BACTERIOLOGY QUESTIONS MIDTERMS

BACTERIAL CULTIVATION (LAB)

-TOPICS (6)
SOURCES OF METABOLIC ENERGY:
^________________ - uses substrate phosphorylation, an enzymatic process in which a
pyrophosphate bond is donated directly to ADP by a phosphorylated metabolic intermediate
^(next dito yung the formula chuchu ba un?)
^Most sugar that are being used for fermentation: (4)
^A chemical reduction of an oxidant (electron acceptor) through a specific series of electron
carriers in the membrane of prokaryotes establishes the proton motive force across the
bacterial membrane.
^similar to the oxidant-reduction of respiration, _____________ oxidant and reductant are
created photo chemically by light energy absorbed by pigments in the membrane. The product
is __
NUTRITION
^_______________ requirement of ___________ (organisms that do not require organic
nutrients for growth) that depends on photosynthesis
>_______________ organisms that use inorganic substrate (H+, HSO32-, CO2)
for carbon source
>_______________ requires organic carbon source (CHO, CHONS, AA, etc.) for
growth
>_______________ requires carbon dioxide for growth
^_______________ is needed for the biosynthesis of amino acids and nucleic acids and other
compounds
>_______________ property, unique to prokaryotes, high energy required
process, that is able to break nitrogen-nitrogen triple bonds
>example of ^ as a source: NH3 (ammonia) through Ammonification
^______________ important in the cysteinyl and methionyl side chains of bacterial proteins
and for several coenzymes. ____________________ to sulfate used as source of sulfur. Some
microorganisms can assimilate H2S directly from some media
^______________ required by prokaryotes for ATP production. Nucleic acid biosynthesis, and
coenzymes (NAD,NADP, and Flavin), and phospholipid synthesis, cell wall components (teichoic
acids), some capsular polysaccharide. It is always assimilated as free inorganic phosphate.
^______________ most of this required for enzyme functions
>_____&_____ are found in porphyrin derivatives, magnesium in chlorophyll
molecule, and iron as part of coenzyme of the cytochromes and peroxidase.
>_____ is required for the constituent of gram positive cell wall and
>_____&_____ are essential for integrity of ribosomes
>_______________ membrane proteins that require iron for synthesis for
transport of soluble complexes
^________________ organic compounds which a cell must contain, but these are not
synthesized intracellularly. Most are building blocks of biomolecules such as: (6)
>__________________-the following must be provided: (5) AA-amino acids….
^__________________ prokaryotes require specific pH range for survival, and the range
optimal to some organisms are very narrow. (3)
^__________________ most organisms grow in normal body temperature ( __ ). But there are
different species vary widely in their optimal temperature ranges for growth: (4)
^_________________ discussing the role of oxygen as hydrogen acceptor
>_________________- specifically requiring oxygen as hydrogen acceptor
>_________________- organisms that can live aerobically or anaerobically
>_________________- requiring substance, other than oxygen and hydrogen, and
being sensitive to oxygen inhibition.
^_________________ control of salt concentration and osmotic pressure in the medium is
required
>_____________- are organisms requiring high salt concentration in the
media.
>_____________- are organism that require high osmotic pressure in the
media
SPECIMEN COLLECTION:
GUIDELINES FOR SPECIMEN COLLECTION:
1. It should be collected at the correct anatomic site
 2. It should be collected acute phase of illness (for virus: __-__ days)
3. Before antibiotic therapy or chemotherapy should be collected
> if the px is on antimicrobial, use ____ (pati yung example neto) -to neutralize
antimicrobials
> __________________ be neutralized by addition of PABA (paba stands for?)
>______________ can be counteracted with use of penicillinase to medium
4. Specimen preservatives may be used
>____________- for urine
>________________________________- for stool (ova and parasitic exam)
5. Timing of collection should be considered
>__________________- should be collected during acute diarrheal stage
>__________________- for this isolation, blood is specimen of choice for 1 st week of
infection, ______or______ for 2nd or 3rd week and ______________ for 5th week
6. Specimen should be collected under sterile, aseptic condition
7. Quantity of material must be adequate
8. _________________ or __________ or __________ swabs are recommended for
bacterial culture, ______________ may release toxic fatty acids
9. Specimen must be examined promptly within ____
10. Label the specimen correctly
11. If specimen cannot be processed ASAP, they must be stored in proper temp.
>Refrigerator temp __ degrees Celsius (5 specimens?)
>Room temp ___degrees Celsius ( _________ bacteria)
>Body temp __degrees Celsius (___ or ________)
>Freezing temp ___ to ___ degrees Celsius ( ______ for ______ studies tissues as
specimen)
SPECIMEN COLLECTION: BLOOD
>__________ is usually sterile. Specimen using this is used to evaluate bacteremia, septicemia
and fever of unknown origin
^_______________ presence of bacteria in blood
>___________-normal flora is introduced in the blood
>___________bacteria is sporadically discharged from extravascular
abscesses or infection to the blood
>___________ constant release of bacteria into the blood
^________________ refers to the presence of toxins in the blood
>_________________ multiply faster than rate of phagocytosis
>________________ and systems may be infected
^Blood is collected ___ to ___ minutes before the fever spikes
^Blood culture bottle is used with anticoagulant ( ____ to ____ sodium polyanethol sulfonate)
^SPS is used for:
>prevents clotting
>prevents phagocytosis
>neutralizes bacterial effect of complement
>inactivates antimicrobials
>inhibits some organisms like N. gonorrhea, N. meningitides, Peptostreptococcus
anaerobius and Gardnerella vaginalis
^____________ can be used as anticoagulant for viral cultures
^_________ , _________ should not be used
^Collection: __ to __ sets ( __ set is composed of 2 bottles always) , should be collected from
different sites, __ hour interval within __ hours
^__ to __ mL of blood per set for adults; __ to __ mL per set for children
^Blood cultures are incubated at __ to __ degrees Celsius for __ to __ days
^for _________ spp, blood should be incubated for __ weeks

