lab 1

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Which 2 main genres form endospores?

Bacillus & Clostridium

Gamma-hemolytic

Bacteria that cause no change in blood agar

What is the reagent in the VP test?

Barretts Reagent A Barretts Reagent B

Schistosoma

Blood Fluke

Proglottids

Body segments of a worm

Clostridium Botulinum

Botulism

Nostoc

Bright Blue Curly "q's"

which endospore forming bacteria causes deadly diseases?

C. perfringens , B. cereus

Iodine

Chemical that causes starch to turn brown-black if present in Amylase test

Spore-forming species

Clostridium & Bacillus

Endo Agar

Coliforms produce a gold metallic sheen in which test medium?

Yellow

Color Mannitol/Salt agar turns when mannitol is fermented by the bacteria on it

What is the best way to visualize capsules?

Combine a negative stain and a simple stain

What occurs in β-hemolysis?

Complete destruction of RBCs and hemoglobin

Diatoms

Cool Shapes

Primary stain

Crystal Violet/Gram +/1 min

a key for the identification of organisms based on a series of choices between alternative characteristics

Define Dichotomous key

DNase agar - enzyme?

Deoxyribonuclease

Citrate Utilization Test

Differentiates the ability to ferment citrate as a sole carbon source (+) blue -- E. aerogenes (-) green -- E. coli

No

Does staphylococcus epidermidis cause hemolysis?

Casein Hydrolysis

Does this organism produce the enzyme casease?

Gas tubes in some phenol red tests that collect a bubble of gas if bacteria in them produce gas upon fermentation of carbohydrates

Durham gas tubes

What organism tends to produce a green metallic sheen on EMB?

E. coli

EMB plate select for what group of bacteria

E.coli antibacterial

**Selects for gram - ***Differential for lactose, lactose + = red colonies and surrounding medium **Coliforms produce a gold metallic sheen

Endo Agar test

True/False: Aspergillus flavus causes ergot poisoning.

False

Station 16 Malaria Vector

Female Anopheles mosquito

What is the hydrogen sulfide indicator in TSIA?

Ferrous sulfate

Contaminated inanimate object

Fomite

"Microscopic Inversion"

From what is seen with the naked eye, whatever is being viewed under Brightfield Microscope is rotated 180 degrees and moves left when shifting right.

Neisseria

GN diplococcus

Hemophilus

GN pleomorphic rod

Enterobacter

GNR

Escherichia coli

GNR

Klebsiella pneumonia

GNR

what does a mordant do?

Helps lock in the dye to the cell wall

Ancylostoma

Hookworm

10 years with no symptoms

How long does the incubation period of HIV last sometimes?

Ascaris

Human Roundworm

What reagent is used for a catalase test?

Hydrogen peroxide

what does heat-fixing do?

"fixes" the bacteria to the slide so they do not move and can retain their original shape

In lab 9, what was the name of the person who originally had the disease?

"patient zero"

Triple Sugar Iron (TSI) test, Hydrogen sulfide, yellow = +

(agar inside tubes) What test? What causes one tube to turn black? What color is positive?

(Antimicrobial agent) Inhibition of nucleic acid synthesis EXAMPLE

**Cipro [Ciprofloxacin]

Result of Gram Stain

-Staph epid (gram+ cocci) are stained purple from the Crystal Violet (due to thick cell walls) -E. coli (gram- rods) are stained red from the Safranin

BA - gamma hemolysis

-no hemolysis - turn into white colonies

Mycotoxicoses

-occur following consumption of food that has been poisoned by fungal toxins (mycotoxins) -examples: Aspergillus flavus & Claviceps purpurea

Motility Test - motile negative?

-organism will only grow along the streak line -medium will NOT be cloudy

Darkfield Microscope

-specimens appear bright against a dark background -observes LIVE specimens -doesn't require staining

What two items did we use in lab to make E.coli competent?

-transforming solution (CaCl2) -heat shock

Microsporum (Tinea capitis)

-type of mycoses = cutaneous -phylum = Deuteromycota -entry route = direct contact (skin) -treatment - Nystatin -disease = Ringworm; Athlete's foot

Sporothrix schenckii (Fungus) disease = Sporotrichosis

-type of mycoses = subcutaneous -phylum = Deuteromycota -entry route = direct contact (skin) -occupational hazard for gardeners -treatment = Amphotericin B and/or potassium iodide

Motility Test - procedure?

-use a needle to inoculate by making a single stab 2/3 down -incubate at optimum temp for 24-48 hrs

Amount plated will always be how many milliliters?

.1 or 10^-1

Factors that determine the efficiency of disinfectants and antiseptics

1. Concentration 2. Length of Exposure 3. Resistance 4. Temperature, pH, type of material

Name the steps of a Gram stain

1. Crystal violet (primary stain) 2. Gram's iodine (mordant) 3. ethanol (decolorizing agent) 4. safranin (counter stain)

What 3 bacterial activities does a SIM medium detect?

1. Sulfur reduction 2. Indole production from tryptophan 3. Motility

Making a Standard Smear

1. Tiny loop full of water on slide 2. Use sterile (heat) inoculation loop to transfer TINY amount of microorganism into water and spread into bigger circle. 3. Air dry slide COMPLETELY 4. Heat fix

How many WBCs are counted and tallied in a differential white cell count?

100

What total magnification is the Oil Immersion objective? What color is the ring?

1000X/Black

What total magnification is the low power objective? What is the color of the ring?

100X/Yellow

What is the diameter of all the granulocytes?

12-15 micrometers

What is the diameter of monocytes?

12-20 micrometers

What reagent is used for DNAse test?

1N HCl

How many lobes does the eosinophils have?

2

Binocular

2 occulars for depth perception

Acetoin & 2,3 butandiol

2 products detected in the Voges-Proskaeur Test

Normal % of lymphocytes?

25-33%

Dextrose (glucose), lactose, sucrose

3 carboydrates that produce enzymes during fermentation and can be tested in the lab

Pseudomonas aeruginosa, Escherichia coli, Staph. (aureus & epidermidis)

3 catalase positive bacteria

objective lenses

40x, 10x, 40x, 100x

URINE figure a colonie count?

50 colonies = 50,000 greater than 100,000 to be treated (colonies x 100,000)

Normal % of neutrophils?

55-65%

Phenol Red is pink above what pH?

7.4

Phenol red is red at what pH range?

7.4 - 8.4

E TV SLUMS 1. EMB (Eosin Methylene Blue) 2. TSI 3. VP 4. Sucrose 5. Lactose 6. Urease 7. Methyl Red 8. Simmons Citrate

8 Tests commonly run on Gram negative bacteria

Phenol red is pink above what pH?

8.5

What's the Rh factor?

A clumping factor in the blood.

Mycolic wax

Acid-fast microorganisms have a high ___________content in their cell walls, which resists staining.

Reason for extended heat in Acid-Fast Stain

Acid-fast species have a waxy cell wall, therefore color must be melted into the cell wall.

What are the color changes for neutral red?

Acid=hot pink Neutral=dusty rose Alkaline=tan

Bacillus Anthracis

Anthrax

Indirect ELISA test for...

Antibodies

The Kirby-Bauer test measures:

The effectiveness of antimicrobics against pathogenic microorganisms

In a Kirby-Bauer test, the zone of inhibition depends on:

The sensitivity of the bacteria to the specific antimicrobial agent

Epidemiology

The study of causes, occurrence, transmission, distribution, and prevention of disease.

What do ureases do?

They cleave urea into ammonia and carbon dioxide, causing an increase in pH

Hydrogen sulfide

This chemical is present if TSI agar turns black

??

This parasite only occurs in the United States and is the chief cause of morbid obesity.

E. coli and Proteus

This person has a UTI. What bacteria is causing it?

A+

This person has what type of blood?

True/False: Infected Anopheles mosquitoes can transmit malaria.

True

False

True/False: The "scotch tape test" is used to detect the presence of hookworms.

Station 5 Urease test Substrate

Urea

Station 5 Urease test Enzyme

Urease

Why did we use an index card or leave the lid on during the experiment?

We wanted to see growth of bacteria without UV light interaction

ELISA, Western blot as a confirmatory test

What are the 2 methods used to diagnose HIV?

BLUE= O2 present WHITE= No O2 present

What do the colors in the anaerobic jar indicate?

Facultative anaerobe Growth with or without O2 Growth throughout test tube.

What organism does the broth show?

Detect the antibodies produced by our body to fight HIV.

What part of the virus does the Elisa test detect?

Positive: Staph Aureus Negative: Staph epidermis

What will the results of the Catalase test indicate?

Mutualism

When an organism benefits from us while giving us something in return

Parasitism

When an organism benefits from us while harming us

Commensalism

When an organism benefits from us without giving us anything in return

What is transformation?

When bacteria can pick up DNA from the environment and make use of the genes it carries

Streptococcus pneumoniae

Which bacteria is sensitive to Optichin or P disc antibiotic?

E. coli & Enterobacter

Which bacteria will grow on EMB?

Shigella Blue green color with no black precipitate due to NO H2S production.

Which enteric microorganism could be on this Hektoen media and why?

reproductive spores can become airborne leading to laboratory acquired respiratory infections

Why is it dangerous to work with pathogenic fungi in the laboratory?

How fast should a positive result appear for an oxidase test?

Within 20 seconds

Pandemics

Worldwide epidemics

Mannitol fermenter results in what color?

YELLOW

Station 2 Phenol Red media Negative result:

Yellow - has used carbohydrate, produced acid

What method do we use for the acid fast stain?

Ziehl-Neelsen

Where growth is inhibited

Zone of inhibition

What do Gram +'s have that Gram -'s don't

a thick cell wall that contains peptidoglycan and teichoic acid that holds in the crystal violet

A disease such as AIDS will make a patient susceptible to a respiratory disease called: a. PCP b. jock itch c. ringworm d. ergot

a. *PCP* b. jock itch c. ringworm d. ergot

Voges-Proskauer "MR-VP Broth" - purpose?

ability of an organism to produce acetoin, 2,3 butanediol

Methyl Red Test "MR-VP Broth" - purpose?

ability of an organism to produce mixed acid fermentation

Urea Hydrolysis "Urea Broth" - purpose?

ability of an organism to split urea to form ammonia "alkaline" by the action of urease -medium contains phenol red - turn pink in alkaline conditions

Resolution

ability of lens to distinguish between objects close together expressed as a distance

Nitrate Reductase Test (Nitrate Broth) - purpose?

ability of organism to reduce nitrate (NO3) to nitrite (NO2) or N2 by production of nitratase

define competency

ability to take and express recombinant DNA

Citrate Utilization - positive result?

alkaline pH makes it change from green to Prussian blue

What are obligate aerobes?

bacteria that require oxygen to grow, so they grow at the top of the tube and often the methylene blue is gone.

Hemophilis influenza

bacterial meningitis

Crystal Violet - primary dye

basic and carries positive charge. interacts with negatively charged cell wall

H2S+ =

black precipatiate

Microscope eyepiece

contains a 10x lens

URINE if plate contains multiple organisms you would report?

contamination

Rheostat?

controls light density, less light intensity will be needed with low power objectives

What does Hemolysin do?

destroys rbc's

Fluid Thioglycollate Medium "Oxygen Requirements" What does this medium used for?

determiine oxygen requirement of medium

Motility Test - purpose?

determine whether or not an organism is motile

red blood cell

disc shaped

Gelatinases hydrolyze

gelatin

Neisseria gonorrhea

gonorrhea

how long can endospores last?

indefinitely

antibodies used the diagnostic kits are made how

injecting rabbits with the antigen

EMB agar - differential agents

lactose and sucrose

Agranulocytes

lymphocytes monocytes

BA - complete hemolysis "beta"

lyses cell membranes of all RBC's

colony forming units (CFU) =

number of colonies counted divided by the amount plated in ml times the dilution (# colonies counted/(amt plated x dilution))

What makes DNA negative?

phosphate group (PO4-)

What does catalase do?

protects a cell against hydrogen peroxide by breaking it down into water and oxygen

gram positive rod

purple

What color do eosinophils stain?

red

gram negative cocci

red round shaped

What is Counterstain

safranin/1 min

HE agar selects of bacteria

seracisous marcesci

define inoculum

small number of cells added to a sterile medium to create a pure culture

what does the media in citrate utilization contain

sodium citrate- carbon source. ammonium dihydrogen phosphate- nitrogen source. bromothymol blue- pH indicator.

if an organism is a non-ferreter what would you see in a macconkey agar part

tan

Direct ELISA - what are we testing for?

test for antigens "pathogen"

On what basis are the protozoa classified?

the 4 classes of protozoans are based on their type of motility

What is minimum inhibatory concentration (MIC)?

the exact value of the smallest amount of antibiotic that kills/inhibits the microorganisms

What does the EMB medium have?

the medium has two indicators dyes eosin and methylene blue.

Oxidase Test - purpose?

to see if an organism is an aerobe -it checks for the ETC

What is the total dilution of the following dilutions: 1/100 X 1/10 X 1/10 X 1/10 X 1/10 X 1/10 X 1/10

total dilution = 1/100,000,000

What is a positive test in the ELISA test look like?

turn a blue color

Salmonella typhi

typhoid fever

proteus mirabilis changes broth to bright pink, this is a positive result.

urea broth

Most effective antibiotic:

varies... see results chart

Least effective antibiotic:

varies...see results chart

Aerial hyphae goes

vertically

Results of LB/AMP/ARA with + pGLO?

very active growth and will glow

Do bacteria have the ability to produce endospores?

yes long gram positive rods produce endospores

Since agar is not degraded by most bacteria, can it be liquified

yes, at 100 degree Celsius. it will remain a liquid at temperatures above 40 degree Celsius, allowing growth media to be poured into petri plates or tubes.

Test #2-Indole Test

•AKA tryptone •indicates the ability of bacteria to conver tryptophane → indole •KOVACS REAGENT is addded to detect indole → red lining •All ESCHERICHIA COLI , by definition, are indole positive.

What shape is spirillum/Spirilla (pleural)

"S" shaped

(Antimicrobial agent) Disruption of cell membrane function EXAMPLE

**Anti-fungal Drugs (such as ClotrimAZOLE, MiconAZOLE, KetaconAZOLE)

(Antimicrobial agent) Inhibition of cell wall synthesis EXAMPLE

**Penicillin (or Cephalosporins, Vancomycin, INH)

(Antimicrobial agent) Interference with enzyme function EXAMPLE

**Sulfa Drugs [Sulfonamides]

(Antimicrobial agent) Inhibition of protein synthesis EXAMPLE

**Tetracyclines (such as Minocycline & Doxycycline)

Staphyloslide Latex Agglutination Test

**wear gloves -when Staph aureus' surface proteins bind with human IgG antibodies, results in agglutination -Staph aureus is the ONLY positive reaction -Positive: speckled, agglutinated dots -Negative: no/almost no speckled dots

ELISA test in lab - 5 steps.

*3 drops of antigen *3 drops of patient's serum and also a +/- control *3 drops of antibodies linked with enzyme *3 drops of chromagen *wait 5 minutes for results

3 differential stains

*Acid Fast *Spore Stain *Gram Stain

A red color after reagents in a nitrate test indicates:

+ Nitrate reduction to nitrite

No color change after reagents and zinc in a nitrate test indicates:

+ Nitrate reduction to nongaseous nitrogenous compounds

Tapeworm Scolex

-"head end" of tapeworm -note hooks and suckers -member of Platyhelminthes -used to attach to intestinal wall

Cestodes (tapeworms)

-20-30 feet in length -most are hermaphroditic (this allows for self-fertilization) -characterized by having a head end (scolex) which contains hooks and suckers used for attachment to intestinal wall and a body that's divided into segments (proglottids) -these units contain the reproductive and digestive systems of the worm -the scolex MUST be removed to ensure elimination of worms since as long as it remains attached, proglittids will continually be formed and released

Aspergillus Species (mold)

-A. niger a common black mold (causes "moldy" bread) -A. fumigates can cause respiratory problems (aspergillosis) -A. flavis can produce a poisonous aflatoxin -Aflatoxin may contaminate peanut butter, peanuts, rice, grains -toxin in grains responsible for large wild bird kills -supportive treatment -inhalation of spores or consumption of toxin if food *note the "starburst" conidiophore*

Kirby-Bauer/"Disk Diffusion Method"

-Accounts for many variables that may impact test results -Mueller-Hinton agar -4mm thick -pre-soaked absorbent paper disks -agar thoroughly covered in microorganism with cotton swab -16 to 18 hours incubation -measure zones of inhibition in millimeters and compare to specific diameters in standardized table

What allows the test for ELISA to be (+)?

-Antigen is in the well...when patient has the antibody it will bind with the antigen. -once the antibody linked with enzyme enters, this will attach to the patients antibody. -when chromagen enters it will react with the enzyme that is attached the the complex and show a blue color

Oxygen Requirements "Fluid Thioglycollate Medium"

-Areobes -growth on top-pseudomonas aerigenosa -Facultative-growth throught the medium-Staphylococcus aureus & E. coli -Anaerobes-growth on bottom-Clostridium

ONPG Test

-B-galactosidase breaks down lactose.

Results of Blood Agar Hemolytic Types

-Beta: RBC completely destroyed -Alpha: RBC partially destroyed -Gamma: no RBC destruction

Types of Hemolysis (Results of Blood Agar)

-Beta: clear (maybe yellow) zone around colonies... b/c RBCs completely destroyed -Alpha: greenish-gray zones... b/c RBCs partially destroyed -Gamma: really no change... b/c no RBC destruction

Sterile Specimen Collections

-Blood & other [internal] body fluids (CSF, synovial, pleural, ect.) -Urine: before voiding and if clean catch, midstream -[Deep] Wound specimen: not always sterile but normal flora are not expected

Results of Citrate Test

-Blue: Citrate positive -Green: Citrate negative **if there is any blue on a Citrate agar plate = Citrate positive

Oxygen Requirements "Fluid Thioglycollate Medium"

-Boil 5 min -loose cap -tighten cap -Allow to cool -donot shake incubate 24hrs

Match the items listed in the "KEY" below to answer questions KEY: a. Candida b. Balantidium c. Cryptococcus d. Schistosoma e. Wucheria -Can be acquired from breathing contaminated bird feces -Can affect little children and cause diaper rash -Can cause swelling of body parts

-Can be acquired from breathing contaminated bird feces = C -Can affect little children and cause diaper rash = A -Can cause swelling of body parts = E

Match the items listed in the "KEY" below to answer questions KEY: a. Sporotrichosis b. Bilharzia c. Schistosomiasis d. Pinworms e. Trichinosis -Causes intense anal itching. -Is caused by a fungus which is an occupational hazard for gardeners. -The larvae burrow through your skin when you wade in contaminated water. -May cause "swimmers itch". -Acquired from eating contaminated pork and other meats.

-Causes intense anal itching = D -Is caused by a fungus which is an occupational hazard for gardeners = A -The larvae burrow through your skin when you wade in contaminated water = B -May cause "swimmers itch" = C -Acquired from eating contaminated pork and other meats = E

Ways to improve resolution on Brightfield Microscope

-Condenser lens as high as possible -Diaphragm opened to shine as much light as possible -Use immersion oil to reduce scattering of light

Trypanosoma brucei, gambiense, and rhodesiense (Mastigophora-Flagellate)

-DISEASE = African sleeping sickness -GENERAL SYMPTOMS = severe anemia leading to fatigue, weakness, coma, and death; neural; circulatory -TRANSMISSION = Tsetse fly bite; cattle and wild animals represent nartural reservoirs -TREATMENT/PREVENTION = avoid fly bites; Suramin and melarsoprol

Food Microbiology - Serial Dilution and Plate Counting

-Dilution Tubes - 9 ml saline tubes -Dilutions made with 1 ml pipettes -Plate Counting: 30-300 colonies -Quebec Colony Counters used -formula: Milk: TVB/ml = PC X DF Meat: TVB/gm = PC X DF

Which of the parasites studied commonly occur in the United States?

-E. histolyticum -B. coli -Giardia -Trichomonas -Trypanosoma cruzi -Toxoplasma gondii -Cryptosporidium; Plasmodia

Balantidium coli (Ciliate) DISEASE = Balantidasis or balantidial dysentery

-GENERAL SYMPTOMS = intestinal infection; may cause ulceration of GI tract -TRANSMISSION = contaminated food/water; poor hygiene -TREATMENT/PREVENTION = Metronidazole; Chlortetracycline -forms both trophozoites and cysts -normal intestinal inhabitant in swine and frogs -the only ciliate to cause disease *very large, round to oval parasites w/macronuclei*

Trichomonas vaginalis (Mastigophora-Flagellate) DISEASE = Vaginitis; UTI

-GENERAL SYMPTOMS = itching; pain; foul discharge -TRANSMISSION = sexual contact -TREATMENT/PREVENTION = abstinence; Metronidazole -opportunistic parasite that can invade when acidity of urinary tract changes *look for large oval flagellated cells*

Endamoeba histolyticum (Amoeba) DISEASE = Amebiasis or amoebic dysentery

-GENERAL SYMPTOMS = severe intestinal disease; ulceration and perforation of GI tract -TRANSMISSION = contaminated food/water; poor sanitation; filth -TREATMENT/PREVENTION = Metronidazole *look for oval cells with nuclei* *see both cysts and trophozoites*

Leishmania donovani & tropica (Mastigophora-Flagellate) -DISEASE = Leishmaniasis -visceral disorder called Kala-azar (caused by donovani); skin sores (tropica causes Oriental sores)

-GENERAL SYMPTOMS = systemic -TRANSMISSION = Phlebotomus sand flies -TREATMENT/PREVENTION = avoid sand fly bites; chemo; surgery -common in Asia, Africa, & Middle East -tissue section; amastigote form *look for very tiny Donovan's bodies*

Psuedomonas aeroginosa

-Gram (-) rod -Green pigment -Fruity odor -Resistant to many antibiotic; only sensitive to Cipro -Common in ICU, trauma, and burn units

Match the items listed in the "KEY" below to answer questions KEY: a. Protozoan b. Tapeworm c. Fungus d. Fluke -Histoplasma capsulatum -Fasciola hepatica -Giardia intestinalis -Taenia solium

-Histoplasma capsulatum = C -Fasciola hepatica = D -Giardia intestinalis = A -Taenia solium = B

Result of Capsule Stain

-India Ink (- charge) stains the background a dark [black-ish] color -Crystal Violet (+ charge) stains Bacillus Megaterium blue

Antimicrobial Agents: Mechanisms of Action

-Inhibition of cell wall synthesis -Inhibition of protein synthesis -Disruption of cell membrane function -Inhibition of nucleic acid synthesis -Interference with enzyme function

Slide agglutination test

-Known antibodies are mixed with the organism in question -Positive: speckled, agglutinated dots -Negative: no/almost no speckled dots

Positive bacteria for Snyder test

-Lactobacilli -Oral Streptococcus mutans

Result of Endospore Stain

-Malachite Green stains Bacillus Megaterium and its endospore green then water decolorizes this water soluble stain out of the bacteria but leaves the spore green -Safranin stains the bacteria red

Nucleic Acid Test

-Patient DNA/RNA compared to known DNA/RNA of organism in question -Ex: Gonorrhea, Syphilis, etc.

Results of MacConkey Agar

-Pink: positive; ferments lactose -Flesh-like/no color: negative; does not ferment lactose

Classification of Helminthes

-Platyhelminthes (flatworms) -Nemathelminthes (roundworms) -each is further divided into classes: 1. tapeworms = Cestoda 2. flukes = Trematoda 3. roundworms = Nematoda

Match the disease to the appropriate method of treatment/prevention/diagnosis: a. wind up on a stick b. control "no see um" sand flies c. use antifungal ointment d. use scotch tape e. hope for the best -Ringworm infections -Pinworm infections -Oriental sores -Guinea worm infections -Naegleria infections

-Ringworm infections = C -Pinworm infections = D -Oriental sores = B -Guinea worm infections = A -Naegleria infections = E

Result of Acid-Fast Stain

-Staph epid (non acid-fast, gram+ cocci) are stained blue from the Methylene Blue (first stained red by Carbofuchin then decolorized by the acid alcohol and stained with Meth Blue) -Mybacterium (acid-fast rods) are stained red from the Carbofuchsin

MSA - example bacteria

-Staphylococcus aureus - yellow "low pH" - positive for fermentation - Staphylococcus epidermis - pink "high pH" -negative for fermentation

Normal Flora Present Specimen Collections

-Throat swab -Sputum -Fecal (stool) -Vaginal

Antibody-Based Quick Strep Test

-Top line will always show, meaning test worked -2nd line shows, meaning Positive reaction

Catalase Test

-Used along w/other Gram (+) tests -Used to tell between Staph (catalase +) and Strep (catalase -) -Converts Hydrogen Peroxide to water and oxygen

Coagulase Test

-Used to specify types of Staph -Staph aureus is the ONLY positive reaction; all other Staph are negative -Positive: clumps up (clots) -Negative: doesn't clump (clot) -Uses Rabbit Plasma (contains fibrin: clotting factor)

Results of Mannitol Salt Agar Test

-Yellow: positive; ferments mannitol -Pink: negative; does not ferment mannitol -Growth: salt tolerant -No growth: not salt tolerant

ELISA direct test

-add patient serum (if + it will have Ag) -add antibody with linked enzyme -add chromagen -wait for color change

Helminth Reproduction/Life Cycles

-adult parasites may exhibit separate sexes or have both combined w/in one animal (hermaphroditism) -production of thousands of eggs (ova) which may be released in stool of host -man may harbor adult parasites or some other form including the larvae -some utilize intermediate hosts such as snails or fish to maintain life cycle

Toxoplasma gondii (Apicomplexa) DISEASE = Toxoplasmosis

-affects any organ of body especially brain, heart, and lymphatics; congenital defects -TRANSMISSION = infected cats; cat feces; under-cooked meat -Sulfonimides -affects immune compromised, fetus of pregnant women *look for crescent-shaped parasites*

Protozoan Structure

-all protozoa possess a cell membrane or a pellicle and one or more nuclei w/in cytoplasm -obtain food by ingestion, diffusion thru cell membrane, pinocytosis, or phagocytosis (amoebas)

What is the primary difference between arachnids and the insects?

