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The histology and physiology of normal pigs' blood
GENERAL ARTICLES
We have all the primary conditions causing a discoloration of the bile and yellow infiltration of the system. Noticing that most of the animals are immune against this disease, we may explain it in the same way as redwater, that is, animals born in South Africa become immune, whereas those imported contract it. Therefore the disease caused by piroplasma mutans is not so frequent amongst animals bred on the veldt in South Africa as in animals bred in stables or imported. This view has lately been borne out by some observations in practice, where gall-sickness was reported and microscopical examinations proved the presence of piroplasma mutans. The presence of piroplasma mutans in the blood of animals is of the utmost importance from a diagnostic point of view, because we diagnose East Coast fever by the presence of piroplasma parvum, and, as may be seen from the accompanying Plate, there is hardly any difference between these two piroplasms. The former causes a disease which ends fatally; it rapidly increases in numbers as the disease advances, and this is one of the differential points which we apply in practice. It sometimes happens, however, that animals in an infected area die before piroplasma parvum has developed to any extent, and in such cases the diagnosis must remain doubtful. A short time back Robertson of Cape Colony made the statement that East Coast fever may run its course with the total absence of piroplasma parvum, or perhaps only with the presence of a very small number of these parasites. We are not able to distinguish such cases from piroplasma mutans during life by microscopical examination, and a post-mortem examination is the only way to form a correct diagnosis. But, again, piroplasma parvum does not always produce the typical lesions, so that the diagnosis has to be left an open one. It is therefore sometimes most difficult, if not impossible, to distinguish between piroplasma parvum and piroplasma mutans.
THE HISTOLOGY AND PHYSIOLOGY OF NORMAL PIGS' BLOOD.
By 'WARD GILTNER, D.V.M., Assistant Veterinarian, Alabama Polytechnic Institute, U.S.A. WITHIN the last few years the study of the blood of the domesticated animals has received no little attention from certain investigators. In fact we now have authoritative records on the normal condition of the blood of the horse, dog, and cow. Swine have not, however, received the attention of h
Journal of Comparative Pathology and Therapeutics.
Plate J
..
""
'P. mu tans
P big eminum
P. parvum
~.
.....
""
OeuI N'? 6 T.engtb of tube 180 III m . ObJe ct· '12 I:mmers N. Ap J. 30.
GENERAL ARTICLES.
resume of the data on the blood of the pig. It is especially interesting in this connection for purposes of comparison or contrast. The red corpuscles of the blood of the pig have an average diameter of 6 microns, the size varying from 5'28 microns to 7"9 microns (Bethe 1). The number of red corpuscles is given as 5,440,000 (Stoltzing 3) and 6,960,000 (Bethe 1) per cmm. The specific gravity is given by Sussdorf 4 as about 1060. Drake 6 gives the varieties of leucocytes for fifteen normals as: lymphocytes, 33 to 77 per cent.-average, 56'4 per cent.; polynuclears, 18 to 66 per cent.-average, 38'46 per cent.; eosinophiles, I to 12 per cent.-average, 5'13 per cent. Red Corp".'cl£s. ! Leucocytes. Per cmm.!
Hb. I 81'. gr. Per cent. \
Size of Red Corpuscles.
,
Author.
------1--1----'----,
6,960,000
I
7840
-
II
-
I
I
i 5'28 to 7"9
microns 6 microns
I Bethe. 1 ' I
Gulliver.2 Stoltzing. 3
1060 8,668,200
6 microns
'Sussdorf.4 Wendelstadt' und Bleibtreu.
It must be confessed that the above table is a mere skeleton and evidently needs filling in. A glance at the various columns shows that our knowledge of the pig's blood is very meagre and at the same time unsatisfactory. For instance, the column for Hb. contains absolutely no figures, while the results for the red cells do not possess the uniformity demanded by accuracy. It is safe to presume that the technique employed by the earlier investigators was rather crude, which may account for low' counts of St6ltzing and Bethe. The object of the following data is to confirm or negative the results and in a measure to supplement them. No delicate histological methods have been employed, and the results are far from exhaustive, but are merely the outcome of a careful preliminary investigation covering a short period of time and conducted with the simplest possible technique, such as may be adopted by the busiest investigator or practitioner. Only a few measurements of the red cells have been made, but those give an average diameter for the red cell of about 6 microns. The specific gravity as deter1 Bethe, Martin: Beitn\ge zur Kentniss der Zahl- und Massverhtiltuisse der rothen Blutk(irperchen. Schalbes'" l\Iorpholog. Arbeit," Bd. VII., Heft 2, p_ 207, 1092. 2 Gulliver, Geo. : Observatious on the Sizes and Shapes of the Red Corpuscles of Vertebrates, etc. "Proceed. of the Zoological Soc.," London, 1875, pp. 169-176. :l Stbltzing, 'V.: Uber Zahlung Blutkorperchen. "lnaug. Diss.," Marburg, 1856. 4 Sussdorf, M.: Blut und Blutbewegung. Ellenberger's" Handbuch der vergleichenden Physiol. der Haussaugethiere," Bd. II., Thiel 1, 1890, pp. 163-216. J 'Vendelstadt, H., und Bleibtreu, L. : Bestimmung des volumens und des Stickstoffgehaltes des einzelnen rothen Blutk6rperchens im Pferde und Schweine Blut. "Archiv. fur gesamm. Physiol.," Bd. LII., 1892, p. 332. 6 Drake, A. K.: Trichinosis. "Jour. of l\Ied. Research," Vol. III. (8), June 1902, p. 255.
