In a unique undercover operation worthy of a biotech-armed JamesBond, DNA sequencing has identified an unprecedented number ofmass-murder victims, and fingered their network of killers.The story began two years ago when Don White, founder and presidentof Earthtrust, approached marine population zoologist StephenPalumbi, and his post-doc, C. Scott Baker, at the University of Hawaii.Earthtrust is an international marine-wildlife conservation endeavor. Italready counted among its coups inciting a United Nations moratoriumon drift nets, which entangle and kill dolphins and porpoises asinnocent swimmers in commercial tuna fishing. White's spouse, SusieWhite, directs the movement's marine campaigns.Palumbi and Baker had spent several years using DNA genetic probesto start a reference collection of whales worldwide. "The Earthtrustpeople asked us," Palumbi told BioWorld Today, "whether ourtechnology could confirm their suspicions about illegal whale tradingaround the world, specifically in Japan. They basically came and said,`If you had a sample of whale meat, could you tell what species it was,and where in the world it came from?' We answered, `Sure. We can dothat.' "It took the pair of zoologists about a year to devise a practical protocol.Earthtrust would help them present their results to the 1994 meeting ofthe International Whaling Commission (IWC).Palumbi and Baker report the results of their project in the Sept. 9 issueof Science. Their paper is titled: "Which whales are hunted? Amolecular genetic approach to monitoring whaling."As Mrs. White recalled, "I telephoned John Hansen, director of specialprojects at M. J. Research Inc., in Watertown, Mass. and asked if theywould donate a polymerase chain reaction (PCR) machine. I could tellhim only that it was for a secret project."Hansen told BioWorld Today: "We took them on faith. They had donegood work exposing the slaughter of undersea life by drift-netters, so Ifigured they were doing something similar, and Scott vouched forthem."So the company donated a MicroCycler to Earthtrust. (The Sept. 16issue of Science will carry a full-page ad from M. J. Research, tellingthe story.)The Portable PCR CaperScott Baker took the PCR instrument in his luggage to Tokyo in thespring of 1993, and set it up in his hotel room. Earthtrust put him intouch with three trusted local cut-outs, who fanned out on a three-month retail shopping trip for whale-meat all over Japan's main island."Whale meat is fairly valuable stuff," Palumbi said. "Our agents paidupwards of $100 a pound for it on the market." He noted that Japan, amaritime nation, has a long history of whale-meat consumption, "forwhich they have great cultural affinity. Because of that, I guess thepoachers think it's worth taking risks to hunt illegally and smuggle in."Raw, cooked, smoked, shredded, marinated, in all its comestible forms,the operatives, familiar with the marketplace, found the legal andforbidden flesh on butcher's counters. When they didn't see it, theyasked for it, and it came out from under the counter.Many species of gourmet whale had reached near-extinction by 1982,when the IWC banned most commercial hunting, and set quotas on thecatch of others, limited to scientific research purposes only. ButPalumbi wryly observed that these restrictions were widely respectedonly in the breach. "The International Whaling Commission," he said,"is an organization held together by a thin veneer of consensus."The main loophole in its hunting quotas, he explained, through whichsmugglers drive their illegal catch, is the "scientific permit," whichenables countries to study whale population genetics and geographicdistribution."The IWC doesn't need to give a permit. It allows each country to giveitself a permit. Which gives you some idea of the agency's strength."License To KillIn Japan's case, "their excuse for killing and marketing the 300permitted minke -- smallest, fastest, most abundant -- whales a year isthat in the past, all their genetic research was done on alleles of liverallozymes, which they need to continue, to square with their previousdata. And for that, unfortunately, they point out, you have to kill theanimals to get a bit of its liver to analyze the allozymes."Terming this rationale "ironic and a little bit outrageous," Palumbiasserted: "There is absolutely no reason why one has to kill an animalto do liver allozymes. All our genetics of humpback whales has beenfrom skin biopsies. From free-ranging, completely unharmed animals,we get large amounts of DNA from the skin, and can look at virtuallyany gene we want to target."What's more, he emphasized: "Those 300 scientific-permit minkes areonly the ones the IWC knows about. Their legal sale on the Japanesemarket means that such a market is legal. Which in turn means there'spressure for smuggling and illegal hunting of poached meat, soldmixed with the legal permit product."Sequencing Genes Under CoverIn his hotel room, Scott Baker received 16 discreetly purchased meatsamples. In the donated PCR instrument he released enough DNA intosolution to process it.He amplified a portion of the mitochondrial DNA control region, usinga biotinylated primer, then bound the labeled cDNA to streptavidin-coated magnetic beads. He thoroughly washed all the whale's ownDNA, then he brought the beads back to Hawaii for analysis.Exporting whole whale meat or any of its products (including its DNA)across international boundaries is prohibited, but copied, synthesizedDNA is not. Earthtrust has kept all of its meat samples stashed inJapan, preserved in alcohol.Baker and Palumbi then sequenced the DNA on the beads, andcompared them with databases from their own work, and fromGenBank and European databases.Three of the 16 samples they assayed turned out not to be the baleenwhale species they expected. Two of them were dolphins, the third atoothed whale, all three edible.Their report in Science concluded that "products available currently onthe Japanese retail market may include species that have been importedillegally, and others [notably, humpbacks] that have been hunted orprocessed illegally. "An alternative interpretation of their data, they noted with veiled irony:That the humpback whale meat they found on sale had been in storagesince 1966, when IWC banned it.Baker, along with Earthtrust's Don and Susie White, presented thesefindings in May to the International Whaling Commission meeting inPuerto Vallarte, Mexico. Baker went as an unofficial member of theNew Zealand delegation."The scientists there," Mrs. White recalled, "were very interested in thetechnology Scott presented."The Netherlands delegation, supported by nine other countries,including the U.S., introduced a resolution based on the scientists'recommendations. It called on Japan "to restructure its researchprogram concerning (its 1994 scientific permit to catch) 100 minkewhales in the North Pacific in such a manner that the research interestsare adequately addressed with non-lethal methods, [i.e.] biopsysampling and DNA analysis."The resolution was formally adopted. However, Mrs. White said, theJapanese took exception and sent out their whale-killing ships anyway."For setting quotas in the Northern Hemisphere, the Baker/PalumbiDNA technology is ideal," she added. n

-- David N. Leff Science Editor

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