MODULE 6 MICROBIOLOGY

RIZZA A. CALUAG

Eukaryotes Prokaryotes
• A prokaryotic chromosome is usually a circular
molecule (an exception is that of the bacterium Borrelia
Protists, fungi,
plants, animals
Examples Bacteria/archaea burgdorferi, which causes Lyme disease).
• Even without a real nucleus, the DNA is condensed in a
nucleoid.
Real nucleus NUCLEUS No real nucleus • Prokaryotes can carry extrachromosomal DNA
elements called plasmids, which are usually
Linear DNA Circular circular. Plasmids can carry additional functions, such as
antibiotic resistance.
40s + 60s = 80 RIBOSOMES 30s + 50s = 70
3

Obligate • really need O2 • Leptospira

Live in extreme environments aerobes • TB (upper lobes of lungs:
Halophiles: HIGH salt concentration highly perfused w/ O2)
Thermophiles: HIGH temperature (>6OdegC) • Pseudomonas
Barophiles: HIGH pressure environment Facultative • uses O2 when present but
Methanogens: HIGH methane environment aerobes survives without O2
• can live in anerobic envi coz
Temperature can perform anaerobic
Psychrophilic – cold temperature (~ -20degC) glycolysis
Mesophilic – room temperature Aerotolerant • more anaerobic
Thermophilic – hot temperature (>60degC) (e.g. C. jejuni) • utilize anaerobic glycolysis
but doesn’t die in O2
pH • bet. aero & anaero
Basophile (basic) Obligate • die when O2 is present • ABC – Actinomyces,
e.g. Vibrio, Campylobacter, curve MO like Leptospira anaerobes Bacteroides, Clostridium
Mesophile (neutral) Capneophile • loves CO2 • Campylobacter (dysentery,
Acidophile (acidic) acute gastroenteritis)
e.g. fungi (loves wet and acidic) Microaerophile • needs only 5% O2 • Campylobacter
 Capsulated bacteria–
BACTERIAL TOXINS Some Killers Have Pretty Nice Capsule
• Streptococcus pneumoniae
Endotoxins Exotoxins
• Klebsiella pneumoniae
G- G+, G-
• Haemophilus influenzae
Lipopolysaccharide More virulent
(w/ Lipid A) • Pseudomonas aeruginosa
Heat stable Heat-labile • Neisseria meningitidis
E.g. Meningococcemia E.g. Tetanus pasmin, • Cryptococcus neoformans
Botulinum toxin

Giemsa, Wright and Silver Spirochete

Gram + Stain
Streptococcus Welch Capsule Stain
Staphylococcus
Malachite Green Spores
Listeria Mycobacterium
Bacillus Spirochetes Fuelgen Nuclei
Corynebacterium Mycoplasma
M. Fadyean B. anthracis
Clostridium

Gram - Lowenstein Medium

Modified Thayer Martin
M.Tuberculosis
Neisseria
Neisseria Vibrio
Campylobacter Haemophilus Mac Conkey Agar G-
Chlamydia Pseudomonads
Rickettsiae Helicobacter Fletcher’s Media Leptospira interrogans
Legionella
Chocolate Agar Haemophilus influenzae

COCCI Novobiocin test to distinguish coagulase +/-

Staphylococci :

• S. epidermidis stays away from the novobiocin

disc like an epidemic.
• Therefore, epidermidis is sensitive to
novobiocin.
• S. saprophyticus, much friendlier, comes up to the
disc and says, "'sap?'” [short for "whassup?”]
• Therefore, saprophyticus is novobiocin
resistant. It’s okay to be exposed.
 DIARRHEA
Streptococci: Quellung reaction
EPEC Infant and Children diarrhea
ETEC Traveller’s Diarrhea
EHEC Bloody Diarrhea
"Quell-lung" Shigella dysenteriae Dysenteric Diarrhea
Quell: Capsules swell [+ve test] Vibrio cholerae Rice water diarrhea

