Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Isolation
clinical specimen - animal
Salvador Brazil
Isolation date: October, 1999
Product Format
frozen
Storage Conditions
liquid nitrogen vapor phase
Type Strain
no
Medium
RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate, 95%; horse serum, 5%
Growth Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Duration: The organism is propagated in Vero cells.
Protocol:
Scrape the Vero cells into the culture medium when holes develop in the monolayer (tachyzoite clusters are observed in the cells and 7 micrometer long banana-shaped tachyzoites are free in the medium).
Transfer approximately 0.5 ml of the harvested medium (or approximately 500,000 tachyzoites) to another T25 flask containing a fresh monolayer of vero cells.
Incubate at 37C.
Interval: approximately 1 week
Subcultivation
Protocol:
Scrape the Vero cells into the culture medium when holes develop in the monolayer (tachyzoite clusters are observed in the cells and 7 micrometer long banana-shaped tachyzoites are free in the medium).
Transfer approximately 0.5 ml of the harvested medium (or approximately 500,000 tachyzoites) to another T25 flask containing a fresh monolayer of vero cells.
Incubate at 37C.
Interval: approximately 1 week
Cryopreservation
Storage temperature: liquid nitrogen vapor phase
Name of Depositor
LF Gondim, MM McAllister
Special Collection
NSF - Protistology
Year of Origin
October, 1999
References
Barber JS, et al. Characterization of the first European isolate of Neospora caninum (Dubey, Carpenter, Speer, Topper and Uggla). Parasitology 111: 563-568, 1995. PubMed: 8559588
Gondim LF, et al. Isolation of Neospora caninum from the brain of a naturally infected dog, and production of encysted bradyzoites in gerbils. Vet. Parasitol. 101: 1-7, 2001. PubMed: 11587828
Gondim LF, et al. Variation of the internal transcribed spacer 1 sequence within individual strains and among different strains of Neospora caninum. J. Parasitol. 90: 119-122, 2004. PubMed: 15040677