STANDARD OPERATING PROCEDURE:

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1 STANDARD OPERATING PROCEDURE: Perfrming a rat dissectin Nte: T be undertaken nly by trained persnnel in cnjunctin with a current Safety Data Sheet (SDS) and site-specific risk assessment. 1. Intrductin A rat dissectin is cnducted t explre the internal structure and functin f basic mammalian anatmy. The purpse f this exercise is t explre the alimentary canal f the rat and bserve the different parts f the digestive system. By lking at the length f the digestive system f the rat, infrmatin abut their dietary pattern can be determined, and cmparisns made between the rat and ther mammals such as mnivres, carnivres and herbivres. Because the rat is a placental mammal and an mnivre it has a bdy structure very similar t that f humans. Prir t cnducting a rat dissectin, teachers shuld ensure that they are able t meet the requirements f the Schls Animal Ethics Cmmittee (SAEC) in their jurisdictin. It is recmmended that they cnsider the educatinal bjectives fr this activity, explre the ethical cnsideratins with students and aim t reduce the ttal number f rats required fr this activity. Students shuld nt be frced t participate in a dissectin and alternative activities such as vides and virtual dissectins can be used fr these students instead, as well as t supplement the actual activity. Rats that have been humanely euthanised and are disease (r infectin) free shuld be surced frm ethical and licenced suppliers. They can be supplied as freshly euthanised, frzen r preserved specimens. If using preserved rats, btain and read the safety data sheet (SDS) frm the supplier and prepare a site-specific risk assessment. If freshly euthanised r recently defrsted rats are being used, and the dissectin is interrupted, the rats can be kept fr a shrt time (n lnger than 24 hurs) in the cldest part f the labratry fridge r placed in a freezer. There is ptential fr bacterial grwth. 2. Cntext These instructins are fr the use f experienced science teachers, technicians and students under clse supervisin. When planning a class dissectin activity it is best t discuss befrehand the type f dissectin t be undertaken, and warn f the pssibility that there may be sme bld and durs present during the dissectin. Demnstrating the dissectin t students befre they begin is helpful, nt nly fr crrect prcedure but allws them t adjust t the appearance f the material, and any bld that may be present after dissectin material has been washed. Let students knw they dn t have t participate in the dissectin and can be excused frm the class. Alternative arrangements can be made fr students wh dn t wish t participate, by giving them wrksheets t cmplete and relcating them t a private study area. Page 1 f 12

2 3. Safety ntes Fainting: signs and symptms: Fainting may ccur during this type f activity. Please read the first aid infrmatin in sectin 7 befre cnducting the dissectin. Fainting is caused by a sudden drp in bld pressure. Cmmn causes include heat, pain r distress and the sight f bld. The pssible symptms include the fllwing. Dizziness Light-headedness A pale face Perspiratin Heightened anxiety and restlessness Nausea Cllapse Uncnsciusness, fr a few secnds Full recvery after a few minutes i Handling specimens: If using preserved rats, it is imprtant t take nte f the preservative slutin that the rats are in and the recmmended precautins. Rinse the preserved rats under running water immediately upn remval frm the preservative slutin. Wrk in a well-ventilated area. It is recmmended that peple wearing cntact lenses shuld nt dissect rats that are in preservative slutin. The fumes frm the slutin can penetrate between the eye and cntact lens causing irritatin t eyes. It is recmmended t wear prescriptin glasses instead with safety glasses ver them OR prescriptin safety glasses. Cnsider issues such as allergies and chemical sensitivities frm handling freshly euthanised r recently defrsted r preserved rats. Be aware f pssible micrbial aersls and unpleasant durs released frm freshly euthanised r recently defrsted rats if the stmach/intestines are accidently cut. If using frzen rats, defrst vernight in a refrigeratr befrehand and use within 24 hurs. Gd hygiene practices shuld be bserved at all times: Keep hands away frm the muth, nse, eyes and face during and after dissectin and wash hands immediately after handling dissectin material. Safety with scalpels: Ensure students demnstrate respnsible behaviur while using scalpels. Only staff shuld carefully attach and remve scalpel blades. The scalpel blade size and handle must be cmpatible, e.g. number 4 handle with number 23 blade. Keep the blade in the fil wrapper and attach t the handle with the sharp side f the blade pinting away frm the bdy. Blades shuld be remved carefully using pliers, frceps r a cmmercial blade remver. An alternative is t use dispsable scalpels. Page 2 f 12

