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Quiz 2

Dr. Ashish Jawarkar
M.D. Path
Parul Sevashram Hospital
Q-1
 Which class of organisms causes the

disease in picture
Athlete’s foot
 A. Bacteria
 B. Fungi
 C. Virus
 D. Protozoa
Fungi
 Eukaryotic organisms

with rigid cell wall
 Yeasts



Single-celled
Reproduce by budding

 Superficial infections




Athlete’s foot
Ringworm
Thrush

 Molds




Large, fuzzy,
multicelled organisms
Produce spores

 Can cause systemic

infections
Quiz 2
Quiz 2
Q-2
 Which of the following is semipermeable
 A. cell membrane
 B. cell wall
 C. Nucleus
 D. Ribosomes


Cytoplasmic (Plasma)
Thin membrane
layer 5-10 nm, separates cell wall from cytoplasm



Acts as a semipermeable membrane: controls the
inflow and outflow of metabolites



Composed of lipoproteins with small amounts of
carbohydrates
Quiz 2
Q-3
 What are the contents of Mac Conkey’s

medium?
 A. Peptone
 B. Lactose
 C. Neutral Red
 D. All of the above
Differential media
 A media which has substances incorporated in it
enabling it to distinguish between bacteria.
 Eg: Mac Conkey’s medium






Peptone
Lactose
Agar
Neutral red
Taurocholate

 Distinguish between lactose fermenters & non

lactose fermenters.
 Lactose fermenters – Pink colonies
 Non lactose fermenters – colourless colonies
Q-4
 What is this used for?
 For removing oxygen, to grow anaerobic

bacteria
ANAEROBIC CULTURE METHODS
 Anaerobic bacteria differ in their requirement

and sensitivity to oxygen.
 Cl.tetani is a strict anaerobe – grows at an
oxygen tension < 2 mm Hg.

Methods:
Production of vacuum
 Displacement of oxygen with other gases
 Chemical method
 Biological method
 Reduction of medium

Production of vacuum:
 Incubate the cultures in a vacuum desiccator.
Displacement of oxygen with other gases
 Displacement of oxygen with hydrogen, nitrogen,
helium or CO2.
 Eg: Candle jar
Quiz 2
Chemical method
 Alkaline pyrogallol absorbs oxygen.

McIntosh – Fildes’ anaerobic jar
 Consists of a metal jar or glass jar with a metal

lid which can be clamped air tight.
 The lid has 2 tubes – gas inlet and gas outlet
 The lid has two terminals – connected to
electrical supply.
 Under the lid – small grooved porcelain spool,
wrapped with a layer of palladinised asbestos.
Quiz 2
Working:
 Inoculated plates are placed inside the jar and
the lid clamped air tight.
 The outlet tube is connected to a vacuum pump
and the air inside is evacuated.
 The outlet tap is then closed and the inlet tube is
connected to a hydrogen supply.
 After the jar is filled with hydrogen, the electric
terminals are connected to a current supply, so
that the palladinised asbestos is heated.
 Act as a catalyst for the combination of hydrogen
with residual oxygen.
Gaspak
 Commercially available disposable envelope.
 Contains chemicals which generate H2 and CO2 on
addition of water.
 Cold catalyst – in the envelope
 Indicator is used – reduced methylene blue.



Colourless – anaerobically
Blue colour – on exposure to oxygen
Quiz 2
Q-5
 What is resolution
 A. the ability of the microscope to

enlarge the object
 B. the ability of the microscope to show
two nearby placed objects separately
Important Vocabulary :
magnification  mag-ne-fe-'ka-shen n 1.
apparent enlargement of an object 2. the
ratio of image size to actual size
A magnification of "100x" means that the
image is 100 times bigger than the actual
object.
resolution  rez-e-loo-shen n 1. clarity,
sharpness 2. the ability of a microscope to
show two very close points separately
Q-6
 Which type of microscope has been

used for taking this picture?
 A. Bright field microscope
 B. Dark field microscope
 C. Phase contrast microscope
 D. Fluorosence microscope
Bright-field Microscope
 Contains two lens systems for magnifying

specimens
 Specimens illuminated directly from above or
below
 Advantages: convenient, relatively
inexpensive, available
 Disadvantages: R.P 0.2 µm at best; can
recognize cells but not fine details
 Needs contrast. Easiest way to view cells is
to fix and stain.
Different magnifications
Special Microscopy
Applications
 Dark Field
 Phase Contrast
 Fluorescence
 Electron Microscope
Dark Field Microscopy
 special condenser

diaphragm

occludes direct light,
passes wide angle
light
 angle too wide to
enter objective
diffracted light scattered
enters objective
objects light on dark background


diffracted
light
Phase Contrast Microscopy
 light rays through objects of different η → change in

phase, not intensity
 special ring-shaped condenser diaphragm
 special glass disc in objective



change phase differences to intensity differences
can view transparent
objects as dark on light
background (without staining)

 Right; human brain glial

cells
Fluorescence Microscopy
 Illuminate specimen with UV → visible fluorescence

