Nov 29, 2023

Public workspaceQuarantine and heat treatment for Hymenochirus boettgeri infected with Bd

DOI
dx.doi.org/10.17504/protocols.io.4r3l22b9jl1y/v1
  • Tamilie Carvalho1,
  • Catherine Si1,
  • Rebecca A. Clemons1,
  • Evelyn Faust1,
  • Timothy Y. James1
  • 1Department of Ecology and Evolutionary Biology, University of Michigan, Ann Arbor, MI 48109 U.S.A
Timothy James
Open access
DOI: dx.doi.org/10.17504/protocols.io.4r3l22b9jl1y/v1
Protocol Citation: Tamilie Carvalho, Catherine Si, Rebecca A. Clemons, Evelyn Faust, Timothy Y. James 2023. Quarantine and heat treatment for Hymenochirus boettgeri infected with Bd. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l22b9jl1y/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 20, 2023
Last Modified: November 29, 2023
Protocol Integer ID: 91219
Funders Acknowledgement:
Gordon and Betty Moore Foundation
Grant ID: 9337
Abstract
This protocol outlines the procedures for conducting heat treatment for clearing Batrachochytrium dendrobatidis (Bd) infection of H. boettgeri. It ensures that individuals are free of Bd for use in subsequent experiments.
Attachments
n4ub5hd6.docx
n4ub5hd6.docx
16KB
Guidelines
Conclusion:

This protocol outlines the steps for quarantining and conducting heat treatment on H. boettgeri frogs infected with Bd to eliminate the infection.
Materials
  • H. boettgeri adult frogs that are Bd-positive
  • 38 L glass tanks (25 × 30 × 50 cm) with plastic lids containing holes smaller than 1 cm in diameter for air circulation
  • Aquarium heater (EHEIM thermocontrol 75 W, Deizisau, Germany)
  • Aquarium filter (Tetra, Blacksburg, VA, USA)
  • EcoTestr CTS1 meter (Oakton, Vernon Hills, IL, USA)
  • Water testing kit (API Liquid Test Kits, API®, Chalfont, PA, USA)
  • Gravel substrate (diameter > 1 cm to prevent ingestion by the animals)
  • Artificial plants
  • PVC pipes
  • Reverse osmosis (RO) water
  • Synthetic sea salt mixture (Instant Ocean® Sea Salt, Blacksburg, VA, USA)
  • Sodium bicarbonate
  • Brine shrimp (Artemia sp., BIO-MARINE®, Hawthorne, CA, USA)
  • Frozen gamma-irradiated bloodworms (Chironomus sp., Tropical Marine Center, UK)
  • Powdered fry food (Sera® Micron Nature, Heinsberg, Germany)
  • Small-granule fish food (BUG BITESTM Tropical Formula, Mansfield, MA, USA)
  • Nitrifying bacterial suspension (ProLine® Aquaculture Bacteria Concentrate, Apopka, FL, USA)
  • VirkonTM Aquatic (LANXESS, Pittsburgh, PA, USA)
Quarantine setup
Quarantine setup
Place Bd-positive frogs in 38 L standard system glass tanks, condition the water, feed the animals, and conduct water changes as described in “Housing and Care for Hymenochirus boettgeri” protocol.
Note
Ensure that Bd-positive frogs are completely isolated from Bd-negative frogs to prevent further transmission.

Heat Treatment
Heat Treatment
Gradually increase the water temperature in the quarantine standard system tanks from room temperature to Temperature30 °C over a period of 10 days using an aquarium heater.

Maintain the water temperature at Temperature30 °C for Duration240:00:00 .

1w 3d
Gradually decrease the water temperature from Temperature30 °C to Temperature25 °C over a period of five days.

Monitor and maintain consistent temperature levels throughout the treatment process.
Allocation to permanent aquaria
Allocation to permanent aquaria
Monitor the animals for an additional 10 days after the heat treatment, and then transfer them to permanent aquaria.