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Ethics Statement

At Quantum Biology Tech (QuBiT) Lab, we are committed to upholding the highest scientific and ethical standards in all aspects of our research endeavors.

 

Our dedication to the principles known as “the 3 R’s” – Replacement, Reduction, and Refinement – guides our efforts to replace animal models with alternative methods, reduce the number of animals used in research, and refine experimental procedures to minimize animal suffering.

All of our researchers undergo training, and our research protocols undergo thorough review and approval by our Institutional Animal Care and Use Committee (IACUC), ensuring compliance with federal regulations, industry best practices, and institutional policies regarding animal welfare. The inclusion of a veterinarian on our IACUC ensures the highest standards of veterinary care and husbandry practices for our animal subjects.

In line with our commitment to transparency and accountability, we proudly support open science initiatives and will be broadcasting our study live, providing the public with unprecedented access to the scientific process. This initiative underscores our belief in the importance of public engagement and scientific outreach, as well as our willingness to go above and beyond what is required of us to promote responsible research practices.

QuBiT Lab Protocols for Humane Euthanasia of Laboratory Animals

Introduction

The QuBiT Lab acknowledges our responsibility to ensure the well-being of the animals we use in research, and that they are treated humanely (1). This policy provides protocols for the humane euthanasia of lab animals, by:

  1. Following laws and guidelines for euthanasia established by governing institutions as well as best practices in the professional community;

  2. Establishing procedures that make the process as fast, painless, and distress free as possible for the animal (2);

  3. Establishing procedures that minimize psychological stress on personnel (3).

Training of personnel

The process of euthanasia should only be conducted by trained personnel (4), enabling accurate identification of lab animals that should be euthanized, as well as the correct administration of euthanasia in a way that is pain and stress free for both lab animals and personnel. Training include:

  1. General information about working with the specific animal;

  2. Conducting euthanasia.

The researchers all completed the online AALAS “Laboratory Animal Research Support Certificate”. Proof of completion of training, such as certificates, is to be reviewed by the IACUC and veterinarian.

Well-being of personnel

A notice is posted in the lab, addressing psychological stress that can develop in lab personnel who observe or conduct euthanasia. It includes:

  1. The probability that psychological stress can develop in lab personnel who observe or conduct euthanasia (3);

  2. That researchers who voice concern about stress in themselves or others will not be punished;

  3. Indicators of psychological stress in lab personnel, stemming from conducting or observing euthanasia of lab animals (3);

  4. That steps are being taken to minimize likelihood of stress developing in lab personnel (3), as per the below sections: Use of a Daily Welfare Assessment and Use of a Euthanasia Log.

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Use of a Daily Welfare Assessment

A welfare assessment of the animals should be conducted daily, to determine if any should be euthanized (5).

The assessment or rubric used is to be approved by the IACUC (6). It includes:

  1. Examples of behaviors or physical features that indicate the animal is likely experiencing pain, distress, or loss of normal function;

  2. A point system with a designated value for the severity or negative impact on well being, of each indicator;

  3. A clear description of when the animal should be euthanized, for example if the animal is displaying a total of 3 points from all indicators.

Use of a Euthanasia Log

Use of euthanasia log of which lab personnel have administered euthanasia will aid in ensuring the responsibility is spread between lab personnel. It includes:

  1. Date and time;

  2. Who administered euthanasia;

  3. How many animals were euthanized;

  4. Which method was used

Posting of a Material Safety Data Sheets (MSDS) for each chemical used in the euthanasia process

In the lab, there is a posted MSDS for all chemicals used in the euthanasia process, including:

  1. Appropriate PPE required;

  2. Known dangers of the chemical;

  3. First aid depending on type of exposure.

QuBiT Lab Protocols for hormone injection

Injection of hormone to stimulate egg maturation and laying in mature females and stimulate mating behavior in mature males.  This involves a single injection or serial priming and boosting injections separated by one or more days of human chorionic gonadotropin (HCG) into the dorsal lymph sac (Dosages:  500 IU for X laevis, and 200 IU for X tropicalis). Pregnant mare serum gonadotropin (PMSG) is also used for priming at a dose of 50U/frog for X. laevis and 20U/frog for X. tropicalis. We also use Luteinizing Hormone (LH) to induce ovulation. LH is an alternative to HCG, which we have found works efficiently in Xenopus at a dose of 2.5ug/g for X. laevis and 5ug/g for other species. This amounts to 150-200ug for X. laevis and 50-100ug for X. tropicalis and other species. A second injection of hormone (superovulation) may be used a week later for those frogs that do not lay. For in vitro fertilization, male frogs are euthanized as described below and the testes isolated. A piece of testes is removed and macerated and placed on eggs.

 

All injectable pharmaceuticals are purchased at the ultrapure level and purity levels are included and assured by the manufacturer.  Working concentrations are diluted to proper dosage using sterile RO water or PBS.  If appropriate, working solutions are microfiltered (0.2 µm) before use.

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Resources

  1. Guide for the Care and Use of Laboratory Animals Eighth Edition (pg 1). https://grants.nih.gov/grants/olaw/guide-for-the-care-and-use-of-laboratory-animals.pdf

  2. American Veterinary Medical Association Guidelines for the Euthanasia of Animals, 2020 Edition (pg 6-8). https://www.avma.org/sites/default/files/2020-02/Guidelines-on-Euthanasia-2020.pdf

  3. American Veterinary Medical Association Guidelines for the Euthanasia of Animals, 2020 Edition (pg 15). https://www.avma.org/sites/default/files/2020-02/Guidelines-on-Euthanasia-2020.pdf

  4. American Veterinary Medical Association Guidelines for the Euthanasia of Animals, 2020 Edition (pg 9). https://www.avma.org/sites/default/files/2020-02/Guidelines-on-Euthanasia-2020.pdf

  5. Canadian Council on Animal Care Guidelines, Amphibians (pg 57. https://ccac.ca/Documents/Standards/Guidelines/CCAC_Guidelines-Amphibians.pdf

  6. Guide for the Care and Use of Laboratory Animals Eighth Edition (pg 27-28). https://grants.nih.gov/grants/olaw/guide-for-the-care-and-use-of-laboratory-animals.pdf

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