(filter removes harmful UV)
 View auto-fluorescent objects (e.g., chloroplasts)
 Stain with specific fluorescent dyes, which absorb in
region 230-350 nm & emit orange, yellow or greenish
light
 Images appear coloured against a dark background
Electron Microscopy
Q-7
 Steps of staining include
 1. primary stain
 2. decolorisation
 3. counter stain

Which dyes are used for these steps?
In gram staining and acid fast staining
Primary
staining

Decolorisation

Counter
staining
Primary stain

Decolorisation

Counter stain
Q-8
 What is sterilisation?
 A. Killing all organisms on the object
 B. Removing only pathogenic organisms
DEFINITION
STERILIZATION
The process of freeing an article from
microorganisms including their spores.
DISINFECTION:
Reducing the number of pathogenic
microorganisms to the point where
they no longer cause diseases.
Q-9
 Which is true?
 A. Bacteriostatic agent doesnot kill

bacteria
 B. Bactericidal agent kills bacteria
 C. All of the above
Bacteriostatic Agent:
An agent that inhibits the growth of bacteria, but
does not necessarily kill them.
Bactericide:
An agent that kills bacteria. Most do not kill
Endospores.

Sporicide:
An agent that kills spores.
Q-10
 Which method is used to sterilize?
 A. Hot air oven
 B. Autoclave
 C. Pasteurization
 D. Ethylene oxide gas
Q-11
 Which organism causes plague
 A. streptococcus pneumoniae
 B. mycobacterium tuberculosis
 C. vibrio cholerae
 D. yersinia pestis
Bubonic Plague or the Black
Death
 Epidemic swept thru Europe in the Middle

Ages (13th and 14th centuries)
 40 million people were killed


About 1/3 of the population of the continent

 Etiological agent:


Yersinia pestis

 2 Vectors



Rat
Flea

Gram (-) rod
Yersinia pestis - Gram (-) bacillus
Vectors - Rat and Flea
Bubonic Plague Infection
 1. Flea bite with Yersinia pestis
 2. Bacteria multiply in the bloodstream


Bacteremia

 3. Bacteria localize in lymph nodes,

especially axillary and groin areas
4. Hemorrhaging occurs in lymph nodes, resulting in
“black and blue” swellings or Buboes (hence the name
Bubonic Plague or Black Death)
Bubonic Plague Infection
 5. If untreated, about 50 % Mortality Rate
 6. If bacteria spread to the lungs, it becomes

Pneumonic Plague and is now highly
contagious (Almost a 99 % Mortality Rate)
Quiz 2

