Evaluate the effect of honey and choerospondias axillaris mixture on necrotic bacteria (khóa luận tốt nghiệp)

VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE
FACULTY OF BIOTECHNOLOGY
-------  -------

THESIS
EVALUATE THE EFFECT OF HONEY AND
CHOEROSPONDIAS AXILLARIS MIXTURE ON
NECROTIC BACTERIA

Hanoi, 2021


VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE
FACULTY OF BIOTECHNOLOGY
-------  -------

THESIS
TITLE
EVALUATE THE EFFECT OF HONEY AND
CHOEROSPONDIAS AXILLARIS MIXTURE ON
NECROTIC BACTERIA

Student

: Bui Duc Hiep

Faculty

: Biotechnology

Supervisors

: Nguyen Thi Thuy Hanh, PhD.

Hanoi, February 2021


COMMITMENT
I hereby declare that:
This is my study, which was conducted under the guidance of supervisors;
All of information provided in this study is true and accurate
All published data and information have been duly cited.
Hanoi, February 2021
Student

Bui Duc Hiep

i


ACKNOWLEDGEMENTS
First, I am grateful to express my gratitude to the Thai Nguyen University of
Medicne and Pharmacy for granting me support to pursue my thesis as well as the
Department of Biotechnology, Vietnam National University of Agriculture, who
provided me opportunities to pursue a higher education, anh to prepare myself to
better.
I also would like to thank my supervisor, Dr Nguyen Thi Thuy Hanh who has
given me her encouragement, and help during my writing and revising process.
I owe a debt of gratitude to many people whose help has been important to my
success in completing this dissertation. Firstly, I am deeply grateful to my major Bs.
Nguyen Thi Hien and Mas. Nguyen Van Dung for their critical comments and

useful suggestion. Special for their help in collectiong data of my experiments.
It would be difficult to mention everyone who has contributed to the research
reported in this dissertation. I appreciate you all. It is also give my thankfulness to
my family giving me various used advises during the process of learning and
studying
Thank you very much!
Hanoi,February 2021
Student

Bui Duc Hiep

ii


TABLE OF CONTENTS

COMMITMENT........................................................................................................................... i
ACKNOWLEDGEMENTS ....................................................................................................... ii
TABLE OF CONTENTS...........................................................................................................iii
LIST OF TABLES ....................................................................................................................... v
LIST OF FIGURES ...................................................................................................................vii
ABSTRACT ........................................................................................................... viii
I. INTRODUCTION ................................................................................................................. 1
1.1. Problem: ................................................................................................................................. 1
1.2. Evaluate the effect of Honey and Choerospondias axillaris mixture on necrotic bacteria........1
1.3. Purpose and Requirement..................................................................................................... 2
1.3.1 Purpose ................................................................................................................................. 2
1.3.2 Requirement .......................................................................................................2
II. OVERVIEW .......................................................................................................................... 3
2.1 Introduction of Choerospondias axillaris ............................................................................ 3

2.1.1 General introduction of Choerospondias axillaris .......................................................... 3
2.1.1.1. Distribution ...................................................................................................................... 3
2.1.1.2. Chemical composition .................................................................................................... 4
2.1.2 Botanical characteristic of Choerospondias axillaris ...................................................... 6
2.2. Honey ..................................................................................................................................... 8
2.3 Necrosis of human skin ....................................................................................................... 10
2.3.1 General information .......................................................................................................... 10
2.2.2 The main types of bacteria cause necrosis of human skin ............................................ 11
2.2.2.1. Streptococcus................................................................................................................. 11
2.2.2.2. Pseudomonas aeruginosa ............................................................................................. 13
2.3 The application of Choerospondias axillaris on folk drug .............................................. 15
III: MATERIALS AND METHODS .................................................................................. 17
3.1 Materials ................................................................................................................................17
3.2 Time and place...................................................................................................................... 17

