Medical Microbiology
Department of Microbiology,HMU
PREFACE
of Medical Microbiology
Hong-Xi Gu,Prof.
问题
什么是微生物？
微生物包括哪些种类？
微生物学和医学微生物学
微生物学简史
微生物学现状与远景什么是微生物？
微生物是众多个体微小、结构简单、肉眼不可见的微小生物的总称。
微生物的种类：（三型八大类）
真核细胞型微生物：真菌
原核细胞型微生物：细菌、支原体、立体次体、螺旋体、衣原体、放线菌
非细胞结构型微生物：病毒
微生物与寄生虫的区别
相似：形态微小，需显微镜下观察
区别：微生物以单细胞为单元，没有器官；而寄生虫是多细胞生物，有组织和器官真核细胞型微生物
细胞核的分化程度较高，有核膜、核仁和染色体，胞浆内有完整的细胞器，行有丝分裂。本书只介绍真菌，包括单细胞的酵母和多细胞的霉菌。
原核细胞型微生物
细胞的分化程度较低，仅有原始的核
，无核仁和核膜。
胞浆内无完整的细胞器，如细菌、衣原体、支原体、立克次体、放线菌、
螺旋体。
非细胞结构型微生物
无细胞结构，由核心和蛋白质核壳组成。
核心中只有 RNA或
DNA一种核酸，包括病毒以及结构更简单的亚病毒。
微生物与人类的关系
非常密切：人体内存在大量微生物
有益：工业应用、体内正常菌群（ Vb）
有害：致病，病原微生物（ pathogenic
microbes）。
微生物学与医学微生物学
医学微生物学只研究对人类致病的微生物及其生命活动规律，只涵盖了微生物学的很小一部分医学微生物学发展简史
1600
1700
1800
1900
1677,Observed "little animals"
Antony Leewenhoek (1632-1723)
1796,First scientific Small pox
vaccination
Edward Jenner (1749-1823)
Edward Jenner (1749-1823),after training in London and a period as an army
surgeon,spent his whole career as a country doctor in his native county of
Gloucestershire in the West of England,His research was based on careful case-
studies and clinical observation more than a hundred years before scientists could
explain the viruses themselves,So successful did his innovation prove that by
1840 the British government had banned alternative preventive treatments
against smallpox,"Vaccination," the word Jenner invented for his treatment
(from the Latin vacca,a cow),was adopted by Pasteur for immunization against
any disease,
In the eighteenth century,before Jenner,smallpox was a killer disease,as
widespread as cancer or heart disease in the twentieth century but with the
difference that the majority of its victims were infants and young children,In
1980,as a result of Jenner's discovery,the World Health Assembly officially
declared "the world and its peoples" free from endemic smallpox,
1850,Advocated washing hands
to stop the spread of disease
Ignaz Semmelweis (1818-1865)
Ignaz Semmelweis,a young
Hungarian doctor working in the
obstetrical ward of Vienna General
Hospital in the late 1840s
1861,Disproved spontaneous
generation
Louis Pasteur (1822-1895)
巴氏消毒法
建立炭疽疫苗
建立狂犬病疫苗
1862,Supported Germ Theory of
Disease
Louis Pasteur
1885,First Rabies vaccination
Louis Pasteur,Treatment
and Prevention of Rabies
Louis Pasteur discovered the method for the
attenuation of virulent microorganism that is the
basis of vaccination,He developed vaccines
against chicken cholera,anthrax and swine
erysipelas,After mastering his method of
vaccination,he applied this concept to rabies,
On July 6,1885,Pasteur tested his pioneering
rabies treatment on man for the first time,the
young Joseph Meister was saved,
1867,Practiced antiseptic
surgery
Joseph Lister (1827-1912)
Joseph Lister has contributed much to our
understanding of the causes of disease,He was born in
1827 in Upton,England,He attended London and
Edinburgh Universities,He accepted a position at the
Glasgow Royal Infirmary after graduation,
At the beginning of his career he studied the coagulation
of blood in injuries and surgical wounds,He noted that
there was a very high incidence of infection in wounds,
in spite of efforts to keep rooms clean,In his efforts to
reduce bacteria,he began to spray carbolic acid on
surgical instruments,on wounds,and surgical
dressings,He reduced surgical mortality to 15% by the
year 1860.
His work with antiseptics met with initial resistance in
the medical community but were soon widely accepted,
He was made a baron by Queen Victoria in 1897,Lister
died in England in 1912.
