1. The University of New Mexico & The Office of Animal Care and Compliance present Training in Survival Rodent Surgery A learning module developed by the NIH

2. Introduction This module was developed to assist you in becoming more proficient in performing aseptic rodent surgery This module was developed to assist you in becoming more proficient in performing aseptic rodent surgery

3. General Training in Survival Rodent Surgery We all have an ethical responsibility to animals in terms of minimizing pain and distress We all have an ethical responsibility to animals in terms of minimizing pain and distress This can be accomplished, in part, by using proper surgical technique This can be accomplished, in part, by using proper surgical technique The PHS requires that animal care and use is based on the “Guide for the Care and Use of Laboratory Animals” The PHS requires that animal care and use is based on the “Guide for the Care and Use of Laboratory Animals”

4. The following Principles described in the “Guide” apply to rodent surgery Appropriate pre-operative and post-operative care of animals in accordance with established veterinary medical and nursing practices is required Appropriate pre-operative and post-operative care of animals in accordance with established veterinary medical and nursing practices is required All survival surgery will be performed by using aseptic procedures, including sterile gloves, masks, sterile instruments, and aseptic techniques All survival surgery will be performed by using aseptic procedures, including sterile gloves, masks, sterile instruments, and aseptic techniques A dedicated surgical facility is not required for rodents but surgery must be performed using aseptic techniques A dedicated surgical facility is not required for rodents but surgery must be performed using aseptic techniques Research personnel will be appropriately qualified and trained in all procedures Research personnel will be appropriately qualified and trained in all procedures

5. The “Guide” states that it is important for personnel to have appropriate training to ensure that good surgical technique is practiced Good technique includes: Asepsis Asepsis Gentle tissue handling Gentle tissue handling Minimal dissection of tissue Minimal dissection of tissue Appropriate use of instruments Appropriate use of instruments Effective hemostasis Effective hemostasis Correct use of suture materials and patterns Correct use of suture materials and patterns

6. The NIH has established specific recommendations for how survival rodent surgery should be conducted This includes: Preparation of the patient Preparation of the patient Preparation of the surgeon Preparation of the surgeon Sterilization of instruments, supplies, and implanted materials Sterilization of instruments, supplies, and implanted materials Use of operative techniques to reduce the likelihood of infection Use of operative techniques to reduce the likelihood of infection

7. Pre-Surgical Planning begins during the protocol development phase in consultation with the veterinarian This includes: Identification of personnel, their roles and training needs Identification of personnel, their roles and training needs Equipment and supplies required for the procedures planned Equipment and supplies required for the procedures planned The location and nature of the facilities in which the procedures will be conducted The location and nature of the facilities in which the procedures will be conducted Pre- and Post-operative care Pre- and Post-operative care

8. Location, Location, Location You must select the best possible location You must select the best possible location Uncluttered and easily organized and disinfected Uncluttered and easily organized and disinfected Dedicated during the duration of the procedure Dedicated during the duration of the procedure Free of debris and equipment not related to surgery Free of debris and equipment not related to surgery

9. Avoid Locations that are beneath supply ducts to minimize contamination from dust Locations that are beneath supply ducts to minimize contamination from dust High traffic areas such as those near doorways to prevent unnecessary interruptions and creation of air turbulence High traffic areas such as those near doorways to prevent unnecessary interruptions and creation of air turbulence

10. Anesthesia Select anesthetics depending upon the type of surgical procedure Select anesthetics depending upon the type of surgical procedure Anesthetic must be approved in your protocol Anesthetic must be approved in your protocol Most inhalants are administered using a vaporizer Most inhalants are administered using a vaporizer Gas anesthetics require the use of a scavenging system to remove waste gases Gas anesthetics require the use of a scavenging system to remove waste gases Injectable anesthetics are widely used in rodent surgeries Injectable anesthetics are widely used in rodent surgeries If using injectables, weigh each animal and dose according to body weight If using injectables, weigh each animal and dose according to body weight

11. Anesthesia Some anesthetics abolish blink reflex Some anesthetics abolish blink reflex Anesthetized animals should have corneas protected with an ophthalmic ointment Anesthetized animals should have corneas protected with an ophthalmic ointment

