The Americans have come to the same conclusion...
Excuse me? The Americans have NOT come to the same conclusion. In fact, the Americans have NEVER tested any “BC salmon” for ISAv.
FYI… just for the record the State of Maine is required to test their salmon feedlots for ISAv and report their findings to the Feds. It also appears, since there hasn’t been any ISA and/or ISAv found in Maine since 2006 there is a good chance it has been eradicated. Want to know how it was done? We don NOT allow any salmon from NORWAY in the United States and… WE KICKED YOUR NORWEGIAN FRIENDS OUT OF OUR COUNTRY!
Now you don’t believe ISAv is in BC? How about explaining that to these people?
Transcript: ISA Day 1
Dr. Kibenge replied (starts on page 12, line 24):
“…in total we received… [four] submissions, the very first one was the 48 hearts of sockeye smolts. And in that testing we found two positive samples out of 48… the second set of samples was saved, we have from a different submitter and in that case I think we found in total three positive samples. And then in the third and the fourth, those samples were negative… So by the time we put a result, we are confident that is a true positive result.”
Dr. Nylund replied:
“among the first 48 I had one positive, and it was sample number 36…Among the others…There was one positive in the report from the 23rd, sample H10 and that was repeatable…And I also got sample 14 heart positive, but that was not possible to repeat… I had no sign of contamination…And it was only these tissues that came up positive. But of course I was not able to sequence any ISA virus from these samples. So I was not able to verify that this was actually ISA virus I was picking out. But you know that the assays that we are using…are very specific, so they should only be picking out ISA virus.”
Ms. Gagné replied: “They are negative.” Commission Counsel pursued this further, “At the bottom of the document there's a row… highlighted,…"Interpretation of DFO testing" is the heading, and then we see "inconclusive" or not applicable, depending. Were your RT-PCR results inconclusive?” Ms. Gagné replied,
“We reported them as inconclusive based on our policy. Samples are tested additionally for the quality of the RNA tissue, and in this case all samples submitted show extensive to total degradation of RNA.”
Brock Martland, then referred to an email from Ms. Gagné (page 17, line 7) that discussed a weak positive test found at the DFO Moncton lab. It included,
“I am not convinced it should be reported to our friends in Ottawa, guess why! We do not like to see a Ct like this, but this is the type of Ct that is equivalent to the finding by Nylund, i.e., can’t conclude anything from it.”
Dr. Miller elaborated that she conducted ISAV testing on wild salmon in the past using “an assay to segment 6, which does not necessarily pick up all strains of ISA,” and these results turned out negative. But after hearing of Dr. Kibenge’s positive ISAV results she was concerned that her lab “hadn’t done enough due diligence to make sure… [their] fish were negative.” Hence, Miller began retesting her sockeye samples after modifying her lab’s methods to include assays to segment 7 and 8 of the ISA virus. She reported,
“the ISA 7, P7 primer set amplifies the most positive samples...and we find quite a lot of variability in our ability to pick up positives with segment 8 with various segment 8 primers. But when we do pick them up, they sequence as being ISA. So I believe that what we have in B.C. is a somewhat divergent strain of ISA that is not universally picked up with all…the assays that are presently in use.”
Dr. Miller responded,
“…we provided a set of positive and negative blind samples on to Dr. Kyle Garver [DFO Pacific Biological Station]… he ran basically the same assay that Nellie Gagné [DFO Moncton] has run…and he also ran our ISA-7, the Plarre-7 primer sets that we use, and…he ran it under… their…protocol that is part of the validation protocol, and then also using the protocol that we use in our lab…And I should say that we have different instrumentation and a slight…variance in the protocol that we use for RT-PCR…[and] he was not able to pick up any positives using the DFO validated assay, but he did pick up a positive of ISA-7 using our assay with our pre-amplification.”
An interesting
briefing note by Dr. Miller discusses these details and other pertinent information.
Dr. Kibenge responded,
“…this work, at the time it was being carried out, there was no real time RT-PCR, so the testing that was done used the conventional RT-PCR, and the primers that were used were targeting segment 8 and they are the standard primers for testing for ISA… the results, I think, as far as I recall, were that Dr. Molly Kibenge was able [to get] several samples positive for ISA virus, and some of those samples were sequenced, the products that we amplified were sequenced, and deposited to the gene bank…So this was clearly a positive amplification of ISA virus in those samples.”
Dr. Kibenge replied,
“No, they were not published, and the reason that was given was that the results…we obtained were considered to have been due to contamination, and the decision to submit the paper was denied.”
Mr. Martland: “You say the results were considered to be attributed to contamination, considered that way by whom?”
Dr. Kibenge:
“Dr. Molly Kibenge was working in the lab of Dr. Simon Jones. Dr. Simon Jones was the supervisor of this project, and it was his call as to how to proceed with the results of that work.”
Mr. Martland then questioned Ms. Gagné about her involvement in the 2004 ISAV testing (page 26, line 40). She said the DFO Moncton lab tried to reproduce Molly Kibenge’s positive results but could not.