SIMULATED NO.

GENERAL RULE:

1. ALL cocci are gram positive (+) Except:

Neisseria
Branhamella
Veillonella
2. ALL bacilli are gram negative (-) Except:
Bacillus
Clostridium
Coynebacterium
Erysipelothrix
Listeria
Kurthia
Mycobacteria
Rothia
Fungus-like: Actinomyces, Nocardia
3. SPIRAL are gram negative (-)
*they are hard to stain
4. Mycoplasma and Ureaplasma are gram negative (-)

AEROBE
COCCI BACILLI
GRAM POSITIVE GRAM NEGATIVE GRAM POSITIVE GRAM NEGATIVE
Staphylococcus Neiserria Bacillus Acinetobater
Streptococcus Branhamella Corynebacterium Aeromonas
Micrococcus Erysipelothrix Alcaligenes
Lactobacillus Bordetella
Listeria Brucella
Mycobacterium Enterobacteriacea
Nocardia Francisella
Legionella
Pseudomonas
Vibrio
ANAEROBE
COCCI BACILLI
GRAM POSITIVE GRAM NEGATIVE GRAM POSITIVE GRAM NEGATIVE
Peptococcus Veillonella Actinomyces Bacteroides
Peptostreptococcus Clostridium Fusobactrium
Sarcina Propionobacterium

Haemophilus

 Nonmotile, nonsporefoming
 Facultative anaerobe
 Most are oxidase and catalase positive
 Preferred incubation at 35-37ᵒ C

SPECIES REQUIREMENTS BETA HEMOLYSIS D-ALA

X FACTOR V FACTOR
H. influenza √ √ - -
H. parainfluenzae X √ - +
H. hemolyticus √ √ + -
H. parahemolyticus X √ + +
H. aegypticus √ √ - -
H. aprophilus X X - +
H. paraphrophilus X √ - +
H. ducreyi X x - -
H. influenza (Pfeiffer’s bacillus)

 Six serotypes (a,b,c,d,e and f); most frequently encountered serotype in infection is b
 Encapsulated strain are pathogenic
 Main cause of meningitis in children <years old
 Associated with respiratory condition including epiglottitis

H.aegypticus

H. ducreyi

 Infective agent of chancoid/soft chancre, venereal disease characterized by painful ulcers in the genitalia
 Direct examination short bacilli in a school of fish arrangement.
 Smallest pathogenic bacilli (largest B.anthracis)

HEMOGLOBIN

 1 g Hb can carry 1.34 mL of O2

 1 g Hb carries a constant 3.47 mg iron

MAJOR ENDOCRINE GLANDS AND THEIR HORMONES

ENDOCRINE HORMONE FUNCTION HYPOSECRETION HYPERSECRETION

GLAND
Anterior Pituitary Growth hormone Major effects are directed to Hyposecretion during Hypersecretion produces
Gland (GH) the growth of skeletal childhood leads to gigantism (childhood) and
(adenohypophysis) Somatotropin muscles and long bones of pituitary dwarfism acromegaly (adulthood)
the body
Most abundant
hormone of anterior
pituitary
Prolactin (PRL) Stimulates production of
breast milk
Adenocorticotrophi Stimulates adrenal cortex
c to release its hormones
Hormone (ACTH)
Thyrothropic Stimulates the thyroid gland
hormone (TH) to release thyroid hormones

Thyroid-stimulating
hormone (TSH)
Follicle-stimulating Beginning at puberty, Sterility in both male
hormone (FSH) stimulates follicle and female
development and estrogen
production by female
ovaries; promotes sperm
production in males
Luteinizing Beginning at puberty, Sterility in both male
hormones (LH) stimulates ovulation, and female
converts the ruptured ovarian
follicle to a corpus luteum to
produce progesterone;
stimulates male testes to
produce testosterone
Posterior Pituitary Oxytocin Stimulates powerful uterine
Gland Released in contractions and causes milk
(neurohypophysis) significant amount ejection in nursing women
only during
childbirth and in
 nursing women
Antidiuretic hormone Causes kidney tubule to Diabetes insipidus
(ADH) reabsorb and conserve body
water and increases blood
Vasopressin pressure by constricting
anterioles
Thyroid gland Thyroxine (T4) Body’s metabolic hormone. Hyposecretion of Hypersecretion results in
It increases the rate at which thyroxine results in Grave’s disease and other
Triiodothyronine cells oxidize glucose and is cretinism (children) forms of hyperthyroidism
(T3) necessary for normal growth
and development
Calcitonin Causes calcium to be
deposited in long bones
Parathyroid Gland Parathyroid Causes bone calcium to be result to tetany leads to exteme bone
hormone (PTH) liberated to the blood wasting and fractures
Adrenal Cortex Mineralocorticoids Regulate sodium and A generalized Hypersecretion of
potassium ion reabsorption hypoactivity of adrenal cortex
mainly aldosterone by the kidneys. Their adrenal cortex leads to hormones can result in
release is primarily Addison’s disease Hyperaldosteronism,
(outermost – zona stimulated by low Na+/high Cushing’s disease and/or
glomerulosa) K+ levels in the blood masculinization

Glucocorticoid Enable the body to resist

which include long-term stress by
cortisone and increasing blood glucose
cortisol levels and decreasing the
inflammatory response.
(middle – zona
fasciculata)
Sex hormones
Androgens (male)
w/ some estrogen
(female)

(innermost – zona
reticularis)
Adrenal Medulla Catecholamines : Hypersecretion leads to
Epinephrines symptoms typical of
(adrenaline) sympathetic nervous
activity
Norepinephrine
(noradrenaline)
Islets of Langerhans Insulin Increase the rate of glucose Diabetes mellitus
of pancreas uptake and metabolism by
By beta cells body cells
Glucagon Stimulates the liver to release
glucose to blood, thus
By alpha cells increasing blood glucose
levels
Ovaries Estrogen Stimulates the maturation of Hyposecretion hampers
the female reproductive the ability of a woman
organs and development of to conceive and bear
secondary sex characteristics children
of the female; in cooperation
with progesterone, it causes
menstrual cycle
Progesterone It works with estrogen in Hyposecretion hampers
establishing the menstrual the ability of a woman
cycle to conceive and bear
children
Testes Testosterone Promotes maturation of the In cases of
male reproductive organs, hyposecretion, the man
male secondary sex becomes sterile
characteristics and (sterility)
production of sperm by testes
Pineal gland Melatonin Affects biological rhythms
and reproductive behavior
Thymus gland Thymosin Cause the maturation of T
 lymphocytes

ACID BASE BALANCE

Respiratory Acidosis Compensation: bicarbonate retention

(lungs) Causes: CO2 excess (hypoventilation), COPD, drug overdose Compensated:↑HCO3- + ↓pCO2 + pH
(morphine, barbiturates, opiates) <7.4
Alkalosis Compensation: bicarbonate excretion
Causes: CO2 loss (hyperventilation) Compensated:↓HCO3- + ↓pCO2 + pH
>7.4
Metabolic Acidosis Compensation: hyperventilation
(kidney) Causes: HCO3- deficiency, DKA (normochloremic acidosis), Compensated: :↓HCO3- + ↓pCO2 + pH
renal failure, diarrhea (↓HCO3-) <7.4
Alkalosis Compensation: hypoventilation
Causes: HCO3- excess, vomiting (↓Cl-), hypchloremia, Compensated:↑HCO3- + ↓pCO2 + pH
hypokalemia >7.4

SIGNIFICANT PROTEIN ELECTROPHORETIC PATTERN

FRACTION SPECIFIC PROTEIN

Albumin Albumin
Alpha1 globulin Alpha1 antitrypsin, lipoproteins
Alpha2 globulin Ceruloplasmin, haptoglobin, alpha2 macroglobulin, lipoproteins
Beta globulin Transferrin, Hemopexin, Complement system, lipoproteins
Gamma globulin Immunoglobulins

CHARACTERISTICSOF HUMAN APOLIPOPROTEINS AN THEIR FUNCTION (Bishop 3rd ed.)

