycotic – any agent that destroys or prevents the growth of fungi;

synonymous to antifungal.

 Antivenom – Antivenin; medication made from antibodies that is used to treat

certain venomous bites and stings.

 Candida – a yeastlike parasitic fungus that can sometimes cause thrush.

 Ex situ – means “off site.” The researchers used ex situ experimentation to test

on the off site location.

 Folic Acid – a vitamin of the B complex, found especially in leafy green

vegetables, liver, and kidney.

 Fungi – plural form of fungus; any group of spore-producing organisms feeding

on organic matter, including molds, yeast, mushrooms, and toadstool.

 Inert – lacking the ability or strength to move.

 In vivo – (of a process) performed or taking place in a living organism.

 Lysis – the disintegration of a cell by rupture of the cell wall or membrane.

 Mycelia – plural form of mycelium; the vegetative part of a fungus, consisting of a

network of fine white filaments (hyphae).

 Nematodes – roundworms; are a group of worms that make up the phylum

Nematoda.

 Organic Acids – organic compounds with acidic properties.

 Pearl Oyster Mushrooms – are the most common type of oyster mushroom.

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 Peptides – are short strings of amino acids, typically comprising 2-50 amino

acids (which are building blocks of proteins).

 Phytochemical Analysis – refers to the extraction, screening and identification of

the medicinally active substances found in plants.

 Phytopathogenic – an organism parasitic on a plant host

 Proteins – macromolecules formed by amino acids.

 Quinolones – are chemotherapeutic bacterial drugs, eradicating bacteria by

interfering with DNA replication.

 Ringworm – common fungal skin infection.

 Saprotroph – an organism that feeds on or derives nourishment from decaying

organic matter.

 Sesquiterpenes – are a class of terpenes that consist of three isoprene units and

often having the molecular formula C15H24.

 Steroids – a biologically active organic compound with four rings arranged in a

specific molecular configuration.

 Terpenes – are a large and diverse class of organic compounds, produced by

variety of plants, particularly conifers, and by some insects.

 Thrush – infection of the mouth and throat by a yeastlike fungus, causing whitish

patches; a chronic condition affecting the frog of a horse’s foot causing the

accumulation of a dark, foul-smelling substance.

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Antifungal Activity of mushroom (basidiomycetes) extracts and isolated

compounds

Abstract

The present review reports the antifungal activity of mushroom extracts and

isolated compounds including high (e.g. peptides and proteins) and low (e.g.

sesquiterpenes and other terpenes, steroids, organic acids, acylcyclopentenediones

and quinolones) molecular weight compounds. Most of the studies available on

literature focused on screening of antifungal activity of mushroom extract, rather than of

isolated compounds. Data indicate that mushroom extracts are mainly tested against

different Candida species, while mushroom compounds re mostly tested upon other

fungi. Therefore, the potential of these compounds might be more useful in food

industry than in clinics. Oudemansiella canarri and Agaricus bisporus methanolic

extracts proved to be most active mushroom extracts against Candida spp. Grifolin,

isolated from Albatrellus dispansus, seemed to be the most active compound against

phytopathogenic fungi. Further studies should be performed in order to better

understand the mechanism of action of this and other antifungal compounds as well as

safety issues.

10
Destruction of Nematodes by Pearl Oyster Mushrooms (Pleurotus ostreatus):

Abstract

On water agar, Pleurotus ostreatus produces tiny droplets of toxin from minute

spathulate secretory cells. Nematodes touching such droplets show a sudden and

dramatic response. The head region shrinks considerably; this is accompanied by

displacement of the oesophagus and (or) disruption of the tissues surrounding the

oesophagus. In as little as 30 s, and usually within several minutes, the nematode

becomes more or less immobilized but is not killed. Stimulated by leakage products

from the immobilized host, directional hyphae converge on the body orifices of the

nematode. These hyphae penetrate one or more body orifices and colonize and digest

the host. Pleurotus strigosus, P. subareolatus, and P. cornucopiae behave in a similar

way. On potato dextrose agar, P. cystidiosus produces numerous secretory cells, but

the toxin is much less potent. On water agar directional hyphae of Pleurotus ostreatus

can also be attracted to minute bacterial colonies, which are invaded and apparently

digested.

