Medical Microbiology
 醫學微生物這門課包含的內容繁多, 千頭萬緒. 在 Murray PR
et al. 編寫的 Medical Microbiology 所寫的序言裡說: 學生最
好能 " Thinking like a physician", 要常常問七個問題: Who,
Where, When, Why, Which, What and How.

醫學微生物的內容不只是應用醫學的一項, 它也是一門自然科
學. 它也有科學思辨的趣味, 知識不只是知道名詞而已. 學生還
可以 “ Thinking like a scientist”. 要常常問古早人是怎麼想的?
問題的困難在哪裡? 有沒有比目前的解釋更好的答案?
 知識依自己所長加以斟酌.
 上課內容在網頁上:http://myweb.ncku.edu.tw/~y1357/course/micro.html
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知識之所在!
1. 貴族城堡
2. 公共圖書館
(~2000)
3. 大學(~1000yr)
4. 網域(~10yr)
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一個有用的網址
http://www.scienceprofonline.com/
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http://www.scienceprofonline.com/
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你可以自己學吧!
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如果你可以自己學，
來課堂幹什麼?
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Scale of life：當你看不見的時候，你怎麼辦？
Viroid ~400bp
Paramecium
Bacterial Cell
HIV virion
Prion 27-30 kDa
In mm3
20-50,000
Unicellular
algae
0.01-50
Photosynthetic
bacteria
0.00001-0.01
Pox group
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Discovery of microbes
Antony van Leeuwenhoek
Robert Hooke 1635-?,
My work, which I've done for a
long time, was not pursued in
order to gain the praise I now
enjoy, but chiefly from a craving
after knowledge, which I notice
resides in me more than in most
other men. And therewithal,
whenever I found out anything
remarkable, I have thought it my
duty to put down my discovery on
paper, so that all ingenious people
might be informed thereof.
Antony van Leeuwenhoek. (1632-1723)
Letter of June 12, 1716
他說了一種新態度。
只是看見新東西不能稱之為知識!
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DIY 的年代
http://www.funsci.com/fun3_en/antoni/vlen.htm
資料來源：
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Cell theory
Micrographia; 1664
It takes 190 years
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A short history of microbiology
由看見微生物到認定為致病原花了兩百年

On spontaneous generation: 1668, Francesco Redi (with cover,
against); 1749, John Needham (simple boiling, for); 1765, Lazzaro
Spallanzani (extensive boiling, against)

1673: Anton van Leeuwenhoek (microbes)

1678: Robert Hooke (cell structure)

1861: Louies Pasteur (gooseneck bottle; against)

1870: Joseph Lister (1% carbolic acid, phenol in surgical procedure)

1881: Robert Koch (Koch’s postulate)

1892: Dmitrii Iwanowski (Tobacco Mosaic Virus)

1909: Paul Ehrlich (chemotherapy, arsenicals against spirochetes)

1915: Frederick W. Twort (bacteriophage)

1940: O.T. Avery (DNA as transforming factor)
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光是知道「它的存在」就有很大的威力
Tetanus
Patient number in Canada
After antitoxin vaccine
1941-1995
Tetanus toxin:
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By Robert Kock
In Britain the decline of T.B. was continuous for over 100 years before the introduction of
the vaccine. The department of Health has, surprisingly, decided that routine BCG
vaccination will be phased out in the 1990s. Actually, it comes back due to HIV.
http://www.health.org.nz/tb.html
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看清物質是建立生物學相關學門的基礎：
Microbial classification and structure
Description of bacteria (G+/G-)
Yersinia pestis
Bacillus sp.
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叫不出名字就無法溝通
 Phenotypic classification
 Analytic classification
(composition)
 Genotypic classification
(gene constitute)
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Some key factors
 Improvement of optical
instruments
 Establishment of pure culture
system
 Staining technique
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Big person in microbiology
Robert Koch,1843-1910, Germany
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Communication and classification
By making photomicrobiographs, I can reveal the
bacteria true to nature and free of subjective
misinterpretation
---Rober Koch
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就算是理所當然的事也不容易被接受，為什麼呢？
Now leave everything as it is,
and go to Koch, This man has
made a magnificent discovery
----Julius Cohnheim 1817-1891
Whole business seemed quite
improbable; Anything I couldn’t
see with a dry lens wasn’t worth
looking at (知識轉向是科學哲學的議題).
----Rudolf Virchow 1821-1902
Koch’s demonstrations in the Institute of plant
pathology; University of Breslau (15 Nov. 1876)
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Morphological study of bacteria

Instruments:

Light microscope: 0.2 mm


Electron microscope: 0.0001 mm


Phase-contrast; dark field
TEM; SEM
Near-field microscope: atom-level
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Growth/Culture condition
Plating technique of Robert Koch for
single colony isolation
Microbiol. Mol. Biol. Rev. 64:847-867 (2000)
The majority of microbes persist
attached to surfaces within a
structured biofilm ecosystem and not
as free floating organisms.
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Classification

Phenotypic classification


Analytic classification


Morphology; Biotyping; Serotyping; phage typing;
Antibiogram….
Cell wall fatty acid; lipid; protein; enzyme
isomers…..
Genotypic classification

G/C ratio; DNA sequence compare; DNA/RNA
hybridization, plasmid….
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Morphological study

Unstained


General shape, motility, spore, intracellular granules
Fix and stained


Simple stain:

Positive: methylene blue, methyl violet, basic fuchsin,
silver…

Negative: acidic dye for background…
Gram’s stain: (by Hans C. Gram, 1884) base on the
resistance to decoloration with acetone, alcohol or aniline oil
after stained with a triphenyl methane dye. (+: purple; -:red)

Acidfast stain reaction: (Ziehl-Neelsen method) cell wall
consists of lipids and forms a special anatomical disposition of
the lipids.

