Safe Work Method Statement. Rat Anaesthesia and Analgesia

Transcription

1 Flinders University Safe Wrk Methd Statement Rat Anaesthesia and Analgesia Cllege f Medicine and Public Health Animal Facility SWMS Number RA Number RA Scre SWMS- 2.3 RA- 2.3 MEDIUM Cntact Persn SWMS prepared by AWC Apprval Date Review Date Rxanne Cllingwd Rxanne Cllingwd and Lewis Vaughan 15 August 2017 August 2022 Cntents The SWMS Rat 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 Anaesthesia and Analgesia Guidelines Anaesthesia Settings and Setup Inductin and Maintenance f Anaesthesia Intubatin Mnitring Depth f Anaesthesia Medicatins Assciated with Anaesthesia and Surgery Legislatin Australian Cde fr the Care and Use f Animals fr Scientific Purpses 8th Ed. Animal Welfare Act 1985 Animal Welfare Regulatins 2012 Gene Technlgy Act 2000 (the Act) Gene Technlgy Regulatins 2001 SWMS 2.3 Rat Anaesthesia and Analgesia Page 1 19/12/2016

2 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 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 SWMS 2.0 Rat- Sexing, Handling, Restraint and Ear Ntching RA 2.0 Rat- Sexing, Handling, Restraint and Ear Ntching SWMS 2.1 Rat- Injectin Techniques RA 2.1 Rat- Injectin Techniques SWMS 2.2 Rat Bld cllectin RA 2.2 Rat Bld cllectin SWMS 7.0 Cmpliance - Emergency Cntingency RA 7.0 Cmpliance - Emergency Cntingency SWMS 7.1Cmpliance -Transprtatin RA 7.1 Cmpliance -Transprtatin SWMS Rdent Imprtatin RA 7.2- Rdent Imprtatin SWMS 2.3 Rat Anaesthesia and Analgesia Page 2 19/12/2016

3 Glves Gwn Mask Hair Net She Cvers Persnal Prtective Equipment Required 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 f Equipment Needed Isflurane Inductin Chamber Filler Key Heat Mat Anaesthetic machine Heat mat General Infrmatin All prcedures are t be perfrmed by trained cmpetent staff. Training is available frm senir animal huse staff r Animal Welfare Officer. SWMS 2.3 Rat Anaesthesia and Analgesia Page 3 19/12/2016

4 Evidence f training is available in the Cmpetency Skills Register administered by the Research Services Office. Wmen 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 shuld nt be expsed t isflurane. Anaesthesia and Analgesia Guidelines SWMS 2.3 Rat Anaesthesia and Analgesia Page 4 19/12/2016

5 SWMS 2.3 Rat Anaesthesia and Analgesia Page 5 19/12/2016 Versin 7 Anaesthesia Drug Dse Rute Isflurane 3-5% Inductin Inhalatin 1-4% Maintenance Inhalatin Servflurane 5-8% Inductin Inhalatin 2.5-4% Maintenance Inhalatin Fentanyl µg/kg IP medetmidine 300 µg/kg IP Reverse Medetmidine with atipamezle 1mg/kg IP r SC Reverse Fentanyl with an butrphanl 1-2mg/kg IP r SC Ketamine mg/kg IP acetylprmazine mg/kg IP butrphanl 1.5 mg/kg IP Ketamine mg/kg IP medetmidine 0.5 mg/kg IP Reverse medetmidine with atipamezle 1 mg/kg IP r SC Ketamine mg/kg IP Xylazine 5-10 mg/kg IP Reverse xylazine with atipamezle 1 mg/kg SC Ketamine mg/kg IP diazepam 5 mg/kg IP Pentbarbitne mg/kg (adult) IP 10-20mg/kg (yung) IP Sedatin Acetylprmazine and 2.5 mg/kg IP Butrphanl r 1.5 mg/kg IP Buprenrphine 0.05 mg/kg IP Analgesia Drug- NSAID Dse and frequency Rute Carprfen 5 mg/kg SC hurly Melxicam 1 mg/kg SC 24 hurly Drug- Opiid Dse and frequency Rute Buprenrphine mg/kg SC Buprenrphine 8-12 hurly 0.4 mg/kg in Nutella (give 0.5g t a 250 g rat f 0.2 mg/g nut paste) 24 hurly as per Abelsn et al 2012 PO Butrphanl mg/kg SC 2-4 hurly µ pids may be reversed with butrphanl mg/kg SC Opiid drugs may be reversed IP, IM r with Nalxne 3mg/kg IV Lcal Anaesthesia (Animal Welfare Officer Recmmendatins) Lidcaine dilute t 0.5% d nt exceed 7 mg/kg ttal dse fast nset shrt acting (<1 hur) injectin Bupivacaine dilute t 0.25% d nt exceed 3 mg/kg ttal dse q1-6h infiltratin and epidural and q8-24h plexus r nerve injectin

