microbiology

Medical Micro Lab Exam 2 Study Guide (Exercises 7–14)

This guide keeps everything from your study guide but organizes it so it’s easier to actually memorize for your exam.


I. Laboratory Techniques & General Microbiology



Bacterial Identification Tests

Catalase Test

Purpose:

Detects if bacteria produce the enzyme catalase, which breaks down hydrogen peroxide into water and oxygen.

Positive Result:

Bubbles form

Negative Result:

No bubbles

Used to Differentiate:

  • Staphylococcus = Catalase positive

  • Streptococcus = Catalase negative

Memory Trick:

Staph = Staff = Active = bubbles


Slide Coagulase Test

Purpose:

Detects the enzyme coagulase, which causes plasma to clot.

Positive Result:

Clumping/agglutination

Negative Result:

No clumping

Important:

  • Staphylococcus aureus = positive

  • Staphylococcus epidermidis = negative


Biochemical Tests


Urea Test

Purpose:

Detects urease enzyme that breaks urea into ammonia

Positive:

Pink

Negative:

Yellow/orange

Example Positive:

Proteus


Citrate Test

Purpose:

Shows if bacteria can use citrate as sole carbon source

Positive:

Blue

Negative:

Green


Indole Test

Purpose:

Detects breakdown of tryptophan into indole

Positive:

Red ring after Kovac’s reagent

Negative:

Yellow ring

Example Positive:

E. coli


Methyl Red Test

Purpose:

Detects strong acid production from glucose fermentation

Positive:

Red

Negative:

Yellow


Sugar Tubes

Purpose:

Shows carbohydrate fermentation

Positive:

Yellow = acid produced

Gas Production:

Bubble in Durham tube


Oxidase Test

Purpose:

Detects cytochrome oxidase enzyme

Positive:

Purple/dark blue

Negative:

No color change

Example Positive:

Pseudomonas


TSI Agar (Triple Sugar Iron)

Purpose:

Tests:

  • Glucose

  • Lactose

  • Sucrose

  • Gas

  • H2S production


Reactions

K/A

Red slant / Yellow butt

Only glucose fermented


A/A

Yellow slant / Yellow butt

Glucose + lactose and/or sucrose fermented


K/K

Red slant / Red butt

No sugar fermented


H2S Production

Black precipitate


Antibiotic Sensitivity


Mueller Hinton Agar

Used for antibiotic sensitivity testing

Interpretation:

Measure zone of inhibition

Compare with chart:

  • Sensitive

  • Intermediate

  • Resistant


MIC (Minimum Inhibitory Concentration)

Definition:

Lowest antibiotic concentration that stops visible growth

Example:

First clear broth tube

That = MIC


Environmental & Atmospheric Conditions


Definitions

Aerobic

Needs oxygen

Anaerobic

No oxygen

Microaerophilic

Needs small amount of oxygen

Facultative

Can live with or without oxygen

Capnophilic

Needs extra CO2


N. gonorrhoeae Growth

Needs:

  • GC agar / Chocolate agar

  • CO2 rich environment

  • Warm temp

Important:

Very fragile organism


II. Clinical Applications & Pathogens


Urinary Tract Pathogens

Common:

  • E. coli

  • Proteus

  • Klebsiella

  • Enterococcus

  • Staph saprophyticus

Know selective media appearance


Urine Colony Count Calculation

Given:

1 colony = 1000 bacteria/ml

Formula:

Colonies × 1000 = bacteria/ml

Example:

150 colonies = 150,000 bacteria/ml


Urine Dipstick

Know:

  • Protein

  • Glucose

  • Blood

  • Nitrite

  • Leukocytes

  • pH

  • Ketones

Used for quick urinalysis


Staphylococci


Mannitol Salt Agar (MSA)

Selective For:

Staphylococcus

Differential For:

Mannitol fermentation


S. aureus

Result:

Yellow media

Why:

Ferments mannitol

DNase:

Positive

Coagulase:

Positive


S. epidermidis

Result:

Pink/red media

Why:

Does NOT ferment mannitol

DNase:

Negative

Coagulase:

Negative


Clostridium


Clostridium tetani

Disease:

Tetanus

Characteristics:

  • Anaerobic

  • Gram positive rod

  • Terminal spore

Symptoms:

Lockjaw, muscle spasms


Clostridium perfringens

Disease:

Gas gangrene

Characteristics:

  • Anaerobic

  • Gram positive rod

  • Double zone hemolysis


Meningitis & Specific Pathogens


N. gonorrhoeae

Gram stain:

Gram negative diplococci

Found:

Inside neutrophils

Disease:

Gonorrhea


Treponema pallidum

Seen With:

Dark field microscopy

Disease:

Syphilis


Syphilis Diagnosis

Early Stage:

Dark field microscopy

Later Stage:

Serology (VDRL, RPR, FTA-ABS)


Major Meningitis Pathogens


Neisseria meningitidis

  • Gram negative diplococci

  • Capsule

  • Ferments maltose


Streptococcus pneumoniae

  • Gram positive lancet diplococci

  • Alpha hemolytic

  • Optochin sensitive


Haemophilus influenzae

  • Small gram negative coccobacillus

  • Requires X and V factors

  • Chocolate agar


CSF Changes

Type

Protein

Glucose

WBC

Bacterial

High

Low

High neutrophils

Viral

Slight high

Normal

High lymphocytes

Fungal

High

Low

High lymphocytes

MEMORIZE THIS TABLE


III. Hematology & Blood Analysis


Hematocrit

Definition

Percentage of blood made of RBCs


Increased In:

  • Burns

  • Dehydration


Decreased In:

  • Aplastic anemia

  • Hemolytic anemia


CBC Normal Values

Hemoglobin

Female:
12–16 g/dL

Male:
14–18 g/dL


Hematocrit

Female:
37–47%

Male:
42–52%


White Blood Cells


Neutrophils

Function:

Fight bacteria

Increased In:

Bacterial infections


Lymphocytes

Function:

Fight viruses

Increased In:

Viral infections


Eosinophils

Function:

Parasites + allergies

Increased In:

Parasites + allergic reactions


Monocytes

Function:

Phagocytosis

Become macrophages


Basophils

Function:

Histamine release

Allergic reactions


IV. Vocabulary You MUST Know


Microbiology

Diplococci

Pairs of round bacteria

Agglutination

Visible clumping

Calibrated

Accurately measured

Enterobacteriaceae

Family of gram negative rods

Acid-base indicator

Chemical showing pH change


Pharmacology

Bactericidal

Kills bacteria

Bacteriostatic

Stops bacterial growth

Sensitive

Antibiotic works

Resistant

Antibiotic does not work

Intermediate

May work

Broad spectrum

Works against many bacteria


Blood Terms

Septicemia

Bacteria in blood

Blood culture

Test for bloodstream infection

Erythrocyte

Red blood cell

Leukocyte

White blood cell

Phagocytosis

Cells engulf bacteria

Differential

Percentage of each WBC type


MUST MEMORIZE FIRST (Highest Yield)

TOP PRIORITY:

  1. Catalase vs Coagulase

  2. TSI reactions

  3. MSA results

  4. MIC

  5. CSF table

  6. WBC functions

  7. N. gonorrhoeae / T. pallidum

  8. Hematocrit changes

  9. UTI colony count calculation

  10. Antibiotic sensitivity zones


This is your highest test material.