Lung: S. pnuemonia Campylobacter jejuni Bloody loose diarrhea

· You get pneumonia in your lung. Bacillus Cereus Starchy Diarrhea

Giardia lamblia Non-bloody Diarrhea
Cryptosporidium and Isospora Severe diarrhea among AIDS Px

2 Forms of Fungi
YEAST MOLD
Filamentous (w/ Nonfilamentous
hyphae) (do not have hyphae)
Unicellular Multicellular
Live at body Temp Live at room Temp
(37oC) (25oC) [outside the
body]

CUTANEOUS MYCOSES Cestodes

TINEA LOCATION CAUSATIVE AGENT CLINICAL MANIFESTATION TREATMENT
Taenia solium • Neurocystecorcosis   DOC: Praziquantel
Tinea pedis (Athlete’s Feet (pork tapeworm) seizures
foot) • Mild intestinal Sx
Taenia saginata • N/V/D, abdominal pain  DOC: Praziquantel
Tinea cruris (Jock itch) Groin area (beef tapeworm)
Diphyllobothrium latum • Vit B12 deficiency  DOC: Praziquantel
Tinea unguium Nails (fish tapeworm) anemia (pernicious
(Onychomycosis) anemia)
Hymenolepsis nana • Abdominal pain, diarrhea  DOC: Praziquantel
Tinea manuum Hands (dwarf tapeworm)
Tinea corporis Trunk Echinococccus  DOC: Albendazole
Tinea barbae Beard area/chin granulosus
(dog tapeworm)
Tinea capitis Scalp/head
 NEMATODES
Trematodes – flukes DISEASE PARASITE OTHER NAME

DISEASE CAUSATIVE AGENT CLINICAL TREATMENT Anisakiasis Anisakiasis worm/ Phocanema

MANIFESTATION
Ancylostomiasis Ancylostoma duodenale Old World hookworm
Schistosomiasis Schistosoma japonicum Katayama Fever DOC: Praziquantel
(Oriental Blood Fluke) Chronic Liver Ascariasis Ascaris lumbricoides Giant Intestinal Roundworm
Pathology Capillariasis Capillaria philippinensis
Lung Fluke Paragonimus westermani Mimics TB ~ lung DOC: Praziquantel Dracunculiasis Dracuncuculus medinensis Guinea/serpent/dragon worm
(lung fluke) cavitation Enterobiasis/oxyuriasis Enterobius vermicularis Pinworm/seatworm
• Hermaphrodite Bancroftian filariasis Wuchereria bancrofti Bancroft’s filaria
Malayan Filariasis Brugia malayi Malayan filarial worm
Liver Fluke • Chlonorchis sinensis • Inflammation of DOC: Praziquantel Filariasis, occult Dirofilaria species Heartworm
(Chinese liver fluke) lymph nodes Loiasis/fugitive swelling/calabar swelling Loa loa Eye worm
• Opisthorchis viverrini • Hepatomegaly
(Southeast Asian liver Necatoriasis Necator americanus New world Hookworm
fluke) American Hookworm
Onchocerciasis Onchocerca volvolus Nodular/Blinding worms
Strongyloidiasis/ Strongyloides stercoralis Threadworm
MOT: Undercooked fish Cochin-china diarrhea
meat
Trichinosis/Trichinellasis Trichinella spiralis Trichina worm
Trichuriasis/trichocephaliasis Trichuris trichura Whipworm

PROTOZOA TRANSMISSION MORPHOLOGY CLINICAL DIAGNOSIS African -Tse tse fly -Trypomastigote: African -visulaize -Suramin
Trypanosome bite motile, Sleeping trypomastigote -Melarsoprol
Entamoeba Fecal-oral -oocyst -asymptomatic: -fecal exam
-Trypanosoma -contaminated extracellular form Sickness in blood, spinal,
histolytica -trophozoite: motile carriage -serology
-not all species bulls-eyed shaped -bloody diarrhea -abdominal CT rhodesiense BT -Trypomastigote fluid or lymph
of Entamoeba nucleus w/ RBC in -liver abscess scan -Trypanosoma and epimastigote nodes
are pathogenic cytoplasm -AMEBIASIS gambiense in tse tse fly -serology