3 4. Regulatins, licences and permits Sme SAECs may require permissin well in advance f the activity being cnducted. In sme jurisdictins, all dissectins need t be reprted t the SAEC. Science ASSIST has develped an infrmatin sheet with links t bilgical safety and jurisdictinal SAECs, see AIS: Links Bilgical sciences safety Rats that have been humanely euthanised and are disease (r infectin) free shuld be surced frm ethical and licenced suppliers. 5. Equipment PPE Lab cat/aprn (it is recmmended t use plastic dispsable aprns), safety glasses and glves. Scalpels Scissrs Frceps Prbes Dissecting pins/needles Dissecting bards cvered with newspaper. Glazed ceramic tiles, nyln plastic bards, vinyl dissecting pads r dispsable fam trays are suitable. String Paper twel Disinfectant hspital grade, general purpse disinfectant (the label n the frnt f the pack must state hspital grade, which is a general purpse hard-surface disinfectant which will kill micr-rganisms). 70% v/v ethanl Page 3 f 12

4 6. Operating prcedure Preparatin If any bld is assciated with the rats rinse them in cld running water. Prepare disinfectant slutin accrding t manufacturer s instructins. Place disinfectant in a cntainer ready fr instruments t be placed at the end f the dissectin. Ensure students have apprpriate PPE. Distribute the instruments t students. Scalpels, scissrs, frceps, dissecting pins and prbes shuld be cunted ut, and cunted in when returned. Examining and dissecting the rat 1. First identify all the external structures visible n the head, thrax and abdmen f the rat. See Figure 1a. 2. Observe the muth cavity and teeth f the rat. 3. Place the rat n the dissecting tray n its back. String is used t fasten the limbs f the rat back. Tie string firmly arund ne wrist, pass the string under the tray and tie it firmly t the ther wrist. Repeat with the hind limbs. See Figure 1b. 4. Pick up the skin in the middle f the ventral surface using frceps. Cut (see incisin lines in figure 1b) the skin and underlying muscles with the blunt end f the scissrs pinting dwnwards. Cut twards t the bttm f the ribs and then in the ther directin t the genital penings. Cut acrss the bttm f the ribs, and then cut ut t bth the hind legs. 5. Peel back the flap f skin frm the underlying muscle and ntice the cnnective tissue between them. Use the dissecting pins t pin ut the skin f the rat t the dissecting bard. See Figure Open up the abdminal cavity by lifting the abdminal wall with frceps, make an incisin with the scissrs and fllw the same prcedure as in step 4 abve. Be careful that yu d nt damage any f the abdminal rgans by making the incisin t deep. 7. Peel back the abdminal wall and pin ut revealing the abdminal rgans. See Figure 3. Examine the digestive rgans in situ and identify all the structures in place. Ntice the scattered cream clured fat depsits. Nte the size f the liver in prprtin t the abdmen. 8. Remve the digestive system by cutting the esphagus abve its cnnectin with the stmach and by cutting the rectum. Ntice the way the small intestine and large intestines are twisted ver ne anther. See Figure Using yur fingers, carefully pull apart the dudenum and cln frm the cnnective tissue by gently tearing the mesentery and carefully stretch ut the alimentary canal. Measure the length f the alimentary canal and the rat s bdy cmparing the rati f these figures. See Figure Nte the size f the liver and caecum in relatin t the rat s bdy. (Operating prcedure cnt.) Page 4 f 12

5 11. Remve the liver carefully and cmpare the clur f the spleen and liver. Nte there is n gall bladder in the rat. 12. Observe the expsed kidneys and assciated urinary rgans, and the reprductive system left behind in the abdminal cavity. See Figure If time allws fr further investigatin, pen the thracic (chest) cavity by making a midventral cut using blunt end f scissrs cutting frward thrugh the diaphragm muscle and breast bne up t just under the chin f the rat. 14. The diaphragm, which separates the thracic frm the abdminal cavity, can be cut away frm the rib cage. 15. Cut away the ribs t reveal the heart surrunded by a thin membrane called the pericardium, the whitish thymus gland that lies directly ver the upper part f the heart and the lungs that lay either side f the heart. See Figure Remve the heart and thymus gland t lcate the trachea (wind pipe) identifiable by its rings f cartilage. The esphagus lies just underneath the trachea. Mre detailed infrmatin n a rat dissectin can be fund n the fllwing websites belw. Digestive System. Rat digestive system Rat dissectin, Clean up: Make sure all instruments are returned. All parts f the rat, as well as the dispsable fam tray (if used), must be wrapped in newspaper and placed in a dedicated plastic garbage bag alng with glves and dispsable aprns (if used). When all waste material is cllected, duble bag fr dispsal. Freeze material if unable t dispse f immediately. If bld is present n dissecting bards, scissrs, frceps, prbes, dissecting pins and scalpels they be immediately saked in disinfectant. Otherwise wash equipment in ht sapy water and rinse r place in dishwasher t minimize handling. After washing, dissecting instruments can be saked in 70% v/v ethanl fr 20 minutes as an ptinal additinal disinfectant and t avid rusting. Dry all equipment thrughly. Disinfect wrkplace and wash hands thrughly. 7. Truble shting/emergencies If fainting ccurs: If students start t feel faint, dizzy r nauseus during the dissectin lie them dwn (if pssible) and elevate their feet. They can get up slwly after ten minutes. Sending them utside fr sme fresh air can als help. If they dn t recver quickly, always seek urgent medical attentin. D nt sit the patient n a chair with head between knees ii Page 5 f 12

6 Any health cncerns shuld be referred t the schl first aid fficer fr assessment, accmpanied by the relevant latest SDS if applicable. Fllw yur schl s accident and incident plicy and reprting prcedures. First Aid: See latest SDS f any chemicals used fr mre detailed infrmatin. If swallwed: D nt induce vmiting. Rinse muth with water, and then give water t drink. Seek urgent medical attentin. If in eyes: D nt induce vmiting. Rinse muth with water, and then give water t drink. Seek urgent medical attentin. If n skin/clthes: If spilt n skin r clthes quickly wipe ff with a dry clth t absrb as much liquid as pssible. Remve cntaminated clthes and drench the area with excess water under a safety shwer. Seek medical attentin. If inhaled: Remve t fresh air and seek medical attentin if symptms persist. Fr further advice cntact the Pisns Infrmatin Centre n Care shuld be taken with sharps such as scalpel blades and scissrs. See safety ntes if it is necessary t remve brken r used scalpel blades. Stre all dissecting instruments securely. First aid: cuts and laceratins shuld be washed under running water, then patted dry and cvered with a clean paper twel r tissue in the first instance and referred t the schl first aid fficer fr assessment. 8. Waste dispsal Used and damaged scalpel blades must be placed in an apprved sharps cntainer after use. Bilgical material must be wrapped in newspaper, placed in a duble plastic garbage bag and sealed fr immediate dispsal in the industrial bins. Specimens preserved in Carlina s Perfect Slutin are nt subject t hazardus materials regulatins r dispsal restrictins in Australia. Refer t SDS and treat as abve fr bilgical material. 9. Related material Risk Assessment. Manufacturer s Safety Data Sheet fr disinfectant Manufacturer s Safety Data Sheet fr preserved specimens References: i Fainting, Better Health Channel website, State Gvernment f Victria: (August 2014) ii St Jhn Ambulance Australia Australian First Aid. Bartn, ACT Andrews, C; Naidu, Satya; Laidler, Greg Active science: skills and experiments: bk 2. Oxfrd University Press: Suth Melburne, Vic. Page 6 f 12

7 Animal use decisins, WA Department f Educatin website, (September 2014) Chemwatch Gld Safety Data Sheet: Hspital grade disinfectant. Chemwatch website (Subscriptin required. Accessed January 2016). Digestive System. Rat digestive system, Wikispaces, (Accessed January 2016) Safety nte Preserved specimens, Suthern Bilgical website, eservedspecimens.pdf (Accessed February 2016) Specimens in Carlina s Perfect Slutin, Material Safety Data Sheet, Carlina Bilgical Supply Cmpany, Suthern Bilgical website, nspecimens.pdf (April 2011) Page 7 f 12

8 Figures Muth Whiskers Chin Neck Chest Nse Incisrs Eyes (bscured) Ear Fre arm Fre paw Skin and fur Abdmen Hind leg Genital pening/penis Anus Scrtal sac Hind feet Tail Figure 1a: The external structures visible n the head, thrax (chest) and abdmen including incisin lines numbered 1, 2 and 3. Figure 1b: The dtted lines shw the incisin lines fr dissectin f rat. (Image by K. Szalai, 2016) Figure 2: The abdminal wall expsed. (Image by K. Szalai, 2016) Page 8 f 12

9 Breastbne Diaphragm Liver Intestines and cnnective tissue Large intestine Figure 3: The abdminal rgans f the rat in situ. (Image by K. Szalai, 2016) Male reprductive rgans Bladder Anus Liver Stmach Figure 4: Abdminal rgans remved. (Image by K. Szalai, 2016) Spleen Cnnective tissue (mesentery) Small intestine Large intestine Rectum Page 9 f 12