APOLIPOPROTEIN FUNCTION MAJOR SOURCE

Apo A-I Major Structural Protein in HDL Liver and Intestines
Activates LCAT
Ligand for HDL binding
Apo A- II Structural Protein in HDL Liver
Activates LCAT
Enhances hepatic triglyceride lipase activity
Apo A- IV Component of intestinal lipoproteins Intestine
Apo B-100 Major structural proetein in VLDL and LDL Liver
Ligand for the LDL receptor
Apo B-48 Primarily structural protein in chylomicrons Intestine
Apo C-I Activates lipoprotein lipase Liver
Apo C-II Activates lipoprotein lipase Liver
Activates LCAT
Apo C – III Inhibits lipoprotein lipase Liver
Inhibits receptor recognition of apo E
Apo E2,3,4 Binds to LDL-receptor and remnant –receptor Liver
Apo (a) Structural protein for Lp (a) Liver
May inhibit plasminogen binding

DRUGS OF ABUSE

Stimulants  Methamphetamine chloride (shabu)

 Methylene dioxymethamphethamine (ecstacy)
 Amphetamine sulfate
 Dextroamphetamine sulfate
 Phenmetrazine
 Methylphenidate
 Coacaine
 Caffeine
  Theophylline
 Theobromine
 Nicotine
Depressants  Barbiturates
 Benzodiazepams (ativan, valium)
 Methaqualone (Quaalude)
 Gamma hydroxybutyrate
Hallucinogen/Psychedelics  Lysergic acid diethylamide
 Mescaline
 Psilocybin
 Phencyclidine
 Marijuana
 Hashish
Narcotics/Opoids/Opiates  Opium
 morphine
 heroin
 codeine
 methadone
 mepiridine (Demarol)
 proxyphene (Darvon)
 Fentanyl
Anesthetics  Ketamines
Other Compounds  Anabolic steroids
 Inhalants

THERAPEUTIC DRUGS

Cardioactive Lidocaine
Procainamide
Quimidine
Propranolol
Amiodarone
Verapamil
Digoxin
Digitoxin
disopyramide

Aniconvulsants/anti-epileptic Phenobarbital (Antrocol)

Used in treatment of seizure disorders, in particular grand Phenytoin (Dilantin)
mal, petit-mal(absence of seizure), psychomotor seizure and Valproic acid ( Depakene)
other generalized seizure disorder such as tic douloreux Carbamazepine ( Tegretol)
(trigeminal neuralgia) Ethosuximide (Zarontin)
Neurotontin (Gabapentin)
Iamotrigine (Lactamical)
Topiramate
Felbamata
Antidepressants / Manic depression treatment Lithium – bipolar disorder (manic depressive psychosis)
Tricyclic Antidepressant (TCA) – amitriptyline, imipramine,
nortriptyline, desipramine,
Doxepin
Immunosuppressive Cyclosporine
Tacrolimus (FK-506)
Rapamycin (Sirolimus)
Mycophenolate mofetil
Leflunamide (LFM)
Prednisone
Cyclophosphanamide (Cytoxan)
Anti-psychotic Phenothiazines (chlorpromazine, thiordazine, fluphenazine)
Butyrophenones (haloperidol)
Risperdal
Olanzapine (Zyprexa)
Ouetiapine (Seroquel)
 Aripirazole (Abilify)
Anti-neoplastic Methotrexate (also an immunosuppressant)
Busulfan
Anti-inflammatory Acetylsalicylic acid
Acetaminophen
Antibiotics Aminoglycosides
Vancomycin
Chloramphenicol
Trimetophrim lactate
Bronchodilators/ Anti-asmathics Theophylline

CHEMICAL TEST

QUALITATIVE TEST FOR PROTEIN

Heller’s
Robert’s White ring at the zone of contact
Spiegler’s
Biuret Violet for albumin
Rose for albuminoses and peptones
Heat and acetic acid
SSA
Purdy’s White turbidity/cloudiness
Potassium ferrocyanide
Picric acid
QUANTITATIVE TESTS FOR PROTEIN
Esbach’s 24◦ - read height of coagulum
Kwilecki’s 72◦C for 5 minutes – read height of coagulum
Tsuchiya’s Same as Esbach’s
Kingsburry-Clark Degree of turbidity is measured by comparison with standard turbidities
Biuret Uses the same principle as that used for serum protein which depends upon the
presence of peptide linkages in protein
SUGARS
Benedict’s Reducing substances
Green-orange-red
Osazone or Phenylhydrazine (Kowarsky) Glucose, fructose, lactose and pentose
Crystalline needles
Nylanders Glucose and other reducing subs
Brown to black color
Moore heller Glucose and other reducing subs
Canary yellow to black
Borchardt’s Fructose
Seliwanoff Red color
Resorcinol-HCL
Rubner’s Lactose: Brick red color w/ red ppt
Glucose: Red color w/ yellow ppt
Bial Orcinol Green sol’n
Tauber’s Cherry red
KETONES
Frommer’s Acetone
Purplish red ring
Rothera’s Acetone and acetoacetic acid
Rose or purple ring
Lange Acetone and acetoacetic acid
Purple ring
Acetest Acetone and acetoacetic acid
Ketostix Purple color
Gerhardt’s Acetoacetic acid
Bordeaux red color
BILE PIGMENTS
(Bilirubin, Urobilinogen, Urobilin)
Gmelin Bile pigments
Play of colors
Smith Bile pigments
Emerald green
Harrison’s spot Bile pigments
Blue to green color
Ictotest Bile pigments
Blue to purple mat
Wallace and Diamond Urobilinogen
Cherry red color
Schlesinger Urobilin
Greenish fluorescence
HEMOGLOBIN
Benzidine Green-blue
Guiac Blue
Ortho-toluidine Blue
MELANIN
Screening test Urine will turn to black
Thormahlen (Fresh Urine) Dark green or blue color
Blackberg and Wanger (24hr urine) Brown to black ppt
CHLORIDE
Fantus Reddish ppt
Mercurimetric titration Blue-violet colored complex
Schales and Schales
CALCIUM
Sulkowitch Precipitation

CELLS AND INCLUSIONS SEEN IN SYNOVIAL FLUID

CELL/INCLUSION DESCRIPTION SIGNIFICANCE

Neutrophil Polymorphonuclear leukocyte Bacterial sepsis
Crystal-induced inflammation
Lymphocyte Mononuclear leukocyte Nonseptic inflammation
Macrophages (monocyte) Large mononuclear leukocyte, may be vacuolated Normal
Viral infections
Synovial lining cell Similar to macrophage, but may be multinucleated, Normal
resembling a mesothelial cell Disruption from arthrocentesis
LE cell Neutrophil containing characteristic ingested “round Lupus erythematosus
body”
Reiter cell Vacuolated macrophage ingested neutrophils Reactive arthritis (infection in another
part of the body)
RA (ragocyte) cell Neutrophil with dark cytoplasmic granules Rheumatoid arthritis
containing immune complexes Immunologic inflammation
Cartilage cells Large, multinucleated cells Osteoarthritis
Rice bodies Macroscopically resemble polished rice Tuberculosis
Microscopically show collagen and fibrin Septic and rheumatoid arthritis
Fat droplerats Retractile intracellular and extracellular globules Traumatic injury
Stains with sudan dyes Chronic inflammation
Hemosiderin Inclusion within clusters of synovial cell Pigmented villonodular synovitis

SIGNIFICANCE OF CHEMICAL TESTING OF PLEURAL FLUID

TEST SIGNIFICANCE
Glucose decreased in rheumatoid inflammation
decreased in purulent infection
Lactate Elevated in bacterial infection
Triglycerides Elevated in chylous effusions
pH Decreased in pneumonia not responding to antibiotics
Markedly decreased with esophageal rupture
ADA Elevated in tuberculosis and malignancy
Amylase Elevated in pancreatitis, esophageal rupture, and malignancy
CONVERSION OF TRADITIONAL UNITS TO SI UNITS FOR COMMON CHEMISTRY ANALYTES

CONVENTIONAL/ SI UNITS CONVERSION FACTORS

TRADITIONAL
Albumin g/100 mL g/dL 10
AST U/L (mu/mL) µkat/L 0.0167
Ammonia µg/dL µmol/L 0.587
Bicarbonate mEq/L mmol/L 1.0
Bilirubin mg/dL µmol/L 17.1
BUN mg/dL mmol/L 0.357
Calcium mg/dL mmol/L 0.25
Chloride mEq/L mmol/L 1.0
Cholesterol mg/dL mmol/L 0.026
Cortisol µg/dL µmol/L 0.0276
Creatinine mg/dL µmol/L 88.4
Crea Clearance mL/min mL/s 0.0167
Folic acid ng/mL nmol/L 2.27
Glucose mg/dL mmol/L 0.0555
Hemoglobin g/dL g/dL 10
Iron mg/dL µmol/L 0.179
Lithium mEq/L µmol/L 1.0
Magnesium mEq/L mmol/L 0.5
Osmolality mOsm/kg mmol/kg 1.0
Phosphorus mg/dL mmol/L 0.323
Potassium mEq/L mmol/L 1.0
Sodium mEq/L mmol/L 1.0
Thyroxine µg/dL nmol/L 12.9
Total protein mg/dL g/dL 10
Triglycerides mg/dL mmol/L 0.0113
Uric acid mg/dL mmol/L 0.0595
Vitamin B12 ng/mL pmol/L 0.0738
PCO2 mm/Hg kPa 0.113
PO2 mm/Hg kPa 0.133

VIRUSES

DNA VIRUSES RNA VIRUSES

Adenoviridae Arenaviridae
Hepadnaviridae Bunyaviridae
Herpesviridae Caliciviridae
Papovaviridae Coronaviridae
Parvoviridae Filoviridae
Poxviridae Flaviviridae
Paramyxoviridae
Picornaviridae
Reoviridae
Retroviridae
Rhabdoviridae
Togaviridae

