USA > Ohio > Hancock County > Findlay > Twentieth Century History of Findlay and Hancock County, Ohio, and Representative Citizens > Part 134
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3
9am
4
9am
5 9am
Mich 16
9am
1pm
5pm
17
9am
18
9am
19 9 am
Mich 21
9am
- INOC , 4
1pm
5pm
22
9am
73
9am
24
9am
Apr G
9am
-INOC ALL
...
[No. 85
JOHNS HOPKINS HOSPITAL BULLETIN.
264
Mch 2 9 am
Total WBC
5pm
LM --
PMN.
3 9am
¥ 9am
5 9am
Mch 16 9am
1pm
Polymorphonuclears
5pm
3 9am
Rabbits Nos. 15, 24, 92 and 104 showing absolute values of poly-
1pm
values of polymorphonuclear, large and small mononuclear leuco-
Rabbit No. 104 showing total leucocyte counts and absolute
5pm
7 9am
& 9am
9 98%
1pm
5pm
PMN
Rabbit 15-
92-1
24 -------
Small Mononucleaus
24 9am
Apr 6 9am
+INOC
1pm
CHART IX.
19 9am
19 9am
Mch 21 9 am
Mich 21 9am
Apr.
1pm
small mononuclear leucocytes.
Rabbits Nos. 15, 24, 92 and 104 showing absolute values al
CHART XI.
CHART XII.
morphonuclear leucocytes.
SM -
bbit No. 15:
'otal leucocytes. -
+
+
+ + 1
++ 1
1 1+
+ ++
+
bbit No. 24:
'otal leucocytes. +
'mn
mn
bbit No. 92:
'otal leucocytes.
mn
mn
obit No. 104:
otal leucocytes.
mn
mn
+++
+ + 1
+
++ 1
| | + +
+ + 1
1 1+ +
- denotes an increase over the preceding count.
- denotes a decrease from the preceding count.
) denotes no change from the preceding count.
For the purpose of better comparing the changes in the ymorphonuclear and small mononuclear counts, we have pared Charts X and XI in which their curves have been erimposed. Chart X shows a definite rise in the poly- rphonuclears of all rabbits three hours after the intra- mal inoculation of 1/100 cc. serum; this rise, however, participated in by rabbit No. 15, which had not been viously injected, as well as by the others which had been cted intravenously on March 21. Chart XI shows noth- remarkable except the sharp rise in the mononuclears of oit No. 15 on April 6 at 5 p. m., which we are unable to lain.
harts XII, XIII, XIV, XV, show the percentage values he differential counts on rabbits Nos. 15, 24, 92 and 104, ectively. The black areas at the bottom of the charts esent the percentage of polymorphonuclears; the shaded s, the large mononuclears; the unshaded areas at the top he charts, the small mononuclears; while the unshaded s between the small and large mononuclears represent the iophiles and mast cells combined. These charts show the relative numbers of large mononuclears, eosinophiles mast cells do not undergo great variations, and that the ionship between the polymorphonuclears and small mono- ears is, in general, a reciprocal one; that is an increase le percentage of polymorphonuclears is accompanied by a case in the percentage of small mononuclears and vice
is important to realize the different impression one gets a consideration of the differential count expressed in ntages and in absolute values. To illustrate, suppose a leucocyte count of 8000 cells, of which the polymorpho- ars are 60 per cent, small mononuclears 30 per cent, mononuclears, eosinophiles, and mast cells combined 10 ent. Now imagine the total count to increase to 10,000
ibbit No. 15 served as a control and did not receive intra- s inoculation.
cells. The percentage values now change to 68 per cent for the polymorphonuclears, 24 per cent small mononuclears, and 8 per cent mononuclears, eosinophiles and mast cells combined. How little idea one could get of the real change from a con- sideration of the percentages alone can be seen from this example ;
Pmn
60% =4800
W. B. C. 8000 Sm
30 = 2400
Lm, Eos and Mst. 10 = 800
68% =6800
W. B. C. 10000 Sm
Lm, Eos and Mst.
24 = 2400 8 = 800
In order to confirm the results of the preceding series we ran through a third series, consisting of rabbits Nos. 105, 106, 107 and 108. Of these No. 105 served as a control and received on sensitizing injection; the remaining three received intravenous injections of normal horse serum on June 16 at 10 a. m. as follows :
Rabbit No. 106 1.0 cc. serum. Rabbit No. 107 2.5 cc. serum. Rabbit No. 108 5.0 cc. serum.
The total and differential counts are given in the following tables :
Rabbit No. 105
Total
Pmn
Lm
Sm
Eos
Mst
1910
Leucocytes % Total
%
Total % Total
%
Total
% Total
May 23, 9 a. m.
9900
26.5 2623
3.5
347
60.5 5989
4.5
446
5.0
495
1 p. m.
12140
28.5
8460
4.0
485
65.0 7891
1.5
182
1.0
121
5 p. m.
12900
27.5
3547
3.5
451
63.5 8191
2.0
258
3.5
451
24, 9 a. m.
9780
32.0
8129
5,0
489
59.0 5770
1.5
146
2.5
244
25, 9 a. m.
9080
22.0
1997
6.0
545
66.0 5993
69.0
5584
2.0
160
2.0
160
1, 9 a. m.
7540
21.0
1583
5.0
377
71.5
5391
1.0
75
1.5 113
1 p. m.
10320
24.5
2528
11.5
1187
62.0
6898
..
..
1.5
170
2, 9 a. m.
10500
16.0
1680
2072
2.5
285
71.5
6785
0.5
3.5
330 265
June 16, 9 a. m.
7720
36.0
2779
6.0
463
884
61.0
4880
5161
0.5
40
5.0
397
19, 9 a. m.
5300
26.5
1404
6.0
318
66.5 3524
..
1.0
53
June 29, 9 a. m.
8660
21.5 1861
4.0
846
69.0 5975
1.0
87
4.5
890
(Inoc. 0.01 cc. serum
intradermally.)
1 p. m.
10250
27.C
2819
3.0
813
65.5 6838
1.0
104 8.5
365
5 p. m.
11260
88.5
4335
4.5
507
55.0
6193
2.0
225
July
1, 9 a. m.
8580
25.5
2188
5.0
498
65.5 5607
85
8.0 2.0
257
2, 9 a. m.
7440
33.5
2492
2.5
186
62.0
4613
..
149
Rabbit No. 106 May 23, 9 a. m.
27.5
1858
309
64.0 8148
1.0
49
1 p. m.
29.5
2106
9.0
648
67.0
4824
1.0
72
24, 9 a. m.
6100
11.0
671
4.5
274
81.0
4941
152
1.0
61
25, 9 a. m.
5580
29.5
1646
978
12.0
550
62.5 2912
23
4.0
186
June
1, 9 a. m.
6000
22.5
1863
14.0
848
56.0 3393
0.5
90
7.0 5.0
438
5 p. m.
6500
36.0
2861
2117
10.5
706
51.0 3427
3, 9 a. m.
4860
32.0 1995
5.
218
55.5 2420
4, 9 a. m.
4860
47.5 2071
6.0
262
38.0 1657
1.0
43
7.5 7.5
327 327
June 16, 9 a. m.
4040
28.5 1151
7.0
289
55.5 2242
0.5
20
8.5 343
(Inoc. 1.0 cc. serum intravenously.)
1 p. m.
8300
47.5 3942
4.5
373
45.0
3785
3.0
249
5 p. m.
6440
80.5 1964
7.0
59.5
3832
3.0 198
17, 9 a. m.
8800
33.5 1273
6.5
247
54.0 2052
0.5
19
5.5
209
18, 9 a. m.
4760
45.5 2166
10.5
500
89.0
1836
2361
. .
6.0
295
June 29, 9 a. m.
5580
47.5 2650
7.0
390
40.0
2232
1.5
93 4.0
223
(Inoc. 0.01 cc. serum intradermally.) 6460 2389
6.5
420
55.5
3585
1.0
65
5 p. m.
6250
33.0
2062
10.5
656
46.0
2875
1.0
63
8.5
576
July
1, 9 a. m.
7390
37.5
2767
8.5
627
47.0
3469
1.5
111
5.5
406
2, 9 a. m.
4720
40.0
1888
6.5
807
49.0
2313
0.5
23
4.0
189
Rabbit No. 107
May 23, 9 a. m.
4800
38.5
1771
18.0
598
44.0 2024
..
..
4.5
207
1 p. m.
7480
29.5
2206
12.5
935
52.0
8889
6.0
449
5 p. m.
9440
27.5
2596
10.5
991
56.5
5333
0.5
17 5.0
472
24, 9 a. m.
4440
24.5
1088
13.0
577
60.0
2664
..
..
2.5
111
25, 9 a. m.
5580
29.0
1618
14.0
781
55.0
3069
. .
..
2.0
112
26, 9 a. m.
5840
32.5
1898
11.0
642
50.0 2920
..
6.5
380
..
..
..
5.0
238
19, 9 a. m.
4920
39.0 1919
7.0
844
712
35.5
2407
1.0
68
9.5 594
30, 9 a. m.
6780
44.5 8017
10.5
6.5
557
51.0
4447
..
..
8.0
825
.
. .
:
7.0
470
5 p. m.
8000
33.0
2640
2104
1462
5.0
840
71.5
4862
: : 8 : ..
5.5
577
4, 9 a. m.
10600
20.0 2120
5.0
580
72.5
7685
4362
0.5
88
1.0
77
1 p. m.
6980
89.5
2757
5.5
2.5
8.0
200 288
477
59.5 5676
4.0
382
80, 9 a. m.
9540
81.5
8005
5.0
749
58.0
4141
2.0
1.43
5 p. m.
7200
23.0
1656
11.0
614
55.5 3097
28
3.5
195
26, 9 a. m.
4660
21.0
37.5
3270
10.5
689
45.5
2985
2, 9 a. m.
6720
31.5
2608
5.5
624
7.0
785
71.5
7507
58.5 58.0
3699
..
3.5
280
17, 9 a. m.
7940
26.5
18, 9 a. m.
6800
21.5
136
26, 9 a. m.
8020
21.5 1724
5.5
441
70.0
7988
..
2.0
206
5 p. m.
11340
+ +1
++ 1
| |+
++ 1
+1+
1 1+
++ 1
0+ |
+ 1 +
I+1
++ 1
+ +0
March 21
March 2 March 16 April 6 1 p. m. 5 p. m. 1p. m. 5 p. m. 1 p. m. 5p. m. 1 p. m. 5p. m
'mn
O
+
Imn
+
+
1+ 1
2.0
181
4.0
363
June
3, 9 a. m.
9420
22.0
2.5
2.0
139
..
..
-
1.0 ..
..
..
. 2.5 0.5 0.5
424
1 p. m.
8720
1 p. m.
37.0
Digitized by
4920 7140
7.5 10.5
451
48.0
65.0
..
..
2.0
23.0
THIRD SERIES.
+
1
Digitized by
10
20
30
40
50
60
70
80
90
8
Mich 2 9am
SMN
Mch 3 9am
Meh ¥ 9 am
Meh 5 9am
Mich 16 9am
1 pm
5 pm
Mch 17 9am
Mch 12 9am
Mich 19 9am
Mich 21 9am
+Inoc.
1pm
5pm
Mch 22 9am
Mch 23 9 am
Mich 24 9am
Apr 6 9am
+Inoc.
1pm
5pm
Apr 7 9am
Apr 8 9am
Apv 9 9am
100%
90
80
70
60
50
40
50
20
10
small mononuclear leucocytes for entire period of observation.
10
20
30
3
70
80
¥
100
Mch 2 9am
1pm
PMN
SMN
Mch 3 9am
Mch 4 9am
Mch 5 9am
Mch16 9am
1pm
5pm
Mch17 9am
Mch 18 9am
Mich 19 9am
Mich 21 9am
-Inoc.
1pm
5 pm
Mch 22 9am
Mch 23 9am
Mch 24 9am
Apr 6 9am
4-Inoc.
1pm
5pm
Apr 7 9am
Apr '8 9am
Apr 9 9am
100%
90
80
70
60
50
#0
30
20
10
10
20
30
40
50
60
70
80
90
100.
1000
2000
3000
4000
6000
7000
9000
Rabbit 105
..
5 pm
108
106
25 9am
'26 9am
June / 9am
5pm
2
9am,
3
4
9 am
104 107. 208, June 16
9 am
1 pm
5pm
5 pm
17 9am
18 9am
19 9am
June 29 9 am
V
1 pm
small mononuclear leucocytes for entire period of observation.
Rabbit No. 104 showing percentage values of polymorphonucku large mononuclear, eosinophiles and mast cells combined, and
Mch 2 9am
1pm
PMN
SMN
5pm
Mol 3 9am
107
24 9am
Mch 4 Jam
Mch 1 9am
1 pm
5pm
Mch 17 9am
Mch /3 9am
Mch 19 9am
Mch 21 9am
Inec.
JOHNS HOPKINS HOSPITAL BULLETIN.
[No.1 J
266
CHART XV.
Mcx 22 9am
Men 13 9am
MA-324 9am
Ppr 6 9am
Inde-
Ipm
mal horse serum intradermally in all rabbits.
serum intravenously as follows: Rabbit No. 106, 1 cc .; Rabbit Na for entire period of observation. June 16 injected normal horse
CHART XVI.
107, 2.5 cc .; Rabbit No. 108, 5 cc. June 29 injected 0.01 cc. Der-
Rabbits Nos. 105, 106, 107 and 108 showing total leucocyte cours
may 23 9 am
large mononuclear, eosinophiles and mast cells combined, and
Rabbit No. 24 showing percentage values of polymorphonuclear,
5pm
PMN
small mononuclear leucocytes for entire period of observation. large mononuclear, eosinophiles and mast cells combined, and Rabbit No. 92 showing percentage values of polymorphonuclear,
CHART XIV.
CHART XIII.
Total WBC-
.
PWN. -
- -
CHART XVII.
bbit No. 105 showing total leucocyte counts and absolute s of polymorphonuclear, large and small mononuclear leuco- for entire period of observation.
.
Total wec
PMON
SM ..
-
CHART XIX.
Rabbit No. 107 showing total leucocyte counts and absolute values of polymorphonuclear, large and small mononuclear leuco- cytes for entire period of observation.
mai
MP6
"
2
June 16
June 29
July
Total war
PMM
SM
.. ...
LA
CHART XVIII.
it No. 106 showing total leucocyte counts and absolute of polymorphonuclear, large and small mononuclear leuco- or entire period of observation.
mas
, 9am 2 9am
Jane
4x
Total wec
-
.....
LM -
.
1
Sono
6000
5000
2000
.000
CHART XX.
Rabbit No. 108 showing total leucocyte counts and absolute values of polymorphonuclear, large and small mononuclear leuco- cytes for entire period of observation.
Digitized by
ma's
476 81
₦76 6/
June /
was
9 am
mai
mas
6/
of
1
.
5000
1000
2000
June 16 9 am
meds
June 29 9am
Jame 29 9am
wed's
268
JOHNS HOPKINS HOSPITAL BULLETIN.
[No. 15
Rabbit No. 107-Cont.
Total
Pmn
Lm
Sm
Eog
Mst % Total
June
5180
43.5 2253
6.0
311
45.5 2357
1.5
78
8.5
181
1 p. m.
5940
88.5 2287
48.5
3375
5.5
383
37.5
2610
0.5
85
8.0
557
2, 9 a. m.
4040
85.5
1434
842
8.0
421
70.0
8682
0.5
26
5.5
289
4, 9 a. m.
5000
41.0 2050
4.5
225
49.0 2450
..
5.5
275
June 16, 9 a. m.
5200
88.0 1716
5.0
260
55.5 2886
0.5
26
6.0
312
(Inoc. 2.5 cc. serum intravenously.)
1 p. m.
7140
80.0
2142
4.0
286
62.5 4462
0.5
36
3.0
214
5 p. m.
6860
37.0
2538
6.5
446
46.0
8156
10.5
720
17, 9 a. m.
5020
84.0
1707
87.5 1283
45.5 1756
11.0
425
84.0 1312
9.5
367
June 29, 9 a. m. 4300
(Inoc. 0.01 cc. serum intradermally.)
1 p. m.
1800
6780
49.0
3322
6.0
407
39.0 2644
6.0
407
July
1, 9 a. m.
4960
52.0 2579
85.5
4170
7.0
290
53.0 2194
1.0
41
8.5
145
Rabbit No. 108 May 23, 9 a. m.
3600 7920
4700
51.5
2420
1294
1733
1296
6.0
190
48.5
1532
63
2.5
79
June 1, 9 a. m.
4420
89.2
1654
10.8
456
273
739
418
175
47.0
2059
197
4, 9 a. m.
8120
48.0
1841
8.0
250
46.0
1485
8.0
94
June 16, 9 a. m.
3740
80.0 1122
8.5
818
59.0 2206
0.5
19
2.0
75
(Inoc. 5.0 cc. serum intravenously.)
1 p. m.
6340
58.0
3677
6.5
412
751
204
5.5
183
47.5
1577
8.5
116
19, 9 a. m.
4080
46.5 1897
45.0 1859
2.5
75
50.0 1510
0.5
15
2.0
60
(Inoc. 0.01 cc. serum intradermally.)
1 p. m.
4660
43.0 2004
2444
10.0
404
8,0
98
13.5
440
0.5
16
5.0
163
2, 9 a. m.
8620
25.5
923
6.5
285
64.0 2317
36
3.0
108
Chart XVI shows the total leucocyte counts for the entire series plotted out so that a general idea of the variations which have taken place can be gotten at a glance.
One sees that the leucocytes underwent as great changes in number during the two series of observations before inocula- tion as they did after inoculation. On May 23, June 1, 16 and 29, days on which counts were made at 9 a. m., 1 p. m. and 5 p. m., it is to be observed that the 1 p. m. count, or the 5 p. m. count, or both, are higher than the 9 a. m. count. This we ascribe to a digestive leucocytosis as in the preceding series, the animals being fed at 10 a. m. each day. We are unable to see that the intravenous injection of normal horse serum exerted any influence on the total number of leuco- cytes. It is possible that the depression of the digestive leu- cocytosis which is observable on June 29 may have been due to the intradermal inoculation.
Charts XVII, XVIII, XIX and XX show the total leuco- cytes and absolute values of the differential counts plotted out for rabbits Nos. 105, 106, 107 and 108, respectively. They illustrate very well the observation made in connection with the preceding series; namely, that both the polymorphonu- clear and small mononuclear leucocytes participate in an in- crease in the total number of leucocytes, but in this series the small mononuclears, as a rule, participate to a greater extent than do the polymorphonuclears .* It is possible that the ex-
* These results are in entire agreement with those obtained in a fourth series of counts which are not included in this report on account of lack of space.
ception to this rule observable in Charts XVIII and IT June 16, 1 p. m., where we see a relatively greater incha. in the polymorphonuclear cells may have been due to the : travenous injection of normal horse serum, but inasmoxb : similar exceptions are to be seen elsewhere in the charts res no horse serum was inoculated, it would hardly be sale : draw conclusions from these two instances.
Curves showing the total number of polymorphonne's leucocytes for the entire series were superimposed for pure of comparison and the same was done for the small to nuclears, but as the injection of serum seems to have bel entirely without influence on either of these elements z charts are omitted.
SUMMARY.
We have thought it worth while to publish these studie full, giving the detailed counts for each series, since te represent a large number of carefully made observations ez the data may be useful to others who are attempting to day conclusions from variations of the leucocyte counts in rats
We feel justified in saying that the normal variation in t. total leucocyte count taken at the same hour each day : rabbits kept as nearly as possible under constant condi: is very great, at times reaching nearly 100 per cent.
We have observed quite regularly a diurnal cycle in ri we are inclined to ascribe the increase in the number of le .: cytes to the influence of digestion.
The relative and absolute values of the differential cnc vary under normal conditions from day to day and at differa hours of the same day.
We observed that an increase in the total leucocyte (: - was participated in, as a rule, by both the polymorphonxcre and small mononuclear leucocytes, but usually by the latk" a greater extent than by the former.
The relative values (percentage values) of the poloz phonuclear and small mononuclear leucocytes bear a recipro relation to each other; that is, an increase in the percerias of one is accompanied by a decrease in the percentage of : other.
We were unable to observe that the intravenous injecting normal horse serum in doses varying from 0.1 ce. to 5 (c. the intradermal injection of 0.01 cc. normal horse serum : either sensitized or non-sensitized rabbits had any definite constant influence on the total or differential leucocyte cozza three hours or longer after injection.
We have a few observations, not included in this ps?" where counts were made at short intervals (5-10 minutes) cfa the intravenous injection of large amounts (10 ce.) nocz- horse serum into rabbits. These indicate, as far as they : that shortly after injection the total count undergoes a sz: decline with a marked relative increase of polymorph: clear and decrease of small mononuclear leucocytes. T :- observations, however, were few in number and we hesita: draw conclusions from them.
20
1.5 60
July
1, 9 a. m.
2760
25.5
704
9.5
262
61.0 1688
3.5 0.5
163
2.5
116
5 p. m.
80, 9 a. m.
