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Product Name:
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KETRI 2285
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Manufacturer:
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BEI Resources
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Taxonomy:
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Protozoa Classification: Trypanosomatidae, Trypanosoma
Species: Trypanosoma brucei subsp. rhodesiense
Strain: KETRI 2285
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Additional Information:
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T. brucei is a kinetoplastid protozoan parasite and is the causative agent of African trypanosomiasis, which is transmitted to both humans and livestock through the bite of the tsetse fly.3,4 The flies inject the infective stage (metacyclic trypomastigotes) from their salivary glands into the blood and lymphatic fluid of the host, where they undergo differentiation and enter the central nervous system by evading the host immune system through the use of antigenic variation of their surface glycoprotein coat.3,4
T. brucei is divided into three morphologically-identical subspecies that demonstrate distinct pathogenicities: T. brucei subsp. gambiense and T. brucei subsp. rhodesiense, which cause African sleeping sickness in humans, and the non-human infective T. brucei subsp. brucei.3,5 The production of vaccines against these diseases is difficult as a significant rise of resistance to trypanocidal drugs has been documented. Animal models, such as mice, are critical to understanding the mechanisms of the disease and also aid in the development of new therapeutic drugs.3
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Material Provided:
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Each vial of NR-46433 contains approximately 0.5 mL of trypanosome infected blood with 20% glycerol, 6.25% Yaeger’s anticoagulant and Trypanosome dilution buffer. Please see the Appendix for cryopreservation instructions and component details.
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Packing/Storage:
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NR-46433 was packaged aseptically in cryovials and is provided frozen on dry ice. The product should be stored at cryogenic temperature (-130°C or colder), preferably in the vapor phase of a liquid nitrogen freezer. If liquid nitrogen storage facilities are not available, frozen cryovials may be stored at -70°C or colder for approximately one week.
Note: Do not under any circumstances store vials at temperatures warmer than -70°C. Storage under these conditions will result in the death of the culture.
To ensure the highest level of viability, the culture should be initiated immediately upon receipt. Any warming of the product during shipping and transfer must be avoided, as this will adversely affect the viability of the product. For transfer between freezers and for shipping, the product may be placed on dry ice for brief periods, although use of a portable liquid nitrogen carrier is preferred. Please read the following recommendations prior to using this material.
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Growth Conditions:
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In vivo, BALB/c mouse
Inoculation:
1. Thaw a frozen ampule of NR-46433 in a 35°C to 37°C water bath for approximately 2 to 3 minutes.
2. Remove the contents of the ampule using a 1 mL syringe equipped with a 27 gauge 1/2 inch needle.
3. Inject 50 µL to 100 µL of the contents of the vial intraperitoneally into a mouse. Inject the remaining contents into a second mouse.
Monitoring parasitemia:
1. Bleed both mice at daily intervals to monitor parasitemia by microscopic examination using a haemocytometer and 0.85% ammonium chloride as diluent. Parasitemia may also be assessed by microscopic examination of blood films stained with a 5% Giemsa solution.
2. Passage the strain when the infection is at or near the first peak of parasitemia (≥ 1 × 108 parasites/mL or ≥ 30 parasites/high power field for Giemsa-stained blood films observed under 100×). This will normally occur after 2 to 4 days of inoculation. Notes: The rate of T. brucei subsp. rhodesiense infection may vary with the parasite strain and concentration of inoculum. Inoculation of mice with the KETRI 2285 strain often produces an acute and fatal parasitemia, which, depending on the size of the inoculum, is lethal within a week after injection.1
Passaging:
1. Anesthetize the first infected mouse by CO2/O2 inhalation. Collect the blood by orbital bleeding or from the tail vein using an anticoagulant such as Yaeger's anticoagulant solution (see Appendix) or EDTA.
2. Perform a parasite count and inject 5 × 104 to 1 × 105 parasites into each of the uninfected mice (approximately 10 mice).
3. Monitor parasitemia as described above and passage as needed.
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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. Bacchi, C. J., et al. “Differential Susceptibility to DL-α-Difluoromethylornithine in Clinical Isolates of Trypanosoma brucei rhodesiense.” Antimicrob. Agents Chemother. 34 (1990): 1183-1188. PubMed: 2118325.
2. Bacchi, C. J., Personal Communication.
3. Antoine-Moussiaux, N., S. Magez and D. Desmecht. “Contributions of Experimental Mouse Models to the Understanding of African Trypanosomiasis.” Trends Parasitol. 24 (2008): 411-418. PubMed: 18684669.
4. Peacock, L., et al. “Identification of the Meiotic Life Cycle Stage of Trypanosoma brucei in the Tsetse Fly.” Proc. Natl. Acad. Sci. USA 108 (2011): 3671-3676. PubMed: 21321215.
5. Turner, C. M., N. Aslam and C. Dye. “Replication, Differentiation, Growth and the Virulence of Trypanosoma brucei Infections.” Parasitology 111 (1995): 289-300. PubMed: 7567097.
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Citation:
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Acknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Trypanosoma brucei subsp. rhodesiense, Strain KETRI 2285, NR-46433."
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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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