NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
CYTOLOGICAL, PHYSIOLOGICAL AND CHEMICAL
STUDIES OF EGYPTIAN BUFFALO BLOOD
By E.S.E. HAFEZ 1 and A. ANWAR 2
(Received for publication on June 8, 1955)
(with one Text-Figure)
T HE comparative study of the normal values of blood constituents and the
physiological variations which occur in the absence of any definite pathological
condition, is of great academic and practical importance for clinical hematology
and animal husbandry especially in the tropics, since certain blood components
are related to heat tolerance, environmental stress and disease immunity.
Normal hematological values vary considerably amongst different classes of
farm animals. In general the erythrocyte count is inversely proportional to the
body size of the species. In man, horse, swine, dog and cat the neutrophils are
the most frequent leucocytes while in ruminants, laboratory rodents and chicken
the lymphocytes are predominant [Olsen, 1937].
In this respect the hematological values of the Egyptian buffalo Bos, (Bubalus),
bubalis, L. were determined.
MATERIAL AND METHODS
Experimental animals
Thirty healthy adult Egyptian buffalo-cows aged from three to ten years were
available at the Animal Production Research Farm, Faculty of Agriculture, Uni-
versity of Cairo, Giza. Blood samples were taken for the cytological, physiological
and chemical studies during the period from January to April, 1953.
Technique
1. Cytological studies. Four smears were prepared from each blood sample.
They were stained with Delafield Hematoxylin and Eosin, Giemsa stain, Wright's
stain and Leishman's stain. The corpuscle diameters were measured using the
stage micrometer and the micrometric eye-piece. A Thoma-Zeis hemocytometer
was used for the erythrocyte and leucocyte counts.
2. Physiological studies. The specific gravity of the whole blood was measured
using the copper sulphate method of Weises [1950], the pH number using
" Beckman " pH meter, the sedimentation rate after one hour in " Westergeren "
pipettes and the clotting time using " Sabrazs " capillary tubes. Hemoglobin
content was determined colorimetrically using a " Halden " hemoglobinometer
and the hematocrit using " Wintrobe " tubes in a centrifuge of 3000 rotations per
minute for 30 minutes.
(1) Faculty of Agriculture, University of Cairo, Egypt. Present address : The Worcester
Foundation for Experimental Biology, Shrewsbury, Mass., U.S.A.
(2) Ministry of Agriculture, Cairo, Egypt.
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126
STUDIES
OF
BUFFALO
BLOOD
[ Vol. XXVI, Part III,
3. Chemical studies. Whole blood sugar was determined using the method
of Folin and Wu, [1920], the plasma proteins and the non-protein-nitrogen fractions
using the technique of Van Slyke and Cullen [1914], plasma sodium using the uranyl-
zinc-acetate method of Weinbach [1935], potassium using the technique of Looney
and Dyer [1942], calcium using that of Roe and Kahn [1929] and phosphorus using
the technique of Fiske and Subbaraw [1925].
RESULTS
The average diameter and the differential count of the blood cells from normal
healthy animals are shown in Table I (see also Fig. I).
TABLE I
Cytological constants of the blood
Blood cell
Erythrocytes
lymphocytes
Leucocytes monocytes
neutrophils
eosinophils
basophils
Average diameter
(microns)
5.5
6.8
15.7
11.3
11.8
11.8
Count/mm. cu.
Range
5.75 × 10 6 — 7.5 × 10 6
5.6 × 10 3 —9.0 × 10 3
Average ±S.D.
6.8 × 10 6 ±0.5l7
6.7 × 10 3 ±0.679
The specific gravity of the whole blood ranged from 1.050 to 1.070 with an
average of 1.058 gm./ml. The pH number varied within narrow limits of 7.35
to 7.60. The sedimentation rate after one-hour period ranged from 2 to 8 with an
average of 6 mm. while the clotting time ranged from 6.0 to 7.25 minutes with an
average of 6.75 minutes. Hemoglobin content ranged from 11.04 to 15.18 gm.
per 100 ml. of blood with an average of 12.98 gm. The hematocrit value ranged
from 38 to 52 per cent with an average of 44.3 per cent (Table II).
TABLE II
Physiological constants of the blood
Items
Specific gravity
pH
Sedimentation-rate
Clotting time
Hemoglobin
Hematocrit
Unit
gm./ml.
mm./1 hr.
min.
gm./100 ml.
percentage
Range
1.05-1.07
7.35-7.60
2.00-8.00
6.00-7.25
11.04-15.18
38-52
Mean±S.D.
1.058 ±0.012
7.45±0.038
6.00±1.65
6.75±0.065
12.98±1.236
44.3±3.028
C.V.
per cent
1
0.5
29
1
10
7
September, 1956 ]
E.
S.
E.
HAFEZ
AND
A.
