# **ANIMAL RESEARCH FACILITIES**

**STRATTON VETERANS ADMINISTRATION**

**MEDICAL CENTER**

**ALBANY, NEW YORK 12208**

**STANDARD OPERATING PROCEDURES**

**MARCH 2009**

### Table of Contents

1.0 INTRODUCTION 4

2.0 GENERAL STAFF POLICIES 4

2.1 Reporting Animal Concerns and Whistleblower's Policy 4 3.0 SECURITY
IN THE ANIMAL RESEARCH FACILITY 5

4.0 PER DIEMS AND SPACE 6

5.0 STAFFING 6 6.0 TRAINING OF STAFF 6

7.0 SAFETY AND THE USE OF PROTECTIVE CLOTHING 7

8.0 WASTE DISPOSAL 8

9.0 PEST CONTROL 8

10.0 ANIMAL PROCUREMENT 8

10.1 Animal Procurement procedures 9

10.2 Approved Vendors 9

11.0 ANIMAL RECEIVING PROCEDURES 9

11.1 Rodents 10

11.2 Rabbits 10

12.0 IDENTIFICATION OF ANIMALS HOUSED IN THE ANIMAL FACILITY 10

12.1 Identification Cards For Animals Being Bred 11

13.0 ANIMAL HOUSING 11

14.0 SOCIAL ENVIRONMENT AND ENRICHMENT 11

15.0 RABBIT CARE AND FEEDING 12

15.1 Schedule For Newly Acquired Rabbits 13 16.0 RODENT CARE AND FEEDING
13

17.0 GUINEA PIG CARE AND FEEDING 14

18.0 RECOMMENDED RELATIVE HUMIDITY, DRY -BULB TEMPERATURE for

COMMON LABORATORY ANIMALS 15

18.1 Temperature And Light Monitoring Of The Animal Rooms 15

18.2 Animal Room Environmental Logs 16 18.3 Room Activity Log 16

19.0 HANDLING SICK, DEAD, ESCAPED OR UNIDENTIFIED ANIMALS 16

19.1 Sick Animals 16

19.2 Dead Animals 16

19.3 Escaped or Unidentified Animals 17

20.0 ANIMAL ROOM SANITATION 17

20.1 Rabbit Rooms 17

20.2 Rat and Mouse Rooms 17

20.3 Guinea Pig Rooms 17

20.4Mopping Procedures 17

20.5 Room Breakdown 17

20.6 HVAC Filters 18

21.0 EQUIPMENT WASHlNG 18

21.1 Machine Operations 18

21.2 Cage & Rack Washing 18

22.0 FEED AND BEDDING HANDLING, STORAGE AND PREPARATION 19

23.0 SEMI-ANNUAL SEROLOGY TESTING ON RATS &MICE 20

24.0 EUTHANASIA 21

24.1 Approved Methods of Euthanasia for Mice, Rats and Guinea Pigs 21

24.2 Approved Methods of Euthanasia for Rabbits 22

25.0 SURVIVAL RODENT SURGERY 22

25.1 Preparations of the Surgeon 23

25.2 Gloving 24

26.0 ASEPTIC SURVIVAL SURGERY IN ANIMALS

OTHER THAN RODENTS 24

26.1 Patient Preparation 24

26.2 Cleaning & Disinfection of the Operating Room & Its Equipment 24

26.3 Operating Room Attire 25

26.4 Preparations of the Surgical Team 27

26.5 Technique 27

26.6 Toweling 27

26\. 7 Gowning 28

26.8 Gloving (Closed Method) 28

26.9 Post Operative Care 28

27.0 REFERENCES 29

ADDENDUM A -- Controlled Substance Procedure in the ARF 30

**1.0 INTRODUCTION**

The Stratton VA Medical Center Animal Research Facility (ARF) maintains
full accreditation with the Association for Assessment and Accreditation
of Laboratory Animal Care International (AAALAC). The Medical Center
actively supports the use of animals in research, teaching and testing.
However, the use of animals in VA research is a privilege granted with
the understanding and expectation that such research is conducted
according to the highest ethical and legal standards. These policies are
written to set forth the principles and procedures that govern research
activities involving laboratory animals in the Stratton VA Medical
Center Animal Research Facility.

**2.0 GENERAL STAFF POLICIES**

The general policies for all staff working within the Animal Research
Facility are as follows:

A\) The introduction of food and/or beverage into the ARF is strictly
prohibited except in

designated areas. Designated areas are offices and the break room only.
The

disposal of food and beverage refuse will only occur in these areas.

B\) Smoking is not allowed in any area of the ARF. This policy is
strictly enforced.

C\) Personal entertainment devices, such as stereos, radios and
televisions are

prohibited in all animal care areas. Special permission may be granted
by the

Associate Chief of Staff for Research and Development (ACOS/R&D) to use
these

devices in the operating room, necropsy room or procedural rooms. These
devices

are allowed in offices and the break room.

D\) To protect the health of the animals housed in the ARF, non-research
animals are

prohibited from all areas of the ARF. This prohibition includes pets of
any species.

E\) Any behavior by research personnel that threatens the health and
well-being of the

research animals is regarded as a serious infraction and will be
reported to the

proper authorities.

F\) All research personnel must have their identification cards
displayed while working

in the ARF.

G\) Any injuries or illnesses that occur to research personnel in the
ARF, as a result of

animal contact or exposure, must be reported to their Supervisor and the
Personnel

Health Unit.

H\) Unauthorized persons will not be allowed in the ARF without proper
ID and/or

permission of the ARF supervisor.

**2.1 Reporting Animal Concerns and Whistleblower Policy**

The Stratton VA Research Service is committed to the humane care and use
of laboratory animals. To ensure that laboratory animals receive humane
care and use/treatment in accordance with the highest ethical standards,
laws, regulations and polices governing animal research, the
Institutional Animal Care and Use Committee (IACUC) must review and if
warranted, address any animal-related concerns by the public or by the
Medical Center employees. The IACUC must review each concern in a timely
and systematic manner and when necessary take prompt and appropriate
corrective action.

Reports of animal welfare concerns may be made anonymously, if desired.
However, if the complainant would like to know the resolution of the
investigation, he or she must provide a name. All reports will be
handled confidentially; however, anonymity cannot be guaranteed.

Any concerns or deficiencies in the care and/or treatment of animals or
any activities related to animal care that may be improper or inhumane,
should be reported. Concerns may be reported to one's direct supervisor,
the Attending Veterinarian, the Associate Chief of Staff for Research
and Development, the IACUC Chair, or the ARF Supervisor. Reporting may
be done verbally, in writing or by e-mail. When reporting an incident,
factual information should be detailed in the report.

The Notification and Federal Employee Antidiscrimination and Retaliation
(No FEAR) Act protects individuals from retaliation for whistle blowing.
If an individual believes that they have been retaliated against for
whistle blowing, a formal complaint can be filed with the Human
Resources Department of the Medical Center. There are numerous signs
throughout the ARF detailing the Medical Center's No Fear Act/Whistle
Blowing Policy and the steps for reporting concerns.

**3.0 SECURITY IN THE ANIMAL RESEARCH FACILITY**

Vulnerability of the Animal Research Facility and staff is of high
priority and constant assessment is done to help assure a safe and
secure workplace for personnel and animals used in research. These
measures include:

A)  Background checks on new research personnel by Human Resources.

B)  Identification card swipe system on all external doors of the ARF.
    Whenever a

> person swipes into the facility, the Police Unit can instantly
> identify the card that
>
> is being used.

C)  Safety screens are located on all windows. In addition, these
    windows have

> alarms that detect movement of the window and loud noise levels in the
> area of
>
> the window. These alarms go directly to the Police Unit.

D)  There are five emergency buttons located in the hallways of the ARF.
    These buttons can be pushed for any emergency or potential
    threatening situation. The Police Unit will respond immediately to
    any incident in which a emergency button is pressed.

E)  All ARF's external doors are equipped with security hinges to
    prevent the door from being removed from the frame.

F)  A security camera is located on the main building directed at the
    two main entrances of the ARF. The Police Unit is responsible for
    monitoring the security camera.

G)  The Police Unit is responsible for verifying all external doors are
    secure during non-tour duty hours during each police shift.

