Mice euthanasia methods. The objective of our study was to assess When CO 2 is effective as a primary euthanasia method and when other primary methods should be utilized. Although striving for a science-based report in which “only those methods for which reliable information could be obtained were included,” 9 the Panel nonetheless approved, without citation of supporting data, “disarticulation of the skull and cervical vertebrae” as a method of mouse Assessing Cervical Dislocation as a Humane Euthanasia Method in Mice. Non-weaned rodents between 10-21 days old. J Am Methods of Euthanasia Carbon dioxide • Considered an acceptable agent of euthanasia for laboratory rats, mice, and guinea pigs with the following conditions. Euthanasia methods included: halothane inhalation, carbon dioxide inhalation, intraperitoneal sodium pentobarbital, intravenous potassium chloride, and cervical dislocation with and without inhalation is considered an acceptable method of euthanasia for small rodents (e. The IAS Literature searches for alternative methods of humane euthanasia are required as part of the IACUC protocol. 1. Deposit scholarly works such as posters, presentations, research protocols, conference papers or white papers. 2. The Animal Welfare Act (AWA) and Animal Welfare Regulations (AWR), 9 CFR Part 1, §1. Boivin GP, Bottomley MA, Schiml PA, et al. These Postmortem changes (PMCs) not only affect the results of experimental studies, but also determine postmortem intervals in forensic sciences. We evaluated the voluntary oral administration of a euthanasia drug in 99 male and 81 female mice of various strains. Rutgers Animal Care, Rutgers University, Piscataway, New Jersey. Jumoke Fasoyinu, Standards Coordinator and Copy Editor Simple Summary. Compressed CO 2 gas in cylinders is the only recommended source of carbon dioxide for euthanasia. an d the When CO 2 is effective as a primary euthanasia method and when other primary methods should be utilized. The Institutional Animal Care and Use Committee (IACUC) recommends the following euthanasia methods for neonatal rodents: • Overdose of injectable anesthetic, given IP. The Canadian Council for the Care of Laboratory Animals in its guidelines Mice and Rats . In the medical field, it is often defined as the act of painlessly killing an individual who is suffering from an incurable or painful disease. ) Reference . 4. Effects of Different Grades of Carbon Dioxide on Euthanasia of Mice (Mus musculus). We placed the cookie In conclusion, we showed that mice euthanized with a CO 2 overdose have significantly different retinal dopamine levels compared to mice euthanized with cervical dislocation. The mouse is the most commonly used model in biomedical research, particularly for fundamental studies on gametes and embryos. Intraperitoneal pentobarbital is acceptable for smaller mammals when intravenous access is not practical. Julie Dale, Standards Development Director (Project Lead) Ms. Poster, Presentation, Protocol or Paper. 4 Methods of Humane Euthanasia in Mice Page 1 30/05/2019 . Mouse acclimation to a light–dark apparatus was used to create a light aversion test based on an unconditioned preference for dark versus Refinement of experimental procedures in animal research has the objective of preventing and minimizing pain/distress in animals, including the euthanasia period. Acceptable methods of euthanasia of adult animals Laboratory animals Animal Type Recommended Acceptable Mice and rats •Overdose of inhalational anaesthetic • Carbon dioxide inhalation Commonly Used Anesthesia/ Euthanasia Methods for Brain Collection Differentially Impact MAPK Activity in Male and Female C57BL/6 Mice 2 for euthanasia of rodents. the existing evidence regarding the biological interference of the anesthetics when employed in laboratory rat and mice euthanasia Using this gradual fill method, CO 2 flow should be maintained for at least 1 minute after respiratory arrest for adult rodents and a confirmatory method of euthanasia appropriate to the species must be used. 5 mL of 70% ethanol induced death in 2 min 41 s (± 52 s) and that “no discomfort was observable in these mice. Overview of killing methods from published euthanasia guidelines for adult laboratory rodents across scientifically advanced countries. 5 min in carbon dioxide The CO2 method for primary euthanasia is effective in: Weaned rodents. 1 Subject: Humane Killing Method for Rodents via Barbiturate or Injectable Anaesthetic Date Issued: August 21, 2012 Revision: 3. Species. All currently accepted methods of euthanasia for laboratory mice involve some degree of stress, fear, anxiety, or pain. We have assessed the time to death of foetuses after performing either indirect (dam euthanasia) or direct (via intraplacental injection – a new approach to euthanasia) euthanasia methods in order to determine a euthanasia method that is appropriate, ethical and efficient for the killing of mouse foetuses. Although striving for a science-based report in which “only those methods for which reliable information could be obtained were included,” 9 the Panel nonetheless approved, without citation of supporting data, “disarticulation of the skull and cervical vertebrae” as a method of mouse euthanasia. Chick behaviors, including ataxia, loss of posture, convulsions, cessation of vocalization, and To reliably effect euthanasia in mice and rats the dose of pentobarbitone must be >200mg/kg. Neonatal mice (n = 76; age, 1 or 2 d) were exposed to isoflurane in a sealed, quart-size (0. Cervical dislocation (not acceptable for rats > 200 grams of body weight or hamsters due to Whereas fetuses at this age are resistant to inhalant anesthetics, such as isoflurane or CO 2, euthanasia may be induced by the skillful injection of chemical anesthetics or by a physical Guidelines: Euthanasia of Mice and Rats. Objective: To provide guidance on humanely euthanizing mice and rats . CO 2 administration euthanasia method had an in-between foetal time value to death. Three common approaches are carbon dioxide (CO 2) inhalation, pentobarbital injection and decapitation. 1 Small Mouse Cage, Thoren Caging Systems). Potential areas for fur As per the UACC policy on humane killing of animals used in science, overdose of barbiturates or injectable anaesthetic is an acceptable method of humane killing for rodents when conducted properly. • All animal euthanasia must be performed by appropriately trained personnel (Guide and AVMA), listed Death to be verified afterwards or ensured by physical method. Our results indicate that the most common method of euthanasia for laboratory mice – exposure to CO 2 – is at least moderately aversive to mice. As for the direct euthanasia method assessed, foetal time to death after intraplacental injection of sodium pentobarbital . Avian Wildlife Euthanasia Techniques Renée Schott, DVM, CWR LafeberVet Non-Interactive Webinar as long as 50 min to achieve euthanasia. This presentation uses cases to discuss practical euthanasia methods for reptiles and the physiology behind these methods. The principal Carbon dioxide (CO2) is one of the most commonly used gas euthanasia agents in mice, despite reports of aversion and nociception. CD of mice holds the potential for unsuccessful euthanasia, that anesthesia could be valuable for CD skills training and assessment, and that postmortem radiography has minimal promise in quality-control assessments. A secondary method of death must be employed following CO2 euthanasia. The PHS Policy, IV. sc. S. J Am Assoc Lab Anim Sci 2017; 56: 69–78. A. Conlee and Martin Request PDF | Review of Rodent Euthanasia Methods | The optimal choice of euthanasia method for laboratory rodents depends on a number of factors, including the scientific goals of the study, the The effects of alternative euthanasia methods, including carbon dioxide (CO2) or nitrogen (N2) inhalation, and a commercial negative pressure stunning system on behavioral and physiological responses of day-of-hatch male layer chicks, were evaluated in a field trial. 4 kb) Authors: Schoell, Adam R 1; Heyde, Bruce R 2; Weir, Dana E 1; Chiang, Po-Chang 3; Hu, Yiding 2; Tung, David K 3; Carbon dioxide (CO2) is one of the most commonly used gas euthanasia agents in mice, despite reports of aversion and nociception. Document No: 10. These investigators also Euthanasia Method in Mice Larry Carbone,1,* Elizabeth T Carbone, 1 Elizabeth M Yi, Diana B Bauer, Krista A Lindstrom, 1 John M Parker, Jamie A Austin,1 Youngho Seo,2 Anisha D Gandhi,3 and James D Wilkerson1 Research investigators often choose to euthanize mice by cervical dislocation (CD) when other methods would interfere with the aims of a research project. The AVMA (American Veterinary Medical Association) Panel on Euthanasia recommends the use of CO 2 with 10% to 30% exchange of chamber gas [19]. decapitation, cervical dislocation, exsanguination or removal of vital organs) be employed prior to carcass disposal in ALL rodent species. 3: Adjunctive methods (second step) include 70% alcohol and neutral-buffered 3. Methods Eight-week Rodent Euthanasia | Neonatal Rodent Euthanasia | Rabbit Euthanasia | Amphibian, Fish & Reptile Euthanasia | Compassion Fatigue Euthanasia Euthanasia is the act of killing animals using methods that cause minimal animal pain, distress and anxiety prior to rapid loss of consciousness and death. Acceptable Methods with Conditions b. Decapitation with guillotine only after the animal has been sedated (unless otherwise justified to U. Inhalation of carbon dioxide (CO 2), delivered using a gradual fill protocol, is a common euthanasia method for laboratory mice [1,2,3]. Chemical euthanasia with an overdose of intravenous pentobarbital - based euthanasia solution is the preferred euthanasia method for mammals with practical vascular access (e. e. Keywords: Euthanasia, mouse, isoflurane, cervical dislocation, oocyte Laboratory Animals 2012; 46: 167– 169. For an anesthetic method to be considered acceptable, it would ideally minimize current methods of mouse euthanasia: refinements and aversion by carly moody b. Therefore, CO 2 must not be used in University of Washington protocols for the euthanasia of animals such as rabbits, cats, swine, dogs, or nonhuman primates. Within the scope of refinement, our aim was to investigate if isoflurane and sevoflurane are an appropriate alternative killing method to CO2 in mice. Methods such as exsanguination, bilateral thoracotomy, vital organ harvest, decapitation or cervical dislocation are acceptable. Here we compared behavioural and electrophysiological reactions to CO2 or N2 at either slow fill or rapid fill in C57Bl/6 mice undergoing gas This study aimed to evaluate the pain associated with six methods of euthanasia: pentobarbital, CO 2, decapitation, isoflurane, ketamine + xylazine, and ketamine + CO 2 in Wistar rats, by applying the Rat Grimace Scale (RGS), comparing the scores, and determining the method with the highest score that might indicate pain in laboratory rodents. This study aimed to evaluate the pain associated with six methods of euthanasia: pentobarbital, CO 2, decapitation, isoflurane, ketamine + xylazine, and ketamine + CO 2 in Wistar rats, by applying the Rat Grimace Scale (RGS), comparing the scores, and determining the method with the highest score that might indicate pain in laboratory rodents. This study compared mouse aversion to isoflurane versus gradual-fill CO 2 gas, and compared two methods of isoflurane delivery: vaporizer and drop. To date, only a few publications cite this method, 31–33 and it was experimentally evaluated as a method of euthanasia in only one of these. When properly performed by competent persons using appropriate techniques, euthanasia is humane. However, studies suggest that a considerable number of researches are based on improper use or reporting of A&A methods [1, 2]. A. RATS Drug(s) or Method Dose (mg/kg) Route Anticholinergics Atropine 0. 13–15 The use of euthanasia throughout guidance and regulation protecting more efficient method for euthanasia in small rodents in animal house facilities must still be identified [3,14]. 05 SC,IM,IV Glycopyrrolate 0. Pregnant rodents. Introduction: Carbon dioxide (CO2) inhalation is a common method of euthanasia used at NIH for small rodents (e. Use CO2 to asphyxiate the rodent. This work is designed to evaluate the consequence of this method of sacrifice on the cerebellum and Euthanasia is an acceptable method for relieving pain or distress that cannot be controlled by other means. Although striving for a science-based report in which “only those methods for which reliable information could be obtained were included,”9 the Panel nonetheless approved, without citation of supporting data, “disarticulation of the skull and cervical vertebrae” as a method of mouse euthanasia. Animal Research Committee (ARC) Euthanasia of Adult Rodents by Physical Methods. Resistance to hypoxia results in a prolonged time to unconsciousness when CO 2 inhalation is used as a euthanasia agent. Methods for the euthanasia of neonatal mice includes scissor decapitation or injectable anesthetic followed by a physical means of euthanasia. Multiple groups of mice were exposed to 100% CO2 for time period Consideration must be taken about the place to carry out euthanasia: preferably in a separate room so that if distress vocalizations or the secretion of pheromones are elicited, they cannot be heard or smelled by other animals in the area (AVMA, 2020). 14 1. Although TTD did not differ between ethanol and pentobarbital-phenytoin at 28 d of age, the TTD in 3 of 12 mice was longer than 15 min after ethanol administration at this age, therefore, ethanol should not be used as a method of euthanasia for mice younger than 35 d, because the criteria for humane euthanasia were met only in mice 35 d or older. Laboratory rodents are commonly euthanized by exposure to gradually increasing concentrations of carbon dioxide (CO 2). Much of the discussion about the humane killing of research animals centres on rodents. Secondary methods: Among the indirect euthanasia methods assessed, the administration of a sodium pentobarbital overdose to pregnant female mice was found to be the fastest for foetuses, with an average post-treatment foetal death of approximately 29. • Cervical dislocation may be used to euthanize poultry of appropriate size and species when performed by a competent individual. Isoflurane and carbon dioxide (CO 2) gas are used for rodent euthanasia. It is not realistic or appropriate to inject ~50mg/kg and expect this dose to kill the animal. . Overdose of sodium pentobarbital (PB) by intraperitoneal (IP) injection is a widely accepted technique in laboratory rats, but questions remain regarding pain associated with administration. Here we compared behavioural and electrophysiological reactions to CO2 or N2 at either slow fill or rapid fill in C57Bl/6 mice undergoing gas of individual performing the euthanasia to prevent welfare concerns. Mice show aversion to CO 2 concentrations as low as 13. The UACC requires that a Primary (rendering death) and Secondary (confirmation of death) method of human killing be Species: Mice or Rats . a. 95-L) plastic bag at room temperature. 6. Cervical Dislocation • Cervical dislocation may be used to euthanize small birds, poultry, mice, immature rats (<200 g) and rabbits. NOTE: LAB_012 Euthanasia - Transcardial Perfusion in Mice and Rats, identifies that pentobarbitone may be administer ed at a dose of >50mg/kg. C. • All methods of euthanasia must be detailed on the IACUC-approved protocol to which the animal is assigned. Affiliations. Several recent publications provide clarity on the use of CO2 in laboratory rats and mice. p. Document No: 7. 6 Anesthesia in Mice). 19 Since then, with no studies of CD in In light of these results, we conclude that cervical dislocation is the best method of mouse euthanasia for obtaining intact oocytes for biomedical research. 7. Lee Niel, University of Toronto Dr. This study was designed to assess the effectiveness of carbon monoxide (CO) as an alternative euthanasia agent for mouse and rat pups on postnatal days (PND The mice brains were collected in a separate suite at the same time of the day during their active cycle following four different anesthesia and euthanasia methods: (1) Ketamine/Xylazine: mice were decapitated 45 min following an intraperitoneal injection of 100 mg/kg (#VINB-KET0-7021, Henry Schein Animal Health, Dublin, OH, USA) with 10 mg/kg The mouse is the most commonly used model in biomedical research, particularly for fundamental studies on gametes and embryos. Approved, humane methods of euthanasia are designed to minimize the potential for pain or discomfort, but may also influence the measurement of experimental variables. Rodent euthanasia methods under scrutiny 2012-Dec-19. Physiologic, behavioral, and histologic responses to various euthanasia methods in c57bl/6ntac male mice. }, author={Larry Carbone and Elizabeth T Carbone and Elizabeth M Yi and Diana Bauer and Krista A Lindstrom and John M Parker and Jamie Austin and University Animal Care Committee Standard Operating Procedure. Once the mice were unconscious (at approximately DOI: 10. Carbon dioxide (CO 2) is a widely used method of euthanasia for rodents in research settings, and is one of the most common euthanasia methods used for mice. 19 7 Anesthesia and analgesia (A&A) are the main components of many procedures on laboratory animals. A combination of inhalation (primary) followed by a physical (secondary) method is recommended for all rodents as they, especially neonates, resist inhaled agents euthanasia. The use of carbon dioxide for the euthanasia of adult rodents has become a subject of considerable debate and scientific study. 19 The mouse is the most commonly used model in biomedical research, particularly for fundamental studies on gametes and embryos. Inert gases such as nitrogen (N2) may be a viable alternative to carbon dioxide. 3. Wendy Clarence, Research Analyst Ms. The AVMA Guidelines for the Euthanasia of Animals: 2020 Edition 1 describes that certain critical factors must be 6. Schoell et al (2009). The euthanasia methods that induced the most investigate the possible protective effects of Thymoquinone against liver injury in hyperlipidemia-induced LDL-R−/− mice. 116. Location: Queen’s University Responsibility: Principal Investigators (PI), Research Staff, Veterinary Staff Purpose: The purpose of this Standard Operating Procedure (SOP) is to This SOP provides guidelines for the humane euthanasia of rodents using several techniques. Though this is the most humane option, it’s also best done by a veterinarian. ) 5. Rodent euthanasia using exposure to increasing concentrations of CO2 has come under scrutiny due more efficient method for euthanasia in small rodents in animal house facilities must still be identified [3,14]. CO 2 has many advantages as a euthanasia agent: it is a rapid depressant with well-established anesthetic effects, 3, 9 is readily available, is inexpensive compared with other euthanasia agents, poses Guidelines: Euthanasia of Mice and Rats. @article{Carbone2012AssessingCD, title={Assessing cervical dislocation as a humane euthanasia method in mice. 2 L/min was used to displace air at the rate of 21% per minute. In this experimental study, 10 female rats were randomly assigned to two equal groups and were There is currently insufficient evidence to advocate banning or replacing CO2 in the euthanasia of rodents; however, there are hints that alternative gases are more humane. 1258/0023677053739747 Corpus ID: 14005155; Carbon dioxide for euthanasia: concerns regarding pain and distress, with special reference to mice and rats @article{Conlee2005CarbonDF, title={Carbon dioxide for euthanasia: concerns regarding pain and distress, with special reference to mice and rats}, author={Kathleen M. 4 In the AVMA Guidelines, anesthetic overdose is recommended as an acceptable method of euthanasia. 0% CO 2 ). 8 min. CO 2 The aim of our work was to assess the impact of two methods of euthanasia (cervical dislocation versus inhalation of an overdose of isoflurane) on the quality of mouse oocytes. inhalation is considered an acceptable method of euthanasia for small rodents (e. Rats and mice Agents and Methods of Euthanasia by Species / Agents and Methods of Euthanasia by Species. The purpose of this study was to determine the time of CO2 exposure required to euthanize neonatal mice (0 to 10 days old). 3 Approved euthanasia methods will be specified in animal protocols. Selecting an appropriate, effective euthanasia agent is controversial. Title: Microsoft Word - I-IC-GU-214 Guidelines for CO2 Euthanasia in Rodents Euthanasia by isoflurane inhalation is an alternative method which allows the sacrifice of several mice at the same time with an Evaluating methods of mouse euthanasia on the oocyte quality: cervical dislocation versus isoflurane inhalation - Audrey Roustan, Jeanne Perrin, Anaïs Berthelot-Ricou, Erica Lopez, Alain Botta, Blandine Courbiere Chloroform inhalation is a common method of rodent euthanasia in Nigeria for research purposes. The AVMA Guidelines for the Euthanasia of Animals: 2020 Edition 1 describes that certain critical factors must be met to use CO 2 inhalation for euthanasia: “Carbon dioxide and CO 2 Although striving for a science-based report in which “only those methods for which reliable information could be obtained were included,”9 the Panel nonetheless approved, without citation of supporting data, “disarticulation of the skull and cervical vertebrae” as a method of mouse euthanasia. • This method of euthanasia may be used in small birds, poultry, mice, immature rats (weighing < 200 g) and rabbits. 17 Following this period, the mouse was removed from the induction chamber and cervical dislocation was performed as a secondary method of euthanasia. When using an anesthetic overdose to euthanize laboratory rodents, a secondary method of euthanasia is recommended to ensure that the apparent death is irreversible. Chloroform inhalation is a common method of rodent euthanasia in Nigeria for research purposes. Purpose. A combination of inhalation (primary) followed by a physical (secondary) method is recommended for all rodents as they, especially neonates, resist inhaled agents Background: There has been increased concern about the suitability of CO2 as a method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals Europe PMC is an archive of life sciences journal literature. 10 SUMMARY OF THE GUIDELINES LISTED IN THIS DOCUMENT Summary of Guidelines CCAC guidelines on: euthanasia of animals used in science 3. Author: Attending Veterinarian, Laboratory Animal Resources . 5%; Group B, inhaled sevoflurane at a concentration of 3%; and method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals may experience prior to loss of consciousness; time to loss Euthanasia method. We first explored the palatability of sugar-cookie dough with various flavorings added. 3 The optimal choice of euthanasia method for laboratory rodents depends on a number of factors, including the scientific goals of the study, the need to minimize animal pain and/or distress, applicable guidelines and laws, the training and proficiency of personnel, and the safety and emotional needs of the personnel performing the euthanasia. Euthanasia with carbon dioxide is not recommended for neonates, while cervical dislocation is an acceptable method [11], [12]. The Canadian Council for the Care of Laboratory Animals in its guidelines Euthanasia Method for Mice in Rapid Time-Course Pulmonary Pharmacokinetic Studies Download Article: Download (PDF 121. 5–18. Persons performing euthanasia should verify that the animals presented are those scheduled for euthanasia. Dr. A similar, yet, less pronounced trend was observed for JNK activity, whereas the choice of euthanasia method did not differentially impact p38 All currently accepted methods of euthanasia for laboratory mice involve some degree of stress, fear, anxiety, or pain. Since rodents can be particularly resistant to euthanasia by standard methods such as inhalation of CO 2 or gas anesthesia, IACUC policy requires that an approved secondary physical method of euthanasia (e. method of euthanasia. , voles, grass rats, jerboas, spiny mice) This lack of a euthanasia method that does not entail operational issues hampers the ability of veterinary professionals to provide appropriate guidance to scientists working with neonates. In a study on 400 articles that included surgical models in mouse and other species, the authors concluded that the animal A&A are We have assessed the time to death of foetuses after performing either indirect (dam euthanasia) or direct (via intraplacental injection--a new approach to euthanasia) euthanasia methods in order to determine a euthanasia method that is appropriate, ethical and efficient for the killing of mouse foetuses. Cervical dislocation: Euthanize rodents under 10 days old by one of the following procedures : 7. Stuckey JE 1 , Makhija SD 1 , Reimer DC 1 , Eswaraka JR. Light blue represents methods Barbiturate overdose. euthanasia. CO2, inhalant anesthetics) should be followed-up with a secondary technique (e. Mice also showed aversion to argon, CO, and the two General Concepts for All Methods • The method of euthanasia must be appropriate for the species and the age of the animal. , mice, rats, gerbils, hamsters, guinea pigs and degus) when properly administered. Evaluation of isoflurane overdose for euthanasia of neonatal mice. (3 authors) 1. Inhaled anesthetics - Nonflammable volatile anesthetic agents are effective for fetuses in utero. exsanguination or decapitation) for laboratory animals has been recommended to decrease pain or distress during the procedure. Set up a traditional spring trap or use a small-caliber, low-power gun. Skip to main content Skip to main navigation menu Skip to site footer Thus even at the highest flow rate tested mice experienced more than 30 s of dyspnea, suggesting other methods of euthanasia should be used when possible. 02-0. To define potential effects, we compared methods of lavage and euthanasia in uninjured mice and after a mild lung injury model (ozone). S6. 2, with time to cardiac arrest of 4:41 min in 1- to 7-day-old mice . In general, the IACUC will not approve CO2 as the sole method of euthanasia for mouse and rat fetuses and neonates. The UACC requires that a Primary (rendering death) and Secondary (confirmation of death) method of human killing be Euthanasia using gaseous agents in laboratory rodents A M Valentim1,2,*, S R Guedes3,*, A M Pereira3,* and L M Antunes1,2,3 Abstract Several questions have been raised in recent years about the euthanasia of laboratory rodents. 4 Subject: Euthanasia of Rodents via Physical Methods (Rat) Date Issued: August 21, 2012 Revision: 3. The euthanasia Mice and Rats . However, CO 2 is aversive for mice and likely causes Physical methods of euthanasia are also an appropriate means to assure death after euthanasia with anesthetics or CO 2 used as euthanasia agents. 19 We evaluated six methods of euthanasia, with and without anesthesia, in three age groups of mice: gravid mice (E14-20) and neonatal pups (P1-P7 and P8-P14). Cervical dislocation: Causes rapid death, but skill must be learned. 2,3,14,16 The AVMA Panel on Euthanasia provides guidance on the acceptable methods of euthanasia for rodents, including injectable and inhalant anesthetics. 5. Six different methods of anesthesia or euthanasia were evaluated for tissue collection (n = 8 mice per method of anesthesia). DOI: 10. 12 Euthanasia is considered to mean a ‘good death’ and therefore one without pain and suffering and, in some contexts, refers to a death that is in the animal’s interests (i. , voles, grass rats, jerboas, spiny mice) Exposure to carbon dioxide (CO2) is the most prevalent method used to euthanize rodents in biomedical research. 2% CO 2 ), bone-dry (> 99. 3 Although striving for a science-based report in which “only those methods for which reliable information could be obtained were included,”9 the Panel nonetheless approved, without citation of supporting data, “disarticulation of the skull and cervical vertebrae” as a method of mouse euthanasia. Isoflurane Inhalation 1. Current recommended flow rates range between 10 and 30% chamber vol/min and result in this basis, we argue that euthanasia methods that min-imize the duration of labored breathing should be con-sidered more humane. Aquatic invertebrates . This drug and route are recommended over inhalant anesthetics, carbon dioxide, and physical methods for ethical and aesthetic reasons as well as efficiency. , 2001; Klaunberg et al. To our knowledge, our study is the first to compare these two methods of mouse euthanasia on the oocyte quality. , requires that Institutional Animal Care and Use Committees (IACUCs) reviewing research animal activities determine that methods of euthanasia used will be consistent with the recommendations of the AVMA Guidelines for the Euthanasia of Animals unless a deviation is justified for scientific reasons in writing by the Background: There has been increased concern about the suitability of CO 2 as a method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals may experience prior to loss of consciousness; time to loss of consciousness; best methods for use of CO 2; and the availability of better alternatives. , 2004; Pritchett-Corning, 2009). This work is designed to evaluate the consequence of this method of sacrifice on the cerebellum and Neonatal mice have been reported to recover as long as after 30 min of exposure to CO 2 80 or isoflurane. • Exposure to 100% CO 2 for a minimum of 30 minutes. Chemical Methods. Policy: The ARC recommends anesthetic overdose or CO 2 asphyxiation for euthanasia of mice and The study demonstrated that exposure times of at least 3 min in carbon dioxide reliably result in irreversible euthanasia of mice but that exposure times of at least 10. Methods: Eighteen neonatal male mice were randomly divided into three groups. Physical methods of euthanasia that In the European Union (EU) millions of laboratory mice are used and killed for experimental and other scientific purposes each year. Jamie A Austin, 1 Y oungho Seo, 2 Anisha D Gandhi, 3 The PHS Policy, IV. Ernest Olfert, University of Saskatchewan University Animal Care Committee Standard Operating Procedure. PubMed. Confirmation of death is required immediately following a method of chemical (e. • For IACUC approval of this method in unanaesthetizedanimals, protocol participants must be properly trained and capable of consistently applying this technique humanely and effectively, as All currently accepted methods of euthanasia for laboratory mice involve some degree of stress, fear, anxiety, or pain. Finding a vein large enough to inject into is not an option, so the injection is given either directly into the heart or into the peritoneal cavity (belly). The present study aimed to assess the effects of two methods of laboratory animals’ euthanasia on PMCs in rats. Other methods outlined in the AVMA Guidelines on Euthanasia are acceptable with proper Euthanasia of Rodent Fetuses and Neonates - Acceptable Methods with Conditions. (hons. Similarly, ‘focused beam microwave irradiation’ method of euthanasia in mice significantly reduced brain mRNA expression levels, compared to euthanasia by CO 2 inhalation. 75 ml/10 g body weight (BW). MATERIALS . Euthanize many animals simultaneously is a prerequisite for conducting studies with large cohorts, especially for researches on female gametes. In most cases, overdose of injectable agents is the preferred method of euthanasia euthanasia. 86 Therefore, a secondary AVMA-accepted physical method of euthanasia (such as decapitation) should be performed to prevent revival. Note: All euthanasia practices will comply with current editions of the Guide for Care and Use of Laboratory Animals and the AVMA Guidelines for the The procedures listed below are suggested common methods for euthanasia of rodents. The present study demonstrated that intraplacental injection might be an effective euthanasia method that also allows for the preservation of anatomical and histological mouse foetal structures and avoids death by hypoxia. more efficient method for euthanasia in small rodents in animal house facilities must still be identified [3,14]. 5% of total body weight or 0. o With the animal(s) preferably still in its home cage, or in the euthanasia chamber that contains room air, gradually fill the home cage or the chamber with 100% CO 2 Intraperitoneal (IP) injection of sodium pentobarbital (PB) is an accepted method of euthanasia for mice. Euthanasia by isoflurane inhalation is an alternative method which allows the sacrifice of several mice at the same time with an anaesthesia, in the aim to decrease pain and animal distress. The ankle specimens were then immersed in 4% paraformaldehyde for fixation over Neonatal rats and mice (P1 to P14) have been shown to be more resistant to euthanasia by CO 2 overdose than adult animals, requiring prolonged exposure (Beaver et al. 1,2 Euthanasia of laboratory animals must be performed according to ethical standards, with trained personnel using appropriate techniques, equipment and reagents in order to induce a death as least painful and stressful as possible. When Naïve mice and mice with acute traumatic 'induced' brain injury showed varying levels of gene expression in hippocampus when evaluating the effect of euthanasia methods like isoflurane inhalation At the conclusion of experiments, the implementation of humane methods for rodent euthanasia is of paramount importance to uphold ethical standards for experiments on laboratory animals . Mice are commonly used in biomedical research, and euthanasia is an important part of mouse husbandry. Author information. Euthanasia using inhaled agents is considered to be a suitable aesthetic method for use with a large number of animals The use of sodium pentobarbital, injected intraperitoneally, for killing rodents is described as an acceptable technique by the AVMA and CCAC euthanasia guidelines. However, this method has important drawbacks, including the potential for pain or misinjection. This is because the procedure only requir es CCAC SECRETARIAT PROJECT TEAM Standards Team Ms. 1 (U. CO 2 is readily available, induces loss of consciousness in a rapid and reliable way with minimal personnel safety concerns and it has a relatively low cost [2,4,5]. B. Unless justified and approved in the Animal Use Protocol, a Primary method (rendering death) of humane killing must be followed by a Secondary (confirmation of The aim of our work was to assess the impact of two methods of euthanasia (cervical dislocation versus inhalation of an overdose of isoflurane) on the quality of mouse oocytes. Only trained personnel may perform euthanasia. For rodents, overdose of sodium pentobarbital is the most preferable euthanizing agent 4,5,6. 2. Background: There has been increased concern about the suitability of CO2 as a method for We found that relative to carbon dioxide asphyxiation, the other methods displayed significantly stronger ERK1/2 phosphorylation in select brain regions of male and female mice, with no pronounced sex difference. 1. First, the spine was palpated to assess whether palpable fractures of Here we compared 4 methods of spinal dislocation--3 targeting the cervical area (CD) and one the thoracic region--in regard to time to respiratory arrest in anesthetized mice. Decapitation: Useful when tissues must be free of euthanasia A number of different euthanasia protocols are used in animal research. to end pain and suffering). gas or injection) euthanasia ; Confirmatory methods for animals older than 14 days of age include, Confirmatory methods of euthanasia following an Introduction. Acceptable Methods a. Focused microwave irradiation. Note: All euthanasia practices will comply with current editions of the . 2 However, relatively little information is In 1972, the AVMA updated its recommendations on animal euthanasia. 3 Mouse, rat and hamster neonates up to 10 days of age method of euthanasia, but should not take precedence over animal welfare considerations. Euthanasia equipment i. 2 However, relatively little information is Corpus ID: 25294228; Assessing cervical dislocation as a humane euthanasia method in mice. Blogpost: UK animal research increases again. Liotti M, Brannan S, Egan G, et al. 19 7 methods of euthanasia commonly used in animals at UC Berkeley. The National Center for Scientific Research in France considers isoflurane inhalation to be the most appropriate method of euthanasia for rodents. Procedures: **Unless described in the animal use protocol and approved by the UACC, all physical methods of humane killing must be performed under anaesthesia following the appropriate anaesthesia SOP (7. The euthanasia method used must be described in the approved ACAP. The Canadian Council for the Care of Laboratory Animals in its guidelines Background: There has been increased concern about the suitability of CO2 as a method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals There are specific training methods for rodents that align with the principles of refinement, focusing on gentle handling, positive reinforcement, and reduced stress during conducting a ‘preliminary study’ is a good way to determine the most appropriate euthanasia method for the specific study and groups of rodents committee is provided with adequate justification for the use of the method and references that support its use in the proposed circumstances. The AVMA Guidelines on Euthanasia require anesthesia for It can be difficult, however, to apply mammalian methods of euthanasia to species with unique physiology such as reptiles. CD was first approved for mouse euthanasia in 1972 b Cervical dislocation is a commonly used method of mouse euthanasia. Recommended Standard Methods of Blood Collection: Mice Method Maximum volume (per sample) Anesthesia Required Description Special Considerations • Ensure euthanasia of the animal post-procedure. As PB rapidly causes sedation and loss of consciousness, Physical methods of euthanasia are also an appropriate means to assure death after euthanasia with anesthetics or CO 2 used as euthanasia agents. 5 SC, IM The optimal choice of euthanasia method for laboratory rodents depends on a number of factors, including the scientific goals of the study, the need to minimize animal pain and/or distress, applicable guidelines and laws, the training and proficiency of personnel, and the safety and emotional needs of the personnel performing the euthanasia. Guide for Care and Use of Laboratory Animals . 011115 The mouse is the most commonly used model in As euthanasia was the intended endpoint, the mouse was then immediately transferred to a chamber pre-filled with 80% CO 2 for a minimum of 5 min, following UBC guidelines. Ronald Charbonneau, Centre Hospitalier de l'Université Laval Dr. Cardiovascular, behavioral, and activity data did not consistently or significantly support isoflurane or pentobarbital-phenytoin euthanasia as less stressful methods than CO2, and it is concluded that using CO2 with or without isofLurane is an acceptable euthanasia method. , mice, rats, Mouse, Rat, and Hamster Neonates up to 10 days of age: Acceptable methods for euthanasia include: injection of chemical anesthetics (e. Examples of methods for rodents that are acceptable and acceptable with conditions are given in Table 36. For studies in which death of the animal can be anticipated or is an inevitable part of the protocol, the investigator should specify the end point of the experiment and alternative situations in which termination of the experiment would be mandatory to avoid distress. Recommended euthanasia techniques for small mammals (e. Thus, the use of CO 2 as a method of euthanasia could result in an experimental artifact that could compromise results when studying labile biologic processes. Section 1: Definitions - Details for conduct of procedures can be found in Supplemental SOP information. Non-physical methods of euthanasia utilizing inhalant agents (e. Light blue represents methods permitted or recommended for use, dark blue represents methods where additional permissions are required, navy blue represents methods not permitted or recommended. In general, the experimental animals are euthanatized by physical or inhalation methods. Euthanasia should be conducted professionally and compassionately by skilled persons using means that are optimal for the