Comments & suggestions  should be directed to jroese@lssu.edu Visit my website ANIMAL CARE AND USE Arrangements for Acquisition and Housing Acquisition of animals is the responsibility of the PI/CI.  All animals must be housed within IACUC approved facilities.  Arrangements for housing must be made before any animals are acquired.  Animals may not be purchased or otherwise acquired until a fully approved protocol is on file.  If wild animals are to be used, arrangements for any necessary quarantine must be made through the IACUC before animals are acquired.  The PI/CI is responsible for determining if permits (such as from the U.S. Fish and Wildlife Service or the Michigan Department of Natural Resources) are required and any necessary permits must be obtained before animals are acquired.  The IACUC can advise investigators whether permits are needed. Investigators requiring special care, equipment, or supplies for their animals or exemptions from standard animal care procedures must inform the IACUC in writing so that appropriate arrangements can be made.  Users must notify the IACUC if their animals will be exposed to materials or procedures which may be hazardous to personnel. Animal Identification and Record Keeping The Animal Welfare Act and the Guide require appropriate identification of animals and maintenance of animal records. Accepted methods of animal identification include room, rack and cage cards; collars and bands; ear notches and tags; implantable microchips; tattoos; and freeze brands.  Toe clipping is only acceptable for rodents and lower vertebrates when other methods of identification cannot be used. Investigators are responsible for maintaining appropriate records on all animals including rodents, bats, birds and ectotherms.  Cage or rack cards should indicate, at a minimum, the source of the animal and the name and telephone number of the responsible investigator, an IACUC approval number and its expiration date.  Written records of procedures, drug use, illnesses, injuries, and date of death, euthanasia, or disposition should be noted. Animal Husbandry (top of page) The animal care facilities are located in the basement of the Crawford Hall of Science (Suite 119).  Animals may not be housed in other locations for periods of longer than 12 hours without prior approval of the study area by the IACUC.  Regardless of where animals are housed the PI/CI has responsibility for the procurement of animal cages, feed, bedding, and other supplies. They must also provide, and are responsible for, the daily husbandry and the general health of the laboratory animals under their care. Caging Cage design and construction material can influence study results.  Galvanized caging material or rubber bottle stoppers can serve as a source of trace minerals which could affect the results of studies where the level of these compounds is being controlled.  Other important considerations include whether contact bedding can be used or if animals must be housed on a wire floor.  Physiological studies may require the use of a metabolic cage, or observation studies may require the use of clear rather than opaque caging.  The behavioral characteristics of the animal will also dictate the type of cage design used.  For example, some animals require perches, nesting boxes or hiding places.  Reproductive needs may require specific caging features.  In some species females must have a method of escape from an over aggressive male.  Many neonates have inadequate homeothermic mechanisms and will become hypothermic if not protected by contact bedding or nesting material placed in the cages. Consideration should also be given to cage size.  There are specific cage size requirements set forth in the Guide and by the Animal Welfare Act.  Cage size requirements depend upon the species, weight or size of the animal(s), number of animals in the cage and breeding status.  In addition to the floor space requirements the behavioral characteristics of the species, strain, and sex must be considered when group- housing animals.  For social animals, individual housing may cause stress. Among social animals, the formation of new groups can result in fatal trauma from fighting.  Male mice will often fight when group housed, whereas male rats typically do not.  Even in docile animals, overcrowding can lead to fighting, cannibalism, and stress.  Breeding activity can be significantly modified by group housing arrangements. For example, group-housing female mice can lead to anestrus with subsequent estrus synchronization with the introduction of a male mouse. The PI/CI is responsible for the selection of appropriate cages for laboratory animals and for ensuring that housing conforms to Guide standards and Animal Welfare Act requirements.  They are also responsible for maintaining sanitary conditions and keeping cages in good repair.   