Safe Work Method Statement. Mouse Anaesthesia and Analgesia

Transcription

1 Flinders University Safe Wrk Methd Statement Muse Anaesthesia and Analgesia Cllege f Medicine and Public Health Animal Facility SWMS Number RA Number RA Scre SWMS- 1.3 RA- 1.3 MEDIUM Cntact Persn SWMS prepared by AWC Apprval Date Review Date Rxanne Cllingwd Erin Plews and Lewis Vaughan 15 August 2017 August 2022 Cntents The SWMS Muse Anaesthesia and Analgesia cntains the fllwing sectins: Legislatin University Plicy Lcal Plicy Safe Wrk Methd Statement Persnal Prtective Equipment Required Hazards and Cntrls Befre Wrk Cmmences General Infrmatin NHMRC/ Flecknell P 2015/ AWO Anaesthesia and Analgesia Guidelines Anaesthesia Settings and Setup Inductin and Maintenance f Anaesthesia Mnitring Depth f Anaesthesia Recvery frm Anaesthesia and Sedatin Medicatins Assciated with Anaesthesia and Surgery SWMS 1.3 Muse Anaesthesia and Analgesia Page 1 19/12/2016

2 Legislatin Australian Cde fr the Care and Use f Animals fr Scientific Purpses 8 th Ed. Animal Welfare Act 1985 Animal Welfare Regulatins 2012 Gene Technlgy Act 2000 (the Act) Gene Technlgy Regulatins 2001 Wrk Health and Safety Regulatins 2012 University Plicy Wrk Health and Safety Plicy 2013 Respnsible Cnduct f Research Plicy 2016 NHMRC Guidelines Lcal Plicy Use f the Cllege f Medicine and Public Health Animal Facilities by all staff and researchers f the Cllege f Medicine and Public Health, Flinders University, is subject t awareness f, and adherence t the fllwing: Research Invlving Animals: The University hlds a licence fr the use f animals fr teaching and research purpses. T satisfy the requirements f the licence, anyne wishing t undertake teaching and research using animals must submit a prpsal t the Animal Welfare Cmmittee (via the Animal Ethics Review Sub- Cmmittee. N wrk with animals may cmmence until written apprval has been received frm the Animal Welfare Cmmittee. Standardised applicatin frms fr Research and Teaching can be fund n the Flinders University website listed belw. It is yur respnsibility t regularly check this site fr updates t guidelines, frms etc All staff and students invlved in animal research must cmplete Animal Ethics Online Training (AEOT) and must als regularly attend Animal Researcher Infrmatin Sessins (ARIS). All persnnel wrking with Genetically Mdified Animals (GMO) r wrking with in a PC1 r PC2 facility must attended a Bisafety Training Day every 3 years. Safe Wrk Methd Statement Refer t Risk assessments, Safe Wrk Methd Statement fr chemicals, prcesses and plant equipment where apprpriate. All prjects must have an accmpanying Risk Assessment signed by the Animal Facility Manager Related SWMS dcuments include: SWMS 1.0 Muse- Sexing, Handling, Restraint and Ear Ntching RA 1.0 Muse- Sexing, Handling, Restraint and Ear Ntching SWMS 1.7 Muse -Transprtatin RA 1.7 Muse Transprtatin SWMS 1.9 Muse PC1, PC2 and Infectius Cntainment Husbandry RA1.9 Muse PC1, PC2 and Infectius Cntainment Husbandry SWMS Emergency Cntingency RA Emergency Cntingency Glves Gwn Mask Hair Net Persnal Prtective Equipment Required SWMS 1.3 Muse Anaesthesia and Analgesia Page 2 19/12/2016

