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Product Name:
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WA1
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Ownership statement:
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This reagent is the tangible property of the U.S. Government.
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Manufacturer:
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BEI Resources
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Taxonomy:
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Protozoa Classification: Babesiidae, Babesia
Species: Babesia duncani (also referred to as WA1-type Babesia1)
Strain: WA1
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Additional Information:
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Babesia species are intraerythrocytic protozoan parasites of the phylum Apicomplexa that are the causal agents of babesiosis, which is transmitted to both humans and mammals by infected ixodid ticks.4,5 Infection with Babesia species is usually asymptomatic or can result in mild flu-like symptoms that subside within a few days. Severe cases featuring acute anemia, thrombocytopenia, organ failure, or even death may occur among the elderly, splenectomized and immunocompromised individuals.4,5 The majority of human cases of babesiosis in the United States are caused by B. microti, while B. divergens is the primary cause of babesiosis in Europe, though human infections caused by B. divergens-like parasites in the United States have been reported.5,6,7,8 B. duncani infections in the United States have occurred through both tickborne and blood transfusion routes.9
In mouse and hamster models of infection, B. duncani causes acute disease with a sudden and severe increase in parasitemia resulting in death.10
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Material Provided:
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Each vial of NR-12311 contains approximately 0.5 mL of B. duncani-infected hamster blood in Alsever’s solution containing 10% glycerol. Please refer to the Appendix for cryopreservation instructions and component details.
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Packing/Storage:
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NR-12311 was packaged aseptically in cryovials and is provided frozen on dry ice. The product should be stored at -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, Golden Syrian hamster
Inoculation:
1. On the day of inoculation, thaw a frozen vial in a 35°C to 37°C water bath for approximately 2 to 3 minutes.
2. Remove the contents of the vial using a 1 mL syringe equipped with a needle.
3. Inject the contents of the vial into one mouse by facility-approved methods.
Assessment of infection:
1. Monitor the infection at 2 to 3-day intervals by microscopic examination of blood films stained with Giemsa solution.
2. Count the number of infected red blood cells (RBC) versus the total number of red blood cells under oil immersion and determine the % parasitemia:
% parasitemia = (Infected RBC/Total RBC) × 100
Note: A minimum of 500 red blood cells should be counted.
3. Passage the strain when the level of parasitemia is ≥ 10%. This will normally occur 1 to 2 weeks post-inoculation, but the rate of infection may vary.
Note: The rate of infection and level of parasitemia before the host succumbs is dependent on the strain and concentration of inoculum. Regular monitoring is recommended.
Passaging:
1. Anesthetize the first infected mouse and collect blood by facility-approved methods. Collect the blood using an anticoagulant such as Yaeger's anticoagulant solution (Appendix) or EDTA.
2. Dilute the blood 1:5 or 1:10 in Alsever's solution (Appendix: Babesia In Vivo Cryopreservation) and inject 0.1 mL to 0.2 mL of the infected blood suspension into each new mouse.
3. Monitor parasitemia as described above and passage as needed.
Please see the Appendix for cryopreservation instructions.
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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. Conrad, P. A., et al. “Description of Babesia duncani n. sp. (Apicomplexa: Babesiidae) from Humans and Its Differentiation from Other Piroplasms.” Int. J. Parasitol. 36 (2006): 779-789. PubMed: 16725142.
2. Quick, R. E., et al. “Babesiosis in Washington State: A New Species of Babesia?” Ann. Intern. Med. 119 (1993): 284-290. PubMed: 8328736.
3. Thomford, J. W., et al. “Cultivation and Phylogenetic Characterization of a Newly Recognized Human Pathogenic Protozoan.” J. Infect. Dis. 169 (1994): 1050-1056. PubMed: 8169390.
4. Leiby, D. A. “Transfusion-Transmitted Babesia spp.: Bull's-Eye on Babesia microti.” Clin. Microbiol. Rev. 24 (2011): 14-28. PubMed: 21233506.
5. Vannier, E. and P. J. Krause. “Human Babesiosis.” N. Engl. J. Med. 366 (2012): 2397-2407. PubMed: 22716978.
6. Beattie, J. F., M. L. Michelson and P. J. Holman. “Acute Babesiosis Caused by Babesia divergens in a Resident of Kentucky.” N. Engl. J. Med. 347 (2002): 697-698. PubMed: 12200568.
7. Herwaldt, B. L., et al. “A Fatal Case of Babesiosis in Missouri: Identification of Another Piroplasm that Infects Humans.“ Ann. Intern. Med. 124 (1996): 643-650. PubMed: 8607592.
8. Herwaldt, B. L., et al. “Babesia divergens-Like Infection, Washington State.” Emerg. Infect. Dis. 10 (2004): 622-629. PubMed: 15200851.
9. Herwaldt, B. L., et al. “Transfusion-Associated Babesiosis in the United States: A Description of Cases.” Ann. Intern. Med. 155 (2011): 509-519. PubMed: 21893613.
10. Abraham, A., et al. “Establishment of a Continuous in vitro Culture of Babesia duncani in Human Erythrocytes Reveals Unusually High Tolerance to Recommended Therapies.” J. Biol. Chem. 293 (2018): 19974-19981. PubMed: 30463941.
11. Schuster, F. L. “Cultivation of Babesia and Babesia-Like Blood Parasites: Agents of an Emerging Zoonotic Disease.” Clin. Microbiol. Rev. 15 (2002): 365-373. PubMed: 12097245.
12. Uilenberg, G. “Babesia – A Historical Overview.” Vet. Parasitol. 138 (2006): 3-10. PubMed: 16513280.
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Citation:
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Acknowledgment for publications should read “The following reagent was obtained through BEI Resources, NIAID, NIH: Babesia duncani, Strain WA1, NR-12311.”
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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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