Wildlife Conservation: Breeding Eastern Tiger Salamanders

Amphibian breeding season is here! That means it’s time to start helping amphibians get in the mood for love and romance.

The Institution of Museum and Library Sciences (IMLS) recently awarded a $500,000 National Leadership Grant for the purpose of improving reproduction within captive assurance colonies of imperiled salamanders. The Detroit Zoological Society is one of the primary partners on this grant. My doctoral research focused on salamanders, their reproductive physiology and techniques to help them breed, and that very research was the basis for the recent IMLS grant proposal.

Other partners on the grant include Mississippi State University and Omaha’s Henry Doorly Zoo and Aquarium. Now that the grant was awarded, techniques must be taught to the other partners so we can all work together in salamander conservation efforts. I recently visited Mississippi State University in order to train the other principal investigators and the graduate students involved on this salamander grant in principles of natural salamander reproduction and performing assisted reproduction techniques with salamanders.

The training session involved eastern tiger salamanders (Ambystoma tigrinum), which are regionally threatened and endangered in various areas of North America. Like many species of salamanders, the tiger salamander is very difficult to breed in human care. Similar to other species in the “mole salamander” family, tiger salamanders respond to changes in air pressure and temperature when seasonal rain storms occur. These storms are what cue breeding behavior and are very difficult to replicate in human care. Without the ability to provide the appropriate “mood”, tiger salamanders in human care will not often feel romantically inclined. Natural breeding is always the first goal when breeding animals for conservation, but sometimes this is extremely challenging. In these cases, we use alternative techniques while we perfect replicating the natural breeding environment.

In vitro fertilization is a technique used to assist salamanders and other amphibians in breeding. Most salamander species undergo internal fertilization, in which the female picks up a capsule of male sperm, called a spermatophore, which the male has deposited into the environment. The female holds the sperm in an internal pouch that she later empties over her eggs as the eggs are laid. In in vitro fertilization of salamanders, sperm is collected from males by giving them a massage. Eggs are then collected from the female into a small dish where sperm is placed on top of the eggs. Just add water, and presto, you have salamander babies. Of course, it is never just that easy, but the concept is straightforward.

I trained the other primary investigators and students in other techniques as well, including cryopreservation – or freezing and long-term storage – of salamander sperm and sperm quality assessment. The training was very successful, with nearly 100 tiger salamander babies produced. Now the trainees can go on to teach more amphibian conservationists, and we can save more species by assisting with breeding!

– Dr. Ruth Marcec is the director of the National Amphibian Conservation Center at the Detroit Zoo.

Wildlife Conservation: Lending Amphibian “Sexpertise” to Other Accredited Zoos

I recently participated in a partnership to save an incredible species of amphibian: the blue spotted salamander (Ambystoma laterale). This animal is regionally endangered in parts of the Midwest and about 10 years ago, the Iowa Department of Natural Resources started a partnership with Omaha’s Henry Doorly Zoo. They wanted to breed these salamanders for release at locations where they once thrived but have now disappeared. Having had little success in doing so thus far, the organizations reached out and requested my amphibian “sexpertise”. I travelled to Omaha to deploy some of my special reproductive techniques with the blue spotted salamander assurance colony living there.

It comes as no surprise that these salamanders proved very difficult to breed. Blue spotted salamanders need an extraordinary amount of “salamander romance” in order to breed naturally. These animals undergo winter brumation (a hibernation-like state). In the spring, they will emerge from brumation when it begins to rain and when the temperature rises in order to migrate to breeding ponds. Their migrations can be over several miles, and most salamanders return to the same pond every breeding season. In captive breeding situations, we attempt to recreate the cues that the salamanders take from the wild in order to get them “in the mood” for breeding. We cool them down and offer them the opportunity to undergo brumation. We place them in rain chambers so they can experience an “indoor storm”. We also attempt to create a naturalistic pond for them to breed in with proper vegetation and leaf litter.

Sometimes, despite our best efforts, the natural cues we provide artificially are not enough to trigger breeding. Many amphibians are having difficulty breeding in the wild, due to changes in the climate and their habitat. Given that the natural environment is not currently providing ideal cues for breeding, it is especially difficult to perfectly recreate environmental breeding cues for animals in human care. When captive animals do not breed in response to the cues we provide, we often use hormone treatments to give them a little boost. These hormone protocols are meant to trigger breeding behaviors as well as the release of sperm and eggs. In the best-case scenario, the application of hormones will result in the animals breeding on their own. In the worst-case scenario, the animals will not breed on their own after the application of hormones and an in vitro fertilization (an artificial fertilization of the eggs with the sperm in a petri dish) will take place.