Giardia lamblia Fecal-oral -oocyst Foul smelling -fecal exam

-trophozoite: greasy diarrhea and -commercialized
American -Kissing Bug: -Trypomastigote: Chaga’s -visualize -Nitrofurtimox
flagellated abdominal gassy immunoassay kit
distention Trypanosome defecates on motile disease trypomastigotes -Benznidazole
- GIARDIASIS -Trypanosoma human skin extracellular form in blood
Trichomonas Sexually -no cyst stage -painful vaginal -examination of cruzi while feeding -Amastigote: non -xenodiagnosis
vaginalis fomites -trophozoite: itching vaginal discharge - motile,
flagellated -burning on Contaminated intracellular
urination BT -Trypomastigote
-yellow green and epimastigote:
malodorous frothy in kissing bug
vaginal discharge
-TRICHOMONIASIS/
VAGINITIS/PINGPO
NG INFECTION

• CHLOROQUINE: DOC IN ACUTE ATTACK, also used in

Important Tissue or Blood Protozoans
PROPHYLAXIS
Malaria Female anopheles -trophozoites Malaria
-Plasmodium falciparum mosquito -schizonts • Primaquine: eliminates the exoerythrocytic forms in the
-Plasmodium vivax -gametocytes
-Plasmodium ovale
liver(relapsing malaria) : radical cure
-Plasmodium malariae • Mefloquine- chemoprophylactic drug of choice in chloroquine
resistance
• Proguanil (100 mg) and atovaquone (250 mg)– malarone 
both for prophylaxis and treatment of multi-drug resistant
falciparum malaria
• Pyrimethamine-sulfadoxine  presumptive treatment in
case of malarial breakthrough
• *no drug regimen can ensure prevention of malaria
 Leishmaniasis The 21 Virus Families
• Leishmania tropica, Leishmania chagasi, Leishmania DNA viruses
major, Leishmania braziliensis, Leishmania donovani (HHAPPPPy)
• Zoonotic disease Hepadnaviridae
• Sandfly Herpesviridae
Adenoviridae
Leishmania donovani
Visceral leishmaniasis, Kala-azar- Dumdum fever
Poxviridae
Leishmania braziliensis
Mucocutaneous leishmaniasis
Papovaviridae
Parvoviridae
Tx: antimonial stibogluconate (Stibofen ®)
Polyomaviridae

The 21 Virus Families

RNA viruses Retroviridae
(PRAyFOR PRC)TB Paramyxoviridae
Picornaviridae Rhabdoviridae
Reoviridae Caliciviridae
Astroviridae Coronaviridae
Arenaviridae Togaviridae
Flaviviridae Bunyaviridae
Filoviridae
Orthomyxoviridae

NAME CLASSIFICATION TRANSMISSION CLINICAL TREATMENT

DNA Viruses RNA Viruses Hepatitis A *Picornaviridae Fecal-oral Acute viral hepatitis -pooled immune
serum globulin
Double EXCEPT Parvovirus Single EXCEPT Reoviridae
-supportive care
stranded stranded and Rotaviridae new HAV vaccine
Icosahedral EXCEPT Helical/ EXCEPT Rhabdo  Hepatitis B *Hepadna viridae -BT -Acute viral hepatitis -Hepa B vaccine
-needle sticks -Fulminant hepatitis -alpha-interferon
Poxvirus (complex) icosahedral helical bullet -Lamivudine
-sexual -Chronic hepatitis
-placental Complications:
*Naked EXCEPT Enveloped EXCEPT Reoviridae, -hepatocellular CA
-cirrhosis
Hepadnavirus, Herpes Picornaviridae, Hepatitis C Acute viral hepatitis
*probably a Flavivirus -alpha interferon
virus, Pox virus Calciviridae (PRC) -Ribavirin
(enveloped)
Replicate in EXCEPT Replicate in EXCEPT Influenza Hepatitis D -incomplete RNA virus -coinfection w/ HBV Control of HBV
-superinfection infection
nucleus Pox virus cytoplasm and Retrovirus Complication
(nucleus) -fulminant hepatitis
Linear DNA EXCEPT -cirrhosis
Hepadnavirus, Hepatitis E *probably a Calici virus Fecal-oral Hepatitis (like A)
Hepatitis G Flavivirus -transfusion Has not been
Papilloma virus, -needle sticks conclusively shown to
Polyoma virus cause liver disease
Coronaviridae
• Corona virus –
common colds; NEW VIRUSES
SARS
Retroviridae
• Human
immunodefficiency
virus (HIV) - AIDS

