Appendix D.

Directions for the collecting and packing of material for despatch to
                    Muktesar for purposes of investigation

Blood, pus and other fluid, should be forwarded either in sealed
pipettes or as smear preparations on cover-glasses or slides. In using the
pipettes the following precautions are to be observed :—Both ends of
pipette to be sterilized in the flame (spirit lamp), one end to be
broken by means of a sterile forceps and inserted in the fluid, after which
the other end is to be broken. The fluid will then be drawn up into the
-pipette. Sometimes it is necessary to aspirate, but care should be taken that
fluid is not drawn into the mouth. After filling the pipette, seal both ends
in the flame. Two or three pipettes should be filled and after carefully
rolling in cotton-wool or other soft material, be packed in a box and for-
warded as early as possible.

To obtain blood from a living animal, a vein in the ear can be punc-
tured or an incision made into a muscle.

Care must be taken to wash the part and to sterilize with 5per cent
carbolic or other antiseptic fluid (not corrosive sublimate). The part
should be allowed to dry before making the incision, in order that the
blood may not be mixed with any of the antiseptic. If blood is taken from
an incision in muscle or skin, pressure should not be exerted on the part, as
in this way an excess of serum and few blood corpuscles are obtained.

In taking blood from a dead animal the most suitable method (except
in Anthrax) is to expose the heart, and after sterilizing an area by apply-
ing the heated blade of a knife, insert the pipette through this part into the
right ventricle of the heart.

In case of Anthrax, the blood is taken immediately after death from
vein of ear.

Smear preparation can be taken from any fluid, also from organs as
liver, spleen, etc. In such preparation, care must be taken that the material
is sterile and that the surface layer is as thin as possible. Thick smears
are useless. To take cover-glass preparations of blood or other fluid, a
drop, about the size of a pin head, is to be taken on one cover-glass and
immediately a second glass placed over this and allowed to rest for a
second, then drawn rapidly apart. No pressure should be exerted and no
delay made. If the blood corpuscles are drawn out, or fibrine deposited
on the glass, the preparations are useless. In the same method a prepara-
tion of fluid can be made on a slide by smearing the fluid by means of
the edge of a second slide or by means of a surgical needle. These prepara-
tions should be allowed to dry before packing. Each slide or cover-glass
should be carefully labelled. Before using care is to be taken that cover-
glasses and slides are clean.