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Marine Agar (DSMZ Medium 123)
Composition per liter:
Agar ..................................................................15.0g
Tryptone............................................................10.0g
Peptone ...............................................................5.0g
Yeast extract ........................................................1.0g
Synthetic seawater ............................................. 1.0L
pH 7.8 ± 0.2 at 25°C
Synthetic Seawater Composition per liter:
NaCl ..................................................................24.0g
MgCl2•6H2O .....................................................11.0g
Na2SO4................................................................4.0g
CaCl2•6H2O.........................................................2.0g
KCl......................................................................0.7g
KBr......................................................................0.1g
SrCl2•6H2O .......................................................0.04g
H3BO3 ...............................................................0.03g
NaSiO3•9H2O...................................................5.0mg
NaF...................................................................3.0mg
NH4NO3 ...........................................................2.0mg
Fe3PO4•4H2O ...................................................1.0mg
Preparation of Synthetic Seawater: Add components to distilled water and bring volume to 1.0L.
Mix thoroughly.
Preparation of Medium: Add agar, tryptone, peptone, and yeast extract to synthetic seawater and
bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.8. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of Halobacillus halophilus, Halomonas spp., Vibrio harveyi,Cobetia marina, and Ruegeria atlantica.
Marine Agar 2216(DSMZ Medium 604)
Composition per liter:
NaCl ................................................................19.45g
Agar ..................................................................15.0g
MgCl2..................................................................8.8g
Peptone................................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract ........................................................1.0g
KCl....................................................................0.55g
NaHCO3 ............................................................0.16g
Ferric citrate........................................................0.1g
KBr....................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4 ..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF...................................................................2.4mg
NH4NO3 ...........................................................1.6mg
pH 7.6 ± 0.2 at 25°C
Source: This medium is available as a premixed powder from BD Diagnostic Systems.
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat while stirring and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of Hyphomonas spp., Oceanospirillum spp., Hyphomicrobium indicum, Psychroflexus gondwanensis=Flavobacterium gondwanense, Salegentibacter salegens=Flavobacterium salegens, Psychromonas antarctica, Sulfitobacter mediterraneus, Thalassomonas viridans,Vibrio spp., Marinospirillum minutulum=Oceanospirillum minutulum, Terasakiella pusilla=Oceanospirillum pusillum, Pseudoalteromonas atlantica=Alteromonas atlantica, Pseudomonas atlantica,Roseobacter spp., Erythrobacter longus, Pseudospirillum japonicum=Oceanospirillum japonicum,Marinobacter hydrocarbonoclasticus (Pseudomonas nautica), Psychrobacter spp., and Moritella japonica.
For the isolation, cultivation, and maintenance of a wide variety of heterotrophic marine bacteria.
Marine Agar with Biphenyl
Composition per liter:
NaCl ................................................................19.45g
Agar ..................................................................15.0g
MgCl2..................................................................8.8g
Peptone ...............................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract........................................................1.0g
KCl....................................................................0.55g
NaHCO3............................................................0.16g
Ferric citrate........................................................0.1g
KBr ...................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF ..................................................................2.4mg
NH4NO3 ...........................................................1.6mg
Biphenyl...........................................................1.0mg
pH 7.6 ± 0.2 at 25°C
Preparation of Medium: Add components, except biphenyl, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes. After agar solidifies, aseptically add a few crystals of biphenyl to each plate.
Use: For the cultivation and maintenance of biphenyl-utilizing marine bacteria, such as Cycloclasticus pugetii.
Marine Agar with ι-Carrageenan
Composition per 1070.0mL:
Solution A.......................................................... 1.0L
Solution B ..................................................... 60.0mL
Solution C ..................................................... 10.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per liter:
NaCl ..................................................................25.0g
Agar ..................................................................15.0g
MgSO4•7H2O......................................................5.0g
Casamino acids ...................................................2.5g
ι−Carrageenan.....................................................2.5g
NaNO3.................................................................2.0g
CaCl2•2H2O.........................................................0.2g
KCl......................................................................0.1g
Preparation of Solution A: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 100.0mL:
Na2HPO4•2H2O.................................................3.56g
Preparation of Solution B: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
FeSO4•7H2O........................................................0.3g
Preparation of Solution C: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Preparation of Medium: Aseptically add
60.0mL of sterile solution B and 10.0mL of sterile
solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and maintenance of ATCC
strain 43554.