11
Potential of Oyster Mushrooms for the Biocontrol of Sugar Beet Nematode

(Heterodera Schachtii):

Abstract

The potential of oyster mushrooms to attack and kill sugar beet cyst nematode

(Heterodera schachtii) was studied. The ability of Pleurotus ostreatus, P. sajor-caju, P.

florida, P. flabellatus, P. ostreatus (sporeless), P. eryngii and Hypsizygus ulmarius to

prey on the cyst nematode was confirmed. Nematodes were inoculated on water agar

plates on which a single sparse fungal colony of one of the above mentioned fungi was

grown. Nematodes were quickly immobilized after inoculation the plates with each

Pleurotus species, with mortality of about 96%, 72%, 55%, 52%, 20%, 23% and 62%,

respectively. Nematicidal activities of culture filtrates of Pleurotus species and H.

ulmarius against H. schachtii were also studied in vitro. Extracts of P. ostreatus could

paralyse 90% of nematodes whereas P. eryngii could paralyse only 50% of nematodes.

The efficacy of old mushroom compost from cultures of P. ostreatus and P. sajor-caju in

controlling cysts on sugar beet (Beta vulgaris ICI) were studied under greenhouse

conditions. The results showed that 100 and 200 grams of mushroom compost per 3

kilogram/pot could significantly control cysts, reducing more than 85% cysts in soil.

Presence of cysts on the roots and larvae inside roots were compared with the control

treatment.

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Antifungal activity of cultivated oyster mushrooms on various agro-wastes:

Abstract

This study evaluated the antifungal activity of four fruiting bodies of oyster

mushroom harvested from three agro-substrates in vitro. At three concentrations (2, 4

and 8 mg/disc), extracts discs of Pleurotus ostreatus (grey), P. ostreatus var. florida, P.

cornucopiae var. citrinopileatus and P. salmoneostramineus were tested against three

fungal pathogens: Trichoderma harzianum (after 2 days), Verticillium sp.

and Pythium sp. (after 5 days) via the Disc Diffusion Method. The highest overall activity

was by the extract disc Y2 (P. cornucopiae grown on M2 substrate; 70% wheat straw,

20% hardwood sawdust and 10% date palm fibers) and the lowest by Y1 (P.

cornucopiae grown on wheat straw). The best inhibition zone was 16 mm toward T.

harzianum by extract disc W2 (2 mg/disc) (P. ostreatus var. florida grown on M2

substrate), compared with 23 mm with Nystatin disc (100 U), followed 7 and 5 mm by

P3 (P. salmoneostramineus grown on M3 substrate; 50% wheat straw, 30% hardwood

sawdust and 20% date palm fibers) extract disc (8 mg/disc) against Pythium sp., and (4

mg/disc) against Verticillium sp., respectively.

Keywords Pleurotus spp.; agro-wastes; contaminated fungi; antifungal activity;

bioactivity

13
Antioxidant properties in the oyster mushrooms (Pleurotus spp.) and split gill

mushroom (Schizophyllum commune) ethanolic extracts:

Abstract

Ethanolic extracts of oyster mushrooms (Pleurotus pulmonarius, P. ostreatus, P.

djamor var. djamor and P. djamor var. roseus) and the split gill mushroom

(Schizophyllum commune) were successfully investigated for their antioxidant

properties. The study was carried out to determine the capability of the extracts to

scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, to determine its reducing

ability towards ferricyanide complex (FRAP assay) and to determine the concentration

of total phenolic content (TPC) and total flavonoids content (TFC) in the macrofungi

ethanolic extracts. Inhibition concentration at 50% (IC50) for each extract to scavenge

DPPH radicals was detected from 2.75 mg/ml to 12 mg/ml where S. commune from the

fourth flush showed the lowest IC50 value among all tested mushrooms. The greatest

ability to reduce ferricyanide complex to ferrous form was observed in P. djamor var.

djamor at concentration 10 mg/ml in both first (1.23 ± 0.02) and second flushes (1.23 ±

0.00). Meanwhile, the highest TPC was determined in P. djamor var. djamor extract

(51.94 ± 0.04 mg TAE/g dry weight of extract) whereas S. commune gave highest

reading of total flavonoid content in TFC assay (29.80 ± 0.27 mg QE/g dry weight of

extract). All the mushroom samples showed appreciable antioxidant properties which

14
therefore, can be promoted as natural antioxidant preference in food and

pharmaceutical industries.

