Protostelium mycophaga Olive et Stoianovitch

PRA-154 ^™

Product category: Protists
Classification: Amoebozoa, Eumycetozoa, Protostelia
Strain designation: Type
Type strain: No
Isolation source: Dead inflorescence of common reed, Phragmites australis (Cav.) Steudel, in meadowland
Geographical isolation: United States; New Jersey

Documentation

BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

General

Specific applications: Phylogenetic analysis

Characteristics

Comments: Phylogenetic analysis

Handling information

Medium

ATCC Medium 2432: wMY (weak Malt Yeast Extract)

Instruction for complete medium

ATCC Medium 2432 inoculated with Rhodotorula mucilaginosa (ATCC^® MYA-3510)

Temperature

15-20°C

Incubation

Grown with Rhodotorula mucilaginosa ATCC MYA-3510 as food source

Handling procedure

Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen. If liquid nitrogen storage facilities are not available, frozen ampules may be stored at or below -70°C for approximately one week. Do not under any circumstance store frozen ampules at refrigerator freezer temperatures (generally -20°C). Storage of frozen material at this temperature will result in the death of the culture.

1. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes. Do not agitate the ampule. Do not leave ampule in water bath after it is thawed.

2. Immediately after thawing, aseptically transfer contents to a plate of ATCC medium 2432. Distribute the material evenly over the plate using a spread bar.

3. Wrap the entire edge of the plate with parafilm and incubate upright at 25°C. Amoeboid trophozoites and/or stalked fruiting bodies should be seen within 2-5 d.

Culture maintenance

1. Prepare an ATCC medium 2432 plate with a lawn of Rhodotorula mucilaginosa (ATCC^® MYA-3510) and incubate at 25°C overnight.

2. Remove an agar block (~5 mm² ) with trophozoites from the edge of an agar plate culture and invert the block at the edge of the freshly prepared plate.

3. Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.

4. Repeat steps 1-3 at 7-10 d intervals.

Reagents for cryopreservation

Cryoprotective Solution

DMSO 1.5 ml

Fresh growth medium w/o bacteria 8.5 ml

Cryopreservation

1. Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.

2. Harvest cells from a culture which is at or near peak density by adding 5 ml ATCC medium 5080 (Dryl's solution) and washing cells into suspension. Rub the surface of the plate with a spread bar to detach adhering trophozoites.

3. Adjust the concentration of cells to at least 2 x 10⁴/ml in fresh medium.

4. Mix the cell preparation and the cryoprotective solution in equal portions.

5. Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

6. Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1 C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.)

7. Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8. To establish a culture from the frozen state place the vial in a 35°C water bath. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial. Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 2432. Distribute the material evenly over the plate using a spread bar.

9. Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.

10. Follow the protocol for maintenance of culture.

History

Deposited as: Protostelium mycophaga Olive et Stoianovitch
Depositors: FW Spiegel
Type of isolate: Plant
Year of origin: 1959
Special collection: Eumycetozoan Project

Legal disclaimers

Intended use

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

Warranty

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Phragmites australis (Cav.) Steudel (Phragmites communis, common reed) dead inflorescence, New Jersey meadowland

Olive LS. The Protostelida - a new order of the Mycetoozoa. Mycologia 59: 1-29, 1967.

Olive LS, Stoianovitch C. Two New Members of the Acrasiales. Bull. Torrey Bot. Club 87: 1-20, 1960.

Shadwick LL, et al. Eumycetozoa=Amoebozoa?: SSUrDNA phylogeny of protosteloid slime molds and its significance for the amoebozoan supergroup. PLoS One 4(8): e6754, 2009.

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