OTHERS: Immunotherapy
Active Immunization
ADENOVIRIDAE- childhood URTI Live Virus Killed virus Virion subunit Viral polypeptides DNA Vaccines
Vaccines vaccines vaccines

Measles Polio (Salk) Adenovirus HBV HIV

OTHER VECTOR-BORNE VIRAL DISEASE: Mumps
Rubella
Rabies
Influenza
Influenza

REOVIRIDAE- COLORADO TICK FEVER CAUSED Chicken pox

Polio (Sabin)
HAV

BY COLTIVIRUS Yellow fever

Passive Immunization
TOGAVIRIDAE- CHIKUNGUNYA
HAV, HBV
BUNYAVIRIDAE- SANDFLY FEVER CAUSED BY Measles
Rabies
PHLEBOVIRUS RSV
VZV

Relapsing Fever Body Louse

FEVER
Leishmaniasis Phlebotomus/sand fly
5- day fever/ Trench Fever Bartonella quintana
Rabbit Fever Fracinsella tularensis
American Sleeping Reduviid bug/Kissing Bug
sickness Q-Fever Coxiella burnetti
Cat-scratch Fever Bartonella henselei
African Sleeping Sickness Tsetse fly
Relapsing Fever Borellia recurrentis
Babesiosis Scarlet Fever Streptococcus pyogenes
Ixodes Tick Typhoid Fever Salmonella typhi
O. Vulvulus Black flies/simulium fly Undulant/Malta Fever Brucella mellitensis

Lyme Disease Small Ticks

 VIRAL VACCINES
LIVE ATTENUATED KILLED
Stimulate both humoral and Only initiate humoral
cell-mediated immunity
Relatively dangerous to
administer
E.g. small pox, yellow E.g. rabies, influenza, salk
fever, chicken pox, sabin, polio, HAV
polio, MMR (measles, PUBLIC HEALTH
mumps, rubella)

SPECIFIC IMMUNITY
• Aka adaptive immunity
• (2) types:
1. Natural
• Naturally acquired active immunity – host produces its
own antibody after recovery from a disease
• Naturally acquired passive immunity – mother to fetus
(maternal)
2. Artificial (by vaccination)
• Artificially acquired passive immunity - an immediate,
but short-term immunization provided by the injection of
antibodies, such as gamma globulin, that are not
produced by the recipient’s cells.
• Artificially acquired active immunity - microbe, or parts
of it, are injected into the person before they are able to
take it in naturally. If whole microbes are used, they are
pre-treated, attenuated vaccines.

QUALITY CONTROL AND

ASSURANCE
SX SIZE
PACOP QUESTION
ULTRAMICROANALYSIS <1mg
MICROANALYSIS 1-10mg The type of alkaliodal assay where the total alkaloid is
SEMI-MICROANALYSIS 10-100mg determined.
MACROANALYSIS 100mg-1000mg A. Ultimate
B. Specific
EXTENT OF DETERMINATION
C. Proximate
PROXIMATE TOTALITY/GROUP D. Extraction
ULTIMATE SPECIFIC CONSTITUENT/ SINGLE
CHEM.SP.

