Dennis Schmitt, a veterinary reproductive specialist from Missouri State University, guides an endoscopic camera inside Alice the African elephant as assistant Kristy Marson, right, and elephant keeper Brett Haskins, bottom left, assist.

ROCKARHO PUBLISHING LLC, VICTORI / VICTORIA AROCHO

PROVIDENCE — Hope for the future of African elephants arrived by air freight on Saturday afternoon at the Roger Williams Park Zoo.

Marked “Rush — Fragile,” “Biological Material” and “Do Not X-Ray,” the blue cooler contained three packages of perishable semen, collected just hours earlier from a bull elephant in Pittsburgh.

The hope rests in some small part with Alice, a nearly 4-ton pachyderm with eyes the size of billiard balls and giant flaps for ears. At age 20 or thereabouts, Alice is of prime breeding age and currently the best candidate of the zoo’s three elephants for reproduction.

Dr. Dennis Schmitt — a world expert on elephants who flew here from Missouri — examined several drops of milky fluid under a microscope at the zoo’s veterinary hospital. The sperm’s viability diminishes by the hour, and Schmitt was not elated by what he saw.

“It’s not great. But it’s acceptable,” said Schmitt, noting that the sample has enough motility to give Alice a chance at conception.

And as any biology student could tell you, all it takes is one strong swimmer to fertilize an egg.

Schmitt repacked the samples in the cooler and handed it to Tim French, deputy director of animal programs at the zoo, who rushed it to the cavernous barn where Alice waited for a carefully choreographed event nearly two years in the making.

Alice, freshly scrubbed and hosed down until her wet wrinkled hide turned gun-metal gray, stood chained at the front ankle and with her rear legs restrained in ropes. To her right stood Ginny and Kate, the other two elephants in the zoo’s tiny herd.

A team assembled: Schmitt, his assistant and four keepers. Schmitt stood in front of a cart on wheels that included a video screen, yards of flexible tubing with a tiny video camera on one end, four plastic syringes, a catheter and a jar of pink pig extender that is added to the sample to increase its volume.

Saturday marked the first of what Schmitt, French and other zoo staff hoped would be three attempts during Alice’s short window of opportunity for conception. Blood work pinpointed and an ultrasound confirmed that she was on the cusp of ovulation.

If the procedure works, Alice could give birth to a 250-pound-plus calf in 22 months.

That is, if it works.

These are tough times for African elephants. Habitat destruction and — to a lesser extent, poaching — account for their threatened status in the wild. Until recently, breeding in captive populations in zoos or parks has been difficult at best.

“Since 1980, the wild African population has fallen from 1.6 million to less than 500,000. That’s an average loss of 100 elephants every single day for the past 25 years,” says Steven Feldman, spokesman for the Association of Zoos and Aquariums.“By contrast only about 50,000 wild Asian elephants are scattered across several countries. They are endangered, but the African elephants are threatened. So looking towards the future,” says Feldman, “we are trying to conserve both captive and wild populations.”

Under Schmitt’s direction, this weekend marked the Roger Williams Park Zoo’s first attempt at artificially inseminating an elephant, a method that in 2002 produced the first two elephant calves born in captivity in the world, at the Toledo Zoo.

Artificial insemination of elephants relies on blood sampling and charting the cow’s cycle (roughly 14 weeks), and blood tests that measure the luteinizing hormone released into the bloodstream, which triggers ovulation.

A female elephant produces that hormone in two peaks that can pinpoint ovulation, said French.

“Luteinizing hormone is really the key to what has made artificial insemination in elephants successful and possible,” said French. And with only about 250 African elephants in the captive population in the country, “the current focus of the African elephant species survival plan is to make babies before too many more of these females get to that 25-year-point,” when their chance of conceiving starts to diminish.

Six to eight years ago there was little to no breeding at all, said French, and unless that changes, the captive population “will be biologically extinct” in several decades.

“There won’t be enough animals that can breed — there won’t be enough males and there won’t be enough females who are a viable age. So you’ll have animals (in captive populations) for another 40 to 50 years, but without animals coming in from the wild, there won’t be any more in zoos.”

Before artificial insemination, natural breeding represented the only means for producing elephant calves in captivity, a process fraught with the vagaries of nature and unpredictability of elephant courtship.

Typically, elephant cows were trucked to zoos with viable bulls, or vice versa. If the cow spurned the bull’s advances — or the bull showed no interest — then all was for naught.