-arachnids possess 4 pairs of legs, 2 mouth parts, no antennae -insects possess 3 body parts, 6 legs, antennae, and mandibles or jaws

Treatment/Prevention of Mycoses

-avoiding inhalation of asexual spores found in dust -treatment depends on the body area or tissue involved -topical ointments: tinactin and griseofulvin used for CUTANEOUS -amphotericin B, nystatin, and the azoles (ketoconazole) for SYSTEMIC

Nitrate Reductase Test - procedure?

-boil 5 minutes -tighten cap and let cool -inoculate broth -incubate at optimum temp -add 5 drops each of Nitrate Reagent A and B

Citrate Utilization - contents?

-carbon source -sodium citrate -nitrogen source -ammonium salts -pH indicator -brom thymol blue

Disease: Cysticercosis

-causative agent = Taenia solium -mode of transmission = ingestion of pork containing tapeworm eggs - treatment = none

Naegleria fowleri (Amoeba)

-causes amoebic meningo-encephalitis -naturally in brackish, muddy water of ponds and lakes and invade sinuses of normal healthy children

Trypanosoma cruzi (Mastigophora-flagellate) DISEASE = Chagas disease

-causes ulcers; cardiovascular; bite and then fecal deposit of Triatomid/Conenose bug -treatment/prevention = avoid bug bites; reservoirs seem to be small rodents, opossums, & armadillos; endemic in Mexico and S and Central America (myocarditis) *look for flagellated crescent-shaped parasites in blood smear*

Why are stool samples concentrated to demonstrate intestinal parasites?

-concentration increases the chances of finding the parasites -some stool samples may only contain a few making it difficult to identify

Cultivation and Dx of Mycoses

-cutaneous agents = wet mounts w/KOH or lactophenol cotton blue; fluorescent stains; tissue stains -systemic infections = lung aspirates; tissue biopsies; stains and X-rays -cultivation = Sabourauds dextrose agar and mycosel agar

Protozoan Cultivation and Dx

-depends on type of disease but variety of clinical samples (stool, blood, spinal, vaginal secretions) may be needed -various staining techniques used to observe them (hematoxylin or trichrome) -diagnosis following O & P (ova and parasite) check usually confirmed by observation of cysts or trophozoites in concentrated sample

Scanning Electron Microscope

-detailed 3D view of specimen's surface

EMB agar - purpose of differential agents?

-differentiate coliform -color change for fermentation

Tsetse Fly (Vector = Glossina Fly)

-disease association = African sleeping sickness (protozoa) -mechanical (M) or biological (B) = B -initial source of parasite = wild animals; cattle -arachnid or insecta = insecta -Suramin = treatment -transmits trypanosomes through bite -only occurs in Africa -causative agent = Trypansoma brucei gambiense & T. brucei rhodesiense *look for flagellated crescent-shaped parasites in blood*

Common House Fly (Vector = Musca & Sarcophaga)

-disease association = Amoebic dysentery (protozian cysts) and pinkeye (bacteria) -mechanical (M) or biological (B) = M (mechanically transmit anything -initial source of parasite = anywhere -arachnid or insecta = insecta *PEST*

Rat Fleas (Vector = Xenopsylla cheopis)

-disease association = Bubonic or Black Plague (bacteria); murine typhus (rickettsia) -mechanical (M) or biological (B) = B (may mechanicaly transmit tapeworms) -initial source of parasite = rats; rodents -arachnid or insecta = insecta (bloodsucking ectoparasites) -causes fever; rash -possess incredible jumping ability

Conenose bug or kissing bug (Vector = Triatomine bug)

-disease association = Chagas disease -mechanical (M) or biological (B) = B -initial source of parasite = rodents; unsanitary conditions -arachnid or insecta = insecta -transmits trypanosomes through bite and fecal deposit of parasites -found in Mexico, Central & South America & in TX -active @ night and inflicts painful bite

"Crab" or Pubic Lice (Vector = Phthirus Lice)

-disease association = Crabs -mechanical (M) or biological (B) = M -initial source of parasite = humans -arachnid or insecta = insecta -causes itchy infestation of skin, transmitted by close personal and sexual contact -eggs called nits and stick tightly to hairs of vicitm; treat w/Kwell

Mosquito (Vector = Aedes Mosquito)

-disease association = Dengue fever, Yellow Fever, & West Nile fever (transmits viruses) -mechanical (M) or biological (B) = B -initial source of parasite = animals; humans -arachnid or insecta = insecta -causes high fever; chills; pain

Body Lice (Vector = Pediculus Lice)

-disease association = Epidemic typhus (rickettsial); Relapsing fever (bacterial) -mechanical (M) or biological (B) = B -initial source of parasite = humans -arachnid or insecta = insecta -causes severe rash/fever -hide in clothing and have mouth parts adapted for sucking blood/tissue

Mosquito (Vector = Anopheles Mosquito)

-disease association = Malaria & elephantiasis (protozoa) -mechanical (M) or biological (B) = B -initial source of parasite = animals; humans -arachnid or insecta = insecta -causes high fever; chills

Hard Ticks (Vector = Dermacentor & Amblyomma ticks)

-disease association = Rocky Mountain Spotted Fever (rickettsia), Lyme Disease & Tularemia (bacteria); or viruses (encephalitis) -mechanical (M) or biological (B) = B -initial source of parasite = wild animals (small animals deer) -arachnid or insecta = arachnida -causes rash; fever; pain (infect man while feeding on blood) -transovarian transmission may occur

Scabies (vector = Sarcoptes mite)

-disease association = Scabies -mechanical (M) or biological (B) = M -initial source of parasite = rodents; animals -arachnid or insecta = arachnida -infests by burrowing into skin; highly contagious and cause intense itching -scratching aggravates condition -treat w/gamma benzene hexachloride (Kwell)

Cockroaches (Vectors = Blaberus & Blatta)

-disease association = none -mechanical (M) or biological (B) = M -initial source of parasite = unsanitary conditions -arachnid or insecta = insecta -Blatta acts as M vector for bacteria (salmonellois) and protozoa (amoebic dysentery); Blaberus acts as M vector for bacteria -primitive mouth parts and can ONLY be M vectors *PEST*

Sand Flies (Vector = Phlebotomus Flies)

-disease transmitted = Leishmaniasis (protozoa) -mechanical vector -causes oriental sores; systemic

Deer Flies (Vector = Chrysops)

-disease transmitted = Tularemia (bacteria), Loa Loa (filarial worms) -biological vector -causes Bobos; eye worm

Screwworm Flies (Vector = Cochliomyia Flies)

-disease transmitted = larvae cause worm myiasis -mechanical vector -causes flesh eating maggots (freely moving larvae)

Sand Fleas (Vector = Tunga Fleas)

-disease transmitted = none -mechanical vector -pest; produce intense itching leading to ulceration

Bedbugs (Vector = Cimex bugs)

-disease transmitted = none -mechanical vector carrying bacteria -pest

Soft Ticks (Vector = Ornithodorus tick)

-disease transmitted = rickettsia (tick paralysis) or bacteria (relapsing fever) -biological vector -infect man while feeding on blood

Trichomonas tenax (Mastigophora-Flagellate)

-disease= Gingivitis -symptoms= oral discomfort -transmission= kissing? -prevention= Good oral hygiene

Life Cycle of a Cestode

-distal, mature proglottids, which are filled w/eggs, may be shed in feces of host and if not properly disposed, could contaminate the soil or vegetation -the egg filled proglottids may be picked up by another unsuspecting host and cycle is complete

Why do we use bacteriostatic dyes in culture media? List two examples of media containing dyes.

-dyes provide a means of selection -dyes such as methylene blue and crystal violet inhibit G+ bacteria -examples: EMB, MAC

Life Cycle of a Trematode

-eggs produced by adult worms are eliminated in feces of infected animal -water access = eggs hatch into larvae called miracidia (infect aquatic intermediates -snails; crayfish) -change into infective larvae called cercaria; reenter new host by burrowing thru skin/ingestion of contaminated vegetation -larvae mature in blood stream into adults in host organs (liver, lungs, bladder) -after fertilization, female migrates to intestinal tract-releases eggs

Why are bacterial endospore monitoring devices more effective in evaluating sterility than heat sensitive tape?

-endospore devices react to pressure, temperature, and time -heat sensitive tape only reacts to temperature not pressure or time -therefore, the tape does not indicate sterility

Pneumocystis carinii (Opportunistic Fungus) disease = PCP; Pneumocystic pneumonia

-entry route = nasal -aerosols; compromised defense systems -common cause of death for patients with AIDS -Pentamidine and trimethoprim w/sulfa-methoxazole *lung smear of patient with PCP* *look for oval cysts in lung tissue*

Aspergillus Flavis (Mold)

-food poisoning (infected food entry) -Mycotoxicosis (produces carcinogenic mycotoxin called aflatoxin) -phylum =Ascomycota -elevated levels of aflatoxin in peanut butter, peanuts, rice, and cereal grains result in recalls *notice "starburst" conidiophore*

Cutaneous Mycoses (Dermatophytoses)

-fungi capable of degrading keratin and affecting skin, hair, or nails -infections commonly called tinea or ringworm since lesions tend to be circular -most infections respond to topical treatment, however oral griseofulvin may be used -transmitted directly via animal/human carriers thru infected hair or skin -example: Tinea capitis

Plasmodium malariae, falciparum, & vivax (Apicomplexa) DISEASE = Malaria

-general symptoms = fever; chills; weakness; vomiting -transmission= female Anopheles mosquito; treatment = Quinine derivatives -falciparum species highly resistant and often fatal species *look for rings/half moons in blood cells*

Endamoeba gingivalis (Amoeba) DISEASE = Oral Gingivitis

-general symptoms= oral discomfort -transmission = kissing -treatment/prevention = good oral hygiene

Systemic Mycoses

-generally begin as respiratory infections; but capable of dissemination to skin or other organs -inhalation of spores probable route of transmission -spores DO NOT seem to be contagious either from infected animals or humans -treatment = Amphotericin B -Cryptococcus (Filobasidiella) neoformans; Histoplasma capsulatum; & Coccidioides immitis

BA - incomplete hemolysis (alpha)

-greenish tinge -RBC's lyse midway -hemoglobin is reduced to methhemoglobin which is release through bilirubin

*Intestinal Nematodes*: Necator americanus & Ancylostoma duodenale (adult hookworms) *intestinal nematodes - note mouth parts and hooks*

-humans; dogs (penetration of infectious larvae while walking barefoot thru fecal contaminated soil) larvae migrate around tissue (cause cutaneous larva migrans); gain access to blood stream-to heart, lungs; coughed up, swallowed, & into intestines -abdominal pain; bloody diarrhea, anemia; Mebendazole & Ivermectin

Gelatinase - what can it do?

-hydrolyze gelatin into polypeptides -hydrolyze gelatin polypeptides into amino acids

Phase Contrast Microscope

-improved visual of internal details -observes LIVE specimens -doesn't require staining

Carbohydrate Fermentation Test - procedure?

-inoculate each medium with a wire loop -incubate at opt temp fro 24-48 hrs

Carbohydrate Fermentation Test - gas production?

-insert Durham tube which will capture some of the gas the organism produces

*Cestode*: Taenia saginata (beef tapeworm)

-intermediate host = cattle -mode of transmission = acquired by animals grazing on soil/vegetation contaminated by fecal wastes containing egg filled proglottids--these develop into larvae in animal intestines which migrate to muscle tissue and form cysticerci---man acquires by eating infected raw beef -symptoms/diagnosis = weight loss, abdominal cramps and gas -treatment/prevention = Niclosamide -may reach 36 feet *observe the scolex and proglottids*

*Cestode*: Echinococcus granulosis (Echinococcosis)

-intermediate host = dogs and cats -eggs of the worms released in stool & consumed by from direct contact w/animal or contaminated food; contact w/eggs on animal (infested dog) -hydatid cysts may form anywhere (brain, spleen, lungs) -surgery; Niclosamide *very short; look for scolex and proglottids*

*Trematode*: Fasciola hepatica (sheep liver fluke)

-intermediate host = fish; cercaria enter skin; causes severe liver damage -Bithional; avoid cercaria; Dehydroemetine (occurs in Asia, Australia) -adults release eggs in stool-contaminate water; eggs develop in snails; contaminate aquatic vegetation (sheep affected by consuming contaminated vegetation)

*Blood or Tissue Nematode*: Wucheria bancrofti (Elephantiasis)

-intermediate host = humans -mode of transmission = larvae of parasite are transmitted to new host by infected mosquitoes when feeds on bloody meal-travel to lymphatic vessels and into nodes where later develop into adults -swelling of extremities (edema); control mosquitoes; Diethylcarbamazine

*Cestode*: Taenia solium (pig tapeworm)

-intermediate host = pigs; man eats infected raw pork -first acquired by pigs consuming garbage or produce contaminated w/fecal waste w/egg filled proglottids-eggs hatch into larvae & migrate to tissue (now cysts) -muscle cysts; pain; cramps, and gas; Niclosamide *observe the scolex and proglottids*

*Trematode*: Schistosoma japonicum & mansoni (Adult Flukes) (Schistosomiasis & Bilharzia)

-intermediate host = snails -eggs hatch into miracidia & infect snails-reemerge as infective cercaria-man contacts cercariae by infected waters *a bird schistosome can cause "swimmers itch"* -intestines & liver; severe jaundice, abdominal pain, & anemia-Bithional; praziquantel

*Blood/Tissue Nematode*: Trichinella spiralis (Trichinosis)

-intermediate host-animals: pigs, bears, wolves -eating the encysted larvae in raw meat-cysts change to larvae in intestines, mature into adults and reproduce-eggs develop into larvae-enter lymph/blood/tissues -muscle pain, eye aches & fever; Mebendazole *note spiral worms in tissue section*

*Intestinal Nematode*: Trichuris trichiuria (whipworm or trichuriasis)

-intermediate host-humans; dogs; mode of transmission = larvae emerge from eggs, migrate to large intestine, & mature into adults--contact w/eggs on animals -diarrhea; abdominal pain; eggs or adult worms attached to mucosa -Mebendazole; good hygiene & sanitation; children at great risk (hand to mouth)

*Intestinal Nematode*: Enterobius vermicularis (enterobiasis or pinworms)

-intermediate host= humans (affects children) -poor hygiene; hand contact w/eggs-once consumed eggs hatch into adults in intestines; females migrate to anal region & deposit new eggs -anal itching; scotch tape test; Pyrvinium; Mebendazole; entire family needs tx *look for transparent oval sticky eggs*

*Intestinal Nematode*: Ascaris lumbricoides (Ascariasis or human roundworm disease)

-intermediate host= humans, dogs -consuming eggs that hatch into larva; later transform into adults; contact w/eggs on animals-complex life cycle - intestines to lungs back to intestines -abdominal pain-may become very active (crawl out); form obstruction -Mebendazole; Peperazine; citrate

Cryptosporidium parvum (Apicomplexa) DISEASE = Cryptosporidiosis

-intestinal infections especially in immune-compromised; diarrhea -contaminated water; community swimming pools -Nitazoxanide -opportunistic infections *trophozoite*

Why is it difficult to relate the effectiveness of a disinfectant tested in the laboratory to its effectiveness in actual use?

-it's difficult to simulate actual working conditions (too many variables) -need to control testing procedures to provide accurate comparative information

Protozoan Treatment & Prevention

-malaria prevented by quinine derivatives -amoebic dysentery, giardiasis, and trichomoniasis treated w/applications of metronidazole

In 21st Century America, why are we still concerned about tapeworm and roundworms infestations?

-many parasites utilize animal hosts -consequently, consumption of under-cooked, infected meat could result in infection -some parasites may also be found in fecal contaminated soil and water

Why do you suppose that meat samples can support a larger bacterial population than milk samples?

-meat provides all the available nutrients to bacteria -hamburger possesses more microbes because of the grinding process and the addition of air

Chiggers (vector = Trombicula mites)

-mechanical vector -their larva produce intense itching/inflammatory lesions

Head Lice (Vector = Pediculus Lice)

-mechanical vector spreading diseases such as impetigo and trachoma (treat w/insecticides) -itchy pest

Why are some arthropods mechanical vectors and others biological vectors?

-mechanical vectors have chewing mouth parts only and cannot transmit agents into the blood stream of their victims -biological vectors have piercing or sucking mouth parts which allow the agents access to the blood stream and/or access to the agents already in the blood stream (the agents can then use one victim for part of their life cycle and another victim for the remainder of their cycle)

The many variables accounted for in the Kirby-Bouer test [to prevent inaccurate results]

-medium used -diffusibility of antimicrobial agents through the medium -amount of antimicrobial agents used -number of microorganisms -time of incubation -measurement of zones of inhibition -interpretation of results

Nemathelminthes (roundworms or Nematodes)

-most numerous of multicellular parasites -characterized by having nervous and complete digestive system w/a true head, mouth parts, and anus -separate sexes and tough outer cuticle which resists intestinal enzyme degradation -ascarid roundworms produce large quantities of eggs that are released in stool of infected host

Are any parasitic diseases communicable from person to person?

-most parasitic diseases are not communicable -proper hand washing to avoid hand transmission is strongly advisable

Platyhelminthes (flatworms)

-most primitive of helminths -have a rudimentary digestive system w/only one opening and absorb food directly thru body wall -flattened dorsal-ventrally and shaped like a leaf (fluke) or tape measure (tapeworms) -have head region equipped with hooks and/or suckers

Transmission Electron Microscope

-most view in black & white -internal details are clearly visible -used to view viruses (b/c their very small)

Nitrate Reductase Test - positive confirmation test?

-no color change - organism reduced nitrate completely to ammonia and nitrogen gas

DNase agar - contents?

-nutrients for bacteria -DNA -methyl green -pH indicator that is a cation and will bind to the neg charged DNA

What are the laboratory procedures used for diagnosis and identification of protozoans?

-observation of intestinal parasites in concentrated stool samples -observation of blood parasites in blood samples -special serological lab tests

Helminth Diagnosis

-observation of larvae, adult worms, and/or eggs is indicative of infestation -stool, blood, or tissue samples may be tested -elevated eosinophil count is highly suggestive of active infection

Higher Parasites

-ones that affect humans commonly known as helminthes -possess organ systems -they have special attachment mechanisms such as hooks and suckers that allow them to adhere to host tissue; locomotion is minimal and depend on transfer from host to host by different means -Niclosamide used for tapeworm infections and Mebendazole for roundworm infections

Candida albicans (Yeast) disease = Candidiasis (mild skin infection) and thrush (oral) *note tiny oval yeast in center of field*

-opportunistic dimorphic yeast; Ascomycota -any entry route (found in GI and genitourinary tracts) -bartenders, dishwashers; immune compromised; overuse of antibiotics=yeast growth -oral/cutaneous infections=nystatin & antifungal creams; systemic infections=Amphotericin/azoles

Carbohydrate Fermentation Test - Acid Production?

-phenol red (pH indicator) -neutral - red -acidic - yellow

Antibiotic Sensitivity Testing

-plates swabbed with microbe prior to adding antibiotic disks -measure zone diameters (in mm) -compare clearing to chart to identify as: -resistant (R) -moderately sensitive (MS) -sensitive (S)

Could any protozoans be found normally on or in the human body? Which ones?

-possibly yes -E. gingivalis and Trichomonas hominus or tenax could be found in the oral cavity -their role in infection is not determined

Gram Stain Procedure

-prepare slide -drop of water -aseptic technique to add bacteria -heat fix -cool -add primary stain (crystal violet) and sit for 1 min -rinse with water -add mordant (Grams iodine) and sit for 1 min -decolorize with alcohol and sit for 30 sec -rinse with water -add counterstain (Safranin) and sit for 1 min -rinse with water -blot with absorbing paper

Of what importance is a protozoan cyst?

-protozoan cysts are very resistant to freezing, drying, polluted water, etc. -cysts provide a means for their transmission

How do the protozoans differ from bacteria?

-protozoans are eukaryotic, have a true nucleus, organelles, etc. -protozoans are much larger and more complex

Protozoan Cyst Formation

-resistant, dormant structures -covered by thick cell wall-like material called chitin which resists adverse conditions (freezing and polluted water) -form w/in intestinal tract of host and released in stool -consumption of food/water contaminated w/these will allow parasites to gain access to intestinal tract -they can then transform into free-living forms known as trophozoites

Giardia intestinalis or "lamblia" (Flagellate) DISEASE = Giardiasis

-severe prolonged intestinal dysentery -contaminated food/water- treat w/Metronidazole -cause intestinal infection (Beaver Fever); backpackers, hikers; maintained in wild animals *look for pear-shaped parasites with "eyes"-nuclei; can see trophozoites & very resistant cysts*

Why are some molds more capable of causing systemic infections than others?

-some are able to grow at 37 degrees C -some have thick cell walls that protect them from degradation and help them resist phagocytosis -some also have damaging extracellular enzymes -just the presence of fungi will cause a significant inflammation resulting in further tissue destruction

Why do organisms vary in their sensitivity to antibiotics?

-some organisms may possess plasmids which contain resistant genes -some organisms possess cell wall chemistry that is unaffected by the drug

Gelatin Liquefaction test - procedure?

-stab gelatin with organism using a needle -incubate at optimum temp for 24-48 hours -place tube in fridge for at least 30 minutes

Phenylalanine Deamination "Phenylaline Agar" - procedure?

-streak agar slant -incubate -add 5 drops of 10% Ferric Chloride on slant -wait 5 minutes

Compare the advantages and disadvantages of heat sensitive tape, chemical test strips, and endospore vials.

-tape = quick, easy to use, immediate information but limited sensitivity -test strips = quick, easy to use immediate, good sensitivity -endospore vials = easy to use, delayed results, certified test of sterility

Why are colonies found in the clear zones around an antibiotic disk important in the determination of sensitivity?

-the colonies represent resistant bacteria that formed into colonies -the antibiotic should not be used and the test should be recorded as resistant

Of what importance is antibiotic sensitivity testing to a physician?

-the test provides specific information as to the type of antibiotic that can be used in therapy -the physician is able to select the most appropriate drug that will be the most effective against the organism

Describe the difference between total count and viable count.

-total count = all microbes present, living or dead -viable count = only live microbes capable of producing colonies

A 1 milliliter sample of milk was diluted six times using 9 ml saline tubes. Duplicate samples were plated from the last three tubes. The plates were incubated and examined. The plate counts were: Tube 4 Tube 5 Tube 6 Plate A 812 116 26 Plate B 656 87 13 Determine the total number of bacteria per ml of original sample. *note: Tube 5 is used because the plate counts are between 30 and 300*

-total dilution = 1/10 X 1/10 X 1/10 X 1/10 X 1/10 X 1/10 -dilution factor = 100,000 PC (ave) = (116 + 87)/2 = 102 -total #/ml = 102 X 100,000 = 10,200,000 or 10.2 X 10^6

A 1 gram sample of hamburger was tested for bacterial count. The sample was first emulsified and diluted with a 99 ml bottle and further diluted using 5, 9 ml saline tubes. Duplicate samples were plated from the last three saline tubes. The plates were incubated and examined. The following plate counts were: Tube 3 Tube 4 Tube 5 Plate A: 792 220 17 Plate B: 985 175 28 Determine the total number of bacteria per gram of original sample. *note: Tube 4 is used because the plate counts are between 30 and 300*

-total dilution = 1/100 X 1/10 X 1/10 X 1/10 X 1/10 X 1/10 -dilution factor = 1,000,000 PC (ave) = (220 + 175)/2 = 198 -total #/gm = 198 X 1,000,000 = 198,000,000 or 19.8 X 10^7

Life Cycle of a Nematode

-transmitted to others via poor hygiene, flies, or feces (food/water) -after consumed, travel thru stomach to small intestine (hatch into tiny larvae) -penetrate mucous lining--enter the circulation--pass thru heart--into lungs -larval forms change in structure--travel up bronchi & trachea-coughed up & swallowed -finally end in intestinal tract again (where develop into adults)

Motility Test - motile positive?