ACIDIMETRY
Analyte: Free Base Aqueous Non - aqueous
Titrant: Acid

VS Hydrochloric acid Perchloric acid in

glacial acetic acid

Sulfuric acid Perchloric acid, HBr

Primary Standard Anhyd. Sodium carbonate Potassium biphthalate

Secondary Standard Sodium/Potassium hydroxide --------------------

Examples:
• Direct NaOH, NaHCO3, Na salicylate, Methacholine Cl
Caffeine, NaCO3

• Residual NH4Cl, Methenamine,

ZnO, Potassium sodium
tartrate, tartrate

ALKALIMETRY
Analyte: acid Aqueous Non – aqueous
Titrant: free base
VS Sodium hydroxide Sodium methoxide in
ethanol or Toluene,

Potassium hydroxide Li Methoxide

Primary Standard Potassium biphthalate Benzoic acid

Secondary Standard Hydrochloric acid in Sulfuric acid

Examples:
• Direct Phosphoric acid, Hydrochloric Phenytoin
acid, Tartaric acid W/ R agent= RED
W/ O agent= GREEN/BLUE
• Residual Aspirin
 DIAZOTIZATION COMPLEXIOMETRY
• NOT IODOMETRIC METHOD:
• assay for sulfonamide drugs and other compounds
containing Arylamines
• VS: 0.1M 0.1 M sodium nitrite
• Refernce std: K2Cr2O7
• Endpoint: blue
PACOP Q:
Which of the following volumetric solutions is used in
diazotization analysis of sulfa drugs?
0.1 M sodium nitrite

PRECIPITATION METHOD PRECIPITATION METHOD

Manifestati
METHOD Manifestation VS Indicators 1 std Example METHOD VS Indicators 1 std Example
on
GAY- cessation of Silver NaCl NaCl
formation of LUSAAC ppt. Nitrate
VOLHARD insoluble Ferric
Ammonium silver Aminophylline, Sodium
(residual) colored Ammonium appearance K Organic N
complex Thiocyanate nitrate NaCl LEIBIG Teraphenyl Bromophenol
Sulfate of turbidity biphthalate compounds
(colored ppt) boron
Adsorption Phenylephrine
Change in Silver indicator HCl
FAJANS NaCl
formation of ppt Nitrate Ex. DCF, Tubocurarine
Potassium HCl
MOHR 2nd Silver Nitrate NaCl SLS Eosin Y, TEE
precipitate chromate

APPROXIMATE TEMPERATURE EQUIVALENTS

Very Dull Red Heat 500C-550C

Dull Red Heat 550C-700C

Bright Red Heat 800C- 1000C

Yellow Red Heat 1000C-1200C

White Heat 1200C- 1600C

 BIOLOGICAL METHODS BIOLOGICAL REACTIVITY TESTS
DIGITALIS PIGEON (IN VIVO)
HCG FEMALE RATS
INSULIN RABBIT(HPLC) TEST ANIMAL USED NOTES
HEPARIN SHEEP BLOOD
GLUCAGON CAT PLASMA Systemic Albino mice for elastomeric material
Injection Test esp. plastics
OXYTOCIN CHICKEN
PROTAMINE SO4 SHEEP BLOOD Intracutaneous Albino rabbits for elastomeric material
PLASMA
VASOPRESSIN MALE RAT Test esp. plastics
COD LIVER OIL RACHITIC RAT
CORTICOTROPIN RAT Eye Irritation Test Albino rabbits for medical devices &
PARATHYROID DOGS ophthalmic containers
TUBOCURARINE INJ. RABBIT(HPLC) HORMONE

METOCURARINE INJ. RABBIT (HPLC) Implantation Test Rabbits for material in direct
contact w/ tissue

LIMIT TEST ASSAY METHODS

SPECTROPHOTOMETRY Vitamin A
As DEC Vitamin K (635 nm)
Cl AgNO3 Vitamin B12 (361nm)
Vitamin B3 (450 nm)
SO4 BaCl2
Steroids (525 nm)
HEAVY H2S
HPLC VIT. D, B6, B9, PEN, CEPHA
METALS
Fe NH4SCN GC BARBITURATES, VIT.E
Pb AMMONIA CYANIDE, FLUOROMETRY VIT. B1,B2
AMMONIUM CITRATE,
L.plantarum VIT B3, B5, B12
HYDROXYLAMINE, HCl,
DITHIZONE RABBIT BLOOD INSULIN ACTIVITY
SUGAR METHOD