Marine Agar
with κ- and λ-Carrageenan
Composition per 1070.0mL:
Solution A.......................................................... 1.0L
Solution B..................................................... 60.0mL
Solution C..................................................... 10.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per liter:
NaCl ..................................................................25.0g
Agar ..................................................................15.0g
MgSO4•7H2O......................................................5.0g
Casamino acids ...................................................2.5g
NaNO3 ................................................................2.0g
κ-Carrageenan...................................................1.25g
λ-Carrageenan...................................................1.25g
CaCl2•2H2O ........................................................0.2g
KCl......................................................................0.1g
Preparation of Solution A: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 100.0mL:
Na2HPO4•2H2O.................................................3.56g
Preparation of Solution B: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
FeSO4•7H2O........................................................0.3g
Preparation of Solution C: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Preparation of Medium: Aseptically add
60.0mL of sterile solution B and 10.0mL of sterile
solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation and maintenance of
Pseudomonas carrageenovora.
Marine Agar with Naphthalene
Composition per liter:
NaCl ................................................................19.45g
Agar ..................................................................15.0g
MgCl2..................................................................8.8g
Peptone ...............................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract ........................................................1.0g
KCl....................................................................0.55g
NaHCO3 ............................................................0.16g
Ferric citrate........................................................0.1g
KBr....................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4 ..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF...................................................................2.4mg
NH4NO3 ...........................................................1.6mg
Naphthalene ........................................................1mg
pH 7.6 ± 0.2 at 25°C
Preparation of Medium: Add components, except naphthalene, to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly. Gently heat
and bring to boiling. Distribute into tubes or flasks.
Autoclave for 15 min at 15 psi pressure–121°C. Pour
into sterile Petri dishes or leave in tubes. After agar
solidifies, aseptically add a few crystals of naphthalene to each plate.
Use: For the cultivation and maintenance of naphthalene-utilizing marine bacteria
Marine Agar with Sulfur
(ATCC Medium 1922)
Composition per liter:
NaCl ................................................................19.45g
Sulfur ................................................................10.0g
MgCl2..................................................................8.8g
Peptone................................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract ........................................................1.0g
KCl....................................................................0.55g
NaHCO3 ............................................................0.16g
Ferric citrate........................................................0.1g
KBr....................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4 ..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF...................................................................2.4mg
NH4NO3 ...........................................................1.6mg
pH 7.6 ± 0.2 at 25°C
Preparation of Sulfur: Autoclave sulfur for 15
min at 0 psi pressure–100°C on three successive days.
Preparation of Medium: Prepare anaerobically
under a gas phase of 80% N2 + 10% CO2 + 10% H2.
Add components, except sulfur, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling.
Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 50–55°C. Aseptically add 10.0g of sterile sulfur.
Mix thoroughly. Aseptically and anaerobically, under
a gas phase of 80% N2 + 10% CO2 + 10% H2, distribute into sterile tubes.
Use: For the cultivation and maintenance of Thermococcus litoralis.
Marine Ameba Medium
Composition per liter:
Agar ..................................................................10.0g
Malt extract .........................................................0.1g
Yeast extract........................................................0.1g
Artificial seawater.............................................. 1.0L
Artificial Seawater:
Composition per liter:
NaCl ..................................................................27.5g
MgSO4 •7H2O...................................................6.78g
MgCl2•6H2O .....................................................5.38g
KCl....................................................................0.72g
NaHCO3............................................................. 0.2g
CaCL2•2H2O.......................................................1.4g
Preparation of Artificial Seawater: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Add components to artificial seawater and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute
into tubes or flasks. Autoclave for 15 min at 15 psi
pressure–121°C. Pour into sterile Petri dishes or
leave in tubes.
Use: For the cultivation of Cochliopodium clarum,
Heteramoeba clara, Ling***oeba leei, Paramoeba
pemaquidensis, and Vannella species.
