Micro Practical 2 – Flashcards

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Bergey's Manual of Determinative Bacteriology
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Journal to find unknown
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Unknown Classification
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Phylum Gammaproteobacteria | Family Enterobacteriaceae
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Oxygen Tests
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Thioglycollate Broth, Candle Jar, GlassPak
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Obligate Aerobes - Thioglycollate
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Bacteria at top of both
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Facultative Anaerobes - Thioglycollate
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Bacteria growth throughout, greater growth at top (oxygen)
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Obligate Anaerobe - Thioglycollate
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Only growth at bottom of tube
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Microaerophile - Thioglycollate
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Growth in middle of tube
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Candle Jar
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Grows microaerophiles - 3-5% CO2 and 8-10% O2
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GasPak
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Grows anaerobes - removes oxygen
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Thioglycollate Broth
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Reduces oxygen, does not remove all oxygen. Resazurin - Indicator (oxygen = pink)
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Catalase Test
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Catalase (H2O2 --> O2 + H20)
+ = bubbles
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Oxidase test
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- OX+ must use O2 as final electron acceptor
- Differentiates between Enterobacteriaceae (-) and Pseudomonadaceae (+)
- Cytochrome oxidase (reduction of oxygen in ETC)
+ = blue / purple
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oxidase positive - purple / blue
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Characteristics of Enterobacteriaceae
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- Gram - 

- Cocco bacilli

- facultative anaerobes

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IMViC Test
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Indole, Methyl Red, Voges-Proskauer, Citrate, H2S production, motility
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Indole Tests for
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Tryptophan --> Indole (by tryptophanase)
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Indole Media
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SIM Agar Deep
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Indole +
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hot pink / red
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Indole Reagent
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Kovacs
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H2S Test
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Sim Agar Deep, can see motility if positive. Positive = black
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TTC motility deep
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red = positive, tetrazolium chloride
strict aerobes do not grow well
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Simmons Citrate tests for
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Citrate Utilization
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Simmons Citrate Positive
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Blue
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Methyl Red Media
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MRVP Broth
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Methyl Red purpose
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use of glucose with acidic end product
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Voges Proskauer Test
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Determines use of glucose - tests for acetoin as end product
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Simmons Citrate Purpose
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Is citrate sole carbon source
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Simmons Citrate Indicator
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brom thymol blue
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Carbohydrate Utilization Media
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Phenol Red Sugar Broth w/ durham tube (lactose, mannitol, glucose, sucrose)
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Citrate
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Indole
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Methyl Red
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Phenol Red
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VP
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Methyl Red Positive
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Red
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Voges Proskauer Positive
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Red
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Nitrogen Fixation
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N2 --> Ammonium or NO2