Artificial insemination requires a keeper to manually collect the sperm, a treacherous task. As Feldman of the zoo association says, elephants are dangerous to begin with “and an excited elephant is very dangerous.”

Further complicating matters, only four bull elephants in accredited zoos are deemed viable: in Pittsburgh, Indianapolis and two in Florida. And no method has been developed for freezing and thawing elephant sperm.

“Right now we have to do all these procedures with fresh semen,” said French. “If they’re collecting it in Florida or Pittsburgh or Indianapolis, you have to get it Fed-Exed here and you have to do your insemination procedure in a matter of hours from collection. It makes it lot more difficult. If you could just go the freezer and thaw it out, you could do it when it’s convenient.”

French said some 40 zoos throughout the country, including Roger Williams are making “significant investment” toward breeding by enlarging their elephant spaces and improving the quality and complexity of those environments, said French.

On Saturday morning, the first batch of sperm missed its flight from Pittsburgh to Providence.

The staff grew palpably tense while waiting for the refrigerated sample to be flown from Pittsburgh to Boston, then driven from Logan International Airport to Providence.

At 3:30 p.m., French’s two-way radio crackled.

“There’s a package at the gate.”

The team assembled, including Schmitt; his assistant, graduate student Kristy Marson; lead elephant keeper Jennifer Warmbold; and keepers Brett Haskins, Lisa Ruggiero and Tom Troy. In the wings stood French and chief veterinarian Cheryl Cullion.

The team was relying on training that all three elephants have been put through repeatedly since summer to keep them docile and still during an ultrasound, and eventually, the insemination process. Treats, ranging from hay to slices of pumpkin, apples, carrots, green peppers, popcorn and jelly beans, helped get them through the process, with a jackpot bucket full of treats at the end..

An hour or so later, Haskins knelt on the ground beneath Alice’s hind legs and inserted the flexible tubing tipped with a video chip that allowed Schmitt to visualize Alice’s reproductive tract.

Throughout the procedure, Warmbold stood next to Alice’s head and issued constant encouragement and treats.

“Good girl. Good girl, Alice! Good girl.” “Steady, Alice!” “Steady.”

Over the loudspeaker, Sarah Brightman sang “All We Are is Dust in the Wind.” Warmbold said the music keeps the animals calm.

At 5:20, someone announced “They’re in!” as the tube entered Alice’s reproductive tract.

Only once did Alice back up and bump into the rolling cart, when some fluid spilled on the floor and she caught wind of an unfamiliar scent in the barn.

“Forward, Alice,” said Warmbold. “Steady.”

French rushed out with pineapple chunks, then popcorn and Cheerios.

Thirty-seven minutes after the procedure began, Schmitt was not entirely happy.

He had trouble visualizing key parts of her reproductive tract on the video. And the sample had only 15 percent to 20 percent motility.

If Saturday offered slender hope for conception, Sunday offered none. By the time semen from a bull elephant in Jacksonville, Fla., arrived in Providence, all it took was one look under the microscope for Schmitt to tell that it was non-viable.

Yesterday, anxiety heightened throughout the day as word came that another sample from Jacksonville was on its way.

Late in the afternoon, word came that the sample had missed the flight. And then word came that it was on the flight.

Another cooler arrived at 7:30 p.m., and by 7:45, Schmitt had more bad news for the staff.

No living sperm.

Despite the disappointment, French and Schmitt said it is still possible that Alice could become pregnant.

“It’s not unusual the first time — for me to learn her anatomy it will be easier for me to do the next time.” And the next time could be the next cycle, about four months from now.

“The more global thing is that we’re still very dependent on two or three facilities with males collecting viable sperm that day, so it points out the critical need for semen freezing techniques viable for elephants. Then the semen’s there when you need it,” Schmitt said.

French said, “My feeling all along has been that whether she conceives or not, if we have done what we can do on our end, we succeeded. And I feel we did that.” The elephant crew “did an incredible job in preparing Alice for the procedure. We made all the correct predictions from all her blood work to pick the right weekend for this. All the variables we could control, we did what we had to do. Everybody did their part and did it well.”

That included Alice, who stood quietly for 37 minutes, getting antsy only at one point.

In four months, the staff will know whether Saturday’s insemination took and whether there will be a calf in her future.

If not, said Schmitt, it will be time to try again.

kziner@projo.com