VITAMINS and their Clinical Significance

Fat Soluble Vitamins Chemical Name function Deficiency

1. Vitamin A Retinol Maintenance of good vision, Night blindness
resistance infection Growth retardation
 Abnormal taste response
Dermatitis
2. Vitamin E Tocopherols (α,β,γ,δ) Antioxidant; for cellular Mild hemolytic anemia
respiration (newborn), RBC fragility,
ataxia
3. Vitamin D2 Ergocalciferol, Rickets (young),
Cholecalciferol osteomalacia (adult)
4. Vitamin D3 Activated sterol – 1, 25 Absorption of dietary Hypocalcemia
dihydroxycholecalciferol calcium
5. Vitamin K Phytomenadione Cofactor of procoagulants Bleeding disorder,
like prothrombin hemorrhage
Water Soluble Vitamins
1. Vitamin B1 Thiamine Enzyme cofactor Infants: Dyspnea, cyanosis,
diarrhea and vomiting
Adults: beri-beri, Wernicke-
Korsakoff syndrome
(apathy, ataxia and visual
problems
2. Vitamin B2 Riboflavin Enzyme cofactor Angular stomatitis,
dermatitis, photophobia
3. Vitamin B3 Pantothenic acid Enzyme cofactor Depressed immune system,
muscle weakness
4. Vitamin B6 Pyridoxine, Pyridoxal Enzyme Cofactor Infants: irritability, seizures,
anemia
Adults: facial seborrhea
5. Vitamin B12 Cyanocobalamin Syntesis of DNA and Folate Megaloblastic anemia,
neurologic abnormalities
6. Vitamin C Ascorbic acid Hydroxylation of collagen;
participates in redox
reactions
7. Biotin Enzyme cofactor Dermatitis, hair loss,
depression
8. Cartinine Fat Catabolism Muscle weakness, fatigue
9. Folic Acid Pteroylglutamic acid Synthesis of amino acids Megaloblastic anemia
and DNA
10. Niacin/Niacinamide Nicotinic acid/ nicotinamide Enzyme cofactor Pellagra (dermatitis,
disorientation, weight loss)

SURFACE MARKERS ON T AND B CELLS

ANTIGEN CELL TYPE FUNCTION

CD2 Thymocyte, T cells, NK cells Involved in T cell activation
CD3 Thymocyte, T cells Associated with T cell antigen receptor; role in TCR
transduction
CD4 Helper T cells monocyte and macrophages Co-receptor for MHC class II, receptor for HIV
CD5 Mature T cells, thymocytes, subset for B cells Positive or negative modulation of T and B cell receptor
signaling
CD8 Thymocytes subsets, cytotoxic T cells Co-receptor for MHC class I
CD10 B and T cell precursor, bone marrow stromal Protease; marker for pre-B CALLA
cells
CD16 Macrophages, NK cell, neutrophils Low affinity Fc receptor, mediates phagocytosis and
ADCC
CD19 B cells, follicular dendritic Part of B cell co-receptor, signal transduction molecule that
regulates B cell development and activation
CD21 B cells, follicular dendritic, subset of immature Receptor for complement component C3d part of B cell co-
thymocytes receptor with CD19
CD23 B cells, monocytes, follicular dendritic cells Regulation of IgE synthesis; triggers release of GM-CSF
from monocytes
CD25 Activated T, B cells, monocytes Receptor for IL-2
CD44 Most leukocytes Adhesion molecule mediating homing to peripheral
 lymphoid organs
CD45R Different forms on all hematopoietic cells Essential in T and B cell antigen-stimulated activation
CD56 NK cells, subsets of T cells Not known
CD94 NK cells, subsets of T cells Subunit of NKG2-A complex involved in inhibition of
NK cells cytotoxicity
*Cluster of differentiation

DISORDER ANTIBODIES
Myasthenia gravis Acetylcholine receptor blocking antibody
Multiple sclerosis Anti-myelin antibody
Pernicious anemia Anti-intrinsic factor antibody
Anti-parietal cell antibody
Goodpasture’s syndrome Anti-glomerular basement membrane antibody
Primary biliary cirrhosis Anti-mitochondrial antibody
Chronic active hepatitis Anti-smooth muscle antibody
Hashimoto’s thyroiditis Anti-microsomal antibody
Anti-thyroglobulin anibody
Grave’s disease Anti-TSH receptor

MICROSCOPY TECHNIQUES

TECHNIQUE FUNCTION
Bright-field microscopy Used for routine urinalysis
Phase-contrast microscopy Enhances visualization of elements with low refractive indices, such as hyalaine
casts, mucous threads and Trichomonas
Polarizing microscopy Aids in identification of cholesterol in oval fat bodies, fatty casts, and crystals
Dark-field microscopy Aids in identification of Treponema pallidum
Fluorescence microscopy Allows visualization of naturally fluorescent microorganisms or those stained by
fluorescent dye
Interference-contrast Produce a three-dimensional microscopy-image and layer –by-layer imaging of a
sprecimen

ISBT TERMINOLOGY FOR RED BLOOD CELL SURFACE ANTIGENS IN BLOOD GROUP SYSTEM

ISBT SYSTEM NUMBER SYSTEM CHROMOSOMAL NUMBER

001 ABO 9
002 MNS 4
003 P 22
004 Rh 1
005 Lutheran 19
006 Kell 7
007 Lewis 19
008 Duffy 1
009 Kidd 18
010 Diego 17
011 Cartwright 7
013 Scianna 1
014 Dombrock Not known
015 Colton 7
016 Landsteiner-Weiner 19
017 Chido/Roger 6
018 H 19
019 Kx X
020 Gerbich 2
021 Cromer 1
022 Knops 1
023 Indian 11
HOST DEFENSES IN INNATE IMMUNITY

Cells Antigen presenting cells, basophils, eosinophils, mast cells, natural killer cells, phagocytes
Humoral Factor Complement proteins, lactoferrin, lysozyme, pepsin, stomach acidity
Anatomical Barriers Cilia, mucous, skin
Resident flora Mainly non-pathogenic bacteria

HOST DEFENSES IN ADAPTIVE IMMUNITY

Cells B cells, plasma cells, T cytotoxic cells, T helper cells

Humoral Factors Antibodies, cytokines

SELECTIVE MEDIUM FOR N. gonorrhea

THAYER MARTIN MODIFIED THAYER MARTIN LEWIS NEW YORK CITY

MARTIN
Vancomycin inhibit gram Vancomycin + colistin + Vancomycin + colistin + Vancomycin+colistin+trimethroprim
positive nystatin + trimethroprim trimethroprim lactate + lactate + amphotericin B= inhibition
lactate (prevents swarming anisomycin= inhibition of of fungi
of proteins) fungi
Colistin Inhibit gram
negative except N.
gonorrhea
Nystin inhibit fungi

CHANGES IN UNPRESERVED SPECIMEN

ANALYTE CHANGE CAUSE OF CHANGE

Color Modified/darkened Oxidation or reduction of metabolites
Clarity Decreased Bacterial growth and precipitation of amorphous material
Odor Increased Bacterial multiplication/breakdown of urea into NH3
pH Increased Breakdown urea into ammonia by urease-producing bacteria/
loss of CO2
Glucose Decreased Glycolysis and bacterial use
Ketones Decreased Volatilization and bacterial metabolism
Bilirubin Decreased Exposure to light / photo oxidation to biliverdin
Urobilinogen Decreased Oxidation to urobilin
Nitrite Increased Multiplication of nitrate-reducing bacteria
RBCs, WBCs and Casts Decreased Disintegration in dilute and alkaline urine
Bacteria Increased Multiplication

LABORATORY DIFFERENTIATION OF TRANSUDATES AND EXUDATES

TRANSUDATES EXUDATES
Appearance Clear Cloudy
Fluid:serum protein ratio <0.5 >0.5
Fluid:serum LD ratio <0.6 >0.6
WBC count <1000/uL >1000/uL
Spontaneous clotting No Possible
Pleural cholesterol <45-60mg/dL 46-60 mg/dL>
Pleural fluid:serum cholesterol ratio <0.3 >0.3
Plerural fluid:bilirubin ratio <0.6 >0.6
Serum-ascites albumin gradient >1.1 <1.1
COLOR REACTION FOR QUANTITATION OF BILIRUBIN

Bilirubin + diazotized sulfanilic acid azobilirubin

 Color is proportional to the concentration of bilirubin

ASSAY EVELYN-MALLOY JENDRASSIK-GROF

pH Acid Alkaline
Dissociating agent Methanol Caffeine-sodium benzoate
Diazo product Red or reddish-purple color (absorption Blue (maximum absorbance around 600
maximum in the region of 560 nm) nm)

MAJOR HISTOCOMPATIBILITY COMPLEX

CLASS I CLASS II CLASS III

Genetic loci HLA – HLA- C2
A,B,C DP,DQ,DR C4
Chain struncure Light chain Light chain FACTOR B
B2 microglobulin B chain TNF
Cell distribution ALL nucleated cell B cell and APC
Present antigen CD8 of Tc cells CD4 of Th cells

NOBEL PRIZE WINNERS IN IMMUNOLOGY (Stevens)

