About
Axolotls...
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by Susan T. Duhon
The Indiana University Axolotl Colony was founded in 1957 by Rufus R. Humphrey,
who brought his small research colony to Bloomington when he retired from
teaching at the University of Buffalo. The small colony gradually grew and
became a genetic resource center as Humphrey pursued his research interests
in mutant genes of the axolotl. Since 1969 the colony has been supported
by the National Science Foundation as a living stock center to provide research
material for research and instruction to laboratories and schools throughout
the United States. Today we breed axolotls about ten months out of the year
and send out tens of thousands of embryos, larvae, and adults to developmental
biologists, neurobiologists, and other amphibian research scientists, as
well as to primary, secondary, and undergraduate-level teachers. This guide
is a partial response to the frequent questions and requests for advice
and information on the care and feeding of axolotls that the colony receives.
Water is the most important component of the axolotls' environment. Never
house them in extremely soft or distilled water. They need hard water
to help them maintain the integrity of their skin, their most important
defense against infection. Remove any chlorine, chloramines, or ammonia
that may have been added as part of municipal water treatment. Commercial
preparations (e.g., Amquel) are available for this purpose.
We pass our water through an exchange resin to remove heavy metals
and past a sterilizing uv light as well. We also add salts to the
water to make a modified Holtfreter's solution.
The recipe we use to make 40% Holtfreter's in a 44 gallon barrel is:
KCl: 1 teaspoon
CaCl2: 2.5 teaspoons
MgSO4.7H2O: 2 tablespoons
NaCl: 240 ml (dry but measured in a liquid beaker)
More on salt solutions
The salts restore hardness after water treatment and help
us maintain the animals' health by discouraging parasites and fungus.
Extra salts are not essential, however, if you are attentive to good husbandry
practices and the water is hard and free of chemicals and heavy metals.
Keep the pH between about 6.5 and 8. If pH is at the high end of this
range, monitor ammonia carefully because its toxicity will be greater
than at neutral pH.
We house adult axolotls either individually in one gallon bowls (either
squat, glass fish bowls or plastic ice cream buckets) or in groups of three
to four in plastic tubs, approximately 10" x 18" x 6" deep (25 cm x 46 cm
x 15 cm). A half gallon (2 liters) of water per axolotl is adequate, provided
that the animal is completely submerged and the water is changed frequently.
We prefer to keep male axolotls in individual bowls when they are not being
mated because they breed better for us when housed singly between breedings
than they do housed in groups. Female axolotls spawn well for us whether
housed singly or in groups.
Although we use individual containers or small-group containers, others
have successfully housed axolotls in filtered aquaria. As a rule of thumb,
house two adults in a ten-gallon aquarium. Gravel, if used, should ideally
be coarse. The axolotls will ingest smaller gravel, especially pea-sized.
Although they regurgitate the gravel eventually, it's easier to avoid
the problem. Be careful, especially if you use a power filter, not to
circulate the water too fast (see below) and to maintain a biological
filter that can convert ammonia and nitrites. Do partial water changes
regularly and keep the water cool and out of direct sunlight.
You may find it difficult to raise small larvae in an aquarium because
they will be very aggressive and cannibalistic toward one another. Juveniles
and adults, however, can thrive in a properly maintained aquarium.
Axolotls in nonfiltered systems don't need continuously circulating water
if the water is replaced regularly. In fact rapidly circulating water
is stressful to them. If your system circulates water continuously, keep
the rate of circulation as slow as possible. Nor do they seem to need
a large volume in which to swim and move around. Even when placed in an
aquarium, they spend the bulk of their time lying nearly motionless at
the bottom.
They are extraordinarily resourceful when it comes to gas exchange. If
you watch an axolotl for several minutes you will see it flick its gills
periodically. Moving the fronds back and forth quickly stirs up the surrounding
water and disperses any carbon dioxide that may have accumulated about
the gills and mixes in fresh, oxygenated water. You may see the axolotl
swim to the surface and gulp air as well because, although outwardly the
neotenic axolotl has a larval form, it still undergoes "cryptic" metamorphosis
and develops lungs.