**4.0 PER DIEMS AND SPACE**

All investigators utilizing animals in the ARF are charged a per diem
rate for the daily care of each individual animal. The per diem charge
includes feed, bedding, changing pans and cleaning cages, the
veterinarian technician's time to perform these duties and the
veterinarian costs. The rates are determined by the type of funding the
Investigators are receiving: VA Funded or Non-VA Funded research. The
current rates for each individual species per day are as follows:

VA FUNDED NON-VA FUNDED

Mice \$0.11 \$0.26

Rats \$0.28 \$0.56

Rabbits \$1.60 \$2.74

Guinea Pigs \$1.25 \$2.03

All Non VA Funded investigators are charged for space used at the Animal
Research Facility. This includes the rooms that are used to house their
animals, as well as laboratory and office space. Investigators are
charged a monthly fee of \$17.50 per square foot for all support areas.
Rooms that are used for the housing of animals are assessed for the
capacity of animals that requiring housing before the Investigator is
charged. The investigator is charged \$17.50 per square foot, pro-rated
according to the percentage of animal capacity in the housing areas.
These calculations are done on a daily basis and are charged to the
investigator monthly.

**5.0 STAFFING**

The Animal Research Facility has two animal technicians, the ARF
supervisor and a

part-time animal caretaker. They provide full coverage for the facility
during the weekdays, weekends and holidays. The ARF supervisor's regular
tour of duty is Monday-Friday. The regular tour of duty for the
caretaker includes weekends and holidays. The consulting veterinarian
makes visits to the facility every two weeks and is accessible via phone
or e-mail at all times. The names and home phone numbers of the ARF
supervisor, consulting veterinarian and local veterinarian for
emergencies are posted prominently in the facility. After hours
emergencies are handled through Police and Security Service,
notification to the Research Department will occur through the use of
the Research Emergency Cascade.

**6.0 TRAINING OF STAFF**

The ARF Supervisor is responsible for training of all animal care
personnel working in the Animal Research Facility. This will be done
with the assistance and guidance of the attending veterinarian. Training
will consist of direct hands on training with the ARF supervisor or
veterinarian, the reading and understanding of research policies and ARF
SOPs as well as required web based training on the specific species
housed in the ARF. Continuing education for animal care personnel will
consist of yearly required web based training, research staff training
and attendance at local lab animal meetings or symposiums.

The ARF Supervisor and attending veterinarian must also complete yearly
web based training and attend research staff training sessions. The ARF
supervisor and attending veterinarian will be expected to attend some
lab animal meetings or symposiums as part of their continuing education.
The attending veterinarian provides training twice a year at research
staff meetings.

**7.0 SAFETY AND THE USE OF PROTECTIVE CLOTHING**

Standard Precautions are used by animal care personnel working in the
ARF and annual training is provided in compliance to these standards.

Material Safety Data Sheets (MSDS) sheets are located in room 125 for
all chemical agents used by animal care personnel. There are several
signs in the ARF announcing the location of the MSDS sheets.

There are 3 emergency eyewash stations and 3 emergency shower pull
stations located in the Animal Research Facility. Eyewash stations are
tested weekly by the ARF supervisor and recorded on the card attached to
the eyewash station. Shower stations are tested twice a year by
Engineering Service personnel and recorded on the card attached to the
shower stations.

There are 12 fire extinguishers and 4 fire alarm pull stations in the
ARF. All fire extinguishers and fire pull stations are clearly marked.
All fire extinguishers are checked monthly by the Safety Office. In case
of an emergency, all exit doors are clearly marked.

All ARF research staff must follow the VA's policy on the proper use and
disposal of needles and syringes. Needles are not to be recapped.
Syringes with attached uncapped needles will be dropped into puncture
proof containers for disposal. When containers are full, they are placed
in the hazardous waste area for pick up by EMS personnel. Adequate
animal restraint is used to reduce the chance of accidental needle
sticks. This may involve proper restraining devices and the use of
anesthetics.

The ARF has 5 stationary hoods that may be used for tasks involving
hazardous or dangerous materials. Hoods are certified once a year to
help ensure that they are functioning properly.

All animal care personnel working in the ARF must don clean scrub suits
daily, except under certain circumstances such as non-animal contact
assignments, in which case the technician may wear a full length,
buttoned lab coat over his or her clothing. Dedicated steel toed shoes
are worn by animal care personnel while performing tasks in the ARF.
Additional protective clothing is used by animal care personnel for
various assigned duties. These would include safety eyewear, gloves, ear
protection, respirators, aprons and gowns. Disposable gloves are used
whenever animal contact procedures are done and when handling animal
wastes.

All other research personnel working in the ARF, including visitors must
don full length, buttoned lab coats during their stay in the ARF.
Uniforms used in the ARF are NOT to be worn outside of the facility or
taken home. Laundry facilities are located in the ARF for washing and
drying of all lab coats and scrubs.

Signs are posted throughout the ARF to bring attention to the animal
species that are used in the facility. This is to prevent or reduce the
exposure of individuals who may be allergic to one or more species of
animals in the facility. The signs instruct them to report to the
Personnel Health Unit for further evaluation.

**8.0 WASTE DISPOSAL**

All soiled bedding and refuse are treated as hazardous waste and are
packed in biohazard boxes. The boxes are picked up by EMS personnel
every two weeks and shipped by a hazardous waste company to be
destroyed. Animal carcasses are red bagged and transported in accordance
with regulatory requirements. Animal carcasses are stored in a freezer
in room 137. EMS personnel pick up and box carcasses every two weeks and
a hazardous waste company will ship boxes to be destroyed.

Radioactive wastes are stored in radioactive barrels by the Radiation
Safety Officer and later shipped by the radiation safety officer.

**9.0 PEST CONTROL**

The object of the Animal Research Facility's pest program is to control
arthropods and rodent pests utilizing mechanical control techniques in
conjunction with a sanitation program and minimal application of
approved insecticides if applicable. All chemical pest control
substances must be approved by the attending veterinarian and the ARF
supervisor for animal safety.

The ARF supervisor is responsible for the facility's pest control
program. Rodent pest levels are monitored by examination of the facility
for rodent excrements and chewing on feed bags as well as examination of
sticky traps. Arthropod pests are monitored using roach traps. Traps are
placed in rooms and are counted monthly. The ARF supervisor keeps a
record of these numbers in the ARF office. If a severe case of vermin
infestation is noted in an animal room, the room will be emptied and
exterminated. Currently there are no insecticides used in the ARF.

**10.0 ANIMAL** **PROCUREMENT**

All animal purchases must be approved by the ARF supervisor and the
IACUC Chairperson. The reasons for this are:

A\) Animal care staff must have housing and care available when animals
arrive.

B\) Animal care staff must provide disease surveillance and vaccinations
on arrival.

C\) It ensures that someone is available to receive and provide proper
attention to newly

arrived animals at all times as required by provisions of the Animal
Welfare Act

D\) It ensures that there are an adequate number of animals available
for that particular

Animal Component of Research Protocol (ACORP) in a given study year.

**10.1 Animal Procurement Procedures**

Any laboratory animal used in the Animal Research Facility must be
acquired in accordance with Federal laws, regulations and policy. All
deliveries of live animals must be made directly to the Animal Research
Facility in room 126 unless special arrangements have been made and
approved by the Associate Chief of Staff for Research and Development.

The procedures for ordering animals are as follows:

A)  Prior to ordering animals, an email must be sent to the ARF
    supervisor and IACUC Chairperson for verification that there is
    space available in Animal Research Facility to house the animals
    being requested and to confirm that the number of animals requested
    are available for use, according to the approved ACORP.

B)  The email must contain the number of the approved ACORP.

C)  Once a positive response is received from the ARF supervisor and
    IACUC Chairperson, the purchase order may be placed. For VA and
    Albany Research Institute orders, the purchase order form must
    contain the title and number of the approved ACORP. Orders will not
    be placed until both authorizations have been obtained. This is
    necessary to ensure that there are no deviations from the protocol.

D)  If any animals arrive to the Animal Research Facility without prior
    approval, they will be refused and returned to the vendor at the
    expense of the Principal Investigator. All instances of
    non-compliance will be presented to and addressed by the IACUC.

E)  After the arrival of approved animals, the ARF supervisor will track
    the number of animals for the particular study to ensure that the
    study does not exceed the allotted animals according to the ACORP.

**10.2 Approved Venders**

All animal purchases will be made through approved vendors, unless
otherwise noted and approved by the IACUC Committee. The current list of
approved vendors are as follows:

RABBITS -- Millbrook Labs, Harlan and Covance.

RATS -- Harlan, Charles River and Taconic.

MICE -- Charles River, Taconic and Jackson Labs.

GUINEA PIGS -- Charles River and Elm Hill.