Marine Broth 2216
(LMG Medium 164)
Composition per liter:
NaCl ................................................................19.45g
MgCl2..................................................................8.8g
Peptone ...............................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract........................................................1.0g
KCl....................................................................0.55g
NaHCO3............................................................0.16g
Ferric citrate........................................................0.1g
KBr ...................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF...................................................................2.4mg
NH4NO3 ...........................................................1.6mg
pH 7.6 ± 0.2 at 25°C
Source: This medium is available as a premixed
powder from BD Diagnostic Systems.
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat while stirring and bring
to boiling. Distribute into tubes or flasks. Autoclave
for 15 min at 15 psi pressure–121°C.
Use: For the cultivation of Vibrio liquefaciens and
for the isolation, cultivation, and maintenance of a
wide variety of heterotrophic marine bacteria.
Marine Broth with Biphenyl
Composition per liter:
NaCl ................................................................19.45g
MgCl2..................................................................8.8g
Peptone................................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract ........................................................1.0g
KCl....................................................................0.55g
NaHCO3 ............................................................0.16g
Ferric citrate........................................................0.1g
KBr....................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4 ..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF...................................................................2.4mg
NH4NO3 ...........................................................1.6mg
Biphenyl ...........................................................1.0mg
pH 7.6 ± 0.2 at 25°C
Preparation of Medium: Add components, except biphenyl, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Distribute into
tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add a few crytals of biphenyl to each tube or flask.
Use: For the cultivation of biphenyl-utilizing marine
bacteria.
Marine Broth with ι-Carrageenan
Composition per 1070.0mL:
Solution A.......................................................... 1.0L
Solution B ..................................................... 60.0mL
Solution C ..................................................... 10.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per liter:
NaCl ..................................................................25.0g
MgSO4•7H2O......................................................5.0g
Casamino acids ...................................................2.5g
ι-Carrageenan .....................................................2.5g
NaNO3 ................................................................2.0g
CaCl2•2H2O ........................................................0.2g
KCl......................................................................0.1g
Preparation of Solution A: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 100.0mL:
Na2HPO4•2H2O.................................................3.56g
Preparation of Solution B: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
FeSO4•7H2O........................................................0.3g
Preparation of Solution C: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Preparation of Medium: Aseptically add
60.0mL of sterile solution B and 10.0mL of sterile
solution C to 1.0L of sterile solution A. Mix thoroughly. Pour into sterile Petri dishes or distribute into
sterile tubes.
Use: For the cultivation of ATCC strain 43554.
Marine Broth
with κ- and λ-Carrageenan
Composition per 1070.0mL:
Solution A.......................................................... 1.0L
Solution B..................................................... 60.0mL
Solution C..................................................... 10.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per liter:
NaCl ..................................................................25.0g
MgSO4•7H2O......................................................5.0g
Casamino acids ...................................................2.5g
NaNO3 ................................................................2.0g
κ-Carrageenan...................................................1.25g
λ-Carrageenan...................................................1.25g
CaCl2•2H2O ........................................................0.2g
KCl......................................................................0.1g
Preparation of Solution A: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure–121°C.
Solution B:
Composition per 100.0mL:
Na2HPO4•2H2O.................................................3.56g
Preparation of Solution B: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
FeSO4•7H2O........................................................0.3g
Preparation of Solution C: Add component to
distilled/deionized water and bring volume to
100.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C.
Preparation of Medium: Aseptically add
60.0mL of sterile solution B and 10.0mL of sterile
solution C to 1.0L of sterile solution A. Mix thoroughly. Distribute into sterile tubes or flasks.
Use: For the cultivation and maintenance of
Pseudomonas carrageenovora.
Marine Broth with Naphthalene
Composition per liter:
NaCl ................................................................19.45g
MgCl2..................................................................8.8g
Peptone................................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract ........................................................1.0g
KCl....................................................................0.55g
NaHCO3 ............................................................0.16g
Ferric citrate........................................................0.1g
KBr....................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4 ..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF...................................................................2.4mg
NH4NO3 ...........................................................1.6mg
Naphthalene ........................................................1mg
pH 7.6 ± 0.2 at 25°C
Preparation of Medium: Add components, except biphenyl, to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Distribute into
tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add a few crytals of naphthalene to each tube or flask.