-tube will appear red and cloudy -organism will spread over top of the medium

Cryptococcus Filobasidiella or Neoformans (Yeast) disease = Cryptococcosis

-type of mycoses = systemic -phylum = Basidiomycota -entry route = nasal -treatment = Amphotericin B and fluconazole -true yeast w/capsule -often disseminates to brain; transmitted from bird droppings (especially pigeons)

Coccidioides Immitis (Dimorphic) disease = Coccidioidomycosis (Valley fever)

-type of mycoses = systemic -phylum = Deuteromycota -entry route = nasal -treatment = Amphotericin B -in nature the fungus forms small arthospores which become airborne *arthrospores shown*

Histoplasma capsulatum (Dimorphic) disease = Histoplasmosis (Mississippi River Valley fever)

-type of mycoses = systemic -phylum = Ascomycota -entry route = nasal -infectious spores released in dust and inhaled by victim -one of most common respiratory fungal agents -treatment = Amphotericin B

Trematodes (flukes)

-un-segmented, covered by tough resistant cuticle, shaped like a leaf, and possess suckers -usually absorb food directly thru body wall -most flukes have separate sexes

Protozoa

-unicellular, eukaryotic organisms -only a few are of medical importance -generally unable to live independently of host and develop protective structures (cysts) to ensure survival -transmitted to host by vector (mosquitoes, flies, and ticks) -typically transmitted by contact w/human fecal matter -ectoparasites = those infecting skin/mucous membranes -endoparasites = those infecting internal organs or blood

Oxidase Test - indicator?

-uses an artifical electron acceptor -changes color to a dark blue/purple when its taken the last electron from cytochrome oxidase

Electron Microscope

-uses electron beams instead of light source: very short wavelengths so resolution and magnification are very good -views only DEAD specimens

Flourescence Microscope

-uses ultraviolet light source and "flourochromes" (flourescent stains) -helps quickly identify microorganisms: flourochomes naturally attach to specific microorganisms making them different colors

Claviceps purpurea (wheat rust)

-wheat fungus - forms black nodules called ergots -causes ergot poisoning (mycotoxicosis) -contracted by eating contaminated wheat and grains -poison causes severe circulation problems -may also be hallucinogenic

Opportunistic Mycoses

-when body's defenses against microbial invasion are weakened by malnutrition, cancer, radiation, or immunosuppressive drugs, normal flora initiate severe infections -examples: Candida albicans & Pneumocytis carinii

CITRATE TEST

..This test is among a suite of tests * Indole * (MR) Methyl-Red * (VP) Vogues-Proskauer * (Citrate) that are used to differentiate among the Gram-Negative bacilli in the family Enterobacteriaceae.

Normal % of basophils?

0.5-1%

Color indicators for VP testing

1 Rose Pink indicates the production of neutral end-products 2 No color change in the medium indicates

How many lobes do basophils have?

1 or 2

Normal % of eosinphils?

1-3%

The susceptibility of an organism to a drug is assessed by the size of the zone of inhibition, which is affected by:

1. Ability and rate of diffusion of the antibiotic into the medium and its interaction with the test organism 2. The number of organisms inoculated 3. The growth rate of the organism

Advantages of viable plate counts

1. Detects viable cells from dead cells 2. Isolate discrete colonies → pure colonies (helps facilitate identification)

2 forms of coagulase:

1. Free 2. Bound

Disadvantages of viable plate counts

1. Overnight incubation is time consuming 2. A lot of glassware is used 3. Greater manipulation→ increased counting errors

2 forms of the Coagulase Test:

1. Slide test 2. Tube test

Making a Capsule Stain

1. Tiny drop of INDIA INK (-) near on end of slide 2. Transfer BACILLUS MEGATERIUM into drop (do not smear with loop) 3. Use another slide to spread stain 4. Let air dry, heat fix, let cool 5. With slide on staining rack, flood smear with CRYSTAL VIOLET (+) and let sit for 1 minute 6. Tilt one end and rinse with water bottle 7. Blot dry 8. View under microscope OIL IMMERSION

Making a Negative Stain

1. Tiny drop of NIGROSINE near one end of slide 2. Transfer BACILLUS MEGATERIUM into drop (do not smear with loop) 3. Use another slide to spread stain 4. Let air dry 5. View under microscope

Making an Endospore Stain

1. Tiny loop full of water 2. Transfer both BACILLUS MEGATERIUM into water and smear with loop 3. Air dry, heat fix, let cool 4. Place slide on hot plate (including weigh boat and paper towel square on smear) 5. For 3-5 minutes, continuously keep the paper towel square saturated with MALACHITE GREEN while the slide gently heats 6. Remove from heat and place on staining rack to cool 7. Tilt one end and rinse with water bottle (decolorizer) 8. With slide on staining rack, flood smear with SAFRANIN for 30 seconds (rinse with water bottle) 9. Gently blot dry and view under microscope OIL IMMERSION

Making Acid-Fast Stain

1. Tiny loop full of water 2. Transfer both STAPH EPIDERMIS and MYCOBACTERIUM CHROMOGENICUM into water and smear with loop 3. Air dry, heat fix, let cool 4. Place slide on hot plate (including weigh boat and paper towel square on smear) 5. For 5 minutes, continuously keep the paper towel square saturated a bright pink color with CARBOFUCHSIN (+) while the slide gently heats 6. Remove from heat and place on staining rack to cool 7. Tilt one end and rinse with water bottle 8. With slide on staining rack, flood smear with ACID ALCOHOL decolorizer for 3-5 minutes (rinse with water bottle) 9. Flood smear with METHYLENE BLUE (+) for 30 seconds (rinse with water bottle) 10. Gently blot dry and view under microscope OIL IMMERSION

Making a Simple Stain (+)

1. Tiny loop full of water on slide 2. Transfer BACILLUS MEGATERIUM into water and smear with loop 3. Air dry, HEAT FIX, let cool 4. Flood smear with METHYLENE BLUE and let sit for 1 minute 5. Tilt one end and rinse with water bottle 6. Blot dry and view under microscope

Making a Gram Stain

1. Tiny loop full of water on slide 2. Transfer both STAPH EPIDERMIS and E. COLI into water and smear with loop 3. Air dry, heat fix, let cool. 4. With slide on staining rack, flood smear with CRYSTAL VIOLET (+) and let sit for 20 seconds 5. Tilt one end and rinse with water bottle 6. Flood smear with mordant Gram's IODINE and let sit for 1 minute (rinse with water bottle) 8. Flood smear with 95% ALCOHOL decolorizer and let sit for 10 seconds (rinse with water bottle) 10. Flood smear with SAFRANIN (+) and let sit for 20 seconds (rinse with water bottle) 11. Blot dry 12. View under microscope OIL IMMERSION

What are the 2 problems that occur with increasing magnification?

1. need for greater resolving power 2. decreasing image brightness

Capsule Stain

1.) negative stain with congo red 2.) air dry- don't heat fix 3.) Hydrochloric acid 4.) Safranin counter stain

How would a total dilution of 1:10,000,000 be prepared?

1/10 X 1/10 X 1/10 X 1/10 X 1/10 X 1/10 X 1/10

How would a total dilution of 1:2 X 10^4 be prepared?

1/20 X 1/10 X 1/10 X 1/10

What are the two reagents for VP test?

10 drops of a-naphthol and 10 drops of KOH

How soon can a nitrate test be read after the addition of the reagents?

10 minutes

If IgM is present...what does this mean?

1st response - recent infection

1. Salmonella 2. Shigella

2 medically significant Enterobacteriae

If IgG is present...what does this mean?

2nd response - past infection

1. Mannitol (carb) 2. NaCl (Sodium chloride/salt) 3. Phenol red pH indicator

3 ingredients in Mannitol/Salt agar

What solution is used for the Catalase test?

3% H2O2 solution (hydrogen peroxide)

Normal % of monocytes?

3-7%

at what temperature are cultures incubated in he detection of fecal coliforms

35C

1. Divide plate into thirds 2. Obtain swab of bacteria 3. Streak bacteria in zig zag in section one. 3. Drag from section one and zig zag in section two at 90 degree angle 4. Drag from section two and zig zag in section three at 90 degree angle from section 2.

4 Steps of Streak plate

Protozoan Movement

4 common types of movement = 1. pseudopodia- false feet allows members of Sarcodina (Endamoeba) class to extend protoplasm 2. flagella- long whip-like appendages that all members of class Mastigophora possess (Trichomonas) 3. cilia- short stiff bristle-like structures; members of the class Ciliata (Balantidium) 4. undulating membranes- Apicomplexa (Sporozoa); thin and undulate back and forth along one side; several members of Mastigophora (Trypanosoma)

Mnemonic: C CUT 1. Carbohydrate fermentation tests 2. Cytochrome Oxidase 3. Urease 4. TSI tests

4 tests frequently used to differentiate Gram negative bacteria.

What range should the rheostat be between so that the background of the image is white?

4-6

What total magnification is the high dry objective? What is the color of the ring?

400X/Blue

What total magnification is the scanner objective? What is the color of the ring?

40X/red

1. Prepare smear/heat fix 2. Cover with crystal violet (1 minute), rinse (distilled water) 3. Iodine mordant for 1 minute, rinse 4. Gram decolorizer (aprox 5-10 seconds), rinse 5. Gram safranin (30 sec), rinse and dry.

5 Steps of a Gram stain

BA - enriched with?

5% sheep's blood

neutrophil

55-90% phagocytes & killers of bacteria suggest cuts infections

AMC BOB 1. Amylase 2. Mannitol Salt 3. Catalase 4. Bacitracin 5. Optochin 6. Blood hemolysis

6 Tests commonly run on Gram positive bacteria

Phenol Red is yellow below what pH?

6.8

Phenol red is yellow below what pH?

6.8

Bromthymol blue is green at what pH?

6.9

How soon can the VP test be read after adding reagent?

60 minutes

What is the diameter of the lymphocytes?

7-18 micrometers

How big is a red blood cell? what can be seen by the naked eye?

7.5 microns/100 microns

MSA agar selective agent

7.5% salt-Sodium Chloride

Bromthymol blue is blue at what pH?

7.6

oil refractive index _______ glass refractive index (<,>,=)

=

The organism on this plate is resistant to Erythromycin.

??

Facultative anaerobe, negative ***Escherichia coli and Enterobacter aerogenes fit the description

A coliform is defined as a ______ that ferments lactose to produce gas, and it is gram _____. Which 2 baterica fits this description?

Station 11 Parasitic worms Description of disease

A disease of carnivorous mammals that infect the muscle

What are the color changes for Methyl Red?

Acid=red Neutral=orange Alkaline=yellow

What is the color changes for Citrate test?

Acid=yellow Neutral=green Alkaline=Blue

What are the color changes for phenol red during Carbohydrate Fermentation?

Acid=yellow Neutral=red-orange alkaline=cherry red

What turns the MacConkey agar red?

Acids from lactose fermentation

Station 12 Starch test Reagents added after incubation

Add indole to plate, stain starch after growth

Catalase is an enzyme found in:

Aerobes, facultative aerobes, microaerobes

Triple Sugar Iron (TSI)

Agar that contains dextrose, lactose, sucrose, phenol red for pH, and an iron indicator change in an agar slant.

Purpose of the Condenser Lens on Brightfield Microscope

Aims light through the specimen.

transmission of contagious diseases

Air droplets, direct contact with respiratory secretions or body fluids, through contact with contaminated inanimate objects

In experimentation why is it impossible to have an obligate anaerobe?

Although a THIO tube can provide 0% oxygen at the bottom of the tube, there is no way to innoculate the bacteria into the tube without exposing it to oxygen. Oxygen is present everywhere in the lab and oxygen is toxic to obigate anaerobe

Urease positive organisms will hydrolyze urea into:

Ammonia

What is the nitrogen source in Simmons citrate medium?

Ammonium dihydrogen phosphate

a. does not occur in the United States b. causes river blindness c. is transmitted by a biological vector d. *none of the above*

Amoebiasis: a. does not occur in the United States b. causes river blindness c. is transmitted by a biological vector d. none of the above

Station 12 Starch test Enzyme

Amylase

Bacteria secretes________ (enzyme) which breaks down______.

Amylase; starch

What is an oxygen requirement test and what is it made of?

An oxygen requirement test tests a microorganism on their need for oxygen. This determines if a microorganism needs oxygen to live, if oxygen is toxic to them or if it doesn't matter. It is made up of thioglycolate and methyene blue.

Bacillus anthracis

Anthrax

Antimicrobial produced by some molds and bacteria are generally called.

Antibiotics

Why would it be important to know the identity of a pathogen before prescribing an antibiotic?

Antibiotics are specific to certain bacterias; some organisms may be resistant to a certain antibiotic but others may be sensitive. Also, knowing the identity can allow the doctor to find the MIC and prescribe the right dosage to the patient so resistance doesn't occur.

Direct ELISA test for ....

Antigen

Station 10 Parasitic worms Location in humans

Any tissue in the body

Non-coliforms

Are Salmonella and Shigella coliforms or non-coliforms?

No

Are coliform bacteria usually pathogenic?

Yes

Are non-coliform bacteria usually pathogenic?

Station 10 Parasitic worms Stage passed to humans

Ate undercooked beef or pork.

What are two genera of endospore forming bacteria that are medically important?

Bacillus and Clostridium

Spore stain

Bacillus subtilis or Clostridium botulism

What is a spore-producing organism?

Bacillus subtilus

Streptococcus pneumoniae

Bacitracin resistant/ Optochin Sensitive bacteria

Streptococcus pyogenes

Bacitracin sensitive/ Optochin resistant bacteria

What is the difference between bacteria and eukaryotic microbes under a microscope?

Bacteria are significantly smaller and lack visible internal structures

E. coli

Bacteria that appears metallic green in color on EMB agar due to acid production

clear

Bacteria that do not ferment lactose on EMB agar appear ______

Alpha-hemolytic

Bacteria that do not make hemolysin, but do secrete chemicals that turn red blood cells a green color

Dark

Bacteria that ferment lactose on Eosin Methylene Blue (EMB) agar create colonies that appear ________

Staphylococcus aureus

Bacteria that is positive (turns agar yellow) in Mannitol/Salt agar test

what is a smear?

Bacteria that is spread on a slide and allowed to dry

Metallic green

Bacteria that produce a large amount of acid appear _________ on EMB agar.

Why is staining required to see bacterial cells

Bacterial cells are translucent. Staining is required so bacterial cells are visible under the microscope

What happens in the proccess of gelatin hydrolysis?

Bacterial species release proteases in the medium and "digest" the gelatin, so it is no linger to keep it's solid form and turns into a liquid.

This large protozoan is actually a good probiotic agent when consumed with yogurt and cheese and helps with digestion.

Balantidium coli

VP (Voges-Proskauer) test : Negative MR suggests that VP test may be performed in second tube . The reagents used for the VP test are

Barritt's reagent A&B . When these reagents are added to a broth in which acetyl methyl carbinol is present, they turn a PINK-BURGANDY color (a POSITIVE VP test).

tania sps.

Beef tape worm

blood agar with s. aureus

Beta hemolysis surrounding colonies. presumptive for s. aureus

Streptococci Group A & B

Beta hemolytic (complete destruction of red blood cells)

What does the amp resistance gene release?

Beta lactamase -hydrolyzes certain antibiotics

The Bacitracin test is used to differentiate:

Beta-hemolytic Strep. pyogenes and resistant vs. susceptible organisms

A dull green color; ***** Salmonella typhi produces a black or very dark brown color

Bismuth Sulfide Agar (BSA) test

Name some examples of epidemics

Black plague, smallpox, HIV/AIDS, ebola, influenza

Hydrogen Sulfide Production "Triple Sugar Iron Agar" positive result

Black precipitation

Fluroquinolones

Block DNA synthesis Ex: Ciprofloxacin

MR-VP TEST Theory : **MR-VP (Methyl Red- Voges-Proskauer)BROTH**media contains GLUCOSE and PEPTONE. All enterics oxidize glucose for energy; however the end products vary depending on bacterial enzymes.

Both the MR and VP tests are used to determine what end products result when the test organism DEGRADES GLUCOSE.

The Coagulase Slide test detects:

Bound coagulase

The Coagulase Tube test detects:

Bound or free coagulase

Differential for lactose/sucrose fermentation: **Lactose/ sucrose fermenting organisms = produce yellow/green colonies and turn surronding media yellow/green **Non - lactose/ sucrose fermenting organisms = produce opaque red/ pink/ white colonies and turn the surrounding media red

Brilliant Green Agar (BGA) test

pH indicators and respective medium and color produce?

Brom cresol Purple; Decaboxyl test; Purple @alkaline pH & yellow @acid pH

Name of pH indicator and its color at low and high/neutral pH for Snyfer test?

Brom cresol Purple; Neutral/high-green low/acid pH-- Yellow

pH indicators and respective medium and color produce?

Brom thymol Blue; Simmons Citrate Agar = Green @ neutral pH & Blue @ acid pH

pH indicator for Snyder test

Bromcresol green

What is the pH indicator of the Simmons Citrate Agar?

Bromothymol blue

What is the pH indicator for OF Glucose?

Bromthymol Blue

Station 9 Citrate test pH indicator

Bromthymol blue

What is a liquid medium called?

Broth

Station 6 Catalase test Positive result:

Bubbles.

Yeast produce by

Budding and fission

how are gram negative bacteria recolorized?

By using a counter stain (safranin, a red dye)

Describe how you would get the image in focus under oil immersion if you cannot see it the first time.

Bypass high dry, go back to low power, find object and focus. Go back to oil, add light, then find focus and it should be there.

C-I-A-S 1-1-5-1 P-S-D-C

C- Crystal Violet- 1 min -P- Primary stain I- Iodine- 1 min- S- Secondary Stain or mordant A- Acetone Alcohol- 5 sec- D- Decolorizer S-Safranin- 1 min- C-Counterstain

C-A- G-1-1 P-D-C ---------acid fast

C-carbolfuchsin Until Green Primary stain A- acid/Alcohol 1 min Decolorizer M- methylene Blue 1 min counter-stain

Glucose + Lactose + H2S + ????

Identify this KIA tube

Phenol red, Glucose, Lactose

In KIA test; Glucose and lactose fermentation is determined using a pH indicator ---------- which begins red and will turn yellow in the butt of the tube if ---------- is fermented and will cause the slant to turn yellow if ---------- is able to be fermented.

Source of infection

Index case

What is the reagent used for Oxidase?

Kovac's Oxidase

Candida

Little red clusters of berries

Chondrus crispus

Looks like sour kraut!

Paragonimus

Lung Fluke

MAC

MacConkey Agar

Vivax

Malaria

The substrate in MSA is:

Mannitol (carbohydrate)

MSA

Mannitol Salt Agar

MSA stands for:

Mannitol Salt Agar

PHENOL RED FERMENTATION TEST

Media : 0.5 % Glucose solution or 0.5% Mannitol broth with indicator phenol red . Procedure : broth is inoculated with test bacteria and incubated at 98F /37C For 24 hours and color noted.

Stab and Streak

Method used to perform a TSI agar test

MH

Mueller-Hinton agar plate

Station 17 Yeast and Molds What is the study of molds and yeast is called?

Mycology.

What are the results of MacConkey agar?

No Growth, Gram +, Lactose Unknown Growth, Gram -, Lactose - Growth, Gram + Lactose +

How do negative stains work?

Normally stains are positively charged and therefore attracted to the negatively charged bacteria. In a negative stain, the dye is negative and therefore repelled by the bacteria and the dye stains the surroundings/background

Station 1 Gelatin test Media

Nutrient gelatin (straw colored media/ purple cap)

Total Magnification

Ocular lens magnification (10) X objective lense magnification

Shigella

On SS Agar, which bacteria produces a colorless colony?

What does the VP test detect?

Organisms capable of fermenting glucose

What occurs in α-hemolysis?

Partial destruction of RBCs

MAC agar - NLF examples

Pathogens - Salmonella/Shigella "non coliform"

Streptococcus Pneumoniae

Pneumonia

In a Coagulase Slide test, clumping indicates:

Positive

In a SIM test, growth radiating out from the stab line indicates:

Positive motility

Antibody

Protein secreted by B lymphocytes

Penicillium

Purple branch with fingers

Trichomonas vaginalis

Purple dots WITH FLAGELLA!

What do both coagulase tests utilize?

Rabbit plasma (with anticoagulant)

Station 14 TSIA Forth tube

Red slant / Black butt - Glucose fermentation and sulfur reduction

+ Resistance

Resistant

What is an endospore?

Resting body formed by certain bacteria in response to harsh conditions that allows the bacteria to survive, not for reproduction

Station 16 Malaria Stage of development

Ring stage

Salmonella Typhi

Salmonella

Is MSA selective, differential, or enriched?

Selective due to sodium chloride Differential for S. aureus - fermentation

- Resistance

Sensitive

Citrate Utilization - agar?

Simmons Citrate Agar

Rhizopus

Spaghetti and Meatballs

Presumptive Test

Specific test for the detection of coliforms Lactose fermantation test (development of gas/acid is indicative of a coliform) MPN test gives idea of number of coliforms present

Staphylococcus Aureus

Staph infections, Toxic shock syndrome, Skin infections, MRSA

What organisms grow on MSA?

Staphylococci

Positive (clusters)

Staphylococcus tests __________ for Catalase, Nitrogen Reduction, and Arrangement tests

Staphylococcus epidermis

Staphylococcus that tests negative for Mannitol, Coagulase, DNase is what?

Staphylococcus aureus

Staphylococcus that tests positive for Mannitol, Coagulase, DNase is what?

Station 9 Citrate test Negative result:

Stays green/ usually no growth.

Hemolysin produced by Streptococci

Streptolysin O & Streptolysin S

Hemolysins produced by streptococci are called:

Streptolysins

Shigella

T-I-U for Shigella •TSI: Red/Yellow (lac-/glu+) •Indole ring in SIM Deep: Negative •Urease test: Negative •MacConkey Agar: COLORLESS bacause LACTOSE (-) Neg.

What are two important diseases caused by acid fast bacteria?

TB and Leprosy Mycobacterium tuberculosis and Mycobacterium leprae

Clostridium Tetani

Tetanus

Their only Carbon source

The Citrate test is used to detect bacteria that use citrate for this reason

Parfocal

The ability to change between lenses and still stay in focus

What is a titer?

The amount of antibodies in a serum

What does the MR test detect?

Organisms capable of performing mixed acid fermentation

CITRATE TEST THEORY

Organisms that UTILIZE CITRATE for energy produce ALKALINE compounds as by-products. Thus, a POSITIVE RESULT for citrate utilization is the formation of a BLUE color.

Brilliant Green Lactose Bile does what?

Oxgall and brilliant green are selective by inhibiting gram +. Lactose is differential ingredient.

E. coli

Oxidase negative bacteria example

Pseudomonas aeruginosa

Oxidase positive bacteria example

What is the effect of H2O2 on the cell?

Oxidizer

What is the purpose of mixing the reagents and the broth in the VP test?

Oxygenates the medium

When an epidemic becomes worldwide

Pandemic

These body lice are only annoying because they mess up our clothing.

Pediculus

What provides the protein in MR/VP broth?

Peptone

What is the pH indicator for Carbohydrate Fermentation?

Phenol Red

**Selects staphylococcus species since they can survive high salt concentrations ***Organisms capable of using mannitol as a food source will produce acidic byproducts of fermentation that will lower the pH of the media. The acidity of the media will cause the pH indicator, phenol red, to turn yellow *****Staphylococcus aureus is capable of fermenting mannitol (yellow) while Staphylococcus epidermidis is not (will remain red)

Phenol Red Mannitol Salt Agar (MSA)

Station 5 Urease test pH indicator

Phenol red

What is the pH indicator in TSIA?

Phenol red

PHENOL RED FERMENTATION TEST theory

Phenol red broth is a test medium to which a carbohydrate like glucose or mannitol is added . Phenol red is red under nuetral pH but tends to turn yellow under acidic condition. Acid production from fermentation of the sugar lowers the pH and medium turns yellow indicating carbohydrate utilization by the bacteria . (A) Gas production also from fermentation can be detected as a bubble in an Inverted Durham's tube inside the broth. (AG)

Major Groups of Antiseptics and Disinfectants

Phenolic compounds Alcohols Halogens Heavy metals Surface-active agents Acids/Alkali Ethylene Oxide B-propiolactone Basic dyes

Ascaris

Pig roundworm

Gram Stain

Pink = gram - purple = gram +

What color is Gram -?

Pink/fuchsia/colorless

Enterobius vermicularis

Pinworm (Dead Bean)

Station 16 Malaria Causative organism genus

Plamodium

This parasite can be transmitted by the Anopheles mosquito.

Plasmodium falciparum, malariae, & vivax

What was the control plate in this UV experiment?

Plate with lid

Station 10 Parasitic worms Phylum

Platyhelminthes

A blue result in a citrate test indicates:

Positive

A darkened medium in a Bile Esculin Test indicates:

Positive

A liquid result for a gelatin test indicates:

Positive

A red result for VP indicates:

Positive

A red result for the MR test indicates:

Positive

Bubbles present in a catalase test indicate:

Positive

In a Coagulase Tube test, a solid medium indicates:

Positive

Dark blue color on an oxidase test is indicative of:

Positive (Cytochrome c oxidase present)

In a SIM test, blackening of the medium indicates:

Positive (sulfur reduction)

A pink result for a urea test indicates:

Positive (urease)

Citrate Test

Positive organism= E.coli Positive color results= Blue Indicator= bromthymol blue carbon source= Sodium citrate nitrogen source= ammonium dihydrogen phosphate

Station 8 Methyl Red test Pick the positive tube

Positive result: Red in color, means : glucose was broken down into various acid products

Interpretation and Potential bacteria for Urea

Positive-Proteus-instense pink; E. coli-yellow-Negative

What resists color changes in the MR/VP tube?

Potassium phosphate

What does the oxidase test detect:

Presence of cytochrome c oxidase

What does a gelatin hydrolysis test detect?

Presence of gelatinases

Gram Stain - 3 stains

Primary - crystal violet Mordant - Iodine Counterstain - Safranin

Acid

Product of carbohydrate fermentation

How does blackening of the medium occur?