20
G ENERAL ARTICLES.
mined by Hammerschlag's method was found not to differ materially from that recorded by Sussdorf. The method of securin g the blood was made as simple as possible and at the same time consistent with accuracy. The subject for examination was cau ght in a small pen without any avoidable excitement that might alter the normal blood count, and secured on ,a table with no other restraint than an assistant holding the legs, thus leaving the ear~ or tail free for the withdrawal of the blood. In the case of large hogs they can be secured by letting them step in a noose in a rope and then throwing them on the back, when resistance is speedily overcome. No trouble was experienced on account of resistance by the animal while drawing the blood; in this respect the pig b eing preferred by the writer to horse, cow, dog, or sheep for purposes of blood examination. In the first cases the ear was carefully washed and dried, but later this preliminary step was aband oned. After clipping away the long hairs and wiping the ear with a dry cloth, an incision was made in either posterior or anterior border, or a puncture was made in a vascular part of the ear with a sharp scalpel. By this procedure a good flow of cutaneous blood was secured, the first few drops being wiped away and the following drops absorbed by the unsized paper used in Tallquist's method of determining the h~moglobin percentage. This was the only method employed in estimating the h~mog lobin, and, while g reat accuracy cannot be claimed (Cabot allows an error of 10 per cent. for it), it has the advantage of simplicity and availability in all cases under all circumstances. Smears were mad e o n specially cleaned slides, three or four being made in each case. The blood for estimating the erythrocytes and leucocytes was drawn in the same pipette, using Toisson's diluting fluid, and in every case securing a dilution of I to 100, the estimation being made by the Thoma-Zeiss h~matocytometer with the Zappert-Ewin g ruling. An objection might be made to the lack of proper preparation of the field for making the incision, but the method adopted was not attended by any bad results, such as the introduction of epithelial cells or fat-drops of sebum in excess to interfere with the leucocy te count, a nd it has the advantage of not introducing the error of increasin g the number of leucocytes by manipulation of the part. In estimating the number of red cells 100 squares were counted, on each of two slides, in most cases, and, when the results were not close, other slides were prepared until what was thought to be a fair average was secured. The whol e ruled space, or 9 to 10· sq. mm., was counted on two or more slides for the white cells. The differential count of leucocytes was macle in all cases from slides stained with Jenner's stain as modified by Dr C. W. Hastin g, which was found to be exceedingly satisfactory. Numerous smears stained by Wright's method and by Ehrlich's tri-acid stain revealed little not brought out by Jenner's. A total of at least 500 leucocytes was counted in each case, and any unusual results were confirmed or corrected by examination of additional slides, even in some cases by securing more films from the subject in question. While Drake apparently recognises only three varieties of leu cocytes, no difficulty is experienced in distinguishing five varieties, unless there be some hesitancy about separating the lymphocytes and large mononuclears. Certainly the mast cells cannot be over-
GENERAL ARTICLES.
