NWHSU Micro Lab
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| Define titer | The highest dilution of a sample that tests positive for an antibody.
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| Which immunoglobulin appears first when someone has an infection? | IgM
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| Define Prozone. | During antibody precipitation tests, its the period of time when antibody concentrations are high and no precipitates are formed, leading to false negatives.
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| How is a positive Western blot for HIV determined? | There will be two or more dark bands.
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| what are the reagents in gram staining? | Crystal violet, Grams iodine, acetone-alcohol, safranin.
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| In gram staining, what does crystal violet do? | stains all gram + and - bacteria.
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| In gram staining, what does grams iodine do? | its a mordant that forms violet-iodine complex that binds more.
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| In gram staining, what does acetone-alcohol do? | It decolorizes gram - bacteria. But only if not done for too long.
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| In gram staining, what does safranin do? | counterstains gram - bacteria.
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| What color are gram positive bacteria? | Purple
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| What color are gram negative bacteria? | pink
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| How do gram stains and wet mounts vary? | Gram can determine gram +/-. Wet mounts can tell motility.
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| How are gram stains and wet mounts similar? | They both can see arrangement, size, and shape.
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| What is the KOH test to differentiate gram positive from gram negative bacteria? | Mix KOH with a colony for 20 seconds. Stringy=negative. Not stringy=positive.
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| Define ubiquity | Microorganisms everywhere.
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| What are 3 spore forming bacteria that cause disease? | Bacillus Anthracis, Clostridium tetani, Clostridium botulinum.
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| What bacteria causes anthrax? | Bacillus Anthracis
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| WHat bacteria causes Lock jaw | Clostridium tetani
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| What bacteria causes botulism | Clostridium botulinum
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| Define alpha hemolysis and what does it look like? | Bacteria that can partiatlly break down heme pigment (green halo)
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| Define beta hemolysis and what does it look like? | bacteria that can break down heme pigment fully. (clear zone)
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| Define gamma hemolysis and what does it look like? | bacteria that show no change to a blood agar plate.
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| Which hemolysis is a lab looking for in diagnosing strept throat? | beta hemolysis
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| What is the importance of a capsule? | It protects against microphages in lungs.
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| What stain shows a capsule? | gin stain.
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| what is the name of the acid-fast stain and what is it used for? | Ziehl-Neelson procedure uses carbol fushsin. It detects bacteria with cell walls rich in lipids and waxes. LIke Mycobacterium and Nocardia. Leprosy, TB,
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| Define selective media? | Isolates/cultivates for specific bacteria
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| Define differential media? | Shows chemical reactions of bacteria
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| Is MacConkeys selective or Differential? | Selective
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| Is blood agar selective or differential? | Differential
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| What is the catalase test for and what reagent does it use? | H2O2. Catalase breaks H2O2 into H2O and O2. Differentiates streptococci (cat-) from staphlococci (cat+)
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| What is the coagulase test for? | Coagulase enzyme that breaks fibrinogen to fibrin. Staph aureus (coag+). Other staphs (coag-)
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| How is carbohydrate differentiation determined? | Phenol red is used to determine a pH change. Red=no change, Yellow= pH drops and fermentation of carbohydrate has happened. CO2 gas may also be produced
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| HOw does a positive hydrogen sulfide test appear? | Dark precipitate forms with Iron when hydrogen sulfide is present.
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| How does a positive urease test appear? | Pos=bright pink. Neg=salmon->yellow
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| How does positive citrate test appear? | Pos=blue. Neg=green
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| Given a bacterial plate count from a urine culture be able to decide if there is a UTI. | (#colonies)x (_mL urine)= colonies/mL.
1K-10K = probable contaminated sample.
10K-100K=possible UTI.
>100K= UTI.
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| Which 3 bacteria does the FDA recommend for disinfectant testing? | S.Aureus (gram +, infection of wounds), P.aeriginosa (gram -, often fatal), Solmonella typhi(gram -, hard to disinfect).