12
Key words – 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay – ferric reducing antioxidant

power (FRAP) assay – edible macrofungi – total flavonoids content (TFC) – total

phenolics content (TPC)

Pharmacological study of oyster mushroom (Pleurotus ostreatus) extract on

isolated guinea pig trachea smooth muscle:

Abstract

Mushroom farm workers suffer from respiratory symptoms during the farming of

mushrooms. The objective of this study was to analyse the effects of oyster mushroom

(Pleurotus ostreatus) extract (OME) on isolated guinea pig tracheal smooth muscle in

vitro. Isolated guinea pig tracheal tissues from 27 nonsensitized guinea pigs were

studied. The OME was obtained from indoor mushroom growing fields and prepared as

a 1:10 w/v aqueous solution. Dose-related contractions of nonsensitized guinea pig

trachea were demonstrated using these extracts. The OME contained significant

quantities of bacterial components (eg., endotoxin: 43,072.92 EU/mg). Parallel,

pharmacological studies were performed by pre-treating the tissues with mediator-

modifying agents including atropine, indomethacin, pyrilamine, BPB, acivicin, NDGA,

captopril, TMB8 and capsaicin. Atropine consistently and strikingly reduced the

contractile effects of this extract. These observations suggest an interaction of the OME

with parasympathetic nerves or more directly with muscarinic receptors. Pretreatment

15
with TMB8 (inhibitor of intracellular calcium mobilization) also significantly blocked the

constrictor effect of OME, indicating a role of calcium mobilization in the constricting

effect of OME. Inhibition of contraction by blocking of other mediators was less effective

and varied depending on the drug. We conclude that OME causes a dose-related

airway smooth muscle constriction by nonimmunological mechanisms involving a

variety of airway mediators and possibly cholinergic receptors. This effect is not

dependent on pre-sensitization of the guinea pigs.

The Effects of Temperature and Nutritional Conditions on Mycelium Growth of

Two Oyster Mushrooms (Pleurotus ostreatus and Pleurotus cystidiosus):

Abstract

The influences of temperature and nutritional conditions on the mycelium growth

of oyster mushroom Pleurotus ostreatus (PO) and Pleurotus cystidiosus (PC) were

investigated in laboratory experiment during the summer season of 2014. The results of

the experiment indicated that potato dextrose agar (PDA) and yam dextrose agar (YDA)

were the most suitable media for the mycelium growth of oyster mushroom PO while

four media (PDA, YDA, sweet potato dextrose agar, and malt extract agar medium)

were not significantly different in supporting mycelium growth of oyster mushroom PC.

The optimal temperature for mycelium growth of both oyster mushroom species was

obtained at 28℃. Mycelium growth of oyster mushroom PO was improved by carbon

16
sources such as glucose, molasses, and at 1~5% sucrose concentration, mycelium

colony diameter of mushroom PO was achieved the highest value. Whereas glucose,

dextrose, and sucrose as carbon sources gave the good mycelium growth of oyster

mushroom PC, and at 1~3% sucrose concentration, mycelium colony diameter of PC

was achieved the maximum value. Ammonium chloride concentrations at 0.03~0.09%

and 0.03~0.05% also gave the greatest values in mycelium colony diameter of

mushroom PO and PC. Brown rice was found to be the most favourable for mycelium

growth of two oyster mushroom species. In addition, sugarcane residue, acasia sawdust

and corn cob were selected as favourable lignocellulosic substrate sources for

mycelium growth of both oyster mushrooms.

Keywords: Mycelium growth, Nutritional condition, Oyster mushroom, Temperature

Disease control by chemical and biological fungicides in cultivated mushrooms:

button mushroom, oyster mushroom and shiitake:

Abstract

The most commonly cultivated basidiomycetes worldwide and in Serbia are

button mushroom (Agaricus bisporus), oyster mushroom (Pleurotus sp.) and shiitake

(Lentinus edodes). Production of their fruiting bodies is severely afflicted by fungal,

bacterial, and viral pathogens that are able to cause diseases which affect yield and

quality. Major A. bisporus fungal pathogens include Mycogoneperniciosa, Lecanicillium

17
fungicola, and Cladobotryum spp., the causal agents of dry bubble, wet bubble, and

cobweb disease, respectively. Various Trichoderma species, the causal agents of green

mould, also affect all three kinds of edible mushrooms. Over the past two decades,

green mould caused by T. aggressivum has been the most serious disease of button

mushroom. Oyster mushroom is susceptible to T. pleurotum and shiitake to T.

harzianum. The bacterial brawn blotch disease, caused by Pseudomonas tolaasii, is

distributed globally. Disease control on mushroom farms worldwide is commonly based

on the use of fungicides. However, evolution of pathogen resistance to fungicides after

frequent application, and host sensitivity to fungicides are serious problems. Only a few

fungicides are officially recommended in mushroom production: chlorothalonil and

thiabendazol in North America and prochloraz in the EU and some other countries.

