We currently get our blood supplies from a veterinary supply company but it only lasts a couple of weeks and we've have had some rather nerve-wracking supply interruptions in the past. Can anyone out there recommend any methods of preserving blood that can then be used to feed insect (or tick) colonies? (We tried simply freezing it and thawing it, with fairly poor results)
I've never heard of freezing blood for subsequent feeding, though I work on mosquitoes and not ticks. Anyway, simply freezing and thawing blood will induce RBC lysis, so you might want to play around with cryopreservation agents (glycerol, DMSO...). You might need to think of changing your supplier to get more consistent supplies?
Thanks Jean! Yes, I realised we would get RBC lysis after freeze-thaw, but my primary interest is Stomoxys calcitrans maintenance - these are very unfussy (you can keep them on isotonic sports drinks for a while if blood is short) but consume more blood than all our mosquitoes, Culicoides and ticks combined, so I thought it was worth a try. I'd heard that glycerol could work but wondered if anyone had a protocol; if not we can play around and try to come up with one.
Sadly our supply problems have been partly caused by our e-purchasing system so we can't do much about this, but it would also save money to buy in bulk. Some of our mosquitoes also need human blood, and the supply of expired blood from blood banks is unpredictable by nature, so we're quite keen to look into this.
We do cryopreservation using glycerol to preserve Plasmodium parasites grown in culture in RBC. You could try to adapt it to your blood. The cryopreservation solution is made of 3M glycerol, 0.16M sorbitol, 0.11M NaCl. We mix our RBC pellet (5min, 500g) with an equal volume of this solution, dropwise, very slowly, constantly under agitation, then aliquot in cryovials and snap-freeze in liquid Nitrogen and store in N2. Hope this helps and let me know if you have successful feeding after that.
Hi,
Have you tried CPDA method for whole blood preservation. You could probably store the whole blood (with minimal effect on RBC lysis) for two months. You might purchase larger volumes of blood at once (depends on your requirement; delay time in your next procurement) which could suffice until your next order and then store whole blood using the CPDA method. But, I believe you need to check with supplier where they have put any other anticoagulant. I run a mosquito insectary where we bring fresh blood from veterinary by adding CPDA into the collective vessel before we draw blood.
Alternatively, you could ask your supplier to add CPDA to fresh blood collection (you might be interested to consider dispatching CPDA solution prior to the anticipated date of blood collection).
Hope this will help you!
I cannot recall the supplier we used while I worked at Keele in Paul Eggleston's group (on An gambiae) but I can find out if you are interested. In the US, we have had very reliable supply of defribrinated sheep's blood for our Aedes colonies We are currently trying feeding frozen-thawed blood to our Aedes aegypti as we heard someone was successful (but then they are not fussy at all when compared to other species).
We use defibrinated horse blood ( certified pathogen free) from TCS Biosciences for all our mosquitoes ( Aedine and Anopholine) here in the UK - 1 litre bags are available
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