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3.3 Methods .................................................................................................................................17
3.3.1. Extraction of material from Choerospondias axillaris.................................................. 17
3.3.2.Implementation of experiment In Vitro .......................................................................... 18
3.3.3. Implementation of experiment on white mice .............................................................. 19
3.4 Experiments to assess the ability to kill bacteria Streptococcus and Pseudomonas
aeruginosa cause skin necrosis of honey products and Choerospondias axillaris on white
mice ..............................................................................................................................................20
IV. RESULTS AND DISCUSSION ..................................................................................... 22
4.1 Extraction results for Choerospondias axillaris ................................................................ 22
4.2 Test in vitro .........................................................................................................22
4.2.1 Research results on properties of Streptococcus of Honey and Choerospondias
axillaris on a sterile ring ...........................................................................................22

4.2.2 Research results on antibacterial properties of Pseudomonas aeruginosa of
Honey product + Choerospondias axillaris extract on sterile ring ...........................24
4.2.3. Research results of the experiment determine the dose of bacteria that can
cause necrosis on an individual mouse .....................................................................27
4.2.3.1. Streptococcus ...............................................................................................27
4.3.2. Pseudomonas aeruginosa ............................................................................................... 29
4.4. Research results of the experiment evaluating the ability to kill Streptococcus and
Pseudomonas aeruginosa cause skin necrosis of honey products - Choerospondias
axillaris in white mice ................................................................................................................ 33
V. CONCLUSION AND PROPOSAL .....................................................................35
5.1. Conclusion..........................................................................................................35
5.2 Proposal .................................................................................................................................35
REFERENCE ............................................................................................................................. 36

iv


LIST OF TABLES

Table 3.1. Mix with honey according to different ratios to get 5 formulas
for studies ............................................................................................ 18
Table 3.2 Evaluate the general situation of white mice before causing
streptococus ........................................................................................ 20
Table 3.3 Evaluate the general situation of mice before causing illness by
the Pseudomonas aeruginosa ............................................................. 20
Table 3.4 Assess the general situation of mice before causing disease .......... 21
Table 4.1 Results of bacterial inhibition loop on 3 experimental agar with
Streptococcus ...................................................................................... 23
Table 4.2. Variance and standard deviation of the formulas Resistant to
Streptococcus ...................................................................................... 23

Table 4.3. Anti-streptococcal effect of Preparations of Honey Choerospondias axillaris........................................................................... 24
Table 4.4. Results of bacterial inhibition loop on 3 experimental agar
plates with Pseudomonas aeruginosa ................................................. 25
Table 4.5. Variance and standard deviation of the formula resistant to
Pseudomonas aeruginosa ................................................................... 25
Table 3.1. Mix with honey according to different ratios to get 5 formulas for studies ................... 18
Table 3.2 Evaluate the general situation of white mice before causing streptococus ................... 20
Table 3.3 Evaluate the general situation of mice before causing illness by the Pseudomonas
aeruginosa........................................................................................................................................ 20
Table 3.4 Assess the general situation of mice before causing disease .......................................... 21
Table 4.1 Results of bacterial inhibition loop on 3 experimental agar with Streptococcus ............. 23
Table 4.2. Variance and standard deviation of the formulas Resistant to Streptococcus ............... 23
Table 4.3. Anti-streptococcal effect of Preparations of Honey - Choerospondias axillaris ...................... 24
Table 4.4. Results of bacterial inhibition loop on 3 experimental agar plates with Pseudomonas
aeruginosa........................................................................................................................................ 25

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Table 4.5. Variance and standard deviation of the formula resistant to Pseudomonas aeruginosa25

vi


LIST OF FIGURES

Figure 2.1 Leaves and Fruits ............................................................................. 6
Figure 2.2 Young Leaf ...................................................................................... 7
Figure 2.3. Honey .............................................................................................. 8
Figure 2.4. Necrosis of the hand ..................................................................... 11

