Acanthoeca spectabilis

PRA-103 ^™

Product category: Protists
Classification: Opisthokonta, Choanomonada, Acanthoecidae
Strain designation: Ellis
Type strain: No
Isolation source: Salt marsh of Swan Bay; isolated April 18, 1996
Geographical isolation: Australia; Victoria; Queenscliffe
Product format: Frozen

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.

Characteristics

Comments: Phylogenetic analyses of SSU rRNA, LSU rRNA, tubA, and hsp-90

Handling information

Medium

ATCC Medium 1525: Seawater 802 medium

Instruction for complete medium

ATCC Media 1525 and 1405, combined in equal parts and inoculated with Klebsiella pneumoniae subsp. pneumoniae (ATCC^® 700831) or Enterobacter aerogenes (ATCC^® 13048)

Temperature

20°C

Culture system

Xenic

Incubation

With Enterobacter aerogenes ATCC 13048 as a food source.

Culture maintenance

Subculture every two weeks to a fresh T-25 flask of bacterized medium in the following manner:

1. Vigorously agitate the flask (or scrape the flask bottom using a sterile cell scraper) and aseptically transfer 0.5 ml from a growing culture to a T-25 tissue culture flask containing 10.0 ml of an equal-parts mixture of ATCC media 1525 and 1405 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC® 13048).

2. Incubate flask at 20-25°C with the cap on tightly.

Reagents for cryopreservation

RM-9 Media for cryopreservation of Tetrahymena

Proteose Peptone (Difco 0120) 5.0 g

Tryptone 5.0 g

K₂HPO₄0.2 g

Glucose 1.0 g

Liver extract 0.1 g

Glass distilled water 1.0 L

Dissolve components in glass distilled H₂O and autoclave.

Dryl?s Salt Solution

0.1 M NaH₂PO₄^.3H₂010.0 ml

0.1 M Na₂HPO₄^.7H₂010.0 ml

0.1 M Sodium citrate ^.2H₂0 15.0 ml

0.1 M CaCl₂ ^.2H₂0 15.0 ml

Distilled water 950.0 ml

Add the first 3 components to the distilled H₂O and mix thoroughly.

Add the CaC1₂ solution and mix thoroughly.

(Adding the solutions in the order indicated will avoid the precipitation of Ca salts.)

Cryopreservation

1. Transfer tetrahymena from usual growth medium to RM-9 medium and allow to grow to near peak density.

2. Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 800 x g for 5 min.

3. Adjust the concentration of cells at least 2 x 10⁶/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, do not leave in water bath, aseptically remove the contents of the ampule and inoculate into a T-25 tissue culture flask containing 10 ml of an equal-parts mixture of ATCC media 1525 and 1405 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC® 13048)..

9. Incubate at 20-25°C with the cap screwed on tightly.

10. Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 ml to 10.0 ml of fresh bacterized medium.

11. Follow the protocol for maintenance of culture.

History

Depositors: B Leadbeater
Type of isolate: Environmental
Year of origin: 1996

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

Leadbeater BS, et al. Choanoflagellate lorica construction and assembly: the nudiform condition. II. Acanthoeca spectabilis Ellis. Protist 159: 495-505, 2008. PubMed: 18485816

Leadbeater BS. Developmental and ultrastructural observations on two stalked marine choanoflagellates, Acanthoecopsis spiculifera Norris and Acanthoeca spectabilis Ellis. Proc. R. Soc. Lond. B Biol. Sci. 204: 57-66, 1979. PubMed: 37513

Carr M, et al. Molecular phylogeny of choanoflagellates, the sister group to Metazoa. Proc. Natl. Acad. Sci. USA. 105: 16641-16646, 2008. PubMed: 18922774

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