3 She Cvers Hazards and Cntrls Anaesthetic Inhalatin may prduce health damage. Cumulative effects may result fllwing expsure. D nt breathe gas/fumes/vapur/spray. Use nly in well ventilated areas and use scavenge systems. Remve all cntaminated clthing immediately if spilt n bdy. If a spill ccurs, vacate the area, clse drs, ring 33#, alert thers in the vicinity. Animal bites and scratches are prevented by training, demnstratin f cmpetency, adhere t SWMS. Needle Stick- DO NOT recap needles, dispse immediately int sharps cntainers, adhere t the relevant SWMS. Befre Wrk Cmmences Ensure that yu are aware f the lcatins f the fllwing: Spill Kit Fire Extinguisher Eye Wash Exits Risk Assessment and SDS (Safety Data Sheet) - Ensure that yu have read and understd fr all the substances being used. Equipment Check fr safety and electrical cmpliance Ensure that yu have read and understd the Risk Assessment and Safe Wrk Methd Statements Obtain training befre using any equipment List Of Equipment Needed Isflurane Inductin Chamber Filler Key Heat Mat Anaesthetic machine Heat mat SWMS 1.3 Muse Anaesthesia and Analgesia Page 3 19/12/2016

4 General Infrmatin All prcedures are t be perfrmed by trained cmpetent staff. Training is available frm senir animal huse staff r Animal Welfare Officer. Evidence f training is available in the Staff Training Needs Analysis and the Cmpetency Skills Register administered by the Research Services Office. Wmen f childbearing age shuld be warned that expsure t Isflurane may have an effect n fertility and fetal develpment. Wmen wh are planning pregnancy, r wh are pregnant, may wish t request t be remved frm tasks invlving direct expsure, such as filling Isflurane machine. SWMS 1.3 Muse Anaesthesia and Analgesia Page 4 19/12/2016

5 NHMRC/ Flecknell P 2015/ AWO Anaesthesia and Analgesia Guidelines Anaesthesia Drug Dse Rute Isflurane 3-5% Inductin Inhalatin 1-4% Maintenance Inhalatin Servflurane 5-8% Inductin Inhalatin 2.5-4% Maintenance Inhalatin Ketamine mg/kg IP Aceprmazine, r mg/kg IP Buprenrphine r 0.05 mg/kg IP Butrphanl mg/kg IP Ketamine mg/kg IP Medetmidine mg/kg IP Reverse medetmidine with atipamezle 1 mg/kg IP r SC Ketamine + 100mg/kg IP Xylazine 10 mg/kg IP Reverse xylazine with atipamezle 1 mg/kg SC Ketamine + 100mg/kg IP diazepam 5 mg/kg IP Pentbarbitne mg/kg (adult) IP 10-20mg/kg (yung) IP Alphaxalne mg/kg IV Analgesia Drug- NSAID Dse and frequency Rute Carprfen 5 mg/kg, 24 hurly SC Melxicam 1-5 mg/kg, 24 hurly SC r per s Drug- Opiid Dse and frequency Rute Buprenrphine mg/kg, 12 hurly SC Buprenrphine ral 1 mg/kg in Nutella (give 125 mg t a 25 g muse f 0.2 mg/g nut paste) 24 hurly as per Abelsn et al 2012 PO Butrphanl mg/kg, 4 hurly SC Reverse µ piates with butrphanl mg/kg SC Reverse Opiate and Opiid with Nalxne 3mg/kg IP, IM r IV Sedatin Acetylprmazine nly, r with mg/kg IP Butrphanl r mg/kg IP Buprenrphine 0.05 mg/kg IP Lcal Anaesthesia (Animal Welfare Officer Recmmendatins) Ligncaine Dilute t 0.5%, recmmended incisinal vlume is 50 µl, Waite et al, BJA, 104 (6): , maximum dse is 10 mg/kg, fast Infiltratin nset shrt acting (<1 hur) Bupivacaine Dilute t 0.25%, recmmended incisinal vlume is 50 µl, Waite et al, BJA, 104 (6): , maximum dse is 3 mg/kg, ttal dse q1-6h infiltratin and epidural and q8-24h plexus r nerve Infiltratin Mix 1% ligncaine with 0.25% bupivacaine as a 50:50 mixture. Dilute Ligncaine/bupivacaine mix this mixture with an equal quantity f water fr injectin. Infiltrate the incisin with a maximum f 50µL (Flecknell, 2015) Infiltratin SWMS 1.3 Muse Anaesthesia and Analgesia Page 5 19/12/2016