SPECIMEN COLLECTION: CSF

-Collected through ________ puncture or ___________
-________________ is done between __ & __ OR __ & __
-_______ is normally sterile, this is used to evaluate __________
-______________ may enter through _____________ , _________ may enter through the
___________ cells
-isolates from CSF are usually ___________ and ______________
-Primary culture media used is ___________________
-examined immediately or if incubated for _______ an hour only
-3 tubes are collected:
> 1st tube _____________ and ______________
 > 2nd tube _____________ for ______________
> 3rd tube _____________ for ______________
-if only 1 tube is collected, should be sent first to ______________ , then _____________ then
______________ and _____________
-if _________ are made, this is prepared from ______________ through centrifugation RPM
(revolutions per minute) ______ for ___ to ___ minutes
-______________ and ___________ is used for smears

SPECIMEN COLLECTION: GASTROINTESTINAL

^Composed of _______ , _____________ and _____________
^_________ is one of the choice for gastrointestinal pathogens
^_____________ is collected by inserting swab ___ cm _________________. _______ should
be visible in the swab
^if bacterial pathogen is suspected, __ specimen per day for __ days consecutive should be
collected
^Specimen should not be taken from toilet bowl
^specimen should not be contaminated with urine and water

SPECIMEN COLLECTION: URINARY TRACT

^Includes ____________________, _________________, and _________________
^_______________ urine is ideal
^for ______________ sample, first ___mL is allowed to pass then collect
^Urine specimen must be cultured within __ hour or refrigerated not longer than ___ hours
^____________________ is done on BAP
^Colonies are reported as _________________________ per mL of urine
^significant bacteriuria is _________
SPECIMEN COLLECTION: RESPIRATORY TRACT
^_________________- swabbing in the posterior pharynx, tonsils and inflamed areas
>used for (3)
^_________________- moistened swab in the nose and detects _____________________
^__________________- posterior nasopharynx rotated for __ seconds and detects
______________________ and ___________________
^_______________ - collected in the morning,
>deep cough is ideal,
> ___ morning sputum is recommended for __________________
> detection of ________________________

SPECIMEN COLLECTION: GENITOURINARY

^ used to determine cause of ____________ , ____________ and ______________ as well as
_______________________
^often used to detect ___
^______________________ is susceptible to temperature fluctuations and ____ level of ____
^most ______________ bacteria requires ___________________