Use: For the cultivation of naphthalene-utilizing
marine bacteria.
Marine Broth with Sulfur
Composition per liter:
NaCl ................................................................19.45g
Sulfur ................................................................10.0g
MgCl2..................................................................8.8g
Peptone ...............................................................5.0g
Na2SO3..............................................................3.24g
CaCl2...................................................................1.8g
Yeast extract........................................................1.0g
KCl....................................................................0.55g
NaHCO3............................................................0.16g
Ferric citrate........................................................0.1g
KBr ...................................................................0.08g
SrCl2..................................................................0.03g
H3BO3 ...............................................................0.02g
Na2HPO4..........................................................8.0mg
Na2SiO3............................................................4.0mg
NaF ..................................................................2.4mg
NH4NO3 ...........................................................1.6mg
pH 7.6 ± 0.2 at 25°C
Preparation of Sulfur: Autoclave for 15 min at 0
psi pressure–100°C on three successive days.
Preparation of Medium: Prepare anaerobically
under a gas phase of 80% N2 + 10% CO2 + 10% H2.
Add components, except sulfur, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling.
Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 50°C. Aseptically add 10.0g of sulfur. Mix thoroughly. Aseptically and anaerobically, under a gas
phase of 80% N2 + 10% CO2 + 10% H2, distribute
into sterile tubes.
Use: For the cultivation of Thermococcus litoralis.
Marine Caulobacter Medium
Composition per liter:
Proteose peptone...............................................10.0g
Yeast extract........................................................3.0g
Artificial seawater.............................................. 1.0L
pH 7.2–7.4 at 25°C
Artificial Seawater:
Composition per liter:
Commercially available marine
aquarium salts mixture ...........................variable
Preparation of Artificial Seawater: Add commercially available marine aquarium salts mixture to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly.
Preparation of Medium: Combine components.
Mix thoroughly. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure–121°C.
Use: For the cultivation of Caulobacter halobacteroides and Caulobacter maris.
Marine Chlorobiaceae Medium 2
Composition per 1051.0mL:
Solution 1.................................................... 950.0mL
Na2S•9H2O solution...................................... 60.0mL
NaHCO3 solution .......................................... 40.0mL
Vitamin B12 solution ....................................... 1.0mL
pH 6.8 ± 0.2 at 25°C
Solution 1:
Composition per 950.0mL:
NaCl ..................................................................20.0g
MgSO4•7H2O......................................................3.0g
KH2PO4...............................................................1.0g
NH4Cl .................................................................0.5g
CaCl2•2H2O.......................................................0.05g
Trace elements solution SL-8 ......................... 1.0mL
Preparation of Solution 1: Add components to
distilled/deionized water and bring volume to
950.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 45°–50°C.
Trace Elements Solution SL-8:
Composition per liter:
Disodium EDTA .................................................5.2g
FeCl2•4H2O.........................................................1.5g
CoCl2•6H2O ......................................................0.19g
MnCl2•4H2O .......................................................0.1g
ZnCl2.................................................................0.07g
H3BO3 ...............................................................0.06g
NaMoO4•2H2O..................................................0.04g
CuCl2•2H2O ......................................................0.02g
NiCl2•6H20........................................................0.02g
Preparation of Trace Elements Solution SL8: Add components to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly.
Na2S•9H2O Solution:
Composition per 100.0mL:
Na2S•9H2O..........................................................5.0g
Preparation of Na2S•9H2O Solution: Add
Na2S•9H2O to distilled/deionized water and bring
volume to 100.0mL. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 45°–50°C.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ..............................................................5.0g
Preparation of NaHCO3 Solution: Add
NaHCO3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Vitamin B12 Solution:
Composition per 100.0mL:
Vitamin B12 ......................................................2.0mg
Preparation of Vitamin B12 Solution: Add vitamin B12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: To 950.0mL of cooled,
sterile solution 1, aseptically add 60.0mL of sterile
Na2S•9H2O solution, 40.0mL of sterile NaHCO3 solution, and 1.0mL of sterile vitamin B12 solution. Mix
thoroughly. Adjust pH to 6.8 with sterile H2SO4 or
Na2CO3
. Aseptically distribute into sterile 50.0mL
or 100.0mL bottles with metal screw-caps and rubber
seals. Completely fill bottles with medium except for
a pea-sized air bubble.
