Alternate animal experiments ( Basic Introduction )
- 1. ALTERNATE ANIMALALTERNATE ANIMAL EXPERIMENTSEXPERIMENTS Presented by :-Presented by :- Deepak Chandra JoshiDeepak Chandra Joshi M.Pharma (Pharmacology)M.Pharma (Pharmacology) Department of Pharmaceutical SciencesDepartment of Pharmaceutical Sciences Bhimtal (Nainital)Bhimtal (Nainital)
- 2. Table of Content :-Table of Content :- IntroductionIntroduction Need for alternative to animalNeed for alternative to animal Laws and regulationsLaws and regulations Alternative experiments methodsAlternative experiments methods 1.1. RefinementRefinement 2.2. ReductionReduction 3.3. ReplacementReplacement In-Vitro MethodsIn-Vitro Methods
- 3. IntroductionIntroduction Alternative animal experiment are the develepmAlternative animal experiment are the develepm -ent and Implementation of test method’s that-ent and Implementation of test method’s that avoid the use of life animals.avoid the use of life animals. The primary goal of alternative animal experimeThe primary goal of alternative animal experime -nt is the minimize pain which is suffer by-nt is the minimize pain which is suffer by animal during the experiments.animal during the experiments.
- 4. Animal are used in Science for:-Animal are used in Science for:- UG teaching to learn physiological mechanismUG teaching to learn physiological mechanism ,anatomy and effect of various drugs on human,anatomy and effect of various drugs on human body.body. PG teaching to show effects of various drugs toPG teaching to show effects of various drugs to find out the nature of unknown drug and it’sfind out the nature of unknown drug and it’s effect or for bioassayeffect or for bioassay In research to understand the working of body atIn research to understand the working of body at disease and healthy statedisease and healthy state In research to conduct screening for drugsIn research to conduct screening for drugs bioassay and pre clinical testing of new drugsbioassay and pre clinical testing of new drugs
- 5. Animals are used to test possibilities that would beAnimals are used to test possibilities that would be difficult or impossible to test using the target speciesdifficult or impossible to test using the target species (humans).(humans). It is mandatory to do extensive toxicological studies inIt is mandatory to do extensive toxicological studies in animals before the drug gets approval for clinical trialsanimals before the drug gets approval for clinical trials in humans.in humans. ““There is no doubt that the best test species for humansThere is no doubt that the best test species for humans are humans. It's not possible to extrapolated animal dataare humans. It's not possible to extrapolated animal data directly to humans you to inter species variation indirectly to humans you to inter species variation in anatomy ,physiology and biochemistry”.anatomy ,physiology and biochemistry”.
- 6. Needs for Alternatives :-Needs for Alternatives :- In Laboratory an animal may be….In Laboratory an animal may be…. 1. Poisoned.1. Poisoned. 2. Deprived of food ,water and sleep.2. Deprived of food ,water and sleep. 3. Applied with skin and eye irritants.3. Applied with skin and eye irritants. 4. Subjected to physiological stress.4. Subjected to physiological stress. 5. Deliberately infected with disease.5. Deliberately infected with disease. 6. Brain damaged, paralysis, surgical mutilated.6. Brain damaged, paralysis, surgical mutilated.
- 7. Disadvantages of animal experiments :-Disadvantages of animal experiments :- 1.1. Pain, distress and unethical behavior toPain, distress and unethical behavior to animals.animals. 2.2. Requirement of skilled manpower.Requirement of skilled manpower. 3.3. Time consuming protocol.Time consuming protocol. 4.4. High Cost.High Cost.
- 8. Laws and Regulations:-Laws and Regulations:- YEARYEAR LAWLAW 19601960 Prevention of cruelty to animal act. (amend.1982).Prevention of cruelty to animal act. (amend.1982). 19641964 Committee for the purpose of control andCommittee for the purpose of control and supervision of experimental animal (CPCSEA).supervision of experimental animal (CPCSEA). 19921992 Indian national science academy. “Guidelines forIndian national science academy. “Guidelines for care and use of animals in scientific research.care and use of animals in scientific research. 20012001 Indian Council of medical Research (ICMR).Indian Council of medical Research (ICMR). ““Guidelines for use of laboratory animals inGuidelines for use of laboratory animals in medical collages”.medical collages”. 20092009 MCI amendment to use alternatives to replaceMCI amendment to use alternatives to replace animal experiments.animal experiments. 20132013 UGC guidelines for discontinue of animalUGC guidelines for discontinue of animal experiments in zoology or life science in a phasedexperiments in zoology or life science in a phased
- 9. Modified use of animals:-Modified use of animals:- Russel and Burch in 1959 proposed that ….Russel and Burch in 1959 proposed that …. ““If animals were to be used in experiments every effortIf animals were to be used in experiments every effort should be made to replace them with non-sentientshould be made to replace them with non-sentient alternatives”.alternatives”. They developed the 3R strategy which includes:-They developed the 3R strategy which includes:- 1.1. Refinement:-Refinement:- refine experimental methods to decreaserefine experimental methods to decrease unnecessary pain and trauma to animals.unnecessary pain and trauma to animals. 2.2. Reduction:-Reduction:- reduce the no. of animals used in thesereduce the no. of animals used in these experiments.experiments. 3.3. Replacement:-Replacement:- replace the animal experiments .replace the animal experiments . ex.Computar stimulation model , in-vitro method , cellex.Computar stimulation model , in-vitro method , cell culture technique.culture technique.
- 10. 1.Refinement1.Refinement Setting the earliest possible end point.Setting the earliest possible end point. Using appropriate analgesics and anesthetics forUsing appropriate analgesics and anesthetics for painful procedure.painful procedure. Use proper handling technique for animals.Use proper handling technique for animals. Adequate training prior to performingAdequate training prior to performing experiment.experiment. Ensure drug dose are correct and srug are notEnsure drug dose are correct and srug are not expired.expired. Performed surgeries and procedure aseptically toPerformed surgeries and procedure aseptically to prevent infection.prevent infection.
- 11. 2.2.ReductionReduction Perform pilot studies.Perform pilot studies. Design studies to use animals as their ownDesign studies to use animals as their own control. ex. Cross over study.control. ex. Cross over study. Gather data for more than one experimentGather data for more than one experiment concurrently.concurrently. Consult with statistician and use minimum no. ofConsult with statistician and use minimum no. of animals.animals. Minimize variables such as disease , diet , stress,Minimize variables such as disease , diet , stress, genetics.genetics. Use appropriate species of animals.Use appropriate species of animals.
- 12. 3.Replacement3.Replacement Substitution of insentient material in place ofSubstitution of insentient material in place of conscious higher animals.conscious higher animals. Could be relative or absolute.Could be relative or absolute. Replace higher animals with lower animals .Replace higher animals with lower animals . Replace live animals with dummies for teachingReplace live animals with dummies for teaching and dissection purpose .and dissection purpose . Use computer simulation and in-vitro methods .Use computer simulation and in-vitro methods . Use cell culture and tissue culture.Use cell culture and tissue culture.
- 13. 1. In-Vitro Models:-1. In-Vitro Models:- In-Vitro biomedical research entails theIn-Vitro biomedical research entails the maintenance of organs, tissues (fragments ofmaintenance of organs, tissues (fragments of organs and tissues), and cells outside the body.organs and tissues), and cells outside the body. Can be grown as independent cell lines orCan be grown as independent cell lines or preserve the architecture of the entire organ aspreserve the architecture of the entire organ as organ culture and tissue culture.organ culture and tissue culture. Stem cells are also used in in-vitro models.Stem cells are also used in in-vitro models.
- 14. Source of tissue for in-vitro methods:-Source of tissue for in-vitro methods:- Avian – Chick embryosAvian – Chick embryos Rodents – rats and mice (wild types and adult traRodents – rats and mice (wild types and adult tra -nsgenic). Embryonic, post-natal and-nsgenic). Embryonic, post-natal and and adult.and adult. Human – 1. Neural progenitor cells from abortedHuman – 1. Neural progenitor cells from aborted fetuses and stem cell line.fetuses and stem cell line. 2. cord blood derived stem cells.2. cord blood derived stem cells.
