40                              Appendix D.—(Contd.)

Organs, tissues and neoplasms should be taken as fresh as possible,
well washed in water and then placed in alcohol, rectified spirit, corrosive
sublimate (saturated solution) or formaldehyde (10 parts of the commercial
formaline to 90 parts water). Sections of more than one inch square are
not required for microscopical examination.

Cultures. — Culture tubes are inoculated by means of the platinum wire
or by use of pipettes. To take material from the interior of organs and
tissues, the surface is first sterilized by searing it with a hot knife which
has been heated in the flame of a spirit lamp and then a small incision is
made with another hot knife in the seared or sterilized area. The material
from the interior is collected on the end of the platinum wire, which has
also been previously heated in the flame. In the case of exudations care
should be taken not to contaminate such fluid by handling before the
material for culture has been obtained. The material is transferred by
means of the platinum wire to the surface of the culture medimum avoid-
ing the breaking of the surface. The material should be in small quantity
and be well distributed over the nutrient surface. A second tube should
be inoculated from the first, by touching the platinum wire previously
sterilized and cooled on the surface of the first tube, and rubbing the infect-
ed wire over the surface of the second tube.

At post mortems, cultures are made from heart, liver, spleen, lungs or
kidneys.

Culture tubes should be labelled after inoculation. Culture from the
blood of the heart should be made before the removal of that organ from
the body by scaring the right ventricle and then puncturing it with a
sterilized knife to admit the platinum wire in the pipette. The amount of
the blood used for the culture should be as much as will adhere to the
platinum wire, or one drop from a pipette.

In order to prevent contamination of the culture media, the rubber
cap should not be removed till tubes are required for use. After removing
the rubber caps, the mouth of the tube is sterilized in flame and after
inoculation of the media the cotton plug is slightly burnt in flame before
being inserted again in the tube. Before replacing the rubber cap after
inoculation of cultures, these should be sterilized by washing in aseptic
solution.

In case of Charbon Symptomatique a small piece of the infected
muscle should be dried and placed in a sterile test tube without any anti-
septic.