SPECIMEN COLLECTION: WOUNDS AND ABSCESSES

^_____________________ -Infection from animal or human bites, burns, ulcers and traumatic
wounds (gunshots)
^_____________________- attributed to bacterial source within the patient, includes cellulitis,
dental infections and septic arthritis
^ applicable for ______ infections, _______________ wounds
^preferred specimen is _______________________
^when using swabs, care must be taken during collection to prevent contamination with normal
flora
MATERIALS USED IN INOCCULATION: (6)
INOCULATION TECHNIQUES: (4)
^______________________ - streaking used for ___________
>_______________ is applied by swabbing a dime-sized area or placing a
drop of liquid specimen into the plate
>STEPS: 1. Streak the mixed culture back and forth in one quarter of the agar.
Flame the loop and proceed
2. rotate the plate nearly 90 degrees, streak again across the second
sector and then Flame the loop
3. rotate the plate nearly 90 degrees, streak again across sector 3.
Flame again
4. after cooling the loop, streak for one last time into the center of
the plate. Flame the loop and incubate the plate in an inverted position.

>GRADING: __ -many, heavy growth; if growth is out to the Q4

__ -moderate growth; if growth is out to the Q3
__ -few or light growth; if growth is out to the Q2
__ -rare; if growth is in the Q1 only
 __________________- dish divided into 4 quadrants, serially streaked
^________________________ - used for urine cultures and tissues from burn victims
>uses __ uL ( ____ ) or __ uL ( _____ ) calibrated
>incubate __ to __ hours and count colonies
>STEPS: 1. Streak a single vertical line
2. go back and cross streak in spiral manner
> reporting:
> __ colony is approximately ______cells
^________________________ - done in tubes or slant agar ( magbigay 2 examples)
>Used for _________________ and _____________
>STEPS:
>_______________________- for uniform culture condition and cultivation
>easier to do that the solid media or semi solid
 >STEPS:

CULTURE MEDIA: TYPES OF MEDIA

According to Consistency: (4)
According to Composition: (3)
According to Purpose: (9)
^______________________- for transport of organisms to prevent drying and disintegration
^ (4 examples)
^______________________- supports the growth of most non- fastidious organisms
^ (3 examples)
^______________________- enhances the growth of one group while inhibiting the growth of
others
^ (4 examples) Alkaline peptone water
^______________________-derived from the simple medium with added nutrients like blood,
serum, peptone and vitamins
^ (2 examples) Blood agar plates, chocolate agar plate
^______________________- selective by either the addition of substance that selectively
favors the growth of particular organism or by addition of bacteriostatic agent which controls
the growth of undesired organisms,---most _____________ are also differential media, thus
referring them as “selective-differential media”
^ (5 examples of selective agents and 3 sa dulo) (mannitol salt agar,
thiosulfate citrate bile salts agar)
^ _______________inhibits G+ bacteria
^ _______________inhibits G+ bacteria
^ _______________prevents spreading of Proteus spp
^ _______________Inhibits G- organisms
^ _______________prevents growth of contaminating organisms
^_____________________- used for Kirby Bauer disk diffusion in AST (Antimicrobial
susceptibility testing)
^_____________________- provides distinct colonial appearance of organisms
^_____________________- medium that contains antibiotic as inhibitor
^_____________________- used for testing the biochemical activities and detect chemicals
produced by the organisms
^ (4 examples) Kligler iron agar, Triple sugar iron agar, sulfide indole
motility
GRAM NEGATIVE COCCI
Topics: (4)
>Clinically significant family are _______________ and _______________
> Both oxidase __
>_______________ is formerly known as _______________
>Pathogenic ________________ are usually fastidious and would grow optimally at ___ to ___
degrees Celsius
> Requires enriched media containing ( 3 examples) which are all found in ______
> All _____________ spp are Catalase __ except for ________________

NEISSERIA GONORRHOEA
^Other name: ______________
^Introduced by: ___________________ in the year of ______
^Gram ___ ________________ (what shape? example)
^_______________ to WBC
^ Oxidase __
^Oxygen Tension: ________________
^_________________ forming
^____________-twitching motility
^ Enrichment media: (2 examples)
^Special structure: ______

CLINICAL SIGNIFICANCE:
>________________________- affects the genitourinary tract
^ acute pyogenic infection in ________________ of the _________ in
females and __________ in males
 ^accompanied by ___________- burning urination and pus-like discharge in
men
^for women, (4 examples of symptoms)
^many gonococcal infections are (asymptomatic or symptomatic?)
^long term effect on female: (3)
^___________________ and __________________ can be seen in sexually
active homosexual and bisexual men
>___________________________ - gonococcal infection that spreads in ___________
^_______________ characterized by hemorrhagic skin eruptions
^may cause ________________ and __________________
>_______________________- ________________ acquired by newborn from gonococcus
infected mother during delivery
^this can result to ______________ of the newborn

VIRULENCE FACTORS (5)

^____________________- attachment of organism to mucous membrane thereby initiating
infection
^____________________- affects antibody formation, leukocyte response and cell mediated
immunity
^____________________- activates the complement pathway of the host producing endotoxin.
Stimulates also the production of Tumor Necrosis Factor that causes cell damage
^_________________- allows the organism to insert itself to the host cell
^_________________- allows the organism to attach to neutrophils and epithelial cells and to
resist effects of antibodies

SPECIMEN: (4)
CULTURE MEDIA: (4)
^Cannot grow on ______ and ________
IDENTIFICATION:
^Gram staining: ______________________ is more significant in male patients since G- cocci in
females can also be _________________ or _________________
^Note of the abundance of ___ (ano to) and amount of _____________ present
^ E.g. of reporting few gram negative diplococci occurring intracellular to
few ___
^CONFIRMATORY TEST: _____________________ - checks the organism to use carbohydrate
with production of acids
^use of ____________________________ with added __ of sugar
^__ result is production of acid indicated by ________ color of the __
indicator ( ___________ )
^N. gonhorrhea ferments ______________

NEISSERIA MENINGITIDES
^Other name: ___________________
^Gram __ ________________ (shape neto)
^_______________ and _________________
^Oxidase __
^Oxygen tension: ______________
^______________ forming
^(motile or nah?)
^Special structure: ____________
>the organism can be carried by __% of the population as normal flora in the _______________
>sero-grouping of the spp. Are based on type of ___________________
>examples of sero groups are: (9)
>___________________ is the portal of entry
>Infections can be transmitted to members of the population that cannot produce
_______________ (and 2 examples)
CLINICAL SIGNIFICANCE: (2)
^_______________________- presence of _______________ in the ____
^_______________________- presence of _______________ in the _____
^__________________________ is fatal
^__________________________ is characterized by recurrent episodes of
meningococcemia, ______________ formation and eventually ____________ development
^______________ formation is due to endotoxin
^______________ is characterized as tiny hemorrhage into the skin which
leads to disseminated intravascular coagulation
^______________________________ fulminant meningococcemia associated with bleeding of
adrenal glands

VIRULANCE FACTORS : (3)

^_______________ - promotes attachment
^_______________- causes petechiae
^_______________ - enables organism to resist ________________ and enables them to cross
the _______________ leading to meningitis

SPECIMEN: (5)
IDENTIFICATION: (3)
^____________________- Same reporting with N. gonorrhea
^______________- _____ is centrifuged to concentrate the specimen. _____ and
_________________ is used as medium
^_________________________ may contain contaminants therefore
____________________ must be used (2 examples)
^ Colonies are usually ___________ , __________, _____________ ,
_______ and sometimes ____________
^Neisseria menigitidis may grow on _____ but not on _____
^Confirmatory test - _________________________test, ferments _____________ and
______________
MORAXELLA CATARRHALIS
>Former name: __________________________ , _______________________
> Gram __ _________________
>______________
>Oxidase __
>Oxygen tension: ___________
>_________________ forming
>(motile or nah?)
>special structure: (2)
>MORAXELLA CATARRHALIS CAN BE MISTAKEN BY NEISSERIA GONORRHOEAE

CLINICAL SIGNIFICANCE: (2)

^____________________- inflammation of the middle ear
>can be cause also by ______________________
^ (8)

IDENTIFICATION: (2)
^______________________: M. catarrhalis does not utilize any _____
^______________________ (while ___________________ is negative)
LESSON 5. GRAM POSITIVE BACILLI
> 4 TOPICS
>____________________SPP- Gram __ bacilli “______________”
>Oxygen tension: ____________/_______________
>Catalase __
>Oxidase __
> Spore forming or nah?
>Motile or nah?
>Special structure: _________________ ( _____________________ )
>___________________- ability to alter their morphology, biological functions or reproductive
models.