- 15. Types of in-vitro System:-Types of in-vitro System:- Cell Culture Cell Lines Primary culture Organ architecture preserved
- 16. In-vitro methods :-In-vitro methods :- In-vitro pyrogen test .In-vitro pyrogen test . Embryonic stem cell test .Embryonic stem cell test . Local lymph node assay for skin sensitization.Local lymph node assay for skin sensitization. Clinical skin patch test on human volunteers.Clinical skin patch test on human volunteers. Neutral red uptake assay.Neutral red uptake assay. Carcinogenicity test.Carcinogenicity test. Acute toxicity test.Acute toxicity test. Repeated dose toxicity test.Repeated dose toxicity test. Development neurotoxicity test.Development neurotoxicity test.
- 17. 2.Microorganism based model:-2.Microorganism based model:- Tetrahymena pyriformis – a ciliate protozoanTetrahymena pyriformis – a ciliate protozoan being used to study the effects of anesthetics onbeing used to study the effects of anesthetics on metabolism.metabolism. Salmonella typhimurium – bacteria used inSalmonella typhimurium – bacteria used in mechanistic studies in genetics as well as themechanistic studies in genetics as well as the Ames mutagenicity test.Ames mutagenicity test.
- 18. 3.3.In-chemico testing :-In-chemico testing :- The toxic potential of substances can sometimes beThe toxic potential of substances can sometimes be detected using relatively simple chemistry baseddetected using relatively simple chemistry based methods and not requiring human cells.methods and not requiring human cells. ex. HPLCex. HPLC Direct peptide relativity assay – used to assess whether aDirect peptide relativity assay – used to assess whether a chemical or cosmetics will cause allergy.chemical or cosmetics will cause allergy. The test works by mimicking a key step in theThe test works by mimicking a key step in the development of allergies – the binding of proteinsdevelopment of allergies – the binding of proteins found in the skin to substance.found in the skin to substance. If proteins bind to the substance them its very unlikelyIf proteins bind to the substance them its very unlikely that it will cause an allergic reaction.that it will cause an allergic reaction.
- 19. 4.4.In-silico Models :-In-silico Models :- Computer aided molecular drug design.Computer aided molecular drug design. Quantitative structure activity relationships.Quantitative structure activity relationships. Computer assisted learning.Computer assisted learning. Computer or mathematical analysis.Computer or mathematical analysis. Microfludic chipsMicrofludic chips DNA chips.DNA chips. Organ on chipOrgan on chip Human on chipHuman on chip
- 20. 5.5.Computer assisted learning (CAL) :-Computer assisted learning (CAL) :- CAL deals with a range of software packagesCAL deals with a range of software packages which simulate the animal experiments.which simulate the animal experiments. Two software's are currently used in india.Two software's are currently used in india. 1.1. Ephram – developed by JIPMER, India.Ephram – developed by JIPMER, India. 2.2. X-cologyX-cology
- 21. ExpharmExpharm Contains programs on……..Contains programs on…….. 1.1. Effects of drugs on rabbit eye.Effects of drugs on rabbit eye. 2.2. Bio assay of histamine using guinea pig ileum.Bio assay of histamine using guinea pig ileum. 3.3. Effects of drug on frog heart.Effects of drug on frog heart. 4.4. Effects of drugs on dog blood pressure and HREffects of drugs on dog blood pressure and HR 5.5. Effects of drugs on cillary movement of frogEffects of drugs on cillary movement of frog esophagus.esophagus. The user can conduct experiment and collect data.The user can conduct experiment and collect data. Each program can be run in two modes :-Each program can be run in two modes :- 1.1. Tutorial modeTutorial mode 2.2. Examination modeExamination mode
- 22. THE ENDTHE END DEEPAK JOSHIDEEPAK JOSHI M.PHARMAM.PHARMA (PHARMACOLOGY)(PHARMACOLOGY)