12. Monitoring Anesthetized animals must be monitored during the procedure to assure they stay in the proper anesthetic plane Anesthetized animals must be monitored during the procedure to assure they stay in the proper anesthetic plane The anesthetic plane can be assessed by pinching the toes, tail or ears of the animal, or testing the blink reflex The anesthetic plane can be assessed by pinching the toes, tail or ears of the animal, or testing the blink reflex Any reaction of the animal indicates the animal is too light and should be given more anesthetic Any reaction of the animal indicates the animal is too light and should be given more anesthetic

13. Monitoring It is important to inspect the animal and not rely solely on monitoring equipment It is important to inspect the animal and not rely solely on monitoring equipment The color of the mucous membranes and exposed tissues is easy to monitor The color of the mucous membranes and exposed tissues is easy to monitor Bright pink and red as apposed to dusky grey or blue indicates tissue perfusion and oxygenation Bright pink and red as apposed to dusky grey or blue indicates tissue perfusion and oxygenation

14. Monitoring Respiratory pattern and frequency will also give an indication of anesthetic depth Respiratory pattern and frequency will also give an indication of anesthetic depth Core body temperature can also be monitored in rats and mice Core body temperature can also be monitored in rats and mice Pulse oximetry an be used in larger rodents to monitor pulse and oxygenation Pulse oximetry an be used in larger rodents to monitor pulse and oxygenation Electrocardiograms can also be used in larger rodents Electrocardiograms can also be used in larger rodents

15. Complications The most frequent complication of small animal anesthesia is hypothermia The most frequent complication of small animal anesthesia is hypothermia Animals should be provided with a heat source during the pre-, intra- and post-operative periods Animals should be provided with a heat source during the pre-, intra- and post-operative periods Improper heating devices can also be very dangerous Improper heating devices can also be very dangerous Use a thermometer to measure the temperature adjacent to the animal Use a thermometer to measure the temperature adjacent to the animal

16. Electric Heating Pads These are NOT recommended for use with rodents as they have varying temperatures across the surface These are NOT recommended for use with rodents as they have varying temperatures across the surface Whew! Is it me, or is it hot?

17. Slide Warmers Slide warmers can be used as a heat source during recovery Slide warmers can be used as a heat source during recovery Animals must be watched closely and placed in recovery cage as soon as they begin to stir Animals must be watched closely and placed in recovery cage as soon as they begin to stir The safest device is a circulating hot water blanket The safest device is a circulating hot water blanket All heating devices should be covered with a towel so the animal doesn’t come in contact with the heated surface All heating devices should be covered with a towel so the animal doesn’t come in contact with the heated surface

18. Instruments A simple peel pack can contain small numbers of small to medium sized instruments A simple peel pack can contain small numbers of small to medium sized instruments

19. Instruments A complex pack consists of overlapping cloth or paper drapes folded together and sealed with autoclave tape A complex pack consists of overlapping cloth or paper drapes folded together and sealed with autoclave tape

20. Multiple Rodent Surgeries When performing multiple rodent surgeries have staging areas for the different steps in the procedure When performing multiple rodent surgeries have staging areas for the different steps in the procedure Animals waiting for surgery should be kept at a visual and olfactory distance from those undergoing surgery Animals waiting for surgery should be kept at a visual and olfactory distance from those undergoing surgery Yuck! What’s that smell?

21. Space Constraints If space necessitates a single location for prep and surgery, the bench towel used during prep should be replaced for the surgery If space necessitates a single location for prep and surgery, the bench towel used during prep should be replaced for the surgery Any open surgery pack must be covered to prevent contamination with hair Any open surgery pack must be covered to prevent contamination with hair Hey! I don’t shed.