^_________________________:Other name: _____________________

^Morphology:__________________
^Arrangement: (3)
^the darker coloration in staining are the _______________________ in the _______________
and not ___________
^Special stain for the metachromatic granules: ________________ and _________________
^(3) distinct cultural types:
^Virulence factor: ________________
^CLINICAL SIGNIFICANCE (2):
>__________________- serious infection that usually affects the mucous membranes of the
nose and throat, Can lead to difficulty in breathing, heart failure, paralysis and death, RARE
condition now in 1st and 2nd world countries due to vaccination in infants…..
^DPT-_____________________ developed by ________________________
^Transmission: ________________ ( _________________ )
^Incubation: __ to __ days (at most __ to __ days)
>___________________-Common occurs on exposed limbs particularly the legs, Characterized
by lesions as vesicles and quickly form multiple ulcers, Classic characteristics of the lesions is
usually covered with an _________________- a hard bluish gray membrane that is slightly
raised
>can be caused also by ___________________________
CULTURAL CHARACTERISTICS: (5)
^______________________- colonies have narrow zone of beta hemolysis
^______________________- enhances the metachromatic granule formation
^______________________- forms colonies that are black color with brown halo
^______________________-gray to black colonies
^______________________- gray to black colonies

(Under cultural characteristics) COLONIAL TYPES OF C. diphtheriae in BAP (3)

^_________________- __ to ___mm colonies; largest colonial type
^_________________- Fried egg appearance (clear colonies with white center) bleach-like odor
on Tellurite medium
^_________________- ___mm colonies, Smallest, appears black colonies with gray borders on
Tellurite medium

DIPTHEROIDS/CORYNEFORM BACTERIA (5)

-___________________-TYPICALLY CAUSES OPPORTUNISTIC INFECTIONS IN
IMMUNOCOMPRIMISED HOSTS
^___________________________- cause infection to bone marrow transplant patients, Can
cause endocarditis, Resistant to number of antibiotics such as: (3), susceptible to: (2)
^___________________________- mostly are normal flora and can cause infections to
immunocompromised hosts, Can cause diphtheria-like infection ( __________________ )
^___________________________- formerly known as C. ovix, Infects horses and rarely on
human
^___________________________- NF of the skin , Nasopharynx and conjunctiva
^___________________________-a.k.a C.___________ , NF of the oropharynx and throat
LISTERIA SPP
>Gram __ bacilli
>Oxygen tension: ___________________
> spore forming or nah?
> intra___________
LISTERIA MONOCYTOGENES
^Gram __ to gram ________ coccobacilli
^can be mistaken as STREPTO ________________________
^Motility: (3)
^can be also found in the: (3)
^____________________ - can tolerate both acidic and salty conditions
^Catalase __
^Oxidase __
^________________hydrolysis __
^CAMP test __
^Hemolysis: _____________________
^Special structure: _____________ and _______________________ ( ______________ is
absent at room temperature )

CLINICAL SIGNIFICANCE:
^ Transmission: _______________________________________ and ___________
>(4)
VIRULENCE FACTORS: (3)
^____________________- contributes to the intracellular mobility
^____________________- a hemolysin
^____________________- prevents phagocytosis
Biochemical Characteristics: (3)
>MVCCHINOU
>GTSM
>MS LC

SPECIMEN OF CHOICE: (3)

CULTURAL CHARACTERISTICS: (3)
>__________________ - umbrella shaped or inverted Christmas tree pattern
>__________________- smooth, clear to gray colonies with narrow band of
beta hemolysis
>__________________- bluish green colonies

ERYSIPELOTHRIX SPP
>Gram __ straight to slightly curved or slender bacilli
> motile or nah?
>can be seen singly in V shaped pairs or in short chains
>they have tendency to _______l_________________
>Oxygen tension: _______________

ERYSIPELOTHRIX RHUSIOPATHIAE
>Gram __
> spore forming or nah?
>Catalase __
>Oxidase __
>Alpha_____________ to _______________ in BAP
>motile or nah?
> primarily considered an ________________________
>can be (3)

CLINICAL SIGNIFICANCE: (2)

