Thanks Muhammad

   You mean at -20C

Thanks a lot Muhammad for sharing your valuable experience

Best regards

Swaid

Hi Anunraj

   You are doing your Masters from Vet para from GBPUAT, This world is so small, I have done my Masters from the same place.

Are you working under Dr. Stuti Vatsya, say my Greetings to her

 She is such a nice person and a great researcher, I have learned a lot from her

say my Hello to Dr. Rajiv Ranjan and Dr. Vidya Sagar

Cheers

Swaid

Freezing is the best solution (-80°C and transport on dry ice). Babesia is also quite resistant to cold. You can maintain Babesia-infected blood more than 1 week at 4°C and then culture them without too many problems if you have right conditions of culture (medium, red blood cells, serum...) 

I agree with Philippe, I have been able to culture protozoa that have been at 4 degrees Celsius for over a week.

Also when infected EDTA blood is shipped by regular mail, so not on dry ice, it has been feasible to start a culture upon arrival. (Shipment took a day or 4)

When I say that freezing is the best solution it is of course after addition of a cryopreservative solution. Shipping of infected whole blood at 4°C stays a very good alternative solution.

its best to keep it in liquid nitrogen aftercrypreservatio

It keep it alive for lpng term, we took the gas bottele

Survival of Babesia in vitro - bound to be different for each species of Babesia;

 - Temperature (lower the better but d0 not freeze unprocessed infective blood);

 - Number of parasites in the sample (obtain maximum possible by collecting      blood from animal with clinical babesiosis when parasites are easily detected in blood films;

 - Use splenectomised donors if possible.

We found infected blood containing large numbers of B.bovis at collection was still infective after several weeks storage at about 50 C.

If infected blood is not chilled, glucose is used up and parasites die quickly.

(see GEFarlow 1976, Int Jour for Parasitol 6: 513-516. RJDalgliesh is the authority on the freeze preservation of Babesia using cryoprotectants.

For in vitro cultivation of Babesia see P Timms & NP Stewart 1989, Research in Veterinary Science 47: 309-310.

Good luck

Bill Callow

Alsever s solution for about 5 days in 4 C

After collection in evacuated tubes with anticoagulants, they tend to dissappear in few hours. But we have preserved them alive for weeks al 4°C, when the infected erythrocytes are suspended in  VYM´s solution. Please see: Vega et al. 1985 Am J Vet Res 46:421-423. 

Very much grateful for your help. I am planing to maintain a babesia

I agree with Sadegh. You can collect infeted bool in tubes containing sterile alserver's solution, which works as anticoagulant, but keep them at 4° C.

A pleasure to exchange communications.  

EDTA is fine as well

After collecting blood you can store at 4 C with anticoagulant. Storage procedure used for other blood intracellular protozoa can be applied here. 

dear researchers,

i am glad to inform you that we are getting positive samples of babesiosis from field cases and we are trying to propagate them. thank you for your valuable support. is there any thing else that i should be careful while culturing