**11.0 ANIMAL RECEIVING PROCEDURES**

All deliveries of animals will be through the receiving Room #126. After
animal lab personnel have verified a shipment of animals, they will
notify Research Budget and Fiscal Assistant. After a shipment of animals
has been received, the room and equipment must be disinfected before
receiving additional animals. The technician is responsible for having
all cages of animals identified appropriately. Identification will
include species, strain, date received, weight or age, sex,
investigator, source and protocol number. The U.S.D.A. form will be
verified against the animal numbers for species under the Animal Welfare
Act and will be filed in the Supervisor\'s office. These forms will be
maintained for the period required by law. All incoming animals are to
have a quarantine period, unless animals received are pathogen-free and
from the same source. The length of the quarantine period will vary
depending upon the species, vendor source, and health status of the
animal and will be determined at the time of ordering.

**11.1 Rodents**

Box shipment of rodents will be placed on the table provided in Room
126. The animal lab employee receiving the animals will verify the
accuracy of the shipment before signing the invoice. The boxes
containing the rodents will be taken directly into an available
quarantine room and placed into the appropriate type of cages and
labeled with the following information: Investigator, date received,
birth date, strain, vendor, sex, weight when received and protocol
number. The employee will check rodents for nasal or ocular discharge,
as well as general overall health. The empty shipping boxes will then be
taken directly to the trash dumpster, located directly outside of the
Animal Research Facility. Rodents with an unknown health status will be
quarantined for a minimum of two weeks. Rodents from an approved vendor
source with a known health status will be placed in an existing colony
from the same vendor source. A stabilizing period of five days is
encouraged for those rodents from an approved source.

**11.2 Rabbits**

Rabbits are received in Room 126. Animal lab personnel receiving the
animals will verify the accuracy of the shipment before signing the
invoice. When rabbits are received they will be examined for ear mite
infections, nasal or ocular discharge, diarrhea and skin lesions. The
animal is then weighed. Animals are then taken directly to an available
quarantine room and are placed into the appropriate cage and labeled
with the following information: Investigator, date received, strain,
vendor, sex, weight when received and protocol number. Shipping boxes
are then taken to the trash dumpster, located directly outside of the
Animal Research Facility. Rabbits from the same vendor source may be
placed directly into the animal room or otherwise quarantined in an
available room. Rabbits with an unknown health status will be
quarantined for a minimum of two weeks.

**12.0 IDENTIFICATION OF ANIMALS HOUSED IN THE ANIMAL**

**FACILITY**

All animals are to be clearly identified upon receipt of shipment or at
the time of birth. Unique numbers are assigned to animals larger than
rodents.

Methods: All animals are identified by attaching a cage card to the cage
or box. The following information must be supplied on the cage card
where applicable:

1\) Protocol Number 5) Date Received

2\) Investigator\'s Name 6) Species

3\) Source 7) Weight or Age on Arrival

4\) Strain 8) Sex

It is the responsibility of the animal care staff to properly identify
the animals and make sure the identification is legible. It is also
their responsibility that the animals have the proper identification on
them at all times. Any missing identification cards should be reported
to the ARF supervisor as soon as possible.

**12.1 Identification Cards for Animals Being Bred**

Animals that are bred in the facility are identified by an attached cage
card to the cage or box. The following information is required on the
cage card where applicable.

1\) Identification 6) Source

2\) Date of birth 7) Investigator's Name

3\) Number of animals 8) Protocol Number

4\) Parents 9) Date of Receipt

5\) Sex 10) Species and Type

**13.0 ANIMAL HOUSING**

The primary enclosure of a cage or box provides the limits of an
animal's immediate environment, so this area must allow for the normal
physiologic and behavioral needs of the animal, such as urination,
defecation, maintenance of body heat, etc. The housing requirements for
species used in the ARF are as follows:

Mice -- Polycarbonate boxes, at least 7.5"x 11"x 5" with a maximum of
five animals per

box.

Rats -- Polycarbonate boxes, 10.5"x19'x 8' with a maximum of 3 animals
per box.

Rabbits -- Stainless steel or plastic cages with 1 animal per cage.
There must be at least

3 square feet per animal for rabbits weighing less than 4 Kg, at least 5
square

feet per animal weighing over 4 Kg.

Guinea Pigs -- Polycarbonate boxes, 10.5"x19"x 8", with one guinea pig
per box.

**14.0 SOCIAL ENVIRONMENT AND ENRICHMENT**

Animals housed in the Animal Research Facility require a certain degree
of social interaction and/or enrichment to reduce boredom as well as
encourage healthy and normal habits. When possible, social animals are
housed in groups to stimulate social interaction. In cases where animals
are housed alone, such as in rabbits or guinea pigs, enrichment toys are
used to encourage interaction. Mice, rats and guinea pigs are all given
Nylorbones. Mice are given toys that encourage nesting. Rabbits are
given Jingle Balls as well as Bunny Blocks which they interact with.

**15.0 RABBIT CARE AND FEEDING**

The ARF staff should clearly observe all rabbits in the rooms that they
are assigned. Particularly notice appetite, changes in attitude, amount
and makeup of feces, etc. Any changes should be brought to the attention
of the ARF supervisor, investigator and the veterinarian.

Rabbits are housed in individual cages. The size of the cage is mandated
by the Animal Welfare Act and must be adhered to (see 13.0 above).

If rabbit feed is kept in the animal room it is to be placed in
vermin-proof containers that have tight-fitting lids. Periodically such
containers are to be completely emptied and sanitized. When feed is
placed in these containers, the technician will write the date this is
done and milling date of the feed on a tag, located on the side of the
container.

Pans are changed twice weekly. At the time of changing, clean pans are
covered with about ¼ inch layer of hardwood Sani-Chip Rabbit cages are
changed and sanitized on a two week schedule.

Unless required by experimental protocol rabbits are fed ad libitum a
regular commercial rabbit feed. Feed hoppers are checked daily and
refilled as needed. Water is provided with attached water bottles. Water
must be checked daily to insure adequacy and cleanliness. Water bottles
may be \"topped-off" using the water source within the animal room.
Bottles must be returned to the same cage when \"topping\" them off.
However, water bottles are sanitized at least weekly. Diet is not fed
beyond 180 days after milling date.

When moving rabbits to a new cage, the ID card is first moved to the new
cage followed by the particular animal. This is done to avoid misplacing
or incorrect identifying animals. Caution must be used in handling
rabbits during changing of cages, so that none are injured. The
technician should observe the rabbit for general health, malocclusion,
overgrown nails and signs of ear mites. These problems will be taken
care of by the technician and brought to the attention of the
veterinarian. On a monthly basis, nails are checked or trimmed and
rabbits weighed and weights recorded on the back of the cage card by the
ARF staff. When rabbit cages are changed, the technician must be sure
that the proper cage card is placed on the clean cage.

When the technician has completed servicing the rabbit room, the floor
will be swept and hosed down daily. At least once a week the floor will
be mopped using a clean mop head. The mop water will contain two ounces
per gallon of a quatricide disinfectant added according to the label
directions. Mops, brooms, dustpans, and mop buckets are dedicated to
particular rooms. Trashcans in the rooms are lined with plastic bags and
will be emptied regularly.

Rooms are completely emptied and sanitized every three months using two
ounces per gallon of a quatricide, with the use of a pressure sprayer.

**15.1 Feeding Schedule for Newly Acquired Rabbits**

When newly acquired rabbits are received and placed in their individual
cages, they are not offered food until approximately 24 hours after
receipt. The amount of food is then increased over a five day period,
starting with about 25 grams on day one to about 125 grams by day five.
Water is available ad libitum.

First 24 hours \-\-\--No Food

Day 1 \-\-\-\-\-\-\-\-\-\-\--25 grams

Day 2 \-\-\-\-\-\-\-\-\-\-\--50 grams

Day 3 \-\-\-\-\-\-\-\-\-\--100 grams

Day 4 \-\-\-\-\-\-\-\-\-\--125 grams

Day 5 \-\-\-\-\-\-\-\-\-\--125 grams

**16.0 RODENT CARE AND FEEDING**

The animal technician should clearly observe all rodents in rooms for
which they are responsible for with particular attention to: appetite,
changes in attitude, coughing, hair coat etc. Any changes should be
brought to the attention of the ARF supervisor, the investigator and the
veterinarian.

Rodent caging at the Animal Research Facility consists of solid bottom
plastic cages with wire lids. Rodents are normally housed in groups,
unless the protocol deems otherwise. Rats are housed up to three per
cage and mice up to five per cage. The size of the cages are mandated by
the Animal Welfare Act, and must be adhered to. There are some
specialized metabolism cages which may be used on occasion.