Even though decreased sensitivity levels of L. fungicola and Cladobotryum mycophilum

to prochloraz have been detected, disease control is still mainly provided by that

chemical fungicide. Considering such resistance evolution, harmful impact to the

environment and human health, special attention should be focused on biofungicides,

both microbiological products based on Bacillus species and various natural substances

of biological origin, together with good programs of hygiene. Introduction of

biofungicides has created new possibilities for crop protection with reduced application

of chemicals.

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An examination of antibacterial and antifungal properties of constituents of

Shiitake (Lentinula edodes) and oyster (Pleurotus ostreatus) mushrooms:

Abstract

BACKGROUND:

Antibiotic agents have been in widespread and largely effective therapeutic use

since their discovery in the 20th century. However, the emergence of multi-drug

resistant pathogens now presents an increasing global challenge to both human

and veterinary medicine. It is now widely acknowledged that there is a need to

develop novel antimicrobial agents to minimize the threat of further antimicrobial

resistance. With this in mind, a study was undertaken to examine the

antimicrobial properties of aqueous extracts of 'exotic' Shiitake and Oyster

mushrooms on a range of environmental and clinically important microorganisms.

METHOD:

Several batches of Shiitake and oyster mushrooms were purchased fresh from a

local supermarket and underwent aqueous extraction of potential antimicrobial

components. After reconstitution, aqueous extracts were tested qualitatively

against a panel of 29 bacterial and 10 fungal pathogens, for the demonstration of

microbial inhibition.

19
RESULTS:

Our data quantitatively showed that Shiitake mushroom extract had extensive

antimicrobial activity against 85% of the organisms it was tested on, including

50% of the yeast and mould species in the trial. This compared favourably with

the results from both the Positive control (Ciprofloxacin) and Oyster mushroom,

in terms of the number of species inhibited by the activity of the metabolite(s)

inherent to the Shiitake mushroom.

CONCLUSIONS:

This small scale study shows the potential antimicrobial effects of Shitake

extracts, however further work to isolate and identify the active compound(s) now

requires to be undertaken. Once these have been identified, suitable

pharmaceutical delivery systems should be explored to allow concentrated

extracts to be prepared and delivered optimally, rather than crude ingestion of

raw material, which could promote further bacterial resistance.

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 METHODOLOGY

Research Design

This study will use descriptive-analytical method as a quantitative research

design to be able to find the effectiveness of pearl oyster mushroom (Pleurotus

ostreatus) as an antimycotic drug tested on swine.

Research Instrument Used

The researchers will use phytochemical analysis to test the pearl oyster

mushroom (Pleurotus ostreatus) in finding the isolated compounds including high (e.g.

peptides and proteins) and low (e.g. sesquiterpenes and other terpenes, steroids,

organic acids, aclycyclopentenediones and quinolones) molecular compounds as its

components which has the ability to eradicate fungus/fungi. In addition to that, the

researchers will feed the swine through the process of in vivo experimentation in an ex

situ laboratory (with the assistance of Sir ____ and the stakeholders of LDP Farms

Food Corporation).

Sources of Data and Data Collection

A total of nine (9) pigs from hog-growers in San Fabian, will be used as test subjects

where three (3) pigs will be treated with the commercial antimycotic drug, three (3) will

be treated with the drug treatment pearl oyster mushroom (Pleurotus ostreatus), and the

remaining three (3) will not be treated. The researchers will look onto the time of

21
healing, surface area of the fungus before and after the treatment, and the

characteristics of the pig. By tabular-listing method, the researchers will gather the data

needed.

Data Analysis

In this research, the researchers will use quantitative analysis for the data. The

researchers will have the extract of the mushroom and will mix it to water and the

infected swine will drink the product. On the other hand, the researchers will slice the

mushroom and mix it with the food of the swine and let the infected swine eat the

product. For the control group of swine (3), they are going to take the commercial

treatment recommended by the hog-growers.

