Fighting the Biotech Wars

By Leon Jaroff.

In Farmer Kevin Main's farrowing barn near Altona, Ill., the newborn piglets lay on their sides, their tiny feet paddling frantically in the air. A day later they were dead. "It was not a pleasant thing," Main recalls. "We lost over a hundred." Main's 480-swine herd had been hit by pseudorabies, a disease caused by a herpes virus that attacks the central nervous system of pigs, sheep, cattle and other animals. Nearly always fatal in young pigs, it causes symptoms ranging from disoriented wandering to skin lesions to convulsions, and can lead to reproductive failure in animals that survive. About 10% of the nation's 54 million swine are currently infected with the virus, agriculture officials say, and the cost to the pork industry runs as high as $60 million a year.

Last week Main and other farmers were perplexed when the U.S. Department of Agriculture, under attack by critics, halted the sale of Omnivac-PRV, a new genetically engineered viral vaccine that immunizes swine against pseudorabies and may be the first of a whole generation of better animal vaccines. Main had participated in a field test of the vaccine last year by allowing 250 of his piglets to be inoculated. None of them or of others involved in the test contracted the disease, and in January the USDA licensed Biologics Corp., a vaccine manufacturer in Omaha, to put Omnivac on the market. "It seems to be safe," says Main. "I don't know what all the commotion is about."

The dustup was stirred by Jeremy Rifkin, a Washington-based gadfly and an implacable foe of genetic engineering (see box), who filed a petition with the USDA demanding suspension of Biologics' license. In issuing that license, Rifkin charged, the department had not only failed to follow federal guidelines for releasing live, genetically altered organisms into the environment but also neglected to conduct an environmental assessment of the risks involved. Stung by the petition and aware of Rifkin's uncanny success in obtaining court injunctions to back his demands, the USDA beat a hasty retreat. Bert Hawkins, administrator of the department's animal and plant health inspection service, conceded that the USDA did not "document" its environmental analysis because "we felt the product was not significantly different from products already on the market. In fact, it was a safer product. But we should have documented it for the public perception." That process, he said, should take only two weeks, after which the suspension would be lifted.

Rifkin objected. That the assessment could be done after licensing and in two weeks was "ridiculous," he said. "You can't now put together a hurried little paper and call it an environmental assessment. That's what you call cheating."

Rifkin's victory, however temporary, was still more bad news for the biotechnology industry, which has been stymied in its efforts to field test and market agricultural products containing genetically engineered live organisms. The defeats have been caused by a maze of confusing Government regulations, harassing legal actions and in some cases the industry's own blunders. Only last month, for example, the Environmental Protection Agency fined Advanced Genetic Sciences $20,000 and suspended its permit to field test a preparation called Frostban, which contains re-engineered bacteria designed to retard the formation of frost on plants. The agency charged that the Oakland firm misrepresented data and violated the national pesticide-control law by conducting outdoor tests without a permit. (EPA officials had been alerted by newspaper stories initiated by the indefatigable Rifkin.)

Overblown press stories and Rifkin's rhetoric about the two cases have raised the specter of re-engineered microbes escaping into the environment with dire consequences. But most scientists are convinced that neither the Biologics viral vaccine nor the A.G.S. bacteria pose any threat to man, beast or plant.

Like earlier vaccines against pseudorabies, Biologics' Omnivac consists of pseudorabies viruses altered to prevent them from causing disease but still capable of triggering production of the antibodies that make swine immune. While the viruses in other vaccines are rendered harmless by conventional methods, the Omnivac viruses are altered by recombinant-DNA techniques--in other words, by genetic engineering, or gene splicing. Saul Kit, the Baylor University biochemical virologist who redesigned the virus, points out that existing pseudorabies vaccines, which raise no alarms, have also been produced by what is really a form of genetic engineering. One older vaccine, he explains, was developed by growing the pseudorabies virus in chick cells for tens of thousands of generations. In the process, significant genetic changes occurred in small numbers of the virus. "The most important of these," he says, "was the deletion of an entire gene" that weakened the virus and made it suitable for a vaccine. What Kit has done by recombinant-DNA technology is to further disarm the virus by removing part of another gene that codes for an enzyme needed by the virus to reproduce.

Says Veterinarian Roger Saline, who supervised the test of Omnivac on Kevin Main's piglets: "I think they chose the wrong product to attack. The deletion work rendered the vaccine less virulent and less able to reproduce."

For A.G.S. executives, the suspension of their permit to field test Frostban marked still another in a series of frustrating delays. As long ago as 1982, the company began partly financing the research efforts of Steven Lindow and Nickolas Panopoulos, plant pathologists at the University of California, Berkeley, who were attempting to engineer bacteria that would inhibit the formation of frost on plants. Their long-range goals: to extend the growing season and reduce crop damage caused by unseasonal frosts, which costs U.S. farmers at least $1.5 billion every year.

The Berkeley team had focused its efforts on the bacterium Pseudomonas syringae, which lives on the leaves of many plants and actually promotes the formation of frost. As the temperature drops below 32 degrees F, specks of protein produced by the bacteria act as nuclei for the growth of ice crystals (see chart). Without the bacteria and their protein particles, plants can briefly sustain temperatures as low as 25 degrees F before the dew turns to frost. The solution seemed simple enough: from 2% to 5% of the bacteria in nature lack the ability to manufacture the protein. If large numbers of the microbes could be deprived of that capability, the frost threat could be eased.