Figure 2.5. Necrosis of the feet ....................................................................... 11
Figure 2.6. Butt necrosis ................................................................................. 11
Figure 2.7. Leg necrosis .................................................................................. 11
Figure 2.8. Streptococcus ................................................................................ 12
Figure 2.9. Pseudomonas aeruginosa .............................................................. 14
Figure 3.1. Medicinal herbs have been dried .................................................. 17
Figure 3.2. Measure the diameter of the punching tool and the agar was
perforated ............................................................................................ 19
Figure 4.2. Measure the bacterial inhibitory ring for the bacterial
connective system ............................................................................... 22
Figure 4.3. Direct antibacterial resistance of regime ...................................... 25

vii


ABSTRACT
Nowadays, the use of natural products to treat necrosis is of great interest to
many scientists and pharmaceutical companies. The use of products derived from
nature shows many advantages over the use of compounds and drugs that are
synthesized by chemical methods. namely environmentally friendly, safe for users,
low irritation rate, etc. However, besides that, it also has some limitations such as
the need for an abundant source of raw materials, not much. in-depth research
works on those preparations. Most natural preparations used to treat necrosis are on
the basis of folk remedies.
Based on these advantages, thesis “Evaluate the effect of Honey and
Choerospondias axillaris mixture on necrotic bacteria” was carried out to study
and evaluate the effect in the treatment of necrosis disease for two main factors,
streptococcus and Pseudomonas aeruginosa.
The research results of the project show that the product from honey and
choerospondias axillaris can treat necrosis in in vitro experiments and experiments

performed on living individuals (white mice). The medicinal use of the preparation
is outstanding and is likely to be widely used in the future.

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I.

INTRODUCTION

1.1. Problem:
In today's life, there are many dangerous diseases. Among the 15 most
dangerous diseases, many lead to necrosis of the skin. Skin necrosis are necrotic
ulcers that damage the most basic defense system, the patient's body is in pain and
cannot resist the negative effects from outside, so ulcers can cause blood infection
... threatens the patient's life. The main cause of necrosis is a decrease in the blood
supply to damaged tissues and cell death. [6]
There are many different causes of skin necrosis, which can be named to the
most basic ones, which are: Necrotic skin ulcers due to occlusion of the arteries;
Peripheral neurological complications of diabetes; Cerebral vascular accident
patient with human paralysis complications; Infected boils cause necrotic skin
ulcers.
To date, there have been a number of domestic and foreign studies on
necrosis problem and necrosis treatment methods in different parts of the human
body. For example, the authors of the Hanoi Medical University Hospital conducted
a study since 2012 on necrotizing fasciitis (necrotizing fasciitis). This is a bacterial
infection usually caused by streptococcus or staph. Research has concluded: skin
necrosis, muscle weight due to infection is a rare disease. High mortality if not
treated promptly. Combining medical treatment against infection, cleaning lesions,
and skin patching is a treatment with good results. [1]

1.2. Evaluate the effect of Honey and Choerospondias axillaris mixture on
necrotic bacteria
Regarding the herbal necrosis treatment drugs, I had access to information
about Nacurgo products containing Polyesteramide biofilm, Nano Curcumin and
Camellia Sinensis green tea extract to help disinfect the stain. skin ulcers, allowing
skin ulcers to heal 3 to 5 times faster than normal and do not lead to serious
consequences such as sepsis leading to necrosis. [11]
Traditionally, honey or products from choerospondias axillaris are used as
support for the treatment of necrosis. In fact, there is also no research and

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evaluation on the combination of honey with choerospondias axillaris in
bactericidal and necrotic treatment.
Therefore, our group conducted research project to evaluate the bactericidal
ability with honey and polymers. This research is conducted to provide more
scientific evidence on the treatment of necrosis of honey and extracts of
choerospondias axillaris, contributing to opening a new direction in research on
drug formulation and Treatment for necrotic patients from the very plants and
animals available in Vietnam.
1.3. Purpose and Requirement
1.3.1 Purpose
Provides scientific evidence of a folk remedy using choerospondias axillaris
extractum in the treatment of necrotic ulcers caused by infections.
1.3.2 Requirement
- Extraction of material from Choerospondias axillaris
- Implementation of experiment In Vitro
- Implementation of experiment on white mice

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II.

