38: Microbiology

Q&A

What is a microorganisms?
微⽣物是什麼 ?

Microorganisms are very small organisms that can only be seen under
the microscope including

protista, algae (unicellular/microscopic), bacteria, cyanobacteria (blue

green bacteria), fungi and viruses.

微⽣物是⾮常細⼩的⽣物，⽤顯微鏡才可看⾒，它們包括原⽣⽣物、⼀些
藻類(單細胞/微⼩
的)、細菌、藍綠細菌(藍綠藻)、真菌和病毒。
What is the structure of a virus?
病毒有什麼的結構 ?

A virus consists of nucleic acids (either DNA or RNA) surrounded by a

protein coat. It has no

nucleus, cytoplasm, cell organelles, cell wall or cell membrane.

病毒含有核酸(只有⼀種，⾮DNA即RNA)，外層包有蛋⽩質外殼，沒有細
胞核、細胞質、細
胞器、細胞壁及細胞膜。
How do viruses multiply?
病毒如何繁殖 ?

38: Microbiology 1
 Viruses can only multiply inside living host cells. The major stages of
multiplication are as follows:

病毒只能在寄主細胞內繁殖，主要的步驟如下:
Attachment 依附:

It adheres to the bacterium by its tail.

它⽤尾部依附於細菌表⾯。
Penetration 注⼊:

The tail sheath contracts. The cell wall of the bacterium is punctured
and the DNA strand of

the phage is injected into the bacterium. The protein coat remains
outside and takes no

further part in the reproductive process.

尾鞘收縮，細菌的細胞壁被刺穿，噬菌體的DNA注⼊細菌體內，蛋⽩
外殼仍然留於
細菌體外，它在隨後的⽣殖作⽤中再沒有功能。
Biosynthesis and replication ⽣化合成與複製:

Phage DNA codes for production of phage enzymes using protein

synthesizing machinery

of host. Phage inactivates and breaks down host DNA and phage
DNA takes over cell

machinery. Phage DNA replicates itself. ie. make new viral proteins
and new viral nucleic acids.

病毒DNA的密碼指令寄主的代謝系統製造病毒酵素，這些酵素可鈍化
和破壞寄主的
DNA，在此，病毒的DNA可完全取代細菌的代謝系統，此後噬菌體進
⾏⾃我複製，
即是製造新病毒蛋⽩質外殼和核酸。
Assembly 組裝:

After about 20 minutes, new phage particles are generated by

assembly of protein coats

38: Microbiology 2
 around phage DNA. 100 to 300 complete phages are formed.

⼆⼗分鐘後，蛋⽩殼和新製成的病毒DNA結合，成為新的病毒粒⼦，
每次⼤約可製
造⼀百到三百個病毒粒⼦。
Release 釋出:

Lysozyme made by phage DNA causes cell lysis releasing phages

to infect other bacterial

hosts if available, or if not, each can remain dormant indefinitely.

稍後病毒DNA會指令製造溶菌酶，將細菌壁溶解，釋出新的病毒粒⼦
去感染其他的
細菌，如沒有適合的寄主時，它可以保持無限期休眠狀態。
State the growth requirement of microorganisms.
列出微⽣物的⽣⻑要求。
(1) Water⽔分

(2) Nutrition營養

(3) Temperature溫度

(4) Oxygen氧氣

(5) Salinity鹽度

(6) pH酸鹼度

(7) Osmotic pressure 滲透壓

(8) Organic growth factors有機⽣⻑因⼦

How do microorganisms grow?

微⽣物如何增⻑ ?

Most microorganisms divide into two identical individuals by binary

fission when they reach a

suitable size. The process repeatd regularly and the population of

microorganisms increases

exponentially in favourable conditions.

當微⽣物達到⼀定體積，⼤部分以⼆分法繁殖，⼀分為⼆，此過程會週期
性地重覆，在有利

38: Microbiology 3
 的環境下，微⽣物會以對數式急劇增⻑。
State the different stages of growth in microorganisms.
列出微⽣物的不同⽣⻑期名稱。
Lag phase遲滯期

Log phase對數期

Stationary phase靜⽌期

Death phase死亡期

Explain lag phase.

解釋遲滯期。
Growth is slow at first, while the microbes acclimate to the food and
nutrients in their new habitat.

They are immature and not yet able to divide.

⽣⻑起初很慢，因為微⽣物都在適應新的⾷物和環境中，他們是不成熟
的，仍未能分裂。
Explain log phase.
解釋對數期
It is the favourable condition for growth, the organisms multiply at a rapid
rate. Most individuals

are reproductive mature and there are plenty of food. Number doubles
every few minutes.

種群在沒有限制因素的情況下，以不斷加快的速率增⻑，因⼤多數個體已
達性成熟，⽽且⾷
物充⾜，數⽬於數分鐘內便可加倍。
Explain stationary phase.
解釋靜⽌期
The population density reaches maximum and becomes constant.
Overcrowding and competition

occur. Nutrient starts to decrease and toxic wastes accumulate.

種群的密度達⾄極限⽽變得穩定，發⽣過度擠迫和競爭，營養開始減少，
有害廢物逐漸積聚。

38: Microbiology 4
 Explain death phase.
解釋死亡期
Toxic waste products build up, food is depleted and the microbes begin
to die. Death rate exceeds

birth rate, resulting in a decrease in number of individuals.

積聚太多的有毒廢物，營養耗盡，微⽣物開始死亡，死亡率⾼於出⽣率，
引致個體數⽬的下降。
Describe the procedures in viable count.
簡述活數的步驟。
(1) Make the appropriate ten-fold serial dilutions

作適當的⼗倍系列稀釋。
(2) Inoculation of the bacteria onto the algar plate.