If rodent feed is kept in the animal room it is to be placed in
vermin-proof containers that have tight-fitting lids. Periodically such
containers are to be completely emptied and sanitized. When feed is
placed in these containers, the date this is done as well as the milling
date is written on a tag, located on the side of the container A
standard rodent diet will be given ad libitum to all rodents unless a
different feed type or schedule is required by the investigator. Feed
hoppers will be checked daily and filled as necessary. Diet is not fed
beyond 180 days of milling date.

Water will be provided [ad 1ibitum]{.underline} unless another schedule
is required by the investigator. Water will be checked daily. Pint
bottles will be used for all rodent cages. When rodents are housed in
plastic cages care must be taken to insure that sipper tubes are long
enough, and no curved sipper tubes are used.

Water bottles may be topped off using the water source within the animal
room. Topped off bottles must be returned to the same cage. All water
bottles must be changed and sanitized at least once weekly for mice and
rats..

Plastic boxes will have approximately 1/2\" layer of Care-Fresh placed
in the bottom of the boxes. Boxes used for rats will be changed twice
weekly and for mice once per week Wire lids are sanitized weekly. Racks
holding rodent boxes are to be sanitized once every two weeks.
Ventilated racks are changed and sanitized on a monthly basis.

When changing cages, first remove the ID card from the dirty cage and
place it on the clean cage. Then remove the rodents and place them into
the clean cage. This is done to avoid misplacing or incorrectly
identifying the animals. When plastic cages are changed, the wire lids
may be transferred from one cage to the next, but at least once every
week the lids should be changed and sanitized. When rodents are
transferred from one cage to another, they should be observed closely
for any abnormality.

Every day after each room has been checked and serviced, the rooms will
be swept once a week or daily as a on needed basis. The floor will be
mopped at least weekly using a clean mop head. The mop water will
include two ounces per gallon of a quatricide disinfectant added
according to label instructions. Mops, brooms, dustpans, and mop buckets
are dedicated to individual rooms. Trash cans in rooms will be lined
with plastic bags and emptied regularly.

Rooms are completely emptied and sanitized every three months using two
ounces per gallon of a quatricide disinfectant with the use of a
pressure sprayer.

**17.0 GUINEA PIG CARE AND FEEDING**

The animal technician should clearly observe all the Guinea Pigs in the
rooms for that they are responsible for with particular attention to
changes in appetite, hair coat, activity and vocalization. Guinea Pigs
are housed individually in solid bottom cages with wire lids. The cage
size is mandated by the Animal Welfare Act, and must be adhered to (See
13.0 above).

Guinea Pig boxes are changed three times a week. Cages will have
approximately ½ inch of Care Fresh placed in the bottom. To change
Guinea Pigs cages, first remove the ID card from the old cage and place
it on the clean cage, followed by the movement of the Guinea Pig to the
new cage. This is done to avoid misplacing or incorrectly identifying
the animals. Wire lids are changed and sanitized weekly. Racks holding
cages are changed every other week.

Guinea Pigs are fed a standard guinea pig diet ad libitum in stainless
steel feeders. The feeders are changed weekly and sanitized. Each feeder
is completely filled at each feeding. If Guinea Pig feed is kept in the
animal room, it is to be placed in vermin-proof containers that have
tight fitting lids. Periodically such containers will be emptied and
sanitized. When feed is placed in containers, the date this is done as
well as the milling date is written on a tag located on the side of the
container. Diet is not fed beyond 90 days from milling date.

Water bottles are changed and sanitized daily. Topping off of Guinea Pig
bottles is not permitted.

Each day, after a room has been checked and serviced, the floor will be
swept. The floor will be mopped at least twice weekly, using a clean mop
head. The mop water will contain two ounces per gallon of a quatricide
disinfectant added according to label instructions. Mops, brooms,
dustpans and mop buckets are dedicated to individual rooms. Trash cans
are lined with plastic bags and are emptied regularly.

Rooms are completely emptied and sanitized every three months using two
ounces per gallon of a quatricide disinfectant with the use of a
pressure sprayer.

**18.0 RECOMMENDED RELATIVE HUMIDITY, DRY-BULB**

**TEMPERATURE for COMMON LABORATORY ANIMALS**

> Relative Dry-Bulb Temperature

Animal Humidity(%) °C °F

\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_

Mouse 30-70 18-26 64.4-78.8

Rat 30-70 18-26 64.4-78.8

Hamster 30-70 18-26 64.4-78.8

Guinea pig 30-70 18-26 64.4-78.8

Rabbit 30-70 16-21 60.8-72.0

**18.1 Temperature and Light Monitoring Of The Animal Rooms**

Monitoring of the animal room environment is an essential tool for
controlling the environment.

High, low, and present temperatures (Fahrenheit) of each animal room are
recorded daily. Min/Max thermometers are to be used for monitoring. The
temperatures over the previous 24 hours should be recorded during the
same time period each day. The time of the recording is noted. In
addition, Data Loggers are used in the facility to spot check
temperature, humidity and light cycles in specific rooms and generate
reports of this activity. The Data Log System is a computer based
software system that uses satellite sensors that are placed in animal
rooms for a determined period. These sensors take environmental readings
at pre-determined time intervals and store this information. Afterwards,
the sensor is connected to a special adaptor in the computer located in
the ARF Supervisor's Office. The environmental data is then
electronically processed in the computer and a report is generated of
the continuous temperature, humidity and light cycles in the room for
that period. Temperatures recorded outside the ranges listed above for
the various species must be reported to the Engineering Service. Monthly
reports must be on file in facility office and available for review upon
request.

A 12-hour light, 12-hour dark cycle is maintained unless otherwise
requested by the investigator.

**18.2 Animal Room Environmental Logs**

Current animal room environmental records are maintained in each animal
room. It is a check list for the technician assigned to the room to
complete daily. The log consists of columns for the technician to mark
off the critical elements of daily husbandry, such as feeding and
watering have been done. There is an additional area to record the high,
low, and present temperatures and humidity. Time, date and the
technician's initials are recorded on the log after the work is
completed. Completed animal room environmental records are maintained in
the Animal Research Facility office for at least three years.

3.  **Room Activity Log**

Current room activity logs are maintained in each animal room. It
consists of a check list for the technician assigned to each room to be
filled out daily. There are columns to check off for the cleaning of
sinks, feed barrels, floors and surfaces, the changing of animal pans,
boxes and cages and room sanitation. The technician will initial the
room activity log after completion of duties in the room. Completed room
activity records are maintained in the Animal Research Facility office
for at least three years.

**19.0 HANDLING SICK, DEAD, ESCAPED OR UNIDENTIFIED ANIMALS**

Animal Technicians are to observe their assigned animals for signs of
disease or abnormal behavior at the beginning of each workday. If any
animal is appears to be sick or dead, or has escaped or is unidentified
the following steps should be taken for each respective situation.

**19.1 Sick Animals**

A sick animal report sheet will be filled out with the date, time,
species, animal ID, room number, cage location, investigator and
symptoms. The ARF supervisor will be notified and will check the animal.
The investigator and the staff veterinarian will then be notified. The
name of the person notified will be recorded. Only in exceptional cases
will the Veterinarian institute treatment without investigator
knowledge. All observations, medications, and treatments given will be
recorded on the sick animal report sheet and will be reviewed by the
veterinarian.

**19.2 Dead Animals**

A dead animal report will be filled out with the date, time, species,
animal ID, room number, cage location, investigator and date arrived.
The ARF supervisor and the veterinarian will be notified, as well as the
investigator. Upon request of investigator, dead animals will be
necropsied by the veterinarian or the ARF Supervisor. All necropsy
findings, as well as possible cause of death, will be recorded on the
dead animal report and reviewed by the veterinarian.

Dead animals will be stored in the carcass freezer located in room 137
of the Animal Research Facility. EMS personnel will pick up and box the
carcasses every two weeks and a hazardous waste company will ship boxes
to be destroyed.

**19.3 Escaped or Unidentified Animals**

Animals do escape from their cages. If such an animal is found free or
unidentified in a room, it will be placed in a new cage and identified
as "FOUND." Each investigator with animals in that room will be notified
in writing. Unclaimed animals will be euthanized 7 days after they are
found. Escaped animals found by investigators or staff should not be
placed in any cage except an empty cage. If an animal is found loose,
the ARF supervisor must be notified.

**20.0 ANIMAL ROOM SANITATION**

**20.1 Rabbit Rooms**

Rooms are swept and hosed down daily during the week. The rooms are
mopped weekly with two ounces per gallon of quatricide using a freshly
laundered mop. Every three months, rabbit rooms are completely emptied
and all walls, floors, and fixtures are sanitized using two ounces of a
quatricide disinfectant with the use of a pressure sprayer.

**20.2 Rat and Mouse Rooms**

Rooms are swept daily as needed but at least once weekly. The rooms are
mopped weekly with two ounces per gallon of a quatricide, using a
freshly laundered mop. Every three months, rat & mouse rooms are
completely emptied and all walls, floors and fixtures are sanitized
using two ounces of a quatricide disinfectant, with the use of a
pressure sprayer.

**20.3 Guinea Pig Rooms**

Rooms are swept daily and mopped twice weekly with two ounces per gallon
of a quatricide disinfectant using a freshly laundered mop. Every three
months, all guinea pig rooms are completely emptied and all walls,
floors and fixtures are sanitized with two ounces per gallon of a
quatricide disinfectant with the use of a pressure sprayer.

**20.4 Mopping Procedures**

2 ounces per gallon of a quatricide is applied to the floors of the room
and left for ten minutes. The mop is rinsed of detergents and the floor
is wet-mopped with fresh water.

**20.5 Room Breakdown**

For rodent and rabbit housing rooms, once every three months, all
animals dedicated implements and feed barrels are removed. All surfaces
are cleaned with two ounces of a quatricide disinfectant with the use of
a pressure sprayer and then rinsed with water.

**20.6 HVAC Filters**

For rodent housing rooms, filters are changed at least once a month by
EMS personnel.

For rabbit housing rooms, filters are changed at least twice monthly by
EMS personnel.

**21.0 EQUIPMENT WASHING**

The operations in Room #132, the ARF wash room, cover the cleaning of
most non-surgical equipment including cages, pans, boxes, water bottles,
etc. The room is operated for dirty items while clean items are placed
in the clean cage storage area.

At the start of operations the fan hood in the rack washing area is
started. This ensures a proper airflow pattern. Dirty items are to be
brought into the room only through the double door at the corridor C-50
(South Corridor)

**21.1 Machine Operations**

Most non-sterile equipment is sanitized in a Northstar R620 Cage & Rack
washer, located in room 132. It is a floor-mounted, stainless steel,
single door unit with a 6 foot ramp for loading and unloading large
animal cages and racks. Its rotary spray design is also capable of
cleaning boxes, bottles and utensils used in the care and housing of
research animals. All operations are programmed by computer and the user
only needs to turn the machine on and select pre-set wash and rinse
cycles. There is also an acid cycle used for the removal of urine scale.
The detergent used for the rack washer is Alka Det2 and for scale
removal, Acid Power is used. Both products are manufactured by Pharmacal
Research.

The cage and rack washer has scheduled maintenance quarterly from an
outside company, LBR Scientific. Records of all work performed on the
washer are kept in the ARF Office.

Temperature strips are used to verify that the water temperature reaches
the required 180 degrees. They are attached to an item being washed
during each use of the machine. These strips are placed on monthly
calendars and are on file in the ARF Office.

Every two weeks, the rack washer screens are taken out and cleaned of
debris by the ARF staff. This is done by removing the slotted stainless
steel floor panels of the machine and then removing the four wire mesh
screens located underneath. These screens are hosed down in the washroom
of all debris and replaced in the rack washer.

**21.2 Cage & Rack Washing**

Dirty items to be sanitized are brought into the washroom corridor C-50
(South Corridor). Pans and boxes containing dirty bedding are dumped
into infectious waste boxes located under the fan hood, near the C-50
corridor. Cages and racks are then pushed into the washer via solid 6
foot ramp. Boxes, pans, bottles and utensils can also be loaded on
special racks and then placed in the washer. At this time, the
temperature strip is placed on an item in the washer. The technician,
after closing the stainless steel door, will select which wash and rinse
cycles that are needed to sanitize the items inside. The different
cycles used in various equipment cleaning are clearly displayed on the
control panel of the rack washer. The start button is then pressed for
the machine to start filling and the cycles to begin. When items in the
washer are finished, the technician will unload the machine and take the
clean material out of the north end washroom door into the clean cage
area of the facility. Equipment is dried and stored in this area. Boxes
and pans are filled with bedding there as well.

Technicians are supplied with safety equipment such as gloves, eyewear,
steel toed shoes, ear protection and aprons for use while working in the
washroom.

Boxes containing soiled bedding are picked up by EMS personnel and then
transported by a waste removal company for disposal. New boxes for
soiled bedding are made on site as needed.

At the end of each day, the following shall be accomplished:

1\. The power switch is turned off on the Cage and Rack washer.

2\. The floor will be wet down, detergent put on floor and the floor
brushed down. The

floor is then hosed down completely.

3\. All floor drains will be emptied of debris daily.

4\. No dirty equipment should be left overnight unless absolutely
necessary. This is to

prevent vermin infestation. When equipment is left overnight it may be
stored in the

\"dirty\" end of the washroom and the washroom doors are closed

5\. The exhaust fan will be shut off at the end of the day, unless there
is dirty equipment in

the washroom.

6\. Microbial evaluation will be done monthly by randomly sampling
equipment taken out

of the rack washer.. A sterile culture swab is used for sampling and
then brought to the

VA Microbiology Lab for testing.

**22.0 FEED AND BEDDING HANDLING, STORAGE AND PREPARATION**

Standard animal diets and bedding will be purchased from known suppliers
using regular VA purchase procedures. The types of diets and bedding
used will be recommended by the ARF supervisor and approved by the
consulting veterinarian. Orders will be placed with Supply Service early
enough that feed and bedding are always available.

Our standard diets and bedding are currently being purchased from
Scott's Distributing Inc., Hudson, NH. Rabbit, rodent and guinea pig
feeds are Purina Pro-Lab diets. Contact bedding is Presspak Care Fresh.
Non-contact bedding is Hardwood Sani-Chips.

Special diets and ingredients for diets will be ordered by the
responsible investigator after discussion with the ARF supervisor and
consulting veterinarian. ARF personnel should keep the investigator
informed concerning the amounts on hand and the need to reorder.
Particular attention should be paid to feeds or ingredients ordered
through Albany Medical College or Albany College of Pharmacy to allow
adequate time for delivery.

All feed, bedding and ingredients will be received through the receiving
foyer, Room #126. Items will be checked against the Purchase Order for
completeness. Damage will be noted and feed and/or bedding delivered in
damaged containers will not be accepted. Signs of contamination or
spoilage will be checked and dated items will be verified to assure
currency.

Bulk animal feed and bedding will be placed in Room #141, stored in the
original sacks, bags, etc. Feed will be rotated to assure freshness.
Feed not date coded will have the date of receipt written on the bag.

Ingredients for diets which need to be refrigerated will be placed in
the refrigerator in Room #139, or other refrigerator space as available.
Care must be taken that such ingredients are not stored with materials
that could cause contamination of the ingredients.

Feed may be transferred into bins or cans when available and these may
be kept in the animal room. The containers should always be kept tightly
closed to prevent vermin infestation and periodically the container
should be emptied and cleaned. These containers will not be transferred
from room to room, and when the original bag is not in the container,
the type of feed will be identified on the top of the container lid. The
milling date and date the feed is placed in the container will be
plainly marked on the tag located on the side of the container

If ARF personnel are to prepare diet for investigators, it will be the
responsibility of the investigator to request such service in advance,
and instruct ARF personnel in the proper preparation of the diet.

**23.0 SEMI-ANNUAL SEROLOGY TESTING ON RATS & MICE**

Health monitoring and serology testing are done on mice and rats housed
in the ARF twice a year by the ARF supervisor. Virus-antibody free (V
AF/Plus) animals are placed in the same room as the appropriate research
animals for at least six weeks and then are anesthetized, bled (via
cardiac puncture) and sacrificed. The blood is then allowed to separate
and the serum is sent to Charles River Laboratories for determination of
the presence of any viruses, mycoplasmas or bacteria that have been
contracted from research animals. At that time, fecal flotations and
skin samples will also be taken from the sentinel animals to check for
internal and external parasites by the attending veterinarian or ARF
supervisor. After the results are forwarded to the Animal Research
Facility, the ARF supervisor will prepare a report for the attending
veterinarian, who will evaluate the report and take whatever steps
necessary to ensure the proper health status of animals in the facility.

During the six-week period, the sentinel animals will have contact with
the dirty bedding or where possible be housed directly in the same cages
as the research animals.