^____________________________- occupational hazards to individual handling meat and
poultry,
>”__________________” swelling at the site of organisms inoculation
*________________- mistaken close term to erysipeloid is not caused by E. rhusiopathiae in
human but the genus Streptococcus
^____________________________-presented by a diamond-shaped cutaneous lesion to
animals

VIRULENCE FACTORS: (2)

^____________________- Heat labile encapsulated enzyme
^____________________- bacterial attachment and subsequent invasion into host cell

SPECIMEN OF CHOICE: (3)

BIOCHEMICAL TESTS: (3)
^IHNC
^GMS
^”______________________” growth on semi solid medium

OTHER GRAM-POSITIVE BACILLI: (2)

^________________________
> motile or nah? ____________________
>normal flora of the: (3)
>has a ________ clinical significance
>tolerates __________________________ (can grow at pH __ to __ )
 >cultured on ________________________
>Catalase __

^__________________________
>causes chronic pulmonary infections in immunocompromised patients
>stained___________________________ with _____________________
>characterized by: (3)
>grows in SDA
>inhibited by __________________________ and can grow in any media that
does not contain _____________________
>CUP
MACROSCOPIC EXAMINATION:
TOPICS: (2)

GROSS MACROSCOPIC EXAMINATION:

^(3) -description of the shape and edge of colonies
>FORM (6)
>MARGIN (6)
^FORM/SHAPE/MARGIN (7)
>___________________medusa head/ lion face
>___________________swarming colonies
>___________________rough edges
>___________________star colonies with feet
>___________________serrated colonies
>___________________molar tooth colonies

^__________________-Determined by tilting the plate and looking at the side of the colony: (6)
>________________________- umbilicate
>________________________-umbonate
>________________________-convex
>________________________-generally flat

OPTICAL PROPERTY: (3)

>___________________- If colonies allow light to pass through
>___________________- if colonies allow some light to pass through (E.g: 1)
>___________________- if colonies do not allow light to pass through (E.g: 1)
*_____________________- shiny, half pearl on BAP
HEMOLYTIC PATTERN (3) *(2)
>_______________________- incomplete hemolysis; greenish zone around colonies
>_______________________-complete hemolysis; clear zone around colonies
>_______________________-non hemolytic; no zone around colonies
*_______________________-Small zone of alpha hemolysis and beta hemolysis surrounds the
colonies usually after refrigeration
*_______________________-characterized by beta hemolytic inner zone and alpha hemolytic
outer zone (e.g 1)

SIZE OF COLONIES
-can be described as large, medium and small
^______________________- pinpoint colonies
^______________________-pinhead colonies

CONSISTENCY AND TEXTURE: (4)

-Determined by touching the colony with a loop
^__________________- crumbly like bread crumbs and wrinkled (E.g. 1)
^__________________-butyrous like butter (E.g.1)
^__________________- (E.g 2)
^__________________- (E.g. 1)

COLOR: can be white, gray, yellow of buff: (4)

PIGMENT: inherent characteristic of specific organism confined generally to the colony (5)
^_____________________________________-blue green
^_____________________________________-red
^_____________________________________-blue
^_____________________________________-violet
^_____________________________________-brown black
ODOR: Determined when the lid of the culture plate is removed (5)
^________________________-Old rock smell
^________________________- Fruity or grape like
^________________________-Putrid odor
^________________________-Musty basement “mousy”
^________________________-freshly plowed field scent

GROWTH IN BROTH/SEMI SOLID MEDIA: (6)

^________________________-vine or streamer effect
^________________________-Puffed balls
^________________________-Scum at the sides of tube
^________________________-Umbrella shape in semi-solid medium
^________________________-Test tube brush growth in semi-solid medium
^________________________-stalactite pattern in broth

ANTIMICROBIAL SUSCEPTIBILITY TESTING:

^___________________________-used to determine the bactericidal level of antibiotic in a
patient’s serum against the organism isolated from the patient. AST is usually done in-vitro
method
>___________________________-Quantitative test in which the serial dilution of antibiotics
are prepared and standard concentration of bacteria is added
 TSIA IVMC LOA PD PD H2S Ureas Urease ONPG ONPG ST PCM LDP ADN MALT S/M MOT IN
(-) e (-) (-) (-)
E.coli