OVERVIEW

2.1 Introduction of Choerospondias axillaris
2.1.1 General introduction of Choerospondias axillaris
Scientific name: Choerospondias axillaris (Roxb.) Burtt et Hill (Spondias
axillaris Roxb.), Belongs to the Anacardiaceae.
Other names: Oval tea, wood, forest oval, sliced oval, toads, male acid apple.
Scientific classification
Kingdom: Plantae
Order: Sapindales
Family: Anacardiaceae
Genus: Choerospondias
Species: C. axillaris
2.1.1.1. Distribution
Meliaceae (Meliaceae family) is a family of plants mainly tree trunks with
about 50 varieties, 550 species distributed throughout the tropics. In the world, this
plant is distributed in South Asian countries such as India, Nepal ...; Southeast
Asian countries such as Thailand, Malaysia, Indonesia, Laos, Cambodia and some
southern provinces of China such as Guangdong, Hubei, Hunan, Guangdong,
Guangxi, Yunnan, Fujian. In Vietnam, Choerospondias axillaris is quite popular in
most provinces across the country. Provinces with many Choerospondias axillaris
include: Ha Giang, Tuyen Quang, Yen Bai, Lao Cai, Lang Son, Vinh Phuc, Ha Tay,
Ninh Binh, Quang Tri, Dak Lak, Gia Lai, Kon Tum ... and especially in Thai
Nguyen.
In the midland and mountainous areas in general and Thai Nguyen province

in particular, the oval tree grows popularly due to favorable climatic and soil
conditions: it is a tropical climate with rainfall over 1500mm / year and seasonal
distribution; Slope topography is gentle, easy to drain with soils still forested in
nature, moist, less acidic, deep and thick. [9]

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2.1.1.2. Chemical composition
Chemical studies show that the oval family contains triterpenoid tirucallan,
dramaran, oleanan, multifloran and limonoid. In addition, the rhizome bark and
rhizome contain a bitter alkaloid (formula C9H8O4) that has a worm-killing effect.
Furthermore, the fruit contains an alkaloid called azaridin and an oil (60%), the oil
has sulfur and the smell of garlic. The leaves contain an alkaloid called paraisin, a
small amount of rutin (0.5% dry matter).
In 1975, Nguyen Liem and his colleagues analyzed the mollusc bark and
found that there were 37.1% pyrogalic tannins, 5.4% flavones, 0.6% quirions, and
14% natural polymers.
In particular, in the bark and root bark of the oval tree, it still contains 70%
tannin (13.7% in stem bark; 1% in stem wood; 44.8% in root bark and 1.9% in root
wood; , 5%); Flavonoids; Quinon; Plastic gum; Fatty oil. In the leaf bark about 1314% gum gum. [3]
To serve the purposes of the topic, from the above chemical components of
the oval tree, we pay special attention to the following three compounds: tannin,
flavonoid and quinon. The reasons the above compounds play an important role in
research are:
Tannins: Tannins are natural compounds in the polyphenol group common in
plants. They are acrid and have tanning properties. This means being able to bind to
the proteins of the skin to form a structure that is stable to the decay process. The
molecular weight of tannin is about 1,000 - 5,000.
(The term "tannin" is used in the biological and food industries to refer to all

natural polyphenols that have an acrid taste, but not these substances have a true
tanning ability. This property is only available to compounds. High molecular
weight molecules from 1,000 - 5,000. The molecules with lower molecular weight
have only acrid taste without tanning properties, to distinguish it is called "food
tannin", "tea tannin" .)
In the plant, tannin is involved in metabolism, redox processes. As
polyphenols, tannins have antibacterial properties, so they play a protective role for