3260
78.0 2543
1919
8.5
0.5
25
1.5
74
8, 9 a. m.
4380
44.5
1949
5.6
436
188
43.0
2021
1.5
71
..
..
4.5
204
25, 9 a. m.
4280
40.5
6.0
257
51.5 2204
21
1.5
64
26, 9 a. m.
3160
1 p. m.
6840
49.5
8886
4.0
8.5
181
2.0
174
2, 9 a. m.
4920
89.0
45.5 1429
1429
0.5
16
2.0
62
18, 9 a. m.
3820
43.5 1444
6.5
265
43.0
1754
1.0
32
8.5
589
5 p. m.
6260
49.5
8099
12.0
26.5 86.0
2254
2.5
156
17, 9 a. m.
8140
35.0 2824
8.0
646
248
40.0
1984
3.0
149
2, 9 a. m.
4140
40.0 1440
4.0
144
54.0 1944
1.0 0.5
86
1.0 2.5
36
1 p. m.
43.5
3445
3801
40
198
24, 9 a. m.
4540
28.5
12.5
56
54.5
2484
0.5 2.0
0.8 1.0 1.5
33 68
2.4 1.0
68
5 p. m.
8700
44.5
3871
43.5
50.5 2484
..
: :
8.0
122
June 29, 9 a. m.
3020
4040
60.5
10.5
624
49.0
2910
2.0
119
5 p. m.
1.0
40
8.5
343
3, 9 a. m.
5260
16.0
7.0
851
7.0
239
51.5 1761
..
4.0
187
19, 9 a. m.
3860
35.0 1505
7.5
823
51.0 2198
6.5
279
5 p. m.
30, 9 a. m.
8070
4196
0.5
4.5
883
:
..
..
58.0 2661
6.0
901
18, 9 a. m.
3420
4.5
182
50.5
2040
% Total
Leucocytes % Total
Total % Total
1, 9 a. m.
0.5 1.0
14
3.5 96
6.5
4.5
210
46.5 2167 27.5 1111
46.8 44.5
1975 8044 $784
1680
0.5
..
-
Digitized by
4.0
4.5
5.0
4.0
41.0
101
5 p. m.
48.0
45.5
52.0
6960
THE TUBERCLE BACILLUS. By W. W. BOARDMAN, M. D.
(From the Research Laboratory, The Phipps Tuberculosis Dispensary, The Johns Hopkins Hospital.)
In absolute diagnosis of pulmonary tuberculosis can be ed upon one, and only one finding-the demonstration of ercle bacilli in the sputum. Yet in a review of the reports arious tuberculosis dispensaries and sanatoria we find that he cases clinically diagnosed as definite pulmonary tuber- sis, the sputum is negative for tubercle bacilli in from 46 per cent. Are we to conclude then, that in from 8 to per cent of our clinically positive cases, there are no tubercle illi in the sputum, and that these cases therefore are ligible factors in the spread of the disease; or that in a ;e proportion of these cases we fail to demonstrate tu- le bacilli, not because of their absence, but as a result of imperfect methods of sputum examination, or our too less use of these methods ?
.s to the first possibility; clinically positive cases without ercle bacilli in the sputum do exist among the arrested s and possibly in the very early stages of the disease, but 'e is no doubt that the number of cases so classed at the sent time is too large.
'he second possibility seems a more probable explanation. fail to find tubercle bacilli in the sputum of many clini- y positive cases, since (a) the specimen examined may con- i no tubercle bacilli, being merely mucus from the upper passages; or (b) if the tubercle bacilli are present in but Il numbers, they may be overlooked, even in the most care- examination by the ordinary methods. This point is well strated by the following report by Goerres. In examining ' smears from each of 296 specimens-96 were demon- ted to contain tubercle bacilli by the first smear, 24 more the second, 9 more by the third, and 5 more by the th, and yet of the remaining 162 apparently negative a, 28 were demonstrated to contain tubercle bacilli by the formin method. Finally (c) as to the careless use of present methods, it is evident, that if some cases escape ction even with repeated smear examinations, many may pe detection when but a single smear is made.
hat then is the solution of the problem? It is evident we must first obtain a specimen from the tuberculous n; again we must examine several smears from the same imen; and lastly we must examine repeated specimens. observance of these rules calls for the expenditure of i time and energy, and in return, although the results fairly accurate, they still are far from perfect.