Production of H2S gas combines with iron, to make ferric sulfide (FeS) which is a black precipitate

Positive ***Proteus contains cystein desulfurase enzyme that produces H2S in the presence of iron salts (also noted in KIA and SIM test)

Proteus is H2S (Hydrogen sulfide test) positive or negative?

Postive on the Urea test

Proteus is Urease positve or negative?

Causes of malaria

Protozoan plasmodium species

These organism are typically oxidase positive:

Pseudomonadaceae

Pseudomonas

Pseudomonas gives NEGATIVE Voges Proskauer Indole and Methyl red tests, but a POSITIVE catalase = Liberation of Bubbles and oxidase test = Blue purple color

Most resistant bacterium:

Pseudomonas; Serratia may also be very resistant

After culturing, my unknown bacteria on a HEA agar in an incubated 37 degrees celsius and no growth

Pseudomopheda Floresess

Saccharomyces

Purple Honeycomb

What color is present when starch has been not been hydrolyzed? is is + or -?

Purple and it is starch hydrolysis (-)

What color is Gram +?

Purplish/red

Breaks down urea to ammonia and CO2

Purpose of urease enzyme

What is the mode of UV for microbial killing?

Pyrimidine Dimer

A slide from Chronic Lymphocytic Lukemia patient would contain what two cells?

RBC (Erythrocytes) and Lymphocytes

Iodine

Reagent for amylase test

Hydrogen peroxide

Reagent for catalase test

Kovak's reagent

Reagent for cytochrome oxidase test

Phenol Red is what color between 6.9 and 7.3?

Red

Station 4 Nitrate test Important

Red before zinc - positive Colorless after zinc - positive Red after zinc - negative

Station 4 Nitrate test Positive result :

Red color after A and B- have nitrate in the tube If colorless add zinc after all 3 steps, if still colorless its positive.

Station 4 Nitrate test Negative result:

Red color after adding zinc

Station 14 TSIA Fifth tube

Red slant / Black butt - Glucose fermentation and sulfur reduction

Station 14 TSIA First Tube

Red slant/yellow butt, and agar lifted - Glucose fermentation with acid production. Proteins catabolized aerobically (in the slant) with alkaline products (reversion), also produced gas

Station 2 Phenol Red media Positive result:

Red/Pink - no carbohydrate fermentation was used but proteins were used to grow

Station 3 Indole test Positive result:

Red/pink ring at the top, means that the bacteria produced tryptophanase and breakdown the substrate tryptophan, one being indole.

4 Lenses Magnification

Red: scanning (4x) Yellow: low power (10x) Blue: high power (40x) White: oil immersion (100x)

Purpose of the Diaphragm Adjustment Lever on Brightfield Microscope

Regulates the amount of light passing through the specimen.

The continual source of infection

Reservoir

Hydrogen Sulfide Test

SIM agar -- semi soft to detect motility Tube C is positive for H₂S production

On this medium Salmonella usually produces a black colony, Shigella a colorless colony & all lactose positive colonies appear red

SS Agar test

Hydrogen Sulfide Production "Triple Sugar Iron Agar) Bacteria

Salmonella & Proteus

SS Agar

Salmonella produces a black colony on which medium?

Bismuth Sulfide Agar (BSA)

Salmonella typhi produces a black or very dark brown color in which test medium?

Epidemiology

Science that deals with when and where diseases occur and how they are transmitted

Epidemiologist

Scientist that complies the data and tries to locate the source of the infection

Is Bile Esculin Agar selective, differential, or enriched?

Selective due to bile Differential for enterococci and Strep. bovis

Is MacConkey agar selective, differential, or enriched?

Selective due to bile salts and crystal violet Differential for lactose fermenters

Is Eosin Methylene Blue agar selective, differential, or enriched?

Selective due to dyes Differential for lactose fermenters

MacConkey Agar

Selective for detection and isolation of gram (-) organisms and differential for gram negative enteric bacilli Inhibits growth of staphylococcus, enterococcus, and streptococcus

Great inhibition zone

Sensitive

When doing Antibiotic Susceptibility test what inoculation tool and technique is used?

Serile swab/Bacterial Lawn

DNase agar - positive test example?

Serratia marcescens

Fasciola hepatica

Sheep liver fluke

Shigella Dysenteriae

Shigellosis

Beta hemolytic organisms

Show complete hemolysis, giving a clear zone with a clean edge around the colony **pathogenic

Alpha hemolytic organisms

Show incomplete hemolysis, producing methemaglobin and a green, cloudy zone around the colony

Gamma hemolytic organisms

Show no hemolysis and no change in the blood agar around the colony occurs

CITRATE TEST media

Simmon's Citrate agar slants contain sodium citrate (the only carbon source) and ammonium ion (the sole nitrogen source). A pH indicator, Bromothymol Blue is also included. Bromothymol Blue is GREEN at pH < 7.0 and BLUE at pH > 7.6.

Station 9 Citrate test Media

Simmons Citrate (green slant/navy Cap)

Medium used for Snyder test

Snyder test medium

What is the source of sulfur in a SIM medium?

Sodium thiosulfate

Which media gives the best isolation and is easiest to see if contamination has occured?

Solid media

Station 9 Citrate test Principle

Some bacterias can use citrate as its only source of carbon, some cannot.

Escherichia coli

Some microorganisms like _________, produce a metallic green sheen on Eosin Methylene Blue (EMB) test.

Hydrogen Sulfide Production "Triple Sugar Iron Agar" inocculation procedure

Stab/Streak

•G+ cocci •on BAP media--colony morphology: MEDIUM sized WHITE colonies; NO hemolysis •Microscopic Morphology: staph = clusters •part of normal flora on human skin •LOVES prosthetic devices •opportunistic pathogen •catalase + •coagulase -

Staphyloccocus epidermidis Colonies are circular, convex with an entire margin.

boils are caused by

Staphyloccus aureus

**Same is MSA Mannitol test but lacks phenol red indicator. **Selects for Staphylococcus and allows for development of natural colony pigment formation unlike in MSA

Staphylococcus 110 Medium test

What gram positive bacteria would result in a positive catalase test and as well as having bright yellow colonies in a mannitol salt agar

Staphylococcus Aureus

DNase agar - negative test example?

Staphylococcus aureus

The Coagulase Test is used to differentiate?

Staphylococcus aureus

What organism ferments mannitol?

Staphylococcus aureus

Gelatin Deep- visual reaction results

Staphylococcus aureus - Liquefied Positive Serratia Marcescens - Positive Bacillus Cereus - Positive Micrococcus roseus - Solid Negative Micrococcus luteus - Negative Bacillus subtilis - Negative Escherichia Coli - Negative Enterobacter aerogenes - Negative Proteus vulgaris - Negative

•Staphyle = clusters ("grape-like clusters") •Large golden colonies •Creamy appearence on plate •Narrow beta-hemolysis zone around colonies

Staphylococcus aureus:

BA - gamma hemolysis example

Staphylococcus epidermis "normal flora"

CATALASE TEST

Staphylococcus, Micrococcus and most aerobic organisms produce catalase Streptococcus and most anaerobic bacteria are UNABLE to produce it.

Station 12 Starch test Substrate

Starch

Station 12 Starch test Media

Starch plate

1. Use swab to fill 3/4 of plate 2. Turn plate 1/4 turn, turn swab, repeat fill 3. Turn plate another 1/4 turn, turn swab, repeat fill

Steps of making a spread plate

What technique is used to encourage streptolysin activity on a blood agar plate?

Streak-stab Technique

CATALACE what organism is beta hemolytic pattern (clear)?

Strep

Streptococcus pyogenes

Strep throat, scarlet fever, rheumatic fever

What Gram-positive organisms are catalase negative?

Streptococcus

Gram negative, cocci

Streptococcus and Staphylococcus are both gram ______ and as shaped as _______.

BA - incomplete hemolysis example

Streptococcus pneumonia

The Optochin test is used to differentiate:

Streptococcus pneumoniae

What's Optochin sensitive?

Streptococcus pneumoniae

BA - complete hemolysis example

Streptococcus pyogenes -Strep Throat

scarlet fever is caused by

Streptococcus pyrogenes

Alpha and beta

Streptococcus shows what type of hemolysis?

Negative (chains)

Streptococcus tests __________ for Catalase, Nitrogen Reduction, and Arrangement tests

Streptococcus pneumoniae

Streptococcus that causes alpha hemolysis in blood agar is what?

Streptococcus pyogenes

Streptococcus that causes beta hemolysis in blood agar is what?

-gain entrance to body as result of trauma to skin -most are common soil inhabitants which enter skin through splinters, thorns, or sphagnum moss -example: Sporothrix schenckii

Subcutaneous Mycoses

Mannitol

Sugar in Mannitol/Salt agar

Lactose

Sugar source in EMB agar

What is the biochemical reaction for sugar fermentation?

Sugar---------------------> acid with/without gas

In a TSIA slant, blackening of the medium indicates:

Sulfur reduction

In a TSIA slant, a yellow slant/black butt indicates:

Sulfur reduction and glucose and lactose/sucrose fermentation

In a TSIA slant, a red slant/black butt indicates:

Sulfur reduction and glucose fermentation

How is ferrous sulfide produced in the hydrogen sulfide production test?

Sulfur-containing amino acids (already in the tube) are broken down into hydrogen sulfide and mixed with iron sulfate (already in the tube) to produce ferrous sulfide

When doing Antibiotic Susceptibility test what observation are made in Lab and what reported out to physicians?

Susceptibility & Resistance

In a Bacitracin test, a zone of clearing greater than 10 mm indicates:

Susceptible

In a Novobiocin test, a zone of clearing greater than 16 mm indicates:

Susceptible

In an Optochin test, a zone of clearing greater than 14 mm indicates:

Susceptible

Proteus

T-I-U for Proteus spp. •TSI: Red/Black (lac-/glu+/H2S+) •Indole: depends on species--p. vulgaris (+); p. mirabilis (-) •Urease: Strong Positive that results in magenta color •MC: COLORLESS because lac- •PUTRID, terrible, NASTY smell

•TSI: Red/Black (Slight yellow in the bottom of butt) (lac-/glu+/H2S+) •Indole: Negative •Urease: Negative •MC: COLORLESS because lac- •Hektoen Enteric Agar: Black precipitate, reaction between ferric ammonium citrate and H2S produced from SULFUR reaction

T-I-U for Salmonella:

In lab 5 (identifying an unknown), which mediums needed to be innolculated with a needle?

THIO tube, SIM tube and gelatin

Station 10 Parasitic worms Genus

Taenia

Station 10 Parasitic worms Species of pork tapeworm

Taenia Sollium

Station 10 Parasitic worms Species of beef tapeworm

Taenia saqinata

These worms possess a head end called a scolex and reproductive units called proglottids.

Taenia solium & saginata (platyhelminthes; cestodes)---tapeworms

•Indicates the ability of the bacteria to produce the enzyme urease, which converts urea → NH3 (ammonia) + CO2 •Color change due to pH change (alkaline) •Yellow (acidic) to Rose (alkaline) or Magenta (alkaline) •Strength of pink color is dependent upon how much ammonia is produced

Test #3-Urease Test:

Confirmed Test

Test that requires EMB media (selective for gram neg) Coliform colonies are metallic green

Bacitracin & Optochin (could also use hemolysis/blood agar)

Test used to differentiate between Streptococcus pyogenes and Strep. pneumoniae

Completed Test

Test where colony is inoculated into lactose broth A gram stain is also prepared -Tubes with lactose + and gas/acid further confirm presence of coliform bacteria -gram (-) bacilli

catalase test

Tests for catalase enzyme. adding H2O2 to culture results in Bubble formation = positive. negative = no bubble formation

Acid Fast Stain

Tests for mycolic acid. Carbolfushin is a lipid soluble stain, and thus penetrates. Counter stain with methylene blue.

Streptococcus pyogenes

The McKesson Strep A dipstick test is a rapid chromatographic immunoassay for the detection of Strep A antigen. What bacteria does this test identify?

Gram Negative Enterics

The Methyl Red/Voges-Proskauer Test differentiates between these types of bacteria

Streptococcus mitis

The bacteria on this plate are called?

Streptococcus salivarius Sucrose into glucose and fructose Tongue

The bacteria on this plate are called? What do they breakdown? Where are they found?

Streptococcus mutans Ferment fructose into lactic acid Teeth

The bacteria on this plate are called? What do they ferment? Where are they found?

Penicillin

The bacterial species on this plate have been incubated with antibiotic disks. Which antibiotic is the bacteria least sensitive (most resistant) to?

Ciprofloxacin

The bacterial species on this plate have been incubated with antibiotic disks. Which antibiotic is the bacteria most sensitive (least resistant) to?

Diameter of field

The distance across the entire visible field when looking through the ocular lenses.

Dinoflagellates

The little monster

Minimum Inhibitory Concentration (MIC)

The lowest concentration of an antimicrobial that will inhibit the visible growth of a microorganism (creates a zone of inhibition)

Purpose of Course Adjustment Knob on Brightfield Microscope

The main focusing tool.

a. are all members of the animal kingdom b. only cause respiratory diseases c. are all unicellular d. *are commonly classified on the basis of their sexual spore formation*

The mycotic agents: a. are all members of the animal kingdom b. only cause respiratory diseases c. are all unicellular d. are commonly classified on the basis of their sexual spore formation

How does a halo form in the starch hydrolysis test?

The presence of amylase will decompose starch into glucose molecules. Iodine only reacts with starch to make it a brown color. So, if amylase is present and breaks down all the starch, a yellow halo will appear

Transformation

The process by which competent bacterial cells take a DNA to form recombinant DNA

INDOLE RING TEST procedure

The test organism is inoculated into tryptone broth, a rich source of the amino acid tryptophan. Indole POSITIVE bacteria such as ESCHERICHIA COLI produce tryptophanase, an enzyme that cleaves tryptophan, producing indole and other products. Tubes are incubated at 98F/37C for 24 -48 hours .

why are agar plates used when observing microorganims?

They provide a large surface area for the ease in observing or retrieving many individual colonies

What is the point in observing untreated ("time zero") plates in an experiment where you're observing the effective powers of heat and UV light treatments?

They serve as a control to compare the other times to.

Why is thioglycolate used in an oxygen requirement test?

Thioglycolate eats up oxygen, so it creates a gradient in which oygen is about 21% at the top of the tube and 0% at the bottom.

Enterococcus It is not a coliform MacConkey Agar

This agar is negative for what? Why? What agar is it?

Proteus It is not a coliform MacConkey Agar

This agar is negative for what? Why?What agar is it?

E. coli CHROMagar

This agar is positive for what? What agar is it?

Enterococcus CHROMagar

This agar is positive for what? What agar is it?

Klebsiella CHROMagar

This agar is positive for what? What agar is it?

Proteus CHROMagar

This agar is positive for what? What agar is it?

E. coli It is a coliform MacConkey Agar

This agar is positive for what? Why? What agar is it?

Klebsiella It is a coliform MacConkey Agar

This agar is positive for what? Why? What agar is it?

Entamoeba histolytica

This amoeba may contaminate food or water and cause a form of dysentery.

Bacillus cereus

This microbe forms gram-positive rods. Endospores may be visible in older cultures as clear areas inside the cells. Older cultures may stain pink, due to the deterioration of cell walls.

Aspergillus Species

This mold may contaminate peanut butter, peanuts, rice, and grains.

AB-

This person has what type of blood?

B-

This person has what type of blood?

O+

This person has what type of blood?

Claviceps purpurea

This wheat rust is actually good to eat in cereal along with blue berries and sugar.

Candida albicans

This yeast can affect bartenders and dishwashers and cause an inflamed skin infection.

Canida abicons

Thrush and moniliasis (vaginal yeast infections)

why is the disk-diffusion method used?

To determine the effectiveness of some antibiotics on certain bacteria

What's the purpose of the HardyCHROM Biplate Experiment (Hurbi)?

To diagnose a urinary tract infection & isolate/differentiate bacteria with in the urine.

Function of immersion oil

To reduce scattering of light which improves resolution

What is the purpose of an inverted Durham tube?

To trap a portion of any gas produced

Pregnant women should avoid cleaning out the kitty litter box to avoid contact with this parasite.

Toxoplasma gondii

This bug actually transmits parasites by biting the victim and then depositing the parasites in its fecal matter into the bite wound.

Triatomid Bug

Station 11 Parasitic worms Genus

Trichinella

This worm causes a sexually transmitted disease.

Trichomonas vaginalis

This parasite causes a vaginitis that can be treated with metronidazole.

Trichomoniasis vaginalis

Station 11 Parasitic worms Species

Trichonoma Spiralis

These worms can be sautéed up in butter and served with meat balls along with a good Chianti wine.

Trichuris trichiura

TSIA stands for:

Triple Sugar Iron Agar

True/False: Bilharzia is a major world health problem because many lakes and streams are infested with the flukes that cause this disease.

True

True/False: Diagnosis of intestinal parasites can often be done following an O&P check.

True

True/False: Humans can be exposed to Histoplasmosis because it affects birds and chickens.

True

True/False: Metronidazole can be used as a treatment for many intestinal protozoan infections.

True

True/False: The Mastigophora exhibit a method of movement with flagella.

True

True/False: The disk diffusion method (Kirby Bauer) is one of the most common methods for the determination of antibiotic sensitivity of bacteria.

True

True/False: The most common treatment for tapeworm infections would be niclosamide.

True

True/False: Trypanosoma cruzi causes Chagas disease which occurs in Central and South America.

True

False

True/False: All fungi are classified on the basis of their asexual means of reproduction.

True

True/False: Special culture media is required to grow the fungi and yeast because they grow slower than bacteria.

This parasite occurs most commonly in Africa and is transmitted by a tsetse fly.

Trypansoma brucei gambiense & brucei rhodesiense

Station 3 Indole test Substrate

Tryptophan

What must be present for the production of indole?

Tryptophan

Indole Test

Tryptophan ----> Tryptophanase ----> Pyruvate

Station 3 Indole test Enzyme

Tryptophanase

Urea production: Negative result, Positive result, media used

Tube remains a peach color, tube turns hot pink (ammonia production), urea broth

Mycobacterium Tuberculosis

Tuberculosis

Beta-hemolytic

Type of bacteria that make hemolysin

Selective and Differential

Type of media Eosin Methylene Blue (EMB) agar is

Differential

Type of media Hemolysis/blood agar test is

Phenol red

Type of pH indicator in Mannitol/Salt agar

Mannitol Salt agar, Yellow = +, Staphylococcus aureas = +

Type of test? Color that = positive result? Example positive

Amylase Test. Clear + positive.

Type of test? What indicates a positive result?

Cytochrome Oxidase Test. B is +.

Type of test? What is positive?

Simmons Citrate, B = +

Type of test? Which tube is +?

Urease test. Right tube (Pink) = +.

Type of test? Which tube is +?

Obligate aerobes, Facultative anaerobes

Types of bacteria that produce catalase

what happens to DNA when UV light is exposed?

UV light changes DNA that result in mutations in living organisms. these induced mutations can cause cell death.

Station 11 Parasitic worms Stage passed to humans

Undercooked meat containing juvenile nurse cell eaten

Urease Test

Urea ----> Urease ----> Ammonia

pH indicators and respective medium and color produce?

Urea Broth; Phenol Red; Intense pink

What nutrients are found in urea broth?

Urea and yeast extract (trace)

Station 5 Urease test What is the product that turns it a pretty pink?

Urea hydrolysis produces ammonia, which raises the pH in the medium and turns the pH indictor pink.

coarse adjustment knob

Used for focusing with 4x and 10x objective lenses

Methyl Red Test

Used to differentiate between E. coli and E. aerogenes pH indicator detects concentration of acid end products (+) red pH 4 -- E. coli (-) yellow pH 6 -- E. erogenous

Negative Staining (general)

Uses acidic (-) stain which colors the background and not the cells. -would use in Cerebrospinal fluid specimens to look for fungi)

Simple Staining (general)

Uses one basic (+) stain which colors cells

Differential Staining (general)

Uses two different colored basic stains which helps differentiate between types of cells.

What Reagent is used for MRVP? What is the pH indicator?

VP Reagent A , VP Reagent B Methyl Red

What's Optochin resistant?

Viradans: Streptococcus mutants

Hemolysis

Virulence factor of some bacteria that aids in differentiating types of Gram + bacteria on blood agar

Red tube = +. Acetoin & 2,3 butandiol

Voges-Proskaeur test. Which one is positive? For what is it positive?

What are we doing to the E.coli bacteria in the transformation experiment?

We are making the bacteria competent by making the negative bacteria accept DNA which is negative

UV-A-longest UV-B-medium length UV-C-shortest

What 3 groups does ultraviolet light divide into?

Neutrophil Granulocytes that are the first response to defend against bacterial and fungal infections

What WBC is this? What is its function?

Basophil Granulocyte that is important allergic** and parasitic resposnes

What WBC is this? What is their function?

Eosinophil Granulocytes that destroy parasitic worms, also involved in allergic responses

What WBC is this? What is their function?

Lymphocyte Initiate the immune response

What WBC is this? What is their function?

Monocyte Phagocytes that remove debris, phagocytose pathogens

What WBC is this? What is their function?

anti-B

What antibodies would agglutinate this antigen?

Intestinal bacteria that are gram (-), non-spore-forming bacilli that ferment lactose to acid and gas

What are coliform bacteria?

Part of normal flora Lactose fermenters Gram negative

What are coliforms?

Gram (+), non-spore-forming bacilli that do not ferment lactose to acid and gas

What are non-coliform bacteria?

Lactose non fermenter Not part of normal flora

What are some characteristics of enteric true pathogens?

Fermenting lactose or sucrose Enteric coliform

What are the bacteria doing on this plate? What organisms are growing on this plate?

They are not fermenting lactose or sucrose Non-coliform

What are the bacteria doing on this plate? What organisms are growing on this plate?

- gram negative - normal enteric flora - do not ferment lactose - opportunistic infections

What are the characteristics of NonColiform bacteria?

Proteus

What are the colonies surrounded by the brown halo on this plate called?

Blue = oxygen, white = NO oxygen.

What are the colors inside the anaerobic jar indicating?

Sexual contact, transfer of infected blood, infected needles, breast milk, during labor.

What are the modes of transmission of HIV?

Ingestion Inhalation Direct contact Animal bite Blood transfusion Sexual contact Sick people or animals Carriers of disease (animal or human) Contaminated food, water, objects Aerosols

What are the modes of transmission of infectious diseases?

E. coli

What are the red colonies shown on this plate called?

Klebsiella

What are the steel-blue colonies on this plate called?

Flu-like for 2-4 weeks, then asymptomatic for 10 yrs. Fevers & sweats from infections, weight loss.

What are the symptoms of HIV?

Enterococcus

What are the turquoise colonies on this plate called?

Staphylococcus epidermidis

What are the white colonies on this MSA plate?

Micrococcus luteus

What are the yellow colonies on this MSA plate?

Staphylococcus aureus

What are the yellow colonies on this MSA plate?

Do not rinse and do not heat fix

What are two things you should NOT do when doing Negative staining?

Formation of ammonia when bacteria ferments citrate

What causes that bromothymol blue to turn blue in a citrate test?

Alkaline pH of ammonia

What causes the pink color in urease agar?

Red, opaque colonies

What color do coliforms grow as on the MacConkey and CHROMagar?

Acidic- dark blue colonies with a metallic green sheen Less acidic- pink colonies

What color do fermented bacteria become on EMB agar?

Colorless

What color do non-fermented bacteria become on EMB agar?

Translucent colonies

What color do non-lactose fermenters grow as?

Dark colonies with a metallic green sheen OR pink

What color does E. coli and Klebsiella produce on EMB plate?

Clear

What color does Proteus and Enterococcus produce on EMB?

Pink

What color is gram negative?

Purple

What color is gram positive?

Toxin A and B antigen produced by Clostridium difficile

What did we test for using indirect ELISA?

Serum antibodies in blood against HIV.

What do both ELISA & Western blot detect?

Distort bacteria DNA, inhibit replication and transcription.

What do these pyrimidine dimers do to DNA?

Means to retain the dye when challenged by a decolorizer.

What does "fastness" mean in terms of acid fast staining.

AIDS-Acquired Immunodeficiency Syndrome

What does HIV cause?

Protein A present only in staph. Staph aureus gives clumping aka agglutination.

What does Ig G bind to?

A second antibody is added which binds to the serum antibody against HIV. A chromogen is added to bind to the antibodies added resulting in coloring, coloring shows a positive result.

What does a positive HIV result for ELISA show?

Fibrinogen & Ig G

What does the Catalase test use?

Lactose Fermentation only

What does the EMB plate assess?

CD4 T helper cells causing them to drop

What does the HIV virus attack?

Assess H2S production & Lactose fermentation of gram- enteric bacteria.

What does the Hektoen agar assess?

Bacterial cell wall synthesis

What does the Optochin disk inhibit?

Chemical pack and water

What does the anaerobic jar have inside?

New covalent bonds between adjacent pyrimidines (c-c, t-t, c-t), aka pyrimidine dimers.

What does the energy in UV light form?

Staphylococcus Aureus Causes yellow color due to fermentation of manitol. pH=6.8

What gram positive bacteria is growing on this Manitol media and why?