1876,First proof of Germ Theory of
Disease with B,anthracis discovery
Robert Koch (1843-1910)
细菌固定培养法
细菌染色法
发现结核杆菌、炭疽芽胞杆菌、霍乱弧菌
提取旧结素及应用
郭霍法则
The Nobel Prize in Physiology or
Medicine 1905
"for his investigations and
discoveries in relation to
tuberculosis"
1881,Growth of Bacteria on solid media
1882,Outlined Kochs postulates
Robert Koch (1843-1910)
German bacteriologist was the first to cultivate anthrax
bacteria outside the body using blood serum at body
temperature,Building on pasteur's "germ theory",he
subsequently published "Koch's postulates" (1884),the
critical test for the involvement of a microorganism in a
disease,
1,The agent must be present in every case of the disease,
2,The agent must be isolated and cultured in vitro,
3,The disease must be reproduced when a pure culture of the
agent is inoculated into a susceptible host,
4,The agent must be recoverable from the experimentally-
infected host,
This eventually led to,
Development of pure culture techniques
Stains,agar,culture media,petri dishes
1882,Developed acid-fast Stain
Paul Ehrlich (1854-1915)
The Nobel Prize in
Physiology or Medicine
1908
"in recognition of their work
on immunity“
-Shared With Ilya Ilyich Mechnikov,Russian
1884,Developed Gram Stain
1884,Christian Gram
(NOTE,First publication for the Gram stain method)
Gram,C,1884,Ueber die isolirte Farbung der Schizomyceten in Schnitt? und Trockenpraparaten.
REFERENCE,Fortschritte der Medicin,Vol,2,pages 185? 189.
"(I WISH TO THANK HERR DR,RIESS,Director of the General Hospital of Berlin for the facilities and equipment to perform the following studies),
The differential staining method Koch and Ehrlich for tubercle bacilli gives very excellent results either with or without counter-staining,since the bacilli stand out very clearly due to the contrast effect.
It would be very desirable if a similar method for the differential staining of other Schizomycetes were available which could be used routinely by the microscopist.
My studies-as associate of Herr Dr,Friedlander in the morgue of the city hospital in Berlin-have attempted to demonstrate cocci m tlssue sections of lungs of those who have died of pneumonia as well as in experimental animals,As Friedlander has already mentioned briefly in his paper on the micrococci of pneumonia,I
have discovered by experimentation a procedure for the differential staining of pneumococci,In my procedure the nucleus and other tissue elements remain unstained,while the cocci are strongly stained,This makes them much easier to locate than previously,since in ordinary preparations from pneumonia patients,
where such a large amount of exudate occurs,they are impossible to see.
Further studies on the usefulness of this method for other Schizomycetes has gradually shown that this method is an almost general method for all tissue sections and dried preparations....
For staining the ordinary aniline-gentian violet solution of Ehrlich is used,The appropriate sections must be carried up to absolute alcohol and taken from this directly into the dye solution,They should remain in the dye for 1-3 minutes (except tubercle bacilli,which should remain as usual 12-24 hours),Then they are
placed in a solution of iodine-potassium iodide in water (iodine 1.0 part,potassium iodide 2.0 parts,water 300.0 parts) with or without a light rinse with alcohol and allowed to remain there for 1-3 minutes,During this time,a precipitate forms,and the previously dark blue-violet sections now become dark purple-red,
(Footnote,This purple-red color is not soluble in water but dissolves very easily in alcohol,The chemical studies will be continued at a later time.) They are then placed in absolute alcohol until they are completely decolorized,It is well to change thealcohol once or twice during this step,Then the sections are cleared as
usual in clove oil,in which the rest of the dye is dissolved,The nucleus and fundamental tissue is stained only a light yellow (from the iodine) while the Schizomycetes,if any are present in the preparation,are an intense blue (often almost black),The intensity of the staining has not been equaled by any of the current
staining methods,This presents another great advantage of our method,It is possible after the decolorization in alcohol to place the sections for a moment in a weak solution of bismarck brown or vesunne,and then dehydrate again with alcohol,in order toachieve a counterstain,The nucleus will appear brown,while the
Schizomycetes will remain blue.
In this way it is possible to prepare doubly-stained preparations that are just as excellent as those of the tubercle bacillus made after the method of Koch and Ehrlich,Permanent preparations in Canada balsam-xylene or gelatine-glycerol ren~ain unchanged after 4 months.