4


the plant. In particular, tannin solution combines with protein, forming a membrane on the
mucosa, so it is used as a tonic. Tannin also has an antibacterial effect, so it should be used
as a mouthwash when the lining of the mouth, throat is sore, or when the ulcer is long.
Tannin can be used to treat inflammatory bowel disease, diarrhea ...
Flavonoids: Flavonoids are compounds found in plants, and more than half of
them are vegetables and fruits that we often use. In plants, Flavonoids act as antioxidants,
"guard" and preserve ascorbic acid in cells, inhibiting certain pathogens harmful to plants
(bacteria, insects, ...). Also work is to regulate the growth of plants. Flavonoids are also a
common ingredient in plant-derived medicinal products.
Most flavonoids in the human body act as antioxidants. With this ability,
they help neutralize over-reactive oxygen-containing molecules and prevent those
molecules from harming cells. Especially in oriental medicine, flavonoid-containing
plants are used a lot for antioxidants such as licorice or green tea ...
Dr. Albert Szent-Gyorgi, who won the Nobel Prize, discovered a link
between flavonoids and vitamin C when he treated a patient with blood vessel
health problems. When he used only pure vitamin C ingredients (100% vitamin C,
no other compounds) he found little effect on the patient. But when he used a
combination of vitamin C and flavonoids, the results were surprising. Since then,
through many other studies, Dr. Albert confirmed the need for flavonoids for

vitamin C: this substance actively supports the action of the other in antioxidant.
And later research also shows the need for flavonoids for the activities of other
vitamins. In addition, we also know that inflammation is a natural response to the
body and needs to be carefully monitored to prevent overreaction of the immune
system and other undesirable effects. Many types of cells involved in the immune
system such as T cells, B cells, NK cells or neutrophils have had a positive change
in the presence of flavonoids. Chemical compounds with flavonoids are considered
as one of the key keys in preventing and controlling overgrowth. In many cases,
flavonoids can act directly as antibiotics by disrupting functional structures of
microorganisms such as viruses, bacteria, etc. The antiviral function of flavonoids
has been proven against HIV. or HSV-1, a virus that causes herpes sores ...

5


Quinone: Quinone or benzoquinone is

one of two isomers of

xyclohexadienedion. They have the general chemical formula C6H4O2.
Orthobenzoquinon is 1,2-dion, while parabenzoquinon is 1,4-dion.
Orthobenzoquinone is the oxidized form of catechol (1,2-dihydroxybenzene),
while parabenzoquinone is the oxidized form of hydroquinone. The potassium
iodide solution reduces the benzoquinone solution to hydroquinone and it is
oxidized again with silver nitrate solution.
2.1.2 Botanical characteristic of Choerospondias axillaris
Choerospondias axillaris is a large, 8 ~ 20m tall tree with gray-brown bark.
Large, deciduous tree in the dry season, canopy, broad, 15-25 m high, 40-60 cm in
diameter or more, straight stem, very thick bark, gray-brown or pinkish brown,
longitudinally cracked and flaking into an array (looks like a flower slice). The flesh

is 2 cm thick and has gray plastic. Young shoots are black-brown or purple-brown
with many light brown skin. Single feathered compound leaves 1 time, growing
apart, 30-40 cm long, with 7-15 leaflets. The leaflets are opposite, 5-10 cm long, 24.5 cm wide, ovoid, lanceolate or oval. . Flower heterosexual, different original. The
male flowers fake purple red, growing into a cluster of flowers at the axillary or
branch head 4-12 cm long. The female flower is solitary, grows in the armpit; sepals
5; petal 5; second 10, vote 5 boxes. Nuts, shaped like 2 to 3 cm long, shiny, yellow,
top with 5 small holes, sour taste, sweet taste when ripe. Lean fruits are ovoid or
globose, 2-3 cm long, 1-1.5 cm wide, brown yellow when ripe, with edible flesh.
Hard seeds have 5 holes at the top, usually carrying 2-4 fertile embryos.