Exceptions to Guide standards must be justified on the basis of experimental or species requirements. Temperature and Humidity (top of page) The temperature and humidity in the animal room (macroenvironment) should be monitored and maintained within acceptable limits (30-70%). The temperature and humidity in the microenvironment is more difficult to monitor and control.  Variations in temperature and humidity are influenced by such factors as filter tops, hanging wire or solid bottom caging, population density, animal activity level, cage location, and temperature and humidity in the animal room itself. Variations in temperature and humidity can have a variety of effects.  For example, low humidity has been associated with a rodent lesion called ring tail which is characterized by annular constrictions and can result in loss of the tail.  More subtle temperature and humidity effects include: altered drug metabolism, increased disease susceptibility and decreased reproductive efficiency. These examples serve to illustrate the need for controlled temperature and humidity in the animals' micro and macroenvironment and the vital role it plays in the generation of consistent, reliable data.  Principle Investigators and Instructors are responsible for monitoring and maintaining appropriate temperature and humidity in animal facilities. Ventilation Ventilation in animal rooms can have significant impact on the health status of the occupants.  Excessive odor is often the first indication of a ventilation problem in an animal room.  Investigators should be aware, however, that the concentration of waste gases at cage level is usually higher than those detected at room level. Furthermore, concentrations capable of causing pathology are much less than human sensory threshold levels.  Many design features affect room ventilation including the location, number, and configuration of supply and exhaust ducts.  Cage-level ventilation is further affected by the presence and/or type of filter top on the cage as well as the design and location of the cage relative to the room airflow pattern. Ventilation should be such that it keeps the concentration of waste gases to a minimum, reduces the spread of disease, provides a stable temperature and humidity and avoids drafts. Illumination Light intensity and photoperiodicity in animal rooms can affect reproductive function and animal vision.  The recommendation of the Guide for light intensity in animal rooms is 75- 125 footcandles (fc).  Variable light intensity control devices such as dimmer switches or multiple bank lighting can be installed to facilitate adequate light for observation and husbandry yet provide lower intensity light for general animal housing.  Cage position on a rack can be an important factor.  An 80-fold difference in light intensity can exist between the upper and lower shelf locations. Photoperiods or light/dark cycles (usually given in hours as L:D) can modify  reproductive behavior and circadian rhythms. A daily light cycle which has 12 to 14 hours of light is usually recommended for most species.  It is important to keep the light intensity and periodicity constant.  Animal rooms should be equipped with automatic light timers.  The presence of windows, either to the outside or to the corridor, can affect reproduction in some animals.  Corridor windows may be desirable for observational purposes, but they can provide enough light to affect circadian rhythms in nocturnal animals.  As with all environmental factors, the special characteristics of the animal should be taken into consideration when planning light cycles. Duration and type of light can affect estrus behavior.  Animals can have their reproductive cycles manipulated by changing the light cycle. This technique has been used in several rodent species, cats, and farm animals.  Reversed light cycles can be used to accommodate circadian rhythm, sleep and breeding studies within the normal working hours in an institution. Individual room timers provide a facility with more flexibility to meet a variety of experimental requirements. Noise (top of page) Excessive noise can also disrupt animal breeding behavior.  Noise at excessive levels can cause mechanical damage to the auditory system in both animals and man.  Some effects of noise in animals include audiogenic seizures, eosinophilia, increased serum cholesterol levels and increased adrenal weights.  Noise levels in animal facilities should not exceed 85 decibels (db). Food Standardized commercial diets are available for most laboratory species.  The Principle Investigator or Instructor is responsible for providing appropriate diets and ensuring that food is fresh and free from contaminants.  The IACUC can assist with selection of specialized diets and provide information on their availability. Bedding Bedding, such as hardwood chips, corn cobs and paper, are acceptable.  Pine shavings may be used as bedding material.  However, it has been documented that aromatic hydrocarbons from pine shavings or cedar bedding can induce hepatic microsomal enzymes.  Accordingly, such bedding may be inappropriate for animals involved in certain experiments. Handling The frequency and type of handling an animal receives is another non-experimental variable.  