Use: For the isolation and cultivation of marine
members of the Chlorobiaceae.
Marine Chromatiaceae Medium 2
Composition per 1051.0mL:
Solution 1.................................................... 950.0mL
Na2S•9H2O solution...................................... 60.0mL
NaHCO3 solution.......................................... 40.0mL
Vitamin B12 solution ....................................... 1.0mL
pH 7.3 ± 0.2 at 25°C
Solution 1:
Composition per 950.0mL:
NaCl ..................................................................20.0g
MgSO4•7H2O......................................................3.0g
KH2PO4...............................................................1.0g
NH4Cl .................................................................0.5g
CaCl2•2H2O ......................................................0.05g
Trace elements solution SL-8 ......................... 1.0mL
Preparation of Solution 1: Add components to
distilled/deionized water and bring volume to
950.0mL. Mix thoroughly. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 45°–50°C.
Trace Elements Solution SL-8:
Composition per liter:
Disodium EDTA.................................................5.2g
FeCl2•4H2O.........................................................1.5g
CoCl2•6H2O ......................................................0.19g
MnCl2•4H2O .......................................................0.1g
ZnCl2.................................................................0.07g
H3BO3 ...............................................................0.06g
NaMoO4•2H2O..................................................0.04g
CuCl2•2H2O ......................................................0.02g
NiCl2•6H20........................................................0.02g
Preparation of Trace Elements Solution SL8: Add components to distilled/deionized water and
bring volume to 1.0L. Mix thoroughly.
Na2S•9H2O Solution:
Composition per 100.0mL:
Na2S•9H2O..........................................................5.0g
Preparation of Na2S•9H2O Solution: Add
Na2S•9H2O to distilled/deionized water and bring
volume to 100.0mL. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 45°–50°C.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ..............................................................5.0g
Preparation of NaHCO3 Solution: Add
NaHCO3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Vitamin B12 Solution:
Composition per 100.0mL:
Vitamin B12 ......................................................2.0mg
Preparation of Vitamin B12 Solution: Add vitamin B12 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: To 950.0mL of cooled,
sterile solution 1, aseptically add 60.0mL of sterile
Na2S•9H2O solution, 40.0mL of sterile NaHCO3 solution, and 1.0mL of sterile vitamin B12 solution. Mix
thoroughly. Adjust pH to 7.3 with sterile H2SO4 or
Na2CO3
. Aseptically distribute into sterile 50.0mL
or 100.0mL bottles with metal screw-caps and rubber
seals. Completely fill bottles with medium except for
a pea-sized air bubble.
Use: For the isolation and cultivation of marine
members of the Chromatiaceae.
Marine Cytophaga Agar
Composition per liter:
Agar ..................................................................15.0g
Nutrient broth......................................................8.0g
Yeast extract ........................................................5.0g
Salt solution ....................................................... 1.0L
Salt Solution:
Composition per liter:
NaCl ................................................................12.86g
MgCl2................................................................2.48g
KCl....................................................................0.75g
CaCl2.................................................................0.56g
Fe(SO4)2(NH4)2...............................................0.048g
Preparation of Salt Solution: Add components
to distilled/deionized water and bring volume to
1.0L. Mix thoroughly.
Preparation of Medium: Add solid components
to 1.0L of salt solution. Mix thoroughly. Gently heat
while stirring and bring to boiling. Distribute into tubes
or flasks. Autoclave for 15 min at 15 psi pressure–
121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of Cytophaga species.
Marine Cytophaga Medium
Composition per liter:
NaCl ..................................................................24.7g
Agar ..................................................................15.0g
MgSO4•7H2O......................................................6.3g
MgCl2•6H2O .......................................................4.6g
Tryptic digest of casein.......................................1.0g
Yeast extract........................................................1.0g
KCl......................................................................0.7g
NaHCO3
solution .......................................... 10.0mL
CaCl2•2H2O solution..................................... 10.0mL
pH 7.2 ± 0.2 at 25°C
NaHCO3 Solution:
Composition per 10.0mL:
NaHCO3..............................................................0.2g
Preparation of NaHCO3 Solution: Add
NaHCO3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C.