YEAR SCIENTIST RESEARCH

1901 Emil Von Behring Serum antitoxins
1905 Robert Koch Cellular immunity in TB
1908 Elie Metchnikoff, Paul ehrlich Phagocytosis
Immunity
1913 Charles Richet Anaphylaxis
1919 Jules Bordet Complement
1930 Karl Lansteiner Human blood group antigens
1960 Macfarlane Burnet, Peter Medawar Discovery of immunologic tolerance
1972 Gerald Edelman, Rodney Porter Structure of antibodies
1977 Rosalyn Yalow Radioimmunoassay
1980 George Snell, Jean Dausset, Baruj Major Histocompatibility complex
Beneceraf
1984 Niels Jerne Immunoregulation
Georges Kohler, Cesar Milstein Monoclonal antibody
1987 Susumu Tonegawa Antibody Diversity
1991 Edward Donnall Thomas, Joseph Transplantation
Murray
1996 Peter Doherty, Rolf Zinkernagel Cytotoxic T cell Recognition of variety of infected cells
2008 Francoise Barre-Sinoussi, Luc Human immunodeficiency virus
Montagnier

HISTORICAL PERSPECTIVE (Stanley)

1798 Edward Jenner, an English countryside physician demonstrated that protection from cowpox could be generated
by the transfer of postural material from a cowpox lesion instead of the more hazardous smallpox lesion

1880 Louis Pasteur demonstrated that injection of killed microbes provided protection upon subsequent exposure to
live counterpart
1888 Elie Metchnikoff demonstrated that certain blood cells ingest foreign material
1894 Jules Bordet discovered complement
1897 Robert Kaus discovered precipitins
1901 Emil von Behring had the distinction of being awarded as the first immunology-related Nobel Prize for his
works on serum therapy
1984 Discovery of the T cell receptor gene
1987 Susumu Tonegawa was awarded the Nobel Prize for his 1978 discovery of the genetic principles underlying the
generation of antibodies with different specificities

FIRE/EXPLOSIVE HAZARD

JCAHO (Joint Commission on Accreditation of Healthcare Organization) requires ALL healthcare institution post
evacuation routes and detailed plans to follow in case of a fire

WHEN FIRE IS DISCOVERED: USE OF FIRE EXTINGUISHER

R – rescue P – pull pin
A – alarm/activate A- aim the base of the fire
C – contain/close S – squeeze handles
E – extinguish S – sweep nozzle side to side

TYPES OF FIRE AND FIRE EXTINGUISHER

FIRE EXTINGUISHING MATERIAL TYPES OF FIRE EXTINGUISHER

TYPE
Class A Wood, paper, clothing Class A Water
Class B Flammable organic chemicals Class B Dry chemicals, CO2,
foam/halon
Class C Electrical Class C Dry chemicals, CO2, halon
Class D Combustible materials NONE Sand/dry powder
Class ABC Dry chemicals

POTASSIUM
HYPOKALEMIA HYPERKALEMIA
 Intracellular shift  True Hyperkalemia
 Alkalosis, periodic paralysis, Beta-2-  Due to extracelluar shift
agonists, Barium poisoning, Insulin - Acute acidosis (especially inorganic
overdose, Nutritional recovery state acidosis)
 Poor intake - Catabolic states, periodic paralysis,
 Vomiting, Diarrhea, intestinal drainage/ succinylcholine, muscle or cellular
tumor, gastric suction,laxative abuse, injury
malabsorption, cancer thereapy – - Chemotheraphy, leukemia
chemotherapy, radiation therapy - Cationic amino acids
 Excessive renal loss - Exercise while using a beta-blocker
 Primary aldosteronism (adrenal adenoma or - Digitalis intoxication
hyperplasia); plasma renin activity is  Due to excessive ingestion: rare if renal
suppressed excretion of K+ is normal; oral or IV
 Secondary aldosteronism (the increase in potassium replacement therapy
aldosterone is secondary to increase in  Decreased renal excretion:
renin) - Hypoaldosteronism: Addison’s disease;
 Malignant Hypertension, renal artery selective hypoaldosteronism
stenosis, reninoma (hyporeninemic hypoaldosteronism,
 Diuretics heparin, congenital adrenal enzyme
 Nephritis deficiencies, angiotensin-converting
 Bartter’s syndrome, Gitelman’s syndrome enzyme inhibitors)
 Excess mineralocorticoids other than - Tubular unresponsiveness to
aldosterone, e.g. Cushing’s syndrome, aldosterone (pseudohypoaldosteronism
ACTH-producing tumor, licorice type I and type II): congenital, salt-
  Chronic meteabolic acidosis, renal tubular losing nephropathy
acidosis -
Acute or chronic renal failure
 Delivery of poorly reabsorbed anions to the -
Potassium-sparing diuretics
diatal tubule, e.g. bicarbonate ketone, -
Antirejection mdeications: ciclosporin,
anions, carbenicillin tacrolimus
 Miscellaneous causes: Magnesium - Severe dehydration
deficiency, acute leukemia, Liddle’s  Pseudohyperkalemia
syndrome  Thrombocytosis, severe leukocytosis, use of
tourniquet with fist exercise, in vitro
hemolysis

TYPES OF CENTRIFUGE

Horizontal/swinging bucket centrifuge Allow the tubes to attain a horizontal position in the centrifuge when spinning and
a vertical position when the head is not moving.
 The specimen cups in the horizontal centrifuge heads are in a vertical
postion when the centrifuge is at rest. During centrifugation, the cups
move to a horizontal position. As the specimen is centrifuged, the particles
being sedimented travel down through the liquid to the bottom of the tube.
When the centrifuge stops and the tubes swing to a vertical position there
may be remixing of the sediment with the supernatant liquid. These
centrifuges are capable of speeds up to about 3000 RPM. Higher speeds
that this will generally cause excessive heat buildup as a result of air
friction.
Fixed-angle/angle head centrifuge Have angled compartments for the tubes and allow small particles to sediment
more rapidly.
 Angle centrifuge heads are capable of higher speed and contain drilled
holes that hold the tubes at a fixed angle (approximately 52 degree angle
with the center shaft around which they rotate). There is much less heat
developed during centrifugation because of very low air friction. During
centrifugation, the particles travel across the column of the liquid to the
side of the tube where they clump together and then rapidly move to the
bottom of the tube.
Ultracentrifugation High-speed centrifuges used to separate layers of different specific gravities. They
are commonly used to separate lipoproteins. The chamber is usually refrigerated to
counter heat produced through friction.

LABORATORY FINDING IN JOINT DISORDER

INFLAMMATORY
NONINFLAMMATORY Immunologic origin Crystal-induced SEPTIC HEMORRHAGIC
Clear, yellow fluid Cloudy, yellow fluid Cloudy or milky fluid Cloudy, yellow-green Cloudy, red fluid
fluid
Good viscosity Poor viscosity Low viscosity Variable viscosity Low viscosity
WBC <1000 uL WBC 2000-75,000 uL WBC up to 100,000 WBC 50,000-100,000 WBC equal to blood
uL uL
Neutrophil <30% Neutrophil >50% Neutrophil <70% Neutrophil >75% Neutrophil equal to
blood
Normal glucose Decreased glucose Decreased glucose Decreased glucose Normal glucose
(similar to blood glucose) level level level (similar to blood
Possible Crystal present Positive culture and glucose)
autoantibodies gram stain

COMMON FECAL TEST FOR DIARRHEA

SECRETORY OSMOTIC
 Stool cultures  Microscopic fecal fat
 Ova and parasite examination  Muscle fiber detection
  Rotavirus immunoassay  Qualitative and Qualitative fecal fats
 Fecal leukocyte  Trypsin screening
 Clinitest
 D-xylose tolerance test
 Lactose tolerance test
 Fecal electrolytes
 Stool pH
 Fecal osmolality

SIMULATION NO.2

FILARIAL HABITAT VECTOR SPECIMEN MICROFILARIA PERIODICITY

WORM
Wuchereria Lymphatics Aedes, Anopheles Blood Sheated, nuclei Nocturnal
brancrofti absent in tail
Brugia malayi Lymphatics Mansonia sp. Blood Sheated, tail Subperiodic
With 2 separate Nocturnal
Nuclei
Loa loa Subcutaneous Chrysops sp. Blood Sheated, nuceli Diurnal
tissue Tabanid or Continuous to
Mango fly The tip of the tail
Onchocerca Subcutaneous Simulium sp. Skin snips Unsheated, Nonperiodic
volvulus tissue blackfly Nuclei absent in
Tail

SODIUM
HYPONATREMIA HYPERNATREMIA
 Due to Na+ loss  Reduced water intake
 Thiazide diuretics in the presence of ADH  Defective thirst due to altered mental state
 Saline infusion in the presence of ADH or thirst center defect
 Due to water retention  Inability to drink water
 Excessive water intake: primary polydipsia  Lack of access to water
 Advance renal failure  Increase water loss (water intake must be impaired)
 Appropriate ADH secretion: edema-forming  Gastrointestinal loss: vomiting, osmostic
states (CHF, nephritic syndrome, ascites). diarrhea
Salt depletion states (GI loss, diuretic  Cutaneous loss: sweating and fever
therapy, aldosterone deficiency,  Respiratory loss: hyperventilation and fever
hypothyroidism)  Renal loss: diabetes insipidus, osmotic
 Inappropriate ADH secretion diuresis
 Tumors: cancers of the lung, pancreas,  Increased sodium content of the body (water intake
duodenum, ureter, bladder, prostrate, must be impaired)
lymphoma, thymoma, mesothelioma,  Increased intake
Ewing’s Sarcoma  Hypertonic saline or sodium bicarbonate
 Intrathoracic causes: bacterial and viral infusion
pneumonia, tuberculosis, lung abscess,  Ingestion of sea water
aspergillosis, asthma, positive pressure  Renal salt retention; usually in response to
breathing, pneumothorax, cystic fibrosis primary water deficit
CNS abnormalities: encephalitis meningitis,
brain tumors and abscess, head trauma,
subdural hematoma, cerebrovascular
accidents, Guillen-Barré syndrome, acute
intermittent porphyria, brain atrophy,
schizophrenia, hydrocephalus, acurte
psychosis, multiple scheloris, cavernous vein
thrombosis, lupus cerebritis
 Shy-Drager syndrome, Rocky Mountain
 spotted fever, delirium tremens, seizure
disorder
 Drugs: arginine vasopressin and its analogs,
sulfonylureas, tricyclic antidepressants,
clofribate, carbamazepine, vinca alkaloids,
cyclophosphamide, selective serotonine
reuptake inhibitors, opiates, phenothiazines,
haloperidol
 Surgical and emotional stress, emesis
 Endocrine causes: glucocorticoid deficiency
and myxedema

