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Shipping/Receiving X. tropicalis

NOTE: It has been our experience that making arrangements to ship Xenopus can be somewhat difficult. We have created this web page to help facilitate shipping and receiving frogs. However, users are warned that shipping frogs can be potentially dangerous even after following all of the recommendations listed below. Unforeseen factors can always impact the health of frogs being shipped. We hope to continue to update this page with user comments and our experiences so that we can create a protocol that maximizes the health and well-being of shipped frogs.

Receiving

You can expect frogs to arrive at your facility in various types of packages.

There is usually a cardboard outer box, often with a solid Styrofoam inner container, or packing peanuts. Frequently, holes are poked through the lid of the Styrofoam container, and the cardboard. This type of package works well because it protects the frogs from sudden temperature changes, as well as offering some physical cushion.

Inside, the frogs may be packaged in individual containers (“GLAD” food containers, for example), plastic bags, or simply loose in the box or Styrofoam container. Be prepared for anything, and use caution when opening all the layers of the package. Tropicalis squeeze into the smallest crevices and can be tricky to find. An inventory of the frogs you are receiving is critical to be sure they are all accounted for.

There is usually some wet material to keep the frogs moist, such as damp sphagnum moss or sponges. There may be water with the frogs. We have seen a case where the veterinarian advised packaging the frogs in so much water, the airport refused to accept the leaking packages. This much water is unnecessary. Frogs do fine with a small amount of water in moist sphagnum moss or damp sponges. Read the following “Packaging” section for further details.

If the frogs are shipped with sphagnum moss, it is a good idea to have an intermediate container for rinsing the moss off before placing the frogs in their new tank. This will avoid getting dirt and moss in the tank and drainage system.

If the frogs have been delayed, or if the packaging is inadequate, the frogs may suffer from dehydration and temperature fluctuations. Get them into shallow water of appropriate temperature as soon as possible, and observe them carefully. If necessary, contact a veterinarian.

Quarantine

We generally quarantine frogs for one month after arrival to our facility. We keep them in the same room with our other frogs, but house them on a separate, isolated row of tanks. The water is not shared with any other frogs. Frogs can be ovulated two weeks after arrival, provided they are healthy.

The frogs should be fed the day following arrival in your facility. Nasco Frog Chow or HBH Frog and Tadpole Bites are two diets we commonly use. The HBH is moist and often more readily accepted by a stressed frog. If the new arrivals are housed two or more to a tank, competition may cause them to eat more readily.

If the frogs arrive well fed and healthy, feed a normal ration of food for each frog. Supplement with extra feedings if the frogs seem thin, especially when all the food is immediately eaten. A large supplier sent us frogs, which seemed thin and quite hungry. These frogs were poor egg producers. We fed these frogs as much as they would eat for a month, two to three times a day, until they gained weight and began to lay good amounts/quality of eggs.

 During the quarantine period the frogs are carefully observed for illness, abnormal behavior, and weight loss each day. There are a number of diseases affecting X. tropicalis. A latent infection may become acute due to the stress of travel. Read the Diseases section for descriptions of various illnesses. Notify a veterinarian if you observe anything abnormal.

 

We have had success in treating stressed tropicalis using a salt solution in place of regular water. In one instance a frog arrived and lost its ability to osmoregulate. It became bloated and was floating at the top of the tank. We placed it in a salt solution and it recovered slowly over the next few days. A solution of rock salt (1g/L) or Holtfretter’s solution can be used. Keep the frog in the solution until it recovers. Change the solution daily.

Shipping

Packaging

There are many different methods used to package X. tropicalis for shipment. Please refer to the Live Animal Regulations (LAR) for standard guidelines on shipping animals. The LAR is published yearly by the International Air Transportation Association. The regulations contain information on appropriate shipping container sizes and animal density. The guidelines can be purchased on the IATA website. Also, AALAC has a good article on the general considerations for preparing to ship animals, such as applicable regulations and proper documentation. This article has links to numerous useful “regulatory and oversight organizations” which may affect your shipment, such as the USDA and U.S. Customs.

We generally do not ship more than 6 – 10 frogs at a time. We use an outer cardboard container, with a Styrofoam container fitting snugly inside. (Picture 1, 2). These are readily available in various sizes from our shipping/receiving department. You may find your shipping department is more than willing to give you these containers that might otherwise be discarded.

Picture 1: Styrofoam box fit snugly inside cardboard box.