21
looked when stained by Jenner's stain. The lymphocytes are slightly larger than the red cells, generally spherical in shape, from 7'4 to 10 microns in diameter, and averaging about S'5 microns. They have a blue nucleus which occupies nearly the whole cell, showing in most cases a crescentic-shaped, darker or lighter blue portion of the cell body at the periphery. Much larger cells, I I to 14 microns in diameter, are found in far fewer numbers, with round nuclei, occupying relatively less of the cell space, while the more typical large mononuclear leucocytes are similar in size and have a mediumsized nucleus, light blue and bean-shaped, and show a large portion of the cell body stained a different shade of blue. The polynuclears closely resem ble those of the horse in size (10 to I4 microns in diameter) and in morphology of the nucleus, which is exceedingly polymorphous, and takes the blue stain; but the cell body is, in most of the specimens, barely visible unless the procedure in staining is very careful. By carefully timing the staining, washing, and rapidly drying in the air, the cytoplasm is seen to be a faintly pinkish colour with a circular outline to the cell body. The eosinophiles are comparable to the polynuclears in size or slightly larger, and have a bi-Iobed nucleus, the two parts of which are connected by a thick band and take the basic stain. The cell body is granular, but the granules are not nearly so large and distinct as those found in the eosinophile of the horse, but are more numerous, estimated at 100 more or less, and have a strong affinity for the eosin stain. The mast cells are about the same size as the preceding and have a similar shaped nucleus, but the cell body possesses granules of a smaller size, more distinct in outline, and of a purple colour. The granules lie both in the cytoplasm surrounding the nucleus and in a position superposed to the nucleus. In hurriedly made smears the erythrocytes are badly crenated, more especially if the smear is thick; otherwise they are circular in outline, and differ in no material respect from those of the horse or man. Many or most preparations show more or less numerous clusters of small bodies, I to 2 microns in diameter, faintly blue in colour, seeming not to take the stain diffusely. These have no particular relation to either the red or white cells as to location and are presumably blood plates. However, Cabot says the blood plates take the eosin stain, while Ewing admits that they take either the acid or basic stain. In all, the blood of twenty-four different pigs has been examined and the results tabulated. All the cases were considered normal, being neither exceedingly poor nor excessively fat and possessed of the characteristic appetite, varying in size, colour, breed, sex, and source, though mostly from the surrounding farms. Intestinal and other parasites are common in swine in the locality of the Institute, but only a post-mortem examination will reveal to what extent th}! cases examined are infested. An accurate knowledge of the effect of these parasitisms on the blood is wanting. Numbers 13 to IS had recently been weaned and were in good condition, and, it will be noticed, show a very close comparison in results of the blood counts. Pig 19 was the only pure white one in the lot and differed materially from the others in conformation, having an undesirable length of nose and legs; yet his lymphocyte count shows a large percentage, far
GENERAL ARTICLES.
22
larger than in many of the much better nourished animals. From this it seems rather difficult to point any constant relation between the percentage of the different varieties of Jeucocytes and the nutrition of the animal, although it would seem that the lymphocytes might increase in fatter animals. In Case 9, the only black and white spotted pig, there is an unaccounted-for increase in the eosinophiles ; consequently, they are not considered in figuring the general averages. The averages of the different varieties of leucocytes are calculated both by working out the percentages from the average number of leucocytes (I9,oOO) and by using the results in the different columns. These results compare favourably. All of the too few standard works on hcematology and blood have been consulted, and only a few references of the more important articles immediately connected with the subject in hand are included in the references. This work has been done under the direction of Dr C. A. Cary, whose constant advice and suggestions have materially assisted in accomplishing satisfactory results. Thanks are due to Dr Cary's assistant, Mr C. T. Butler, B.S., for his intelligent assistance and co-operation, both in securing the blood and in counting the erythrocytes. TABLE
I. -
Ko.
Se.c.
Date 0/ TakttlY.
Age.
II". Pel' {'lilt.
EI·lIt1t"ocl'le.~.
Pel'
Clnllt.
Leucoc,1Ite-<,
-
Reillur/.s.
Pel' emllt.
- - - - - - - - - - - - ------ - - -- - - - - - - - 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.
17-
18. 1H. 20. 21. 22. 23. 24.
Max.
l\1in. Av.
C. m. S. f. S. f. S. f. S. f.
S. f. S. f.
11/13/06 11/Hi/06 11/16/06 11/16/06 11/20/06 n/21/06 11/22/06 11/23/06 11/26/06 11/27/06 11/28/06 11/29/06 12/4/06 12/3/06 12/4/06 12/5/06 12/6/06 12/7/06 12/10/06 12/11/06 12/11/06 12/12/0H 12/12/06 12/13/06
14m. 10 F.
-
M. F.
C.m.
l\1. 111. l\1. 111. 111. 1II. F.
C. m. F.
l\I.
-
-
-1 mo. 90 4 mo. 100 4mo. 85 4 mo. 95 4 mo. 90 4 mo. 00 4 mo. 90 4 mo. 80 5 mo. 80 3 mo. 85 85 3 mo. 95 5 mo. 2'5 mo. 90 2'5 mo. 85 2';' mo. 85 2'5 mo. 85 2',1 mo. 85 2'5 mo. 85 Hmo. HO G mo. 85 6mo. 80 100 6mo. G mo. 85 6 mo. 90
8,200,000 7, 72n, 000 7,750.000 7,180,000 8,700,000 8,050,000 \ 8,800,000
18,000 22,000 18,300 10,000 20,000 14,500 19,000 22,000 1R,000 16,000 21,500 17,000 22,000 12,000 16,000 20,000 21,500 17,000 9.000 25,000 19,000 16,500 23,500 24,000
100 80 88
8,800,000 6,800,000 8,450,000
25,000 9,nOO 19,000
6 mo. 2':; mo. 4 mo.
7,500,000 8,000,000 8,400,000 8,400,000 7,600,000 7,300,000 8,000,000 7,000,000 7,100,000 6,800,000 7,700,000 R,OOO,OOO 7,900,000 8,400,000 8,600,000 8,800,000 8,~80,000
Black, in fair condition.
