Bacteria Tests Await Chance to See Daylight

Last spring a young woman unwittingly released 30 trillion unidentified foreign bacteria in the yard of her new home. Steve Lindow calls her act “far more dangerous” than what he wants to do.

Lindow is a scientist at the University of California, Berkeley, who has been trying since 1983 to get legal permission to spray gene-spliced bacteria on potato plants in a field in California.

The young woman’s illegal act was transplanting wildflowers from Michigan to Virginia in a bucket full of soil. Lindow said the bucket of soil would have contained about the same number of live bacteria as he plans to release, “but we’re only using native California bacteria, and we can characterize the bacteria present. There’s far less hazard.”

If Lindow’s work is less dangerous, it also has a more serious purpose than transplanting posies. He wants to test the bacteria’s power to reduce frost damage to plants and, at the same time, test the regulatory system that will govern the release of genetically engineered creatures into the environment.

If the anti-frost bacteria pass their field test, some estimates say they could save farmers $1 billion a year in crops now lost to early freezing.

So far, all that has been tested is the plant pathologist’s patience and the regulatory system. The bacteria will remain in the lab at least until next year.

In a court suit aimed at blocking release of the organisms, the university agreed Aug. 20 to temporarily drop plans to test them and to meet with organizations and individuals who brought the suit.

University officials said it was too late in the season to begin the experiment.

Other scientists trying to get approval for a first release of genetically engineered bacteria have faced similar lengthy encounters with federal agencies, including the National Institutes of Health, the Environmental Protection Agency, the U.S. Department of Agriculture, and state and federal courts.

One company, Advanced Genetic Systems Inc. of Oakland, received a boost recently in its effort to use frost-proof bacteria on strawberry plants. The EPA said the company’s indoor test results indicate “no foreseeable adverse impacts to human health or the environment are likely to result.”

But the EPA also said the company must first select a site for its outdoor strawberry testing before the federal agency will consider issuing a test permit.

The company had hoped to run the test on a strawberry field in Monterey County, but local officials protested and the experiment was canceled. Advanced Genetic Systems said a new site has not been selected.

Jeremy Rifkin, the most conspicuous and vocal opponent of releasing organisms with “designer genes,” predicted that Advanced Genetic Systems will run into more citizen resistance for its strawberry testing “because no one wants to be the first guinea pig.”

It was his group, the Washington-based Foundation on Economic Trends, that filed suit against Lindow’s experiment.

For the frost resistance, scientists use the natural plant bacterium Pseudomonas syringae and remove a gene that makes a protein that acts as a nucleus for ice-crystal formation. Scientists hope that the altered bacteria will colonize on the plants and crowd out the bacteria containing the frost-triggering protein.

The legal battles may have been exasperating to the genetic engineers, but they have clarified the regulations and provided a forum for discussing potential hazards of releasing genetically engineered organisms.

The debates have pitted non-scientists, who fear that the gene-splicing technology may have unforeseen risks, against scientists who, like Lindow, see virtually no risk in his carefully regulated research.

That there even is an issue shows how far biotechnology has come in a decade.

Scientists use genetic engineering techniques to make direct changes in the DNA blueprint of cells. After they have identified and characterized a gene that gives a cell a specific trait, they use chemical scissors to manipulate the gene. Genes can be snipped out, moved to another cell and chemically knitted into the genetic plan of a different variety or a different species.

With such techniques, researchers can produce in a few months what could take years to obtain through traditional genetic techniques.

The older methods rely on rare natural or induced mutations to produce a desired trait and then may require generations of breeding to combine the gene with other desirable genes.

Genetic engineers may also move genes across breeding boundaries that nature and traditional genetic techniques could never cross. Human growth hormone genes, for example, have been inserted into the genetic blueprint of mice, and disease-resistance genes from bacteria have been inserted into tobacco plants.

On June 26, President Reagan’s Office of Science and Technology Policy released a new framework of regulations governing the new technology and spelling out which agencies have authority over each type of genetic engineering project. The new framework also defines which organisms will come under the strictest scrutiny.

Hearings on the new rules and a continuing series of lawsuits filed by opponents of genetic engineering have forced scientists to consider--and speculate on--the environmental risks of releasing genetically engineered organisms.

“We’ve developed regulations for an industry that has never shown a need for regulation,” said David Kingsbury, who headed the White House panel that developed the new framework of rules. “We’re regulating imagined disasters rather than real ones and have no reason to think there will be a problem.”

Rifkin does anticipate problems.

“What we’re trying to do is raise the environmental, economic, social and ethical questions about an entire technology revolution in advance of its coming on line,” he said.

In this never-never land of speculation, there are few answers that are absolutely right or totally implausible, and personal politics often dictate how people predict the future.

“Some of the outspoken critics work hard to find problems with new applications of biotechnology,” said Mike Botchan, a molecular biologist at Berkeley.

“People who use the technology and have a vested interest in the success of a trial will pooh-pooh predicted problems,” he said. “It’s a question of whether you’re an optimist or a pessimist.”

Botchan, who uses gene-splicing techniques in his work, is an optimist: “I have faith in the EPA and disinterested federal employees. If they say it’s safe, they’ve probably addressed the issues adequately.”

Liebe Cavalieri, a molecular biologist at Sloan Kettering-Memorial Cancer Center in New York, is more pessimistic and has decided not to use recombinant DNA technology in his laboratory studies.

“The regulations leave a lot to be desired,” he said.

Cavalieri, although worried about the new regulations and Lindow’s proposed study, does not oppose the eventual release of genetically engineered bacteria.

“When you talk about ecological questions, you have to do it in the field,” he said. “We shouldn’t make these studies against the law, but we should force them to set up experiments that are sensible.”