22. Prepare The Area Sanitize the area you have selected for performing the surgery with an appropriate disinfectant Sanitize the area you have selected for performing the surgery with an appropriate disinfectant Disinfectants include: Clidox, Alcide, or Nolvasan Disinfectants include: Clidox, Alcide, or Nolvasan A dedicated surgical site is not required for rodent surgery A dedicated surgical site is not required for rodent surgery

23. Step 1 Once everything is pre-selected and organized, it is time to begin Once everything is pre-selected and organized, it is time to begin Start by anesthetizing the animal and doing the surgical prep Start by anesthetizing the animal and doing the surgical prep Hair must be removed from the surgical site Hair must be removed from the surgical site Electric clippers are commonly used Electric clippers are commonly used

24. Hair Removal An easy alternative to clipping is hair plucking An easy alternative to clipping is hair plucking Hair follicles in mice are usually in telogen or resting phase and hair can be removed without injury Hair follicles in mice are usually in telogen or resting phase and hair can be removed without injury

25. Surgical Prep Standard surgical prep consists of alternating scrubs of an iodophor and 70% alcohol Standard surgical prep consists of alternating scrubs of an iodophor and 70% alcohol

26. Scrub Procedure Using a gauze sponge or cotton tipped applicator, cleansing should be done in a circular motion Using a gauze sponge or cotton tipped applicator, cleansing should be done in a circular motion Begin at the center of the shaved area and work toward the periphery Begin at the center of the shaved area and work toward the periphery Never go back to the center with the same sponge Never go back to the center with the same sponge Scrubs should be alternated between an iodophor scrub and alcohol, ending with the iodophor Scrubs should be alternated between an iodophor scrub and alcohol, ending with the iodophor

27. Surgical Pack Once the animal is prepped you must open the surgical pack Once the animal is prepped you must open the surgical pack This is to be done BEFORE donning surgical gloves This is to be done BEFORE donning surgical gloves Make sure the sterilization indicator has turned the appropriate color (usually autoclave tape – brown stripes) Make sure the sterilization indicator has turned the appropriate color (usually autoclave tape – brown stripes)

28. Peel Packs Peel packs are opened in a manner that preserves the sterility of the inside surface Peel packs are opened in a manner that preserves the sterility of the inside surface Do not touch the inside surface as it can be used as a sterile surgical field on which to keep the instruments Do not touch the inside surface as it can be used as a sterile surgical field on which to keep the instruments

29. Complex Packs Complex packs are opened in such a way as to keep the inside surface of the wrapping sterile so that it can be used as a sterile field Complex packs are opened in such a way as to keep the inside surface of the wrapping sterile so that it can be used as a sterile field

30. Liquid Sterilants If using cold sterilant solutions make sure instruments are exposed for the proper length of time and expiration dates of solutions is observed If using cold sterilant solutions make sure instruments are exposed for the proper length of time and expiration dates of solutions is observed Instruments must be removed from solution and rinsed with sterile water, saline, or alcohol Instruments must be removed from solution and rinsed with sterile water, saline, or alcohol Rinsed instruments must be placed on a sterile field Rinsed instruments must be placed on a sterile field

31. Hot Bead Sterilizers This method sterilizes only the tips of the instruments This method sterilizes only the tips of the instruments Beads must be pre-heated to the recommended temperature and the instruments exposed for the recommended time Beads must be pre-heated to the recommended temperature and the instruments exposed for the recommended time Gross debris must be removed from the instrument prior to sterilization Gross debris must be removed from the instrument prior to sterilization Instrument must be allowed to cool before touching tissues Instrument must be allowed to cool before touching tissues Best used for sterilizing instruments between surgeries Best used for sterilizing instruments between surgeries

32. Delicate Instruments Delicate instruments, materials for implantation or items that otherwise may melt or become damaged when heated can be sterilized using ethylene oxide Delicate instruments, materials for implantation or items that otherwise may melt or become damaged when heated can be sterilized using ethylene oxide The packs must be sufficiently aerated to prevent toxic side effects from residual gas The packs must be sufficiently aerated to prevent toxic side effects from residual gas This may require 24 to 72 hours This may require 24 to 72 hours

33. Notes: “Flash” dry heat sterilizers are another alternative for sterilizing instruments – remember to let them cool before touching tissues “Flash” dry heat sterilizers are another alternative for sterilizing instruments – remember to let them cool before touching tissues Remember – alcohol provides disinfection not sterilization Remember – alcohol provides disinfection not sterilization

34. Instrument Packs Once packs are opened all other sterile equipment must be placed on the sterile field Once packs are opened all other sterile equipment must be placed on the sterile field These items must be opened in a way as to prevent contamination of the item or the surgical pack These items must be opened in a way as to prevent contamination of the item or the surgical pack