HYPONATREMIA with Normal Renal Function

CAUSE SERUM Na Urine Na 24-hour Na Urine osmolality Serum K

1.Overhydration Low Low Low Low Normal or Low
2.Diuretics Low Low High Low Low
3.SIADH Low High High High Normal or Low
4.Adrenal Failure Mildly Elevated Normal High High
5.Bartter’s Low Low High Low Low
syndrome
6.Diabetic low Normal Normal Normal High
hyperosmolality

CHARACTERISTICS OF HUMAN PLASMODIUM SPP.

CHARACTERISTICS P.vivax P. falciparum P. malariae P. ovale

Persistence of YES NO NO YES
exoerythrocytic cycle
Length of replication 44-48 36-48 72 48
cycle (hours)
Schuffner’s dots Present in all infected Absent Absent Usually present in all
RBC except early ring infected RBCs
forms
Number of merozoites 16 (12-24) Schizonts not seen in 8 (6-12) 8 (8-12)
in mature schizonts peripheral blood
Important criteria for Infected RBCs Multiple ring forms Infected RBCs normal Infected RBCs
identification enlarged, trophozoites seen in a single RBC, size and color, enlarged, often oval
irregularly shaped, crescent shaped trophozoites compact shaped with fimbriated
Schuffner’s dots gametocytes, and ring and band forms may be edges, trophozoites
shaped young seen irregular shaped
trophozoites are only Schuffner’s dots
forms seen

CHARACTERISTICS OF ACUTE-PHASE REACTANTS

PROTEIN RESPONSE NORMAL INCREASE FUNCTION

TIME CONCENTRATION
(hr) (mg/dL)
C-reactive protein 6-10 0.5 1000x Opsonization, activation
Serum amyloid A 24 3.0 1000x Removal of cholesterol
Alpha1-antitrypsin 24 200-400 2-5x Protease inhibitor
Fibrinogen 24 110-400 2-5x Clot formation
Haptoglobin 24 40-200 2-10x Binds hemoglobin
Ceruloplasmin 24 20-40 2x Binds copper and oxidizes iron
Complement C3 48-72 60-140 2x Opsonization lysis
Mannose binding 48-72 0.15-1.0 ? Complement activation
protein

ABO DISCREPANCIES

GROUP 1 DISCREPANCIES: WEAKLY REACTING OR MISSING ANTIBODIES

1. Newborn
2. Elderly patient
3. Patient with leukemia demonstrating hypogammaglobulinemia
4. Patient with lymphomas demonstrating hypogammaglobulinemia
5. Patients using immunosuppressive drugs that yield hypogammaglobulinemia
6. Patients with congenital agammaglobulinemia
7. Patients with immunodeficiency diseases
8. Patients with bone marrow transplantation (develop hypogammaglobulinemia)

GROUP II DISCREPANCIES: WEAKLY REACTING OR MISSING ANTIGENS

1. Subgroup of A and or B
2. Leukemias (weakened A or B antigen)
3. Hodgkin’s disease
4. Excess amount of blood group-specific soluble subsntances (BGSS) present in the plasma in association with
certain diseases such as carcinoma of the stomach and pancreas
5. Acquired B phenomenon
6. Antibodies to low incidence antigens

GROUP III DISCREPANCIES: PROTEIN OR PLASMA ABNORMALITIES RESULTING TO ROULEAUX

FORMATION

1. Elevated levels of globulin from certain disease states : MM, waldenstrom’s macroglobulinemia, other plasma
cell dyscarias, advanced hodgkin’s lymphoma
2. Elevated levels of fibrinogen
3. Plasma expanders such as dextran and polyvinylpyrrolidone (PVP)
4. Wharton’s jelly

GROUP IV DISCREPANCIES: MISCELLANEOUS

1. Polyagglutination
2. Cold reactive antibodies
3. Unexpected ABO isoagglutinins
4. Antibodies other than anti-A and anti-B may react to form antigen-antibody complexes that may then absorb into
patient red cells
5. RBCs with the cis AB phenotype.

CHARACTERISTICS OF SELECTED PLASMA PROTEINS (Bishop 6th ed.)

Prealbumin (transthyretin) Indicator of malnutrition: binds thyroid hormone and retinol-binding protein.
Albumin
 α1-antitrypsin Acute-phase reactant; protease inhibitor
 α1- fetoprotein Principal fetal protein
 α1- acid glycoprotein Acute-phase reactant
 (orosomucoid)
 α1- lipoprotein Transposrts lipid
(HDL)
 α1-antichymotrypsin Inhibits serine proteinases (ie, chymotrypsin)
 Inter-α-trypsin Inhibits proteinases (ie, trypsin)
inhibitor
 Gc-globulin Binds vitamin D and actin
α2-Globulins
 Haptoglobins Acute-phase reactant; binds hemoglobin
 Ceruloplasmin Peroxidase activity; contains copper
 α2- Macroglobulins Inhibits thrombin, trypsin, pepsin
β-Globulins
 Pre-β-Lipoproteins Transports lipids (primarily triglycerides)
(VLDL)
 Transferrin Transposrts Iron
(Sideroplasmin)
 Hemopexin Binds Heme
 β-Lipoproteins(LDL) Transposrts lipid (primarily cholesterol)
 β2- Component of human leukocyte antigen (HLA) molecules class 1
Microglobulin(B2M)
 Complement Immune response
 Fibrinogen Precursor of Fibrin clot
 C-reactive Protein Acute-phase reactant; motivates phagocytosis in inflammatory disease (Bishop) (Henry-γ)
(CRP)
γ-Globulins
 Immunogloblulin G Antibodies
 Immunoglobulin A Antibodies (in secretions)
 Immunoglobulin M Antibodies (early reponse)
 Immunoglobulin D Antibodies
 Immunoglobulin E Antibodies (allergy)

NCEP GUIDELINES FOR ACCEPTABLE MEASUREMENT OF ERROR (Henry)

ANALYTE TOTAL ERROR BIAS CV

Cholesterol < 9% <3% <3%
Triglycerides <15% <5% <5%
HDL-cholesterol <13% <5% <4%
LDL-cholesterol ≤12% ≤4% ≤4%
┼

ENZYME CLASSES

CLASS CATEGORY TYPE OF REACTION CATALYZED EXAMPLES

1 Oxireductase Oxidation/reduction reduction Lactate dehydrogenase
Glucose-6-phosphate dehydrogenase
Glutamate dehydrogenase
2 Transferase Transfer of Intact group of atoms from one Lactate dehydrogenase
molecule to another Glucose-6-phosphate
Dehydrogenase
Glutamate dehydrogenase
3 Hydrolase Cleavage of bonds with water Alkaline phosphatase
Acid phosphatase
Amylase
Triacylglycerol lipase
Cholineesterase
Chymotypsin
Elastase-1
5-nucleotidase
Trypsin
 4 Lyases Cleavage of C-C, C-O, C-N or other types of Aldolase
bonds; does not involve water
5 Isomerase Convert one isomer to another Triosephosphate isomerase
6 Ligases Bond formation between two groups of atoms; Glutathione
with ATP as energy source

