Trichosphaerium sp. (ATCC® 40319)

Strain Designations: Am-I-7PL (Amoeba-I-7 Plastic)  /  Depositor: M Polne-Fuller  /  Biosafety Level: 1

Permits and Restrictions

View Permits

Strain Designations Am-I-7PL (Amoeba-I-7 Plastic)
Application
degrades plastics
microorganisms for degrading plant cell walls
Biosafety Level 1
Isolation
mutant derived from ATCC 40318, Santa Barbara Co., CA, ?
Product Format test tube
Type Strain no
Medium Medium 1405: HESNW medium
Growth Conditions
Temperature: 20.0°C
Protocol: ATCCNO: 40318 SPEC: This strain is shipped as a test tube culture (10 ml of culture in a 20 x 125-mm screw-capped test tube). Upon arrival, rub the internal surfaces of the tube with a sterile cotton swab to dislodge any adhering amoebae. Aseptically divide the entire contents into 2 equal portions and distribute to T-25 tissue culture flasks. Add 5 ml of fresh ATCC medium 1405 and 0.5 ml of heat-killed Dunaliella tertiolecta ATCC 30929 to each flask. Screw the caps on tightly and incubate at 20-25C. Transfer every 4-6 weeks. Rub the surface of the flask to be subcultured with a sterile cotton swab, agitate and aseptically transfer a 0.25-0.5 ml aliquot to a fresh flask containing 10 ml of ATCC medium 1405 supplemented with 0.5 ml of heat-killed algal suspension. Incubate as above. Note: Since this strain is cultivated with heat-killed Dunaliella tertiolecta ATCC 30929 as a food source, the food should be prepared before the strain is ordered. The food is prepared as follows: 1. Establish a culture of ATCC 30929 in 5 ml of ATCC medium 1194 broth in a 16 x 125-mm screw-capped test tube. Incubate with the cap on loose under 50-75 (Einsteins/m2/s of light at 25C. 2. When the culture reaches early stationary phase, aseptically transfer 0.25-ml aliquots to each of 10 250-ml cotton-plugged Erlenmeyer flasks containing 100 ml of ATCC medium 1194 broth. Incubate as above. 3. When the cultures reach early stationary phase (approximately 10-14 days), harvest as follows: Aseptically transfer algal suspensions to 50-ml plastic centrifuge tubes. Centrifuge at 400-500 x g for 5 minutes. 4. Decant supernatant and resuspend each pellet in approximately one ml of ATCC medium 1405. Pool all suspensions in a single centrifuge tube and centrifuge as in step 3. The cell pellet should be approximately 5 ml. 5. Resuspend the cell pellet to a final volume of approximately 14 ml with ATCC medium 1405 and transfer to a 15-ml plastic centrifuge tube. Centrifuge as in step 3. Discard the supernatant. 6. Resuspend the cell pellet to a final volume of approximately 14 ml with ATCC medium 1405 and centrifuge as in step 3. Discard the supernatant. 7. Resuspend the final cell pellet to a final volume of 10 ml with ATCC medium 1405. Transfer the cell suspension to a sterile 125-ml screw-capped bottle and aseptically add 40 ml of ATCC medium 1405 bringing the final volume to 50 ml.WARN: Last ATCC#: 40318, unprocessed line: 8. Place bottle prepared in step 7 in a 60C water bath to a level such that the liquid level of the water bath is above that of the suspension in the bottle. Incubate for a total of 30 minutes, swirling the bottle at 10-minute intervals. Allow the bottle to cool to room temperature. This treatment will kill all algal cells. 9. As a check for viable cells, add 3 drops of the cell suspension prepared in step 10 to the edge of a 100-mm petri plate containing ATCC medium 1194 agar. Hold the plate vertically to allow the drops to move to the opposite edge. Incubate plate at 25C under 50-75 uE of light for 7 days. 10. The heat-killed algae can be stored at 4C for up to one year.
Subcultivation
Protocol: ATCCNO: 40318 SPEC: This strain is shipped as a test tube culture (10 ml of culture in a 20 x 125-mm screw-capped test tube). Upon arrival, rub the internal surfaces of the tube with a sterile cotton swab to dislodge any adhering amoebae. Aseptically divide the entire contents into 2 equal portions and distribute to T-25 tissue culture flasks. Add 5 ml of fresh ATCC medium 1405 and 0.5 ml of heat-killed Dunaliella tertiolecta ATCC 30929 to each flask. Screw the caps on tightly and incubate at 20-25C. Transfer every 4-6 weeks. Rub the surface of the flask to be subcultured with a sterile cotton swab, agitate and aseptically transfer a 0.25-0.5 ml aliquot to a fresh flask containing 10 ml of ATCC medium 1405 supplemented with 0.5 ml of heat-killed algal suspension. Incubate as above. Note: Since this strain is cultivated with heat-killed Dunaliella tertiolecta ATCC 30929 as a food source, the food should be prepared before the strain is ordered. The food is prepared as follows: 1. Establish a culture of ATCC 30929 in 5 ml of ATCC medium 1194 broth in a 16 x 125-mm screw-capped test tube. Incubate with the cap on loose under 50-75 (Einsteins/m2/s of light at 25C. 2. When the culture reaches early stationary phase, aseptically transfer 0.25-ml aliquots to each of 10 250-ml cotton-plugged Erlenmeyer flasks containing 100 ml of ATCC medium 1194 broth. Incubate as above. 3. When the cultures reach early stationary phase (approximately 10-14 days), harvest as follows: Aseptically transfer algal suspensions to 50-ml plastic centrifuge tubes. Centrifuge at 400-500 x g for 5 minutes. 4. Decant supernatant and resuspend each pellet in approximately one ml of ATCC medium 1405. Pool all suspensions in a single centrifuge tube and centrifuge as in step 3. The cell pellet should be approximately 5 ml. 5. Resuspend the cell pellet to a final volume of approximately 14 ml with ATCC medium 1405 and transfer to a 15-ml plastic centrifuge tube. Centrifuge as in step 3. Discard the supernatant. 6. Resuspend the cell pellet to a final volume of approximately 14 ml with ATCC medium 1405 and centrifuge as in step 3. Discard the supernatant. 7. Resuspend the final cell pellet to a final volume of 10 ml with ATCC medium 1405. Transfer the cell suspension to a sterile 125-ml screw-capped bottle and aseptically add 40 ml of ATCC medium 1405 bringing the final volume to 50 ml.WARN: Last ATCC#: 40318, unprocessed line: 8. Place bottle prepared in step 7 in a 60C water bath to a level such that the liquid level of the water bath is above that of the suspension in the bottle. Incubate for a total of 30 minutes, swirling the bottle at 10-minute intervals. Allow the bottle to cool to room temperature. This treatment will kill all algal cells. 9. As a check for viable cells, add 3 drops of the cell suspension prepared in step 10 to the edge of a 100-mm petri plate containing ATCC medium 1194 agar. Hold the plate vertically to allow the drops to move to the opposite edge. Incubate plate at 25C under 50-75 uE of light for 7 days. 10. The heat-killed algae can be stored at 4C for up to one year.
Name of Depositor M Polne-Fuller
References

Polne-Fuller M. Microoganisms and methods for degrading plant cell walls and complex hydrocarbons. US Patent 5,413,933 dated May 9 1995

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation