Lab Animal 1st Lesson PDF

# What is One Health? The health of people is closely connected to the health of animals and our shared environment. ## Laboratory Animals: Roles in Society | **Year** | **Scientist(s)** | **Animal(s) Used** | **Contribution** | |---|---|---|---| | 1984 | Milstein, Koehler, Jerne | Mouse | Techni...

# What is One Health? The health of people is closely connected to the health of animals and our shared environment. ## Laboratory Animals: Roles in Society | **Year** | **Scientist(s)** | **Animal(s) Used** | **Contribution** | |---|---|---|---| | 1984 | Milstein, Koehler, Jerne | Mouse | Techniques of monoclonal antibody formation | | 1990 | Murray, Thomas | Dog | Organ transplant techniques | | 1997 | Prusiner | Mouse, hamster | Discovery of prions, a new biological principle of infection | | 2003 | Lauterbur, Mansfield | Clam, mouse, dog, rat, chimpanzee, pig, rabbit, frog | Discoveries concerning magnetic resonance imaging | | 2008 | Barre-Sinoussi, Montagnier | Hamster, mouse, cow | Discovery of human immunodeficiency virus | | 2008 | zur Hausen | Monkey, chimpanzee, mouse | Discovery of papilloma viruses causing cervical cancer | | 2011 | Hoffman, Beutler | Fruit fly, mouse | Discoveries concerning the activation of innate immunity | | 2011 | Steinman | Mouse | Discovery of the dendritic cell and its role in adaptive immunity | - Sources: National Association of Biomedical Research (www.nabr.org) and Nobel Prize (www.nobelprize.org). ## Animal Research Saves Animals Too _Source: Foundation for Biomedical Research_ We often do not think about the role animals have played in developing the treatments used in companion animals. From flea and tick medications, to rabies and leukemia vaccines, to treatments for diabetes and cancer, virtually every important advance and promising treatment available today to ease the pain and suffering of animals was developed with animal research. Thanks to animal research companion animals can live longer, happier, and healthier lives. ## Laboratory Animal Population _Dr. Masangkay Lecture_ A bar graph showing the number of laboratory animals from 1979 to 2000. The number of animals is on the Y axis and the year is on the X axis. ## Purposes of Experiments _Source: Commission of the European Communities. Commission Staff Working Document Annex to the Report on the Statistics on the Number of Animals used for Experimental and other Scientific Purposes in the Member States of the European Union in the year 2002 (COM(2005)7 final), Brussels._ A pie chart showing the proportion of experiments conducted for different purposes. The largest proportion is for fundamental biology studies (38.1%), followed by research and development (22.8%). ## Uses of Laboratory Animals 1 A triangle with three sections. The top section is labeled "Fundamental Research". The middle section is labeled "Applied Research". The bottom section is labeled "Education & Training". - **Fundamental research:** Increasing knowledge without immediate practical applications - **Applied research:** Practical application - **Education & Training:** Diagnostics ## Fundamental Research - Fundamental relationships between the processes which take place in the body or the causes of diseases. - Increasing knowledge without immediate practical applications - Genetically modified mouse models - How the liver deals with toxic substances ## Applied Research - **Area of science:** - The processes of life - The prevention and treatment of disease - The genetic and environmental factors related to disease and health - **Produce desired products, such as using rabbits to produce antibodies** - **Vaccine manufacture:** - Development - Potency - Safety testing "toxicity testing" ## Applied Research - **Most applied research concerns:** - Human health and human medicine - Diagnostic procedures or therapeutic treatments that may advance human health - Different animal models of human disease. - Mice are used to model cancer or diabetes mellitus. - Investigation of animal disease and production of veterinary drugs ## Diagnostic Purposes - Diagnosis of disease - About 2% - Replaced with non-animal methods ## Education and Training - Mannequins - Computer simulations - For example, normal physiology of organs such as the heart, respiratory control and kidney function can be simulated - Videos - Plasticised specimens - Ethically-sourced cadavers (obtained from animals that have been euthanased for medical reasons, or that have died naturally or in accidents) - Models - Diagrams - Self-experimentation - Supervised clinical experiences. ## Education and Training - 2% of laboratory animals - Dissections, undergraduate wet labs and research - Veterinary education or medical education - to practice surgical skills - “Hands-on" experiences are the best way to learn and that providing practical classes with animals is interesting for students - Practical skills and scientific principles ## Alternative Models in Education Six images: - A stuffed dog head - A model of a pregnant woman with the fetus visible - A model of a human head being dissected. The head is opened to show the interior of the skull. - A computer screen showing a 3D model of a tree and the words "Tree: 412P" at the top of the screen. - A person using a computer to look at a close-up image of a cell, and the words "Exploring ... cells." at the top of the screen. - A television screen showing a video of a human colon being dissected. The words "Exploring ... colon" are at the top of the screen. ## Objectives - PART 1. - LABORATORY ANIMAL ROLE IN SOCIETY - "ONE HEALTH" - PART 2. - DIFFERENT USES OF LABORATORY ANIMALS - PART 3. - WELFARE ISSUES CONCERNING LABORATORY ANIMALS - LAWS & ORGANIZATIONS GOVERNING LAB ANIMAL IN THE PHILIPPINES - PART 4. - ROLE/S OF VETERINARIANS IN LAB ANIMAL RESEARCH ## Animal Welfare Versus Animal Rights | **Animal Welfare** | **Animal Rights / Animal Protection** | |---|---| | Has a scientific definition | Has no standard definition | | Centred on science & research | Centred on philosophy and ethics | | Deals with animals' own experience of life | Condemns & restricts human use of animals | | Deals with how humans may interact with animals | Against human interactions with animals | | Focus on animal well-being in human care| Does not support animals being owned by humans | | Has measurable & objective standards | Is a belief system/ideology | | Is peer reviewed & transparent | Attempts to by-pass peer review by making changes in legislation | | Championed by Animal Welfare Scientists, Animal Carers and the Veterinary industry | Championed by extremists | | Changes in response to Social Licence to Operate | Disregards Social Licence to Operate | | Supported by ~99% of Australians | Supported by ~1.5% of Australians | | Belongs in legislation | Has no place in legislation | ## "Whenever an Animal's Life is to be Taken, It Should be Treated with the Highest Respect" _H. P. Baldos, DVM CARE_ An image of monkey sitting in a cage. ## Five Freedoms _Farm Animal Welfare Council_ - Freedom from hunger and thirst - Freedom from discomfort - Freedom from pain, injury and disease - Freedom to express normal behaviour - Freedom from fear and distress ## Welfare Issues - Sources of animals - Housing of animals - Procedures - Euthanasia ## Welfare Issue: Sources of Animals - Sources - Licensed breeding establishments - Animals bred specifically for research - Wild-caught animals - Use of stray animals from pounds or animal shelters - Stray pets from pounds is controversial - Use these 'excess' animals and thus prevent the need to sacrifice purpose-bred laboratory animals ## Welfare Issue: Sources of Animals - Controlling sources - reduce the risk of introducing disease into the animal colony. - Beneficial to overall health of the animals and any research data - Protect personnel from infection - viruses such as ebola, marburg, and herpes simian B have proved fatal to laboratory workers ## Welfare Issue: Housing of Animals - Animals spend most time in housing, not procedures - Often provided with physical needs - food, water and disease prevention - Environmental enrichment': - Provision of a stimulating environment which improves the animal's welfare An image of a mouse cage. ## Welfare Issue: Procedures - PAIN IN LABORATORY ANIMALS HISTORICALLY POORLY RECOGNISED AND TREATED - DIFFERENT SPECIES SHOW DIFFERENT BEHAVIOURS - ANIMALS ARE NOT LITTLE HUMANS - ANALGESIA AND ANAESTHESIA CAN REDUCE PAIN - PAIN SCORES CAN HELP TO MEASURE PAIN - NOT ALL PROCEDURES ARE PAINFUL - FOR EXAMPLE, MINOR PROCEDURES OR ANAESTHETIZATION - HOWEVER, SOME PAINFUL PROCEDURES MAY BE SEEN AS 'MINOR' AS THEY ARE SIMPLE OR COMMON ## Euthanasia - Euthanasia because: humane endpoint, end of trial, or integral to experiment - Ideal: - Non-painful - Time to unconsciousness and death short - Reliable and non-reversible - Minimal psychological stress to animals - Safe for operator - Death must be confirmed - Methods: injection, inhalation, physical ## Care and Use of Laboratory Animals - READING ASSIGNMENT - HIGHLY REGULATED AND - DIFFICULT RESEARCH PROJECT APPROVED - IN THE PHILIPPINES ## The Ethical Guidelines - USE OF ANIMALS IN RESEARCH ## Republic Act 8485 of 1998 - OTHERWISE KNOWN AS THE - ANIMAL WELFARE ACT - Department of Agriculture Administrative Order No. 40 series of 1998 - "Rules and Regulations on the Conduct of Scientific Procedures Using Animals," ## DA-AO no. 40 Series of 1998 - August 2, 1998 - Rules and Regulations on the Conduct of Scientific Procedures Using Animals - Signed by: DA Secretary Edgardo J. Angara - Published at: Malaya Newspaper - November 18, 1999(Thursday) - BASIS: Section 1, 2, 3, 6 of RA 8485 - Prepared by: DA-CAW and Philippine Association for Laboratory Animal Science (PALAS) ## Philippine Association for Laboratory Animal Science - Founded in 1988 - Member: AFLAS - (Asian Federation for Laboratory Animal Science Associations) - Affiliate: PVMA - (Philippine Veterinary Medical Association) - First published a COP (Code of Practice) for laboratory animals in 1993 An image of a jar with a mouse in it. ## PALAS - ESTABLISHED TO: - EMBODY THE IDEALS FOR THE DEVELOPMENT AND ADVANCEMENT OF LABORATORY ANIMAL SCIENCE - SAFEGUARD INTEREST IN MAINTAINING ETHICAL AND PROFESSIONAL STANDARDS. - IMPLEMENTATION : - ANIMAL WELFARE ACT OF 1998 - CORRESPONDING RULES AND REGULATIONS ON THE CONDUCT OF SCIENTIFIC PROCEDURES USING ANIMALS (DA-AO NO. 40) - PALAS ADVOCATES THE PRACTICE OF ANIMAL WELFARE AND GOOD SCIENCE. ## Institutional Animal Care and Use Committee (IACUC) - efforts toward common purposes: - to makes sure that the animals under their control are used and cared for humanely - The IACUC to perform its tasks properly, its members must be informed of its responsibilities. - Institutions have the responsibility to provide each IACUC members with proper **training**. - Assignment (20 points) - READ about IACUC - Guide Questions - 1. Role of IACUC in an institution - 2. Different members of IACUC - IACUC committee - 3. IACUC application form -- example ## Objectives - PART 1. - LABORATORY ANIMAL ROLE IN SOCIETY - "ONE HEALTH" - PART 2. - DIFFERENT USES OF LABORATORY ANIMALS - PART 3. - WELFARE ISSUES CONCERNING LABORATORY ANIMALS - LAWS & ORGANIZATIONS GOVERNING LAB ANIMAL IN THE PHILIPPINES - PART 4. - ROLE/S OF VETERINARIANS IN LAB ANIMAL RESEARCH ## Objectives - 1. Preclinical Drug Testing - 2. Steps of Preclinical Research - 3. Review where our profession as veterinarians or technicians would be important in the process An image of a lab rat and a bunch of test tubes. ## Preclinical Drug Testing - **Preclinical studies:** Testing of a drug, procedure or other medical treatment in animals before trials maybe given to humans A diagram showing the drug development cycle with the following stages: Drug Discovery, Pharmacology, Formulation Development, Toxicology, Phase 1, Phase 2, Phase 3, and NDA Submission. ## Drug Development Cycle - **Pharmacology stage assesses *ADME* - Absorption; Distribution; Metabolism; and Excretion.** - **Toxicology studies aim to look at the effects of longer-term drug exposure on the body** - **Investigational New Drug (IND) applications** - once approved, allow clinical trials to proceed A diagram shows the drug development cycle with the following stages: Drug Discovery, Pharmacology, Formulation Development, Toxicology, Phase 1, Phase 2, Phase 3, and NDA Submission. ## Preclinical Drug Testing A diagram showing the different aspects of preclinical drug testing. The diagram is divided into three sections: - **Body do to the drug:** Pharmacokinetics - **Drug do to the body:** Pharmacodynamics - **Drug Safety & Efficacy:** MTD/DOSE OPTIMIZATION, ADME, Tumor Inhibition, Survival Analysis, EFFICACY & MECHANISM OF ACTION, Time, Mechanism of Action, Impact on Metastasis, Gene Expression, Apoptosis/Necrosis, Function/Phenotype _Source: HHS Public Access. Author manuscript; Adv Healthc Mater. Author manuscript; published in final edited form as Adv Healthc Mater 2020 Jun 1;9(12):1901368. Best Practices for Preclinical In Vivo Testing of Cancer Nanomedicines. Danielle M. Valcourt._ ## Important Duties - Veterinarians - **Animal Welfare** - IACUC - 3R - Application - Review - Approval - Technical Training - **Generation of Preclinical Models** - Insilico - Invitro - Invivo - **Mouse Monitoring** - **Euthanasia** - **Tissue collection** - **Tissue Processing** - **Down stream Analysis** - PCR - qPCR - Western Blot - Flow Cytometry - Etc. - **Data Analysis** - **Manuscript** An image of stethoscope. ## Duties - Veterinarians - **The 3 R's of Animal Research** | **Reduce** | **Refine** | **Replace** | |---|---|---| | Reduce the number of animals used | Refine tests to cause animals less stress | Replace animal studies with other methods | - **IACUC** _Source: HHS Public Access. Author manuscript; Adv Healthe Mater. Author manuscript; published in final edited form as Adv Healthe Mater 2020 Jun 1;9(12):1901368. Best Practices for Preclinical In Vivo Testing of Cancer Nanomedicines. Danielle M. Valcourt._ ## Important Duties - Veterinarians - **IACUC** **Checklist for Establishing Animal Use Protocols** - Contact institution's IACUC and complete necessary training modules - Write Animal Use Protocol (AUP) and gain IACUC approval. Include the following components in AUP: - Non-scientific summary of scientific goals and significance of study - Detailed experimental design, including: - Species identification - Appropriateness of species and numbers used - Justification for use of animals over in vitro methods - Statistical justification for the requested number of animals - Description of proposed use of animals - Detailed description of planned procedures - Description of criteria and methods for euthanasia **Checklist for Maintaining Animal Welfare Throughout Protocols** - Perform studies, consulting with veterinary staff as needed to ensure animal welfare. Throughout experiments, monitor animals for: - Tumor size (maintain diameter below 1.2 cm for subcutaneous tumors in mice) - Weight (ensure weight loss does not exceed 20%) - Aggressive behavior (separate animals into distinct cages if needed) - Signs of distress (hunched posture, difficulty ambulating, ruffled fur, labored breathing, abdominal distension, persistent hypothermia, incontinence or diarrhea, discharge of blood, mucus, or pus) ## Types of Genetically Engineered Mice A diagram with two panels. The panels are labeled "A" and "B". **Panel A** - Transgenic mice - Add randomly - Knock in - Add specific site - Knockout - Eliminate gene global/whole - Conditional Eliminate gene (specific site) - increase specificity of genetic targeting A diagram showing a gene with a floxed allele, a WT allele, a WT allele that has been activated, and a Gapdh allele. **Panel B** - A murine model of chronic lymphocytic leukemia based on B cell-restricted expression of Sf3b1 mutation and Atm deletion - Shanye Yin, Rutendo G. Gambe, Jing Sun, Aina Zurita Martinez, Zachary J. Cartun, Fara Faye D. Regis, Youzhong Wan, Jean Fan, Angela N. Brooks, Sarah E.M. Herman, Elisa ten Hacken, Amaro Taylor-Weiner, Laura Z. Rassenti, Emanuela M. Ghia, Thomas J. Kipps, Esther A. Obeng, Carrie L. Cibulskis, Donna Neuberg, Dean R. Campagna, Mark D. Fleming, Benjamin L. Ebert, Adrian Wiestner, Ignaty Leshchiner, James A. DeCaprio, Gad Getz, Robin Reed, Ruben D. Carrasco, Catherine J. Wu, and Lili Wang A diagram showing two lanes of electrophoretic gel. Each lane is labeled "Sf3b1" and shows different PCR bands from either B cells or T cells. ## Mouse Model Generation : Recommendation - Breeding - Goal - Quantity of mice - Genotype - Future experiments - You need to choose the right breeding pair - Right genotype - Good breeders - Genotype properly - PCR - Primers - Reaction - Identify mice - Weaning Record - Monitoring of mice - General condition - Blood An image of a mouse. ## Review - Preclinical studies - Testing of a drug, procedure or other medical treatment in animals before trials maybe given to humans - Preclinical Pharmacology - ADME - Absorption; Distribution; Metabolism; and Excretion - Preclinical Toxicology - toxicity of the drugs - aim to monitor for any adverse effects that could arise as a result of taking the drug - Veterinarian - Ensure that research is conducted responsibly and competently - Animal Welfare - Provide knowledge and skills required to design and carry out experiment - 3Rs - 3Rs - Reduce the number of animal used - Refine the procedures you will do to the animal - Replace invivo testing with different models example invitro or insilico.