6 Suggested indicatins and prtcls fr researcher/technician training by the AWO and Animal Facility staff: - Mice sedatin fr mderately invasive prcedures such as gavage, cheek bleed training etc, either: r: acetylprmazine/butrphanl at 2.5 t 5 mg/kg -1 /1.0 t 2 mg/kg -1 i.p r s.c Fr the maximal dse mix 1.25 mls f acetylprmazine ( 2mg/mL) with mls f butrphanl (10 mg/ml) and add sterile Nrmal Saline fr injectin up t 5 mls dse at 0.1 ml/ 10 g bdyweight. acetylprmazine/buprenrphine at 2.5 t 5 mg/kg -1 /0.05 mg/kg -1 i.p r s.c Fr the maximal dse mix 1.25 mls f acetylprmazine ( 2mg/mL) with 0.08 mls f Temgesic (300 mcg/ml) and add sterile Nrmal Saline fr injectin up t 5 mls dse at 0.1 ml/ 10 g bdyweight. - Mice sedatin alternative t the abve using xylazine at 10 mg/kg-1 Mix 0.5 ml f xylazine (20 mg/ml) and add sterile Nrmal Saline fr injectin up t 10 mls dse at 0.1 ml / 10 g bdyweight. - Muse nn-recvery anaesthesia, fr invasive prcedures, after previus acetylprmazine/butrphanl r acetlyprmazine/buprenrphine sedatin, Zletil at 40 t 60 mg/kg-1 by i.p r s.c injectin Mix 0.4 t 0.6 mls f Zletil ( 100 mg/ml) and add sterile Nrmal Saline fr injectin up t 10 mls dse at 0.1 ml / 10 g bdyweight. - Muse nn-recvery anaesthesia fr invasive prcedures withut pre-medicatin, Zletil/medetmidine at 40 t 60 mg.kg -1 Zletil /0.5 mg.kg -1 medetmidine i.p r s.c injectin Mix 0.4 t 0.6 mls f Zletil (100 mg/ml) and 0.5 mls f Dmitr ( 1mg/mL) and add sterile Nrmal Saline fr injectin up t 10 mls dse at 0.1 ml / 10 g bdyweight. - Muse recvery anaesthesia f shrt term duratin, ketamine/xylazine at 100 mg.kg -1 / 10 mg.kg -1 xylazine by i.p r s.c injectin Mix 1 ml f ketamine (100 mg/ml) and 0.5 mls f xylazine (20 mg/ml) and add sterile Nrmal Saline fr injectin up t 10 mls dse at 0.1 ml / 10 g bdyweight. NOTE: A Permit t Pssess Pisns and Cntrlled Drugs must be btained frm SA Health Drugs f Dependence Unit befre being able t use Schedule 8 Cntrlled Drugs. All drugs must be kept in a lckable cabinet when nt in use, and a register f use with balance remaining maintained. Anaesthesia Settings and Setup Check the level f Isflurane and tp up using the methd belw if required. Methd: 1. T fill the vapriser yu must have an Isflurane filler key, as this is the nly way the vapriser can be filled. SWMS 1.3 Muse Anaesthesia and Analgesia Page 6 19/12/2016

7 2. Attach the key filler t the tp f the Isflurane bttle by matching up gaps n the key with the cllar n the bttle, and screw n firmly. 3. Open the fill prt by unding the screw and remving the prt plug, and place the key in the prt. Once fully inserted, lck the key in place by tightening the screw. 4. Raise the Isflurane bttle abve the level f the prt t fill the vapriser. The Fill windw shws the level f Isflurane in the vapriser. T cease filling the vapriser, lwer the bttle f Isflurane belw the level f the prt when the level f Isflurane is level with the the tp line in the fill windw. 5. Release the key lsening the screw, remve the filling key, replace the fill prt plug, and retighten the screw. Remve key filler frm the Isflurane bttle, and recap the bttle firmly. Gaseus Anaesthesia: Isflurane Inductin setting range is between 2-5%. A rutine inductin setting is 3% fr mst animals. Maintenance setting range is 1-4%. Mst animals can be maintained between 1.5 and 3%. Oxygen flw rate (flw meter): Fr inductin in chambers - the xygen flw rate is recmmended t be set t fill the chamber as rapidly as pssible. When using nn-rebreathing mask systems the flw rate is recmmended t be a minimum f 500 mls/minute. Inductin and Maintenance f Anaesthesia NOTE: An anaesthetised animal must never be placed int a cage with a nnanaesthetised animal. NOTE: All animals must be cntinually mnitred until fully recvered and mbile fllwing prcedures. The Midget 3 anaesthetic machine can be used as an pen circuit, nn-re-breathing system with a mask fr maintenance anaesthesia f small labratry animals. 1. Rdents shuld be placed n warm mats fr any anaesthetic lasting mre than 5 minutes. Turn n the warm mat quarter f an hur befre it is required. Ensure it is insulated frm the table tp. SWMS 1.3 Muse Anaesthesia and Analgesia Page 7 19/12/2016