"
" " " " Bl~ck. " Bl~ck and white,
" " "
under.ized Black and white, spotted. Black, runt rnptured. Small, black. Black, good condition. Black, just weaned.
" "
"
"
" " " " "
Ail white. Black. Black, undersizf'a. Black.
B{~ck, large, well fed.
--
I
Note.-C, castrated; S, spayed; lIf, Male; F, female.
---
23
GENERAL ARTICLES,
TABLE
II,
Differential Count of Lellcoc)'tes, Ly IIIphocytes. No,
Le1!c()cptt!x.
Per
Lar{fe 1lfononucleal',
Polynucl~ar.
-----
cml/~.
I
Mast Cells,
Eo,·"finopltiles.
I cent. 1''''1 1'.0.. 1cent. Pel' I
Pel' I lio. . cent. No. I No. - -- - -- - - -- - - - - - - - - - - - - - - - - -
Pel' cent.
1. 2, 3, 4, 5, Ii. 7. 8.
fl. 1O, 11, 12. 1:~,
14. 15,
]6,
17, IS, 19, 2O, 21. 22, 2:t 24, l\Ia", lIIin, Av, 2nd Av,
I I
No.
Pel' cent.
18,000 22,000 18,300 10,001) 20,000 14,500 19,000 22,000 18,000 16,000 21,500 17,000 22,000 12,000 16,000 20,000 21,500 17,000 9,500 25,000 19,600 16,500 23,500 24,500
30 32 52'6 37 46 45'4 44'4 48'6 43 60 44 47 60 6ii'8 51'4 79'8 57'4 45 68'S 54 67"4
5,400 7,040 9,626 3,700 9,200 6,583 l',436 10,(;92 7,740 9,600 9,460 7,990 13,200 7,896 8,224 15,960 ]2,341 7,650 6,536 13,500 13,210'4 ,Ill 8,085 12,4;)5 53 50'6 12,397
6'4 1,152 8 1,760 1,09H 6 1,000 10 2'8 ' 560 ;; 725. 3'(j Ij84 4·t) 1,012 9'8 1,764 4'2 672 4'8 1 1,033 5'4 , 918 2'0 57:! 1'1) ! 197 4 640 400 2 2 430 2'4 400 152 1'6 4 1,000 '8 1568 2'6 ! 429 ! 70,1 :\ 2'8 686
25,000 9,500 HI, 000
79'S 15,960 30 3,700 51'6 9,400 !l,890
10
-
'8 I 4'6 I --
1,760 1Ii2 715 874
10,800 2'2 12,540 2 6,954 1'2 5,000 no 1'8 45'6 9,120 5 44 6,380 4 44 8,360 6'6 38 8,360 8 3,780 25'S 21 :~,360 21 9'2 34'S 7,480 11 39'2 6,664 6'4 34'2 7,524 3 26'2 3,144 3'S 6,400 40 4 13 2,600 4'S 5'4 ~5 7,525 47'4 S,058 5 2,5 2,37n 4 33 8,250 8 23'(j 4,625'6 7'8 40 6,660 6'6 36'4 8,554 7 38'8 9,50G 6'4 60 12,540 11 1;\ 2,375 1'2 6,820 5'2 37 -7,030 60 57 38
396 440 219'5 180 1,000 580 1,254 1,760 4,644 1,472 2,3/i5 1,088 660 466 640 960 1,161 8.'iO 380 2,000 1,528'8 1,089 1,645 1,568
1'4 1 2'2 1'2 '6 l'Ii 1'4 'S '4 5'6 5'4 2 '2 'S '6 '4 '2 '2 '6 1 '4 1'8 '6 1'4
255 220 402'5 120 120 232 266 176 72 S96 1,162 340 44 96 96 80 43 34 57 250 78'4 297 141 343
2,360 180 1,050 988
5'6 '2 1'3
1,162 34 245 247
-
INDIAN EQUINE PIROPLASMOSIS.
By
CAPT,
A.
J,
WILL1AMS,
F,R,C.V,S" A.V,c.