She will produce antibodies against Rh which will cross the placenta during the 3rd trimester. They will attack the fetus, causing a miscarriage (usually only with the second pregnancy)

What happens if a pregnant woman is Rh (-) while her baby is Rh (+)?

Impetigo, Cellulitis, abscess, boils, furuncle, food poisoning, toxic shock syndrome, septicemia.

What illnesses are caused by Staph Aureus?

Sepsis, scarlet fever, Rheumatic Fever, Necrotizing Fasciitis, Toxic Shock Syndrome, septicemia, nephritis

What illnesses are caused by Streptococcus species?

Methyl red

What indicator does the Voges-Proskauer test use?

Dyes

What ingredient in Eosin Methylene Blue (EMB) inhibits the growth of Gram Positive bacteria?

To differentiate organisms that may infect the urinary tract E. coli- red colonies Klebsiella- steel blue colonies Enterecoccus- turquoise colonies Proteus- brown halo

What is CHROMagar used to determine?

Whether or not bacteria ferment lactose to acid and gas

What is EMB and MacConkey Agar used to determine?

Grows with or without O2 Detoxify toxic O2 byproducts.

What is a facultative anaerobe?

Fomite may be defined as an inanimate item that could transfer microbial organisms. Ex: a fork

What is a fomite?

Requires small O2 amount Detoxifies O2 byproducts Does not grow in atmospheric O2

What is a microaerophile?

Can't live in oxygen Can't detoxify byproducts of O2.

What is an anaerobe?

The study of causes, occurrence, transmission, distribution & prevention of disease

What is epidemiology?

Masses of bacterial cells, dextran, and food debris

What is plaque?

Transmission from person to person

What is propagated transmission?

Differentiating between Staph Aureus and Staph Epidermis.

What is the Catalase test used for?

Detecting HIV, Enzyme Linked Immunoabsorbant Assay

What is the ELISA test used for?

Identifying enteric bacteria

What is the Enterotube test used for?

High O2 at top decreasing to little O2 on bottom.

What is the O2 gradient on the test tube used to grow these organisms?

Incidence: Number of newly infected people Prevalence: Number of infected people alive.

What is the difference between incidence rate and prevalence?

HIV proteins separated by size, transferred to a membrane, then serum antibodies, conjugate and chromogen are added, positive = spots

What is the difference in the process of ELISA vs. Western blot?

# of new cases at specific time/ population at risk multiplied by K

What is the formula for Incidence rate?

# of existing cases at specific time/total population multiplied by K

What is the formula for prevalence?

Remove O2 and maintain complete anaerobic atmosphere inside

What is the function of the anaerobic jar?

The largest clear zone where the microorganism was inhibited.

What is the minimum inhibitory concentration?

Index case

What is the name for the first detected case?

Balantidium coli

What is the name of this organism?

1:32

What is the patients titer?

Chemical pack reacts with Water pack to make anaerobic condition.

What is the process occurring in the anaerobic jar?

1:4

What is the result for this test?

Coccidioides immitis; spherules

What is this organism and feature?

Giardia intestinalis; cyst

What is this organism and phase?

Entamoeba histolytica

What is this organism?

Enterobius vermicularis

What is this organism?

Enterobius vermicularis; as an adult pinworm

What is this organism?

Necator americanus

What is this organism?

Trichinella spiralis

What is this organism?

Trichomonas vaginalis

What is this organism?

Entamoeba; cyst

What is this organism? Phase?

Entamoeba; trophozoite

What is this organism? Stage?

Gel Immunoprecipitation

What is this test used for?

A type of electromagnetic energy

What is ultraviolet light?

Combination of positive results in a 5 digit numeric code.

What is used to identify the microorganism?

BBL Enterotube II result sheet

What is used to score the results of the Enterotube II test

Gram negative cannot grow due to 7.5% Sodium Chloride.

What kind of bacteria cannot grow in Manitol media?

Selective media for growing gram negative bacteria specifically. Inhibit gram + bacteria growth due to bile salts & dyes.

What kind of media are EMB & Hektoen?

Lentivirus

What kind of virus is HIV?

Litmus milk

What media is this?

N. Broth

What media is this?

N. gelatin

What media is this?

N.A. deep

What media is this?

N.A. slant

What media is this?

Manitol & Sheep blood agar

What media is used to grow gram+ bacteria?

Mannitol salt & Sheep blood agar

What media was used to isolate gram + bacteria?

EMB, Blood agar, Sabouraud

What media were used for the Urine streak?

Fluid Thiogycollate medium (test tube broth)

What medium was used to observe bacteria's reaction to O2?

Enterotube test.

What method is used to differentiate between enteric gram- bacteria?

Kirby-Bauer method

What method is used to test antimicrobial effectiveness?

E. coli Enterobacter Staph. Epidermis Staph. Aureus Strep Pyogenes Candida

What microorganisms were isolated using the Urine streak?

Irreversible damage, cause bacterial death

What occurs after bacteria is exposed to UV-C light for 2 min?

Symptoms appear CD4 T helper cells drop Body becomes susceptible to opportunistic infections

What occurs in the AIDS stage of HIV?

The virus presents no symptoms

What occurs in the latent period of HIV?

Some growth due to DNA polymerase I reversing and repairing damage.

What occurs to bacteria exposed to UV-C light for 30 sec?

No change

What occurs to bacteria with no exposure to UV-C light?

Becomes inactivated when exposed to air. Reduces O2 concentration, increases CO2 concentration

What occurs to the BD gas pack?

Proteus or Salmonella Black color due to production of H2S precipitate

What organisms could be on this Hektoen media and why?

Anaerobic Jar

What other tool is used to see the organism's reaction to O2?

Mitis Salivarius Agar Contains crystal violet and potassium tellurite to inhibit gram (-) bacilli

What plate is this? What does it contain?

-apply wound and skin isolation techniques -wash hands carefully to avoid possible transmission

What precautions should be taken for a patient hospitalized with a superficial mycosis?

-active cases may require strict isolation -masks are required -hands washed -and reduce all potential chances of airborne transmission

What precautions should be taken for a patient hospitalized with a systemic lung mycosis?

B

What proteins would this person have on his/her red blood cells?

Circle

What shape is cocci?

Rod

What shape is rod?

Agglutination test Staph Aureus is positive for agglutination. Staph Epidermis is negative for agglutination

What tests differentiates between Staph Aureus and Staph Epidermis?

Gram negative rod

What type of bacteria is this classified as?

Gram positive cocci

What type of bacteria is this classified as?

Alpha hemolysis

What type of hemolysis is shown?

Beta hemolysis

What type of hemolysis is shown?

Gamma

What type of hemolysis is shown?

Alpha

What type of hemolytic reaction is this?

Beta

What type of hemolytic reaction is this?

Gamma

What type of hemolytic reaction is this?

Anaerobe Grows at the bottom of test tube, where there is no O2.

What type of organism is growing in this test tube?

Microaerophile Growth in the middle of test tube. Does not grow in high O2 or no O2.

What type of organism is growing on this tube?

Sheep Blood Agar

What type of plate is this?

Alpha hemolysis, E. coli or Strep. Pneumoniae

What type of reaction? Possible bacteria this could be?

Beta hemolysis, Streptococcus pyogenes

What type of reaction? Possible bacteria this could be?

-Filobasidielliosis (Cryptococcosis) = tissue biopsy -Athletes foot = skin samples -Tinea capitis = skin samples; hair; use Wood's lamp -Pneumocystis pneumonia = lung biopsy (sputum sample)

What type of specimen should be collected when testing for: -Filobasidielliosis (Cryptococcosis) -Athlete's Foot -Tinea capitis -Pneumocystis pneumonia

Lactose, sucrose, dextrose tests. Middle is negative.

What type of tests and which tube(s) has/have negative result?

Staphylococcus aureus, streptococcus pyogenes, clostridium botulinum and tetani

What types of organisms present beta hemolysis?

when are semi-solid mediums typically used?

When observing the motility of a bacteria because they can easily move and "swim" through the soft medium

Symbiosis

When organisms live in close association with us

What does parfocal mean?

When you have the object in focus with one objective. It will be visible on all objectives with just a minor adjustment from the fine adjustment knob

In the intestine

Where are enteric bacteria commonly found?

A source that infects many people at once

Where does an epidemic usually originate from?

Skin Yes they are pathogenic because they produce beta hemolysis and those bacteria are pathogenic

Where does the bacteria come from that are cultured on these plates? Are the bacteria pathogenic? Why/why not?

E. coli, Enterobacter, Salmonella, Proteus

Which 5 bacteria are identified using the Enterotube?

Proteus

Which NonColiform organism was studied in class?

Lysine Lactose Urea

Which Substances in the Enterotube will be observed for positive results?

UV-C-highest energy, damages bacteria DNA, exposure for a few minutes leads to death.

Which UV light is used to kill bacteria?

Clostridium

Which anaerobe was studied in class?

E.coli and Enterobacter

Which are the two coliform bacteria studied in class?

E. coli Enterobacter Staph. aureus Staph. e.

Which bacteria can be grown on Sabouraud?

Strep. pyogenes

Which bacteria does not grow on Sabouraud?

E.coli uses it, aka photo-reactivation.

Which bacteria forms light repair?

E. coli Enterobacter Staph. aureus Staph. Epidermidis Candida

Which bacteria will grow on the sheep blood agar?

Pneumocystis Carinii Kaposi's sarcoma

Which diseases were discovered in 1981 due to AIDS that our body is normally resistant to?

E.coli & Enterobacter

Which enteric bacteria are non pathogenic i.e. coliforms?

Proteus

Which enteric bacteria are opportunistic pathogens?

Salmonella & Shigella

Which enteric bacteria are true pathogens?

Proteus, Salmonella, Shigella. No color on EMB due to being lactose non fermenters.

Which enteric organisms could be in this EMB media and why?

DNA Photolyase

Which enzyme is used in photo-reactivation?

E. coli

Which facultative anaerobe was studied in class?

Streptococcus Pyogenes

Which gram positive bacteria is responsive to Bacitracin or A disc antibiotic?

Staphylococcus Epidermis Does not ferment Manitol due to 7.4 pH

Which gram positive organism is in this Manitol media and why?

Staph Aureus or Streptococcus Pyogenes Complete hemolysis (beta)

Which gram positive organisms could be on this blood media and why?

E. coli Enterobactor Proteus Salmonella Shigella

Which gram- Enteric bacteria were studied in class?

E.coli has a green metallic sheen Vigorous lactose fermentor

Which gram- enteric bacteria is seen on this EMB agar?

Enterobacter Has a pink color due to lactose fermentation

Which gram- enteric bacteria is seen on this EMB plate?

Micrococcus luteus

Which microaerophilic organism was studied in class?

Streptococcus pneumonia, Enterobacter, E. coli Picture shows Alpha aka partial hemolysis.

Which organism could be growing on this blood agar and why?

Staphylococcus epidermis or Candida Gamma hemolysis-does not hemolyse blood

Which organism is on this blood agar media and why?

Enterobactor or E.coli Pink to orange color due to lactose fermentation.

Which organisms could this be in this Hektoen media and why?

Saroubaud

Which plate is used to grow Candida sp?

Enterobacter

Which specific bacteria is (VP) vouges paskauer positive?

Salmonella, Shigella

Which were the gram+ enteric true pathogens studied in class?

Hydrogen sulfide is produced as a by-product of aerobic respiration in some bacteria because instead of having O2 as the ultimate electron acceptor, they use sulfur

Why is some TSI agar black?

Describe two ways in which smear preparation differs when using a stock brother as opposed to using a stock slant culture.

With broth you must use 5 drops of the broth itself and smear across the whole slide, With the slant you must prepare with 1/2 a drop of water first, add the bacteria and the smear across the whole slide, let both dry

What stains are used in differential blood cells counts?

Wrights or Giemsa stain

These are commonly called fleas and could transmit diseases such as the black plague.

Xenopsylla cheopis

Gram Staining

YOU MAY SEE: -gram negative Bacilli; pink rods -gram negative Cocci; pink cocci -gram positive Cocci; purple cocci -gram positive Bacilli; purple rods -gram stain of mixed culture

Escherichia

_________ is methyl red (MR) postive since it produces drastic amounts of acid, lowering the pH and causing the methyl red to remain red.

define turbid

a cloudiness in a liquid medium caused by cell growth

What is the definition of an infectious disease?

a disease caused by microorganisms that enter the body and multiply in the tissues

Christensen's urea Broth

a media containing urea, among other ingredients and a pH indicator called phenol red, the byproduct of urease can be detected.

What is the Kirby-Bauer agar method?

a method most commonly used to determine the effectiveness of an antibiotic. Antibiotic is added to a plate of a microorganism to create a zone of inhibition. Large zone= sensitive microorganism. Small zone= resistant microorganism. This test is RELATIVE

What is a Durham Tube?

a small glass tube places upside down in the big culture tube

how do antibiotics work?

a specific mechanism targets some vital process of the specific bacteria to cause death

what is UV light used as?

a sterilizing agent

define colony

a visible mound of millions of a bacteria's descendants, all arising from one cell

The tsetse fly: a. can transmit African sleeping sickness b. can transmit malaria c. usually acts as a mechanical vector d. deposits eggs called nits in the victims hair

a. *can transmit African sleeping sickness* b. can transmit malaria c. usually acts as a mechanical vector d. deposits eggs called nits in the victims hair

The antibiotic dispensing unit: a. dispenses antibiotic soaked disks onto plates covered with bacteria b. dispenses five disks at a time c. can be used to perform dilution streak plate techniques d. all of above are true

a. *dispenses antibiotic soaked disks onto plates covered with bacteria* b. dispenses five disks at a time c. can be used to perform dilution streak plate techniques d. all of above are true

Echinococcus granulosis: a. forms hydatid cysts around the body b. has the shape of cocci in clusters c. is treated with antifungal medicine d. is a name I made up

a. *forms hydatid cysts around the body* b. has the shape of cocci in clusters c. is treated with antifungal medicine d. is a name I made up

Mississippi River Valley fever is usually caused by: a. Candida b. Histoplasma c. Taenia d. Mississippi River Mud

a. Candida b. *Histoplasma* c. Taenia d. Mississippi River Mud

A mycotoxicoses could be caused by: a. Pneumocystis carinii b. Coccidiodes immitis c. Aspergillus flavus d. ringworm

a. Pneumocystis carinii b. Coccidiodes immitis c. *Aspergillus flavus* d. ringworm

Which of the following would be a good treatment for roundworm infections: a. Quinine b. metronidazole c. amphotericin B d. mebendazole

a. Quinine b. metronidazole c. amphotericin B d. *mebendazole*

Most intestinal protozoan parasite diseases are best tested by studying: a. blood samples b. stool samples c. throat swab d. urine samples

a. blood samples b. *stool samples* c. throat swab d. urine samples

Infections caused by Trichomonas vaginalis: a. can be treated with quinine derivatives b. are usually sexually transmitted c. are always fatal d. only affect women

a. can be treated with quinine derivatives b. *are usually sexually transmitted* c. are always fatal d. only affect women

Which of the following is not true concerning the nematode life cycle? a. eggs are consumed in contaminated food or water b. eggs hatch into tiny larvae that eventually gain access to the lungs c. larvae are coughed up, swallowed and end up in the intestines where they develop into adults d. the adults are picked up by biological vectors and transmitted to a new host

a. eggs are consumed in contaminated food or water b. eggs hatch into tiny larvae that eventually gain access to the lungs c. larvae are coughed up, swallowed and end up in the intestines where they develop into adults d. *the adults are picked up by biological vectors and transmitted to a new host*

A good example of a mechanical vector would be: a. hard tick b. cockroach c. body louse d. mosquito

a. hard tick b. *cockroach* c. body louse d. mosquito

Chagas disease: a. is a disease I made up b. is common in the desert southwest of the United States c. affects the skin and mucous membranes causing a severe rash d. can be transmitted by the bite of a "kissing" bug

a. is a disease I made up b. is common in the desert southwest of the United States c. affects the skin and mucous membranes causing a severe rash d. *can be transmitted by the bite of a "kissing" bug*

Rat fleas could transmit: a. malaria b. Rocky Mountain spotted fever c. elephantiasis d. scabies e. none of these

a. malaria b. Rocky Mountain spotted fever c. elephantiasis d. scabies e. *none of these*

Cercaria and miracidia are related to: a. protozoans b. yeast c. roundworms d. flukes

a. protozoans b. yeast c. roundworms d. *flukes*

The Platyhelminthes would include all the following except: a. tapeworms b. flatworms c. ringworms d. flukes

a. tapeworms b. flatworms c. *ringworms* d. flukes

Trophozoites refer to: a. the resistant cyst structure made by protozoans b. a class of highly motile protozoans c. deadly flesh eating parasites d. free-living forms of protozoans

a. the resistant cyst structure made by protozoans b. a class of highly motile protozoans c. deadly flesh eating parasites d. *free-living forms of protozoans*

In the Kirby Bauer test, sensitivities to various antibiotics are determined by: a. the streak plate technique b. using unpaid volunteers c. using a metric stick d. growing the bacteria in broth tubes

a. the streak plate technique b. using unpaid volunteers c. *using a metric stick* d. growing the bacteria in broth tubes

With the hard ticks: a. they usually burrow into the skin and cause intense itching b they can exhibit transovarian transmission c. they have six legs, jaws, and antennae d. they only invade the pubic areas

a. they usually burrow into the skin and cause intense itching b *they can exhibit transovarian transmission* c. they have six legs, jaws, and antennae d. they only invade the pubic areas

Phenylalanine Deamination "Phenylaline Agar" - purpose?

ability of an organim to deaminate phenylalanine resulting in the production of phenylpyruvic acid and ammonia (NH3) - enzyme used is phenylalanase

Carbohydrate Fermentation Test - purpose?

ability of an organism to ferment a specific carbohydrate with or without the production of gas

Tryptophan Hydrolysis "Indole Test" - purpose?

ability of organism to split indole from the amino acid tryptophan using tryptophanase

Voges-Proskauer "MR-VP Broth" - detects what product?

acetoin

Decolorizer

acetone/alcohol 5 seconds

What is used in the decolorzation step of an acid fast stain?

acid alcohol

Which cells retain the primary stain in an acid fast stain?

acid-fast bacteria retain the carbolfuchsin dye because their cell walls contain mycolic acid (lipid) which retains the dye

Is yogurt acidic or alkaline

acidic, makes lactic acid

Culturing

act of growing

eosinophil

active in worm or fungal infections allergies & inflammatory response.

Voges-Proskauer -"MR-VP Broth" - once incubated what needs to happen?

add 18 drops each of Barritt's solution A and B

Methyl Red Test "MR-VP Broth" - After incubation what do you do?

add 3-4 drops of Methyl Red Reagent

rh typing

add anti-D to the samples. if there is agglutination within 2 minutes this is rh positive. no agglutination, rh negative. after the first pregnancy if the mother is rh negative, anti-rh antibodies can attack the fetus

Epidemic diseases

affect many individuals in population in short time

What does an iris diaphragm do?

affects contrast between the light and dark parts of the specimen,

When is the mordant (gram's iodine) added in gram staining?

after Crystal Violet is applied

Tryptophan Hydrolysis "Indole Test" - procedure?

after incubation is complete add 10 drops of Kovacs Reagent

what is a solid medium called?

agar

Motility Test - contents?

agar and TTC

PEA agar with blood

agar only

blood agar

agar only

chocolate agar

agar only

TSA agar

agar only (yellow)

What enzyme is linked with the antibodies?

alkaline phosphatase

define streaming

all objects "flow" with the movement of water

semisolid medium (deeps)

allow bacteria to grow down in the medium where there is less oxygen than at the surface. deeps allow for the detection of motility by growth away from initial inoculation site.

differential media

allow colonies to be different colors based off of some characteristics

how would you explain the purpose of blood in the blood agar medium

allows detection of hemolysis reaction

Differential medium

allows for growth of all bacteria while differentiating them from one another

Selective medium

allows the growth of certain bacteria while preventing growth of other bacteria

what would a green reaction code for

alpha

Viridans & Strep. pneumoniae

alpha hemolytic (incomplete/ greenish color)

If milk is pasteurized, what causes it to spoil in such a short time?

although pasteurized, there are still live bacteria present in the sample

How does oxygen affect the growth of bacteria?

amount of oxygen present in an environment can be vital to growth of bacteria.

What is catalase?

an enzyme breaks down H2O2

What is catalase?

an extracellular enzyme produced by some bacteria in order to detoxify Hydrogen Peroxide (H2O2).

DNAse is:

an extracellular enzyme that is produced within the cell and secreted outside of the cell wall

agar

an extract from marine red algae

vector

an organism, a biting insect or tick, transmits a disease or parasite from one animal or plant to another.

What type of environment will allow nitratase be used?

anaerobic conditions so the organism can derives its oxygen from nitrate

B anthracis causes what disease?

anthrax

B cells produce what in the body?

antibodies

known what molecules are on the latex beads that specifically detect s. aureus

antibody a

type AB blood has

antigen A & B and no antibodies agglutination in both A & B

type a blood has

antigen A, and B antibodies agglutination occurs when anti-A serum is added

type b blood has

antigen B, and A antibodies if agglutination occurs when anti-B serum is added

What is a chemotherapeutic agent/antibiotic?

antimicrobial compounds that can be safely absorbed or taken internally

Antigen

any substance that produces an immune response in the body.

What is ARA?

arabinose -promoter that activates GFP and Amp resistance genes

what is the plasmid in the Biorad kit

arabinose used to turn "on" the glow portion of this plasmid. this plasmid contains the ampicillin resistance cassette and a gene for a green fluorescence protein (GFP). once expressed the GFP will fluoresce under UV light. If bacteria have taken up the plasmid, the colonies should fluoresce green under UV light

Microscope diaphragm

attached to bottom of stage allows control of amount of light that will pass through the slide.

What is the most effective form of sterilization?

autoclaving

What is aseptic technique?

avoiding contaminants, preventing infection

What are fermenters?

bacteria can utilize glucose in the presence or absence of O2

What is a vegetative cell?

bacteria cells that are metabolizing and reproducing rapidly

halophile

bacteria that grow better in environments with high salt concentrations

MacConkey Agar (MAC) - differential

bacterial colonies that can ferment lactose turn the medium any variation of red. the red color is due to the pH indicators response to acidic environment created by fermenting lactose. organisms that do not ferment lactose do not cause a color change

If the chemical is killing the bacteria it is said to be....

bactericidal

If the chemical is inhibiting the bacteria growth it is said to be...

bacteriostatic

how is the relationship established between the organism and the legume

bacterium binds to root hairs of the plant, sends out an infection thread that invades the deeper root tissue and forms a small tumor-like structure called a root nodule

basophil

bag of purple balls

What will be in high numbers if the patient has an active infection?

band - neutrophils

Immature neutrophils...AKA?

bands

basis of blood grouping

based on the surface antigens

Is ammonia acidic or basic?

basic

What are the least abundant WBCs in normal blood?

basophils

why are spores not easily stained, and why is heat applied?

because of their hardy nature, therefore, a heat must be applied so the stain can penetrate the spore.

why is detecting pathogens in water difficult

because they are present in such low numbers by the time they are detected it is usually too late to prevent an outbreak of disease.

why are hypertonic environments (higher osmotic pressure) often less tolerated by bacteria?

because water flows out of the cell, causing the cell to dry out.

no blood on the plate what reaction is that.

beta

MAC agar contents

bile salts neutral red lactose

MAC agar selective agent

bile salts - support gram negative

what are the detectable differences between bacteria?

biochemical in nature, and reflect different evolutionary paths. example one may use lactose but not glucose or may use glucose but not fructose. subtle differences are bacteria may use lactose as an energy source accomplished through different pathways, utilizing different sets of enzymes during the process.

Students do various biochemical tests and learn that bacteria has enzymes that can breakdown

biochemicals ( substrate ) and produce visible reactions that help to identify microrganisms

Hydrogen Sulfide Production "Triple Sugar Iron Agar" - positive test?

black precipitate along stab line

How is the Ferrous sulfide seen in the medium?

black precipitate in the medium

Stphylococcus aureus

boils and pimples

T cells are produced where in the body?

bone marrow

Clostridium and Bacillus cause what deadly diseases of humans?

botulism and tetanus

BHI

brain heart infusion agar

What is hydrolysis?

breakdown of starch

catalase enzyme

breaks down hydrogen peroxide to water and oxygen. needed for an enzyme to exist in the presence of oxygen.

if an organism is a fermenter what would you see in a macconkey agar plate

bright pink

eosinophil

bright red background

What is the pH indicator for Citrate test?

bromthymol blue

The methyl red (MR) and Voges-Proskauer (VP) tests are read from a single inoculated tube of MR-VP broth. After 24-48 hours of incubation the MR-VP

broth is split into two tubes. One tube is used for the MR test; the other is used for the VP test.

thioglycolate broth

broth used for oxygen requirement of bacterial growth

Voges-Proskauer "MR-VP Broth" - negative result?

brown color develops

Carbohydrate Fermentation Test - gas production positive test?

bubble forms in Durham tube

How is the number of viable bacteria determined?

by calculating the number of colony forming units per milliliter of sample. this is done by diluting the initial sample through serial dilutions.

how are antibiotics produced?

by microorganisms like fungi. In human and healthcare they are derived from microorganisms or are synthetically produced

facultative anaerobes

can grow with or without oxygen, but usually grow better in presence of oxygen

Describe a Capsular Stain (capsule)

capsule shape with a halo around it

What family of organic compounds does agar belong to?

carbohydrate

carbohydrate fermentation test

carbohydrates can be broken down by fermentation resulting in acid production. This test is used to determine the ability of an organism to ferment a specific carbohydrate with or without the production of gas, positive result=yellow color change with or without gas production. negative= no color change

What is the primary stain of an acid fast stain?

carbolfuchsin dye (red)

if a microbe can degrade hydrogen proxied what is the name of the enzyme that must be able to produce

catalase

What is the biochemical reaction for catalase test?

catalase H2O2 -------------------> 2H2O2 + O2

Negative MR (Methyl Red) BROTH test : Klebsiella and Enterobacter produce more neutral products from glucose (e.g. ethyl alcohol, acetyl methyl carbinol)

causing the pH to rise above 6.2. At this pH, methyl red indicator is a YELLOW COLOR (a).

what influences growth of bacteria?

chemical and physical factors. bacteria differ in in requirements for oxygen, temperature and osmotic pressure

growth medium (plural, media)

chemically defined or may contain partially digested proteins (peptones) or even more complex substances such as bovine red blood cells

define antiseptics

chemicals that are used on the skin or wounds to decrease the number of microorganisms

antibiotics

chemicals that have the ability to kill bacteria. often produced by microorganisms like fungi and are used as a defense mechanism

define disinfectants

chemicals used on inanimate objects

antiseptics

chemicals used on living tissue to kill microbes (ex/ mouthwash)

disinfectants

chemicals used on the surface of inanimate objects to kill microbes (ex/bleach)

What are spores resistant to?

chemicals, heating and basically all harsh conditions

What does chromagen do in the ELISA test?

chromagen will react with the enzyme and break down to show the blue color

zone of inhibition

circle around the disk where the antibiotic noticeably affect the cell viability. this is the area in which the effectiveness of antibiotic on bacterium is determined.