CaCl2•2H2O Solution:
Composition per 10.0mL:
CaCl2•2H2O ........................................................1.2g
Preparation of CaCl2•2H2O Solution: Add
CaCl2•2H2O to distilled/deionized water and bring
volume to 10.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except NaHCO3 solution and CaCl2•2H2O solution, to
distilled/deionized water and bring volume to
980.0mL. Mix thoroughly. Gently heat and bring to
boiling. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 50°–55°C. Aseptically add 10.0mL
of sterile NaHCO3 solution and 10.0mL of sterile
CaCl2•2H2O solution. Mix thoroughly. Pour into
sterile Petri dishes or distribute into sterile tubes.
Use: For the cultivation of Cytophaga species, Flexibacter species, Microscilla species, and Saprospira
grandis.
Marine Cytophaga Medium A
Composition per liter:
Agar ..................................................................15.0g
Pancreatic digest of casein..................................2.0g
Beef extract .........................................................0.5g
Yeast extract........................................................0.5g
Sodium acetate....................................................0.2g
Seawater...................................................... 700.0mL
pH 7.2 ± 0.2 at 25°C
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation of Flexibacter maritimus.
Marine Cytophaga Medium B
Composition per liter:
Agar ..................................................................15.0g
Pancreatic digest of casein..................................2.0g
Beef extract .........................................................0.5g
Yeast extract ........................................................0.5g
Sodium acetate....................................................0.2g
Seawater...................................................... 500.0mL
pH 7.2 ± 0.2 at 25°C
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation of Vibrio ordalii.
Marine Cytophaga Medium C
Composition per liter:
Agar ..................................................................15.0g
Pancreatic digest of casein..................................2.0g
Beef extract .........................................................0.5g
Yeast extract ........................................................0.5g
Sodium acetate....................................................0.2g
pH 7.2 ± 0.2 at 25°C
Preparation of Medium: Add components to
seawater and bring volume to 1.0L. Mix thoroughly.
Gently heat and bring to boiling. Distribute into tubes
or flasks. Autoclave for 15 min at 15 psi pressure–
121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation of Cytophaga agarovorans,
Cytophaga fermentans, and Cytophaga salmonicolor.
Marine Desulfovibrio Medium
Composition per liter:
Solution A................................................... 980.0mL
Solution B ..................................................... 10.0mL
Solution C ..................................................... 10.0mL
pH 7.8 ± 0.2 at 25°C
Solution A:
Composition per 980.0mL:
NaCl ..................................................................25.0g
DL-Sodium lactate...............................................2.0g
MgSO4•7H2O......................................................2.0g
Na2SO4................................................................1.0g
NH4Cl .................................................................1.0g
Yeast extract........................................................1.0g
K2HPO4...............................................................0.5g
CaCl2•2H2O ........................................................0.1g
Resazurin .........................................................1.0mg
Preparation of Solution A: Add components to
distilled/deionized water and bring volume to
980.0mL. Mix thoroughly. Gently heat and bring to
boiling. Continue boiling for 3-4 min. Allow to cool
to room temperature while gassing under 100% N2.
Solution B:
Composition per 10.0mL:
FeSO4•7H2O........................................................0.5g
Preparation of Solution B: Add FeSO4•7H2O to
distilled/deionized water and bring volume to
10.0mL. Mix thoroughly.
Solution C:
Composition per 10.0mL:
Ascorbic acid ......................................................0.1g
Sodium thioglycolate ..........................................0.1g
Preparation of Solution C: Add components to
distilled/deionized water and bring volume to
10.0mL. Mix thoroughly.
Preparation of Medium: To 980.0mL of cooled
solution A, anaerobically add 10.0mL of solution B
and 10.0mL of solution C. Mix thoroughly. Adjust
pH to 7.8 with NaOH. Distribute into tubes or flasks.
During distribution, swirl the medium to keep the
precipitate in suspension. Autoclave for 15 min at 15
psi pressure–121°C.
Use: For the cultivation and maintenance of Desulfovibrio desulfuricans, Desulfovibrio salexigens,
and Desulfovibrio vulgaris.