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 Experimental Flow Chart

COLLECTING OF
PREPARATION OF
MUSHROOMS AND
DATA NEEDED
PIGSWILL

EXTRACTING AND
CLEANING OF THE
SLICING OF
MUSHROOM
MUSHROOMS

MIXING OF WATER AND

EXTRACT OF
MUSHROOM/ PIGSWILL FEEDING THE SWINES
AND SLICED
MUSHROOM

CONCLUSION AND ANALYSIS AND DATA

RECOMMENDATION INTERPRETATION

Figure
1.EXPERI

23
 Preparation of the Mushroom

GATHERING OF
MUSHROOM WASHING OF
PLEUROTUS MUSHROOM
OSTREATUS

PREPARING THE
MIXING OF WATER
GATHERED PEARL
AND EXTRACT OF
OYSTER
MUSHROOM/
MUSHROOM
PIGSWILL AND
PLEUROTUS
SLICED
OSTREATUS FOR
MUSHROOM
EXPERIMATATION

Figure 2.METHOD of PREPARING the MUSHROOMS for EXPERIMENTATION

24
 Figures for Methodology

Map where the variable is collected

Pearl oyster mushrooms (Pleurotus ostreatus) are edible mushrooms which are

grown in parts of Pangasinan – specifically in Mapandan. These mushrooms are

saprotrophs and can be seen growing on deciduous hardwoods (trees that lose their

leaves). Furthermore, these mushrooms are commercially grown for food and are being

cultured.

Figure3. DEFAULT VIEW of where the plant variable (Pearl Oyster Mushrooms Pleurotus ostreatus) came from.

Mapandan

25
 Figure4. SATELLITE VIEW of where the plant variable (Pearl Oyster Mushrooms Pleurotus ostreatus) came from.

Mapandan

The Mapandan Public Market sells this type of mushrooms which the

researchers opt to study (about its antimycotic properties and how it can help lessen the

outbreak of mycotic infections on swine). The researchers know that Mapandan can

provide their needs on upon continuing the study.

26
 RESULTS AND DISCUSSIONS

The table indicates the mixing and proportions that the researchers used in order

to find the best mixing and proportion of pure pearl oyster mushrooms (Pleurotus

ostreatus) extract with indicated amount of water (H 20). T0 is the commercially available

antimycotic treatment to swine and T 1 to T4 as the treatments which used pearl oyster

mushrooms (Pleurotus ostreatus) extract.

Amount of Concentration Amount of Water

Table1. Different MIXING and PROPORTIONS
Treatment
in Percent (%) (100 ml)
T0 CONTROL CONTROL
T1 100% 100% pure extract

T2 75% 25% H2O + 75% pure extract

T3 50% 50% H2O + 50% pure extract

T4 25% 75% H2O + 25% pure extract

The best mixing and proportion that the researchers found is 25% concentration

with 75% water and 25% pure extract. Yet, 50% concentration with 50% water and 50%

pure extract is also useable. In addition to that, the researchers also found a potent

antifungal activity with an MIC between 20% and 35% µ/ml for Candida spp.

Nevertheless, the researchers also found that certain measures must be done in order

to prevent induce lysis of red blood cells.

27
 Though Treatment 4 turned out to be the best mixing and proportion, the

researchers sought to inform other researchers that late studies shouldn’t be

disregarded. Thus, induced lysis on red blood cells must be considered.

The table shows the zone of inhibition of the fungus/fungi (specifically Candida

species) on swine. The columns show the results of treatments T0 to T4 and to show the

differences of different methods the researchers used.

Table2. Zone of INHIBITION

Zone of Inhibition (mm)
Treatment Trial 1 Trial 2
R1 R2 R3 R1 R2 R3
T0 CONTRO CONTRO CONTRO
CONTROL CONTROL CONTROL
L L L
T1 170 150 140 160 150 170
T2 160 190 170 200 180 150
T3 210 180 200 190 220 190
T4 220 210 230 240 200 220

In T1 the researchers used 100% pure pearl oyster mushrooms (Pleurotus

ostreatus) extract which resulted to a mean of 156.67. In T 2 the researchers used 75%

pure pearl oyster mushrooms (Pleurotus ostreatus) extract which resulted to a mean of

175. In T3 the researchers used 50% pure pearl oyster mushrooms (Pleurotus ostreatus)

extract which resulted to a mean of 198.33. In T 4 the researchers used pure pearl oyster

mushrooms (Pleurotus ostreatus) extract which resulted to a mean of 220.

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 Graph1. RELATIONSHIP of Zones of Inhibition

The graph shows the relationship of the results from trial 1 to trial 2 and the

average mean of the two trials related to the 4 treatments – excluding the commercially

available antimycotic treatment.

29
 The table shows the area of healing (indicated by millimeters). Commercially

available anti-mycotic drugs are used on Swine A, B and C; pearl oyster mushrooms

(Pleurotus ostreatus) on Swine D, E and F; and Swine G, H and I were not treated.