35. Surgical Attire Cap Cap Mask Mask Clean lab coat Clean lab coat Wash hands with antibacterial soap and don sterile gloves Wash hands with antibacterial soap and don sterile gloves

36. Donning Surgical Gloves Open the package of gloves observing sterile technique Open the package of gloves observing sterile technique Remember, the inside of the package is STERILE – exam gloves are not the same as sterile gloves Remember, the inside of the package is STERILE – exam gloves are not the same as sterile gloves

37. Donning Surgical Gloves

38. Donning Surgical Gloves - Procedure It is important to don the gloves in such a way that prevents contamination of the outer surface of the gloves It is important to don the gloves in such a way that prevents contamination of the outer surface of the gloves One glove is lifted from the opened glove package by its turned down cuff One glove is lifted from the opened glove package by its turned down cuff The glove is pulled on the hand with a rotating motion The glove is pulled on the hand with a rotating motion Place the gloved fingers beneath the cuff of the other glove Place the gloved fingers beneath the cuff of the other glove With the gloved fingers under the cuff, the glove is placed on the ungloved hand With the gloved fingers under the cuff, the glove is placed on the ungloved hand The folded cuff protects the gloved hand from contamination The folded cuff protects the gloved hand from contamination It is pulled over the cuff of the lab coat following insertion of the hand It is pulled over the cuff of the lab coat following insertion of the hand The fingers are then slipped under the cuff of the first glove to pull it over the lab coat cuff The fingers are then slipped under the cuff of the first glove to pull it over the lab coat cuff

39. The Surgical Site - Draping The most common drape is the paper drape The most common drape is the paper drape It may be precut or one in which you must cut a hole It may be precut or one in which you must cut a hole A disadvantage to paper drapes is that they usually cover the animal making monitoring difficult A disadvantage to paper drapes is that they usually cover the animal making monitoring difficult Plastic drapes offer the advantage of more visibility Plastic drapes offer the advantage of more visibility Sterile gauze sponges can also be used for drapes Sterile gauze sponges can also be used for drapes

40. Organize Organize the instruments in your surgical pack Organize the instruments in your surgical pack Point all tips in one direction Point all tips in one direction It is helpful to place them in the order used It is helpful to place them in the order used Between surgeries cover the tips of the instruments with sterile gauze Between surgeries cover the tips of the instruments with sterile gauze Note that the space between the pack and the draped animal is not sterile, do not lay instruments in this space Note that the space between the pack and the draped animal is not sterile, do not lay instruments in this space

41. During Surgery - Drapes Be careful not to get paper or cloth drapes wet Be careful not to get paper or cloth drapes wet Wet material acts as a wick to pull bacteria through from the non-sterile surface below Wet material acts as a wick to pull bacteria through from the non-sterile surface below When this happens instruments should be considered contaminated When this happens instruments should be considered contaminated

42. Suture Selection and type should be based upon the type of surgery and species of animal Selection and type should be based upon the type of surgery and species of animal This should be done before hand after consultation with the veterinarian This should be done before hand after consultation with the veterinarian I don’t want to talk to the vet. Suture self

43. Sutures and More Sutures For small animals a 3-0 suture thickness or smaller is best For small animals a 3-0 suture thickness or smaller is best Cutting and reverse cutting needles have sharp edges are best for skin suturing Cutting and reverse cutting needles have sharp edges are best for skin suturing Non-cutting, taper or round needles are used for suturing easily torn tissues such as peritoneum, muscle or intestine Non-cutting, taper or round needles are used for suturing easily torn tissues such as peritoneum, muscle or intestine

44. … and More Sutures Ligation of vessels or suturing tissues other than skin requires an absorbable material such as Vicryl, Dexon, PDS, Maxon, or chromic gut Ligation of vessels or suturing tissues other than skin requires an absorbable material such as Vicryl, Dexon, PDS, Maxon, or chromic gut Skin closures require a non-absorbable suture such as Prolene or nylon Skin closures require a non-absorbable suture such as Prolene or nylon Stainless steel wound clips or staples can also be used Stainless steel wound clips or staples can also be used Most rodents gnaw at externalized sutures so a buried suture line or wound clips are recommended Most rodents gnaw at externalized sutures so a buried suture line or wound clips are recommended