Marine Flagellate Medium
Composition per 15.0mL:
Rice grains ..........................................................2.0g
Seawater........................................................ 15.0mL
Preparation of Medium: Autoclave rice grains
for 15 min at 15 psi pressure–121°C. Add 2.0g of
sterile rice grains to 15.0mL of filter-sterilized seawater. Aseptically distribute into T-25 tissue culture
flasks.
Use: For the cultivation of Acanthoecopsis unguiculata, Amastigomonas species, Bicosoeca vacillans,
Bodo designis, Bodo variabilis, Caecitellus parvulus,
Choanoeca perplexa, Codosiga gracilis, Diaphanoeca
grandis, Entosiphon species, Goniomonas species, Procryptobia species, Pseudobodo tremulans, Rhynchomonas nasuta, Salpingoeca urceolata, Stephanoeca diplocostata, and Stephanopogon apogon.
Marine Flagellate Medium
with B-Vitamins
Composition per liter:
Seawater...................................................... 990.0mL
Vitamin solution............................................ 10.0mL
Vitamin Solution:
Composition per 100.0mL:
Thiamine•HCl ...................................................0.15g
Calcium D-(+)-pantothenate..............................0.05g
Nicotinamide.....................................................0.05g
Pyridoxal•HCl ...................................................0.05g
Riboflavin .........................................................0.05g
Folic acid.........................................................0.025g
Pyridoxamine•HCl ..........................................0.025g
Biotin .............................................................12.5mg
Preparation of Vitamin Solution: Add components to distilled/deionized water and bring volume
to 100.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Allow natural seawater
to age for 2 months. Filter sterilize. Aseptically add
100.0mL of sterile vitamin solution. Mix thoroughly.
Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Oikomonas species.
Marine Glucose Trypticase™
Yeast Extract Agar
(MGTY Agar)
Composition per liter:
Agar ....................................................................8.0g
Glucose ...............................................................2.0g
Pancreatic digest of casein..................................1.0g
Yeast extract ........................................................1.0g
L-Cysteine•HCl•H2O............................................0.5g
Seawater...................................................... 750.0mL
Tris-HCl buffer (5.0 mM, pH 7.5) ................ 50.0mL
Resazurin (0.1% solution)............................... 1.0mL
pH 7.5 ± 0.2 at 25°C
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat while stirring and bring
to boiling. Distribute into tubes or flasks under 97%
N2 + 3% H2. Cap with rubber stoppers and place
tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust.
Use: For the cultivation and maintenance of Spirochaeta isovalerica.
Marine Glucose Trypticase™
Yeast Extract Broth
(MGTY Broth)
Composition per liter:
Glucose ...............................................................2.0g
Pancreatic digest of casein..................................1.0g
Yeast extract........................................................1.0g
L-Cysteine•HCl•H2O............................................0.5g
Seawater...................................................... 750.0mL
Tris-HCl buffer (5.0 mM, pH 7.5) ................ 50.0mL
Resazurin (0.1% solution) .............................. 1.0mL
pH 7.5 ± 0.2 at 25°C
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat while stirring and bring
to boiling. Distribute into tubes or flasks under 97%
N2 + 3% H2. Cap with rubber stoppers and place
tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust.
Use: For the cultivation and maintenance of Spirochaeta isovalerica.
Marine Methanogenium
Alcohol Medium
Composition per 1003.0mL:
NaCl ..................................................................21.0g
MgCl2•6H2O .......................................................3.0g
NaCl ....................................................................1.0g
KCl......................................................................0.5g
MgCl2•6H2O .......................................................0.5g
NH4Cl .................................................................0.4g
Sodium acetate•3H2O..........................................0.4g
KH2PO4...............................................................0.2g
CaCl2•2H2O ........................................................0.1g
NaHCO3 solution.......................................... 60.0mL
2-Propanol....................................................... 5.0mL
Na2S•9H2O solution........................................ 3.0mL
Cyanocobalamin solution ............................... 1.0mL
Selenite-molybdate-tungstate solution............ 1.0mL
Thiamine solution ........................................... 1.0mL
Trace elements solution .................................. 1.0mL
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