water and NaOH and bring to boil, stirring. 
Cool, refrigerate overnight and skim off any 
remaining fat. Filter the mixture through two 
layers of gauze and spread the meat particles 
out to partially dry. 
Add sufficient distilled water to the filtrate to 
restore 1 liter original volume and add: 
Trypticase or peptone 
30 gm. 
Yeast extract 
5 gm. 
Potassium phosphate 
5 gm. 
Glucose 
3 gm. 
Adjust pH to 7.8 with 1 N NaOH. 
Dispense meat particles in 15 x 125 mm. 
screw-cap tubes with a small scoop and add the 
enriched filtrate. Use about 1 part meat par- 
ticles plus 3 to 4 parts liquid (v/v) per tube. 
Add a few iron filings to each tube. Tubes 
should be more than half full (about 8 ml. 
fluid). Autoclave at 121° C. for 15 minutes. 
APPENDIX III 
Acceptable Containers for Use in Shipping Specimens 
1. Unfrozen specimens 
For shipping small numbers of specimens (1 to 
3 ) of whole blood or serum, any one of the usual 
double-mailing containers with a metal bottom 
and metal screw cap is satisfactory, providing 
the cap is tight. The tubes should be individually 
wrapped and identified, and the identifying list 
should be wrapped around the outside of the 
inner case before this is inserted in the outer case. 
Sufficient absorbent cotton should be included 
in the inner case to absorb any liquid resulting 
from breakage in transit. 
Shipment of large numbers of tubes in a card- 
board carton is possible if the tubes are indi- 
vidually wrapped and sufficient absorbent buffer 
material — cotton, shredded paper, excelsior, etc. 
— is included between the individual tubes and 
next to the walls of the carton to absorb shocks 
and leakage. The conventional cardboard car- 
tons wiih partitions designed to create a com- 
partment for each tube are most acceptable. 
2. Frozen specimens 
Small numbers of frozen sera or tissue blocks 
may be shipped in metal thermos-type con- 
tainers or in cartons with dry ice or bags of Sno- 
gel or other reversible refrigerant. If the thermos- 
type container is used, it should be pre-chilled 
with dry ice for several hours before the speci- 
mens are introduced. The specimens should be 
individually wrapped and identified and packed 
in such a way that they will not become loose as 
the dry ice evaporates. Enclose the thermos in 
an outer carton containing additional dry ice, if 
necessary, and adequate insulating material to 
hold the temperature for at least 24 hours longer 
than the package is expected to be in transit. 
Small numbers of frozen specimens may be trans- 
ported short distances by placing them in insu- 
lated paper bags — the so-called “Jiffy Bag” used 
for ice cream — with a small amount of dry ice. 
These bags should be enclosed within one or two 
other “Jiffy Bags” of larger size. Frozen bags 
of “Snogel” may be used in place of dry ice. 
Larger numbers of tubes should be individually 
identified and wrapped and packed in cardboard 
cartons with dry ice above and below and 
an abundance of insulating material around them. 
Wrap the carton with several layers of heavy 
brown wrapping paper and tie or seal it securely 
with tape. This carton should be enclosed in a 
larger one which should be wrapped and sealed 
before shipment. The package should be shipped 
by the most rapid means available. 
3. Refrigerated but not frozen specimens 
Animal heads should always be refrigerated but 
not frozen during transportation to the labora- 
tory. If the head is that of a small animal, it 
may be placed in a friction top can of proper 
size, the lid firmly pressed into position, and the 
can enclosed in a large size lard can or other con- 
tainer. Cracked ice is used to fill the outer can 
and, if available, a small amount of dry ice may 
be included as this will help to hold the tempera- 
ture down for a longer time without actually 
freezing the specimen. 
4. Glass slides 
Whenever available, slide boxes should be used 
with tissue paper stuffed between the slides. 
Otherwise, the slides should be individually 
wrapped in tissue and shipped in a screw-capped 
mailing container with suitable absorbent ma- 
terial to prevent breakage. Under no circum- 
stances, should slides be mailed in a letter or man- 
ila envelope. 
40 