10 Liver Stmach Spleen Pancreas Dudenum Small intestine Figure 5: The alimentary canal f the rat with the cnnective tissue remved. (Image by K. Szalai, 2016) Large intestine Appendix Caecum Cln Rectum Anus Kidney Prtective fat Seminal Vesicle Prtective fat layer Bladder Testes Thyrid gland and trachea Thymus gland Lungs. Right lbe. Prbe pints t left lbe. Heart Diaphragm Penis Figure 6: Male Uringenital system (Image by K. Szalai, 2016) Figure 7: Thrax and neck. (Image by K. Szalai, 2016) Page 10 f 12

11 Glssary Abdminal wall the lining that enclses the abdminal cavity, cnsisting mstly f muscle. Alimentary canal the tubular passage between the muth and the anus. Anus the last part f the large intestine thrugh which the faeces are excreted. Appendix a small tube with nly ne pening, near the beginning f the large intestine. Bladder the rgan where urine is stred in the bdy. Bld liquid that circulates in the bld vessels f many animals. It cntains plasma, red and white bld cells, platelets, disslved gases, nutrients, hrmnes and salts. Breast bne the lng flat bne shaped in the middle f the chest, frming the frnt f the rib cage. Caecum large rgan where the small intestine jins the large intestine. Carnivre an animal that eats nly meat. Cartilage flexible cnnective tissue in animals, including the jints between bnes, the rib cage, the ear, the nse, the brnchial tubes and the intervertebral discs. Cln the sectin f the large intestine that runs frm the caecum t the rectum and absrbs water frm faeces. Cnnective tissue animal tissue that supprts, cnnects, and surrunds rgans and ther bdy parts. Diaphragm the muscle that runs acrss the base f the chest cavity and separates the thrax and the abdmen. It causes the lungs t expand and cntract during breathing. Digestin the prcess f cnverting large, cmplex rganic mlecules t smaller, simpler nes that can pass thrugh cell membranes. Digestive system the parts f the bdy where fd is digested. Dudenum the tp part f the small intestine, where bile and pancreatic juice enter the digestive system. Faeces undigested fd and bacteria that is stred in the rectum and expelled thrugh the anus. Gall Bladder small rgan that is lcated beneath the liver and drains bile int the dudenum primarily t break dwn fat during digestin. Heart muscular rgan that pumps bld arund the bdy. Herbivre an animal that eats nly plants. Incisrs the tw frnt teeth in each jaw f a mammal used fr cutting fd. In situ in its riginal place. Kidneys the majr excretry rgans that filter the bld, and maintain the cncentratin f water and salts in the bdy. They frm urine in which excess salts, water urea and waste prducts are remved frm the bdy. Page 11 f 12

12 Large intestine the end sectin f the alimentary canal reaching frm ileum t anus and cnsisting f the caecum, cln, and rectum. Its functin is t extract water and frm faeces. Liver a large rgan in mammals that cntrls the level f nutrients and txic substances in the bld. Has a digestive functin, secretes bile, filters bld, and takes part in many metablic functins such as the cnversin f sugars int glycgen. Lungs inflatable rgans used t breathe in xygen and breathe ut carbn dixide. Mammal class f warm-blded vertebrate animals. Mechanical digestin the breaking dwn f large pieces f fd int smaller pieces. Mesentery a supprtive membrane surrunding and giving structure t the inner rgans. Oesphagus the passage dwn which fd mves between the thrat and the stmach. Omnivre an animal that eats bth plants and animals. Pancreas an rgan in the bdy that prduces pancreatic juice that helps t digest fd. Penis male sex rgan used t transfer semen and expel urine frm the bdy. Pericardium membrane that frms a sac surrunding the heart and attached prtins f the main bld vessels. Placental mammal the embry develps in the maternal uterus attached t the tissues f the placenta. Rectum the last part f the large intestine where faeces is stred. Scrtal sac the sac f skin that hlds the testicles. Seminal vesicle the gland that secretes seminal fluid in semen. Small intestine the part f the intestine between the stmach and the large intestine, cnsisting f the dudenum, jejunum, and ileum, where digestin f fd and mst absrptin f nutrients take place. Spleen rgan in the left upper abdmen f humans and ther vertebrates that helps t destry ld red bld cells, frm lymphcytes, and stre bld. Stmach a hllw muscular rgan where mechanical and chemical digestin takes place. Testes male sex rgans that prduce sperm. Thracic relates t the chest and cavity f an animal. Thrax the part f the bdy between the neck and abdmen, enclsed by the ribs and cntaining lungs, heart and diaphragm. Thymus the rgan, lcated at the base f the neck, and is invlved in develpment f cells f the immune system. Thyrid gland endcrine gland lcated in the neck f human beings and ther vertebrate animals that secretes the hrmnes respnsible fr cntrlling metablism and grwth. Trachea a tube thrugh which air enters the lungs frm the nse. Ventral surface the surface f the abdmen r lwer bdy. Page 12 f 12

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