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| Given results from antibiotic testing be able to determine resistance of susceptibility to antibiotics | Measure zone of inhibition and compare to reference chart to determine resistant, intermediate, or senstive(suceptable)
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| Where does teh yeast cryptococcus cause serious infection? | Spinal fluid.
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| Define dimorphic fungi. | They exist as yeasts and molds
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| Define budding in yeast. | Yeast makes a small copy of itself that will grow into a new yeast cell.
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| How would a gram stain be used with fungi | Fungi appear gram + but larger than bacteria
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| HOw would a KOH stain be used with fungi? | KOH when mixed with tissue will digest tissue, but not fungus. A wet mount can be used to see hyphae.
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| How would an india ink stain be used with fungi? | Cryptococcus which causes meningitis can have a clear capsule.
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| What is the scotch tape test for? | Pinworm infections
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| What is the disease or condition called cysticercosis and what parasite causes it? | When humans are intermediate hosts for the pork tapeworm.
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| Infective stage and how diagnosed for giardia.. | ingest cysts. Stool sample for cysts or trophozoites
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| Infective stage and how diagnosed for Hookworm (necator americanus) | Direct skin penetration. Stool for ova.
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| Infective stage and how diagnosed for Ascaris. | Ingestion of ova. Stool for ova.
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| Infective stage and how diagnosed for beef/pork tapeworm (Taenia saginata or Solium) | Ingest larva. Stool for ova and proglottids.
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| Infective stage and how diagnosed for Trichomonas | Sexual transmission of trophozoite. Wet mount of genital secretions for trophozoite.
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| What the insect vector for Trypanosoma brucei? | Tsetse fly
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| What the insect vector for Trypanosoma cruzi | Reduviid bug
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| What the insect vector for Leismania | Sandfly
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| What the insect vector for Plasmodia? | Mosquito.
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| What two roundworms or nematodes have a larval stage migrating through the host lungs? | Ascaris lumbricoides, Necator americanus.
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| What two parasites use direct skin penetration as the way to enter the host? | Hookworms (necator americanus) and Blood fluke (Schistosoma).
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| What stage of plasmodia in the mosquito is the infective stage for humans? | Sporozoite
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| What stage of the plasmodia in the human host is the infective stage for mosquitoes? | Gamatocytes
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| What stage in the plasmodia life cycle is occuring when the human host experiences fevers and chills? | Erythrocytic cycle when merozoites are released
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| In which host does the asexual reproductive life cycle of plasmodia take place? | Human
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| In which host does the sexual reproductive life cycle of plasmodia take place? | Mosquito
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| Where in the human host does Toxoplamsa gondii infect? | CNS, heart and skel muscle.
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| In what host does teh sexual reproductive cycle of toxoplasma gondii occur? | Cats
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| What parasite causes african sleeping sickness? | Trypanosoma brucei
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| What parasite causes changas disease? | Trypanosoma cruzi
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| How would an individual get a Trichinella infection? | Ingestion of larva in skeletal muscle.
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| What have molecular techniques to identify pathogens replaced some of the traditional methods? | They've become cheaper. They can identify bacteria that doesn't grow well on media.
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| What is 16S rDNA and how is it used to identify bacteria? | A small subunit of ribosomal DNA. Species have unique 16S rDNA sequences.
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| What is PCR? | It marks a region of DNA and replicates it.
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| What are the 3 steps in PCR? | Separate the DNA strands, Annealing the primer to the template, synthesis of new DNA strands.
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| What are primers? | Small pieces of DNA that bind to specific sequences. Signals DNA polymerase to copy the region.
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| What is meant by conserved and variable regions of DNA? | Conserved=regions that are similar across species. Variable=regions are more species specific.
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| How is cycle sequencing different from PCR? | In cycle sequencing, copies are made of target DNA but each copy terminates randomly with special nucleotides.
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| What is BLAST? | A database for matching DNA sequences and ranks them based on level of similarity.
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