more nearly absolute method, first proposed by Biedert ' slightly modified by subsequent investigators (Goerres ') long been recognized as a valuable procedure, but the us technique has prevented its use except in a compara- y few doubtful cases. Briefly, the method is as follows : large quantity of sputum is well diluted with distilled : and 0.2 per cent sodium hydroxide is added, the material
heated, stirring meanwhile, until a homogeneous solution re- sults. It is then neutralized with acetic acid, and double the amount of 96 per cent alcohol added. It is then allowed to settle, the supernatant fluid is poured off and the sediment examined in the usual way. This method facilitates the find- ing of the bacilli by concentrating them in the sediment.
Following Biedert's first publication various agents were recommended for dissolving the mucus, such as hydrogen peroxide, carbolic acid, and pancreatic ferment, but none of these give so satisfactory results as the weak solution of sodium hydroxide with heat.
Thus matters stood until in the latter part of 1908, when Uhlenhuth ' reported the results of his extensive researches with antiformin, a preparation much used by brewers in the disinfection of their fermentation vats and pipes.
Antiformin is a clear yellowish liquid of high specific gravity, possessing a faint chlorine odor and a decided soapy feel. According to Uhlenhuth it is composed of sodium hy- droxide 7.5 per cent with sodium hypochlorite in such amount that 100 grams of antiformin liberates five and three-tenths grams of chlorine gas. After months of standing in the laboratory there is no appreciable deterioration.
As a result of his researches Uhlenhuth recommends anti- formin as a disinfectant of the first rank, surpassing in many respects carbolic acid and bichloride of mercury. Its effi- ciency depends upon an intensive oxidation process, this being so marked that practically all organic matter except hair, wax, fat and cellulose, is rapidly brought into solution-the rate and completeness depending upon the strength of the solution used and the temperature at which the reaction occurs.
Upon adding a solution of antiformin to sputum, there is a marked liberation of gas. The sputum rises to the top of the mixture and rapidly dissolves, the result being a practically homogeneous solution, varying in color from yellowish-brown to a pale yellow, with but a small amount of flocculent sedi- ment. The consistency of the resulting liquid varies directly with the consistency of the original specimen. The sediment varies in color from white to a dirty gray and consists of a finely granular detritus, fat needles, dust particles, cellulose fiber and any acid-fast organisms which may have been pre- sent in the original specimen.
A similar reaction occurs upon adding antiformin to finely divided animal tissues, pus, feces, suspensions of bacteria, etc. Uhlenhuth concludes that with the exception of one group of bacteria, all bacteria, protozoa, spirochætæ, and trypano- soma are dissolved in 2 to 5 per cent antiformin solution in from two and one-half to five minutes, the majority of them undergoing almost instantaneous solution. The organisms of the acid-fast group are the only ones resisting the dissolving
Digitized by
270
JOHNS HOPKINS HOSPITAL BULLETIN.
[Se
action of the antiformin solutions and this is explained by the fact that they are enveloped in a waxy capsule. Further work by Uhlenhuth and others revealed the important fact that tubercle bacilli, as shown by animal experimentation, are rapidly killed by antiformin only in 50 per cent or stronger solutions; that they retain their virulence for 12 days in 8 per cent solutions and for 4 days in 20 per cent solutions; finally that their staining properties are practically unaffected by exposure to 50 per cent antiformin for weeks.
We therefore have in antiformin an agent capable of rapidly and completely reducing sputum to a practically homogeneous solution without altering the staining qualities or viability of the contained tubercle bacilli. Several methods have been proposed for the recovery of the bacilli from these solutions, thereby making the procedure of practical value.
Uhlenhuth recommends simple sedimentation, his method for sputum examination being briefly as follows :
1. 15-30 cc. of sputum in a conical settling glass.
2. Add from three to five times the volume of water.
3. Add sufficient antiformin to make a 15 per cent solution.
4. Stir thoroughly and allow to digest and settle com- pletely.
5. Pour off supernatant fluid.
6. Wash sediment with sterile water and allow sediment to reaccumulate.
7. Pour off supernatant fluid; smear sediment on slide and examine in usual manner.
The washing was found necessary since the presence of any considerable quantity of antiformin prevented firm fixing of the sediment to the slide.
Hüne ' uses both the gravity sedimentation and the cen- trifuge. He proposes two distinct methods, as follows : *
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