This method is very quick and easy,The whole procedure takes only a quarter-hour,and the preparations can remain many days in clove oil without the Schizomycete cells becoming decolorized.
It is also useful for dried preparations,It is performed exactly as for tissue sections,except that one treats the cover slip just like a section.
I have tried many times different aniline dyes (rubine-aniline,fuchsinaniline and simple gentian violet solution),but without success,In addition,tincture of iodine or potassium iodide solution,as opposed to iodinepotassium iodide solution,are also ineffective,since the Schizomycetes then are also decolorized,When the
sections are treated with water or dilute alcohol,the results are variable…
I,After iodine treatment,the following forms of Schizomycetes are not decolorized by alcohol.
(a) The coccus of bronchial pneumonia ( 19 cases).
(b) Pyogenic Schizomycetes (9 cases)
(c) Cocci of a liver abscess,,,(1 case)
(d) Cocci and small bacilli in circumscriptive infiltration of the lungs ( 1 case).,,,
(e) Cocci of osteomyelitis (2 cases).
(f) Cocci of suppurative arthritis following scarlet fever ( 1 case)
(g) Cocci of suppurative nephritis following cystitis (3 cases)
(h) Cocci of multiple brain abscesses following empyema (2 cases)
i,Cocci of erysipelas ( 1 case)
(k) Tubercle bacilli (5 cases)
(1) Anthrax bacilli (3 cases) (in mice ),
(m) Putrefactive Schizomycetes (bacilli and cocci)....
II,The following forms of Schizomycetes are decolorized in alcohol after iodine treatment.
(a) 1 case of bronchial pneumonia with cocci that formed cap
(b) 1 case of bronchial pneumonia with cocci that did not form capsules.
(c) Typhoid bacilli (5 cases) are decolorizcd either with or without iodine treatment very easily by alcohol,I have attempted to leave the sections in the dye for as long as 24 hours but without any better results.
Iwould like to make one closing remark,The behavior of the cell nucleus and the Schizomycetes to aniline dyes in other methods are almost identical,whereas with the present staining method a very distinct difference IS visible.
Studies- on Schizomycetes have been significantly improved by the use of this method,It is because of this that I publish my results,although I am well aware that they are brief and with many gaps,It is to be hoped that this method will also be useful in the hands of other workers.
Editor's note,I would like to testify that I have found the Gram method to be one of the best and for many cases the best method which I have ever used for staining Schizomycetes."
1892-1899,Discovered viruses
1982,Dmitri Iosifovich Ivanovski—Tobacco
Mosaic Virus (TMV)
Dmitrii Iwanowski (1892) showed that the infectious agents is smaller than bacteria,He passed the disease sap of TMV through a
Chamberland filter,He inoculated the filtrate to healthy plants and found that the filtrate remained unchanged and reproduced
disease,The concept of filterable infectious agent was against all accepted scientific concepts at that time,so it was not widely
accepted and Iwanowski himself event suspected the disease as a result of a toxin.
1899,Martinus Beijerinck—Recognized viral
dependence on cells for reproduction
In 1898,Martinus Beijerinck confirmed that the causal agent of TMV was indeed filterable and determined that the infectious agent
was able to multiply within living organisms,His research was monumental in that he demonstrated for the first time that TMV was
alive,He hypothesized the agent was liquid,and soluble,and described as contagium vivum fluidum (contagious living fluid),The
way he demonstrated that it was not a toxin was very clever,He took the sap from infected plants,diluted and then inoculated into
healthy plants,Then he repeated the procedure from the newly infected plants,repeated the above procedure,so on and on,At
the end of several transfer,he came to the conclusion that the newly infected plants were as virulent as the original infect ed plants,
If it were a toxin,then it would be have diluted thousand of time,and it would have showed signs of weakening virulence,