Lindow and Panopoulos proceeded to expose large numbers of P. syringae to chemicals, and were able to impair in some of them the gene that orders production of the protein. When these altered microbes were sprayed on plants in greenhouses and open fields, they seemed to retard the formation of frost. Equally important, they apparently did not spread or do any harm, and most gradually died out. Their release into the open went unnoticed--or at least unchallenged--because they had been altered by conventional laboratory methods.

Still, the Berkeley team was not satisfied with its hit-and-miss method of creating frost-inhibiting bacteria, and set about producing them by gene- splicing techniques. For meaningful experiments, enough of the bacteria had to be sprayed on test plants to overwhelm the natural variety. But the release into the environment of any genetically engineered microbes in those days required the nod of the National Institutes of Health recombinant-DNA committee,* which in 1983 approved the Berkeley scientists' proposal to conduct their test at a tiny potato patch near the city of Tulelake in Northern California.

Enter Jeremy Rifkin, who with several allies filed suit in a federal court, charging that NIH, in granting its approval, had failed to prepare environmental-impact statements. At hearings conducted by Federal Judge John Sirica, Rifkin and a few scientists who supported his view pointed to the troubles caused in the U.S. by the introduction of foreign organisms

like the Japanese beetle and the gypsy moth. They warned that re-engineered P. syringae might also cause trouble. For example, they said, the altered bacteria might multiply and spread, perhaps even change the climate by retarding the formation of ice crystals in the atmosphere. Lindow and Panopoulos undermined that argument by pointing out that naturally occurring frost-inhibiting bacteria show no such inclination.

Still, Rifkin won the day. In May 1984 Judge Sirica issued an injunction against the Tulelake test on the ground that NIH had not followed the required procedure in granting its approval.

While that ruling was being appealed, A.G.S. was proceeding on its own. Late in 1984 it applied to the EPA for a permit to test Frostban on a strawberry patch in Monterey County, Calif.; last November the agency granted A.G.S. permission to spray some 8 trillion altered bacteria onto the field. But unknown to the EPA, the financially struggling company had already conducted outdoor tests of Frostban.

In the course of the safety testing, A.G.S. earlier this year had injected the frost-inhibiting bacteria into 48 fruit and nut trees. The company failed to report that the trees were not in a contained area but were growing in the open on the roof of its Oakland building. A.G.S. also neglected to tell the EPA that cankers (apparently caused by other organisms) had developed on some of the trees at the injection site. The EPA's subsequent penalties against A.G.S. were the first such actions it has taken against a biotechnology company. "We're sending a message," said Steven Schatzow, an EPA official. "This is a new technology that evokes great public concern. People who play this game had better be careful." Rifkin was elated. "It's a much tougher stand than we thought EPA would take."

Even before this gaffe, A.G.S. had been running into trouble at its proposed test site in Monterey County. The company had made little effort to educate the local citizenry about details of the test and refused to reveal the exact site. Fear fed on rumor, and in February officials imposed a 45-day ban on any spraying of Frostban; in March they renewed the ban. Said Sam Karas, chairman of the county board of supervisors: "I could not in good conscience allow A.G.S. to test in Monterey County."

Other biotech researchers and companies may benefit from the sad experience of A.G.S. Lindow and Panopoulos, now hoping to win approval for their potato field test at Tulelake by May, have given a series of lectures about their project before local farmers to allay any fears. Officials of Calgene, a biotechnology firm based in Davis, Calif., that plans to grow genetically engineered, herbicide-resistant tobacco plants, have talked frankly about its work to community officials and growers near the proposed test site.

A St. Louis-based industry giant, Monsanto, has taken special care to pave the way for a test it hopes to conduct as early as May at its research farm in nearby St. Charles County. Monsanto scientists have singled out a strain of the common soil bacterium Pseudomonas fluorescens that lives on the roots of corn plants. Using gene-splicing techniques, they have endowed it with a gene from another bacterium that produces a protein deadly to certain insects. Laboratory experiments have already shown that when seed corn is coated with the engineered P. fluorescens, the microbe colonizes the roots of the growing plants and with its toxic protein kills off the cutworms that feed on corn- plant roots.

After five years of development and 1,400 pages of data submitted to the EPA, the company is convinced that the doctored soil bacteria pose danger only to the cutworm. It has petitioned for a field test of the bacteria-coated seed corn and is telling its story to anyone who will listen. "We've got really concrete data that address legitimate concerns," says Monsanto's Robert Kaufman, director of crop protection and improvement. "We've tried to be open about it." The St. Charles County commissioners, state legislators and 150 area farmers have been treated to a tour of the proposed test plot, and local residents were invited to question Monsanto officials at a meeting in a nearby motor inn. Kaufman even gave a speech at a banquet sponsored by a farm booster club. "I feel like I'm almost a preacher," he says.

Monsanto officials, like others in the industry, are concerned that overregulation could stifle many of the young, struggling biotechnology companies and suggest that there is a limit to the patience of larger firms. "If we can't make these tests in a reasonable period of time," says Howard Schneiderman, Monsanto's senior vice president for research and development, "I'm going to give up and just not do it. If someone is going to worry about a tomato plant that will devour New York City or a microbial pesticide that will develop into plague, I can't justify spending millions of dollars a year on products I can't test."

FOOTNOTE: *Under current, overlapping regulations, frost-inhibiting bacteria are considered microbial pesticides and must be approved by the EPA for field testing or use. Gene-spliced animal biologics and plants require USDA clearance; human and animal drugs need Food and Drug Administration approval.

With reporting by Cristina Garcia/San Francisco and J. Madeleine Nash/Chicago