We used a combination of natural cues and hormones to stimulate breeding in the blue spotted salamanders. The amount of hormones I gave to the female blue spotted salamanders was determined by how developed their eggs looked on ultrasound. While giving the animals hormones resulted in many interesting behaviors, the salamanders did not breed on their own. After giving them time to breed on their own without success, I attempted in vitro fertilization. In order to perform this technique, I needed to collect eggs from the females and sperm from the males. There was one big problem with attempting this technique – the male blue spotted salamanders were not producing sperm. Fortunately, I had a back-up plan!

The blue spotted salamander is no ordinary salamander. In the wild, many female blue spotted salamanders breed through a special adaptation called kleptogenesis, meaning that they steal sperm. You read that correctly: sperm thieves. Most salamanders undergo internal fertilization. This occurs when the male deposits the sperm in a ball – called a spermatophore – into the environment. Through a courtship dance, the spermatophore is collected by the female. These special female blue spotted salamanders will find and collect spermatophores deposited by males from other, closely related, species (including Jefferson’s salamanders, tiger salamanders, and small-mouth salamanders). This is the part where it becomes complicated. Rather than using the sperm to fertilize their eggs and have hybrid offspring, an enzyme in the sperm activates the egg which begins to grow into an embryo. The rest of the sperm is discarded, and the offspring which develop are essentially clones of the mother. Knowing that blue spotted salamander females are often sperm thieves, I had the Omaha Zoo’s resident male tiger salamanders (Ambystoma tigrinum) on standby. I was able to collect sperm from the tiger salamanders in order to use in the in vitro fertilization of the blue spotted salamander eggs.

Unfortunately, the eggs did not fertilize with the tiger salamander sperm. While we did not achieve offspring, we learned what we need to do differently next time around. We certainly will not give up on these amazing animals. We will try to breed the blue spotted salamanders again later this year, using slightly different techniques.

Thank you to our partners at Omaha’s Henry Doorly Zoo and the Iowa DNR.

– Dr. Ruth Marcec is the director of the National Amphibian Conservation Center at the Detroit Zoo.

Amphibian Conservation: Fired Up About Salamanders

From the days of old, salamanders have captured the imagination of humankind. Long ago, when people would place logs in a fire, brightly colored salamanders would often come scurrying away from the flames. Of course, this was because the salamander’s hiding place was burning, but people in medieval times assumed that the salamander was a creature that came from the flames. People supposed that salamanders were imbued with mystical properties and they became the alchemic symbol for the element of fire.

Despite this long-time association with flame, salamanders are unlikely to be found near a toasty fire. There are more than 600 species of salamanders known, and most of them prefer cool, damp places to live. These critters are often found in a variety of places, including under logs, in rock crevices, high in trees, or living in cool bodies of water, ranging from puddles to fast moving rivers.

Unlike lizards, which look similar to salamanders, salamanders have moist, scale-less skin. Salamanders use their unique skin to intake water, absorb nutrients, and even breathe.

Most species of salamanders are born with a larval stage, similar to the tadpole stage of a frog. The larval salamander is aquatic, has external gills, and has no limbs upon hatching. As the larva grows, it develops legs. Many species of salamanders undergo metamorphosis, losing their gills and undergoing other small body changes as they transfer from water to land.

Salamanders are critical indicators of environmental health, and are key players in the health of ecosystems. These animals serve as sentinels for anything that is changing in the environment. Such changes include pollution, disease or climate change. Almost 50 percent of salamander species are threatened with extinction, according to the International Union for Conservation of Nature (IUCN). The Detroit Zoological Society is working toward preventing these wonderful creatures from going extinct through breeding and release programs, habitat restoration and population surveys.

We invite you to celebrate these remarkable creatures with us during a special event called Salamander Saturday on May 6. National Amphibian Conservation Center staff and volunteers will provide informative zookeeper talks, storytelling, games and crafts. This event is free with Detroit Zoo admission. Sneak a peek at salamanders and learn all about these unique animals during this fun family event.

– Dr. Ruth Marcec is the Director of the National Amphibian Conservation Center at the Detroit Zoo.