45. … and Finally Cyanoacrylate surgical adhesives such as Vetbond or Nexaband may be used to close incisions or the area between sutures Cyanoacrylate surgical adhesives such as Vetbond or Nexaband may be used to close incisions or the area between sutures Silk is a non-absorbable suture material that can cause tissue reactions and may wick microorganisms into the wound Silk is a non-absorbable suture material that can cause tissue reactions and may wick microorganisms into the wound Silk is best used for cardiovascular procedures only Silk is best used for cardiovascular procedures only

46. Antibiotics The “Guide” states that the application of prophylactic antibiotics is not a substitute for the practice of proper aseptic surgery The “Guide” states that the application of prophylactic antibiotics is not a substitute for the practice of proper aseptic surgery If they must be used, you must choose an appropriate antibiotic that is given for the correct length of time and dose If they must be used, you must choose an appropriate antibiotic that is given for the correct length of time and dose In guinea pigs and hamsters an inappropriate antibiotic can cause fatalities In guinea pigs and hamsters an inappropriate antibiotic can cause fatalities

47. Recovery Keep the recovering patient warm Keep the recovering patient warm Do not lay recovering animals directly on the bedding – they may aspirate and asphyxiate Do not lay recovering animals directly on the bedding – they may aspirate and asphyxiate

48. Analgesia Part of the surgical planning should include intra-operative and post-operative analgesia Part of the surgical planning should include intra-operative and post-operative analgesia Oh Yeah! I likes dem drugs!

49. Fluids Recovery from anesthesia can be aided by the administration of warmed fluids Recovery from anesthesia can be aided by the administration of warmed fluids Fluids should be given subcutaneously or intra-peritoneally Fluids should be given subcutaneously or intra-peritoneally Consult the veterinarian for appropriate volumes and routes of administration Consult the veterinarian for appropriate volumes and routes of administration Funny, I gotta pee.

50. Neonates Neonates or animals recovering from prolonged surgeries can suffer from hypoglycemia Neonates or animals recovering from prolonged surgeries can suffer from hypoglycemia These animals can benefit from administration of oral glucose These animals can benefit from administration of oral glucose Glucose should never be given SQ or IP Glucose should never be given SQ or IP

51. Return to the Cage Animals can be returned to their holding area once they are awake and appear to be making a normal recovery Animals can be returned to their holding area once they are awake and appear to be making a normal recovery Be sure to mark the cage card with the surgical procedure and the date Be sure to mark the cage card with the surgical procedure and the date Post-operative care does not end with the return of the animal to its home environment Post-operative care does not end with the return of the animal to its home environment

52. After the Surgery Animals must be monitored for several days after the surgical procedure Animals must be monitored for several days after the surgical procedure Daily weighing is a sensitive method of monitoring the animal Daily weighing is a sensitive method of monitoring the animal While subtle changes in activity or appetite may not be observed, changes in weight will be quickly detected While subtle changes in activity or appetite may not be observed, changes in weight will be quickly detected Some analgesics depress appetite and must be differentiated from that which occurs if an animal is not feeling well Some analgesics depress appetite and must be differentiated from that which occurs if an animal is not feeling well

53. Continued Monitoring Animals must be monitored for the continued need for analgesics Animals must be monitored for the continued need for analgesics This assessment should be made at least twice daily in the first few days post-op This assessment should be made at least twice daily in the first few days post-op Supplying a softer, more palatable, easily accessible diet may encourage the animal to eat Supplying a softer, more palatable, easily accessible diet may encourage the animal to eat Hydration can be checked by “tenting” the animal’s skin Hydration can be checked by “tenting” the animal’s skin In a well hydrated animal the skin will quickly fall back in place when released In a well hydrated animal the skin will quickly fall back in place when released In a dehydrated animal the skin will be slow to return to its original place In a dehydrated animal the skin will be slow to return to its original place

54. Wound Closures Wound closures must be removed at 10 to 14 days post-op Wound closures must be removed at 10 to 14 days post-op Suture scissors or staple removers must be used Suture scissors or staple removers must be used

55. This Concludes Module 2 – Training in Survival Rodent Surgery Please download the exam, complete it, then e- mail it to KMirowsky@salud.unm.edu Please download the exam, complete it, then e- mail it to KMirowsky@salud.unm.eduKMirowsky@salud.unm.edu