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Control of Microbial Growth
Dr. Ashish Jawarkar
Pathologist
Parul Sevashram Hospital
History
Humans vs. Microbes
•
•
•
•
•

infections
diseases
plagues
epidemics
pandemics
Nursery Rhyme

Ring- a - ring of rosies
Pocketful of posies
Achoo ! Achoo !
We all fall down.
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)
Nursery Rhyme
Ring - a - ring of rosies,
A pocketful of posies
Achoo ! Achoo !
We all fall down.
“ I wouldn’t touch it with a 10’ pole”
Humans vs. Microbes
1. Most of History, microbes have been
winning the battle
2. In the last 100 yrs or so the battle has
swung in our favor
• Why?
• Because of our increasing knowledge of how to
Control Microbial Growth
Smallpox

Variola virus
Eradicated in 1977 (Somalia)
Methods to Control Microbial
Growth
1. Physical
2. Chemical
Terms used:
Sterilization vs. Disinfection
Sterilization
• destroying all forms of life

Disinfection
• destroying pathogens or unwanted organisms
Disinfectant vs. Antiseptic
Disinfectant
• antimicrobial agent used on inanimate objects

Antiseptic
• antimicrobial agent used on living tissue
cidal

vs. static

Bactericidal - kills bacteria
Bacteristatic - inhibits bacterial growth
Fungicidal
Fungistatic
Algacidal
Algastatic
Factors that effect Antimicrobial
Activity
1. Temp
2. Time
3. Concentration of Antimicrobial agent
4. Type of Microbe
5. Activity of Microbe
6. Presence of organic matter
Physical Methods of Microbial Control
1. Heat
• works by denaturing enzymes and proteins

A. Thermal Death Point (TDP)
• lowest temp. at which all microorganism in a
liquid culture are killed in 10 minutes

B. Thermal Death Time (TDT)
• minimum length of time in which all
microorganisms in a liquid culture are killed at
a given temperature
Moist Heat
1. Boiling Water
• kills vegetative bacterial cells, Fungi and many
viruses
• not effective for endospores and some viruses
• Hepititis (20 min)
• Some spores may survive boiling water for up
to 20 hrs
Moist Heat
2. Autoclave (Steam under pressure)
• preferred method of sterilization
• Water boils at 100 C
• Increasing the pressure raises the Temp.
• 15 lbs./ per sq. inch (psi) ------> 121 C
•

121 C for 15 min.
Dry Heat
1. Direct Flaming
• Inoculating Loop and Needle 100% effective

2. Incineration
• disposable wastes (paper cups, bags, dressings)

3. Hot Air Sterilization
• Oven ( 170 C for 2 hours)
• used on substances that would be damaged by
moist heat sterilization
• gauzes, dressings or powders
Filtration
Removes microorganisms from solutions
that might be damaged by heat

•
•
•
•

culture media
enzymes
vaccines
antibiotics
Radiation
1. Ionizing Radiation
• gamma rays & x-rays
• penetrates most substances

Used on substances that could be damaged
by heat
•
•
•
•

plastic petri dishes
plastic syringes
catheters
surgical gloves
Radiation
2. Non-Ionizing Radiation
• UV Light
• does not penetrate plastic, glass or proteinaceous
matter

Used to reduce microbial populations
• hospital rooms
• nurseries
• operating rooms

Thymine Dimers
Pasteurization
Disinfection - not sterilization (removes
unwanted organisms)
Mycobacterium tuberculosis
63 C for 30 minutes
72 C for 15 seconds (HTST)
Thermodurics
• able to survive high temps.
Methods used to control Microbial Growth
1. Heat
• Moist Heat
• Boiling Water
• Steam Heat (Autoclave)

• Dry Heat
• Direct Flaming
• Incineration
• Hot Air Sterilization (Oven)

2. Filtration
3. Radiation
• Ionizing Radiation
• Non-Ionizing Radiation

4. Pasteurization (Heat)
Portals of Entry
1. Mucus Membranes
2. Skin
3. Parentarel
1. Mucus Membranes
A. Respiratory Tract
• microbes inhaled into
mouth or nose in
droplets of moisture or
dust particles
• Easiest and most
frequently traveled
portal of entry
Common Diseases contracted via
the Respiratory Tract
Common cold
Flu
Tuberculosis
Whooping cough
Pneumonia
Measles
Strep Throat
Diphtheria
Mucus Membranes
B. Gastrointestinal Tract
• microbes gain entrance thru
contaminated food & water
or fingers & hands
• most microbes that enter the
G.I. Tract are destroyed by
HCL & enzymes of stomach
or bile & enzymes of small
intestine
Common diseases contracted via
the G.I. Tract
Salmonellosis
• Salmonella sp.