necessary to synthesize amino acids or nucleic acids
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Nitrification
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Ammonia --> nitrite
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Denitrification
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Nitrate to Nitrite to Nitric Oxide to Nitrous Oxide to N2
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Nitrate Reduction Test
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tests for production of nitrate reductase (reduces nitrate to nitrite)
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Nitrate Reduction Media
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Nitrate Broth
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Nitrate Reduction Reagent
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Reagent A and B
(optional: zinc)
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Nitrate Reduction +
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red or yellow
(- is colorless or pink with zinc)
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Decarboxylation Test
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Tests for decarboxylase - breaks carboxyl off of amino acid
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Decarboxylase media
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arginine decarboxylase, ornithine decarboxylase, lysine decarboxylase
with mineral oil (needs anaerobic condition)
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Positive decarboxylase
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purple
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Deamination Test
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Removes amino group from amino acid
Phenylaline deaminates to phenylpyruvic acid
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Deamination Media
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Phenylaline slant
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Gelatin Hydrolysis
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Gelitnase turns gelatin to liquid = positive = mushy goop
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[image]
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Deaminase
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Decarboxylation
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Nitrate negative with zinc
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Nitrate Reduction
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Casein Hydrolysis
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Caseinase breaks down casein into amino acids
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Casein media
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skim milk agar
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Casein positive
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white to clear (zone of clearing)
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Casein enzyme
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Caseinase
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Starch hydrolysis
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Amylase hydrolyzes starch into smaller sugars
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Starch hydrolysis enzyme
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amylase
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Starch hydrolysis media
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Starch afar plate
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Starch hydrolysis reagent
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Gram's iodine
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Starch hydrolysis positive
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positive = yellow / clear zones
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Lipid hydrolysis
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lipase breaks down tributyrin into fatty acids
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Lipid hydrolysis enzyme
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Lipase
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Lipid hydrolysis Media
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Tributyrin agar plate with methlene blue indicator
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Urea Hydrolysis
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Urease breaks down ammonia and carbon dioxide
- Determines if urea can be used as energy source
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Urea Hydrolysis Indicator
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Phenol red
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Urea hydrolysis +
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hot pink
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Casein
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Lipid Hydrolysis
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Starch hydrolysis
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Urea Hydrolysis
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API 20E
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Gram negative enteric rods, dehydrated tests in each compartments. make code and determine name of bacteria
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Staph samples from
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nasal swab
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strep samples from
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throat swab
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Staph characteristics
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Gram +, microbiota of skin and nasal cavity, facultative anaerobe, haloduric
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Mannitol Salt Agar
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High Sodium Chloride - selective
Mannitol fermentation - differential
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MSA mannitol fermentation indicator and positive
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Sugar fermentation (acid production) phenol red indicator red to yellow
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Strep characteristics
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Gram +, catalase -, facultative anaerobe, prefer high co2, microbiota of skin, upper respiritory and GI
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Alpha hemplysis
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partial digestion
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beta hemolysis
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complete digestion
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gamma hydrolysis
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no digestion
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S pneumoniae
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S pyogenes
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[image]
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Hydrolysis
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Mannitol
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Columbia Naladixic Agar (CNA)
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- Naladixic Acid - Selective, Antibiotic - only G+ Grow
- Differential - 5% sheep blood
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Urine in Kidney
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Sterile (exception - nephritis, kidney infection)
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UTI cell count
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over 100k cells / ml
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EMB (eosin Methylene Blue) agar
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- Selective - methylene blye inhibits G+ bacteria
- Differential - Lactose - Lactose fermenting bacteria produce acid (ie E Coli)
- Eosin indicator changes to purple with green metallic, non lactose fermenter - brown of pink
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Cells/ml count
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# colonies on plate * 100
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EMB Examples
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[image]
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EMB Examples
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Hemolysis
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Staph catalase result
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+
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Staph hemolysis
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either beta or gamma
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SM110
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Staph test - pathogenic are orange or yellow
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DNAse
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exoenzyme that breaks down DNA
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bile esculin
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- strep test - bile inhibits growth of all but enterococci and strep group D
- Use of esculin sugar (ifbacteria is able to grow)
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NaCL broth (strep test)
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Only Enterococci can grow
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S Aureus virulence factors
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Pyogenic Toxin - pus
Enterotoxin - food poisoning
Cytotoxins
Toxic Shock Toxins
Coagulase - clotting
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Staph catalase result
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+
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Strep catalase result
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-
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Coagulase
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exoenzyme for clotting, only in S aureus
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Strep pyogenes diseases
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skin infections, strep throat, nectotizing fascitis
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Strep agalactiae
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Group B strep - neonatal sepsis
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Strep pneumonia
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pneumonia
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Enterococci
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recently separated from strep, highly antibiotic resistance, nosocomial infections
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Strep virulence factors
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- hyaluronidase - breaks down hyaluronic acid b/w cells
- Leucidin - kill WBCs
- Streptolysin - hemolytic toxin
- scarlet fever toxin
- TSS
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Strep NaCl test
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turbid = enterococci
