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Microbiology Class II

Dr. Ashish Jawarkar Consultant Pathologist Parul Sevashram hospital

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Microbiology and the Role of the Medical Assistant

Microbiology study of microorganisms (simple forms of life visible only with a microscope)

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Microbiology and the Role of the Medical Assistant (cont.)

Medical assistant

Assists physician Obtains specimens Prepares specimens for direct examination Prepares specimens for transportation to reference laboratory If office has a POL, performs microbiologic procedures
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How Microorganisms Cause Disease

Cause disease in variety of ways

Use nutrients needed by cells and tissues Damage cells directly Produce toxins

May remain localized or become systemic Transmission

Direct contact Indirect contact

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How Microorganisms Cause Disease (cont.)

Localized symptoms

Generalized symptoms

Swelling Pain Warmth Redness

Fever Tiredness Aches Weakness

Normal flora

Provides a barrier Can cause an infection

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Apply Your Knowledge

1. What role does the medical assistant play in relation to

microbiology? ANSWER: The medical assistant may assist the physician in obtaining specimens, obtain specimens herself, prepare specimens for direct examination or transport to a reference laboratory, and possibly perform microbiologic procedures. 2. How do microorganisms cause disease?

ANSWER: Organisms cause disease by using nutrients needed by cells and tissues, damaging cells directly, or producing toxins.
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Classification of Microorganisms

Structure

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Classification of Microorganisms

Classification by structure

Subcellular DNA or RNA surrounded by a protein coat viruses

Prokaryotic simple cell structure with no nucleus or organelles bacteria

Eukaryotic complex cell structure with nucleus and specialized organelles protozoans, fungi, parasites
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Protozoans single celled eukaryotes Fungi multicelled eukaryotes with cell wall Parasites multicelled eukaryotes without cell wall

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Naming of microorganisms

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Naming of Microorganisms (cont.)

Standardized naming

Genus

Category of biologic classification Example Staphylococcus

Species of organism

Represents a distinct type of microorganisms Examples Staphylococcus aureus and Staphylococcus epidermidis
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Apply Your Knowledge

Describe the classifications of microorganisms and give an example of each.
ANSWER: Microorganisms are classified as: Subcellular organisms that have DNA or RNA surrounded by a protein coat viruses Prokaryotic organisms have a simple cell structure with no nucleus or organelles bacteria Eukaryotic have a complex cell structure with nucleus and specialized organelles protozoans, fungi, parasites

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Viruses

Smallest known infectious agents Subcellular microorganism

Have only nucleic acid surrounded by a protein coat Must live and grow in living cells of other organisms

Hepatitis virus

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Viruses (cont.)

Illnesses caused by viruses

Colds Influenza Croup Hepatitis Warts

AIDS Mumps Rubella Measles Herpes

Vaccines are available for many viruses

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Bacteria

Single-celled prokaryotic organisms Reproduce rapidly Classification

Shape Ability to retain dyes Ability to grow with / without air

Bacillus bacterial classification

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Bacteria: Classification and

Identification

Shape

Coccus spherical, round, or ovoid

Bacillus rod-shaped
Spirillum spiral-shaped Virbrio comma-shaped
Spirillum bacterial classification
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Bacteria: Classification and

Identification

Shape

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Bacteria: Classification and

Identification (cont.)

Ability to retain certain dyes

Grams stain (Gram +ve, Gram ve) Acid-fast stain (AFB+ve, AFB ve)

Ability to grow in presence or absence of air

Aerobes grow best in the presence of oxygen Anaerobes grow best in the absence of oxygen

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Bacteria: Classification and

Identification (cont.)

Special groups

Chlamydiae

Mycobacteria bacilli with a cell wall that differs from most bacteria Rickettsiae

Cell wall structure differs from other bacteria Live and grow within other living cells

Very small Live and grow within other living organisms such as mites and ticks

Mycoplasmas completely lack the rigid cell wall

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Protozoans

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Protozoans

Single-celled eukaryotic organisms, larger than bacteria

Found in soil and water

Illnesses

Malaria Amebic dysentery Trichomoniasis vaginitis

Leading cause of death in developing countries

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Fungi

Eukaryotic organisms with rigid cell wall Yeasts

Single-celled Reproduce by budding

Superficial infections

Athletes foot Ringworm Thrush

Molds

Large, fuzzy, multicelled organisms Produce spores

Can cause systemic infections

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Multicellular Parasites

Organisms that live on or in another organism and use it for nourishment

Parasitic worms

Parasitic insects

Usually due to poor sanitation Roundworms Flatworms Tapeworms

Bite or burrow under the skin Mosquitoes Ticks Lice mites

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Apply Your Knowledge

Matching: ANSWER:
___ D Yeast or mold ___ E Tapeworm / lice A. Virus

___ B Classified by shape

___ A Subcellular organism ___ B May be aerobic or anaerobic A Smallest known organism ___ C Found in soil and water ___

Very C. Protozoan Good!

B. Bacteria D. Fungus E. Multicellular parasite

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How Infections Are Diagnosed

Steps to diagnosis and treatment

1.

Examine the patient

Presumptive diagnosis May or may not need additional tests

2.