Immunostaining: labeled antibodies
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For lecture only
簡易二分法
Simple Stain
Cocci
Bacilli
Gram Stain
Gram Stain
Gram
Negative
cocci
Gram
positive
cocci
Gram positive bacilli
Gram
negative
bacilli
Acid Fast stain
MacConkey’s
Acid Fast
Mannitol Salt
No
color
change
yellow
Not
acid
Mycobacterium fast
Pink
colonies tuberculosis
Endospore stain
pink
Salmonella
pullorum
Staphylococcus Staphylococcus
aureus
epidermidis
E. coli
Enterobacter
aerogenes
Forms
endospores
Bacillussubtilus
The prokaryotes and eukaryotic protists were more
difficult to classify. They are much simpler, and there are
few obvious features.
Classifications based upon features such as these were
not natural systems (Bergey's Manual of Determinative
Bacteriology )










Gram stain (cell wall structure)
Mole percent G+C in the genome
Growth temperature
Ability to form heat stable spores
Electron acceptors for respiration
Photosynthetic ability
Motility
Cell shape
Ability to use various carbon and nitrogen
sources
Special nutritional requirements
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For lecture only
Classification: Tree of evolution
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在分類學之外插曲
• 一八九六年，台灣安平爆發黑死病。年底，緒方正規應邀到
台灣研究，在台北小南門外設立研究室。一八九七年初，他
發現人、鼠均由老鼠身上的跳蚤傳染病菌。
• 一八九八年，巴斯德研究所的席蒙（Paul Louis Simond,
1858-1947）到印度孟買研究黑死病，以實驗證實了跳蚤是
病菌傳播者。從此，黑死病又稱為鼠疫。
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北里柴三郎
Robert Koch
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科學發展2003年7月，367期76-79
•
一九○一年，北里與助手中川合寫的一篇文章說道：黑死病菌是桿菌，但是這種桿菌
在腫大的淋巴結裡發現得最多，在肺、心、腦、脊髓中，牠們往往看來像球菌。在培
養基生長的菌落，以球菌形為主。但是仔細觀察後，就會發現牠們是串接在一起的桿
菌，而不是球菌團；在血清中培養的，呈典型桿菌狀。這種細菌是格蘭氏陽性菌。可
是，葉赫森一開始就明白地報告：他發現的細菌是格蘭氏陰性的。
•
現在鼠疫菌的學名，既不是北里的「鼠疫桿菌」，也不是葉赫森的「巴斯德鼠疫菌」
或勞森的「北里桿菌」， 而是「葉赫森屬鼠疫種」（Yersinia pestis），簡稱葉赫森菌
。那麼，北里柴三郎發現的細菌究竟是打哪兒來的，他真的搞錯了嗎？根據細菌學者
的意見，北里的第一篇正式報告大體而言是正確的，最引人矚目的問題出在他說「鼠
疫菌有一些呈格蘭氏陽性反應」，不免讓人對於他的正確描述也看輕了。
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總督府民政長官後藤新平直言要以醫療作為政治的後盾，
在生物學為基礎的管理政策之下，採用醫師來代替歐美
的傳教士，設置公醫制度，以宣傳「先進的日本」。
•日本進駐台灣後，隔一年，先進的日本科學宣傳馬上遭到黑死病的考驗。
1896年，日軍駐台南部隊二等軍醫村上彌穗若從病人腺腫液內培養出細
菌，檢體送往東京軍醫學校，經教官岡田國太郎判定為葉爾辛鼠疫桿菌
(Yersinia pestis)。同年12月，東京帝國大學教授緒方正規、山極勝三郎
在台北小南門外設立了鼠疫研究室，由堀內次雄輔佐，進行細菌病理和
臨床研究。他們証實細菌Y. pestis感染老鼠和人導致鼠疫，而跳蚤是傳播
媒介。
•台灣鼠疫流行疫期直至1917年才告平息，22年的流行期間，官方正式的
統計資料記載病患人數總計30,101人，其中 24,104人死亡，死亡率達百
分之八十。其他各地各時期統計的死亡率約在百分之六十至九十之間。
在高死亡率的恐慌之下，疾病的治療及控制成了各方角力的大事。
•日本醫生所採用的血清療法，與當時台灣人所熟悉的漢藥完全不同。它
起源是在疾病病原論的認知之下，由日本細菌學者北里柴三郎跟德國細
菌學者Emil Adolf von Behring (1854-1917)於1890年在科霍 (Robert
Koch) 的實驗室中研發白喉毒素的抗毒血清，宣稱血清能對抗細菌毒素。
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楊倍昌《科學之美：生物科學史閱讀手記》2012
日治時期台灣說理方式的轉向
計算錯誤
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明治三十年
明治三十二 明治三十二年台灣
年
醫生
鼠疫患者總
數
210人
235人
890人
痊癒
83人
108人
391人
死亡
127人
127人
599人*/499人
死亡率 (%)
60.48
54.04
67.3*/56.06
雖然日本公醫的醫療成效無法深入一般台灣民眾，但是在台灣總督府報告的檔
案中，他們以漢醫不曾使用的新方法紀錄了台灣的醫療成績。它包括以實驗室
為基礎的病理檢查(顯微鏡觀察病人淋巴腺中的細菌數目)、規劃對照組，數量
化的登錄格式。雖然血清治療法成效不彰，而且築山揆一取樣的適切性和對照
組的規劃仍有討論的餘地，但是這種以實驗室支援的醫學檢驗，以及療效的報
告方式，呈現出舊台灣文化之中不曾有的思維理路。
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