History,
THIS disease has been known in India for a considerable time under the name of bilious fever, and in South Africa an identical disease was commonly known as biliary fever, According to Hutcheon, it was described by veterinary surgeons in Natal, as far back as 1883, as the biliary form of horse-sickness and as anthrax fever; Hutcheon also states that, although it strongly resembled horse-sickness, he was convinced that it was of a distinct origin and amenable to different treatment, • In the same article Bowhill 5tates that, as far as he was able to ascertain, Hutcheon's conclusions are accepted by all recent writers except Edington, who divides horse-sickness into two forms, one of which he describes as a non-virulent form with a malarial type of fever and in which parasites are found, Theiler admits no co-relation between horse-sickness and equine
We have all the primary conditions causing a discoloration of the bile and yellow infiltration of the system. Noticing that most of the animals are immune against this disease, we may explain it in the same way as redwater, that is, animals born in South Africa become immune, whereas those imported contract it. Therefore the disease caused by piroplasma mutans is not so frequent amongst animals bred on the veldt in South Africa as in animals bred in stables or imported. This view has lately been borne out by some observations in practice, where gall-sickness was reported and microscopical examinations proved the presence of piroplasma mutans. The presence of piroplasma mutans in the blood of animals is of the utmost importance from a diagnostic point of view, because we diagnose East Coast fever by the presence of piroplasma parvum, and, as may be seen from the accompanying Plate, there is hardly any difference between these two piroplasms. The former causes a disease which ends fatally; it rapidly increases in numbers as the disease advances, and this is one of the differential points which we apply in practice. It sometimes happens, however, that animals in an infected area die before piroplasma parvum has developed to any extent, and in such cases the diagnosis must remain doubtful. A short time back Robertson of Cape Colony made the statement that East Coast fever may run its course with the total absence of piroplasma parvum, or perhaps only with the presence of a very small number of these parasites. We are not able to distinguish such cases from piroplasma mutans during life by microscopical examination, and a post-mortem examination is the only way to form a correct diagnosis. But, again, piroplasma parvum does not always produce the typical lesions, so that the diagnosis has to be left an open one. It is therefore sometimes most difficult, if not impossible, to distinguish between piroplasma parvum and piroplasma mutans.
THE HISTOLOGY AND PHYSIOLOGY OF NORMAL PIGS' BLOOD.
By 'WARD GILTNER, D.V.M., Assistant Veterinarian, Alabama Polytechnic Institute, U.S.A. WITHIN the last few years the study of the blood of the domesticated animals has received no little attention from certain investigators. In fact we now have authoritative records on the normal condition of the blood of the horse, dog, and cow. Swine have not, however, received the attention of h
Journal of Comparative Pathology and Therapeutics.
Plate J
..
""
'P. mu tans
P big eminum
P. parvum
~.
.....
""
OeuI N'? 6 T.engtb of tube 180 III m . ObJe ct· '12 I:mmers N. Ap J. 30.
GENERAL ARTICLES.
resume of the data on the blood of the pig. It is especially interesting in this connection for purposes of comparison or contrast. The red corpuscles of the blood of the pig have an average diameter of 6 microns, the size varying from 5'28 microns to 7"9 microns (Bethe 1). The number of red corpuscles is given as 5,440,000 (Stoltzing 3) and 6,960,000 (Bethe 1) per cmm. The specific gravity is given by Sussdorf 4 as about 1060. Drake 6 gives the varieties of leucocytes for fifteen normals as: lymphocytes, 33 to 77 per cent.-average, 56'4 per cent.; polynuclears, 18 to 66 per cent.-average, 38'46 per cent.; eosinophiles, I to 12 per cent.-average, 5'13 per cent. Red Corp".'cl£s. ! Leucocytes. Per cmm.!
Hb. I 81'. gr. Per cent. \
Size of Red Corpuscles.
,
Author.
------1--1----'----,
6,960,000
I
7840
-
II
-
I
I
i 5'28 to 7"9
microns 6 microns
I Bethe. 1 ' I
Gulliver.2 Stoltzing. 3
1060 8,668,200
6 microns
'Sussdorf.4 Wendelstadt' und Bleibtreu.
It must be confessed that the above table is a mere skeleton and evidently needs filling in. A glance at the various columns shows that our knowledge of the pig's blood is very meagre and at the same time unsatisfactory. For instance, the column for Hb. contains absolutely no figures, while the results for the red cells do not possess the uniformity demanded by accuracy. It is safe to presume that the technique employed by the earlier investigators was rather crude, which may account for low' counts of St6ltzing and Bethe. The object of the following data is to confirm or negative the results and in a measure to supplement them. No delicate histological methods have been employed, and the results are far from exhaustive, but are merely the outcome of a careful preliminary investigation covering a short period of time and conducted with the simplest possible technique, such as may be adopted by the busiest investigator or practitioner. Only a few measurements of the red cells have been made, but those give an average diameter for the red cell of about 6 microns. The specific gravity as deter1 Bethe, Martin: Beitn\ge zur Kentniss der Zahl- und Massverhtiltuisse der rothen Blutk(irperchen. Schalbes'" l\Iorpholog. Arbeit," Bd. VII., Heft 2, p_ 207, 1092. 2 Gulliver, Geo. : Observatious on the Sizes and Shapes of the Red Corpuscles of Vertebrates, etc. "Proceed. of the Zoological Soc.," London, 1875, pp. 169-176. :l Stbltzing, 'V.: Uber Zahlung Blutkorperchen. "lnaug. Diss.," Marburg, 1856. 4 Sussdorf, M.: Blut und Blutbewegung. Ellenberger's" Handbuch der vergleichenden Physiol. der Haussaugethiere," Bd. II., Thiel 1, 1890, pp. 163-216. J 'Vendelstadt, H., und Bleibtreu, L. : Bestimmung des volumens und des Stickstoffgehaltes des einzelnen rothen Blutk6rperchens im Pferde und Schweine Blut. "Archiv. fur gesamm. Physiol.," Bd. LII., 1892, p. 332. 6 Drake, A. K.: Trichinosis. "Jour. of l\Ied. Research," Vol. III. (8), June 1902, p. 255.