6 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. Suggested indicatins and prtcls fr researcher/technician training by the AWO and Animal Facility staff: - Rat sedatin fr gavage and mild bleeding prcedures etc: either: - acetylprmazine/butrphanl at 1 t 2.5 mg/kg -1 /1 t1.5 mg/kg -1 i.p r s.c Fr the maximum dse mix 1.25 mls f acetylprmazine ( 10mg/mL) with 0.75 mls f butrphanl (10 mg/ml) and add sterile Nrmal Saline fr injectin up t 5 mls dse at 0.1 ml/ 100 g bdyweight - r, - acetylprmazine/buprenrphine at 1 t 2.5 mg/kg -1 /0.025 t 0.05 mg/kg -1 i.p r s.c Fr the maximum dse mix 1.25 mls f acetylprmazine (10mg/mL) with 0.8 mls f Temgesic (300 mcg/ml) and add sterile Nrmal Saline fr injectin up t 5 mls dse at 0.1 ml/ 100 g bdyweight. Anaesthesia Settings and Setup Check the level f Isflurane and tp up using the methd belw if required. T fill the vapriser yu must have an Isflurane filler key; this is the nly way the vapriser can be filled. 1. 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. 2. 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. 3. 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 tp line in the fill windw. 4. 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 2-5%, a rutine inductin setting is 3% fr mst animals. SWMS 2.3 Rat Anaesthesia and Analgesia Page 6 19/12/2016

7 Maintenance 1-4% Mst animals can be maintained between 1.5 and 3%. Oxygen flw rate (flw meter): Fr 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, prvided the vapriser is less than 5 years f age, has been serviced within the last 18 mnths and is well maintained. 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 t half an hur befre it is required. Ensure it is insulated frm the table tp. 2. Turn n the Oxygen at the wall, if using the Oxygen plumbed int the theatres, r if using an xygen bttle, check that enugh pressure remains in the bttle. Set the flw meter meter t deliver the maximum flw rate pssible t the chamber. Ensure the scavenger is attached and turn n at the right 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. Rats 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 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 3.0 %. An phthalmic intment, such as Lacrilube, shuld be applied t the eyes f the animal and 5 mls f Sterile Nrmal Saline fr Injectin may be 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. SWMS 2.3 Rat Anaesthesia and Analgesia Page 7 19/12/2016

8 9. If recvery takes mre than 15 minutes, such as after injectable anaesthesia, the recvery cage shuld be placed nt a warm mat s that half f its base is in cntact with the mat. This shuld cntinue until the rat is active and fully mbile. Saked fd shuld be prvided in a cntainer n the flr f the cage during a prlnged recvery. 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 Intubatin In situatins which require intubatin, administer either an injectable general anaesthetic, listed in this SWMS, and then place the rat in an anaesthetic inductin chamber with a flw f pure xygen at a minimum rate f 20% f the chamber vlume per minute. Alternatively, induce general anaesthesia using isflurane accrding t the instructins in this SWMS with a pre-xygenatin perid f up t 5 minutes. Once a level f surgical anaesthesia is btained, as determined by an absence f pedal withdraw reflex, fllw the fllwing prcedures t intubate the animal: Variant 1: 1. Remve the animal frm the anaesthetic inductin chamber and place it in a supine psitin, with its head clse t the edge f the prcedure table. 2. Tape the frepaws nt the table with masking tape. 3. Lightly pull dwn n the tp jaw by hking the tp incisr teeth with a lp f suture material. 4. Open the muth and visualise the glttis with a rat laryngscpe. SWMS 2.3 Rat Anaesthesia and Analgesia Page 8 19/12/2016

9 5. Misten the utside f a 12 t 18 gauge, 45 mm, r lnger, intravenus catheter r prprietary endtracheal tube f a similar size. Insert a stiffener, such as a mdified 18 gauge spinal needle. The bevel f the spinal needle shuld be remved and grund t a smth end such that the needle des nt prtrude frm the catheter 6. Pass the end f the stiffened catheter r tube int the glttis and insert until the hub f the catheter engages the muth. 7. Remve the mdified spinal needle and cnfirm crrect placement by nting cndensatin in the catheter. 8. If indicated, cnnect the hub f the catheter t an anaesthetic circuit in rder t maintain anaesthesia r xygenatin f the animal. Variant 2: 1. Remve the animal frm the anaesthetic inductin chamber, hk the upper incisr teeth ver the hrizntal thin crd f the intubatin gantry. 2. Allw the bdy f the rat t be partially suspended by the crd f the gantry. 3. Apprach the animal frm the drsal aspect, pen the muth and visualise the glttis with a rat laryngscpe. 4. Cntinue as fr pint 5 in Variant 1. Mnitring Depth f Anaesthesia NOTE: Rat 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 rat whilst under anaesthesia indicate inadequate depth, and 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 with pure xygen. SWMS 2.3 Rat 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) prcaine/benzathine penicillin (300 mg/ml) at 64 mg/kg by subcutaneus injectin; (iii) enrflxacin (25 mg/ml r less) at 10 mg/kg. Where pssible, based n AWO advice, alternatives t enrflxacin shuld be used; r (iv) 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 2.3 Rat 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 the Animal Welfare Officer r the Animal Facility Manager. SWMS 2.3 Rat Anaesthesia and Analgesia Page 11 19/12/2016