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Product Name:
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Bosnia A (in vitro)
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Manufacturer:
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Steven J. Norris, Ph.D., Professor and Vice Chair for Research, Department of Pathology and Laboratory Medicine, University of Texas Health Science Center at Houston McGovern Medical School, Houston, Texas, USA, supported under NIH Grant R01 AI141958
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Taxonomy:
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Bacteria Classification: Treponemataceae, Treponema
Species: Treponema pallidum subsp. endemicum
Strain: Bosnia A
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Additional Information:
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T. pallidum is a pathogenic, obligate spirochete and the causative agent of syphilis (T. pallidum subsp. pallidum), bejel or endemic syphilis (T. pallidum subsp. endemicum) and yaws (T. pallidum subsp. pertenue) in humans.3,4,5 Long-term propagation of T. pallidum has been limited to rabbits since the bacterium was first identified in 1905.7,8,9 The recent optimization of an in vitro method in Sylvilagus floridanus epithelial cells with a specialized growth medium has allowed for continuous propagation while maintaining the characteristic ultrastructure and full infectivity in vivo.5,9
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Material Provided:
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Each vial of NR-60824 contains approximately 1.25 mL of bacteria extracted from infected rabbit tissue provided in TpCM-2 medium containing 20% heat-inactivated fetal bovine serum and 10% glycerol. The concentration is reported on the Certificate of Analysis.
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Packing/Storage:
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NR-60824 was packaged aseptically in cryovials. The product is provided frozen and should be stored at -60°C or colder immediately upon arrival. For long-term storage, the vapor phase of a liquid nitrogen freezer is recommended. Freeze thaw cycles should be avoided.
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Growth Conditions:
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Media:
T. pallidum Culture Medium 2 (TpCM-2)5 (Appendix I)
Host:
Sylvilagus floridanus (cottontail rabbit) epithelial cells (Sf1Ep; ATCC® CCL-68™)
Incubation:
Temperature: 34°C
Atmosphere: Microaerophilic (1.5% O2; 5% CO2; 93.5% N2)
Please refer to the Appendix document containing Appendix II for preparation of host cell monolayer, Appendix III for propagation of T. pallidum, Appendix IV for maintenance of T. pallidum cultures and Appendix V for cryopreservation instructions.
Note: Complete protocols and additional information and instructions are provided in: Edmondson, D. G. and S. J. Norris. “In Vitro Cultivation of the Syphilis Spirochete Treponema pallidum.” Curr. Protoc. 1 (2021): e44. PubMed: 33599121. Please refer to this reference before starting work with NR-60824.
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Disclaimers:
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You are authorized to use this product for research use only. It is not intended for human use. Use of this product is subject to the terms and conditions of the BEI Resources Material Transfer Agreement (MTA). The MTA is available on our Web site at www.beiresources.org. While BEI Resources uses reasonable efforts to include accurate and up-to-date information on this product sheet, neither ATCC® nor the U.S. Government makes any warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. Neither ATCC® nor the U.S. Government warrants that such information has been confirmed to be accurate. This product is sent with the condition that you are responsible for its safe storage, handling, use and disposal. ATCC® and the U.S. Government are not liable for any damages or injuries arising from receipt and/ or use of this product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, the U.S. Government, ATCC®, their suppliers and contributors to BEI Resources are not liable for damages arising from the misidentification or misrepresentation of products. |
References:
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1. Norris, S. J., Personal Communication.
2. Staudová, B., et al. “Whole Genome Sequence of the Treponema pallidum subsp. endemicum Strain Bosnia A: The Genome is Related to Yaws Treponemes but Contains Few Loci Similar to Syphilis Treponemes.” PLoS Negl. Trop. Dis. 8 (2014): e3261. PubMed: 25375929.
3. Edmondson, D. G., et al. “Clonal Isolates of Treponema pallidum subsp. pallidum Nichols Provide Evidence for the Occurrence of Microevolution During Experimental Rabbit Infection and In Vitro Culture.” PLoS One 18 (2023): e0281187. PubMed: 36917571.
4. Edmondson, D. G., et al. “Parameters Affecting Continuous In Vitro Culture of Treponema pallidum Strains.” mBio 12 (2021): e03536-20. PubMed: 33622721.
5. Edmondson, D. G. and S. J. Norris. “In Vitro Cultivation of the Syphilis Spirochete Treponema pallidum.” Curr. Protoc. 1 (2021): e44. PubMed: 33599121.
6. Lukehart, S. A. and C. M. Marra. “Isolation and Laboratory Maintenance of Treponema pallidum.” Curr. Protoc. Microbiol. 12 (2007): 12A.1. PubMed: 18770607.
7. Nichols, H. J. “Observations on a Strain of Spirochaeta pallida Isolated from the Nervous System.” J. Exp. Med. 19 (1914): 362-371. PubMed: 19867775.
8. Nichols, H. J. and W. H. Hough. “Demonstration of Spirochaeta pallida in the Cerebrospinal Fluid From a Patient with Nervous Relapse Following the Use of Salvarsan.” JAMA 60 (1913): 108-110.
9. Edmondson, D. G., B. Hu and S. J. Norris. “Long-Term In Vitro Culture of the Syphilis Spirochete Treponema pallidum subsp. pallidum.” mBio 9 (2018): e01153-18. PubMed: 29946052.
10. Norris, S. J. and D. G. Edmondson. “Compositions and Methods For Long-Term In Vitro Culture of the Syphilis Spirochete.” U.S. Patent No. 11,130,937 B2, 2021.
11. Turner, T. B. and D. H. Hollander. “Biology of the Treponematoses Based on Studies Carried Out at the International Treponematosis Laboratory Center of the Johns Hopkins University under the Auspices of the World Health Organization.” Monogr. Ser. World Health Organ. (1957): 3-266. PubMed: 13423342.
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Citation:
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Acknowledgment for publications should read “The following reagent was obtained through BEI Resources, NIAID, NIH: Treponema pallidum subsp. endemicum, Strain Bosnia A (in vitro), NR-60824.”
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Biosafety Level:
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2
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories (BMBL). Current Edition. Washington, DC: U.S. Government Printing Office.
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