circumstances. Euthanasia Method for Mice in Rapid Time- Course Pulmonary The University of Washington IACUC notes that larger mammals appear to be distressed by CO 2 euthanasia. Euthanasia using inhaled agents is considered to be a suitable aesthetic method for use with In the AVMA Guidelines, anesthetic overdose is recommended as an acceptable method of euthanasia. In all cases, tissue collection procedures were initiated after animals had either been euthanized or were under deep anesthesia and unresponsive to all stimuli. Cervical dislocation for rats weighing less than 200 grams and all mice after sedation (unless otherwise scientifically justified to U. 9% CO 2 ), or industrial (> 99. Substantial resources have been allocated to the evaluation of this question in postweanling rodents, 9, 11, 12 resulting in revisions to the AVMA Guidelines on Euthanasia in 2020. A major benefit of This chapter is designed to give a general overview of euthanasia and necropsy techniques that may be used for laboratory rabbits, guinea pigs, rats, mice, and other rodents. ), the university of british columbia, 2011 a thesis submitted in partial fulfillment of the requirements for the degree of master of science in the faculty of graduate and postdoctoral studies Euthanasia Method for Mice in Rapid T ime- Course Pulmonary Pharmacokinetic Studies Adam R Schoell, 1,* Bruce R Heyde, 2 Dana E Weir , 1 Po-Chang Chiang, 3 Yiding Hu, 2 and David K T ung 3 Table 1. Methods of Confirmation of Euthanasia. Table 1. Based on 'Euthanasia of Animals Used for Scientific Purposes' ANZCCART 2001. The objective of this prospective, randomized, blinded study was to determine whether intrahepatic CO 2 administration euthanasia method had an in-between foetal time value to death. Naïve mice and mice with acute traumatic ‘induced’ brain injury showed varying levels of gene expression in hippocampus when evaluating the effect of euthanasia Background: There has been increased concern about the suitability of CO 2 as a method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals may experience prior to loss of consciousness; time to loss of consciousness; best methods for use of CO 2; and the availability of better alternatives. Contents The SWMS Methods of Humane Euthanasia in Micecontains the following sections: o Legislation University Policy Local Policy Safe Work Method Statement Personal Protective Equipment Required Hazards and Controls Before Work Commences The use of anesthetics in association with a confirmatory method of euthanasia (e. 19 Since then, with no studies of CD in General recommendations for the process of euthanizing rodents used in research include the maintenance of stable groups, reduction of transport prior to the euthanasia process, and selection of euthanasia methods that minimize pain and distress. 2%, but concentrations of greater than 30% are needed to render them unconscious. R. Google Scholar. Mice were euthanized individually in a standard mouse IVC (no. Euthanasia procedures should be swift and minimize pain and distress, aligning with the principles of 3R-Refinement. appears to be an alternative method of euthanasia com-pared with cervical dislocation. , rodents), birds, and ectotherms as well dislocation is a humane technique for euthanasia of mice, other small rodents, and rats weighing less than 200 grams. Carbon dioxide generated from dry ice is An effective and pain-free killing method is required to achieve the goal of euthanasia, a “good death”. The euthanasia method selected for use must be described in detail in your approved Animal Study Proposal. Mouse and rat fetuses and neonates (up to 14 days of age) show extraordinary tolerance for CO2 exposure, with up to 50 Our results indicate that the most common method of euthanasia for laboratory mice – exposure to CO 2 – is at least moderately aversive to mice. 7,16 An additional recommendation is that the wellbeing of rodents is improved when the rodent is euthanized SWMS 1. Date: February 28, 2022. Methods of euthanasia: (select all that apply) Primary methods: Agent/Method Name Dosage (in mg/kg if possible) Route Carbon Dioxide (CO2) 30-70% chamber volume per minute (Flow meter: Rat: 6-8 L/min; Mouse cage: 4-6 L/min) Inhalation Isoflurane Open drop method or Precision Vaporizer 5% Inhalation . This review examines previous studies on CO2 euthanasia and presents the current body of knowledge on the subject. 3. Euthanasia Guidelines In their 2020 report, the AVMA Guidelines for the Euthanasia of Animals: 2020 Edition reviewed the current literature and concluded that “the practice of immersion, Guidelines for Euthanasia of Rodents Using Carbon Dioxide. Minimization of potential pain and distress of rodents undergoing euthanasia is a touchstone of veterinary clinical medicine. A major benefit of Although controversially discussed, the use of carbon dioxide (CO2) is still permitted for killing rodents according to the Directive 2010/63/EU. Department of Agriculture 2002), define euthanasia as the This document, the CCAC guidelines on: euthanasia of animals used in science, has been developed by the ad hoc subcommittee on euthanasia of the Canadian Council on Animal Care (CCAC) Guidelines Committee. Seymour TL, Nagamine CM. In Practice: Advanced in vivo Modeling in the Mouse The word euthanasia comes from the Greek terms eu (good) and thanatos (death). 3 c: Immersion in anesthetic solution (magnesium salts, clove oil, eugenol, ethanol) S6. This cage was not bedded and was not a home cage. Refer to NHGRI Guideline 01. Location: Queen’s University Responsibility: Principal Investigators (PI), Research Staff, Veterinary Staff Purpose: The purpose of this Standard The method of euthanasia should be appropriate for the species and the age of the animals. Animal welfare, ethics, euthanasia methods, mice, rats, 3Rs Date received: 4 February 2022; accepted: 6 April 2022 Why consider killing methods for use in laboratory rodents? Rodents remain the most widely used species for scien-tific research due to their small size, low cost, rapid sexual maturity and scope for genetic manipulation. , rabbits). The CO2 method causes death to An experienced veterinarian, blinded to operator and euthanasia method, examined each mouse within 2 h of euthanasia. . CO2 Compressed CO2 gas is the only recommended source of CO2 for euthanasia. CO 2 Below are descriptions of standard acceptable euthanasia methods: RODENTS. Others choose CD to assure death in mice treated with injected or inhaled euthanasia agents. Research investigators often choose to euthanize mice by cervical dislocation (CD) when other methods would interfere with the aims of a research Euthanasia Method in Mice. and 3:52 min in 8- to 14-day-old mice. This study aimed to evaluate pain