The sentinel animals will be anesthetized with Ketamine
(9Omgikg)/Xylazine (10mgikg) mixture until they reach a surgical plane
of anesthesia. After blood is collected via cardiac puncture, euthanasia
is achieved by an overdose of Ketamine/Xylazine. The blood is allowed to
coagulate at room temperature and is then centrifuged at 2000-3000 RPMs
for 15 minutes. The samples are then measured and diluted with
Phosphorate Buffered Saline (PBS) by mixing 1 part of serum to 4 parts
PBS. The samples are then packed in dry ice and shipped via overnight
service to Charles River Labs.

The number of animals used for monitoring purposes shall be no less than
12 mice and 10 rats per year and will be broken up into 2 groups, six
months apart. The numbers will depend on the number of animals being
housed and the number of rooms currently in use. A range of 1 sentinel
animal per 8 to 10 research animals will be used. Upon the
recommendation of the attending veterinarian, these numbers may be
altered based on the serology reports, visible signs of infection or
other health issues involving the Animals Research Facility.

The type and strain of sentinel animals will be as follows:

Mice: Swiss Webster, 13-15 grams, male and female, age range of 6 months
plus.

The vendor will be Charles River Labs.

Rats: Sprague Dawley (CD) 175-200 grams, male and female, age range of 6
months plus. The vendor will be Charles River Labs.

This procedure is required in order to maintain a high quality animal
care program and healthy animals.

**24.0 EUTHANASIA**

Euthanasia of animals used in the Animal Research Facility must be
performed in a manner that minimizes stress and discomfort to the
animals as well as reducing the undue distress to the persons performing
this task. All methods of euthanasia follow the recommendations of the
AVMA panel on euthanasia and any exception to these recommendations must
be project-specific, based on scientific necessity and require advanced
approval from the IACUC committee. The veterinarian, ARF supervisor and
trained animal care staff can carry out euthanasia procedures.
Investigators and their staff will have prior training and/or experience
verified by the IACUC upon review of the ACORP. If not, they must
consult the veterinarian prior to the procedure.

**24.1 Approved Methods of Euthanasia for Mice, Rats and Guinea Pigs**

A\) Carbon dioxide inhalation.

B\) Cervical dislocation under anesthesia for mice and small rats.

C\) Decapitation of anesthetized animal.

D\) General anesthesia followed by causation of death without regaining
consciousness

with the administration of barbiturate euthanasia solution.

E\) Intraperitoneal administration of pentobarbital at 3x the anesthetic
dose.

**24.2 Approved Methods of Euthanasia for Rabbits**

A\) Carbon dioxide inhalation with prior sedation.

B\) General anesthesia followed by causation of death without regaining
consciousness

with the administration of barbiturate euthanasia solution.

The use of carbon dioxide, compressed CO2 in cylinders, will be the only
source of CO2 used for euthanasia purposes. Death must be verified by
the person administrating the euthanasia prior to disposal of the
animal.

**25.0 SURVIVAL RODENT SURGERY**

Survival surgery on rodents does not require a special facility, but
must be performed using sterile instruments, surgical gloves, and
aseptic procedures to prevent clinical infections.

Instruments are to be sterilized by use of a cold sterilant (solutions
containing gluteraldhyde) or by autoclaving with steam or ethylene
oxide.

Surgical procedures will be performed in a designated area. The area
should not be cluttered to allow for easy cleaning. The area should be
cleaned prior to the procedure, as well as after completion of the
surgical procedure with a quaternary ammonia solution or other
equivalent solutions by the investigators or their technical staff.

The operative site on the animal should be prepared for surgery in order
to remove as many bacteria as possible without harming the skin or
interfering with wound healing. Skin cannot be sterilized; however,
bacteria can be reduced to a relatively safe level for surgery by
removal of hair, mechanical scrubbing, and the effective use of surgical
scrub and germicidal solutions. Hair removal may not always be necessary
dependant upon species or surgical site.

Hair should be clipped well beyond the margins of the incision. If the
animal is cooperative, the hair can be removed prior to anesthesia. A
number 40 clipper blade will remove most hair satisfactorily. If
additional hair needs to be removed, moisten the area with a soap
solution and shave with a straight razor. Hair can be removed close to
the skin when clipped in a direction opposite to that in which it grows.
The clipper blade should be checked prior to use for broken teeth, as
these cut and abrade the skin. The blade should be held flat and not
used in a raking motion, as this can injure the skin. Occasionally, the
blade will become hot, and it should be checked frequently to prevent
clipper burns. When the hair has been removed, the animal should be
thoroughly cleaned of loose hair.

A selection of compatible scrub and germicidal solutions is important to
achieve maximum effectiveness. The limitations of the solutions need to
be recognized. Alcohol solutions are not effective against bacterial
spores. A commonly used surgical scrub and germicidal solution
combination is povidone-iodine and 70 percent ethyl alcohol.

Povidone-iodine scrub should be applied with friction, followed by a
thorough rinsing with warm water. A more thorough preparation should be
performed after the animal is positioned on the operating table. The
surgical preparation should begin at the incision line and in a circular
motion proceed outward toward the periphery.

Discard the sponge when reaching the periphery or when either hair or
rectum has been touched. Alternate surgical scrub and germicidal
solutions at least three times and until the sponge with germicidal
solution remains clean after prepping. The last application of germicide
should be allowed to dry on the operative site for maximum
effectiveness.

The animal should be draped by the surgeon after gloving using sterile
technique. Draping should be done prior to instrument arrangement.

**25.1 Preparations of the Surgeon**

The objective of any scrub is to remove as many bacteria as possible by
mechanical scrubbing and the use of chemical antiseptics without harming
the skin. Many techniques are used so that every surface from the nails
to the elbow is thoroughly scrubbed.. The time and number of brush
strokes may vary depending on previous scrubs and on the type of
solutions used. Scrubbing times of 5 to 15 minutes and 10 to 20 brush
strokes per surface have been advocated. The combination of scrubbing
time and thorough brush strokes is the key to an effective surgical
scrub. A guideline for the thoroughness is 10 minutes or 10 brush
strokes per surface. Each scrub will include two scrubs and two rinses.

Fingernails should be short and free of nail polish. Rings and jewelry
should be removed. The scrub begins with a general soaping and rinsing
with warm water. The nails should be cleaned with a disposable nail
pick. The hands should be held higher than the elbows to prevent
contamination from water running off the elbows and dripping onto the
hands.

Skin surfaces should be scrubbed in a definite pattern of strokes in
order to reach all surfaces. Start the scrub at the fingertips,
proceeding to the surfaces of the fingers, the palms, the top of the
hands, and then to the arm surfaces. After one scrub is complete, the
hands and arms should be thoroughly rinsed. Discard the brush and
acquire a new one for the second scrub. The process is then repeated.

Drying the hands and forearms is necessary to prevent moisture from
becoming a possible source of contamination. Gloves can be donned more
easily with dried surfaces. Toweling should start at the fingers, then
proceed to the hand, and with a twist motion, dry the wrist and forearm
up to the elbow. It is important that a clean portion of the towel be
used for each section (i.e., right hand and arm, left hand and arm).

**25.2 Gloving**

Before gloving, lubricants can be applied to the hands to allow gloves
to slide on more easily. Powders have been replaced by gloving cream in
an effort to minimize dust particles in the operating room.

Gloves that are commercially pre-powdered, making additional lubricants
unnecessary, may be used. The outside of the gloves should not be
touched during the gloving.

**26.0 ASEPTIC SURVIVAL SURGERY IN ANIMALS OTHER THAN**

## RODENTS

**26.1 Patient Preparation**

The surgical patient should be prepared for surgery by withholding food
for at least 12 hours prior to surgery. Water should be allowed until
preoperative medication is given. Prophylactic antibiotics may be
required in a patient that has a risk of infection.

**26.2 Cleaning & Disinfection of the Operating Room & Its Equipment**

Each morning prior to surgery, the operating room should be damp dusted

with a 2 percent a quatricide solution. Walls, table surfaces, kick
buckets, chairs, and surgery lights should regularly receive, depending
on surgery schedule, a thorough disinfection. The surgery lights should
not be overlooked. Their positioning directly over the surgical field
can be a prime source of contamination. Wheels of equipment and various
foot pedals (i.e., power equipment suction, electro-surgical unit)
should be cleaned and disinfected. Places such as door ledges, cabinet
tops, towel dispensers, view boxes, and the underside of equipment
should not be overlooked.

Equipment such as the heating pad, the ECG, and the suction unit should
be

checked and ready for use. Instrument packs and supplies should be
assembled. A wheeled cart can be used for transporting packs and
supplies if they are stored outside the operating room.