Obtain specimen(s)

Label properly Include presumptive diagnosis

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How Infections Are Diagnosed (cont.)

3.

Examine specimen directly

Wet mount Smear Culture medium contains nutrients Examine culture visually and microscopically

4.

Culture specimen

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How Infections Are Diagnosed (cont.)

5.

Determine sensitivity to antibiotics

Treat the patient as ordered

6.

Antimicrobial to kill pathogen or suppress its growth

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Apply Your Knowledge

What is the process for diagnosing an infection?
ANSWER: There are six steps for diagnosis and treatment of an infection:

1. Examine the patient

2. Obtain specimen(s) 3. Examine specimen directly

4. Culture the specimen

5. Determine sensitivity 6. Treat patient / appropriate antimicrobial

Super!

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Specimen Collection

Must be collected correctly

If not, may not grow in culture

Contaminants may be mistakenly identified

Patient may receive incorrect or harmful therapy

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Specimen Collection (cont.)

Devices

Use appropriate collection device or specimen container Sterile swabs absorbent material on the tip

Collection and transporting systems

Sterile, self-contained Transport medium Aerobic or anaerobic

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Specimen Collection: Guidelines

Avoid causing harm, discomfort, or undue embarrassment

Collect from appropriate site Obtain specimen at correct time Use appropriate devices

Obtain sufficient quantity of specimen

Obtain specimen prior to the start of antimicrobial therapy Label correctly

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Specimen Collection (cont.)

Throat culture specimens

Swab back of throat in the area of the tonsils Avoid touching any structures in the mouth Prepare culture plate or prepare correctly for transport to laboratory

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Specimen Collection (cont.)

Urine specimen

Sputum specimen

Clean-catch midstream to minimize contaminants Process within 60 minutes or refrigerate

Specimen from lungs Avoid contaminating specimen with saliva

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Specimen Collection (cont.)

Wound specimen

Stool Specimens

Swab wound or lesion Do not touch outside of wound

Technique varies

Bacterial infection Protozoal or parasitic infection

Instruct patient in correct collection procedure

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Apply Your Knowledge

What are the general guidelines for specimen collection?
ANSWER: They are to avoid causing harm, discomfort, or undue embarrassment; collect from appropriate site; obtain specimen at correct time; use appropriate collection devices; obtain sufficient quantity of specimen; obtain specimen prior to the start of antimicrobial therapy; and label specimen correctly.

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Transporting Specimens to an Outside Laboratory

Many offices send cultures to an outside lab

Three main objectives

Follow proper collection procedures and proper collection device Prevent deterioration of specimen Protect anyone handling specimen
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Direct Examination of Specimens

Enables physician to initiate treatment immediately

Wet mounts

NaCl mixed with specimen of glass slide Presence of pathogen and movement of microorganism

Potassium hydroxide (KOH) mounts

Used if a fungal infection of the skin, nails, or hair is suspected KOH dissolves keratin that can mask presence of a fungus

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Preparation and Examination of Stained Specimens

Quick, tentative diagnosis Differentiation between types of infections

Grams stain

Moderatecomplexity test Bacteria either retain or lose purple color

Gram-positive bacteria Gram-negative bacteria

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Apply Your Knowledge

1. What are the methods for preparing a slide for direct

examination by the physician? ANSWER: They are wet mount and KOH mount.
2. How does the examination of stained specimens

facilitate patient care? ANSWER: Stained specimens enable the physician to provide a quick, tentative diagnosis and differentiate between types of infections.
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Culturing Specimens

Inoculating a culture plate

Transfer some of the specimen onto a culture plate Label the plate correctly Qualitative analysis determination of type of pathogen Quantitative analysis number of bacteria present in sample

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Culturing Specimens (cont.)

Incubating culture plates

35 to 37 C for 24 to 78 hours Agar side up Requires skill and practice Characteristics of colonies Relative number Changes to media around colonies
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Interpreting cultures

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Culturing Specimens (cont.)

Culture media

Liquid, semisolid, or solid forms Contains agar Selective or nonselective

Special culture units

Rapid urine culture Uricult Also available for throat, vaginal, and blood specimens
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Apply Your Knowledge

3. What is the process for culturing a specimen?

ANSWER: The culture medium is inoculated with the specimen and placed in an incubator to promote growth of the organism on the culture medium.

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Determining Antimicrobial Sensitivity

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Determining Antimicrobial Sensitivity

An outside lab reports

Procedure

Sensitive no growth Intermediate little growth Resistant overgrown

Filter paper containing antimicrobial agents placed on inoculated agar plate Incubated for 24 hours Evaluate effectiveness of agent

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Apply Your Knowledge

1. What is the difference between selective and

nonselective culture media?

ANSWER: Selective culture media allows the growth of only certain kinds of bacteria. Unselective culture media support the growth of most organisms.
2. The office received a culture sensitivity report on a

bacteria that said it was resistant to an antimicrobial. What does this mean?
ANSWER: It means that the bacteria was not killed by the antimicrobial and that there was an overgrowth of the bacteria.
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In summary

Types of microorganisms Nomenclature Structure Specimen collection Techniques Antibiotic resistance

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Thank You

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