8 2. Turn n the xygen at the wall, if using the mains xygen plumbed int the theatres, r if using an xygen bttle, check that enugh pressure remains in the bttle. Set the flw meter t deliver the maximum flw rate pssible t the chamber. Ensure the scavenger is attached and turn n at the crrect setting. 3. Ensure flw is directed t the chamber r mask. 4. Place the rdent in the chamber and ensure it is sealed. T prvide pre-xygenatin, allw the animal t stay in the chamber in the xygen flw fr up t 5 minutes prir t turning n the vapriser. Mice receiving injectable anaesthetics shuld be expsed t 100% xygen in the latter manner immediately after injectin until prcedures cmmence. 5. T induce anaesthesia, turn n the Isflurane vapriser t between 2.5 t 5%. 6. Once the animal is deemed t be sufficiently anaesthetised, transfer the animal t a nnrebreathing mask-circuit and set the isflurane t between 1.5 t 2.5 %. An phthalmic intment, such as Lacrilube, shuld be applied t the eyes f the animal and 1 t 1.5 mls f Sterile Nrmal Saline fr Injectin administered by the subcutaneus r peritneal rute unless these agents are cntraindicated with regard t the research mdel r clinical cnditin f the animal. 7. Cntinue t mnitr the animal and adjust the vapriser as necessary. 8. On cmpletin f the prcedure, turn the Isflurane ff and keep the animal n 100% xygen until vluntary mvement makes maintaining supplemental xygen impssible withut animal restraint. If an animal has received an injectable anaesthetic, prvide it with 5 minutes f 100% xygen after the end f the prcedure. Prviding pre and pst xygenatin will reduce the incidence f adverse events during anaesthesia, and is recmmended as rutine practice. NOTE: The suctin shuld be cnnected t the mask t minimise leakage f gases and t reduce carbn dixide build up. When using a mask, the suctin is t be kept lw s as nt t remve the gas befre the animal can inhale it. Anaesthetic Machine Nse Cne Isflurane Dial Adjust accrding t depth f anaesthesia required Depress t flush system with 100% xygen Oxygen flw Meter Gas utlet- cnnect t nse cne r inductin chamber Gas Inlet Exhaust Gas Cnnect t scavenge system SWMS 1.3 Muse Anaesthesia and Analgesia Page 8 19/12/2016

9 Mnitring Depth f Anaesthesia NOTE: Muse shuld be kept n a heat mat during the prcedure and kept in an ambient temperature f C until the animal recvers righting reflex. 1. Inductin - Animal will begin t lse its righting reflex. 2. Adequate Anaesthesia - Tail Pinch and pedal withdraw reflex (lack f mvement fllwing firm pinch f the tail and the webbing between the feet digits). A Firm pinch with plain frceps, which des nt break the skin r cause any deep tissue damage, is sufficient t shw if the animals is t light. Any bserved mvement (withdrawing the tail r ft) indicates that the animal is nt sufficiently anesthetized t d surgery. Respiratin rate shuld be regular Mucus membranes and expsed tissues shuld be bright pink t red. 3. Anaesthesia Inadequate - Any mvement f the muse whilst under anaesthesia indicate inadequate depth, s therefre slightly increase the amunt f Isflurane delivered. Rapid, shallw respiratins usually indicate the animal is t "light". 4. Anaesthesia t deep - Respiratin Rate may be deep, shallw, decreased r irregular. Mucus membranes and expsed tissues shuld be bright pink t red if the clur changes t grey r blue clur anaesthesia is t deep. Spasmdic gasps with dilated pupils and/r blue/grey muzzle and extremities is assciated with anxia and excessively deep anaesthesia. Decease r turn ff the Isflurane being delivered and flush the circuit with pure xygen. Recvery frm Anaesthesia and Sedatin Fr recvery purpses, if animals have received injectable anaesthesia/sedatin, r have nt regained full mbility within 15 minutes after the terminatin f gaseus anaesthesia, the fllwing actins shuld be taken: 1. Place tw t three fd cubes int a cup cake cntainer, and add sufficient water t sak the fd, and leave this n the cage substrate at the ppsite end f the cage t the nest. 2. Administer a vlume f bdy-temperature Nrmal Saline fr Injectin, equivalent t 5% f bdyweight, by subcutaneus injectin with a maximum vlume f 0.5 mls in each injectin site. 3. Place the cage n a thermstatically cntrlled warm mat with the mat in cntact with half f the external flr f the cage. Leave the cage n this warm mat until the animals are capable f independent purpseful mvement. 4. Mnitr clinical signs f recvery and cmplete Clinical Recrd Sheets as specified in the AWC apprved prject details. 5. If animals have nt recvered full mbility by clse f business, cntact the Animal Welfare Officer and fllw veterinary advice. SWMS 1.3 Muse Anaesthesia and Analgesia Page 9 19/12/2016

10 Medicatins Assciated with Anaesthesia and Surgery Surgical practice shuld be cnducted, as far as pssible, in an aseptic manner. Antibitics shuld nt be used as an alternative t aseptic technique. The use f prphylactic antibitics shuld be restricted t single injectins f shrt-acting narrw t bradspectrum agents, r lng-acting narrw spectrum agents. Antibitics which are recmmended t prevent r treat bacterial infectins assciated with surgery are: (i) sluble ampicillin at 50mg/kg by subcutaneus r intraperitneal injectin; (ii) enrflxacin (25 mg/ml r less) at 10 mg/kg. Where pssible, based n AWO advice, alternatives t enrflxacin shuld be used; r (iii) alternative antibitics as recmmended by the AWO. These agents shuld be administered prir t the start f surgery, in rder t btain therapeutic tissue levels f the drugs in the peri and pst-perative perids. If pstsurgical infectin ccurs, the AWO shuld be cnsulted fr treatment advice. SWMS Review This SWMS currently applies t the animals hused in the Cllege f Medicine and Public Health Animal Facility. This SWMS will be reviewed 5 yearly, but als updated mre frequently as plicies, techniques and animal care requirements change. Psitin Name Cntact Details Manager Animal Facility Rxanne Cllingwd rxanne.cllingwd@flinders.edu.au Animal Welfare Officer Lewis Vaughan aw@flinders.edu.au Useful References - Guidelines t prmte the wellbeing f animals used fr scientific purpses, NHMRC, Flecknell PA (2015). Labratry Animal Anaesthesia, Elsevier KSP Abelsn, KR Jacbsen, R Sundbm, O Kallikski and Hau J, (2012), Vluntary ingestin f nut paste fr administratin f buprenrphine in rats and mice, Labratry Animals, 46: Quesenberry K and Carpenter J, Ferrets, Rabbits and Rdents (2012) Elsevier, Missuri, USA, 3 rd ed, chapter 31. Waite A et al (2010), Clinically relevant dses f lidncaine and bupivacaine d nt impair cutaneus wund healing in mice, British Jurnal f Anaesthesia, 104 (6): SWMS 1.3 Muse Anaesthesia and Analgesia Page 10 19/12/2016

11 Any questins regarding the abve guidelines and any technical advice/ assistance required can be directed t Animal Facility Manager r the Animal Welfare Officer. Versin 8 SWMS 1.3 Muse Anaesthesia and Analgesia Page 11 19/12/2016