1900,Proved mosquitoes carried
the yellow fever agent
Walter Reed
The first human virus that was recognized
1910,Discovered cure for
syphilis
Paul Ehrlich
1928,Discovered Penicillin
Alexander Fleming (1881-1955)
Sir Alexander Fleming Ernst Boris Chain Sir Howard Walter Florey
The Nobel Prize in Physiology or Medicine 1945
"for the discovery of penicillin and its curative effect in various infectious
diseases"
1932,Electronic Microscope
Invented
It was first developed in Germany c.1932;
James Hillier and Albert Prebus,of Canada,
and V,K,Zworykin,of the United States also
made notable contributions to its
development
The scanning electron microscope,
introduced in 1966,gains even greater
resolution by reading the response of the
subject material rather than the direct
reflection of its beam
Using a similar approach,optical scanning microscopes achieve
a resolution of 400 Angstroms,less than the wavelength of the
light being used
Finally,the scanning tunnelling microscope,invented in 1982,
uses not a beam but an electron wave field,which by interacting
with a nearby specimen is capable of imaging individual atoms;
its resolution is an astounding one Angstrom
1963-1970,Hepatitis B Virus
Discovered
1977,Developed a method to
sequence DNA
Paul Berg Walter Gilbert Frederick Sanger
The Nobel Prize in Chemistry 1980
Paul Berg,"for his fundamental studies of the biochemistry of nucleic acids,with
particular regard to recombinant-DNA"
Gilbert & Sanger,"for their contributions concerning the determination of base
sequences in nucleic acids
1980,Small Pox exstinct
World Health Organiszation (WHO) decalred that
Small Pox was extinct on the earth because cow
pox innoculation was widely practised
This was the first time that a disease was declared
to be extinct
1981-1983,AIDS & HIV
In 1981,the first AIDS patient was reported in USA
The etiological agent of AIDS was recognized as
Lymphadenopathy Associated Virus (LAV) by Montagnier,
Pasteur Institute,France in 1983,and Human T-Cell
Lymphotropic Virus Type III (HTLV-III) was recognized by
Gallo,HIT,USA in 1984
LAV and HTLV-III turned out to be the same virus,so was
named as Human Immunodeficiency Virus (HIV)
1983,Polymerase Chain
Reaction invented
Kary Mullis (1944- )
The Nobel Prize in Chemistry 1993
"for contributions to the developments of methods
within DNA-based chemistry"
Kary Mullis,"for his invention of the polymerase chain
reaction (PCR) method"
Michael Smith,"for his fundamental contributions to
the establishment of oligonucleiotide-based,site-
directed mutagenesis and its development for protein
studies"
1995,First microbial genomic
sequence published
Haemophilus Influenza
学习内容和方法
三性两法
生物学特性
致病性
免疫性
微生物学检查法
诊断和特异性防治法
Department of Microbiology,HMU
PREFACE
of Medical Microbiology
Hong-Xi Gu,Prof.
问题
什么是微生物？
微生物包括哪些种类？
微生物学和医学微生物学
微生物学简史
微生物学现状与远景什么是微生物？
微生物是众多个体微小、结构简单、肉眼不可见的微小生物的总称。
微生物的种类：（三型八大类）
真核细胞型微生物：真菌
原核细胞型微生物：细菌、支原体、立体次体、螺旋体、衣原体、放线菌
非细胞结构型微生物：病毒
微生物与寄生虫的区别
相似：形态微小，需显微镜下观察
区别：微生物以单细胞为单元，没有器官；而寄生虫是多细胞生物，有组织和器官真核细胞型微生物
细胞核的分化程度较高，有核膜、核仁和染色体，胞浆内有完整的细胞器，行有丝分裂。本书只介绍真菌，包括单细胞的酵母和多细胞的霉菌。
原核细胞型微生物
细胞的分化程度较低，仅有原始的核
，无核仁和核膜。
胞浆内无完整的细胞器，如细菌、衣原体、支原体、立克次体、放线菌、
螺旋体。
非细胞结构型微生物
无细胞结构，由核心和蛋白质核壳组成。
核心中只有 RNA或
DNA一种核酸，包括病毒以及结构更简单的亚病毒。
微生物与人类的关系
非常密切：人体内存在大量微生物
有益：工业应用、体内正常菌群（ Vb）
有害：致病，病原微生物（ pathogenic
microbes）。
微生物学与医学微生物学
医学微生物学只研究对人类致病的微生物及其生命活动规律，只涵盖了微生物学的很小一部分医学微生物学发展简史
1600
1700
1800
1900
1677,Observed "little animals"
Antony Leewenhoek (1632-1723)
1796,First scientific Small pox
vaccination
Edward Jenner (1749-1823)
Edward Jenner (1749-1823),after training in London and a period as an army
surgeon,spent his whole career as a country doctor in his native county of
Gloucestershire in the West of England,His research was based on careful case-
studies and clinical observation more than a hundred years before scientists could
explain the viruses themselves,So successful did his innovation prove that by
1840 the British government had banned alternative preventive treatments
against smallpox,"Vaccination," the word Jenner invented for his treatment