20
G ENERAL ARTICLES.
mined by Hammerschlag's method was found not to differ materially from that recorded by Sussdorf. The method of securin g the blood was made as simple as possible and at the same time consistent with accuracy. The subject for examination was cau ght in a small pen without any avoidable excitement that might alter the normal blood count, and secured on ,a table with no other restraint than an assistant holding the legs, thus leaving the ear~ or tail free for the withdrawal of the blood. In the case of large hogs they can be secured by letting them step in a noose in a rope and then throwing them on the back, when resistance is speedily overcome. No trouble was experienced on account of resistance by the animal while drawing the blood; in this respect the pig b eing preferred by the writer to horse, cow, dog, or sheep for purposes of blood examination. In the first cases the ear was carefully washed and dried, but later this preliminary step was aband oned. After clipping away the long hairs and wiping the ear with a dry cloth, an incision was made in either posterior or anterior border, or a puncture was made in a vascular part of the ear with a sharp scalpel. By this procedure a good flow of cutaneous blood was secured, the first few drops being wiped away and the following drops absorbed by the unsized paper used in Tallquist's method of determining the h~moglobin percentage. This was the only method employed in estimating the h~mog lobin, and, while g reat accuracy cannot be claimed (Cabot allows an error of 10 per cent. for it), it has the advantage of simplicity and availability in all cases under all circumstances. Smears were mad e o n specially cleaned slides, three or four being made in each case. The blood for estimating the erythrocytes and leucocytes was drawn in the same pipette, using Toisson's diluting fluid, and in every case securing a dilution of I to 100, the estimation being made by the Thoma-Zeiss h~matocytometer with the Zappert-Ewin g ruling. An objection might be made to the lack of proper preparation of the field for making the incision, but the method adopted was not attended by any bad results, such as the introduction of epithelial cells or fat-drops of sebum in excess to interfere with the leucocy te count, a nd it has the advantage of not introducing the error of increasin g the number of leucocytes by manipulation of the part. In estimating the number of red cells 100 squares were counted, on each of two slides, in most cases, and, when the results were not close, other slides were prepared until what was thought to be a fair average was secured. The whol e ruled space, or 9 to 10· sq. mm., was counted on two or more slides for the white cells. The differential count of leucocytes was macle in all cases from slides stained with Jenner's stain as modified by Dr C. W. Hastin g, which was found to be exceedingly satisfactory. Numerous smears stained by Wright's method and by Ehrlich's tri-acid stain revealed little not brought out by Jenner's. A total of at least 500 leucocytes was counted in each case, and any unusual results were confirmed or corrected by examination of additional slides, even in some cases by securing more films from the subject in question. While Drake apparently recognises only three varieties of leu cocytes, no difficulty is experienced in distinguishing five varieties, unless there be some hesitancy about separating the lymphocytes and large mononuclears. Certainly the mast cells cannot be over-
GENERAL ARTICLES.
