HOW TO ANESTEZE AND SACRIFICE TEST ANIMALS Practical Objectives Students understand the reasons why a test animal is anesthetized and sacrificed, and are able to do so perform anesthesia and sacrifice test animals. Students can perform anesthesia and euthanasia on test animals that meet the requirements. Basic theory Anesthesia is a state of unconsciousness induced in animals. Anesthesia is needed especially before the animal is operated on. There are three stages of anesthesia, namely analgesia (pain relief), amnesia (memory loss) and immobilization. The drugs used to achieve anesthesia usually have different effects. Some drugs can be used individually to achieve all components of anesthesia, others can only be analgesics or sedatives and can be used individually or in combination with other drugs to achieve full anesthesia. Skeletal muscle relaxants such as Curariform or neuromuscular beta blockers (e.g. succinylcholine, decamethonium, curare, galamine, pancuronium) are not used for anesthesia and do not have an analgesic effect. They can only be used together with general anesthesia. Usually, artificial respiration is required. Physiological monitoring should also be used to assess the depth of anesthesia, where normal reflex methods will not be reliable.Experiments with animals will usually end with the animal being killed, either because its in vitro organs will be removed during or at the end of the experiment (for example observing lung histology), to assess the effects of the drug (for example the toxic effects of the drug), or because the animal is experiencing suffering or illnesses and disabilities that are impossible to recover from.The term for killing a test animal is known as euthanasia, which is a process by which an animal is killed using humanely acceptable techniques. This means the animal dies easily, quickly, peacefully with as little pain as possible. tools and materials ▪ Restraint cage ▪ Ruler ▪ Weight scales - 70% Alcohol - Ether→Chloroform - Ethyl Carbamate - Haloten - Phenobarbital Sodium Work procedures 1. Animals Used The animals used were male mice, local strains, aged between 6 – 8 weeks. 1.For euthanasia experiments use mice that meet the requirements for sacrifice, namely if an animal has lost more than 20% body weight, decreased exploration behavior, reluctance to move (decreased locomotion/mobility), bent posture, piloerection (hair standing up), moderate to severe dehydration (sunken eyes, lethargy), unrelenting pain (e.g. vocalization distress). 2.For anesthesia experiments use healthy animals 2. How to anesthetize mice Compounds that can be used for anesthesia are: Ether Ether can be used for short-term anesthesia. The ether is placed on cotton and put in a tightly closed container, then the animal is placed in the container and closed. When using ether, you should wear a mask to prevent you from inhaling the ether vapor. When the animal has lost consciousness, the animal is removed and ready for surgery. Further additions to maintain the anesthesia can be given with the help of cotton wool soaked with the drug. 3. Euthanasia of mice by physical means The physical method is done by dislocating the neck. The dislocation process is carried out by: 1.The mouse's tail is held and then placed on a surface that it can reach, allowing the mouse to stretch its body. 2.When the mouse stretches its body, a support is placed at the nape of the neck, for example a pencil or metal rod held with the left hand. 3.The tail is pulled hard with the right hand, so that the neck will be dislocated and the mouse will be killed.