(from the Latin vacca,a cow),was adopted by Pasteur for immunization against
any disease,
In the eighteenth century,before Jenner,smallpox was a killer disease,as
widespread as cancer or heart disease in the twentieth century but with the
difference that the majority of its victims were infants and young children,In
1980,as a result of Jenner's discovery,the World Health Assembly officially
declared "the world and its peoples" free from endemic smallpox,
1850,Advocated washing hands
to stop the spread of disease
Ignaz Semmelweis (1818-1865)
Ignaz Semmelweis,a young
Hungarian doctor working in the
obstetrical ward of Vienna General
Hospital in the late 1840s
1861,Disproved spontaneous
generation
Louis Pasteur (1822-1895)
巴氏消毒法
建立炭疽疫苗
建立狂犬病疫苗
1862,Supported Germ Theory of
Disease
Louis Pasteur
1885,First Rabies vaccination
Louis Pasteur,Treatment
and Prevention of Rabies
Louis Pasteur discovered the method for the
attenuation of virulent microorganism that is the
basis of vaccination,He developed vaccines
against chicken cholera,anthrax and swine
erysipelas,After mastering his method of
vaccination,he applied this concept to rabies,
On July 6,1885,Pasteur tested his pioneering
rabies treatment on man for the first time,the
young Joseph Meister was saved,
1867,Practiced antiseptic
surgery
Joseph Lister (1827-1912)
Joseph Lister has contributed much to our
understanding of the causes of disease,He was born in
1827 in Upton,England,He attended London and
Edinburgh Universities,He accepted a position at the
Glasgow Royal Infirmary after graduation,
At the beginning of his career he studied the coagulation
of blood in injuries and surgical wounds,He noted that
there was a very high incidence of infection in wounds,
in spite of efforts to keep rooms clean,In his efforts to
reduce bacteria,he began to spray carbolic acid on
surgical instruments,on wounds,and surgical
dressings,He reduced surgical mortality to 15% by the
year 1860.
His work with antiseptics met with initial resistance in
the medical community but were soon widely accepted,
He was made a baron by Queen Victoria in 1897,Lister
died in England in 1912.
1876,First proof of Germ Theory of
Disease with B,anthracis discovery
Robert Koch (1843-1910)
细菌固定培养法
细菌染色法
发现结核杆菌、炭疽芽胞杆菌、霍乱弧菌
提取旧结素及应用
郭霍法则
The Nobel Prize in Physiology or
Medicine 1905
"for his investigations and
discoveries in relation to
tuberculosis"
1881,Growth of Bacteria on solid media
1882,Outlined Kochs postulates
Robert Koch (1843-1910)
German bacteriologist was the first to cultivate anthrax
bacteria outside the body using blood serum at body
temperature,Building on pasteur's "germ theory",he
subsequently published "Koch's postulates" (1884),the
critical test for the involvement of a microorganism in a
disease,
1,The agent must be present in every case of the disease,
2,The agent must be isolated and cultured in vitro,
3,The disease must be reproduced when a pure culture of the
agent is inoculated into a susceptible host,
4,The agent must be recoverable from the experimentally-
infected host,
This eventually led to,
Development of pure culture techniques
Stains,agar,culture media,petri dishes
1882,Developed acid-fast Stain
Paul Ehrlich (1854-1915)
The Nobel Prize in
Physiology or Medicine
1908
"in recognition of their work
on immunity“
-Shared With Ilya Ilyich Mechnikov,Russian
1884,Developed Gram Stain
1884,Christian Gram
(NOTE,First publication for the Gram stain method)
Gram,C,1884,Ueber die isolirte Farbung der Schizomyceten in Schnitt? und Trockenpraparaten.
REFERENCE,Fortschritte der Medicin,Vol,2,pages 185? 189.
"(I WISH TO THANK HERR DR,RIESS,Director of the General Hospital of Berlin for the facilities and equipment to perform the following studies),
The differential staining method Koch and Ehrlich for tubercle bacilli gives very excellent results either with or without counter-staining,since the bacilli stand out very clearly due to the contrast effect.
It would be very desirable if a similar method for the differential staining of other Schizomycetes were available which could be used routinely by the microscopist.