Shigellosis
• Shigella sp.

Cholera
• Vibrio cholorea

Ulcers
• Helicobacter pylori

Botulism
• Clostridium botulinum
Fecal - Oral Diseases
These pathogens enter the G.I. Tract at one
end and exit at the other end.
Spread by contaminated hands & fingers or
contaminated food & water
Poor personal hygiene.
Mucus Membranes of the Genitourinary System - STD’s
Gonorrhea
Neisseria gonorrhoeae

Syphilis
Treponema pallidum
Chlamydia
Chlamydia trachomatis
HIV
Herpes Simplex II
Mucus Membranes
D. Conjunctiva –
•

mucus membranes that cover
the eyeball and lines the eyelid

Trachoma
• Chlamydia trachomatis
2nd Portal of Entry: Skin
Skin - the largest organ of the body. When
unbroken is an effective barrier for most
microorganisms.
Some microbes can gain entrance thru
openings in the skin: hair follicles and
sweat glands
3rd Portal of Entry: Parentarel
Microorganisms are deposited into the
tissues below the skin or mucus membranes
Punctures
injections
bites
scratches
surgery
splitting of skin due to swelling or dryness
Preferred Portal of Entry
Just because a pathogen enters your body it
does not mean it’s going to cause disease.
pathogens - preferred portal of entry
Preferred Portal of Entry
Streptococcus pneumoniae
• if inhaled can cause pneumonia
• if enters the G.I. Tract, no disease

Salmonella typhi
• if enters the G.I. Tract can cause Typhoid Fever
• if on skin, no disease