BACTERIAL IDENTIFICATION AND STRATEGIES

TEST PRINCIPLE POSITIVE NEGATIVE POSITIVE NEGATIVE

RESULT RESULT CONTROL CONTROL
Acetamide This test is used to determine Deamination of No color change Pseudomonas Stenotrophomonas
Utilization the ability of an organism to the acetamide aeruginosa maltophilia
use acetamide as the sole resulting in the
source if carbon. Bacteria that blue color
can grow on this medium
deaminate acetamide to release
ammonia. The production of
ammonia results in the a pH-
driven color change of the
medium from green to royal
blue
Actetate The test is used to determine if Medium No growth or Escherichia coli Shigella flexneri
Utilization an organism can use acetate as becomes growth with no
the sole source of carbon. If alkaline(blue) indicator change
so, breakdown of sodium because of the to blue
acetate causes the pH of the growth of the
medium to shift toward the organism
alkaline range, turning the
indicator from green to blue
Bacitracin The test is used to determine Any zone of No zone of Streptoccocus Streptococcus
Test the effect of a small amount of inhibition inhibition pyongenes agalactiae
bacitracin (0.04 U) on an around the disk
organism. Streptococcus
pyogenes is inhibited by the
small amount of bacitracin in
the disk other beta-hemolytic
streptococci usually are not
Bile Esculin Gram-positive bacteria other Blackening of No blackening Enterococcus Streptococcus
Agar than group D streptococci and nthe agar slant of the medium faecalis mitis
enterococci are inhibited by
the bile in this medium. Group
D streptococci and enterococci
can grow in the presence of
40% bile and hydrolyze
esculin, they subsequently turn
the indicator, ferric ammonium
citrate, a dark brown color.
This dark brown color results
from the combination of
esculetin (end product of
esculin hydrolysis) and ferric
ions to form a phenolic iron
complex
Bile Solubility The solubility test Colony Intact colonies Streptococcus Enterococcus
Test differentiates Streptococcus disintegrates; an pneumoniae faecalis
pneumonia (positive) from imprint of the
alpha-hemolytic streptococci lysed colony
(negative). Pneumococcai may remain
colonies are rapidly lysed by within the zone
bile o a solution of a bile salt
such as sodium desoxylate.
Lysis depends on the presence
 of an intracellular autolytic
enzyme. Bile salts lower the
surface tension between the
bacterial cell membrane and
the medium, thus accelerating
the organism’s natural
autolytic process.
Butyrate Disk The butyrate disk is a rapid Development of No color change Moraxella Neisseria
test for the detection of the a blue color catarrhalis gonorrhea
enzyme butyrate esterase in during 5-minute
identifying Moraxella incubation
(Branhamella) catarrhalis. If period.
bromo-chloro-indolbutyrate
impregnated in the disk is
hydrolyzed by the enzyme, a
blue-colored indigo compound
is formed.
CAMP Certain organism (including Enhanced No enhancement Streptococcus Streptococcus
group B streptococci) produce hemolysis is of hemolysis agalactiae pyogenes
diffusible extracellular protein indicated by an
(CAMP factor) that acts arrowhead
synergistically with the beta- shaped zone of
lysin of S.aureus to cause beta hemolysis
enhanced lysis of the red blood at the juncture
cells. of the two
organism
Catalase Test The enzyme catalase mediates Copious bubbles No or few Staphylococcus Steptococcus
the breakdown of hydrogen produced bubbles produce aureus pyogenes
peroxide into oxygen and
water. The presence of the
enzyme in a bacterial isolate is
evident when a small inoculum
is introduced into hydrogen
peroxide, and the rapid
elaboration of oxygen bubbles
occurs. The lack of catalase is
evident by a lack of or weak
bubble formation
Cetrimide This test is used to determine Growth No growth Pseudomonas Escherichia coli
the ability of an organism to aeruginosa
grown in the presence of
cetrimide, a toxic substance
that inhibits the growth of
many bacteria
Citrate This test is used to determine Growth in the Absence of Klebsiella Escherichia coli
Utilization the ability of an organism to medium with or growth pneumoniae
utilize sodium citrate a sits without a
only carbon source and change in the
inorganic ammonium salts as color of the
its only nitrogen source. indicator. The
Bacteria that can grow on this color change of
medium turn the bromthymol the indicator is
blue indicatoe from green to due to acid or
blue. alkali
production of
the test
organism as it
grows on the
medium.
Growth usually
results in the
bromthymol
blue indicator
turning from
green to blue.
Coagulase The test is used to differentiate Clot of any size No clot Staphylococcus Staphylococcus
Test Staphylococcus aureus aureus epidermidis
(positive) from coagulase
negative staphylococci
(negative). S.aureus produces
two forms of coagulase:
boucnd and free. Bound
coagulase or clumping factor
is bound to the bacterial wall
and reacts directly with
fibrinogen. This results in an
alteration of fibrinogen so that
it precipitates on the
staphylococcal cells, causing
the cells to clump when a
bacterial suspension is mixed
with plasma. The presence of
bound coagulase correlates
well with free coagulase, an
extracellular protein enzyme
that causes the formation of
clot when S.aureus colonies
are incubated with the plasma.
The clotting mechanism
involves activation of a plasma
coagulase-reacting factor
(CRF), which is modified or
derived thrombin molecule, to
form a coagulase-CRF
complex. This complex in turn
reacts with fibrinogen to
produce fibrin clot.
Decarboxylase This test measures the Alkaline No color change Lysine: Lysine:
Test enzymatic ability of an (purple) color or acid (yellow) Klebsiella Enterobacter
(Moeller’s organism to decarboxylate (or change color in testand pneumoniae cloacae
Method) hydrolyze) an amino acid to compared with control tube Ornithine: Ornithine:
form an amine. the control tube. Enterobacter Klebsiella
Decarboxylation, or cloacae pneumoniae
hydrolysis, of the amino acids Arginine: Arginine:
results in an alkalin pH Enterobacter Klebsiella
change. cloacae pneumoniae
Base: - Base: Klebsiella
pneumoniae
DNA This test is used to determine When DNA is If there is no Staphylococcus Staphylococcus
Hydrolysis the ability of an organism to hydrolyzed, degradation of aureus. epidermidis
hydrolyze DNA. The medium methyl green is DNA, the
is pale green because of the released and medium remains
DNA-methyl green complex. combines with green.
If the organism growing on the highly
medium hydrolyses DNA, the polymerized
green color fades and the DNA at a pH of
colony is surrounded by a 7.5, turning the
colorless zone. medium
colorless around
the test
organism.
Esculin The test is used to determine Blackened No blackening Klebseilla Shigella flexneri
Hydrolysis whether an organism is able to medium which and no loss of pneumoniae
hydrolyze the glycoside would also show fluorescence
esculin. a loss of under the
fluorescence Wood’s lamp,
under the or slight
Wood’s lamp blackening with
no loss of
fluorescence
 under wood’s
lamp
Fermentation These media are used to Indicator Growth but no Dextrose: Pseudomonas
Media: determine the ability of an changes to pink change in color. positive, with aeruginosa
Peptone organism to ferment a specific with or without Medium gas: Escherichia
medium with carbohydrate that is gas formation in remains clear to coli
Andrade’s incorporated in a basal Durham tube straw-colored. Positive, no gas:
indicator (for medium, thereby producing Shigella flexneri
enteric and acid with or without visible
coryneforms) gas.
Fermentation These media are used to Indicator change Growth but no Sorbitol: Streptococcus
Media: Heart determine the ability of an to yellow change in color. Streptococcus mitis
infusion organism to ferment a specific Medium mutans
broth with carbohydrate that is remains purple.
bromcresol incorporated in a basal
purple medium, thereby producing
indicator (for acid with or without visible
streptococci gas.
and
enterococci)
Flagella Stain A wet-mount technique for EXPECTED RESULTS: Peritrichous: Klebsiella
(Wet Mount staining bacterial flagella is Observe the slide and note the Escherichia coli pneumonia
Technique) simple and is useful when the following: Polar:
number and arrangement of  Presence or absence of Pseudomonas
flagella are critical in flagella aeruginosa
indentifying species of motile  Number of flagella per cell
bacteria.  Location of flagella per
cell
Growth at 42◦ The test is used to determine Good growth No Growth Pseudomonas Pseudomonas
C the ability of an organism tog aeruginosa flourescens
row at 42ᵒC
Gelatin The test is used to determine Partial or total Complete Proteus vulgaris Enterobacter
Hydrolysis the ability of an organism to liquefaction of solidification of aerogenes
produce the proteolytic the inoculated tube at 4ᵒC
enzymes (gelatinases) that tube (the control
liquefy gelatin. must be
completely
solidified at 4ᵒC
within 14 days)
Hippurate The end products of hydrolysis Deep purple Colorless or Streptococcus Streptococcus
Test of hippuric acid include color slightly yellow agalactiae pyogenes
glycine and bezoic acid. pink color
Glycine is deaminated by the
oxidizing agent nihydrin,
which is reduced during the
process. The end products of
the ninhydrin oxidation might
react with any free amino
acids present in growth media
or other broths.
Indole The test is used to determine Pink-to-wine No color change Kovac’s Kovac’s method:
Production the ability of an organism to colored ring after the method: Klebsiella
split tryptophan to form the after the addition of Escherichia coli pneumonia
compound indole addition of appropriate Ehrlich’s Ehrlich’s method:
appropriate reagent method: CDC group EO-2
reagent. Elizabeth kingie
meningoseptica
LAP Test The test is used to identify Development of No color change Enterococcus Leuconostoc sp.
catalase-negative gram a red color or development faecalis
positive cocci. The Lap disk is within 1 minute of a slight
a rapid test for the detection of after adding yellow color
the enzyme leucine cinnamaldehyde
aminopeptidase. Following reagent
hydrolysis of the substrate by
 the enzyme, the resulting β-
naphthylamine produces a red
color upon the addition of
cinnamaldehyde reagent
Litmus Milk This test is used to determine EXPECTED RESULTS
an organism’s ability to Appeareance of Indicator ( Litmus Dye )
metabolize litmus milk.
Fermentation of lactose is COLOR Ph CHANGE TO RECORD
evidenced by the litmus milk Pink, mauve Acid Acid (A)
turning pink as a result of acid Blue Alkaline Alkaline (K)
production. If sufficient acid is Purple (identical to No change No change
produced, casein in the milk is uninoculated control)
coagulated, solidifying the White Independent of pH, Decolorized
milk. With some organisms, result of reduction of
the curd shrinks and whey is indicator
formed at the surface. Some
bacteria hydrolyze casein,
causing the milk to become Appearance of Milk
straw-colored and resemble Consistency of MILK OCCURS WHEN pH
turbid serum. Additionally, is
some organism reduce litmus, Coagulation or clot Acid or alkaline Clot
in which case the medium Dissolution of clot Acid Digestion
becomes colorless in the with clear, grayish,
bottom of the tube watery fluid and a
shrunken, insoluble
pink clot
Dissolution of clot alkaline Peptonization
with clear, grayish,
watery fluid and a
shrunken, insoluble
blue clot