ANWAR
127
FIG, 1 Frequency distribution of blood counts in 30 healthy buffalo cows. Black columns
indicate erythrocytes ×10—6 and white columns indicate leucocytes 10—3
128
STUDIES
OF
BUFFALO
BLOOD
[Vol. XXVI, Part III,
Blood sugar per 100 ml. ranged from 72 to 90 mg. with an average of 81.4 mg.,
while plasma proteins ranged from 6.82 to 7.68 with an average of 7.43 gm./100 ml.
plasma. The total non-protein nitrogen per 100 ml. blood ranged from 25 to 42 mg.
The average amounts of sodium, potassium, calcium and phosphorus per 100 ml.
plasma were 415, 26, 10 and 28 mg. respectively (Table III).
TABLE III
Chemical constants of the blood
Items
Sugar
Plasma proteins
Plasma
non-protein
nitrogen.
Plasma sodium
Plasma Potassium
Plasma calcium
Plasma phosphorus
Unit
mgm./100 ml.
gm./100 ml.
mgm./100 ml.
mgm./100 ml.
mgm./100 ml.
mgm./100 ml.
mgm./100 ml.
Range
72-90
6.82-7.62
25-42
400-430
23-29
9-11
26-31
Mean±S.D.
81.4±3.411
7.43±0.266
36±3.142
415±6.53
26±1.00
10±0.156
28±1.032
C.V.
per cent
4
4
8
2
3
2
3
DISCUSSION
The blood picture of the animal is a result of an interaction between environ-
mental and hereditary and nutritional factors. The blood corpuscles of the buffalo
possess the same staining affinity as that of other mammals. The neutrophils,
eosinophils and basophils are similar to those of cattle in shape and granulation.
Neutrophillic granules are spherules, different from those of chicken where they are
large and elongated and from those of guinea pig and rabbit where they are rod-
shaped. Eosinophillic granules are round-shaped and larger than those of neutro-
phils but still smaller than the specific eosinophillic granules of the horse. The
basophillic granules are as large as the granules of eosinophils and evenly distributed
in the cytoplasm. In certain species (e.g. the dog) they are assembled in a small
compact group. The erythrocyte diameter is almost the same as that of the horse,
September, 1956 ] E. S. E. HAFEZ AND A. ANWAR 129
ox, and larger than that of sheep. Hematological values of domestic animals have
been reviewed by Dukes [1952], Maximow and Bloom [1952] and Trautmann and
Fiebiger [1947].
In the buffalo, the erythrocyte count is higher than in cattle and lower than in
sheep and horse, while the leucocyte count is less than in cattle, horse and sheep.
The study of the differential count shows that the percentage of lymphocytes,
monocytes and eosinophils are higher in the buffalo than in sheep and horse and
lower than in cattle. The neutrophils are lower than in sheep and horse but higher
than in cattle. The basophils are almost of the same frequency (1 per cent) in all
farm animals.
The specific gravity of the buffalo blood is slightly lower than that of the horse
and higher than that of cattle and sheep. The pH number varies within narrow
limits, it is within the normal range of pH of mammalian blood. The sedimentation
rate in the buffalo is much higher than in cattle and lower than in horse, while the
clotting time of buffalo blood is longer than in sheep, shorter than in horse and
similar to that of cattle. A comparison between the hematological values of the
Egyptian and Indian buffalo is shown in Table IV.
TABLE IV
Hematological values of Egyptian buffalo as compared with that of the Indian
buffalo
Item
Red count
Hemoglobin
Hematocrit
Plasma proteins
Sugars
Author
Unit
million/mmc.
gm./100 ml.
Percentage
gm./100ml. plasma
mg./100 ml.
Egyptian buffalo
6.8
12.96
44.3
7.43
81.4
Present experiment
Indian buffalo
6.1
7.7
35.5
7.46
79.4
Kehar and Murty
[1951]
The hemoglobin content as well as the sugar level is higher in the buffalo than
in all other farm animals. The plasma proteins and the non-protein-nitrogen are
similar to that of cattle and higher than that of other farm animals. As far as the
minerals are concerned, the buffalo blood is characterized by a relatively high con-
tent of phosphorus and a low content of calcium and sodium. Potassium is almost
the same in all farm animals.
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STUDIES OF BUFFALO BLOOD
[ Vol. XXVI, Part III
SUMMARY
Thirty healthy adult Egyptian buffalo-cows aged from three to ten years were
available at the Animal Production Research Farm, Faculty of Agriculture, Uni-
versity of Cairo, Giza, Egypt. Cytological, physiological and chemical constants
of their blood were studied.
1. In the buffalo, the blood corpuscles possessed the same staining affinity as
in other mammals. The erythrocyte count was 6.8 × 10 6 /mm.c. while the leucocyte
count was 6.7 × 10/mm.c. In the leucocytic differential count the percentage
of lymphocytes, neutrophils, monocytes, eosinophils and basophils were 51, 36,
8, 5 and<1 respectively.
2. The specific gravity of the buffalo blood was 1.058 gm./ml. The pH number
was 7.45 the sedimentation rate after one hour was 6 mm. and the clotting time
was 6.75 minutes. The hemoglobin content was 12.96 gm./100 ml. blood and the
hematocrit was 44.3 per cent.