21
looked when stained by Jenner's stain. The lymphocytes are slightly larger than the red cells, generally spherical in shape, from 7'4 to 10 microns in diameter, and averaging about S'5 microns. They have a blue nucleus which occupies nearly the whole cell, showing in most cases a crescentic-shaped, darker or lighter blue portion of the cell body at the periphery. Much larger cells, I I to 14 microns in diameter, are found in far fewer numbers, with round nuclei, occupying relatively less of the cell space, while the more typical large mononuclear leucocytes are similar in size and have a mediumsized nucleus, light blue and bean-shaped, and show a large portion of the cell body stained a different shade of blue. The polynuclears closely resem ble those of the horse in size (10 to I4 microns in diameter) and in morphology of the nucleus, which is exceedingly polymorphous, and takes the blue stain; but the cell body is, in most of the specimens, barely visible unless the procedure in staining is very careful. By carefully timing the staining, washing, and rapidly drying in the air, the cytoplasm is seen to be a faintly pinkish colour with a circular outline to the cell body. The eosinophiles are comparable to the polynuclears in size or slightly larger, and have a bi-Iobed nucleus, the two parts of which are connected by a thick band and take the basic stain. The cell body is granular, but the granules are not nearly so large and distinct as those found in the eosinophile of the horse, but are more numerous, estimated at 100 more or less, and have a strong affinity for the eosin stain. The mast cells are about the same size as the preceding and have a similar shaped nucleus, but the cell body possesses granules of a smaller size, more distinct in outline, and of a purple colour. The granules lie both in the cytoplasm surrounding the nucleus and in a position superposed to the nucleus. In hurriedly made smears the erythrocytes are badly crenated, more especially if the smear is thick; otherwise they are circular in outline, and differ in no material respect from those of the horse or man. Many or most preparations show more or less numerous clusters of small bodies, I to 2 microns in diameter, faintly blue in colour, seeming not to take the stain diffusely. These have no particular relation to either the red or white cells as to location and are presumably blood plates. However, Cabot says the blood plates take the eosin stain, while Ewing admits that they take either the acid or basic stain. In all, the blood of twenty-four different pigs has been examined and the results tabulated. All the cases were considered normal, being neither exceedingly poor nor excessively fat and possessed of the characteristic appetite, varying in size, colour, breed, sex, and source, though mostly from the surrounding farms. Intestinal and other parasites are common in swine in the locality of the Institute, but only a post-mortem examination will reveal to what extent th}! cases examined are infested. An accurate knowledge of the effect of these parasitisms on the blood is wanting. Numbers 13 to IS had recently been weaned and were in good condition, and, it will be noticed, show a very close comparison in results of the blood counts. Pig 19 was the only pure white one in the lot and differed materially from the others in conformation, having an undesirable length of nose and legs; yet his lymphocyte count shows a large percentage, far
GENERAL ARTICLES.
22
larger than in many of the much better nourished animals. From this it seems rather difficult to point any constant relation between the percentage of the different varieties of Jeucocytes and the nutrition of the animal, although it would seem that the lymphocytes might increase in fatter animals. In Case 9, the only black and white spotted pig, there is an unaccounted-for increase in the eosinophiles ; consequently, they are not considered in figuring the general averages. The averages of the different varieties of leucocytes are calculated both by working out the percentages from the average number of leucocytes (I9,oOO) and by using the results in the different columns. These results compare favourably. All of the too few standard works on hcematology and blood have been consulted, and only a few references of the more important articles immediately connected with the subject in hand are included in the references. This work has been done under the direction of Dr C. A. Cary, whose constant advice and suggestions have materially assisted in accomplishing satisfactory results. Thanks are due to Dr Cary's assistant, Mr C. T. Butler, B.S., for his intelligent assistance and co-operation, both in securing the blood and in counting the erythrocytes. TABLE
I. -
Ko.
Se.c.
Date 0/ TakttlY.
Age.
II". Pel' {'lilt.
EI·lIt1t"ocl'le.~.
Pel'
Clnllt.
Leucoc,1Ite-<,
-
Reillur/.s.
Pel' emllt.
- - - - - - - - - - - - ------ - - -- - - - - - - - 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.
17-
18. 1H. 20. 21. 22. 23. 24.
Max.
l\1in. Av.
C. m. S. f. S. f. S. f. S. f.
S. f. S. f.
11/13/06 11/Hi/06 11/16/06 11/16/06 11/20/06 n/21/06 11/22/06 11/23/06 11/26/06 11/27/06 11/28/06 11/29/06 12/4/06 12/3/06 12/4/06 12/5/06 12/6/06 12/7/06 12/10/06 12/11/06 12/11/06 12/12/0H 12/12/06 12/13/06
14m. 10 F.
-
M. F.
C.m.
l\1. 111. l\1. 111. 111. 1II. F.
C. m. F.
l\I.
-
-
-1 mo. 90 4 mo. 100 4mo. 85 4 mo. 95 4 mo. 90 4 mo. 00 4 mo. 90 4 mo. 80 5 mo. 80 3 mo. 85 85 3 mo. 95 5 mo. 2'5 mo. 90 2'5 mo. 85 2';' mo. 85 2'5 mo. 85 2',1 mo. 85 2'5 mo. 85 Hmo. HO G mo. 85 6mo. 80 100 6mo. G mo. 85 6 mo. 90
8,200,000 7, 72n, 000 7,750.000 7,180,000 8,700,000 8,050,000 \ 8,800,000
18,000 22,000 18,300 10,000 20,000 14,500 19,000 22,000 1R,000 16,000 21,500 17,000 22,000 12,000 16,000 20,000 21,500 17,000 9.000 25,000 19,000 16,500 23,500 24,000
100 80 88
8,800,000 6,800,000 8,450,000
25,000 9,nOO 19,000
6 mo. 2':; mo. 4 mo.
7,500,000 8,000,000 8,400,000 8,400,000 7,600,000 7,300,000 8,000,000 7,000,000 7,100,000 6,800,000 7,700,000 R,OOO,OOO 7,900,000 8,400,000 8,600,000 8,800,000 8,~80,000
Black, in fair condition.
