Lab Final Stain and Biochemical tests

How do you apply a capsule stain using Congo Red and Manevals Stain?

• put culture on one side of slide

• add congo red

• use a second slide and draw the stain across the length of the slide

• air dry slide

• cover with manevals and drain, don’t rinse

• air dru

How do you perforHow m an endospore stain with malachite green and safranin?

• smear culture on slide and dry on slide warmer

• place paper on slide

• saturate with malachite green and heat for 5 minutes by running through flame

• steam but dont boil, add more dye often

• rinse with water

• counterstain with safranin for a minute and blot dry

How do you perform a test for sugar fermentation?

• inoculate in phenol red broth w/ durham tube

• yellow = acid produced

• gas in tube = gas produced

How do you perform an indole test?

• inoculate in tryptone broth

• incubate

• add 10 drops Kovacs reagent and look for red/pink color

How do you perform an MRVP test?

• inoculate in MRVP broth and incubate

• add 5 drops methyl red indicator and swirl

• look for red

• add 10 drops KOH and 10 drops alpha-napthyol

• wait a minute and observe for red every 10 minutes

How do you perform an citrate test?

• inoculate on simmons citrate agar

• observe for blue color

How do you perform a lipid hydrolysis test?

• spot inoculate on fat agar

• observe for red

How do you perform an polysaccharide hydrolysis test?

• spot inoculate with culture

• add 30 drops of grams iodine

• look for halo

How do you perform a protein hydrolysis test?

• spot inoculate on skim milk agar

• check for clear zone

How do you perform a gelatin hydrolysis test?

• stab nu-gel tube with culture

• check for growth along stab line

• place in fridge for 30 mins, look for liquid layer on top

How do you perform a SIm test?

• stab inoculate culture ½ way down SIM agar tube

  • observe for black and spread out growth

How do you perform an LDC test?

• put culture in LDC and DC tube

• put 1cm mineral oil over both tubes

• observe for purple in LDC and yellow in DC

How do you perform a phenylalanine test?

• place 1ml of water, phen tablet, and culture in broth

• incubate

• add 3 drops fecl3 and HCl to tube

• oversve for green color within 5 minutes

How do you perform a urease test?

• place 1ml water, urease tablet in tube

• place in 80 c broth for 1 minute

• add unknown and mix

• incubate

• observe for pink color

How do you perform an ammonification test?

• inoculate culture in peptone broth

• place 50ul of inoculated broth and peptone broth in sepate wells

• add 100ul of OPA to each

• add 50 ul of TCA to each

• observe for blue/black color

How do you perform a nitrate reudction test?

• inoculate in nitrate broth

• check for n2 gas

• add sulfamic acid and check for fizzing

• add zinc dust and if fizzes, didnt reduce