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Product Name:
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NK65
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Manufacturer:
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BEI Resources
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Taxonomy:
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Protozoa Classification: PIasmodiidae, Plasmodium
Species: Plasmodium berghei
Strain: NK65
Phenotype: Wild-type1
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Additional Information:
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P. berghei is a protozoan parasite that infects mammals other than humans, especially rodents, and is commonly used in rodent model studies of malaria. P. berghei preferentially invades reticulocytes, typically producing infections in mice that induce severe pathology.3,4
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Material Provided:
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Each vial contains approximately 0.5 mL of P. berghei-infected mouse blood in Glycerolyte 57 solution (1:2). This item is host-restricted and must be amplified in rodents. Please refer to Appendix I for cryopreservation instructions.
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Packing/Storage:
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MRA-268 was packaged aseptically in cryovials. The product is provided frozen and should be stored at -80°C or colder immediately upon arrival. For long-term storage, the vapor phase of a liquid nitrogen freezer is recommended (-130°C or colder). Freeze-thaw cycles should be avoided.
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Safety Precautions:
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All blood cultures should be handled with appropriate safety precautions necessary for the handling of bloodborne pathogens. Personnel must be trained in accordance with their institutional policy regarding bloodborne pathogens.
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Growth Conditions:
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In vivo, Swiss Webster mouse
Note: Some strains of mice may require dietary or drug pretreatment protocols for successful infection as P. berghei strains have a strong predilection for invasion of reticulocytes.
Inoculation5:
1. Thaw a frozen cryovial of MRA-268 in a 35°C to 37°C water bath for approximately 2 to 3 minutes. Do not allow the vial to immerse near the cap line seal while thawing.
2. Once thawed, wipe the outside of the vial with 70% ethanol before opening. Using a 1 mL syringe equipped with a 27-gauge 1/2-inch needle, remove approximately 200 µL to 300 µL from the vial.
3. Wipe the injection site of the mouse with 70% ethanol and inject the sample intraperitoneally at 50 µL to 100 µL per mouse (approximately 3 mice for most applications).
Monitoring parasitemia:
1. Starting 3 days post-inoculation, monitor the growth of parasites by tail vein bleed sampling and Giemsa-stained thin blood smear microscopy at 1- to 2-day intervals.
2. Passage the strain when the infection is at or near the first peak of parasitemia (> 5%). This will normally occur within one week of inoculation.
% parasitemia = (Infected RBC/Total RBC) × 100
Passaging:
1. Anesthetize infected mice by CO2/O2 inhalation. Collect the blood by orbital bleeding or from the tail vein into 25 mL of cold 1× PBS-heparin anticoagulant solution (please refer to Appendix I for preparation instructions).
2. Inject the sample into each of the uninfected mice (approximately 10 mice) as described in Inoculation step #3 above.
3. Monitor parasitemia as described above and passage as needed.
Please refer to Appendix I for cryopreservation instructions.
Note: Do not directly inject freshly thawed parasites from cryopreserved stocks by the intravenous (IV) route, as these samples contain cryoprotectant, anticoagulant and may contain traces of lysed or coagulated red blood cells. Direct IV inoculation from cryopreserved stock may result in pulmonary embolism or shock in mice.
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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. Nussenzweig, V., Personal Communication.
2. Ramiro, R. S., S. E. Reece and D. J. Obbard. “Molecular Evolution and Phylogenetics of Rodent Malaria Parasites.” BMC Evol. Biol. 12 (2012): 219. PubMed: 23151308.
3. Hall, N., et al. “A Comprehensive Survey of the Plasmodium Life Cycle by Genomic, Transcriptomic, and Proteomic Analyses.” Science 307 (2005): 82-86. PubMed: 15637271.
4. Otto, T. D., et al. “A Comprehensive Evaluation of Rodent Malaria Parasite Genomes and Gene Expression.” BMC Biol. 12 (2014): 86. PubMed: 25359557.
5. Peters, W. and B. L. Robinson (1999), “Chapter 92 -- Malaria.” In Handbook of Animal Models of Infection. Eds. O. Zak and M. Sande, Academic Press: London, pp. 757-773.
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Citation:
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Acknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Plasmodium berghei, Strain NK65, MRA-268, contributed by Victor Nussenzweig."
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Biosafety Level:
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1
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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