Soiled instruments and discarded materials should be promptly removed
from the operating room. Tables, kick buckets, stands and heating pads
should be cleaned and disinfected after each case. Suction tubing and
bottles should be replaced with clean units. The floors should be mopped
with a 2 percent a quatricide solution.

All furniture and equipment in the operating room should be moved at the
end of the day for cleaning and disinfecting. A mop and bucket should be
reserved for the operating room and marked \"surgery only.\" The mop
should be cleaned and disinfected daily by being laundered or soaked in
a clean disinfectant for 30 minutes. Dry mopping, sweeping, and dry
dusting should not be done in the operating room as they tend to
increase air currents and the spread of organisms. After surgery, the
scrub sinks should be thoroughly cleaned and disinfected to remove oily
deposits that are left by the scrubbing process. The soap dispensers and
foot pedals should also be included in the cleaning and disinfecting
procedure. If the scrub brushes are reusable, they should be cleaned and
re-sterilized.

Monthly: Sterile supplies should be dated at the time of sterilization,
and this dating should be inventoried each month. All cloth or paper
articles are double wrapped and considered sterile for one month.
Articles in heat-sealed plastic are considered sterile for one year.

**26.3 Operating Room**

The operating room is an area of utmost cleanliness and constant effort
is required in maintaining asepsis. As persons entering the operating
room are potential sources of contamination, policies regulating attire
must be maintained. Surgical caps and masks must be worn whether surgery
is being performed or not. Cleanliness and good personal hygiene should
be habits of everyone who enters the operating room.

Visitors should be given clean observation gowns and disposable shoe
covers to be worn over their street clothes and shoes. Outer articles of
clothing, without additional gowns, should not be permitted in the
operating room.

The clothing of operating room personnel should be simple in design and
comfortable. Scrub tops and pants or scrub dresses are acceptable;
however, tops should be tucked in and dresses should fit closely. Loose
clothing can accidentally touch sterile surfaces. Cotton clothing is
preferred over silk, nylon, or other synthetics because of the explosive
hazard created by static electricity. Light blue, jade green, or misty
green colors produce less glare than white. If a separate pair of shoes
for surgery is impractical, shoe covers should be worn. All shoes must
be cleaned of blood and debris daily.

All hair must be covered. The type of hat worn is determined by the
amount of hair that needs to be covered. Those with short hair may wear
the standard cap. Others with fuller and longer hair should wear the
bouffant hat. Beards and sideburns can be effectively covered by
surgeon\'s hood.

A properly applied mask should fit snugly around the nose and chin.
Loosely fitting or improperly worn masks are of little value in
preventing exhaled organisms from entering the operating room. The mask
should be comfortable and easy to breathe through. Coughing, sneezing,
and talking increase moisture in the mask, and moisture decreases the
filtration of organisms and allows their escape into the operating room.
A moist mask is ineffective and must be replaced by a dry one.

The operative site should be prepared for surgery in order to remove as
many bacteria as possible without harming the skin or interfering with
wound healing. Skin cannot be sterilized; however, bacteria can be
reduced to a relatively safe level for surgery by removal of hair,
mechanical scrubbing, and the effective use of surgical scrub and
germicidal solutions. The best solution and technique remain
controversial. Hair removal and skin preparation should take place in
the designated animal preparation area.

Hair should be clipped well beyond the margins of the incision. If the
animal is cooperative, the hair can be removed prior to anesthesia. A
number 40 clipper blade will remove most hair satisfactorily. If
additional hair needs to be removed, moisten the area with a soap
solution and shave with a straight razor. Hair can be removed close to
the skin when clipped in a direction opposite to that in which it grows.
The clipper blade should be checked prior to use for broken teeth, as
these cut and abrade the skin. The blade should be held flat and not
used in a raking motion, as this can injure the skin. Occasionally, the
blade will become hot, and it should be checked frequently to prevent
clipper burns. When the hair has been removed, the animal should be
thoroughly vacuumed.

A selection of compatible scrub and germicidal solutions is important to
achieve maximum effectiveness. The limitations of the solutions need to
be recognized. For example, hexachlorophene loses its effectiveness when
regular soap or alcohol is applied afterwards. A commonly used surgical
scrub and germicidal solution combination is povidone-iodine and 70
percent ethyl alcohol.

The initial cleansing of the operative site should take place outside
the operating room.

Povidone-iodine scrub should be applied with friction, followed by a
thorough rinsing with warm water. A more thorough preparation should be
performed after the animal is positioned on the operating table. The
preparation solutions should be applied with sterile gloves or sterile
sponge forceps. The surgical preparation should begin at the incision
line and in a circular motion proceeding outward toward the periphery.

Discard the sponge when reaching the periphery or when either hair or
rectum has been touched. Alternate surgical scrub and germicidal
solutions at least three times and until the sponge with germicidal
solution remains clean after prepping. The last application of germicide
should be allowed to dry on the operative site for maximum
effectiveness.

The patient is draped by the surgeon after scrubbing, gowning, and
gloving. The patient should be draped prior to other preparation
responsibilities (i.e. arranging instruments, accepting sterile
supplies, etc.) in order to prepare a sterile field in which to work.
This prevents contamination of the gown by brushing up against the
non-sterile table edge.

The fenestrated drape should be large enough to cover the table and the
opening should be slightly larger than the length of the incision. A
single drape is unfolded and, by looking through the opening, is placed
directly over the incision site. Regardless of the method used, drapes
should never be dragged to the incision site. In four comer draping,
four drapes are applied with a double thickness of about 10 inches
toward the line of incision, with two drapes parallel to the patient and
two perpendicular to the patient. The first and second drape should be
placed parallel to the patient with the first drape applied closest to
the person draping in. A towel clamp may be used to temporarily secure
the drape to the patient. The third drape should be placed perpendicular
and cranial to the patient and secured with towel clamps. The fourth
drape should be placed perpendicular and caudal to the patient with the
end of the drape connecting to the Mayo stand. Towel clamps should be
used to secure the fourth drape to the patient and the Mayo stand.

**26.4 Preparations of the Surgical Team**

The objective of any scrub is to remove as many bacteria as possible by
mechanical scrubbing and the use of chemical antiseptics without harming
the skin. Many techniques are used so that every surface from the nails
to the elbow is thoroughly scrubbed.

**26.5 Technique**

The technique for a complete surgical scrub remains controversial. The
time and number of brush strokes may vary depending on previous scrubs
and on the type of solutions used. Scrubbing times of 5 to 15 minutes
and 10 to 20 brush strokes per surface have been advocated. The
combination of scrubbing time and thorough brush strokes is the key to
an effective surgical scrub. A guideline for the thoroughness is 10
minutes or 10 brush strokes per surface. Each scrub, however, should
include two scrubs and rinses.

Fingernails should be short and free of nail polish. Rings and jewelry
should be removed. The scrub begins with a general soaping and rinsing
with warm water. The nails should be cleaned with a disposable nail
pick. The hands should be held higher than the elbows to prevent
contamination from water running off the elbows and dripping onto the
hands. The scrub suit top should be tucked in and the body bent slightly
at the waist to allow dripping to run off into the sink and not onto the
scrub suit.

Skin surfaces should be scrubbed in a definite pattern of strokes in
order to reach all surfaces. Start the scrub at the fingertips,
proceeding to the surfaces of the fingers, the palms, the top of the
hands, and then to the arm surfaces. After one scrub is complete, the
hands and arms should be thoroughly rinsed. Discard the brush and
acquire a new one for the second scrub. The process is then repeated.

**26.6 Toweling**

Drying the hands and forearms is necessary to prevent moisture from
penetrating the gown and becoming a possible source of contamination.
Gloves can be donned more easily with dried surfaces. The towel should
be grasped at the top and allowed to unfold. The body should be bent at
the waist to prevent the loose end of the sterile towel from brushing
against the scrub suit. Toweling should start at the fingers, then
proceed to the hand, and with a twist motion, dry the wrist and forearm
up to the elbow. The dried hand should grasp the free end of the towel
and proceed to dry the opposite hand and arm. It is important that a
clean portion of the towel be used for each section (i.e., right hand
and arm and left hand and arm).

**26.7 Gowning**

The sterile gown should be opened prior to scrubbing by the surgeon or
other operating team members. The gown should be fan folded with the
inside facing outward. The entire gown should be grasped by holding the
inside edge of the neckband and allowed to unfold. Care should be taken
to prevent contamination of the gown by brushing against the counter
edge or other non-sterile objects. The hands should now be slipped into
the armholes in a upward motion. Avoid standing too close to the counter
edge or near a wall, as the sleeve edges and gown front could easily be
contaminated. Once the gown has been slipped on, it should be tied at
the neck by an assistant.

