On Tuesday 8th August 2006, we received the equipment we needed in order to blood sample Koi, and centrifuge the samples so that they could be sent to CEFAS for ELISA testing. On the Wednesday and Thursday, we blood sampled each and every Koi in our unaffected ponds, which initially returned one positive result, and a few which were borderline. It turned out that the borderline results were due to the serum samples being red cell contaminated, and on retesting, these came back negative. The one positive that we had, however, stayed positive, so much so, that it was actually a higher reading than CEFAS's positive control. We put this Koi into a temporary pond we set up up, along with some 3 month old Tosai that we believed KHV naive, and temperature cycled it thoroughly three times, between 15c and 25c, and the Koi proved to not shed virus to the other Koi. Yet, back tracking this Kois history, it was exposed to the virus in Japan in Summer '05. We blood tested this Koi repeatedly up until Jan '07 to try to track what was heppening with it's anti-body level, which showed no sign of falling, even up the point when we sent it to CEFAS in Weymouth for research pusposes. CEFAS also tried to cause an outbreak with this Koi, to no avail. Yet, both CEFAS, and myself were confident that this Koi was a carrier, as it simply stands to reason, that if anti-bodies are still being produced, it has to be because the virus is present in the nervous system, and waiting to outbreak into the blood stream, where it would met by the abundance of anti-bodies. Theory has it, that a Koi such as this one is actually very safe, since it's immune system is very finely tuned, keeping the virus hidden. Whilst at CEFAS, they also ran blood tests to try to find traces of virus DNA, which turned up negative each time. Eventually, CEFAS took blood and tissue samples for long term storage, and euthanised the Koi.

Once we had blood sampled all of the Koi residing in our own ponds, we hit the road and went out testing all Koi that we felt may have been at risk in customers ponds, as well as many other Koi that we had sold over the previous 18 months, in order to try to establish where our problem came from, and to make certain that all Koi sold were safe. We hit an early snag, insofar as our original centrifuge had developed a fault, and refused to reach the required speeds for successful blood/sera separation. The problem with this machine meant that some Koi had to be sampled again once we had obtained a new machine. The machine pictured on the right is the old the faulty one. Our new machine is far superior to this one. During the course of this testing, we tracked down 4 Koi that resided in three clients ponds, that we deemed to be a risk. These Koi were subsequently euthanised. Once the clients Koi were all tested, we set about testing Koi that we had supplied to other dealers. The testing that we carried out revealed that the source of our KHV outbreak had in fact come from Tosai, as opposed to coming from some Nisai that we had first suspected as being to blame. Testing at two other dealers returned interesting results, as it concluded that one dealer had the same proportion of positives as the first, and in another case, a mild outbreak had been inadvertantly 'fixed'. With the kind co-operation of one of the dealers, we decided to see it it was possible to cause an outbreak with these Tosai. Strangely, an outbreak proved remarkably easy to induce, despite anti-viruside being present in the water! Please note, that back in May 2006, these Koi were properly temperature cycled, without inducing an outbreak! Scary stuff!

The testing we carried out Nationally, accrued almost £20,000 of expense, yet this was paled into inconsignificance compared to the value of the Koi we lost, and also those customers Koi we had to replace, as we handful here that we took to the Show to try to sell, and hence, lost those also on our return. What we were trying to do as a favour to a few clients, resulted in an additional £13,500 loss of Koi. It's not all bad news however, as our testing resulted in one client asking for his entire collection to be tested. This revealed two Koi that he had, that were very high positives, that weren't our concern. He then recommended three of his friends to have their Koi tested, and this revealed that all Koi that these four people had imported from a Japan trip in Nov '05, and collected in April '06, were all positive. It was a lucky escape for these people.