My studies-as associate of Herr Dr,Friedlander in the morgue of the city hospital in Berlin-have attempted to demonstrate cocci m tlssue sections of lungs of those who have died of pneumonia as well as in experimental animals,As Friedlander has already mentioned briefly in his paper on the micrococci of pneumonia,I
have discovered by experimentation a procedure for the differential staining of pneumococci,In my procedure the nucleus and other tissue elements remain unstained,while the cocci are strongly stained,This makes them much easier to locate than previously,since in ordinary preparations from pneumonia patients,
where such a large amount of exudate occurs,they are impossible to see.
Further studies on the usefulness of this method for other Schizomycetes has gradually shown that this method is an almost general method for all tissue sections and dried preparations....
For staining the ordinary aniline-gentian violet solution of Ehrlich is used,The appropriate sections must be carried up to absolute alcohol and taken from this directly into the dye solution,They should remain in the dye for 1-3 minutes (except tubercle bacilli,which should remain as usual 12-24 hours),Then they are
placed in a solution of iodine-potassium iodide in water (iodine 1.0 part,potassium iodide 2.0 parts,water 300.0 parts) with or without a light rinse with alcohol and allowed to remain there for 1-3 minutes,During this time,a precipitate forms,and the previously dark blue-violet sections now become dark purple-red,
(Footnote,This purple-red color is not soluble in water but dissolves very easily in alcohol,The chemical studies will be continued at a later time.) They are then placed in absolute alcohol until they are completely decolorized,It is well to change thealcohol once or twice during this step,Then the sections are cleared as
usual in clove oil,in which the rest of the dye is dissolved,The nucleus and fundamental tissue is stained only a light yellow (from the iodine) while the Schizomycetes,if any are present in the preparation,are an intense blue (often almost black),The intensity of the staining has not been equaled by any of the current
staining methods,This presents another great advantage of our method,It is possible after the decolorization in alcohol to place the sections for a moment in a weak solution of bismarck brown or vesunne,and then dehydrate again with alcohol,in order toachieve a counterstain,The nucleus will appear brown,while the
Schizomycetes will remain blue.
In this way it is possible to prepare doubly-stained preparations that are just as excellent as those of the tubercle bacillus made after the method of Koch and Ehrlich,Permanent preparations in Canada balsam-xylene or gelatine-glycerol ren~ain unchanged after 4 months.
This method is very quick and easy,The whole procedure takes only a quarter-hour,and the preparations can remain many days in clove oil without the Schizomycete cells becoming decolorized.
It is also useful for dried preparations,It is performed exactly as for tissue sections,except that one treats the cover slip just like a section.
I have tried many times different aniline dyes (rubine-aniline,fuchsinaniline and simple gentian violet solution),but without success,In addition,tincture of iodine or potassium iodide solution,as opposed to iodinepotassium iodide solution,are also ineffective,since the Schizomycetes then are also decolorized,When the
sections are treated with water or dilute alcohol,the results are variable…
I,After iodine treatment,the following forms of Schizomycetes are not decolorized by alcohol.
(a) The coccus of bronchial pneumonia ( 19 cases).
(b) Pyogenic Schizomycetes (9 cases)
(c) Cocci of a liver abscess,,,(1 case)
(d) Cocci and small bacilli in circumscriptive infiltration of the lungs ( 1 case).,,,
(e) Cocci of osteomyelitis (2 cases).
(f) Cocci of suppurative arthritis following scarlet fever ( 1 case)
(g) Cocci of suppurative nephritis following cystitis (3 cases)
(h) Cocci of multiple brain abscesses following empyema (2 cases)
i,Cocci of erysipelas ( 1 case)
(k) Tubercle bacilli (5 cases)
(1) Anthrax bacilli (3 cases) (in mice ),
(m) Putrefactive Schizomycetes (bacilli and cocci)....
II,The following forms of Schizomycetes are decolorized in alcohol after iodine treatment.
(a) 1 case of bronchial pneumonia with cocci that formed cap
(b) 1 case of bronchial pneumonia with cocci that did not form capsules.
(c) Typhoid bacilli (5 cases) are decolorizcd either with or without iodine treatment very easily by alcohol,I have attempted to leave the sections in the dye for as long as 24 hours but without any better results.
Iwould like to make one closing remark,The behavior of the cell nucleus and the Schizomycetes to aniline dyes in other methods are almost identical,whereas with the present staining method a very distinct difference IS visible.
Studies- on Schizomycetes have been significantly improved by the use of this method,It is because of this that I publish my results,although I am well aware that they are brief and with many gaps,It is to be hoped that this method will also be useful in the hands of other workers.