**26.8 Gloving (Closed Method)**

Closed method gloving reduces the potential for contamination, because
the bare hands never touch the gloves or gown sleeve edge. Before
gloving, lubricants can be applied to the hands to allow gloves to slide
on more easily. Powders have been replaced by gloving cream in an effort
to minimize dust particles in the operating room. Gloves are
commercially pre-powdered to make additional lubricants unnecessary.

Bare hands do not touch the gloves or the gown. Therefore, when gowning
do not allow fingers to go beyond the sleeve edge but pick up the right
glove through the gown with left hand. Place right glove on the arm with
the fingers pointing toward the shoulder and the thumb down. Through the
gown, use fingers to spread the cuff of the glove around the entire
right gown sleeve edge. Pull cuff back over sleeve edge. Slide hand into
glove by grasping the gown and pulling back. Repeat for the left glove.

**26.9 Post Operative Care**

It is the responsibility of the investigator to assure appropriate
post-surgical care of animals. Special post surgical care arrangements
can be made in advance with the veterinary technical service staff. This
care includes such procedures as maintenance of adequate fluid balance,
administration of antibiotics, analgesics, or other drugs whenever
indicated, the recording of rectal temperature, respiration rate and
pulse rate, and care of the surgical incisions, emergency treatment, and
similar clinical procedures. The duration of the post-surgical care will
vary with the type of surgery performed and the condition of the animal,
but it must be provided whenever it is needed, day or night.
Post-operative care should include the following items:

a\. Pulse and respiration determined every 5 minutes until the animal is
extubated.

b\. Pulse and respiration every 10 minutes until animal is sternal.

c\. Body temperature every 30 minutes until animal is standing.

d\. Extubation once swallowing reflex has returned and when there is no
longer a need

for either ventilatory support or oxygen administration.

e\. To reduce hypostatic congestion animals should be turned every 30
minutes, unless

this manipulation is contraindicated by the surgical procedure.

f\. Post-operative medications:

1.) continuation of fluids and antibiotic therapy where indicated.

2.) animals will be given analgesics routinely until there are no signs
of any

discomfort or obvious pain.

g\. Records should contain the above information, and should indicate
food and water

intake, urination, defecation, and care of the surgical wound. Standard
post-operative

records forms provided by the ARF should be used. Records of
post-operative care

must be maintained and should be available for inspection if necessary.
Post-operative

records must become affixed to the animal\'s clinical record sheet and
returned to the

ARF upon termination of the animal.

h\. Clinical observations must be recorded daily until sutures are
removed or animal is

fully recovered from the operation. i. Post-operative record keeping is
done by the Principal Investigator and/or the

technician. The records of surgery and post-operative surgery are
located in the ARF

supervisor\'s office

**27.0 REFERENCES**

**27.1 VHA Handbook 1200.7 (May 27, 2005)**

**27.2 Guide for the Care and Use of Laboratory Animals (copyright
2002)**

**ADDENDUM A -- Controlled Substance Procedures in the ARF**

# 

# 

# **CONTROLLED SUBSTANCE PROCEDURES**

# **IN THE ANIMAL RESEARCH FACILITY (ARF)**

**1. RESPONSIBILITIES**

> Controlled Substances (Schedules I-V, as defined in 21 USC Sec. 812 of
> 22 January 2002) may be used for animal research. They must be stored
> in a double-locked cabinet in Room 121 of the Animal Research Facility
> (ARF). The supervisor of the ARF or designee will have access to the
> cabinet and dispense all controlled substances.
>
> a\. The Associate Chief of Staff for Research (ACOS/R) or designee is
> responsible for ensuring compliance with the controlled substance
> procedures for animal research.

b.  Each dispensed controlled substance has a corresponding green sheet
    (VA form 10-2638) that is issued by Pharmacy Service. It must be
    used to record usage of the drug. Once a day, at the end of his tour
    of duty, the ARF supervisor will inventory all controlled substances
    in the cabinet and check the drugs dispensed and the corresponding
    green sheets for accuracy.

c.  When the ARF supervisor is unavailable, the ACOS/R and
    Administrative Officer for Research (AO/R) will have access to the
    keys, will dispense drugs according to the procedure outlined below,
    and will do shift counts. An Alcoholics and Narcotics Inventory and
    Certification Record Sheet (VA Form 10-1043) is then signed
    certifying the accuracy of the contents in the cabinet.

> d\. The expiration dates of the drugs will be checked against either
> the stamped dates on the vial or expirations noted in the product
> inserts. The label of the reconstituted drug will show the user's
> initials, the date when the solution was reconstituted, and expiration
> date indicated by the package insert once reconstituted. The facility
> will not use any expired drugs for animal research. Disposal of all
> expired drugs will be initiated by the ACOS/R or designee and returned
> to Pharmacy Service.

e.  To ensure compliance with the VHA Handbook 1108.2, the Controlled
    Substance Coordinator (CSC), appointed by the facility Director,
    will conduct unannounced monthly inspections in the ARF.

**2. PROCUREMENT AND DELIVERY OF CONTROLLED SUBSTANCES**

All controlled substances must be ordered and received through the
Research Office.

Investigators must submit a VA purchase order to the Research Office for
processing. No

other sources may be utilized to procure or attain controlled substances
for research.

Specific procedures will be followed within the Research Office to
procure and receive

these drugs as indicated below:

a.  The designated MD will be notified by the Research Budget clerk upon
    receipt of a request for purchase of a controlled substance.

> b\. The designated MD will review and approve the request, enter an
> electronic green sheet order (or equivalent) for the item and submit
> it to the Pharmacy Service for processing. If the item is not on an
> electronic green sheet order, a written VA prescription form will be
> submitted to the Pharmacy Service.
>
> c\. The pharmacy will process the order and complete the Drug
> Enforcement Agency (DEA)
>
> form 222 (if applicable) utilizing the Research DEA Certificate.
>
> d\. After receipt of the controlled substance, the pharmacy will
> contact the research office for
>
> delivery of the substance(s) and corresponding green sheet to the
> ACOS/R or designee.
>
> e\. The designated MD will electronically sign for receipt of the
> controlled substance(s) after
>
> verifying the identity of the controlled substance, strength, and
> quantity, via the electronic
>
> green sheet VISTA program or equivalent.
>
> f\. The product and corresponding green sheet will be given to the ARF
> Supervisor for
>
> placement in the Animal Facility Narcotic Area of Use (NAOU) in room
> 121.

**3. DISPENSING OF CONTROLLED SUBSTANCES**

The ARF supervisor, or designee, will dispense all controlled substances
to investigators and approved designees. Investigators or their
designees will be responsible for the accuracy of the recorded usage of
such drugs on the corresponding green sheet.

a\. The investigator or approved designee will request the controlled
substance from the

ARF Supervisor. The ARF Supervisor either dispenses the controlled
substance or

allows the investigator or designee to take possession of the controlled
substance.

b\. ARF Supervisor will make a copy of the green sheet to keep in his
possession if the

investigator maintains possession of the original green sheet and
controlled substance.

c\. Investigator or approved designee will sign and date the copy of the
green sheet when

they take possession of the drug. The controlled substance should never
be left in an

unsecured location when not in use while in possession of the
investigator.

d\. The species, protocol number, date, time, dose and balance shall be
recorded on the

green sheet at the time of each use, and the investigator or designee
shall sign the

document.

e\. When the drug and corresponding green sheet are returned, the ARF
supervisor

will verify the amount of drug remaining and secure the original green
sheet and drug

in the locked cabinet in Room 121.

**4. RECONCILIATION OF GREEN SHEETS**

a\. Green sheets with zero balance (no controlled substance remaining):

> 1.) ACOS/R or designee will flag green sheet as "ready for pick-up by
>
> pharmacy" by computer entry

2.  ARF supervisor disposes of empty containers.

b\. Green sheets with a balance (controlled substance remaining) and
expired controlled

substances.

> 1.) The ACOS/R or designee will coordinate product pick-up and green
> sheet
>
> transfer with a pharmacy representative.

c\. Green sheets with discrepancies:

> 1.) ARF supervisor and investigator will attempt to resolve the
> discrepancy and
>
> notify the CSC.
>
> 2.) The investigator will submit corrective action plan to the ACOS/R
> and the
>
> CSC.
>
> 3.) Any discrepancies unresolved will be forwarded to the ACOS/R and
> CSC for
>
> an investigation.