QUALITY CONTROL:
ALKALINE: Alcaligenes faecalis
ACID: Enterococcus faecium
PEPTONIZATION: Burkholderia cepacia
The microdase test is a rapid Development of No color change Micrococcus Staphylococcus
method to differentiate blue to purple- luteus aureus
Staphylococcus from blue color
Micrococcus by detection of
the enzyme oxidase. In the
presence of atmospheric
oxygen, the oxidase enzyme
reacts with the oxidase reagent
and cytochrome C to form the
colored compound,
indophenols.
Motility These tests are used to In true motility, In Brownian Escherichia coli Klebsiella
Testing: determine if an organism is the organisms movement, the pneumonia
Hanging drop motile. An organism must change in organisms
possess flagella to be motile. position with appear quite
respect to each active remain in
other often the same
darting across relative position
the field to other
organisms or
debris in the
field
Motility These tests are used to Motile Nonmotile Escherichia coli Klebsiella
Testing: determine if an organism is organisms will organisms pneumonia
Semisolid motile. An organism must spread out into remain at the
agar deep possess flagella to be motile. the medium site of
from the site of inoculation
inoculation
MRS Broth This test is used to determine Gas production No gas Leuconostoc Pediococcus sp.
whether an organism forms indicated by a production spp.
gas during glucose bubble in the
fermentation. Gas is produced Durnham tube
by some Lactobacillus spp.
and Leuconostoc spp.
MUG Test Escherichia coli produces the Electric blue Lack of Escherichia coli Pseudomonas
enzyme beta D-glucorinidase, fluorescence fluorescence aeruginosa
which hydrolyzes beta-D-
glucopyranosid-uronic
derivatives to aglycons and D-
glucuronic acid. The substrate
4-methylumbelliferyl moiety,
which fluoresces blue under
long wavelength ultraviolet
light
Nitrate This test is used to determine EXPECTED RESULTS NO3+, no gas: Acinitobacter spp.
Reduction the ability of an organism to The nitrate reduction test is read for Escherichia coli
reduce nitrate. The reduction the presence or absence of three NO3+, gas:
of nitrate to nitrite is metabolic products: gas, nitrate Pseudomons
determined by adding (NO3), and nitrite (NO2) aerugenosa
sulfanillic acid and alpha-
naphthylamine. The sulfanillic
acid and nitrite react to form a
diazonium salt the couples
with alpha-naphthylamine to
produce a red, water soluble
azo dye.
Nitrite This test is used to determine No color change The broth Alcaligenes Alcaligenes
Reduction whether an organism can to red 2 minutes becomes red faecalis piechaudii
reduce nitrites to gaseous after the after the
nitrogen or to ther compounds addition of addition of the
containing nitrogen. reagents and gas reagents. No gas
production production is
observed in the observed.
Durham tube.
ONPG This test is used to determine Yellow Clear Escherichia coli Salmonella
the ability of an organism to typhimurium
produce β-galactosidase, an
enzyme that hydrolyzes the
substrate ONPG to form a
visible (yellow) product,
ortho-nitrophenol.
Optochin This test is used to determine Zone of No zone of Streptococcus Streptococcus
the effect of optochin inhibition is inhibition pneumonia mitis
(ethylyhydrocupreine 14mm or greater
hydrochloride) on an in diameter,
organism, Optochin lyses with 6-mm disk
pneumococci (positive test),
but alpha-streptococci are
resistant (negative test).
Oxidase Test To determine the presence of Development of Absence of Neisseria Escherchia coli
(Kovac’s bacterial cytochrome oxidase a dark purple color gonorrhea
method) using the oxidation of the color within 10
substrate tetramethyl-p- seconds
phenylenediamine
dihydrochloride to indophenol,
a dark purple-colored end
product. A positive test
(presence of oxidase) is
indicated by the development
indicates a negative test and
the absence of the enzyme.
Phenylalanine This test is used to determine Green color Slant remains Proteus vulgaris Escherichia coli
Deaminase the ability of an organism to develops on original color
 oxidatively deaminate salnt after ferric after the
phenylalanine to chloride is addition of
phenylpyruvic acid. The added. ferric chloride.
phenylpyruvic acid is detected
by adding a few drops of 10%
ferric chloride; a green-colored
complex is formed between
these two compounds.
PYR Test The PYR test is predominately Bright red color No color change Enterococcus Streptococcus
used in the identification within 5 minutes or an orange faelcalis mitis
schemes for gram-positive color
cocci. Presence of the enzyme
L-
pyrroglutamylaminopeptidase
that hydrolyzes the L-
pyrrolidonyl-β-naphthylamide
(PYR) substrate to produce a
β-naphthylamine. The β-
naphthylamine is detected in
the presence of N, N-
methylaminocinnaldehyde
reagent by the production of a
bright-red colored product
Pyruvate This test is used to determine Indicator No color Enterococcus Enterococcus
Broth the ability of an organism to changes from change; yellow- faecalis faecium
utilize pyruvate. This test aids green to yellow green indicates a
in the differentiation between weak reaction.
Enterococcus faecalis
(positive) and Enterococcus
faecium (negative)
Salt Tolerance This test is used to determine Vivible turbidity No turbidity and Enterococcus Streptococcus
the ability of an organism to in broth with or no color change faecalis mitis
grow in high concentration of without color
salt. It is used to differentiate change from
enterococci (positive) from purple to yellow
non-enterococci (negative).
A heart infusion broth
containing 6.5 % Na Cl is used
as the test medium. This broth
also contains a small amount
of glucose and bromcresol
purple as the indicator for acid
production.
Spot Indole This test is used to determine Development of No color Escherichia coli Enterobacter
Test the presence of the enzyme a blue color development or cloacae
tryptohanase. Tryptophanase within 20 slightly pink
breaks down tryptophan to seconds color
release indole, which is
detected by it’s ability to
combine with certain
aldehydes to form a colored
compound. For indole-positive
bacteria, the blue-green
compound formed by the
reaction if indole
cinnamaldehyde is easily
visualized. The absence of
enzyme results in no color
production (indole negative).
Urea The test is used to determine Change in color No color change Proteus vulgaris Escherichia coli
Hydrolysis the ability of an organism to of slant from the ( agar slant and
Christensen’s produce the enzyme urease, light orange to butt remain light
Method which hydrolyzes urea. magenta orange )
Hydrolysis of urea produces
ammonia and CO2. The
 formation of ammonia
alkalinizes the medium, and
the pH shits is detected by the
color change of phenol form
light orange at pH 6.8 to
magenta pH 8.1
X and V Members of the genus Growth around Growth over the H. infuenzae H. aphrophilus
Factor Test Haemophilus require XV disks only entire surface of will show a halo will grow over the
necessary growth factors in shows the agar growth around entire surface of
vitro. Some Haemophilus spp. requirement for indicates no XV disk; the rest the plate. Neither
require X factor (hemin) alone, both factors. requirement for of the agar X nor V nor XV
V factor (NAD, nicotinamide- Growth around either X or V surface will factors are
adenine dinucleotide) alone, or V disk, no factor. show no growth. necessary for
a combination of both growth around H. growth.
the X disk, and parainfluenzae
light growth will show a halo
around the XV growth around
disk shows a V the XV and V
factor disks.
requirement