"
" " " " Bl~ck. " Bl~ck and white,
" " "
under.ized Black and white, spotted. Black, runt rnptured. Small, black. Black, good condition. Black, just weaned.
" "
"
"
" " " " "
Ail white. Black. Black, undersizf'a. Black.
B{~ck, large, well fed.
--
I
Note.-C, castrated; S, spayed; lIf, Male; F, female.
---
23
GENERAL ARTICLES,
TABLE
II,
Differential Count of Lellcoc)'tes, Ly IIIphocytes. No,
Le1!c()cptt!x.
Per
Lar{fe 1lfononucleal',
Polynucl~ar.
-----
cml/~.
I
Mast Cells,
Eo,·"finopltiles.
I cent. 1''''1 1'.0.. 1cent. Pel' I
Pel' I lio. . cent. No. I No. - -- - -- - - -- - - - - - - - - - - - - - - - - -
Pel' cent.
1. 2, 3, 4, 5, Ii. 7. 8.
fl. 1O, 11, 12. 1:~,
14. 15,
]6,
17, IS, 19, 2O, 21. 22, 2:t 24, l\Ia", lIIin, Av, 2nd Av,
I I
No.
Pel' cent.
18,000 22,000 18,300 10,001) 20,000 14,500 19,000 22,000 18,000 16,000 21,500 17,000 22,000 12,000 16,000 20,000 21,500 17,000 9,500 25,000 19,600 16,500 23,500 24,500
30 32 52'6 37 46 45'4 44'4 48'6 43 60 44 47 60 6ii'8 51'4 79'8 57'4 45 68'S 54 67"4
5,400 7,040 9,626 3,700 9,200 6,583 l',436 10,(;92 7,740 9,600 9,460 7,990 13,200 7,896 8,224 15,960 ]2,341 7,650 6,536 13,500 13,210'4 ,Ill 8,085 12,4;)5 53 50'6 12,397
6'4 1,152 8 1,760 1,09H 6 1,000 10 2'8 ' 560 ;; 725. 3'(j Ij84 4·t) 1,012 9'8 1,764 4'2 672 4'8 1 1,033 5'4 , 918 2'0 57:! 1'1) ! 197 4 640 400 2 2 430 2'4 400 152 1'6 4 1,000 '8 1568 2'6 ! 429 ! 70,1 :\ 2'8 686
25,000 9,500 HI, 000
79'S 15,960 30 3,700 51'6 9,400 !l,890
10
-
'8 I 4'6 I --
1,760 1Ii2 715 874
10,800 2'2 12,540 2 6,954 1'2 5,000 no 1'8 45'6 9,120 5 44 6,380 4 44 8,360 6'6 38 8,360 8 3,780 25'S 21 :~,360 21 9'2 34'S 7,480 11 39'2 6,664 6'4 34'2 7,524 3 26'2 3,144 3'S 6,400 40 4 13 2,600 4'S 5'4 ~5 7,525 47'4 S,058 5 2,5 2,37n 4 33 8,250 8 23'(j 4,625'6 7'8 40 6,660 6'6 36'4 8,554 7 38'8 9,50G 6'4 60 12,540 11 1;\ 2,375 1'2 6,820 5'2 37 -7,030 60 57 38
396 440 219'5 180 1,000 580 1,254 1,760 4,644 1,472 2,3/i5 1,088 660 466 640 960 1,161 8.'iO 380 2,000 1,528'8 1,089 1,645 1,568
1'4 1 2'2 1'2 '6 l'Ii 1'4 'S '4 5'6 5'4 2 '2 'S '6 '4 '2 '2 '6 1 '4 1'8 '6 1'4
255 220 402'5 120 120 232 266 176 72 S96 1,162 340 44 96 96 80 43 34 57 250 78'4 297 141 343
2,360 180 1,050 988
5'6 '2 1'3
1,162 34 245 247
-
INDIAN EQUINE PIROPLASMOSIS.
By
CAPT,
A.
J,
WILL1AMS,
F,R,C.V,S" A.V,c.
History,
THIS disease has been known in India for a considerable time under the name of bilious fever, and in South Africa an identical disease was commonly known as biliary fever, According to Hutcheon, it was described by veterinary surgeons in Natal, as far back as 1883, as the biliary form of horse-sickness and as anthrax fever; Hutcheon also states that, although it strongly resembled horse-sickness, he was convinced that it was of a distinct origin and amenable to different treatment, • In the same article Bowhill 5tates that, as far as he was able to ascertain, Hutcheon's conclusions are accepted by all recent writers except Edington, who divides horse-sickness into two forms, one of which he describes as a non-virulent form with a malarial type of fever and in which parasites are found, Theiler admits no co-relation between horse-sickness and equine