Negative numbers are used by the researchers to show that the area healed was made

worse. Hence, was made worse. Area of healing is all measured 1 day after the first

treatment.

Sample
Table3. Swine
Area of HEALING Area of Healing Average Area
(mm) (mm)
T1 T2 T3 Tave
Swine A 105 70 140 105
Swine B 105 175 245 175
Swine C 140 175 105 140
Swine D 175 140 210 175
Swine E 245 210 175 210
Swine F 210 175 175 186.67
Swine G -35 -140 -140 -105
Swine H -35 -175 -105 -105
Swine I -35 -70 -140 -81.67

In Swine A, the T1, T2 and T3 have the average of 105. In Swine B the T1, T2 and T3

have the average of 175. In Swine C the T 1, T2 and T3 have the average of 140. In Swine

D, the T1, T2 and T3 have the average of 175. In Swine E the T 1, T2 and T3 have the

average of 210. In Swine F the T1, T2 and T3 have the average of 186.67. In Swine G the

T1, T2 and T3 have the average of -105. In Swine H the T1, T2 and T3 have the average of -

105. In Swine I the T1, T2 and T3 have the average of -81.67.

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 Graph2. RELATIONSHIP of Area of Healing

The graph shows the relationship of the results from trial 1 to trial 3 and the

average mean of the three trials related to the 9 swine.

31
 The table shows the amount of time the fungus (in the swine) is totally killed –

indicated by day/s. Commercially available mycotic drugs are tested in Swine A, B and

C. Pearl oyster mushroom (Pleurotus ostreatus) are tested in Swine D, E and F. Swine

G, H and I, however, were not treated. Negative numbers are used by the researchers

to show that the time of healing was made worse. Time of healing is measured 1 day

after the first treatment.

Table4. Amount of TIME the Fungus is totally KILLED
Sample Swine Amount of Time the Fungus is Totally Killed (days)

R1 R2 R3 Rave

Swine A 4 3 5 4

Swine B 4 6 8 6

Swine C 5 6 4 5

Swine D 6 5 7 6

Swine E 8 7 6 7

Swine F 7 6 6 6.33

Swine G 0 -3 -3 -2

Swine H 0 -4 -2 -2

Swine I 0 -1 -3 -1.33

The commercially available anti-mycotic drug averaged 5 in Swine A, B and C.

Pearl oyster mushrooms (Pleurotus ostreatus), however, averaged 6.44 in Swine D, E

and F. Additionally, the Swine G, H and I averaged -1.78.

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 Graph3. RELATIONSHIP of Time of Healing

The graph shows the relationship of the results from replication 1 to replication 3

and the average mean of the three replications related to the 9 swine.

33
 The table shows the live weight (indicated by kg), color (indicated by vibrancy)

and behavior (indicated by active or inert) of the Samples that the researchers used –

from Swine A to Swine I – using the commercially available anti-mycotic drug in Swine

A to C, pearl oyster mushrooms in Swine D to F, and no treatment to Swine G to I. The

results explain the physical appearance of the swine after the treatment .

Table5. PHYSICAL APPEARANCE of the Swine after Treatment

Sample Live Weight (kg) Color (vibrancy) Behavior (active

or inert)

Swine A 76 Glowing Active

Swine B 79 Glowing Active

Swine C 77 Glowing Active

Swine D 78 Bright Active

Swine E 74 Bright Active

Swine F 76 Bright Active

Swine G 69 Pale Inert

Swine H 71 Pale Inert

Swine I 70 Pale Inert

Swine A, B and C are active, glowing and didn’t lose weight which means the

quality is above. Swine D, E and F are active, bright and partially lose weight which

means the quality is just right. However, in Swine G, H and I are inert, pale and lost

34
weight which means they didn’t met the requirement to fully irradiate the fungi/fungus

infection.

Further studies about Pearl Oyster Mushrooms (Plerotous ostreatus) are ought to

be furnished by the researchers. Though studies already show its antimycotic abilities,

to prevent harm, deeper studies must be done to not cause harm.

35
 References

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 Potočnik, I., Stepanović, M., Rekanović, E., Todorović, B., & Milijašević-Marčić, S.

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 Matejuk, A., Leng, Q., Begum, M.D., Woodle, M.C., Scaria, P., Chou, S-T., &

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L

T
38
 E
Collecting of Pearl Oyster Mushrooms Pleurotus ostreatus

Chopping of Pearl Oyster Mushrooms Pleurotus ostreatus

39