Editor's note,I would like to testify that I have found the Gram method to be one of the best and for many cases the best method which I have ever used for staining Schizomycetes."
1892-1899,Discovered viruses
1982,Dmitri Iosifovich Ivanovski—Tobacco
Mosaic Virus (TMV)
Dmitrii Iwanowski (1892) showed that the infectious agents is smaller than bacteria,He passed the disease sap of TMV through a
Chamberland filter,He inoculated the filtrate to healthy plants and found that the filtrate remained unchanged and reproduced
disease,The concept of filterable infectious agent was against all accepted scientific concepts at that time,so it was not widely
accepted and Iwanowski himself event suspected the disease as a result of a toxin.
1899,Martinus Beijerinck—Recognized viral
dependence on cells for reproduction
In 1898,Martinus Beijerinck confirmed that the causal agent of TMV was indeed filterable and determined that the infectious agent
was able to multiply within living organisms,His research was monumental in that he demonstrated for the first time that TMV was
alive,He hypothesized the agent was liquid,and soluble,and described as contagium vivum fluidum (contagious living fluid),The
way he demonstrated that it was not a toxin was very clever,He took the sap from infected plants,diluted and then inoculated into
healthy plants,Then he repeated the procedure from the newly infected plants,repeated the above procedure,so on and on,At
the end of several transfer,he came to the conclusion that the newly infected plants were as virulent as the original infect ed plants,
If it were a toxin,then it would be have diluted thousand of time,and it would have showed signs of weakening virulence,
1900,Proved mosquitoes carried
the yellow fever agent
Walter Reed
The first human virus that was recognized
1910,Discovered cure for
syphilis
Paul Ehrlich
1928,Discovered Penicillin
Alexander Fleming (1881-1955)
Sir Alexander Fleming Ernst Boris Chain Sir Howard Walter Florey
The Nobel Prize in Physiology or Medicine 1945
"for the discovery of penicillin and its curative effect in various infectious
diseases"
1932,Electronic Microscope
Invented
It was first developed in Germany c.1932;
James Hillier and Albert Prebus,of Canada,
and V,K,Zworykin,of the United States also
made notable contributions to its
development
The scanning electron microscope,
introduced in 1966,gains even greater
resolution by reading the response of the
subject material rather than the direct
reflection of its beam
Using a similar approach,optical scanning microscopes achieve
a resolution of 400 Angstroms,less than the wavelength of the
light being used
Finally,the scanning tunnelling microscope,invented in 1982,
uses not a beam but an electron wave field,which by interacting
with a nearby specimen is capable of imaging individual atoms;
its resolution is an astounding one Angstrom
1963-1970,Hepatitis B Virus
Discovered
1977,Developed a method to
sequence DNA
Paul Berg Walter Gilbert Frederick Sanger
The Nobel Prize in Chemistry 1980
Paul Berg,"for his fundamental studies of the biochemistry of nucleic acids,with
particular regard to recombinant-DNA"
Gilbert & Sanger,"for their contributions concerning the determination of base
sequences in nucleic acids
1980,Small Pox exstinct
World Health Organiszation (WHO) decalred that
Small Pox was extinct on the earth because cow
pox innoculation was widely practised
This was the first time that a disease was declared
to be extinct
1981-1983,AIDS & HIV
In 1981,the first AIDS patient was reported in USA
The etiological agent of AIDS was recognized as
Lymphadenopathy Associated Virus (LAV) by Montagnier,
Pasteur Institute,France in 1983,and Human T-Cell
Lymphotropic Virus Type III (HTLV-III) was recognized by
Gallo,HIT,USA in 1984
LAV and HTLV-III turned out to be the same virus,so was
named as Human Immunodeficiency Virus (HIV)
1983,Polymerase Chain
Reaction invented
Kary Mullis (1944- )
The Nobel Prize in Chemistry 1993
"for contributions to the developments of methods
within DNA-based chemistry"
Kary Mullis,"for his invention of the polymerase chain
reaction (PCR) method"
Michael Smith,"for his fundamental contributions to
the establishment of oligonucleiotide-based,site-
directed mutagenesis and its development for protein
studies"
1995,First microbial genomic
sequence published
Haemophilus Influenza
学习内容和方法
三性两法
生物学特性
致病性
免疫性
微生物学检查法
诊断和特异性防治法