Investigators should be familiar with and skilled in the correct techniques for handling and restraining the species involved.  This can prevent injury to both the animal and the handler.  Daily husbandry routines may need to be scheduled around research needs. Close communication between the investigator and other personnel  can minimize handling stress.  For example, collection of biological samples may be performed during routine cage changing.  Because many animals are creatures of habit, regular handling may reduce stress. Sanitation (top of page) The Animal Welfare Act and the Guide have established schedules for frequency of cleaning animal rooms and for changing cages.  In some cases frequent cage cleaning may be disruptive to research objectives, as in the case of reproductive studies where frequent changes may eliminate pheromones necessary for reproduction.  Adding a small portion of bedding from the soiled cage to the fresh cage may prevent such problems while maintaining sanitation. Schedules can be altered to accommodate special research needs with prior approval of the IACUC. Waste Disposal Carcasses and animal wastes must be disposed of according to procedures approved by the IACUC.  Freezers are for deposit of animal carcasses which may then be periodically removed.  No food, supplies or materials other than animal carcasses or tissues should be placed in these freezers. Vermin Control The presence of pests in animal colonies can result in contamination of food and bedding, and the introduction of disease. The Principle Investigator or Instructor is responsible for maintaining a pest control program.  Various traps are recommended for controlling vermin.  Pesticides are used in animal areas only when necessary, and only after consultation with the investigator(s) whose animals will be exposed to them. Veterinary Care All animals should be routinely monitored for signs of pain or distress.  It is the responibility of the PI/CI to be familar with these signs to implement the appropriate protocol for treatment or eithanasia.  If veterinary care is required, it the responsibility of the PI/CI to make the necessary arrangements. Animal Procurement Newly acquired animals can introduce disease into established colonies.  To minimize the possibility of introducing disease into animal facilities, all arrangements for acquiring and housing live animals must be made with the explicit approval of the IACUC.  Animals may not be purchased or otherwise acquired until a fully approved animal use protocol is on file.  If wild animals are to be used, arrangements for any necessary quarantine must be made before animals are acquired.  The investigator is responsible for determining if permits (such as from the U.S. Fish and Wildlife Service or the Michigan Department of Natural Resources) are required.  Any necessary permits must be obtained before animals are acquired. Quarantine and Stabilization With some animals, a quarantine period may be necessary to minimize the introduction of disease into established colonies.  The extent of the quarantine period is determined by knowledge of the animals’ source and previous history. Arriving animals, regardless of source, should be allowed a stabilization period before use.  Such a period allows the animal to recover from shipping stress, adapt to the new surroundings, and become physiologically stable. Terminal procedures do not usually require a stabilization period. Separation of Species Physical separation of animals by species is generally recommended to reduce the possibility of transmission of latent diseases. This separation is usually accomplished by housing different species in separate rooms. Due to the limited nature of the animal care program (and facilities) at LSSU, concurrent projects will be limited to use of the same species. When animals of the same species are obtained from multiple sources, their microbiological status may differ, in which case separate housing may be advisable. Surveillance, Diagnosis, Treatment, and Control of Disease Principle Investigators and Instructors are responsible for checking all animals daily, including weekends and holidays, for clinical signs of illness, injury or abnormal behavior (Table 1).  In cases where such observation will interfere with experimental objectives, prior arrangements must be made to ensure adequate monitoring of animals and environmental systems. Emergency Care (top of page) Any health problem noted by any animal user at any time, including evenings, weekends and holidays, should be reported immediately to the IACUC.  This procedure should also be used to inform the IACUC of facilities malfunctions (e.g. excessively hot or cold animal rooms) which appear to directly threaten animal health. Anesthesia and Analgesia Animal procedures are reviewed by the IACUC to ensure that proposed anesthetics and analgesics are appropriate for the species and research objectives. The PI/CI should consult with a veterinarian to provide assistance with, or training in, the proper administration and use of anesthetics.  