Figure 2.1 Leaves and Fruits
(Source: Internet)

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This plant likes light, but under 3 years old is slightly shade tolerant; grows
fast, especially in the 1-5 year old period. Average growth in height is 1.5-2m / year
and growth in diameter is 1.5-2cm / year. Trees 5 years old can reach a height of 7-8
m and a diameter of 7-10 cm; 10 years old tree, the diameter can be up to 20 cm.
Then the growth rate decreased gradually. The root system is mainly cluster roots,
eating about 3-4 m wide in diameter and 50-70 cm deep. Plants have a very good
regeneration ability with seeds and shoots. The 7-year-old plant begins to produce
fruit. Flowering season is from March to May; at the same time young leaves
appear; Fruit season is from July to October of the year.
The tree produces a light yellow-pink color, has a beautiful straight pattern,
has a soft, light texture, is easy to work with and is durable, is resistant to termites,
termites, so it is often used as a house, closes common furniture ...

Figure 2.2 Young Leaf

(Source: Internet)

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2.2. Honey
Honey is a product created from worker bees by harvesting from the nectar
glands of flowers or nectar plants - Nectar is a dilute solution of sugar that is sucked
into the gallbladder by worker bees and mixed with gallbladder enzymes to produce
honey. is a very high calorie product.

Figure 2.3. Honey
(Source: Internet)
By modern techniques have identified in honey contains at least 100
substances with biological effects important to the human body. The main
ingredients present in honey mainly include:
Water content: from 18.5-22%, water content 22% higher than bile is usually
fermented and easily spoiled. [8]
Sugar content: The main component of honey is sugar, in which glucose and
fructose account for 85%, sucrose sugar accounts for 1-5%, the rest are double
sugars such as mantose, isomaltose, nigerose, turanose, maltulose, leucrose. ,
neotrehalose, gentiobiose, laminaribiose, and small concentrations of other poly
sugars. [8]
Protein and amino acids: Protein content in honey is about 0.1-3.0%, these
enzymes come from pollen and are secreted by bees during honey production, such
as (diastase, invertase, glucose oxidase , catalase). [8]

8

Enzymes: Honey is considered to be a food containing many enzymes, these
enzymes play a very important role for the body. If lacking, the body cannot digest and
absorb nutrients from food. The enzymes diastase, invertase, glucose oxidase, and acid
phosphatase are only available in a small amount from the raw materials that bees
harvest from nature, and most are secreted by bees during honey refining. [8]
The acids present in honey include: gluconic, acetic, butyric, lactic, citric,
formic. The content of formic acid in nectar ranges from 50-200 ppm, leaf nectar
from 200-500 ppm. [8]
Minerals in honey: According to the analysis of Baden - Wurtemberg (1987)
in honey, there are elements (Potassium, Sodium, Calcium, Copper, Magnesium,
Manganese, Chlorine, Sulfur, Phosphorus, Iron, Bo , Aluminum). [8]
Vitamins: The content of vitamins in honey is not much, but there are many
types, for every 1 kg of honey has: 1.5mg B2, 2mg B3, 1mg PP, 5mg B6 and 3054mg vitamin C. [8]
Colorants and aromatics: The scent of honey is caused by amino acids, plant
hormones and plant growth hormones that bees take back to the hive for processing
into honey. [8]
Measure the color of honey: The color of honey can vary widely, ranging
from lemon yellow, amber, yellow, light brown or golden brown, to dark brown,
depending on the type of tree that produces the honey. [8]
Today's medicine has proven honey's ability to heal ulcers. In the honey
composition, albumin and panthotenic acid participate in the process of forming and
constructing new cells, thus helping to quickly repair damage. TPO - Researchers at
the Cochrane Global Research Organization have concluded that honey can be used
as a remedy for burn wounds.
Dr Andrew Jull from the Center for Clinical Trial Research at the University
of Auckland, New Zealand, said: “We are still studying and testing the results
carefully, but honey definitely works. heal some burn wounds ”.