associated with six methods of euthanasia in Wistar rats (injectable, inhalational, and physical), by applying the Rat Grimace Scale (RGS), comparing the scores, This is Appendix 2 of the Code of Practice for the Housing and Care of Laboratory Mice, Rats, Guinea Pigs and Rabbits. According to The method of euthanasia should be appropriate for the species and the age of the animals. Neonates over 14 days of age may be euthanized as per the guidelines for adults. For mice, some researchers advocate physical methods such as cervical dislocation — a skilled breaking of the neck — but this is difficult to achieve with conscious rats because of their size. General recommendations for the process of euthanizing rodents used in research include the maintenance of stable groups, reduction of transport prior to the euthanasia process, and selection of euthanasia methods that minimize pain and distress. NOTES: Total blood volume of a mouse is 75 ml/kg or 7. Group A, inhaled sevoflurane at a concentration of 1. Several questions have been raised in recent years about the euthanasia of laboratory rodents. 19 7 In 1972, the AVMA updated its recommendations on animal euthanasia. Of the 81 mice Methods: The International Association of Colleges of Laboratory Animal Medicine (IACLAM) convened a taskforce to examine the evidence for adverse welfare indicators in Following euthanasia, the ankles were excised, and soft tissues were removed from the mice. , pentobarbital), decapitation or cervical Overview of killing methods from published euthanasia guidelines for adult laboratory rodents across scientifically advanced countries. For the SF method, mice were placed in the cage, and CO 2 at a flow rate of 1. For mice, acceptable confirmatory methods after CO 2 exposure include:. We used adult Swiss Webster (male) mice to evaluate injectable and We have assessed the time to death of foetuses after performing either indirect (dam euthanasia) or direct (via intraplacental injection – a new approach to euthanasia) euthanasia methods in order to determine a euthanasia method that is appropriate, ethical and efficient for the killing of mouse foetuses. decapitation, cervical dislocation, thoracotomy, immediate harvest of critical The American Veterinary Medical Association (AVMA) Panel on Euthanasia defines euthanasia as “a good death that occurs with minimal pain and distress,” 1 and there are many methods acceptable In 1972, the AVMA updated its recommendations on animal euthanasia. The preferred In particular, investigators use various methods for BAL and euthanasia; however, the impact of these methods on assessments of injury and inflammation is unknown. 31 That study 31 found that the intraperitoneal injection of 0. For adult animals, euthanasia with barbiturates and carbon dioxide are recommended methods [11], [12]. AVMA Euthanasia Guidelines classifications: Euthanasia methods are classified in the Guidelines as acceptable, acceptable with conditions, and unacceptable. The normal method used for dogs and cats is by giving an injection of a euthanasia drug directly into a vein, but this cannot be done on a rat or mouse due to its size. Currently, pulmonary pharmacokinetics studies use a variety of euthanasia techniques, including CO 2 exposure, intraperitioneal barbiturate overdose, cervical dislocation, and decapitation. The title of the document has been changed from “Mouse and Rat Euthanasia Procedures” to “Rodent Euthanasia Procedures” to reflect the inclusion of other rodent species (e. 1, Table 36. Pentobarbital and Phenytoin (Euthasol) Chemical euthanasia with an overdose of intravenous pentobarbital is the preferred euthanasia method for mammals with practical vascular access (e. 2012. Larry Carbone, 1,* Elizabeth T Carbone, 1 Elizabeth M Yi, 1 Diana B Bauer, 1 Krista A Lindstrom, 1 John M Parker, 1. Introduction. Brain responses associated with consciousness of breathlessness (air hunger). This secondary method usually is accomplished through the collection of tissues that are required to complete the research project. g. Rodent euthanasia using exposure to increasing concentrations of CO2 has come under scrutiny due Introduction. gas or injection) euthanasia ; Confirmatory methods for animals older than 14 days of age include, Confirmatory methods of euthanasia following an Figure 2: Experimental design: Our scheme was based on the assessment of the presence of pain and suffering during euthanasia induction, the time of death confirmation and the interference in the biological systems of each methodology used through the collection of blood and organs. The second study examined the same three measures of insensibility during the isoflurane method of euthanasia, with the aim of identifying when it is safe to switch to a high flow rate of CO 2, In this study, we examined 3 commonly used euthanasia methods in mice: intraperitoneal injection of pentobarbital-phenytoin solution, CO2 inhalation, and isoflurane anesthesia followed by CO2 There is insufficient evidence to permit an unbiased assessment of the effect of CO2 inhalation during euthanasia on welfare indicators in laboratory mice and rats, and it is likely that repeated exposures to carbon dioxide inhalation are aversive to adult rats and mice. In this study, mice were euthanized with one of 3 grades of CO 2: USP medical (> 99. 1258/la. 7, 16 An additional recommendation is that the wellbeing of rodents is improved when the rodent is euthanized method of euthanasia of neonatal mice was inhalation of CO. 1 "Euthanasia of Rodents" and the AVMA Panel on Euthanasia for additional information. I‐IC‐GU‐202 Euthanasia of Mouse and Rat Fetuses and Neonates. Times to recumbency and cessation We evaluated six methods of euthanasia, with and without anesthesia, in three age groups of mice: gravid mice (E14-20) and neonatal pups (P1-P7 and P8-P14). Although controversially discussed, the use of carbon dioxide (CO2) is still permitted for Research investigators often choose to euthanize mice by cervical dislocation (CD) when other methods would interfere with the aims of a research project. Ho The term ‘killing’ refers to any intentional act or process that results in the death of an animal. ” 31 Control groups were not included in the study, and specific This article explores the ethics of animal euthanasia in wildlife rehabilitation and the methods commonly used to perform the task. CO 2 asphyxiation under isoflurane anesthesia followed by decapitation. To date, however, there are few empirically based studies that have investigated the extent to which different methods may influence the reliability and generalizability of results across studies. Euthanasia chamber with CO2 gas supply ii. kuspgbf qqlk hksrw tbs yahhkoh bjdfl vmzdtucc bnghtl xbljh yyrrwdp