Other drugs may be approved in consultation with a veterinarian.  Muscle relaxants and paralytics are not anesthetics and cannot be used alone for surgical restraint.  They can be used in conjunction with other drugs which produce adequate anesthesia.  LSSU policy requires written documentation of all survival surgical procedures on animals, and the types and amounts of anesthetic, analgesic or tranquilizing drugs used.  This documentation should be maintained in or near the animal procedure area, and is subject to inspection by veterinary inspectors and the IACUC during its inspections of animal facilities and animal study areas.  The Guide requires that any proposal to conduct painful procedures without anesthesia or analgesia must be supervised directly by the PI/CI. Surgery Survival surgery is defined as any surgery from which the animal recovers consciousness.  Major surgery is defined as any surgical intervention that penetrates a body cavity or has the potential for producing a permanent handicap in an animal that is expected to recover.  Minor surgery is any operative procedure in which only skin or mucous membrane is incised (e.g. vascular cut down for catheter placement or implantation of pumps into subcutaneous tissues).  Also included are procedures involving biopsies or placement of probes or catheters requiring entry into a body cavity through a needle or trocar in combination with a minor surgical procedure.  Because they are minimally invasive, gonadectomies on rodents and lower vertebrates are usually considered minor surgical procedures. Multiple major survival surgery is defined as two or more major survival surgical procedures on a single animal.  It is permitted by the IACUC only under special circumstances, such as when the surgeries are essential and related components of a single study.  Cost considerations are not an adequate reason for performing multiple survival surgeries on an animal.  Appropriate aseptic techniques for these procedures include a clean uncluttered work area, preparation of the surgical site including clipping of the hair, disinfection of the skin and draping of the surgical site with sterile drapes, the use of sterile supplies and instruments and the use of sterile gloves and a surgical mask by the surgeon and any assistants working in the surgical field. Animals should generally be fasted overnight prior to anesthesia and surgery to prevent vomiting, aspiration, and problems associated with a distended intestinal tract. Animals must be observed to ensure uneventful recovery from anesthesia and surgery.  The animal must be monitored until it regains sternal recumbency and is capable of holding its head up, and should be returned to its regular cage only when it is alert, mobile, and breathing normally.  The animal should be kept warm and dry and fluids, analgesics and antibiotics administered as required.  Surgical wounds should be kept clean, and bandages or wound dressings changed as frequently as necessary to keep them clean and dry.  Subsequent care may include supportive fluids, analgesics, and other drugs as required; monitoring of the animal including clinical observation for signs of pain, abnormal behavior, appetite and excretory functions; providing adequate care of surgical incisions; and maintaining appropriate medical records.  CAUTION: Use of heat lamps and electric heating pads can result in severe burns or hyperthermia in animals that are anesthetized or otherwise unable to escape from the heat.  Close observation is required. Non-survival surgery is defined as any surgery in which the animal will not regain consciousness.  Such procedures may be performed in a suitably located and equipped laboratory, subject to IACUC evaluation and approval. Euthanasia (top of page) The Guide defines euthanasia as "...the procedure of killing animals rapidly and painlessly...".  LSSU euthanasia guidelines follow those established by the American Veterinary Medical Association Panel on Euthanasia. Proposed euthanasia techniques must be approved by the IACUC during review of animal use protocols.  Euthanasia should be carried out by personnel correctly trained in the method being used.  Measures should be taken to ensure that euthanasia is performed in a way that minimizes reactions among other animals that may be present.  Proper euthanasia includes a follow-up examination to confirm the absence of a heart-beat, which is a reliable indicator of death.  Monitoring respiration is not considered sufficient since the heart may continue to beat after visible respiration has ceased. Investigators should consult the AVMA classification of "Methods of Euthanasia by Species" from the "Report of the AVMA Panel on Euthanasia". Methods are summarized in: Table 2 - Euthanasia by Taxonomic Group Table 3 - AVMA Acceptable Method of Euthanasia Table 4 - AVMA Conditionally Acceptable Methods of Euthanasia Table 5 - AVMA Unacceptable Methods of Euthanasia