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Antigen
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any feature that can elicit an immune response
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Antibodies
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(Immunoglobulins) produced by immune response, bind to antigens
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Agglutination
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antibodies clump to microorganisms in response to antigens
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Monospot test purpose
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- testing for antibody
- Ebstein Barr viral antigen
- EBV causes mono, burkins lymphoma
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Monospot test process
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Latex particles coated in mono antigen from cattle. Agglutination = positive (patient has antibodies)
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Test for antigen with known antibody (3)
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- agar, then perform biochemical tests, takes 1-2 days
- Difco Staph Latex kit - latex conjugated antibody
- Clearview Exact Strep Dipstick - rapid strep test
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Difco Staph Latex
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- can confirm S Aureus ID
- mix blue latex coated with antibody with patient sample
- A aureus = agglutination
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Clearview Exact Strep Dipstick
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- rapid strep test, directly from throat swab
- positive = 2 lines
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Bile esculin
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[image]
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DNAse
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[image]
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MRSA
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[image]
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Staph aureus
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Bacteriophages
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viruses that only infect bacteria
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Lytic infection
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causes death of bacteria
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plaque
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spot on bacterial lawn where all bacteria have been killed
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Phage typing
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can be used to ID strains of bacteria
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Coliphages
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bacteriophages that infect different strains of bacteria
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PFU / mL (plaque forming units per mL)
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plaques / (dilution of tube x amount plated)
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Monospot results - right = positive
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[image]
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Monospot Test
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Rapid Stress Test
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Arthropods
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Invertebrates (insects and arachnids)
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Ectoparasites
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Arthropod vector of disease when living on human body
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Class Insecta
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- Wings or no wings, 3 pairs legs, antennae
- body parts - head, thorax, abdomen
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Class Arachnid
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No wings, 4 pairs legs, no antennae
- oophalothorax and abdomen
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Scarcoptes
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Itch mite
- mange, scabies, STI
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Ixodes
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Hard body tick - lyme disease
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Dermacentor
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Hard Body Tick - Tularemia, Rocky Mountain Spotted Fever
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Ornithodoros
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Soft Body Tick
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Pediculus
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Body Louse - typhus
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Pithirus
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Pubic Louse - STD, crabs
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Anopheles
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Mosquito - Malaria
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Aedes
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Mosquito - dengue fever, viral encephalitis
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Pulex
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Flea
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Helminth Characteristics
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Eukaryotic, multicellular
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Cestodes
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- Tapeworms
- Phylum Platyhelminths
- Flat
- Proglottid segments w/ reproductive organs
- Hermaphrodite
- Scolex head w/ suckers or hooks
- Taenia
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Trematodes (flukes)
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- Platyhelminths phylum
- flat, leaf shaped
- unsegmented
- suckers
- 2 sexes
- Schistosoma species
- Fasciola hepatica
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Nematode
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- Phylum Nematoda
- Roundworms
- 2 sexes ; eggs in body
- unsegmented
- cutting teeth
- tail of male vs female
- encystment (trichinella)
- Enterobius, Ancylostoma, Trichinella, Dirofilaria
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Aedes Mosquito
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Ancylostoma
(Nematode)
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Arachnid body parts
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Dermacentor - hard body tick
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[image]
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Dirofilaria - nematode, dog heart worm
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Enterobius, nematode (pinworm)
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Fasciola, trematode (platyhelminths)
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Insect Body Parts
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Ixodes - hard tick
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[image]
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Ornithodorus - soft body tick
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[image]
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Proglottids -cestodes (platyhelminths) tapeworms
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[image]
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pulex - flea
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Schistoma - trematodes (platyhelminths)
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[image]
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Taenia - cestodes (platyhelminths)
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[image]
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Trichinella - nematode
- encystment
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[image]
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Pediculous - Louse
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Gaspak strip indicator
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methylene blue
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Tick and mite class
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Arachnida
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Louse and flea class
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Insecta
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Platyhelminths (phylum) Classes
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Cestode (tapeworms) and trematode (flukes)
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Nematodes
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Roundworms, seperate sexes, more evolved than platyhelminths
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decarboxylase medium
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moeller broth, no reagent
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phenylaline reagent (deaminase)
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ferric acid
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decarboxylase indicator
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brom cresol purple
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Staph pathogens
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S aureus (most problematic), S epidermidis, S saprophyticus, S haemolyticus, S hominis
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Staph culturing media
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blood agar - good for fastidious bacteria
CNA - has antibiotic that inhibits G-
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Staph ID tests
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nasal swabs
MSA (salt tolerance)
CNA (sensitivity to novobicin and hemolysis)
Catalase +
SM110
DNAse
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Strep pathogens
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Streo pyogenes, strep agalctiae ,strep pneumoniae
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Strep tests
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throat swab
blood agar (incubate in candle jar)
CNA plate
bile esculin
NaCl - salt resistance
SXT (antibiotic)
Catalase -
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urea indicator
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phenol red
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H2S Indicator
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Sodium thiosulfate
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VP Reagents
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Barritts A & B (light pink/red = positive)
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Lipid hydrolysis media
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methylene blue agar
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Starch Hydrolysis Media
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Starch agar
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