Clostridium ljungdahlii

平台编号：Bio-135382

Clostridium ljungdahlii Tanner et al. 菌种拉丁名称

BAA-3217™ ATCC编号

Clostridium ljungdahlii strain PETC is a bacterial type strain that was originally isolated from chicken yard waste. This product is an ATCC-manufactured and accessioned progeny of ATCC 55383 and ATCC 49587.

Product category 类别 Bacteria

Product type Anaerobe

Strain designation PETC

Type strainYes

Genome sequenced strain Yes

Isolation source Chicken yard waste

Geographical isolation United States; Arkansas

Product format 提供形式 Frozen

Storage conditions 保存温度 -80°C or colder 

 BSL 1 生物安全等级

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

Preceptrol No

Handling information

Medium 培养基 ATCC Medium 1754: PETC medium

ATCC Medium 2107: Modified Reinforced Clostridial

Temperature 培养温度 37°C

Atmosphere 需氧条件 Anaerobic 80% H2, 20% CO2

Handling procedure

Keep vial frozen until ready to use. This item will also grow on ATCC Medium #2107, Reinforced Clostridial Medium (BD 218081) broth and  ATCC Medium #1754, PETC broth.

To reduce media before inoculation, use 1.5% sodium sulfide M (0.2 mL per 10 mL).  Do not use 3% Cysteine as a reducing agent.

The 1.5% Sodium Sulfide solution (0.2 mL per 10 mL broth) needs to be added before inoculation. If needed, exchange the gas in the Hungate tube for 80% H2-20% CO2.  Tubes need to be pressurized to 20 psi. 

Under anaerobic conditions, preferably in an anaerobe chamber, thaw vial and then quickly transfer entire contents into a single Hungate tube of AS-822 broth. A second tube of AS-822 broth can also be inoculated with 0.5 mL from the original broth. 

Incubate at 35 to 37°C. Initial growth may be observed at 48 hours. Suitable growth for transfer may take up to 5 to 7 days.

Growth on agar plates is difficult because of the gas and pressure requirements. Better results can be achieved by producing agar slants in Hungate tubes. Agar cultures require a longer incubation period. 

Hungate tubes need to be re-gassed and pressurized after each transfer or manipulation of the culture. 

Growth is evident by light turbidity and an accumulation of cells at the bottom of the broth tube. A wet mount may be necessary to observe growth.  This strain needs to establish the growth first in broth for 4 days before transferring. This strain does not grow well on agar. No growth is observed when using TSA+5% SB agar or Brucella agar.

ANAEROBIC CONDITIONS:

Hungate (or Balch) tube refers to a special type of test tube that is designed to be pressurized and is suited for anaerobic work. The Hungate test tubes can be purchased form Bellco glass (www.bellcoglass.com; stock no. 204800150). 

Resazurin is a commonly used redox indicator that is pink when the redox potential is above -50 mv., and colorless when the redox potential is below -110 mv. i.e. highly reducing. Most strict anaerobes require this low redox potential for optimum growth. 

To obtain a fully reduced medium, it is necessary that the medium be anoxic and that a reducing agent be added. There are several common reducing agents, however, 1.5% sodium sulfide is recommended for this strain. Add 0.2 mL of reducing agent for each 10 mL of medium. 

Syringes can be made anaerobic by one of two methods. 

Displace the dead space in the syringe with a sterile oxygen-free gas.

Displace the dead space in the syringe with a reducing agent.

Handling notes

Tubes need to be pressurized to 20 psi.

Recommended to use AS-822 (PYG broth) from Anaerobe Systems.

This organism will also grow on ATCC Medium #2107, Reinforced Clostridial Medium (BD 218081) broth and ATCC Medium #1754, PETC broth.

Always use freshly prepared pre-reduced media or pre-reduced media that has been previously prepared but stored under anaerobic conditions. Sodium sulfate was found to be slightly inhibitory. Do not use 3% Cysteine as a reducing agent. Resazurin in the media is a color indicator for anaerobic conditions.  Observance of pink color in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred.  Medium should be discarded.

Additional information on this culture is available on the ATCC web site at www.atcc.org.

Quality control specifications

Verification method

Whole-genome Sequencing

History

Deposited as

ATCC accessioned progeny of Clostridium ljungdahlii strain PETC (ATCC 49587, ATCC 55383)

Depositors ATCC

Cross references

GenBank GU139552 Clostridium ljungdahlii strain ATCC 55383 16S ribosomal RNA gene, partial sequence

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.

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