9

Honey has been known for this use for a long time, however, there are no
clear grounds to draw a definite conclusion. Current research focuses on honey's
ability to repel bacteria.
The study summary has presented data of 19 experimental treatments,
including 2,554 patients with different levels of injury. In particular, honey is used
to heal wounds faster than some types of bandages and gauze commonly used to
treat similar wounds.
2.3 Necrosis of human skin
2.3.1 General information
There are many different causes of skin necrosis, which can be named to the
most basic ones, which are: Necrotic skin ulcers due to occlusion of the arteries;
Peripheral neurological complications of diabetes; Cerebral vascular accident
patient with human paralysis complications; Infected boils cause necrotic skin
ulcers. The large statistical results show that the number of patients with necrosis
due to cerebral vascular accident leading to paralysis accounts for the highest
percentage among the four groups of basic causes above. According to research
from medical experts, if a person stays in the same position for more than 2 hours a
day, it can form necrotic ulcers. The average person unfortunately has a stroke that
can cause a nerve complication that causes paralysis, all activities depend on family
members. If the patient lies for a long time and is not changed position, necrotizing
ulcers will form in the heels, buttocks, shoulders [6] ...
Some pictures (Source: Internet) below illustrate the common necrosis cases
in patients due to different causes:

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Figure 2.4. Necrosis of the hand

Figure 2.5. Necrosis of the feet

Figure 2.6. Butt necrosis

Figure 2.7. Leg necrosis

2.2.2 The main types of bacteria cause necrosis of human skin
There are many types of bacteria that cause necrosis on different parts of the
human body. Causing necrosis of human skin, there are three main types of
bacteria: Streptococcus, Pseudomonas aeruginosa [2]
2.2.2.1. Streptococcus
Shape and color-catching properties, streptococcus bacteria are spherical,
standing in long or short chains, sometimes individually, catching the color of
Gram-positive. Streptococcus is not mobile, has no shell, does not produce spores.

11


Figure 2.8. Streptococcus
(Source: Internet)
Binding is optional aerobic bacteria, which can grow on normal culture
media but thrive on blood, serum, semolina, and gases typically with 5-10% CO2.
Suitable temperature is 370C, pH = 7.2- 7.4.
Straight environment, at 370C / 24h conditions, the growth link does not
obscure the environment, often reconnects to the bottom environment, the above
environment becomes clear. Especially in the above environment, at 370C / 24h, the
link develops into the form of F: small, clear, glossy, glossy, evenly shaded, in fog.
In the above environment agar, at 370C / 24h, the link develops into a small
system, around with dissolved blood. There are 3 types of hemolytic:
+ Tan (α): Lac bare is surrounded by a large, clear, completely dissolvable,

blood-limited part.
+ Hemolysis (β): The colony is surrounded by a dissolving, incomplete, blue
blood.
+ Hemolysis (γ): also known as hemolysis.
Biological and chemical properties
Binding without catalase enzyme, insoluble in salt and pigment density
(standard for binding analysis), on milking medium, on gelatin medium, not to
damage gelatin. The A-bonding group dissolves in sensor type with Bacitracin.
Create an integer and group the partition

12


C (Polysaccharide) antigen: is a component of stem bacteria, has a
characteristic group. On this raw linkage link divided into groups A, B, C, D ... In
which, the most important is group A linked, hemolytic type. M (Protein) antigen:
This resource is for allowable type is used in the same group. Group A has category
12, of which category 12 is the most important. T (Protein) antigens. R
(NucleoProtein) antigen: This substance keeps the septum of the bond hard and is
also toxic of bacteria.
Toxicity factor: Hemolysin (Hemolysin): is a mixture of 2 substances:
Streptolysin O and Streptolysin S. In which, most of the hemolytic link produces
Streptolysin O enzyme. It is easy to lose oxygen function. , which is of the nature of the
exotoxin adaptive facility, forms a possible inductance against Streptolysin O. In the
required acute rheumatism and inflammation testing, the quantitative work may mean the
positioning device Stability More stable is Streptolysin S, which has the ability to
dissolve blood even on the surface of the medium, but the antigenic factor is active.
Streptokinase: has a fibrinolytic effect, activates around the commercial area,
helps to spread faster.
Hyaluronidase: yeast that hydrolyzes the organization's hyaluronic acid to