Although we successfully use plastic containers, they must be kept very
clean. Plastic tends to support the growth of a bacterial scum along the
bottom and sides of the container. We have found that if this scum is
not cleaned off periodically, the axolotls will develop sores on their
toes and feet that will not heal. Instead the skin retreats up the leg
and the toes eventually die and are sloughed off. If this happens put
the axolotl temporarily into a glass bowl. Add a few drops of mercurochrome
(enough to tint the water a pale orange) to the water as a disinfectant,
and change the water frequently. The axolotl will heal nicely, and the
toes will probably regenerate.
We keep young, larval axolotls in shallow glass bowls about 8" (~20 cm)
in diameter. When they first hatch, fifty or more larvae may occupy the
same bowl, but as they grow, reduce the number per bowl progressively.
The larvae's rate of growth depends upon temperature, frequency and amount
of food, and the number of animals per bowl. Larvae should be about an
inch long by the time they are 1 1/2 to 2 months old.
Axolotl larvae never all grow at the same rate, so when it is time to
divide them up, be sure to put similarly sized animals together. If mixed
sizes are housed together, larger ones will try to eat the smaller ones.
In any case, any young axolotls housed together will tend to lack toes
or feet, because during this phase of rapid growth, the larvae will snap
at anything that moves. For this reason, we try to put all young axolotls
2" (5 cm) or longer by themselves. We use small plastic bowls containing
one liter of water for this purpose. Make sure the sides are tall enough
to keep the axolotls from jumping out. If any larvae have lost toes to
their fellows, they will regenerate them quickly.
Axolotls thrive at cool temperatures. We keep our axolotls at 15-18°C
(60-65°F). They should never be kept above about 22°C (72°F).
Too warm temperatures are dangerous for axolotls. To prevent overheating
never house them where they are exposed to direct sun.
In their native habitat, axolotls ate of the abundant small fauna, including
snails, worms, crustaceans, various small invertebrates such as Daphnia,
and small fish and amphibia (Shaffer, 1989). In the
laboratory we feed our young larvae brine shrimp and juveniles and adults
pelleted food.
We always change the water in the axolotls' bowls before feeding. To
change the water of axolotl larvae, carefully pour the axolotls from their
bowl into a net. Clean the bowl with scrub mixture (baking soda and salt
mixed together in a 2:1 ratio). Put one liter of clean water in the bowl,
then invert the net and release the larvae into the clean water.
Feed newly hatched, live brine shrimp
to young larvae daily. We strain the shrimp out of the brine, resuspend
them in axolotl water, and deliver them to the axolotls using a large
glass pipette. Feed enough shrimp so that all of the young larvae have
orange bellies afterward, but few shrimp are left uneaten in the bowl.
Change the water again within 24 hours after feeding, because shrimp cause
a rapid deterioration of water quality if many are left uneaten.
When the larvae get to be about 1 1/2" (4 cm) long, we begin to supplement
the shrimp diet with pellets. We use soft-moist salmon pellets, a vitamin
and mineral fortified, fishmeal-based sinking pellet, 1/8" (3 mm) in diameter
(Rangen, Inc.). We gradually wean the larvae off shrimp and increase
the number of pellets being offered until the young axolotls are 2-3"
(5-8 cm) long, when they no longer need any shrimp.
As the larvae grow, we increase the number of pellets, always seeking
to fill the larvae up without leaving a lot of extra food to foul the
water.
Young larvae are very vulnerable to disease, but difficult to treat.
Therefore, we have found the most effective strategy is to feed them generously,
pay particular attention to water quality, guard against overcrowding,
and grow them up out of the vulnerable stage as quickly as possible.
Always change the water of pellet-fed axolotls before feeding. We change
young animals' water daily, and adult animals' water at least every other
day. Frequent changes prevent the buildup of ammonia and other metabolic
wastes and keep a large population of bacteria from getting established.