What shape is coccus/cocci?

circular/round dot like

What is the biochemical reaction for Citrate?

citrate ----------------> alkaline end product

Describe an Neutrophil cell

clear granules

DNase agar - postive test?

clear halo will appear around the areas where the DNase-producing organism has grown

How should the iris diaphragm be positioned fore low-power objective?

closed

What is the function of a platelet cell?

clotting

What is the biochemical reaction for coagulase test?

coagulase fibrinogen-----------------------> fibrin clot

What tests are needed to confirm Staph aureus?

coagulase, blood agar, and catalase.

psychrophile

cold - loving; optimum growth at 15 degree Celsius - 20 degree Celsius. commonly isolated from cold waters such as the bottom of the ocean. many grow well at temps of 0 degree Celsius - 4 degree Celsius (even below freezing in the presence of salt)

Mannitol Salt Agar (MSA)

common medium used for isolation of some pathogenic Gram positive organisms.

How does an epidemiologist decide wheather an epidemic really exists or not?

comparing the numbers of new cases with previous records

What is a phenol coefficient?

comparison of the killing action of the compund to that of phenol under the same conditions

beta

complete

What is beta hemolysis

complete hemolysis, is a complete lysis of red cells in the media around and under the colonies: the area appears lightened (yellow) and transparent.

state four factors that influence the size of inhibition

concentration of antimicrobial solubly

explain the pGLO positive gene regulation system

controlling the green fluorescent protein (GFP). the sugar arabinose turns the gfp gene on. also contained in the pGLO plasmid is the araC gene. When arabinose is taken up, it interacts with araC protein, which is bound to the DNA. This interaction causes araC to change shape, which helps RNA polymerase to bind to the Pbad promotor and transcribe the GFP gene. This enables the transformed cells to fluoresce green under ultraviolet light when arabinose is added to the nutrient agar.

complex media

culturing medium that contains nutrients released by partial digestion of yeast, beef, soy, or other proteins; thus, exact chemical composition is unknown

What's a BEA positive test?

dark brown

what is the appearance of fecal coliforms on mLES agar

dark colonies with a metallic sheen

What is the critical step of gram staining?

decolorizer is critical and the timing

this is mannitol salt agar, the s aureus, pathogenic, are able to ferment mannitol sugar and change color around bacterial colonies to a yellow halo

describe what is happening here and what medium

What's the purpose of Bile Esculin Azide agar test for?

detecting Enterococcus faecalis

multiple tube technique

detects coliforms involves two tests: presumptive test and confirmation test

oxidase test

detects presence of cytochrome oxidase involved in reduction of oxygen at end of E.T.C. this enzyme reacts with kovacs reagent creating a purple edging effect.= positive. negative = no edging effect

H2S production test

detects the enzyme that liberates hydrogen sulfide from a sulfur containing organic compound. demonstrates the production of hydrogen sulfide in Iron Peptone Agar. positive= growth with black precipitate in the media. negative= growth with no black precipitate

phenylalanine deamination test

detects the presence of phenylalanine deaminase which knocks off amine groups of molecules to form an acidic product. this test utilizes the amino acid phenylalanine that is converted to phenylpyruvic acid which is detected via a reaction with reagent FeCl3. positive= dark green color on slant. negative = no color change

What is the reason for coagulase test?

determine the ability of an organism to clot plasma by the action of the enzyme, coagulase

Hydrogen Sulfide Production "Triple Sugar Iron Agar" - purpose?

determine the ability of an organism to hydrolyze the sulfur containing cysteine to produce pyruvic acid, ammonia and hydrogen sulfide. -enzyme used: cysteine desufurase

What can B. cereus cause?

diarrhea

How can gram positive and gram negative bacteria be differentiated?

differences in their peptidoglycan layer

What property of bacterial cell structure makes the Gram Stain possible?

different in cell wall thicknesses

Is acid-fast stain simple or differential stain?

differential

Mannitol salt agar does what?

differential & selective media. differentiates pathogenic from not pathogenic.

DNase agar - purpose?

differential medium that tests the ability of an organism to produce an exoenzyme called DNase that hydrolyzes DNA

What is Blood Agar differential or selective?

differential, differentiates between those bacteria able to make the enzyme, hemolysis and those that cannot

differential stain

differentiates between two types of bacteria. visualizes the bacteria. example: gram stain

describe the Spread plate method

diltuing a bacteria sample before adding it to the agar so bacteria can be visualized easier and be counted more easily

describe the pour plate method

diluted cells are mixed with molten agar when it is still warm so that when it hardens into a solid, the microorganisms can grow inside and on top of the medium

Describe the streak plate method

diluting a bacteria enough on a certain area of an agar plate and spreading the bacteria out enough so that colonies can form

What is the dilution factor of the following dilutions: 1/100 X 1/10 X 1/10 X 1/10 X 1/10

dilution factor = 100,000

what did scientists start to notice as a lack of aseptic techniques?

directly related to incidences of fever and infection after childbirth, because students at the hospital would commonly go from the autopsy room to assist with delivery without washing hands.

Zoonoses

diseases of animals that can spread to humans

contagious (communicable) diseases

diseases transmitted person to person

What is the principle of technique for the Kirby-Bauer?

disk diffusion

hemolysis

done on SBA plates and recorded as alpha (partial sometimes with a greenish appearance), beta (complete) or gamma (none)

why do bacteria tolerate hypotonic environments?

due to the rigidity of the cell wall and the peptidoglycan layer

how do chemicals affect bacteria?

each chemical has a microbe that works most effectively against an environment which it works best in.

what does aseptic technique achieve?

enables pure cultures to be produced by keeping the culture free of contaminants. It also protects the microbiologist and the environment from the culture.

Coliform are what member of bateria?

enterobacteraciae (gram - / lactose fermenters)

what does ELISA stand for

enzyme-linked immunosorbent assay

hemolysins

enzymes that lyse host cells, especially red blood cells.

What increases with parasitic infections?

eosinophils

define Brownian motility

erratic vibration of a particle cause by the unequal collision of water molecules against it

Are red blood cells prokaryotic or eukaryotic?

eukaryotic

chemically defined media

exact chemical composition

what happens if you overheat the slide when heat-fixing?

excessive heat can destroy cells and make it impossible to identify

What are amylase, proteases and ureases all catagorized as?

exoenzymes

Gram's Iodine - mordant

facilitates further interactions between primary dye and cell wall. crystallizes dye that has penetrated the microbe

Vector of malaria

female anopheles mosquito

WALK AWAY PANEL if the sugar turns yellow?

ferments sugar (sucrose)

Hydrogen Sulfide Production "Triple Sugar Iron Agar" - contents?

ferrous salts that react with H2S to produce an insoluble black precipitate "sulfate"

know what molecules are on the surface of the bacteria that are detected by the latex beads

fibrinogen binds to clumping factor B

Why is serial diluting performed?

for plate counting and several techniques for isolating and maintaining pure cultures

Methyl Red Test "MR-VP Broth" - enzyme that breaks down formic acid?

formic hydrogen lyase

How does UV light kill organisms?

forms thymine dimers and prevents normal DNA replication

What is the function of a Eosinophil?

found in the blood due to parasites and allergies

sterile

free from bacteria or other living microorganisms; totally clean

enzymes

function to lower the activation energy of reactions that would otherwise be too costly in energy or too slow for successful growth of the bacteria. many bacteria need these enzymes to breakdown certain materials in the environment for energy requirements. if bacteria lost function of some enzymes they would no longer be able to survive in certain environments.

what does C. perfringens cause?

gas gangrene and perfringens food poisoning

Gelatin Liquéfaction test - positive test?

gelatin is liquefied

Gelatin Liquéfaction test - negative test?

gelatin is solid

what are the two parts of a scientific name?

genus and species or specific epithet

What is the biochemical reaction for OF Glucose Oxidizer?

glucose + O2 -------------------> acid end product

What is the biochemical reaction of OF Glucose Fermenters?

glucose --------------------> acid end product

What is the biochemical reaction for Methyl Red?

glucose ------------------>acid end product

What is the biochemical reaction for Voges-Proskauer?

glucose-----------------> neutral end product

ampicillinase

goes out into the environment and destroys the ampicillin in the media

A plate with a RESISTANT (no zone) Vancomycin would be gram + or gram -?

gram -

PLATE a Maconkey plate that has turned green, name the organism

gram -, oxidase +

EMB agar - selective

gram negative

BA - selective for?

gram positive

macconkey agar is inhibitory for gram negative and

gram positive

MSA agar - selects for?

gram positive because gram negative cannot grow in presences of salt

What does Staphylococcus look like

gram positive purple cocci in groups

What are the characteristics differential and selective media use to distinguish between various types of bacteria?

gram reaction, sugar fermentation, and tolerance of salt

what is the first and most important step in identifying bacteria?

gram staining

On a blood smear which is the Erythrocytes RBC?

gray color (background cells)

What color are spores when dyed?

green

Phenylalanine Deamination "Phenylaline Agar" - positive test?

green color develops

PGLO

green florescent Protein-GFP AMP function?- kill bacteria and select resistant population ARA function?- An inducer that causes bacteria to glow. which media glowed? LB/ARA/AMP

What is GFP?

green fluorescent protein -used an indicator of successful transformation and gene transcription

obligate aerobes

grow in presence of oxygen

obligate anaerobes

grow only in the total absence of oxygen

what does the Centers for Disease Control and Prevention state?

hand washing is the single most important procedure for preventing nosocomial infections (infections acquired in a hospital)

What does the coagulase enzyme do?

hardens a protective fibrin against phagocytes.

What does a condenser do on a compound microscope?

has a lens that gathers light and helps resolution.

thermophile

heat - loving; optimum growth at 50 degree Celsius - 60 degree Celsius (sometimes even higher). found in hot springs such as in Yellowstone

How does heat kill all organisms?

heat denatures their enzymes and inactivates them

what is a physical property of bacterial species?

heat sensitivity

What will allow the DNA "plasmid" to enter the E.coli?

heat shock "opens pores"

What are the two things to insure uptake of the gene

heat shock and calcium choloride

What are the ingredients used to make yogurt

heat, milk, cup, spoon, non-fat dry milk, culture, aluminum foil, incubator, glass rod for stirring

pasteurization

heating at 63 degree Celsius for 30 minutes or 72 degree Celsius for 15 minutes

What is the purpose of acid fast?

high cell wall content and to identify Mycobacterium and Nocardia

MSA - selective

high salt concentration of this medium inhibits the growth of most other organisms (halotolerant)

If positive test after 24 hours with Snyder test?

high susceptibility to dental caries

What is the effect of temperatures on bacterial growth?

high temperatures will kill bacteria while cold temperatures will inhibit their growth

the x40 ocular lens is also called?

high-dry objective

What shape does the monocyte nucleus have?

horseshoe shaped

dry heat

hot air, such as a hot oven. flaming inoculating loops is an example of a sterilization technique that uses dry heat.

carriers

human reservoirs that are infected without having symptoms

reservoirs

humans or animals that are sources of infection.

Station 1 Gelatin test Principle

if bacteria produces Gelatinase as they grow, they will breakdown the substrate which is gelatin.

Lymphocyte

immune reactions to foreign matter 20-35% 2nd place

moist heat

implies that steam is involved. example of sterilization techniques that uses moist heat are: boiling, autoclaving, and pasteurization. denatures proteins.

After medium is prepared how is it sterilized before use?

in a steam autoclave

why does water flow in or out of the cells?

in an attempt to make solute concentrations inside and outside the cell equal

Microbial biochemical characteristics are brought about by enzymatic reactions that are a part of specific metabolic pathways .The tests used

in this section were selected to illustrate process that leads to identification.

How does gram stain differentiate cells?

in two classes gram positive or gram negative based on the structure of the cell wall

fomites

inanimate objects such as surfaces, bedding, towels, pandemics drinking vessels, or needles.

polyenes, anti-fungal agent

includes amphtericin and Nystatin. binds to ergosterol component, results in a leaky membrane. it is very toxic, limited to use only in life threatening conditions. (Hint: POLY-many - combined - binding)

azoles, anti-fungal agent

includes clotrimazole, miconazole. inhibit ergosterol synthesis, without ergosterol the cell's membrane does not remain intact and the fungal cell dies. (Hint: a-against-inhibits ergosterol)

How are H2O2 molecules formed in a cell?

incomplete reduction

what is the result of higher solute concentration?

increase of osmotic pressure

urea hydrolysis

indicator= phenol red positive result color= pink positive organism= proteus vulgaris enzyme used= urease

how does temperature affect the growth of bacteria?

individual bacterial species usually grow over a range of temperatures. they have a temperature at which they grow best.

MacConkey Agar (MAC) - selective

inhibit growth of Gram-positive organisms allow for gram - negative to grow

How do you label test tubes and petri dishes?

initials-section # JHW-32 date 10-4-10 Medium BHA Organism/Bacteria E. coli

mechanical transmission

insect carries fecal pathogens on its feet and then walks on food

Urea Hydrolysis "Urea Broth" - positive test?

intense pink color

repair mechanism

involves enzymes which physically remove and replace the structural changes caused by radiant energy

Secondary stain

iodine/Gram -/1min/ Mordant

What do we use on a microscope to brighten an image?

iris diaphragm

Positive MR (Methyl Red- Voges-Proskauer) BROTH test : E. coli is one of the bacteria that produces acids, causing the pH to DROP below 4.4. When the reagent methyl red

is added to this acidic broth it will be cherry red

how can you determine if a microbe can degrade hydrogen proxied in a lab

it has to bubble

What does the term pasteurization mean?

it is a heating process designed to preserve the properties of the liquid but sufficient to kill any pathogens

Coarse adjustment knob?

it is used for bringing objects into initial focus with lower power objectives, never be used with high dry or oil

bacteriocidal vs. bacteriostatic

kills bacteris; temporarily inhibits bacterial growth

what sugar is the differential medium used to detect the presence of fecal coliforms

lactose

MacConky agar uses what biochemical reaction

lactose fermentation

macondky entric bacteria based on what biochemical reaction

lactose fermentation

What does strong fermentation of sugar produce?

large quantities of acid

Describe the gram positive peptidoglycan layer.

larger and more highly cross-linked containing small pores

Describe a yeast

larger than all other

What is another way in which media can be prepared?

liquid medium (media: broth), after incubation, liquid media will appear turbid due to bacterial growth this type of media is helpful when the bacteria need to be diluted.

Station 1 Gelatin test Positive result:

liquid, means that the bacteria produced Gelatinase as they grew, and broke down the substrate gelatin

what kind of bacteria are Clostrdium and Bacillus?

long gram-positive rods

the x10 ocular lens is also called?

low-power objective

give an example of a media that is both selective and differential

macconkey agar selects for gram negative and differentiates for lactose fermentation

bacterial genome

made up of the bacterial chromosome plus any plasmids that the organism may contain.

What is the function of the phospahte buffer for the urease test?

maintains ph

What is used to stain spores?

malachite green

Which dye is used to dye the actual endospore?

malachite green

MSA differential agent

mannitol and phenol red

What is the biochemical reaction for Mannitol Salt Agar?

mannitol----------------->acid end product

motility test

many organisms are motile because they possess flagella. flagella come in a variety of arrangements and can allow the organism to move very quickly in some cases. motility agar deeps are sued to test for motility. positive= fanning out away from the stab line. this can also make the tube look cloudy. negative= no fanning out from the stab line.

Why must solid and liquid samples be diluted prior to testing?

many samples are heavily contaminated with bacteria and must be diluted to provide an opportunity to visibly count colonies

what happens to gram positive cells if ethanol is left on for too long?

may decolorize causing misidentification of the bacteria

What happens if you plate the sample undiluted?

may have too many colonies to count

what happens when agar is added to growth medium?

medium becomes a solid

autoclaving

medium is heated to 121 degree Celsius at 15 psi for 15 minutes. this is sufficient to kill all microbes, including spores

Carbohydrate Fermentation Test - acid production negative test?

medium remains red

Carbohydrate Fermentation Test - acid production positive test?

medium turns yellow

if an organisms is e.coli what will you see on the EMB plate

metallic green

aseptic technique

method for transferring cultures from one medium to another without contamination

What is meant by binomial system of nomenclature?

method of scientific name-combination of genus and species, identify a species

What is the counter stain in an acid fast stain?

methylene blue (blue..obvi)

define ubiquitous

microorganisms seem to be present everywhere

How would you distinguish among the four classes of protozoans?

microscopic observation of the shape, size, number and type of nuclei, and type of motility

mesophilic

middle - loving; optimum growth at 30 degree Celsius - 37 degree Celsius. Pathogens of humans, grow best at body temperatures.

What kind of plate of used for the Casein hydrolysis test?

milk plate: nutrient broth with a skim milk additive

If positive test after 48 hours with Snyder test?

moderate susceptibility to dental caries

Why is moist heat more effective than dry heat as a sterilizing method?

moist heat penetrates material better than dry heat

What is the largest WBC?

monocytes

coliforms

most commonly used indicator. aerboic or facultatively anaerobic gram negative rods. do not form endospores and they ferment lactose which leads to acid and gas production

MPN

most probable number or MPN method. determines the number of coliforms in a sample

coarse and fine focus knobs

moves stage up towards objective lens or moves objective lens down to stage in order to focus image

Calculate the total magnification

multiply the magnification power of the lens, by the eyepiece lens 10x

Name Two genera that contain species that are acid-fast.

mycobacterium & Nocardia

Mycoplasma

neg for Gram Rxn-no cell wall

what general catagory of stains does capsule stain belong to?

negative stain

Color of pH indicator "Bromcresol green"

neutral -green acidic -yellow

MAC agar differential agents

neutral red lactose

What is the most abundant WBC?

neutrophils

type O blood has

no antigens, antibody A & B no agglutination

catalase: Negative result, Positive result, media used & reageant added

no bubble formation, bubble formation (oxygen-release), nutrient agar slant, hydrogen peroxide

Carbohydrate Fermentation Test - gas production negative test?

no bubble forms

Protozoa characteristics

no cell wall, organelles, asexual forms

Citrate Utilization - negative result?

no color change

Oxidase Test - negative test?

no color change

Decarboxylation Test - negative test?

no color change **may turn yellow if a glucose fermenter

Nitrate Reductase Test - negative test?

no color change - do confimation test by adding a small pinch of zinc powder

Urea Hydrolysis "Urea Broth" - negative test?

no color change -continue to incubate for 7 days for slow urease producers

y-hemolytic

no damage

What color do neutrophils stain?

no intense color

Hydrogen Sulfide Production "Triple Sugar Iron Agar" - negative test?

no precipitate

Station 1 Gelatin test Reagents added after incubation

no, but take tubes out and put into cold room for 20 minutes.

Do simple stains have a decolorization step?

no.

If urine samples were tested, should the presence of a few hundred bacteria indicate infection?

no; all normally collected urine samples have a few contaminants from the urethra

Will the use of serial dilutions indicate the presence of pathogens in a test sample?

no; serial dilutions only indicate the number of bacteria in a sample

Which cells do not retain the primary stain in an acid fast stain and what color do they end up?

non-acid-fast cells, blue

What kind of radiation is UV?

non-ionizing radiation

gamma

none

NA

nutrient agar

What is a common agar used in lab?

nutrient agar (NA), made of peptone, beef extract, sodium chloride, agar and distilled water.

What does paracentric mean?

objects place in exact center of the viewing field using one objective will still be centered after changing to another objective.

Is Clostridium an obligate or an aerobic anaerobe?

obligate anaerobe while most species of bacillus are aerobic

How is total magnification found? (mathematically)

ocular lens power (constant) x objective lens power

the x100 ocular lens is also called?

oil-immersion objective

Why do older bacteria not give reliable results when doing gram stain (+)?

older gram stain (+) cells bleach easier than younger cells and can stain pink which makes them appear gram (-)

Simple stain

one dye and dyes the cells

selective

only allow some organisms to grow; some inhibit growth of certain bacterial species

How many times is the fine adjustment know to be turned?

only slightly either direction

How do you know e.coli has been transformed if it grows on the medium treated with ampicillin

only transformed e.coli will contain the beta lactamase

how should the iris diaphragm be positioned for oil immersion objective?

opened

How do you calculate total magnification?

optic x ocular

index case

original source case

Decarboxylation Test - what must be done after inoculation?

overlay of mineral oil to seal the medium from external oxygen and promote fermentation

Selective ingredient for BEA test include:

oxgall (inhibits gram +) Sodium azide (inhibits gram -)

PLATE proteus is?

oxidase -, gram - rod

What reagent is used for the oxidase test?

oxidase reagent vial

What is the function of a Red Blood Cell?

oxygen and carbon dioxide transportation

What is necessary in an experiment to gain fluorescence?

pGLO gene, arabinose, ampicillin

What is the vector used in the transformation lab?

pGLO plasmid

MAC agar - purpose of neutral red

pH indicator low pH - red/purple/pink colonies neutral - colorless

MSA - purpose of phenol red

pH indicator low pH - yellow neutral pH - orangish red high pH - pinkish red

Bromothymol blue

pH indicator used in citrate test

disk-diffusion method

paper disks impregnated with various antibiotics are placed on a lawn of bacteria. antibiotic will diffuse into the media during incubation

What are peptones?

partial breakdown products of proteins

What does weak fermentation of sugar produce?

partial masking of the alkaline end products of peptone breakdown

alpha

partial, sometimes with a greenish appearance

Name the enzyme that help helps bacteria to develop resistance against penicilin?

penicilinase/beta-lactimase

What is the biochemical reaction for peptone breakdown?

peptone +O2 -------------------------->alkaline end-product

What does the carbohydrate fermentation media contain?

peptone- to support growth of the bacteria. carbohydrate- to test the bacteria. phenol red- pH indicator that changes to yellow to indicate acid production and a Durham tube- to collect gas that is formed.

What is the biochemical reaction for Peptones?

peptones + O2 --------------------> alkaline end product

What is the biochemical reaction for OF Glucose Peptones?

peptones + O2-------------> alkaline end products

What is a common way of preparing media?

petri plates, contain a solid media and provide a large surface area for isolating and observing colonies with the naked eye

What is the function of a Macrophage (lung)?

phagocytosis

What is the function of a Neutrophil?

phagocytosis

What is the pH indicator for Mannitol Salt Agar test?

phenol red

What is the pH indicator of TSI agar?

phenol red

What is the ph indicator for urease test?

phenol red

what is the pH indicator and percent salt solution used in mannitol agar

phenol red, 7.5% sodium chloride

what is fastidious?

picky about nutritional requirements

Tryptophan Hydrolysis "Indole Test" - positive test?

pink layer forms on surface of the medium

Mannitol Salt Agar

pinkish agar. positive - yellow surrounding colonies negative - no change in color around colonies presumptive test for s. aureus

What does "p" mean in transformation Experiment?

plasmid

high copy number

plasmid may have up to 300 copies of itself in any one bacterial cell.

pGLO

plasmid that contains an ampicillin resistance gene codes for exoenzyme ampicillinase.

Trichinella spiralis

plate Ham/Eggs

How many colonies are best for counting?

plates containing between 30 and 300 colonies

streptococcus pneumoniae

pneumonia

neutrophils...AKA?

polymorphonuclear neutrophils "PMNs" 2 to 5 lobes

What is a capsule?

polysaccharide-containing layer outside of a bacterial cell that can increase the bacteria's virulence, survival of desiccation and attachment to surfaces.