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Quiz 2

  • 1. Quiz 2 Dr. Ashish Jawarkar M.D. Path Parul Sevashram Hospital
  • 2. Q-1  Which class of organisms causes the disease in picture
  • 4.  A. Bacteria  B. Fungi  C. Virus  D. Protozoa
  • 5. Fungi  Eukaryotic organisms with rigid cell wall  Yeasts   Single-celled Reproduce by budding  Superficial infections    Athlete’s foot Ringworm Thrush  Molds   Large, fuzzy, multicelled organisms Produce spores  Can cause systemic infections
  • 8. Q-2  Which of the following is semipermeable  A. cell membrane  B. cell wall  C. Nucleus  D. Ribosomes
  • 9.  Cytoplasmic (Plasma) Thin membrane layer 5-10 nm, separates cell wall from cytoplasm  Acts as a semipermeable membrane: controls the inflow and outflow of metabolites  Composed of lipoproteins with small amounts of carbohydrates
  • 11. Q-3  What are the contents of Mac Conkey’s medium?  A. Peptone  B. Lactose  C. Neutral Red  D. All of the above
  • 12. Differential media  A media which has substances incorporated in it enabling it to distinguish between bacteria.  Eg: Mac Conkey’s medium      Peptone Lactose Agar Neutral red Taurocholate  Distinguish between lactose fermenters & non lactose fermenters.
  • 13.  Lactose fermenters – Pink colonies  Non lactose fermenters – colourless colonies
  • 14. Q-4  What is this used for?
  • 15.  For removing oxygen, to grow anaerobic bacteria
  • 16. ANAEROBIC CULTURE METHODS  Anaerobic bacteria differ in their requirement and sensitivity to oxygen.  Cl.tetani is a strict anaerobe – grows at an oxygen tension < 2 mm Hg. Methods: Production of vacuum  Displacement of oxygen with other gases  Chemical method  Biological method  Reduction of medium 
  • 17. Production of vacuum:  Incubate the cultures in a vacuum desiccator. Displacement of oxygen with other gases  Displacement of oxygen with hydrogen, nitrogen, helium or CO2.  Eg: Candle jar
  • 19. Chemical method  Alkaline pyrogallol absorbs oxygen. McIntosh – Fildes’ anaerobic jar  Consists of a metal jar or glass jar with a metal lid which can be clamped air tight.  The lid has 2 tubes – gas inlet and gas outlet  The lid has two terminals – connected to electrical supply.  Under the lid – small grooved porcelain spool, wrapped with a layer of palladinised asbestos.
  • 21. Working:  Inoculated plates are placed inside the jar and the lid clamped air tight.  The outlet tube is connected to a vacuum pump and the air inside is evacuated.  The outlet tap is then closed and the inlet tube is connected to a hydrogen supply.  After the jar is filled with hydrogen, the electric terminals are connected to a current supply, so that the palladinised asbestos is heated.  Act as a catalyst for the combination of hydrogen with residual oxygen.
  • 22. Gaspak  Commercially available disposable envelope.  Contains chemicals which generate H2 and CO2 on addition of water.  Cold catalyst – in the envelope  Indicator is used – reduced methylene blue.   Colourless – anaerobically Blue colour – on exposure to oxygen
  • 24. Q-5  What is resolution  A. the ability of the microscope to enlarge the object  B. the ability of the microscope to show two nearby placed objects separately
  • 25. Important Vocabulary : magnification  mag-ne-fe-'ka-shen n 1. apparent enlargement of an object 2. the ratio of image size to actual size A magnification of "100x" means that the image is 100 times bigger than the actual object. resolution  rez-e-loo-shen n 1. clarity, sharpness 2. the ability of a microscope to show two very close points separately
  • 26. Q-6  Which type of microscope has been used for taking this picture?
  • 27.  A. Bright field microscope  B. Dark field microscope  C. Phase contrast microscope  D. Fluorosence microscope
  • 28. Bright-field Microscope  Contains two lens systems for magnifying specimens  Specimens illuminated directly from above or below  Advantages: convenient, relatively inexpensive, available  Disadvantages: R.P 0.2 µm at best; can recognize cells but not fine details  Needs contrast. Easiest way to view cells is to fix and stain.
  • 30. Special Microscopy Applications  Dark Field  Phase Contrast  Fluorescence  Electron Microscope
  • 31. Dark Field Microscopy  special condenser diaphragm occludes direct light, passes wide angle light  angle too wide to enter objective diffracted light scattered enters objective objects light on dark background  diffracted light
  • 32. Phase Contrast Microscopy  light rays through objects of different η → change in phase, not intensity  special ring-shaped condenser diaphragm  special glass disc in objective   change phase differences to intensity differences can view transparent objects as dark on light background (without staining)  Right; human brain glial cells
  • 33. Fluorescence Microscopy  Illuminate specimen with UV → visible fluorescence (filter removes harmful UV)  View auto-fluorescent objects (e.g., chloroplasts)  Stain with specific fluorescent dyes, which absorb in region 230-350 nm & emit orange, yellow or greenish light  Images appear coloured against a dark background
  • 35. Q-7  Steps of staining include  1. primary stain  2. decolorisation  3. counter stain Which dyes are used for these steps? In gram staining and acid fast staining
  • 38. Q-8  What is sterilisation?  A. Killing all organisms on the object  B. Removing only pathogenic organisms
  • 39. DEFINITION STERILIZATION The process of freeing an article from microorganisms including their spores.
  • 40. DISINFECTION: Reducing the number of pathogenic microorganisms to the point where they no longer cause diseases.
  • 41. Q-9  Which is true?  A. Bacteriostatic agent doesnot kill bacteria  B. Bactericidal agent kills bacteria  C. All of the above
  • 42. Bacteriostatic Agent: An agent that inhibits the growth of bacteria, but does not necessarily kill them.
  • 43. Bactericide: An agent that kills bacteria. Most do not kill Endospores. Sporicide: An agent that kills spores.
  • 44. Q-10  Which method is used to sterilize?
  • 45.  A. Hot air oven  B. Autoclave  C. Pasteurization  D. Ethylene oxide gas
  • 46. Q-11  Which organism causes plague  A. streptococcus pneumoniae  B. mycobacterium tuberculosis  C. vibrio cholerae  D. yersinia pestis
  • 47. Bubonic Plague or the Black Death  Epidemic swept thru Europe in the Middle Ages (13th and 14th centuries)  40 million people were killed  About 1/3 of the population of the continent  Etiological agent:  Yersinia pestis  2 Vectors   Rat Flea Gram (-) rod
  • 48. Yersinia pestis - Gram (-) bacillus Vectors - Rat and Flea
  • 49. Bubonic Plague Infection  1. Flea bite with Yersinia pestis  2. Bacteria multiply in the bloodstream  Bacteremia  3. Bacteria localize in lymph nodes, especially axillary and groin areas
  • 50. 4. Hemorrhaging occurs in lymph nodes, resulting in “black and blue” swellings or Buboes (hence the name Bubonic Plague or Black Death)
  • 51. Bubonic Plague Infection  5. If untreated, about 50 % Mortality Rate  6. If bacteria spread to the lungs, it becomes Pneumonic Plague and is now highly contagious (Almost a 99 % Mortality Rate)

Hinweis der Redaktion

  1. Uses of darkfield and Phase contrast microscopy View unstained cells Not subject to shrinkage, artefacts Some bacteria do not stain easily, e.g., spirochetes (Borrelia, Leptospira, Treponema) View living cells Can observe processes, e.g., motility, predation