將細菌接種於瓊脂碟上。
(3) Incubation

溫培。
(4) Counting the cells

數算細胞
(5) Calculation

計算
Describe the procedures in optical density count.
簡述光學密度數的步驟。
A spectrophotometer is used to measure the percentage of light
transmission through a liquid

microbial culture, which is inversely proportional to the population of

microorganisms. The larger

the percentage of light transmission, the smaller is the microbial

population. The population can be

work out by reference to the O.D. population table.

利⽤光譜分析儀量度出穿透過微⽣物培養液光線的百分⽐，這與微⽣物的
數量成反⽐，越多

38: Microbiology 5
 光透過，越少微⽣物，參照光學密度對微⽣物數量表，便可找出微⽣物的
數量。
Describe the procedures in direct count.
簡述直接數的步驟。
The samples are observed under a microscope. By using a cell counter,
the number of bacterial cells from a

given volume of sample is counted, dead cells are also counted too. By
using the known volume trapped in

the grid, the density of the microorganism can be worked out.

將細菌樣本置於顯微鏡下觀察，利⽤細胞數算器，⼀定容量培養液內的細
菌會被數算，包括死細胞。
利⽤已知的網格容量，計算出微⽣物的密度。
Describe the procedures in biomass count.
簡述⽣物量數的步驟。
It involves extracting the microorganisms from the liquid culture and
weighting it directly. The

microorganisms are obtained by getting the residues after centrifugation.

The residues are then

dried by heating at 100oC for 10 hours. The dried residue is weighted

and can represent the

population of the microorganisms.

此法採取從培養液中分離微⽣物然後替他量重，微⽣物可⽤離⼼機抽取，
獲得的殘餘物在
100oC⾼溫下烘乾10⼩時，然後秤量乾燥的殘餘物，這可推算出細菌的數
量。
Explain the importance of aseptic techniques..
解釋消毒技巧的重要性。
Aseptic techniques are the precautionary measures taken to prevent
contamination of pure cultures

and sterile laboratory equipment. We must treat all organisms as

potential pathogens for sake of

38: Microbiology 6
 safety. Many of the organisms can be opportunistic in their abilities to
cause infection.

消毒技巧是防⽌純種培養液及實驗儀器受到污染的預防措施，我們必須將
所有微⽣物當作病
原體處理以策安全，許多微⽣物都可作伺機性感染。
What are aseptic techniques?
何謂消毒技巧 ?

Aseptic techniques are the procedures used to exclude unwanted

microorganisms in order to avoid

contamination. It involves sterilizing equipment and culture media before

and after use for safe

disposal.

消毒技巧是防⽌微⽣物意外外泄，引致污染的必要步驟，它包括在實驗前
或後消毒儀器及培
養基，確保安全。
Describe the method of making solid medium.
簡述固體培養基的製作法。
Boil agar powder / pieces with nutrient solution.

瓊脂⽚加⽔和營養素⼀起煮沸成溶液。
The resulting mixture is poured into sterilized petri dishes. Add cover.

將所得的溶液倒進已消毒的平淺碟內，加上蓋⼦。
The petri dishes with agar solution are sterilized by heating under
pressure in an autoclave.

將盛有瓊脂液的平淺碟提進⾼壓消毒柜，以⾼溫加壓消毒。
After sterilization, let the liquid agar solidifies at room temperature. A
petri dish containing

solidified nutrient agar is called an agar plate. It is ready to culture

bacteria.

消毒後，讓瓊脂液在室溫下冷卻成固體，載有固體營養瓊脂的平淺碟現在
可稱為瓊脂碟，
它已可以⽤來培植細菌。
38: Microbiology 7
 Describe the method of making liquid medium.
簡述液體培養基的製作法。
Dissolve the necessary nutrient in distilled water.

將所需營養素加進蒸餾⽔中。
The resulting mixture (nutrient broth) is poured into sterilized test tubes.

將所得的溶液(營養液體培養基)倒進已消毒的試管內。
The test tubes with nutrient broth are sterilized by heating under
pressure in an autoclave.

將盛有培養液的試管提進⾼壓消毒柜，以⾼溫加壓消毒。
After sterilization, the mouth of test tube is covered with a plug of cotton.
It is ready to culture bacteria.

消毒後，在試管⼝加上棉花塞，它已可以⽤來培植細菌。
Describe the streak plate method in inoculating the culture medium.
簡述接種細菌時的平板劃線法。
The inoculating loop is first sterilized by heating it to red hot.

接種環⾸先加熱⾄通紅以消毒。
The loop is allowed to cool down for 10 seconds.

讓接種環冷卻⼗秒。
The loop is dipped into a broth containing the bacteria.

將接種環浸⼊含有細菌的營養液體培養基內。
The loop is stroked across the surface of agar in a series of continuous
S shaped movements.

把接種環在瓊脂表⾯劃線，劃出多條連續的之字形線條。
The loop is sterilized again by heating, stroked across the surface again
after cool down on

unstroked area.

將接種環再次加熱，冷卻後在瓊脂表⾯未劃線區繼續劃線。
Why should the growing microorganisms be incubated?
為什麼培植中的微⽣物需要溫培 ?

38: Microbiology 8
 An incubator can control the temperature as well as the concentration of
oxygen and carbon dioxide

inside its chamber. This can promote microbial growth.

溫培箱可控制溫度、氧濃度及⼆氧化碳濃度，這可加快微⽣物的⽣⻑。
Compare the advantages and disadvantages of different counting methods.
⽐較不同量度⽅法的優劣。

38: Microbiology 9