We change the water of axolotls housed in individual bowls by carefully
pouring it, axolotl and all, into a plastic colander. We clean the bowl
with scrub mixture (baking soda and salt mixed together in a 2:1 ratio),
put in a fresh pitcher of water, and return the axolotl to its home. The
axolotls will tolerate short periods of time out of the water very well.
They are able to continue gas exchange as long as their skin is moist,
but never leave them out of the water long enough for their skin to dry
out or become "tacky."
We pump water from a large tank to flush the tubs of the axolotls housed
in groups. Prior to flushing, we vacuum up (by aspiration) uneaten food
and feces from the bottom of the tub. Once a month we transfer the animals
from their tubs to a temporary holding tank and clean off any scum that
has accumulated on the sides and bottoms of the tubs.
When axolotls reach about 6" (15 cm) in length, we begin feeding them
3/16" (5 mm) diameter pellets. When they are about one year old, decrease
the number of feedings to three or four times per week, no more than five
pellets at a time.
Axolotls may snatch pellets dropped in front of them out of the water,
using their lateral line to detect the falling pellets. They also quickly
learn to locate those pellets that fall to the bottom of the bowl. Often
you can see them arching their necks as they move about their bowls seeking
food that is on the bottom.
Juvenile and adult axolotls will accept a wide variety of foods. Besides
salmon pellets, other feeding possibilities include Daphnia or
water fleas, mosquito larvae, earth worms (a favorite), "feeder" guppies
or goldfish, trout pellets, and beef liver or heart. Any live fish or
amphibia used as food should be aquarium bred in order to avoid the introduction
of harmful parasites. Never, for example, feed fresh water minnows obtained
from bait shops because they are usually heavily infested with parasites.
You can distinguish adult male axolotls from females by their relatively
straight bodies and large glands about the cloaca (vent). Mature females
have round, plump bodies because they are filled with eggs, and they lack
the conspicuous glands. Axolotls cannot be sexed until they are sexually
mature, at about one year.
Choose healthy, mature animals for mating and put them together in a
suitable container. We use large plastic tubs. Add enough water to cover
the animals. The bottom of the container must be textured (that is, not
smooth glass), or contain rocks to which the male can attach his spermatophores,
small cones of clear jelly with a white, sperm-containing cap. Cover the
container and leave them alone.
The male will begin to court the female by nudging her with his snout.
This begins a short mating dance, during which the male deposits his spermatophores
on the rocks. The female follows behind and picks up the spermatophores
with her cloaca and stores the sperm inside in a special structure called
the spermatotheca (Eisthen, 1989).
The next day, or at least a few hours later, look for spermatophores
on the rocks or loose in the water (The female does not usually pick them
all up). Return the male to his home.
If you see spermatophores, place the female in a glass bowl. Cover her
with a towel and wait. By the next morning, you will probably see eggs
in the bowl if she is going to spawn. Usually the female will begin to
lay her eggs 12 to 20 hours after mating. Rarely, she will wait several
days to begin. She sheds her eggs over a period of one to two days. If
the female spawns in a container that has rocks or plants, you will find
the eggs spread about on them. She never lays her eggs in a single mass
or clump. If you allow your axolotls to spawn in an aquarium, you will
need to remove either the eggs or the adults before the eggs hatch. Otherwise
the adults will eat the hatchlings. Usually they will not touch eggs while
they are still in their jelly coats.
We allow males at least a week or two and females at least two, preferably
three months, between spawns.
We never use hormones to induce spawnings, because healthy axolotls are
not difficult to get to spawn. The success of a particular pairing is
unpredictable however. Most pairings should produce spermatophores, and
at least 1/3 to 1/2 of the ones with spermatophores should produce spawns.
Changes in lighting are critical for spawning. If we shorten the light
period, we get fewer spawns. If we then lengthen it gradually over a period
of a few weeks, spawns will again increase in number. The absolute length
of the light period is not critical. Our axolotls get about 14 hours of
light each day.
In their native habitat, axolotl spawn in February (Gadow,
1903), but our captive-bred axolotls display only residual seasonality.