PLATING MEDIA FOR ROUTINE BACTERIOLOGY

MEDIUM PRIMARY PURPOSE

Bile Esculin Agar (BEA) Differential isolation and presumptive identification of group D streptococci
and enterococci
Bile Esculin azide agar with vancomycin Selective and differential for cultivation of vancomycin-resistant and
enterococci from clinical and surveillance specimens
Blood agar Cultivation of infectious microorganisms, determination of hemolytic
reactions
Bordet-Gengou agar Isolation of Bordetella pertussis
Buffered charcoal-yeast extract (BYCE) agar Enrichment for Legionella spp.
Buffered charcoal-yeast extract (BYCE) agar Enrichment and selective for Legionella spp.
with antibiotics
Campy-blood agar Selective for campylobacter spp.
Campylobacter thioglycollate broth Selective holding medium for recovery of Campylobacter spp.
Cefoperazone, vancomycin, amphotericin Selective medium for isolation of Campylobacter spp.
(CVA) medium
Cefsulodin-irgasan-novobiosin (CIN) agar Selective for Yersinia spp.;maybe useful for isolation of Aeromonas spp.
Chocolate agar Cultivation of Haemophilus spp. and pathogenic Neisseria spp.
Columbia colistin- nalidixic acid (CAN) agar Selective isolation of gram-positive cocci
Cytine-tellurite blood agar Isolation of C. diphtheria
Eosin methylene blue (EMB) agar (Levine) Isolation and differentiation of lactose-fermenting and non-lactose
fermenting enteric bacilli
Gram-negative broth (GN) Selective (enrichment) medium for enteric pathogens
Hektoen enteric (HE) agar Differential, selective medium for isolation and differentiation of
Salmonella and Shigella spp. from other gram-negative enteric bacilli
MacConkey agar Isolation and differentiation of lactose-fermenting and non-lactose
fermenting enteric bacilli
MacConkey sorbitol agar For the selection and differentiation of E.coli O157:H7 in stool specimens
Manitol salt agar Selective isolation of staphylococci
New York City (NYC) agar Selective for N. gonorrheae
Phenylethyl alcohol (PEA) agar Selective isolation of gram-positive cocci and anaerobic gram negative
bacilli
Regan Lowe Enrichment and selective medium for isolation of B. pertussis
Salmonella-Shigella (SS) agar Selective for Salmonella and Shigella spp.
Schaedler agar Nonselective medium for the recovery of anaerobes and aerobes
Selenite broth Enrichment isolation of Salmonella spp.
Skirrow agar Selective for Campylobacter spp.
Streptococcal selective agar (SSA) Selective for S. pyogenes and S. agalactiae
Tetrathionate broth Selective for Salmonella and Shigella spp.
Thayer-Martin Agar Selective for N. gonorrhea and N. meningitidis
Thioglycollate broth Supports growth for anaerobes, aerobes, microcephallic and fastidious
microorganisms
Thiosulfate citrate-bile salt sucrose (TCBS) agar Selective and differential for Vibrio spp.
Todd-Hewitt broth supplemented with Selective and enrichment for S. agalactiae in female genital specimens
antibiotics
Tryticase soy broth Enrichment broth used for subculturing various bacteria from primary agar
plates
Xylose lysine desoxycholate (XLD) agar Isolation and differentiation of Salmonella and Shigella spp. form other
gram-negative enteric bacilli

TYPES OF HOST:

1. DEFINITIVE HOST – the host in which the sexual reproduction of the parasite takes place or in which the most
highly developed form of a parasite occurs.

2. INTERMEDIATE HOST – host in which alternates with the definitive host & in which larval/asexual stages of a
parasite are found.

3. PARATENIC HOST – host in which larval stage of a parasite survives but does not develop further

4. RESERVOIR HOST – host which harbors the parasite and serves as an important source of infection to the other
susceptible host.

*This host is important in the control and parasitic diseases

TYPES OF HYPERSENSITIVITY REACTION

TYPE 1 TYPE II TYPE III TYEPE IV

ANAPHYLACTIC CYTOTOXIC IMMUNE CELL-MEDIATED
COMPLEXES
Immune mediator IgE IgG or IgM (stevens pg IgG or IgM T cells
210)
Immnune mechanism Release mediators of Cytolysis due to Ag-Ab Deposits of Ag-Ab Release of cytokines
mast cells and basophils and C’ complexes
Examples Anaphylaxis Transfusion reaction Serum sickness Contact dermatitis
Hay fever AIHA Arthus reaction Tuberculin test
Food allergies HDN SLE pneumonitis
Asthma

UREA CLINICAL SIGNIFICANCE

DECREASED INCREASED
 Decreased protein PRERENAL RENAL POST RENAL
intake  congestive heart  acute and chronic  urinary tract
 Severe liver disease failure renal failure obstruction (via renal
 Severe vomiting and  shock, hemorrhage  glomerular nephritis calculi, tumors of the
diarrhea  dehydration  tubular necrosis prostate or bladder)
 pregnancy  increased protein
catabolism
 corticosteroid
therapy
MAJOR CLASSES OF HUMAN PLASMA LIPOPROTEINS: CHEMICAL COMPOSITION

Protein (%) Cholesterol (%) Cholesteryl Triglyceride (%) Phospholipid (%)

Ester(%)
Chylomicrons 1-2 1-3 2-4 80-95 3-6
(exogenous)
VLDL 6-10 4-8 16-22 45-65 15-20
(endogenous)
IDL Intermediate between VLDL and LDL
LDL 18-22 6-8 45-50 4-8 28-24
HDL 45-55 3-5 15-20 2-7 26-32

MAJOR ENZYMES OF CLINICAL SIGNIFICANCE

ENZYME CLINICAL SIGNIFICANCE

1. Acid Phosphatase (ACP) Prostatic carcinoma
2. Alanine aminotransferase (ALT) Hepatic Disorder
3. Aldolase (ALD) Skeletal muscle disorder
4. Alkaline phosphatase (ALP) Hepatic disorder
Bone disorder
5. Amylase (AMS) Acute pancreatitis
6. Angiotensin-converting enzyme (ACE) Blood pressure regulation
7. Aspartate aminotransferase (AST) Myocardial infarction
Hepatic Disorder
Skeletal muscle Disorder
8. Chymotrypsin (CHY) Chronic pancreatitis insufficiency
9. Creatinine kinase (CK) Myocardial infarction
Skeletal muscle disorder
10. Elastase-1 (E1) Chronic pancreatitis insufficiency
11. Glucose-6-phosphate dehydrogenase (G-6-PD) Drug-induced hemolytic anemia
12. Glutamate dehydrogenase (GLD) Hepatic Disorder
13. γ-Glutamyltransferase (GGT) Hepatic Disorder
14. Glutathione-S-transferase (GST) Hepatic Disorder
15. Glycogen phosphorylase (GP) Acute myocardial infarction
16. Lactate dehydrogenase (LDH) Myocardial infarction
Hepatic disorder
Hemolysis
Carcinoma
17. Lipase (LPS) Acute Pancreatitis
18. 5’-Nucleotidase Hepatic disorder
19. Pseudocholinesterase (PChE) Organophosphat poisoning
Genetic variants
Hepatic disorder
20. Pyruvate Kinase (PK) Hemolytic anemia
21. Trypsin Acute pancreatitis

CONDITIONS AFFECTING TOTAL LACTATE DEHYDROGENASE

PRONOUNCED ELEVATION MODERATE ELEVATION SLIGHT ELEVATION

(5 OR MORE TIMES NORMAL) (3-5 TIMES NORMAL) (UP TO 3 TIMES NORMAL)
 Megaloblastic anemia  Myocardial infarction  Most liver diseases
 Widespread carcinomatosis,  Pulmonary infarction  Nephrotic syndrome
especially hepatic metastases  Leukemias  Hypothryoidism
 Systemic shock and hypoxia  Infectious mononucleosis  Cholangitis
 Hepatitis  Delirium tremens
 Renal infarction  Muscular dystrophy
Cushing’s disease: increase in cortisol production caused by excessive development and activity of the pituitary gland

Cushing’s syndrome: increase in cortisol production as a result of tumors which produce either excessive ACTH or cortisol

INFLUENCE OF ACUTE ETHANOL INGESTION ON ETHANOL LEVELS & BEHAVIOUR

BLOOD ALCOHOL (% w/v) SIGNS AND SYMPTOMS

0.01-0.05 No obvious impairment, some changes observable on performance testing
0.03-0.12 Mild euphoria, decreased inhibitions, some impairment of motor skills
0.09-0.25 Decreased inhibitions, loss of critical judgement, memory impairment, diminished
reaction time
0.18-0.30 Mental confusion, dizziness, strongly impaired motor skills (staggering, slurred
speech)
0.27-0.40 Unable to stand or walk, vomiting and impaired consciousness
0.35-0.50 Coma and possible death

Whiskey Blood concentration Influence

(ounces)
1-2 10-50 mg/dL (2.2-10.9 mmol/L) None to mild euphoria
3-4 50-100 mg/dL (10.9-21.7 mmol/L) Mild influence on stereoscopic vision and dark
or greater than 100 mg/dL (21.7 mmol/L) adaptaion
Legally intoxicated
4-6 100-150 mg/dL (21.7-32.6 mmol/L) Euphoria; disappearance of inhibition; prolonged
reaction time
6-7 150-200 mg/dL (32.6-43.4 mmol/L) Moderately severe poisoning; reaction time greatly
prolonged; loss of inhibition and slight
disturbances in equilibrium and coordination
8-9 200-250 mg/dL (43.4 -54.3 mmol/L) Severe degree of poisoning; disturbances of
equilibrium and coordination; retardation of the
thought processes and clouding of consciousness
10-15 250-400 mg/dL (54.3-86.8 mmol/L) Deep, possibly fatal coma

TOXIC AGENTS

Alcohol 1. Ethanol (grain alcohol)

- CNS -most common abused drugs
depressant 2. Methanol (wood alcohol)
3. Isopropanol (rubbing alcohol)
4. Ethylene glycol (1,2-ethanediol)
Carbon Monoxide
Cyanide
Metals 1. Arsenic
2. Cadmium
3. Lead
4. Mercury
5. Aluminum
6. Iron