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control of microbial growth

  • 1. Control of Microbial Growth Dr. Ashish Jawarkar Pathologist Parul Sevashram Hospital
  • 3. Nursery Rhyme Ring- a - ring of rosies Pocketful of posies Achoo ! Achoo ! We all fall down.
  • 4. 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
  • 5. Yersinia pestis - Gram (-) bacillus Vectors - Rat and Flea
  • 6. 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
  • 7. 4. Hemorrhaging occurs in lymph nodes, resulting in “black and blue” swellings or Buboes (hence the name Bubonic Plague or Black Death)
  • 8. 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)
  • 9. Nursery Rhyme Ring - a - ring of rosies, A pocketful of posies Achoo ! Achoo ! We all fall down.
  • 10. “ I wouldn’t touch it with a 10’ pole”
  • 11. Humans vs. Microbes 1. Most of History, microbes have been winning the battle 2. In the last 100 yrs or so the battle has swung in our favor • Why? • Because of our increasing knowledge of how to Control Microbial Growth
  • 13. Methods to Control Microbial Growth 1. Physical 2. Chemical
  • 14. Terms used: Sterilization vs. Disinfection Sterilization • destroying all forms of life Disinfection • destroying pathogens or unwanted organisms
  • 15. Disinfectant vs. Antiseptic Disinfectant • antimicrobial agent used on inanimate objects Antiseptic • antimicrobial agent used on living tissue
  • 16. cidal vs. static Bactericidal - kills bacteria Bacteristatic - inhibits bacterial growth Fungicidal Fungistatic Algacidal Algastatic
  • 17. Factors that effect Antimicrobial Activity 1. Temp 2. Time 3. Concentration of Antimicrobial agent 4. Type of Microbe 5. Activity of Microbe 6. Presence of organic matter
  • 18. Physical Methods of Microbial Control 1. Heat • works by denaturing enzymes and proteins A. Thermal Death Point (TDP) • lowest temp. at which all microorganism in a liquid culture are killed in 10 minutes B. Thermal Death Time (TDT) • minimum length of time in which all microorganisms in a liquid culture are killed at a given temperature
  • 19. Moist Heat 1. Boiling Water • kills vegetative bacterial cells, Fungi and many viruses • not effective for endospores and some viruses • Hepititis (20 min) • Some spores may survive boiling water for up to 20 hrs
  • 20. Moist Heat 2. Autoclave (Steam under pressure) • preferred method of sterilization • Water boils at 100 C • Increasing the pressure raises the Temp. • 15 lbs./ per sq. inch (psi) ------> 121 C • 121 C for 15 min.
  • 21. Dry Heat 1. Direct Flaming • Inoculating Loop and Needle 100% effective 2. Incineration • disposable wastes (paper cups, bags, dressings) 3. Hot Air Sterilization • Oven ( 170 C for 2 hours) • used on substances that would be damaged by moist heat sterilization • gauzes, dressings or powders
  • 22. Filtration Removes microorganisms from solutions that might be damaged by heat • • • • culture media enzymes vaccines antibiotics
  • 23. Radiation 1. Ionizing Radiation • gamma rays & x-rays • penetrates most substances Used on substances that could be damaged by heat • • • • plastic petri dishes plastic syringes catheters surgical gloves
  • 24. Radiation 2. Non-Ionizing Radiation • UV Light • does not penetrate plastic, glass or proteinaceous matter Used to reduce microbial populations • hospital rooms • nurseries • operating rooms Thymine Dimers
  • 25. Pasteurization Disinfection - not sterilization (removes unwanted organisms) Mycobacterium tuberculosis 63 C for 30 minutes 72 C for 15 seconds (HTST) Thermodurics • able to survive high temps.
  • 26. Methods used to control Microbial Growth 1. Heat • Moist Heat • Boiling Water • Steam Heat (Autoclave) • Dry Heat • Direct Flaming • Incineration • Hot Air Sterilization (Oven) 2. Filtration 3. Radiation • Ionizing Radiation • Non-Ionizing Radiation 4. Pasteurization (Heat)
  • 27. Portals of Entry 1. Mucus Membranes 2. Skin 3. Parentarel
  • 28. 1. Mucus Membranes A. Respiratory Tract • microbes inhaled into mouth or nose in droplets of moisture or dust particles • Easiest and most frequently traveled portal of entry
  • 29. Common Diseases contracted via the Respiratory Tract Common cold Flu Tuberculosis Whooping cough Pneumonia Measles Strep Throat Diphtheria
  • 30. Mucus Membranes B. Gastrointestinal Tract • microbes gain entrance thru contaminated food & water or fingers & hands • most microbes that enter the G.I. Tract are destroyed by HCL & enzymes of stomach or bile & enzymes of small intestine
  • 31. Common diseases contracted via the G.I. Tract Salmonellosis • Salmonella sp. Shigellosis • Shigella sp. Cholera • Vibrio cholorea Ulcers • Helicobacter pylori Botulism • Clostridium botulinum
  • 32. Fecal - Oral Diseases These pathogens enter the G.I. Tract at one end and exit at the other end. Spread by contaminated hands & fingers or contaminated food & water Poor personal hygiene.
  • 33. Mucus Membranes of the Genitourinary System - STD’s Gonorrhea Neisseria gonorrhoeae Syphilis Treponema pallidum Chlamydia Chlamydia trachomatis HIV Herpes Simplex II
  • 34. Mucus Membranes D. Conjunctiva – • mucus membranes that cover the eyeball and lines the eyelid Trachoma • Chlamydia trachomatis
  • 35. 2nd Portal of Entry: Skin Skin - the largest organ of the body. When unbroken is an effective barrier for most microorganisms. Some microbes can gain entrance thru openings in the skin: hair follicles and sweat glands
  • 36. 3rd Portal of Entry: Parentarel Microorganisms are deposited into the tissues below the skin or mucus membranes Punctures injections bites scratches surgery splitting of skin due to swelling or dryness
  • 37. Preferred Portal of Entry Just because a pathogen enters your body it does not mean it’s going to cause disease. pathogens - preferred portal of entry
  • 38. Preferred Portal of Entry Streptococcus pneumoniae • if inhaled can cause pneumonia • if enters the G.I. Tract, no disease Salmonella typhi • if enters the G.I. Tract can cause Typhoid Fever • if on skin, no disease