What is UV light limited by?

poor penetrating power

EMBA- eosin methylene blue agar

positive color result= green & pink indicator= eosin and methylene blue Positive organism= E.coli Carbon Source= Lactose

WALK AWAY PANEL urea is pink what does this mean?

positive for urea

be able to recognize positive and negative latex agglutination reactions

positive is the left blue circle showing clumps negative is the right blue circle, no clumbs

Starch Hydrolysis

positive organism= bacillus indicator= iodine clearing means= amylase present enzyme used= amylase negative organism = E.coli

MSA test

positive= growth of bacteria with a color change of media to yellow. negative= growth of bacteria without color change or no growth of bacteria

MAC test

positive= growth of bacteria with color change of media to bright pink. negative= growth of bacteria without color change or no growth of bacteria

capnophillic organisms

prefer higher concentrations of CO2 (5% - 10%)

endemic diseases

present in population all the time at low levels

Coagulase Test

presumptive for staph. aureus.

What are the 2 dyes used in a gram stain

primary dye (Crystal Violet) and a counter stain (Safranin)

With such a high count of bacteria in spoiled meat or milk, may we assume that the sample contains pathogens? Explain.

probably yes- the greater the contamination, the more likely the presence of pathogens

How do aerobic cells protect themselves from the effects of H2O2?

produce catalase

What is the function of a lymphocyte?

produces antibodies

OXIDASE Theory : A positive oxidase reaction reflects the ability of a microorganism to oxidize certain aromatic amines, such as tetramethyl-p-phenylene diamine (TPD),

producing colored end products. This is due to the activity of cytochrome oxidase in the presence of atmospheric oxygen.

Is Trypanosoma cruzi prokaryotic or eukaryotic?

prokaryotic

what are the proportions of osmotic pressure?

proportional to number of particles dissolved (solutes; often salt or sugar) in a certain volume of solvent (usually water)

Casease enzyme:

protease exoenzyme

Capsules

protective polysaccharide layer

Decarboxylation Test - positive test?

purple

What color do basophils stain?

purple

What color will gram positive bacteria be?

purple

On a blood smear which is the Lymphocytes?

purple color

Slide is of Cerebral Spinal Fluid and is a Gram + cocci. Describe what it looks like. What organism is it?`

purple round shape; strep pneumo

gram positive cocci

purple round shaped

define true motility

purposeful and directional swimming

What is produced when UV radiation is absorbs by DNA?

pyrimidine dimers - which interferes with DNA replication and transcription

What medium is used to test for coagulase

rabbit or human plasma positive is confirmatory for s. aureus

understand the basis for latex agglutination reactions

rapid confirmatory tests for s. aureus checking for protein A and coagulase

basophil

rarest less than 0.5% inflammatory events

Nitrate Reductase Test - positive test?

red color (nitrate reduced to nitrite)

Methyl Red Test "MR-VP Broth" - positive result?

red color develops

Nitrate Reductase Test - negative confirmation test?

red color- nitrate reduced by zinc, not the organism (confirms 1st negative)

Describe an Eosinophil cell

red granules

gram negative rod

red rod shaped

Slide is of Cerebral Spinal Fluid and is a Gram - cocci. Describe what it looks like. What organism is it?`

red round shape; Neisseria Meningitis

Station 14 TSIA Second tube

red slant/red butt - no fermentation. Peptone catabolized aerobically and anaerobically with alkaline products.

What color will gram negative bacteria be?

red/pink

When is phenol red red and when is it yellow?

red= pH above 7 yellow= pH below 7

osmotic pressure

refers to pressure required to prevent flow of water by osmosis.

dark repair

repair mechanism that does not require visible light.

microaerophiles

require low concentrations of oxygen (2% - 10%) for growth. higher oxygen concentrations will inhibit their growth and they cannot grow in the total absence of oxygen

WALK AWAY PANEL antibiotics are cloudy this means?

resistant - not a good drug to use

how are bacteria classified to an antibiotic based on the diameter of the zone of inhibition?

resistant, intermediate, susceptible

What are endospores?

resistive structures that resists adverse environmental conditions

What does aerobic respiration result in?

results in formation of hydrogen ions, converted to toxic hydrogen peroxides as part of the E.T.C.

light repair or photo reactivation

reverses structural changes with exposure to visible light.

What shape is Bacillus/bacilli?

rod shaped

What shape does the lymphocyte nucleus have?

round and takes up most of the cell

the x4 ocular lens is also called?

scanning objective

streptococcus pyogenes

scarlet fever

Mature neutrophils...AKA?

segs

Is PEA selective or differential?

selective

What is the purpose of PEA testing?

selects for Gram + organisms, and is used to isolate them from cultures where they are mixed with Gram - bacteria

Bile Esculin test (BEA)

selects for bacteria that can tolerate bile salts. differentiates species that rely on hydrolysis of exculin to form a dark brown or black color change. positive= growth of bacteria with a color change of the media to black. negative= growth of bacteria without color change or no growth of bacteria.

Eosin Methylene Blue Agar (EMB)

selects for gram-negative rods. differentiates between lactose fermenting and non-lactose fermenting. lactose fermenters form colonies with dark centers and clear borders while the non lactose fermenters form completely colorless colonies. positive= growth of bacteria with color change of colonies that have dark centers (magenta or purple), E coli will have a metallic green sheen. negative= growth of bacteria without color change or not growth of bacteria.

What is the purpose of mannitol salt agar?

selects for organisms that can grow in a 7.5% NaCL enviromment.

What is the difference between semisolid agar deeps, petri plates, and slants?

semisolid agar deeps contain .5% -. 7% agar instead of the 1.5% in Petri plates and slants

A relatively large zone inhibition surrounding a anitmircrobil disk would be interpreted as a what a reaction

sensitive

WALK AWAY PANEL antibiotics are clear this means?

sensitive - good choice

why is important to distinguish streptococcus pneumonia

serious pneumonia the other won't cause ...

SBA

sheep blood agar

citrate utilization

simmons citrate agar tests ability to utilize citrate as a sole carbon source and for presence of alkaline products as a result of citrate utilization. indicator bromothymol blue will detect a pH change of greater than 7.5. positive= green to blue. negative= no color change

this is e aerogenes, this is a positive result for simmons citrate, it is blue

simmons citrate test

this is e coli, it has a negative citrate result. it is green

simmons citrate test

What is a pure culture?

single species or kinds

What is the difference between slant tubes and plates?

slants provide a solid growth surface in a tube form, which takes less room and is easier to transport than plates

Entamoeba histolytica

sleeping sickness

what causes trypanosome

sleeping sickness

If positive test after 72 hours with Snyder test?

slight susceptibility to dental caries

describe a plasmid and some of its characteristics

small circular DNA structure separator from chromosomal DNA bacteria

plasmids

small extra-chromosomal elements that can exist in some bacteria. carry genes for antibiotic resistance, and carry genes for other proteins that may be beneficial

What do the small pores in the gram positive peptidoglycan layer do?

small pores enable them to retain primary dye (crystal violet a purple dye) when samples are washed with alcohol

Why are petri plate cultures inverted when they are in the incubator?

so that condensation doesn't drip on cultures

Station 1 Gelatin test Negative result:

solid

What is the purpose for agar in lab?

solidifying agent

Amylose:

soluble starch

hypertonic

solute concentration is greater outside of the cell. water flows out

hypotonic

solute concentration is lower outside of the cell. water flows in

Results of LB/AMP plate with +pGLO?

some growth of bacteria as it will have amp resistance

MSA - differential

some species also ferment mannitol into acid. this acid production turns pH indicator from red to yellow (mannitol break down)

What organism and legume did we use for nitrogen fixation

soybeans, B. japonicum (Bradyrhizobiuim japonicum)

E. coli JM 83

specially designed for transforming. contains no plasmids, and makes no restriction enzymes.

Describe a simple stain

stains all the bacteria the same way, so you can more clearily see each cell or its specific cell components

Negative stain

stains the background

CATALACE what organism is double zone hemolosis (catalace positive)?

staph aureus

What is the CAMP factor?

streptococcus agalactiae produces hemolysin with staph. aureus.

virulence factors

structures or substances produced by a microorganism which enhance its ability to cause disease

Spores

structures that can survive for long periods of time referred to as "resting bodies" due to their metabolically inert state.

epidemiology

study of patterns of disease infection and spread

How are enzymes named?

substrate name + suffix of -ase Fructose=Fructase Starch=Amylase Glucose=Glucase

What is the critical step in performing the acid fast stain?

successfully forcing the primary stain inside the cells by using heat

In lab 5 (identifying an unknown), which mediums needed to be innolculated with a loop?

sugar fermentation tubes, urea broth tube and nutrient agar slant.

What is the biochemical reaction for TSI?

sugar------------------------> acid with/without gas

What is the purpose of the SIMs test?

sulfur reduction, indole pretection, and motility

How is ferrous sulfide formed?

sulfur-containing amino acids are broken down into hydrogen sulfide which is then combined with iron sulfate in the tube and forms ferrous sulfide.

What is growth medium?

supports growth of bacteria

WALK AWAY PANEL the bottom part is for the?

susceptability

What is reported out to Physicians from the Kirby-Bauer?

susceptibility or resistance

What technique is used for the Kirby-Bauer test?

swab and make a bacterial lawn

What technique was used for UV test?

swab and make a bacterial lawn

what kind of relationship do the organism and the legume have

symbiotic

gresiofulvin, anti-fungal agent

synthesized by some fungi, exact mechanism is unknown, thought to interfere with spindle fiber components (tubulin) and blocks mitotic events. used in treatment of the nail and skin fungal infections (Hint: difficult name - don't know how it works)

What does an increase in neutrophils indicate?

systemic bacterial infection

flucytosine, anti-fungal agent

taken by yeast cells and converted into active inhibitory form, inhibits nucleic acid synthesis (Hint: fluc~fuc~totally screwed~you are totally screwed if you can't make nucleic acid)

What are the primary differences between tapeworms and flukes?

tapeworms possess a head end called a scolex and are segmented into units called proglottids

standard plate count

technique often used to determine the number of bacteria in a sample

Indirect ELISA - what are we testing for?

test for antibodies "Host response"

Phenol Red Broth - purpose?

test for fermentation of carbohydrates and gas production

MSA - purpose of mannitol?

test for fermentation of sugars

Snyder Test - purpose

test for growth of oral bacteria

Catalase Test - purpose?

test for presence of catalase

starch hydrolysis

tests for amylase combined with water breaks down large polysaccharides into smaller molecules that can be used for energy. positive= a clear halo around the colonies after the addition of gram's iodine, negative= the entire medium is dark blue/brown

nitrate reduction test

tests for the ability to reduce nitrate to nitrite. if the organism can reduce the nitrate it will react with the reagent (paba) positive= yellow, pink or red color change after the addition of papa. negative= no color change

urea hydrolysis

tests for urease enzyme that breaks down urea to form two molecules of ammonia. ammonia products form a basic environment that causes the media to change from a light orange to magenta. positive= bright pink slant. negative= light orange to yellow

define resolving power

the ability of a lens system to distinguish two neighboring points as seperate and distinct

What is the purpose of the DNAse test?

the ability of a organism to utilize nucleotides from DNA or RNA as a nutrient source

What does Methyl Red test determine?

the ability of an organism to produce acid end product from glucose fermentation

What does Voges-Proskauer test determine?

the ability of some organisms to produce a neutral end product during glucose fermentation

What does resolution mean?

the ability to distinguish between two adjacent objects

Why should the rheostat never be turned higher than 7?

the bulb burns out too quickly

The size of the zone of inhibition does not indicate ___________

the degree of effectiveness of a chemical agent

CO₂ uptake is related to the __________

the increased growth of anaerobic organisms

Oxygen uptake is directly proportional to __________

the increasing number of vigorously growing aerobic cells

What is the Kovac's reagent used for?

the indole production in SIMs

MONOCYTE

the largest in sizeof WBC 3-7% 3rd place turn into macrophages kidney shaped

What is thermal death time (TDT)?

the length of time required to kill all bacteria at a given temperature

why is it wrong to turn the fine adjustment more than slightly in either direction?

the object could become out of range and it could damage the lenses

What does OF GLUCOSE test determine?

the organism's ability either to oxidize glucose or to ferment glucose

What does Catalase Test determine?

the presence of the enzyme, catalase

what does the oxidase test detect?

the presence or absence of cytochrome C

How is sugar fermentation detected?

the production of acids

what is hemolysis

the rupture or description of red blood cells

What is the critical step in doing a gram stain?

the timing of the decolorizer

presumptive test (acid and gas production)

the tubes that test positive in this test go on to be confirmed using MUG (methylumbelliferyl-beta-D glucoronide) agar or EMB agar

Why do ordinary stains like gram stain not work on acid fast bacteria?

these cells have a waxy lipid cell wall

How would immunosuppressive drugs predispose a patient to opportunistic fungi?

these drugs greatly reduce the natural immune state especially the cellular immunity which is essential in controlling these infections

Why are arthropods important to microbiology?

they represent important agents (vectors) of disease transmission

Describe the gram negative peptidoglycan layer.

thin layer with larger pores, therefore they cannot retain the primary dye and become colorless after being washed with alcohol

kirby bauer test

this depends on the sensitivity of a bacteria to an antimicrobial= zone of inhibition name of inoculation technique? bacterial lawn

Results of LB/AMP with no pGlO?

this is our negative control as there is no growth because there is no plasmid to give it resistance to the AMP

Results of LB with no pGLO?

this is our positive control as there is growth because there is no amp present to stop growth

the one on the left is positive, the one in the center is negative, and the one on the right is an un-inoculated tube

this is simmons citrate test

methyl red test, e coli (mr positive) on the left and e aerogens (mr negative) on the right. page 82

this is the methyl red test.

e. aerogenes is vp positive, this is the red/wine color indicative of a vp positive test.

this is the vp test

e. coli (vp-negative) is on the left and e. aerogenes (vp-positive) is on the right. atlas page 98

this is the vp test

e. coli is vp negative,

this is the vp test

biological transmission

through bite of an infected mosquito, tick, or flea

Why do we place the tubes on ice after the heat shock?

to close the pores of the E. coli so the DNA will stay inside

What is the purpose of the Starch test?

to detect the presence of amylase

What is the purpose of the Simmon's Citrate Agar?

to detect the presence of an exoenzyme known as citrate permease

What is the purpose of the Casein Hydrolysis test?

to detect the presence of the casease enzymes

What is the purpose for Oxidase testing?

to detect the presence of the endoenzyme, cytochrome oxidase

What is the purpose of the nitrate reduction?

to determine if enzyme nitrate reductase was produced to reduce nitrate to nitrite

What is the purpose of the Oxidase test?

to determine if the bacterium of interest has cytochrome C oxidase in its electron transport chain

What is the purpose of the Urease test?

to determine rapid or slow urease

Gelatin Liquéfaction test "Nutrient Gelatin" - Purpose?

to determine the ability of an organisms to produce the enzyme gelatinase

What is the purpose of 6.5% NaCL Broth test?

to differentiate between strep that can grow in 6.5 % salt environment and those that cannot

What is the purpose of the catalase test?

to distinguish between the gram-positive cocci

Why must 9 ml sterile water blanks be used for each dilution rather than 10 ml blanks?

to each sterile water blank, we add 1 ml of specimen, thus the volume is then 10 mls for a 1:10 dilution

Why are bacteria inoculated into various growth media as pure cultures?

to identify bacteria, without introducing unwanted microbes (contaminants)

Decarboxylation Test - purpose?

to identify organims that have decarboxylases to break down amino acids

Why is there an indent in the tube for testing for citrate?

to inhibit 02 from reaching the bottom of the tube

What does subculture mean?

to inoculate a new sterile place from the old culture

Why are selective and differential media used?

to isolate certain bacteria and differentiate one from another by the biochemical reactions they perform

When opening a culture tube, what is the purpose of flaming the mouth of the tube?

to kill any contaminants and to release a puff of air

What are the reasons for heat-fixing the smear?

to kill the bacteria and it also helps the bacteria adhere to the slide

Why are we using GFP?

to show visual evidence that the DNA transformed with successful gene expression

Whats the purpose of flaming the inoculating loop?

to sterilize for aseptic technique

calculate total coliform count

total coliforms/100mL = coliform colonies counted divided by he number of mLs sampled. Multiply the product by 100. Answer in CFU/mL

Define sterile

totally free of organisms

What will negate the charge of DNA in the transformation lab?

transforming solution

citrase

transport citrate into the cell and make it available for energy catabolizing processes.

Describe two methods used to make e.coli competent to up take the plasmid

treated with calcium chloride and used to

would person living in the desert or tropics have a larger number of bacteria living on their skin

tropics

blood agar contain red blood cells

true

TSA

tryptic soy agar plate

Hydrogen sulfide production: Negative result, Positive result, media used & reageant added

tube is less than 50% black, tube is more than 50% black (ferrous sulfide forms), semi-solid media, iron sulfate

Mycobacterium tuberculosis

tuberculosis

Oxidase Test - positive test?

turn bacteria violet to purple immediately

what role does arabinose play in transformation

turns on the green/ inducerin recombanted DNA

what is a mixed culture?

two or more kinds

Compound Microscope

two or more lens to view an object

Selective & Differential

type of media Mannitol/Salt agar is

Replica plating

type of plating used to isolate bacterial mutants

What does the medium contain for the Urease test?

urea phosphate buffer and phenol red

fine adjustment knob

used for 40x and 100x (oil immersion) lenses

indicator organism

used to determine if there is fecal contamination. this indicator is present in human waste in high enough numbers to be readily detected and easy to find.

MR test

used to identify bacteria that produce stable acid end products by means of mixed acid fermentation of glucose. a pH indicator methyl red is added and in acidic environments will turn red. neutral environments it will be yellow. positive= red, negative= no color change

Fine adjustment knob?

used to sharpen the focus, don't turn too many times objectives may crash into the slide

Citrate Utilization - what is going on?

uses citrate breaks down the ammonium salts to ammonia, which creates an alkaline pH

Differential stain

uses more than one dye and stains different kinds of organisms different colors

Citrate Utilization (SC) - purpose?

using citrate as the sole source of carbon with production of enzyme citratase

if a microbe can produce cats

water oxygen

normal flora

we harbor normal flora in and on us some of these microbes are potentially pathogenic (disease causing)

understand the regulation of the pGLO plasmid

we make the cells competent through addition of CaCl2 and heat shock, making them able to take up naked DNA (plasmid). those that take up the plasmid will be able to grow on a TSA plate with ampicillin present.

TSI- Triple Sugar Iron

what are the three sugars used?- glucose, lactose, and sucrose yellow means acid fermentation yellow with bubbles or tears in agar= acid fermentation and gas end products black=sulfur reduction red= alkaline yellow color change= E.coli Red color= Alcagenes

Negative

what kind of staining is this?

Phenol red broth

whats the name of the tube used? Durham tube what organism caused the yellow tube and gas? E.coli what organism caused no color change? Alcagenes indicator: phenol red

When does endospore reproduction occur?

when bacteria do binary fission

How are spores produced?

when environmental conditions are poor, meaning essential nutrients and water are not available to the cell.

What are oxidizers?

when glucose is utilized only in the presence of O2

When are bacteria considered to pigmented?

when it has more density it will appear to be black or dark brown

define parfocal

when one lens is in focus, the other lenses will be in focus with only minor adjustments to bring the object into sharp focus

the one with the pink is positive for the indole test. it has tryptophanase which will break down the tryptophan amino acid into indole/ammonia/pyruvate. the Kovac's reagent (DMB(reacts with free indole to form complex under acidic conditions)/amyl alcohol(allows complex to float on medium)/HCl(provides acidic conditions) is used to detect this

which one is positive for the indole test and why?

Voges-Proskauer "MR-VP Broth" - positive result?

wine red color develops

aseptically

without introducing other microbes

What are the 4 ocular lens settings?

x4, x10, x40 and x100

If the optic lens is x10 and the ocular lens is set at x40 what is the total magnification?

x400

Phenol Red Broth - positive test?

yellow

Methyl Red Test "MR-VP Broth" - negative test?

yellow color develops

Phenylalanine Deamination "Phenylaline Agar"- negative test?

yellow color develops

Mannitol Salt Broth positive and negative

yellow is positive - presumptive S. aureus red is negative - presumptive non-pathogenic Staphylococcus

Tryptophan Hydrolysis "Indole Test" - negative test?

yellow layer forms on surface of medium

Station 14 TSIA Third tube

yellow slant/yellow butt and agar lifted - glucose and Lactose and/or Sucrose fermentation and acid accumulation in slant and butt, also produced gas.

What observations are made in lab with Kirby-Bauer?

zone of inhibition

Basophils

~Appear in many specific kind of inflammatory reactions, particularly those that cause allergic symptoms

Platelets

~Blood clots ~Store and transfer chemicals

Monocytes

~Can easily be transported to the site of infection (Host defense)

Red Blood cells

~Carry Oxygen from lungs to tissues and other organs ~Getting waste carbon dioxide from your tissues to your lungs

Neutrophils

~Inflammatory events ~Phagocytosis (ingest and kill micro-organisms)

Eosinopils

~Movement to inflamed areas ~Trapping substances ~Killing cells ~Antiparasitic and bactericidal activity ~Participating in immediate allergic reactions ~modulating inflammatory responses

Lymphocytes (T cells and B cells)

~T cells can only kill unhealthy cells and cannot kill pathogens that live outside the cells ~B cells produce antibodies

On a blood agar plate, what does a green zone around colonies indicate?

α-hemolysis

On a blood agar plate, what does a clear zone around colonies indicate?

β-hemolysis

On a blood agar plate, what does no zone around colonies indicate?

γ-hemolysis

Pseudomonas aeruginosa

• motile (one or more polar flagella) • rod shaped and aerobe • Gram-NEGATIVE bacteria. They are found almost everywhere, in soil, water, plants and animals. • Opportunistic POSITIVE CATALASE RESULTS = Liberation of Bubbles

salmonella enterica: General Characteristics

•Enteric pathogen •motile •lactose non-fermenter •H2S producer •Ferment glucose by one of two major pathways to a variety of end products

Mannitol Salt Agar (MSA)

•selective/differential •selects (salt) inhibits most G+/G- bacteria from growing •differentail (mannitol) mannitol can fermented by S. aureus, so this media differentiates S. aureus from other staph (becuase it is the only on that can ferment mannitol) •mannitol fermentation results in acid production, pink → yellow •when you streak for isolation in this media, a section of it will remain pink, why? becuase the isolated colonies don't have enough bacteria to produce enough acid to perform a color change like on the rest of the plate, or colonies simply did not grow there at all

Methyl Red Test "MR-VP Broth" - products from the breakdown of formic acid?

CO2 and H2

What is a faculative anaerobe?

Can grow with or without oxygen, but grow better in oxygen so they will grow through out the tube but mainly on top.

Starch Hydrolysis

Can this bacterium make the enzyme amylase?

Yeast that causes thrush

Candidia albacase

MSA- Mannitol Salt Agar

Carbon source= Mannitol indicator: phenol red positive organism= S. aureus yellow halo means= mannitol fermenter

Station 13 Casease test Enzyme

Casease

Station 13 Casease test Know the protein in milk (substrate)

Casein

CATALASE TEST

Catalase is an enzyme that converts hydrogen peroxide into water and oxygen. The presence of catalase can be detected by the slide method. A drop of 3 percent hydrogen peroxide is put on a slide and the bacteria is emulsified in the drop. The presence of bubbles is evidence of the production of OXYGEN. Alternatively, bacteria may be tested by dropping hydrogen peroxide directly onto colonies growing on a plate.

Why do we add zinc dust to a nitrate test if no color change occurs after the reagents?

Catalyzes the reduction of any nitrate to nitrite, producing a red color

Mixed acids

Causes Methyl Red to turn red

Station 10 Parasitic worms Class

Cestodes, Tapeworms, leaches ect.

Mutation

Changes in the genetic material

Hemolysin

Chemical that can break down red blood cells

Fungi characteristics

Chitin in cell walls, unicellular or multicellular, lack chlorophyll, heterotrophs

What's the purpose for CAMP test?

Christie, Atkins, Munch-Peterson is used to differentiate Streptococcus agalactiae.

Organisms lacking _____ will not survive on the Simmons citrate medium.

Citrate-permease

-Aspergillus -Candida -Coccidioides -Histoplasma ^^^^all are ASCOMYCETES and appear to form ascospores as a sexual means of reproduction -Deuteromycetes = all other fungi that sexual means of reproduction is unknown

Classification of Mycoses

Match the disease causing organism listed on the left with the description or disease on the right. Answers may be used more than once. Claviceps purpurea a. Malaria Toxoplasma gondii b. Sheep liver disease Plasmodium falciparum c. Ergot poisoning Fasciola hepatica d. Hookworm Necator americanus e. Cat feces disease

Claviceps purpurea = C Toxoplasma gondii = E Plasmodium falciparum = A Fasciola hepatica = B Necator americanus = D

Station 13 Casease test Positive result:

Clearing around the growth, means the bacteria produced casease, so the protein in milk casein will be digested

Station 12 Starch test Positive result:

Clearing around the organism, means amylase was produce and starch was broken down.

Bound coagulase is also called:

Clumping factor

***If plasma becomes clumpy and or solidifies, then bacteria are coagulase positive **Test is only valid on gram + staphylococcus like bacteria since gram negative bacteria are able to provide false positive reactions from a non coagulase like mechanism. ****Staphylococcus aureus has clotting virlence factor, causeing the solidification on the media.

Coagulase test

The Novobiocin test is used to differentiate:

Coagulase-negative staphylococci

Bacillus subtilis -Colonies of B. subtilis grown on a culture dish in a molecular biology laboratory.

Colonies of B. subtilis grown on a culture dish in a molecular biology laboratory.