We obtain spawns readily year round, although we have slightly less success
from August through October.
Collect the eggs from the female's bowl with a wide-mouthed pipette. Remove
eggs attached to rocks or plants with forceps. Axolotl eggs are enclosed
in a clear jelly coat that protects them from physical injury and bacterial
infection. Eggs that accidentally come out of the jelly usually die without
special handling.
Place the eggs in a shallow bowl such as those in which young larvae
are kept. Avoid overcrowding. Put no more than 50-100 eggs or developing
embryos in each bowl. Discard any excess jelly, and do not allow the embryos
to clump. Embryos in the interior of a clump may not receive enough oxygen.
If you see embryos in the center of a cluster of eggs developing more
slowly than those on the periphery, use forceps to pull the clump of embryos
apart. Otherwise the embryos in the middle will die.
It is a good idea to sort the embryos when they are about a day old (at
room temperature they will have reached blastula stage) to remove infertile
and nonviable eggs.
You can control the embryos' rate of development by modifying the temperature
at which they are kept. Embryos that have reached blastula or later stages
may tolerate temperatures as low as 1-2°C for from one to three weeks
(Ginsburg et al., 1987), but earlier embryos may
be damaged by temperatures below about 10°C. A lower temperature
delays development, and raising the temperature (to a maximum of 25°C)
accelerates development. We often hold embryos at cold temperatures for
anywhere from a few days to a week or more so that we can ship them out
to laboratories while they are still at an early stage of development.
Embryos kept at room temperature will hatch in two to three weeks. As
the larvae hatch, carefully remove the discarded jelly capsules with a
pipette. Some larvae may not hatch on their own. After the majority have
hatched, remove the jelly coats from any unhatched larvae by gently puncturing
the capsules with forceps.
Begin feeding the young larvae when you can no longer see yolk in their
bellies. Usually they are ready to eat soon after hatching. If they are
not fed right away, they may swallow air, and get air bubbles in their
stomach. When they begin to eat, however, they will be able to expel the
air.
Axolotls that are cared for regularly, fed properly, and kept in clean water
at suitable temperatures are hardy animals that seldom get sick. Axolotls
stressed by poor husbandry, adverse environmental conditions, or experimental
procedures are vulnerable to infection by opportunistic bacterial pathogens.
Common pathogens are Pseudomonas, Aeromonas, and other gram-negative organisms.
The first signs of illness are loss of appetite and deterioration of
the gills. You may see some anemia. More severely ill axolotls may be
jaundiced and have small open skin sores. Very ill animals may develop
ascites or severe edema.
Aside from very young larvae, we have found that axolotls about one year
old that are just becoming sexually mature are most prone to illness.
Prompt treatment with antibiotics will often help. We use amikacin diluted
with physiological saline to 5 mg/ml. We give the axolotls three intraperitoneal
injections (5 mg/kg body weight) 48 hours apart. The injections are best
given at the first sign of illness (Duhon, 1989a;
1989b).
Axolotls are fascinating creatures and important laboratory animals.
Their status as an endangered species underlines the importance of proper
care and maintenance of health.
Duhon, Susan T. Diseases of axolotls. In: John B. Armstrong
and George M. Malacinski, Eds. Developmental Biology of the Axolotl.
Oxford University Press, New York, pp. 264-269, 1989.
Duhon, Susan T. Disease control in a large colony of axolotls.
Herpetopathologia 1:105-108, 1989.
Eisthen, Heather L. Courtship and mating behavior in the
axolotl. Axolotl Newsletter 18:18-19, 1989.
Gadow, H. The Mexican axolotl. Nature 1736:330-332, 1903.
Ginsburg, Mary F., Twersky, Laura H., and Cohen, William
D. Ambystoma embryo development after cold storage. Axolotl Newsletter
16:3, 1987.
Shaffer, H. Bradley. Natural history, ecology, and evolution
of the Mexican "axolotls." Axolotl Newsletter 18:5-11, 1989.
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