Picture 2: Styrofoam box with lid on

 

The frogs are packaged inside GLAD or Ziploc containers (Picture 3, 4). The GLAD Medium Soup and Salad (3 cups), or the Ziploc Medium (4 cups) Square are good for 4 frogs. For 1 or 2 frogs, the Ziploc Small (2 1/2 cups) Square would work. Putting the frogs in the plastic containers allows you to separate different types of frogs and label them. Putting frogs directly in the Styrofoam is not recommended, as the abrasive surface can damage frog skin.

 

Picture 3: “GLAD” container used to hold frogs.

Picture 4: Side view of container

 

First make four slits with a utility knife in the lid for air by simply inserting the blade into the lid. There is no need to cut out a hole. Before adding the frogs, sphagnum moss (we get it at Ace Hardware but can be available at plant nurseries, hobby shops, etc) that has been soaked overnight in water is put in the plastic container. We also add about 50 ml. of water, to ensure the moss does not dry out. After adding the frogs, put in some more moss to cover the frogs. They will quickly burrow in. Follow a rule of “thirds” when packing: one third of the plastic container should be moss, one-third should be airspace, and one-third frog mass.

Carefully mark each container with the contents. We usually mark directly on the plastic with a permanent marker. Several rubber bands are wrapped snugly around the plastic container, to ensure the lid does not come loose.

The plastic carriers are placed in the Styrofoam container, using newspaper to keep them from moving around. Use up to 2/3 of the space in the box for the plastic containers, and keep 1/3 for airspace/ packing material. The Styrofoam lid is sealed to the bottom with shipping tape. The cardboard box is then sealed. We do not make additional air holes into the Styrofoam or into the cardboard box. In our experience, by following the rule of thirds, the frogs have plenty of air inside the box. This would obviously be unacceptable for small mammals since they have considerably higher metabolism rates and greater oxygen requirements. However, it has been our experience that holes often lead to leaking containers. These wet and leaking containers are then unacceptable for continued shipment and can get delayed in transit. We have had multiple episodes where frogs were injured or died due to dehydration from prolonged travel times or leaky containers.

Shipping Temperatures

If it is going to be cold, place chemical heat packs (like the ones for skiers) at the bottom of the Styrofoam box. Do not allow the heat packs to come in direct contact with the frogs, as they can harm frog skin. Alternatively, we put the ice packs that come with restriction enzymes or other molecular biology reagents at 37oC overnight. Pack them at the bottom of the Styrofoam container in the morning on the day of shipment. This should help keep the temperature in the Styrofoam container acceptable to frogs in cold weather.

We have not had a lot of experience with shipping frogs in extreme temperatures, so have rarely used heat packs. If anyone has specific suggestions for the number of heat packs to use, and the temperatures at which they should start being used, we would like to know. 10oC and 33oC are the absolute minimum and maximum recommended air temperatures for shipping frogs.

We encourage others who have shipping suggestions or experiences, good or bad, to share them. Hopefully, we can continue to update and improve shipping procedures in the Xenopus community.

 

Communication

It is important to have a contact person at your lab, as well as at the receiving lab, to coordinate the shipment. The veterinary staff at both ends will need to be informed. These steps ensure the frogs are properly received and cared for at their destination. We strive to obtain our frogs first thing in the morning when they arrive at our loading dock so we can get them into a more suitable environment as soon as possible.

 

We find it useful to maintain a database with Shipping information for each location we ship to.

 

Regulations and Shipping Forms

Although there are no required forms, we recommend looking into state code regulations, and country regulations if shipping internationally.

Please refer to the Live Animal Regulations (LAR) for standard guidelines on shipping animals. The LAR is published yearly by the International Air Transportation Association. The regulations contain information on appropriate shipping container sizes and animal density. The guidelines can be purchased on the IATA website.

Also, refer to the AAALAC article on the general considerations for preparing to ship animals, such as applicable regulations and proper documentation. This article has links to numerous useful “regulatory and oversight organizations” which may affect your shipment, such as the USDA and U.S. Customs.”

We do our shipping through UC-Berkeley's OLAC. Important information needed for shipping can be found on the Shipping Request form.

 

Timing Shipments

Frogs should be ready for courier pickup by 10 am the day of shipment. Shipments should be scheduled only on Monday through Wednesday to avoid having frogs delayed in transit on the weekends. Be aware of holidays.

 

Carriers

This is a list of carriers who will accept frogs, in our experience.

Menlo Worldwide  

Bax Global

FedEx - Animal shipments must be coordinated through the Live Animal Desk. See their terms and conditions for more information. NOTE: We had an experience where FexEx would not deliver to a remote location.

 DHL

Maura Lane, Kendon Kuo, Mustafa Khokha, Tim Grammer