Other scientists, including ecologists, share Cavalieri’s refusal to oppose releases categorically.

University of Minnesota ecology professor Phil Regal said, “We cannot stop genetic engineering, but we must learn to deal with the risk. There are lessons to be learned.”

Chief among these lessons, Regal said, are basic facts about bacteria and their ecology.

“We are gambling,” Regal said. “We know remarkably little about bacteria.

“If you took a bit of soil and washed out the bacteria, you might find 30 different types. Of these, scientists would only be able to identify and name two or three and might not be able to grow any of them in culture.”

Growing bacteria in culture is one of the first steps necessary to studying them.

The government’s National Science Foundation has tacitly admitted the inadequacies of microbiology outside the lab and “made a pile of money available to develop microbial ecology” as a field of study, according to Richard Root, president of the Ecological Society of America.

But Root does not see this investment paying off immediately.

“NSF is trying to encourage people in the area, to make it attractive to go into it, but there is a time lag,” Root said. He does not expect to see the real flowering of microbial ecology for at least five years.

In the meantime, Regal has been working “to get the scientific infrastructure in place” for microbial ecology and has tried to foster communication between microbiologists and ecologists. He hopes the two groups of scientists will be able to arrive at a consensus on the best techniques for studying microbes in nature and how to make predictions about their growth.

Given the dearth of information about how genetically altered bacteria will behave out in the field, many ecologists base their guesses on analogies with unaltered organisms that have been deliberately or accidentally introduced from abroad.

Elliot Norse, spokesman for the Ecological Society of America, said there are two potential hazards with new organisms in the environment: They might prove to be too competitive and successful or they might pass new genes on to other species.

“In the majority of cases there’s not going to be a problem,” Norse said. “Based on experience with introduced plants and animals, we know that most introductions fail. In a small number of cases, they don’t fail, and in a fraction of these cases there’s a real problem (with the organism spreading out of control).”

Norse cited the example of the kudzu vine and the gypsy moth, which were introduced in the United States to control soil erosion and make silk. Both have become serious pests.

John Wood, director of the biotechnology and environmental coordination staff of the Animal and Plant Health Inspection Service said, however, that since the adoption of the Plant Pest Act in the 1950s, there have been no catastrophic introductions, although thousands of new organisms have been approved for import into the United States.

Wood said the potential spread and aggressiveness of a species are always considered before the U.S. Department of Agriculture will allow an organism to be released.

Under the new regulations, genetically engineered plants would have to undergo an evaluation similar to that for foreign plants before they could be grown outdoors.

But scientists can only speculate on the gene-transfer question. Opponents of environmental release of altered organisms point to the transfer of drug-resistance genes in bacteria.

They fear that a microbe engineered to perform a specific function in a specific place might pass its new genes to a disease-causing organism or an organism growing in a place where the new traits are harmful.

But Kingsbury, a microbial geneticist, said the spliced genes in bacteria released in the environment will not be like drug-resistance genes.

The ecologists are mainly concerned about uncontrolled spread or gene transfer in organisms that get less regulatory scrutiny before environmental release. The new rules exempt some categories of engineered organisms from the strongest review.

For example, the rules are less tough on gene transfers between closely related species of organisms that do not cause disease. Norse said the distinction between closely related and more distantly related organisms “shows either a lack of ecological insight or a bent much too strong toward the industry point of view.”

Several groups have complained that the new rules give less scrutiny to organisms with two other types of genetic changes: organisms that have genes deleted, but no newly added genes, and organisms that have new “regulatory” genes. These are genes that control other genes--turning them off and on, for example--but that do not code for any new proteins.

The Office of Science and Technology Policy regulations took effect June 26, but the panel that developed the guidelines will be considering comments like Cavalieri’s and proposed changes in the regulations through Sept. 26.

Rifkin has taken the authors of the new rules to court for not doing a study on the potential impact of some of the new definitions and for violating the federal administrative procedures act by not considering such alternatives as legislation.

Kingsbury and some industry spokesmen see harm in the continuing legal hassles. Kingsbury believes some animal vaccines may have been held up because “people don’t want to deal with these crazy lawsuits.”

One vaccine that Kingsbury believes will be delayed is a live oral vaccine for rabies that could be laced into food and put out for wild animals.

The Centers for Disease Control has seen dramatic outbreaks of rabies during the last eight years, with 3,040 cases reported so far this year. Most of the rabid animals have been wild raccoons and skunks in the Midwest and mid-Atlantic states. The animals are notoriously hard to capture and inoculate with existing vaccines.

“Here’s an example where the benefits of genetic engineering are enormous, and the risk of releasing the virus minimal,” Kingsbury said. “It’s an interesting test case. How long will people wait for something this good?”

Dino Dina, the director of virology for Chiron Corp., a biotechnology firm in Emeryville, Calif., said his organization has deliberately chosen to work on genetic engineering projects that would not result in environmental release of live microbes.

He said regulatory delays, safety precautions and insurance liability made it more economical to concentrate on compounds that altered bacteria could produce in the lab.

Other biotechnology industry analysts do not think the lawsuits and regulatory problems have set back the industry.

Richard Godown, executive director of the Industrial Biotechnology Assn., which represents 50 biotech companies, admitted that though “the new regulations are not everything we want,” they are “strict but workable.”

He said that with a few exceptions companies “haven’t been discernably affected by negative legal decisions” or the regulations.

Open-air testing of altered plants has begun in Wisconsin and North Carolina.

Allen Dines, director of business development at Agracetus, said his company started a field test of genetically engineered disease-resistant tobacco in late May.

“I am aware of no public concern reported over this test,” he said.

One reason, some scientists say, is that plants usually stay put, whereas ecologist Regal noted, “It’s hard to contain microbes. The wind blows them everywhere.”