BISC209: Recipes: Difference between revisions

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positive controls:  Bacillus spp, Neisseria, Micrococcus or Pseudomonas<BR><BR>
positive controls:  Bacillus spp, Neisseria, Micrococcus or Pseudomonas<BR><BR>


'''OF-glucose medium'''<BR>
0.2 per cent peptone<BR>  0.5 per cent NaCl<BR> 0.03 per cent K2HPO4<BR> 0.3 per cent agar<BR> 0.003 per cent bromthymol blue<BR> 1.0 per cent carbohydrate <BR> pH 7.1.<BR><BR>


'''special stains'''
'''special stains'''
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acid-alcohol <BR>
acid-alcohol <BR>
methylene blue</LI></UL>
methylene blue</LI></UL>
'''Role of microbes in ecosystem tests'''<BR>
'''OF-glucose medium'''<BR>
0.2 per cent peptone<BR>  0.5 per cent NaCl<BR> 0.03 per cent K2HPO4<BR> 0.3 per cent agar<BR> 0.003 per cent bromthymol blue<BR> 1.0 per cent carbohydrate <BR> pH 7.1.<BR><BR>


'''cellulose degradation medium'''<BR>
'''cellulose degradation medium'''<BR>
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or try Nutrient agar for all???<BR><BR>
or try Nutrient agar for all???<BR><BR>


'''starch medium'''<BR>
'''antibiotic plates and broths'''
3 g Beef extract<BR>
(use appropriate medium to grow isolates<BR><BR>
10 g soluble starch<BR>
12 g agar<BR>
bring up to 1 liter with distilled or deionized water<BR>
pH 7.3-7.7<BR>
 
develop with Gram Iodine<BR><BR>


'''Ammonification reagent'''<BR>
'''Ammonification reagent'''<BR>
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     NH4+ + 2[HgI4]2− + 4OH− → HgO·Hg(NH2)I + 7I− + 3H2O <BR>
     NH4+ + 2[HgI4]2− + 4OH− → HgO·Hg(NH2)I + 7I− + 3H2O <BR>
It is toxic if swallowed, inhaled or absorbed through the skin. It presents a neurological hazard and may act as a carcinogen and be a reproductive hazard. It is corrosive and causes burns.
It is toxic if swallowed, inhaled or absorbed through the skin. It presents a neurological hazard and may act as a carcinogen and be a reproductive hazard. It is corrosive and causes burns.
Wear gloves and consider performing this assay in a hood.</LI></UL><BR>
Wear gloves and consider performing this assay in a hood.</LI></UL><BR>
'''starch medium'''<BR>
3 g Beef extract<BR>
10 g soluble starch<BR>
12 g agar<BR>
bring up to 1 liter with distilled or deionized water<BR>
pH 7.3-7.7<BR>
develop with Gram Iodine<BR><BR>


Lab 6-9<BR>
Lab 6-9<BR>

Revision as of 13:47, 18 January 2010

Wellesley College-BISC 209 Microbiology -Spring 2010


Misc Recipes for test media and their reagents

Lab 4

PEA MEDIUM
  • 10g tryptose
    3g Beef extract
    5g Sodium Chloride
    2.5g Phenylethyl alcohol
    15g Agar to 1 liter distilled or deionized water
    pH 7.1-7.5.
EMB medium
  • 10g peptone
    10g Lactose
    2g dipotassium phosphate
    0.4g eosin Y
    0.065 g methylene blue
    15 g Agar
    final pH 6.9-7.3

Lab 5 (and available through lab 9
catalase test
fresh hydrogen peroxide

Oxidase test
order fresh commercial kits
positive controls: Bacillus spp, Neisseria, Micrococcus or Pseudomonas


special stains

  • Negative stain for cell morphology:
    • Nigrosin


    Endospore:
    • Malachite green hot plate Safranin
    Capsule
    • Congo red
      acid-alcohol
      fuchsin
    Acid Fast
    • Carbolfuchsin
      acid-alcohol
      methylene blue

    Role of microbes in ecosystem tests
    OF-glucose medium
    0.2 per cent peptone
    0.5 per cent NaCl
    0.03 per cent K2HPO4
    0.3 per cent agar
    0.003 per cent bromthymol blue
    1.0 per cent carbohydrate
    pH 7.1.

    cellulose degradation medium
    (use melted agar of the appropiate enrichment media choices
    or try Nutrient agar for all???

    antibiotic plates and broths (use appropriate medium to grow isolates

    Ammonification reagent
    Nesler's reagent will be purchased commercially. Please be extremely careful handling this solution.

    • Recipe: 0.09 mol/L solution of potassium tetraiodomercurate(II) (K2[HgI4]) in 2.5 mol/L potassium hydroxide. A yellow coloration indicates the presence of ammonia: at higher concentrations, a brown precipitate may form. The sensitivity as a spot test is about 0.3 μg NH3 in 2 μL.
      NH4+ + 2[HgI4]2− + 4OH− → HgO·Hg(NH2)I + 7I− + 3H2O
      It is toxic if swallowed, inhaled or absorbed through the skin. It presents a neurological hazard and may act as a carcinogen and be a reproductive hazard. It is corrosive and causes burns. Wear gloves and consider performing this assay in a hood.

    starch medium
    3 g Beef extract
    10 g soluble starch
    12 g agar
    bring up to 1 liter with distilled or deionized water
    pH 7.3-7.7

    develop with Gram Iodine

    Lab 6-9

    MRVPMethyl red test
    • dextrose peptone broth
      0.7% peptone,
      0.5% dextrose
      0.5% dipotassium phosphate
      pH 6.7-7.1
    Methyl red Reagent
    • 0.1 methyl red dye
      ethanol 00 ml
      distilled water to 500 ml
    VP Reagent A
    • 5 g alpha naphthol
      bring volume to 100 ml absolute alcohol
    VP Reagent B
    • 40g potassium hydroxide
      bring to volume with 100 ml distilled water


    SIM agar and Reagent recipes:
    • < Approximate Formula* Per Liter
      Pancreatic Digest of Casein - 20.0 g
      Peptic Digest of Animal Tissue - 6.1 g
      Ferrous Ammonium Sulfate - 0.2 g
      Sodium Thiosulfate - 0.2 g
      Agar - 3.5 g
    • Kovac's reagent: (per liter)
      p-Dimethylaminobenzaldehyde 50,0g
      Amyl Alcohol 750.0 ml
      Hydrochloric acid 250.0 ml
    • Indole Spot Test Reagent
      p-Dimethylaminocinnamaldehyde 10.0 g
      Hydrochloric acid 100.0ml
      Water, deionized 900.0 ml

    Carbohydrate fermentation medium
    This medium is available commercially

    • Purple broth base (1% peptone, 0.1% beef extract, 0.5% NaCl, 20 µg/ml bromcresol purple) plus 1% of the desired carbohydrate.
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