help bacteria spread more easily. This men also has an antigen, a possible trigger
basis for induction. Erythrotocine: Reddening toxin in scarlet fever. Proteinase:
Protein breakdown effect. Among the species capable of causing disease, the most
important one is Str.pyogens (group A):
+ Skin infections: acne, dermatitis, wound dermatitis.
+ Inflammation, inflammation, inflammation.
+ Urinary tract, genital tract. Rheumatism, low heart rate, subacute endocarditis,
septicemia. Enteritis (usually caused by anti-inflammatory group D). [13]
2.2.2.2. Pseudomonas aeruginosa
Special biology
Appearance: Direct, round head, 1-5 µm long, 0.5-1 µm wide. Less online
when shelled, has a hair on the head, is mobile, does not produce cells, catches
Gram-negative color.

13


Figure 2.9. Pseudomonas aeruginosa
(Source: Internet)
Pseudomonas aeruginosa
Culture properties: Bacteria grow easily on normal, aerobic culture media.
Suitable temperature is 37 ° C, growing at a temperature of 5-42 ° C. On a solid
medium, there are usually two types of colonies, one large, smooth, flattened, with a
slightly convex center, and a small, rough, convex type.
Pigmentation: The characteristic properties of the green pus bacteria are
pigmentation and aroma. There are two main types of pigments, pyoxianin, which is
blue, water-soluble, and Cholorofor, which give cultures and colonies green.
Pyoverdin is a fluorescent pigment, soluble in water but insoluble in Cholorofor.
The pigments of the Pseudomonas aeruginosa are Phenazin derivatives, under the
chemical influence they can change the pigmentation brown, black, red, yellow. The

aroma produced by the bacillus is Kimethylamin. Biological and chemical
properties: Glucose fermentation. No fermentation of lactose. Oxydase (+), Citrate
simmons (+). Indole (-), H2S (-), LDC (-).
Antigen structure:
Hair antigen H: common to the whole breed, this antigen is easily destroyed
by temperature.
O-body antigens: specific for each type, O-antigens are stable with
temperature, essentially a complex of carbohydrates-lipid-proteins, similar to
endotoxins of intestinal bacteria.
Ability to cause illness to people. Pseudomonas aeruginosa are bacteria that
cause conditions that cause disease. Therefore, infection often occurs in people

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whose defense mechanisms are compromised such as long-term use of
corticosteroids or antibiotics, severe burns or intravenous drugs ... The usual site of
infection is an open wound. , is one of the causes of skin necrosis. At the site of
entry, this bacterium causes purulent inflammation (blue pus), the body has reduced
resistance, can penetrate deeper into the body and cause inflammation of the viscera
such as: purulent infections and abscesses in different parts of the human body.
Cases of endocarditis, pneumonia, meningitis, although rare but also occur or cause
systemic illness (such as sepsis, infection in newborn babies or premature births are
often very serious). Septicemia often occurs in debilitated individuals.
In recent years, Pseudomonas aeruginosa infection has become important due to
the treatment and prevention of infections with antibiotics to which the bacteria are
resistant, corticosteroids, anti-metabolic drugs and immunosuppressants. has reduced the
body's resistance, using examination tools such as catheters and other instruments to
probe that is not disinfected well. Blue pus bacillus is a notable cause of hospital
infections such as postoperative infection and severe burns. Pseudomonas

aeruginosa infection in such cases can be fatal. The death rate from sepsis can
exceed 80%.
2.3 The application of Choerospondias axillaris on folk drug
On the basis of folk experience, using fire-fighting medicine, for the first
time To Thu used Choerospondias axillaris extract as medicine to cure 18 minor
and severe burns. Fire extinguishing results with simple, cheap, good results: No
multiple, complications, variation, shortened treatment days compared to Western
medicine conditions, comfortable patient, no pain, burns do not sweat. Many cases
can be cured outpatient, at home, very suitable for the conditions of the rural or
remote communities.
At the National Burns Hospital, a number of drugs such as special extract of
the sim, papyrus, yellow papyrus, and especially oval bark with a concentration of
70% tannin have been used to apply burns to patients. With quite popular
properties, many chemical ingredients with good health effects, especially Tannin,

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