Standard Operating Procedures

Xenopus Husbandry and Care

Last updated: April 25, 2013

I. Purpose
This standard operating procedure is to provide instruction in the appropriate handling and care of Xenopus frogs

II. Responsibilities
It is the responsibility of all LAR personnel to understand and practice the proper husbandry and care of Xenopus adults housed in the animal facility

III. Housing

• Frogs are housed in Health Sciences Building (HSB) 018A
• Upon entering 018A put on designated lab coat and gloves
• Frogs are housed together in a gray Rubbermaid pool
• The pool has a 50 gallon capacity; add approximately 20 gallons of treated well-water to pool (see Sections IV and V)
• Eight is the maximum number of frogs for the pool
• Pieces of 7" by 2-1/2" PVC pipe and synthetic lily pads are used for enrichment and hiding places

IV. Water

• Use well-water from room HSB015
• Fill 2 5 gal Nalgene water buckets and let water stand for at least 24 hours before using to de-gas the water and to bring to room temperature.
  • Well water should be chlorine and choloramine free
  • Bring water to room temperature (70^o F) to avoid temperature shock to the frogs (a 1 or 2 degree change in temperature can be detrimental to the frogs)
  • A water thermometer with a probe in the tank automatically measures the water temperature and gives a digital readout
  • The correct water temperature for adult Xenopus is 68-75^o F
• Add appropriate amount of Lymnozyne to water (see Section V)

V. Lymnozyne

• Lymnozyne has a biological formula of naturally occurring bacteria, enzymes and micronutrients to help prevent disease in frogs. Lymnozyne helps reduce levels of Aeromonas and Pseudomonas bacteria which can cause red-leg and other gram positive and negative bacterial infections 
• To use: mix 1 mL of undiluted Lymnozyne (from original bottle) with 10 gallons of well water
• Let treated water stand overnight to de-gas and to bring to room temperature (use thermometer)

VI. Handling the frogs - always wear gloves.

• Handle the frogs as little as possible to avoid causing them stress
• Best practice is to use a net to catch and handle the frogs
  • Scoop frog out of water using net.
  • Place free hand over top of net to prevent frog from jumping out
  • Netting prevents damage to the mucous layer on the frog and is a less traumatic means of handling
• To restrain frog using your hands:
  • Gentle handling is essential for minimizing stress to the frogs
  • Hold the frog's head toward the handler's wrist.
  • Grasp frog palm down while placing fingers on each side and between legs
  • Gently but firmly secure frog with both hands

VII. Daily, water pH, siphoning and observations. Always wear gloves.

• Test the pH of the water in the tub and record it in log book
• Observe frogs for signs of disease, illness, or injury and count frogs.
• Siphon the water from the pool using the rubber siphon tube
• Leave frogs in the pool to lessen their stress
• Leave about an inch of water in the pool (approximately 5 gallons
• Retrieve as much debris as possible using the siphon and net
• Refill frog pool with 2 5 gallon buckets of aged well-water and Lymnozyme
• Replace clear plexiglass covering over pool and place heavy rock on lid for security
• Fill out daily checklist and census sheets

VIII. Every 2 weeks, cage washing. Always wear gloves.

• Gently transfer frogs to rat boxes filled with treated water and cover boxes with a rat box lid so that frogs cannot escape. 
• Empty water from gray Rubbermaid pool
• Wash pool, enrichment devices, etc. by hand with well-water and a scrubber
• Rinse pool and accessories twice with well water and discard the rinse water
• Fill pool with treated water from 2 5 gallon buckets if treated and aged well-water
• Gently transfer frogs back to cleaned pool and replace lid and heavy rock

IX. Surgical Aftercare

• Following surgery, place frog in a rat box with treated water from frog pool and cover with a rat box lid so that frog cannot escape.   Post-op frogs will be properly identified with surgery date, return to tank date and ID#
• Monitor frog for 72 hr following surgery and log all observations in post-op log book.
• Return frog to main pool if it appears healthy at the end of the 3 days.

X. Feeding

• Feed frogs with Xenopus Express Floating Frog Food.
  • Large size pellets
  • 3/16 in
  • 45% protein
• Feed frogs every afternoon.
• Feed each frog two pellets of frog food.
• Remove left over food the next day when doing the daily cleaning and observation.
• Report failure of post-op frogs to eat to AV.

XI. Frog procurement

• Frogs are supplied by Xenopus Express.
• Frogs are shipped in sealed containers with moist foam cubes to prevent desiccation and provide protection.
• Check box for damage immediately upon arrival.
• Unpack box carefully to avoid cuts or injury to frogs.
• Take digital photos of dorsal side of individual frogs for identification purposes.  Post  laminated photos with assigned frog number on HSB 018A
• Rinse frogs with treated water.
• Place frogs in gray Rubbermaid pool.
• Record arrival conditions, etc. on daily checklist and on all other shipment/procurement documents.
• Contact PI to report arrival of shipment
• File all paperwork in HS009 file cabinet in 'Order Form' file

Always wear gloves when working with frogs.

Always wash your hands after working with frogs.