Freunds

1. First immunization injection can be Freund's Complete Adjuvant (CFA); however, due to the frequency of severe inflammation with this preparation, an alternative adjuvant should be used whenever possible (e.g.. Ribi Adjuvant System or Titermax).

Rabbits may be sedated with acepromazine during immunizations if desired.  This precaution protects personnel from accidental needle sticks which could result in painful lessons, and in the case of CFA, positive tuberculin tests.

2. Subcutaneous injections of < 0.1 ml (Mice) or 0.5 ml (rabbits) should be given at multiple injection sites (<5 sites/rabbit or mouse).  To minimize secondary infection, injection sites should be prepared by shaving the area and swabbing with betadine.  Alternatively, intraperitoneal injections of <0.1 ml can be used in mice.

3. Intradermal injection sites are not recommended in mice.  However, intradermal injections are allowed in rabbits at volumes of 0.05-0.1 ml/site.  Intradermal injection is not recommended with CFA due to ulceration.

4. Injection sites should be reasonable distributed (mice) and >2 cm apart (rabbits) to prevent coalescence of lesions and subsequent development of draining abscesses, etc.

5. Adjuvant injection into the footpads of rabbits is not allowed.  substantial scientific justification is required for footpad injection in mice.  In this event, footpad injections must be limited to one hind foot and requires the addition of soft bedding to the cage.

6. The number of injections, routes and total immunizations should be thoroughly described.

7. Veterinary staff should be notified if evidence of pain or distress or injection site lesions are observed.