Red

Color Voges-Proskaeur test turns when positive

Blue

Color seen in citrate test when pH is higher than 7.6

Green

Color seen in citrate test when pH up to 7.5

Pink

Color urease agar turns when urease is present

Red= negative sugar fermentation, negative H2S, Yellow= positive sugar fermentation Black= presence of hydrogen sulfide

Colors and what they signify in the result of a triple sugar iron agar test

Red = negative, Yellow = positive

Colors that signify positive or negative in a Phenol Red carbohydrate fermentation test

Compare the geographic distribution of: -Candidiasis= entire United States -Filobasidielliosis (Cryptococcosis)= entire United States -Histoplasmosis= Mississippi River Valley regions -Coccidioidomycosis= Western States; Sonoran Valley; West TX

Compare the geographic distribution of: -Candidiasis -Filobasidielliosis (Cryptococcosis) -Histoplasmosis -Coccidioidomycosis

Peptone, glucose, Phosphate buffer

Contents of MRVP broth

Hydrogen Sulfide Production "Triple Sugar Iron Agar" Enzyme

Cysteine Desulfurase

Enzyme that is an electron carrier in the electron transport chain. Present in bacteria that utilize oxygen.

Cytochrome oxidase

1 ml of a 1:5 X 10^5 dilution of milk was plated in duplicate. Plate A contained 230 colonies, plate B contained 270 colonies following incubation. How many bacteria were present in a milliliter of the original sample?

DF = 500,000 TVB/ml = PC X DF PC = 230 = 250 X 500,000 270 = 125,000,000 500/2 = 250 Ave PC TVB/ml = 1.25 X 10^8

What medium is used for DNAse?

DNAse plate: nutrient agar with DNAse added

**Tests for exoenzyme DNase which is able to hydrolyze DNA ***Zones of clear around streaks either before or after addition of 0.1N HCl is a positive result for the presence of DNase

DNase test

What is the most important step in a differential stain?

Decolorization (alcohol) because at this point you can distinguish between 2 bacteria. For example: Gram + have a violet color after this step and Gram - are colorless.

***Selects for gram -, lactose - microorganisms **Some Lactose + colonies do grow but they will appear Red

Desoxycholate Citrate test

Indole Production Test

Determines the ability of microorganisms to hydrolyze tryptophan Kovac's Reagent -- red reagent layer = Positive

The purpose of a standard qualitative analysis of water

Determines the presence of coliform which indicates fecal contamination

Is Phenol Red Broth selective, differential, or enriched?

Differential for Enterobacteriaceae

Is Simmons citrate selective, differential, or enriched?

Differential for facultative anaerobes

Importance of using blood agar

Differentiates ability to lysis the red blood cells and provides nutrient

Corynebacterium Diphtheriae

Diphtheria

No

Does micrococcus luteus cause hemolysis?

Yes

Does staphylococcus aureus cause hemolysis?

oxidase: Negative result (before 20 sec), Positive result (before 20 sec), media used

Dryslide remains colorless, dryslide turns blue/purple (cytochrome C is present), nutrient agar slant + oxidase dryslide

Which gram negative organism would result in a metallic green sheen after culturing in a EMB agar

E. coli

Least resistant bacterium:

E. coli and S. aureus

Why is E. coli more susceptible to boiling than B. pumilus?

E. coli lacks the endospores that B. pumilus possesses

What organism produces pink colonies when grown on Maconkey agar? found in a water test?

E. coli; fecal decontamination

What media is used to grow Gram negative enteric bacteria?

EMB, Hektoen Does not grow gram + due to bile salts and some dyes.

Which selective medium would be best to determine the amount of acid produced by gram negative bacteria

ESA methylan blue agar

Hemophilus Influenza

Ear infections, bronchitis, pneumonia, meningitis

This tapeworm can form large hydatid cysts virtually anywhere including the lungs and brain.

Echinococcus granulosis

These organisms are typically oxidase negative:

Enterobacteriaceae

These eggs can hatch into large intestinal worms that will feed on the victim's blood like Dracula.

Enterobius vermicularis

ELISA

Enzyme Linked ImmunoSorbant Assay

Cytochrome Oxidase

Enzyme present if the paper in a Cytochrome oxidase test turns purple/blue

Gram +

Eosin Methylene Blue (EMB) agar inhibits growth of this type of bacteria

EMB

Eosin Methylene Blue Agar

Selective and differential Contains eosin and methylene blue dyes to inhibit gram (+) bacteria Contain lactose and sucrose to distinguish bacteria that ferment and don't ferment

Eosin Methylene Blue Agar

What is name for a condition with a lot of Eosinophils in the blood?

Eosinophilia

What kind of white blood cells are triggered by allergic reactions and parasitic infections?

Eosinophils (phagocytic)

Widespread occurence of an infectious disease in a community at a particular time

Epidemic

2H2O2 > 2H2O + O2 (Hydrogen Peroxide > Water and Oxygen)

Equation for the reaction of catalase

Which bacteria has mechanisms to repair damage from UV radiation?

Escherichia coli

What happens to produce a positive result in a Bile Esculin Test?

Esculin molecules split, esculetin reacts with Fe3+ from ferric citrate and forms a dark brown precipitate

Pseudomonas aeruginosa

Example bacteria that is Cytochrome oxidase positive

Hemolysin

Exotoxin that lyses RBC

True/False: All of the protists exhibit an ability to form antibiotic resistant structures called cysts.

False

True/False: All protozoans have complex methods of reproduction such as seen with malaria.

False

True/False: Animal parasites such as roundworms reproduce most commonly by producing asexual spores.

False

True/False: Antibiotic sensitivity plates are covered with bacteria using the loop and following the dilution streak plate technique.

False

True/False: Ascaris lumbricoides is the smallest roundworm and measures only 1mm in length.

False

True/False: Biological vectors would include ants and houseflies.

False

True/False: Body lice only transmit diseases mechanically and are therefore harmless vectors.

False

True/False: EMB is the most common culture media used in the Kirby Bauer sensitivity test.

False

True/False: Fortunately animal parasite worm infections are not very common in the world today.

False

True/False: Fungal infections are commonly spread through the air by means of their cysts.

False

True/False: If an organism is sensitive to one antibiotic it will automatically be sensitive to others.

False

True/False: Infections caused by fungi are commonly known as helminth infections.

False

True/False: Malaria is caused by species of Leishmania.

False

True/False: Ringworm can be treated with anti-worm medication such as mebendazole.

False

True/False: Ticks absorb blood directly from the victim's intestinal tract.

False

True/False: To ensure elimination of tapeworm infections, the proglottids must be removed from the host.

False

True/False: Trichinosis is a common pinworm infection affecting children most of the time.

False

True/False: Zone diameters of sensitivity are measured in inches to be as accurate as possible.

False

This worm is acquired from contaminated deep fried chicken livers.

Fasciola hepatica

What does EMB isolate?

Fecal coliforms

Coliforms

Fecal contamination is identified by finding ______ in the water.

Trypanosoma gambiense

Flying Birds

a. they produce resistant cysts. b. they are antibiotic resistant c. *they produce infectious spores* d. they take steroids and are tougher than we are

Fungi are dangerous to work with in the lab because: a. they produce resistant cysts. b. they are antibiotic resistant c. they produce infectious spores d. they take steroids and are tougher than we are

Proteus vulgaris

GNR

Pseudomonas aeruginosa

GNR

Salmonella

GNR

Serratia marcescens

GNR

Shigella

GNR

Corynebacterium

GP pleomorphic rod

Enterococcus

GPC

Streptococcus

GPC

Staphylococcus aureus

GPC "in clusters"

Strep pyogenes

GPC in chains

Strep agalactiae

GPC in long chains

Streptococcus pneumonia

GPC in pairs

Staph epidermidis

GPC in pairs and tetrads

Staph saprophyticus

GPC in single or pairs

Bacillus megaterium

GPR

Clostridium

GPR

In a TSIA slant, cracks or lifted medium indicate:

Gas production

Cracks, Lifting

Gas production can be determined by ------- and/or the -------- of the media off the bottom of the tube.

This is the only protozoan that has functional eyes which it uses to attack and bite its next victim.

Giardia intestinalis and G. lamblia

What provides the carbohydrate in MR/VP broth?

Glucose

In a TSIA slant, a yellow slant/yellow butt indicates:

Glucose fermenter, and fermenter of one of the other sugars

In a TSIA slant, a red slant/yellow butt indicates:

Glucose fermenter, no lactose or sucrose fermentation

Obtain an entire yard of bacteria

Goal of a spread plate

Obtain individual colonies of bacteria

Goal of a streak plate

Neisseria Gonorrhoea

Gonorrhea

Properties of a COLIFORM

Gram (-) Non spore forming Bacillus - rod Lactose + and produces gas/acid

Negative

Gram ___________ intestinal pathogens are a major concern for public health since the two main pathogens Salmonella and Shigella have the ability to cause enteric fevers, food poisoning, dysentery, and even typhoid fever

What color is each kind of bacteria after ethanol is added (and washed off)

Gram+ = purple Gram- = colorless

Is Catalase used for Gram-positive or Gram-negative organisms?

Gram-positive

What are the two types of white blood cells?

Granulocytes & Agranulocytes

Do bacteria have a greater or smaller variety of metabolic processes than eukaryotic organisms?

Greater

Station 2 Phenol Red media Cap colors:

Green - Lactose Light pink - sucrose Red - glucose

Station 9 Citrate test Postive result:

Growth on slant, turns blue

What is the biochemical reaction for Ferric Ions?

H2S + ferric ions-----------> ferrous sulfide

Who formulated the Gram Stain?

Hans Christian Gram (1884)

Scolex

Head of a worm

What is the mordant in acid-fast stains and endospores?

Heat because heat drives the stain into the cell/cell wall.

What is the main difference between an acid fast stain and a Gram stain?

Heat must be applied to an acid fast stain to hold in the stain

Clear around colonies

How amylase positive bacteria appear on Amylase positive test result

-Oral candidiasis = Nystatin or Amphotericin B -Taenia pedis = Griseofulvin -Filobasidielliosis (Cryptococcosis) = Amphotericin B -Ringworm = Griseofulvin, Mycosyl -Aspergillosis = Amphotericin B -Pneumocystis pneumonia = Trimethoprim/Sulfamethoxazole and/or Pentamidine

How are the following mycoses treated? -Oral candidiasis -Taneia pedis -Filobasidielliosis (Cryptococcosis) -Ringworm -Aspergillosis -Pneumocystis pneumonia

Binary fission

How bacteria reproduce

Abstinence or condom use Not sharing needles Using gloves and masks (medical staff)

How can HIV be avoided?

-these pathogenic yeasts have been known to colonize birds -the yeasts can survive for long periods of time in bird droppings

How could pigeons or starlings contribute to the spread of Filobasidiella neoformans?

Involves enzymes detected by endonucleases. DNA polymerase I fixes it & ligase closes the gap.

How does Dark-repair (excision repair) fix DNA?

Small amount

How much blue color is necessary to indicate that a citrate test is positive?

Clonorchis sinensis

Human Liver Fluke

pH indicators and respective medium and color produce?

Hydrogen Sulfide Production "Triple Sugar Iron Agar" Phenol Red

WALK AWAY PANEL the top part is for the?

ID

Glucose + Lactose + H2S - Escherichia Enterobacter

Identify this KIA tube

Glucose + Lactose - H2S + Salmonella Proteus

Identify this KIA tube

Glucose + Lactose - H2S - Shigella ****NO gas in KIA butt

Identify this KIA tube

Glucose - Lactose - H2S - Pseudomonas Alkalingenes

Identify this KIA tube

Facultative

Identify this OF Glucose tube

No respiration

Identify this OF Glucose tube

Strictly Oxidative

Identify this OF Glucose tube

Strictly fermentive

Identify this OF Glucose tube

What are TSI slants used for?

Identifying Gram (-) bacilli

Cysteine desulfurase

If a bacteria contains the enzyme ------------, a black precipitant will form

Station 12 Starch test Principle

If bacteria produces amylase, it breaks down the substrate starch.

When do we test for antibodies?

If someone was exposed a long time ago

When do we test for antigens?

If someone were to get stuck with an infected needle

What is the purpose of Methyl Red and Voges-Prokauer (MRVP) ?

If the end products of carbohydrate metabolism is acidic or neutral. MR - Mixed Acid Fermentation VP - Acetone

How soon can the MR test be read after adding reagent?

Immediately

when did the practice of washing hands become significant?

In 1846 the method became significant as a method of infection prevention

Negative, Green, Blue

In OF Glucose test, no color change results in a _______ result. Brom Thymol Blue is actually _______, but turns _______ in highly negative results (high basic pH)

Brom Thymol Blue

In OF Glucose test; media contains pH indicator ---------, which turns yellow if the glucose is utilized, and acids are produced.

Mineral oil

In OF Glucose test; two tubes are inoculated, with one of the tubes covered in -------- to prevent air from reaching the media.

Bubble

In the catalse test, bacteria that are + cause hydrogen peroxide to do this

Phenol Red

Indicator that is sensitive to pH and turns yellow in the presence of acid.

Litmus milk test

Indicator: Litmus pink= lactose fermentation what organism caused pink color? e.coli white= litmus reduction what organism caused white color? enterococcus blue= hydrolyze casein basic

In a SIM test, no red color after Kovac's reagent indicates:

Indole negative

In a SIM test, a red color after Kovac's reagent indicates:

Indole positive

Beta-Lactams

Inhibit cell wall synthesis Ex: Penicillin G

Sulfamides

Inhibit synthesis of nucleic acids Ex: Sulfamethoxazole

Tetracyclines

Interfere with translation by bonding to ribosomes

What reagent is used for Starch test?

Iodine

Positive

Is patient A positive or negative for Clostridium difficile?

Negative for Streptococcus

Is this test positive or negative? For what?

What is a major limitation to UV radiation?

It cannot pass through solid surfaces

How does the oxygen dryslide change colors?

It contains a chromogenic substrate that oxidase-positive bacteria will change blue

Why can't a disinfectant be ingested?

It contains harmful chemicals that will breakdown our natural protective barriers.

Where does the acid come from in the sugar fermentation test?

It is an end product of the break down of sugar

What is needed to obtain bioluminescense?

It is positive for pGLO and the arabinose activates the GFP causing it to glow.

Station 7 Kirby Bauer test Know the name

Kirby Bauer Test

GIve another name for antibiotic Susceptibility Test?

Kirby-Bauer

What method is used for the antimicrobial susceptibility test?

Kirby-Bauer

Antimicrobial Susceptibility Test..AKA

Kirby-Bauer or disk diffusion test

What organism would be on the capsular stain?

Klebsiella

Tests for ability to ferment glucose and/or lactose, tests H2S production, pretty much the same thing as RDS test but the difference is that KIA contains iron salts to determine H2S production.

Kligler's Iron Agar (KIA) test

What reagent is used to indicate the presence of indole?

Kovac's Reagent

What reagent is used for the SIM?

Kovac's reagent

Station 3 Indole test Reagent

Kovacs

What type of agar is used for Bacterial Transformation lab?

LB - Luria Bertani

MAC agar - purpose of lactose

LF - lactose fermentation NLF - no lactose fermentation

Bacteria are most sensitive to chemicals in which stage of development?

LOG stage (they are not actively growing)

What are the bacterial species involved in yogurt production

Lactobacillis bulgaricus for flavor/aroma and continues fermentation, Streptococcus thermophilus for texture and initial fermentation

Hot pink color means what in MacConkey agar?

Lactose Fermentation

What are the 3 kind of sugars in TSI?

Lactose-a large amount Sucrose Glucose-a small amount

Phenylalanine Deaminase

Looking for the presents of Phenylpyruvic acid.

Fucus

Looks like a tree with bulbs on the end!

Ulva

Looks like boiled cabbage!

Entamoeba histolyctia

Looks like tissue!

Name at least 2-3 things that need to do transformation experiment?

Lurea Bertani Medium Agar; Transforamtion solution; heat shock and Ice

Extra credit

Lyme diseases-----Borrelia burgdosferi Anthrax----- Bacillus anthracis Tetanus---- Clostridium tetani Cholera---- Vibrio cholerae Tuberculosis----Mycobacterium tuberculosis Gonorrhea---- Neisseria gonorrheae Syphilis ----Treponema pallidum Strep Throat----Streptococcus pyogenes.

What kind of white blood cells are for the immune system?

Lymphocytes

***Enriched media containing 10% NaCl, which selects for Staphylococcus since Staphylococcus prefer the higher salt concentration, which inhibits most other organisms

M-staphylococcus broth test

What does a red medium indicate in Methyl Red testing?

MR +

What does a yellow to orange medium indicate in Methyl Red testing?

MR -

What is the medium used in Methyl Red?

MR/VP

•Both selective and differential •contains 3 ingredients: Lactose (D); Bile (S); Crystal Violet (S) •Lactose + colonies are PINK •Lactose - colonies are COLORLESS •G-GI pathogens have NO problems growing here

Mac CONKEY AGAR:

clockwise from top: e. coli, e. aerogenes, shigella sonnei, proteus mirabilis. e. coli and e. earogenes produce pink color from acid producing lactose fermentation. p. mirabilis, lactose non-fermenter remains normal color

MacConkey agar

Purpose of the Revolving Nosepiece on Brightfield Microscope

Makes the microscope "parfocal"; allows each of the 4 objective lenses to be turned into position for viewing.

Station 16 Malaria Name of the disease

Malaria

falciparum

Malaria

Purpose of the Mechanical Stage Control Knobs on Brightfield Microscope

Maneuvers slide around to change viewing.

Phenol Red Fermentation

Media : 0.5 % Glucose solution or 0.5% Mannitol broth with indicator phenol red . Procedure : broth is inoculated with test bacteria and incubated at 98F /37C For 24 hours and color noted Theory : Phenol red broth is a test medium to which a carbohydrate like glucose or mannitol is added . Phenol red is red under nuetral pH but tends to turn yellow under acidic condition. Acid production from fermentation of the sugar lowers the pH and medium turns yellow indicating carbohydrate utilization by the bacteria . (A) Gas production also from fermentation can be detected as a bubble in an Inverted Durham's tube inside the broth. (AG)

INDOLE RING TEST media

Media : 1% tryptone broth is used . It contains amino acid tryptophan .

CATALASE TEST MEDIA

Media : Bacteria for this test can be grown on nutrient agar TSA PLATE OR** SLANT and the growth is transferred onto a slide to perform this test.

OXIDASE TEST

Media : Bacteria is grown on Nutrient agar and colonies are picked up with a cotton swab and tested with oxidase reagent . Procedure : Using a sterile swab, remove 2-3 colonies from each culture to be tested and smear on a piece of filter paper. Add a drop of the oxidase (TPD) reagent to each spot. If the organism has OXIDASE ACTIVITY, it will turn PURPLE within 30 seconds.

Kirby Bauer Method

Method that allows the rapid determination of the efficacy of a drug -measure zone of inhibition that results from diffusion of the agent into the medium surrounding the disk

Broth/Incubation

Method used to perform carbohydrate fermentation tests

Stab and Streak

Method used to perform urease test

What is the pH indicator of Methyl Red?

Methyl Red

What is the reagent in the MR test?

Methyl Red

MR/VP stands for:

Methyl Red and Voges-Proskauer Broth

Red tube =+. Acids.

Methyl Red test. Which one is positive? For what is it positive?

What is the reagent for testing Methyl Red?

Methyl Red/8-10 drops

Micrococcus luteus

Micrococcus luteus does not grow in the presence of bile salts and crystal violet. •Mannitol Salt Agar (MSA) Test- Positive growth but non-mannitol fermenters includes: Micrococcus luteus • Blood Agar: Gamma Results, no change of mediums color • Biochemical: Catalase Test = POSITIVE A Positive test is indicated by the production of gas bubbles or the presence of effervescence. STAPHYLOCOCCUS and MICROCOCCUS are examples of genus of catalase-positive bacteria.

Gram + Staphylococci

Most bacteria are prohibited from growth on Mannitol/Salt agar except this one

??

Most people find these bugs tasty when dipped in chocolate.

Bifidobacterium, enterocuccus, lactobacillus (anaerobes) E. coli, enterobacter, citrobacter, proteus (facultative anaerobes)

Mutalistic bacteria

Bacteroides, bifidobacterium, enterococcus, lactobacillus, E. coli, enterobacter, citrobacter, proteus

Mutualistic bacteria of the GI tract

Which 2 genre have mycolic acid in their cell walls?

Mycobacteria (causes TB and leprosy) & Nocardia (causes skin infections)

Acid Fast Bacteria

Mycobacterium smegmatis contains mycolic acid

-divided into major groups according to tissue involved in infection -best known to produce mycotoxins or hallucinogenic compounds -some are dimorphic (exhibit as filamentous organisms in nature and as yeast cells in infected tissues) -major spoilers of fruits, vegetables, and grains

Mycoses

-the asexual spores of the mycotic agents are transmitted thru the air and inhaled by victim (however some agents may be acquired via direct contact) -spores can often develop under low pH, high osmotic conditions, and low moisture content which are normally unfavorable for many bacteria

Mycoses Transmission

NEVER LET MONKEYS EAT BANANAS

NEUTROPHILS LYMPHOCYTES MONOCYTES EOSINOPHILS BASOPHILS

Enterobius vermicularis; egg

Name this organism and its form.

Giardia intestinalis; trophozoite phase

Name this organism.

Trypanosoma brucei

Name this organism.

Station 6 Catalase test Name and activity on the enzyme

Name: Catalase Activity: Breakdown its substrate hydrogen peroxides into the water and oxygen gas

These worms "hook" onto your clothing so they can be distributed around the country.

Necator americanus & Ancylostoma duodenale

Mycobacterium

Neg or variable for Gram Rxn-unique cell wal

A green result in a citrate test indicates:

Negative

A solid result for a gelatin test indicates:

Negative

A yellow or orange result for the MR test indicates:

Negative

In a Coagulase Slide test, no clumping indicates:

Negative

In a Coagulase Tube test, a liquid medium indicates:

Negative

No bubbles present in a catalase test indicate:

Negative

No change to the medium in a Bile Esculin Test indicates:

Negative

No color change or copper in the result for VP indicates:

Negative

No color within 20 sec. on an oxidase test is indicative of:

Negative (Cytochrome c oxidase absent)

In a SIM test, no color change of the medium indicates:

Negative (No sulfur reduction)

A yellow result for a urea test indicates:

Negative (urease)

In a SIM test, no radiating growth from the stab line indicates:

Negative motility

Fastidious Bacteria

Neisseria & Haemophilus

Station 11 Parasitic worms Phylum

Nema Toda

Station 11 Parasitic worms Class

Nema Todes

What are the 3 classes of Helminths?

Nematodes Trematodes Cestodes

What is name for a condition with a lot of Neutrophil in the blood?

Neutrophilia

Granulocytes

Neutrophils Eosinophils Basophils

What kind of white blood cells are the most abundant?

Neutrophils (increase during bacterial infection)

INDOLE RING TEST

Next day, Kovac's reagent (p-dimethylaminobenzaldehyde) is added to a broth with indole in it, a dark pink ring develops on top of the tube due to enzyme POSITIVE tryptophanase activity.

What are the reagents in a nitrate test?

Nitrate Reagent A Nitrate Reagent B

Station 9 Citrate test Reagents added after incubation

No

can any chemical completely kill all of the microbes in the area where it is applied?

No

Station 6 Catalase test Negative result:

No bubbles.

Station 13 Casease test Negative result:

No clearing around the growth

Sugar fermentation: Negative result, Positive result, media used & reageant added

No color change and no gas released, gas released and color changes, liquid broth, phenol red

What occurs in γ-hemolysis?

No destruction of RBCs

In a TSIA slant, a red slant/red butt indicates:

No fermentation, utilization of animal proteins, alkaline medium

What would happen if we had a plate that had ampicillin and broth to enhance culture, but the sample did not have pGLO

No grow and no glow

Station 12 Starch test Negative test

No growth around clearing.

Starch hydrolysis: Negative result, Positive result, media used & reageant added

No halo/agar plate remains all blak, clear zone/halo forms around bacteria, starch plate, special iodine

Gelatin hydrolysis: Negative result, Positive result, media used

No liquid formation, liquid formation (gelatinase forms), semi-solid medium

Hydrogen Sulfide Production "Triple Sugar Iron Agar" Negative

No precipitation

Station 3 Indole test Negative result:

No red/ pink ring on top. No color change.

Are endospores a means of reproduction?

No, but are survival dispersal stages of the bacteria allowing them to survive a long time outside of their required living conditions.

To identify the source of gas in a nitrate test, the organism must be a known:

Non-fermenter

MAC agar - LF examples

Normal flora - E.coli and Enterobacter "coliform"

Tests if a bacteria can use glucose in an oxidative (aerobic) or fermentative (anaerobic) condition.

OF Glucose test

Yellow

OF Glucose tube turns ______ if glucose is being used under aerobic or anaerobic conditions

One use of the test is for the preliminary identification of Psedumonas (LACTOSE NEG.) species, which are both oxidase positive and gram-negative.

OXIDASE TEST:Citrate agar is inoculated with Salmonella typhimurium , the medium turns royal blue. This is a positive result for the citrate test.

In a TSIA slant, a red slant/NC butt indicates:

Obligate aerobe, utilization of animal proteins in presence of oxygen, alkaline slant

Red

On SS Agar, all lactose positve colonies appear which color?


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