3. In buffalo the blood sugar was 81.4 mg./100 ml. while the non-protein-
nitrogen ranged from 25—42 mg./100 ml. blood. The plasma proteins were 7.45
gm./100 ml. plasma while the sodium, potassium, calcium and phosphorus/100 ml.
plasma were 415, 26, 10, and 28 respectively.
4. The comparative study of certain cytological, chemical and physiological
features may be of academic significance to investigate the evolutionary trends of
species and breeds.
ACKNOWLEDGMENT
The authors are thankful to Prof. A. L. Badreldin, Animal Production Depart-
ment, Faculty of Agriculture, University of Cairo and Dr M .H. Maghrabi of the
Animal Breeding Section, Ministry of Agriculture, Egypt, for providing facilities and
help.
REFERENCES
Dukes, H. H. (1952). The Physiology of Domestic Animals. 6th Ed. Comstock, Ithac, New York
Fiske, C. H. and Subbaraw, Y. (1925). J. Biol. Chem. 66, 375
Folin, O. and Wu, H. (1920). J. Biol. Chem. 41, 367
Kehar, N. D. and Murty, V. N. (1951). Indian J. vet. Sci. 21, 13
Looney, J. M. and Dyer, C. G. (1942). J. Lab. Clin. Med. 28, 355
Maximow, A. A. and Bloom, W. (1952). A Textbook of Histology. 6th Ed. Saunders Co., Philadel-
phia
Olsen, C. (1937). Cornell Vet. 27, 235
Eoe, J. H. and Kahn, B. S. (1929). J. Biol. Chem. 81, 1
Trautmann, A. and Fiebiger, J. (1947). Fundamentals of the Histology of Domestic Aniamals. Comstock,
Cornell
Van Sluke, D. D. and Cullen (1914). J. Biol. Chem. 11, 211
Weinbach, A. P. (1935). J. Biol. Chem. 110, 95
Weises, F. (1950). Ital. Sez. I, 22, 101.
ISOLATION OF SALMONELLA-LITCHFIELD FROM AN OUT-
BREAK IN CHICKS IN THE INDIAN UNION
By S. B. V. RAO, Indian Veterinary Research Institute, Izatnagar
(Received for publication on January 3, 1956)
"IN spite of the frequency of paratyphoid fevers in man and the general importance
of the Salmonella problem in public health, including meat and milk inspection,
the animal Salmonella problem has been scarcely touched, the chief reason being
the dearth of trained workers and the mass of general diseases that confronts the
animal pathologist." The authors of this statement, Shirlaw et al., [1945] listed a
few Salmonellae isolated in animals in India by previous workers, viz. Sal. abortus
equi in horses, Sal. gallinarum in chickens, Sal. enteritidis in pigeons, Sal. dublin
in calves and Sal. typhi-murium in hounds. Since the publication of this report,
little has been added to our knowledge of the incidence of salmonella infections in
animals in India probably due to the same reasons. Of late, due to the establishment
of large government poultry farms as a means for increasing food production, the
attention of animal pathologists has been drawn to this problem, mainly to prevent
poultry mortality ; their efforts, however, have been confined to organized farms.
No attempts are made, however, to determine the incidence of salmonellae in the
rural poultry population which is the main source of supply to the markets. The
adult chickens in the villages are most likely to be carriers and reservoirs of these
organisms, particularly due to their feeding habits which are essentially scavenging
in manure dumps, cesspools, and waste lands around hutments. The single factor
which keeps down the spread of these pathogens to man is the cooking of poultry-
meat and eggs before consumption. The handling of these products, the use parti-
cularly by invalids, of eggs in the raw state and in bakery products, and the frequent
presence of chickens within the homes of the villagers, are the public health risks
which cannot be over-emphasized.
Salmonella typhi-murium was first isolated from a mouse epizootic in 1890 by
Loeffler. It has since been reported in many animals and man. Kauffmann [1941]
states that from surveys carried out in England, Germany and Denmark, Sal. typhi-
murium was responsible for an average of 65 per cent of the cases of salmonellosis
in man. Iyer and Rao [1950] reported an outbreak of the disease in India, in a
chicken farm due to this organism. Sal. anatum in ducks was recorded by Rettger
and Scoville [1920], and since then it is known to have caused the disease in poultry
and man in many parts of the world. It was often the cause of serious mortality
among ducks in the Army Development Farms in India in world war II [Rao,
*Resident Fellow Colombo-plan Training National Salmonella and Shigella Typing and Reference
Centre, Laboratory of Hygiene, Ottawa—Department of National Health and Welfare, Canada.
131
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132
ISOLATION OF SALMONELLA-LITCHFIELD [ Vol. XXVI, Part III,
1946]. Dixit [1951-1952] isolated Sal. anatum in baby chicks in Bombay
Province. Pande et al., [1933] isolated a motile strain of Salmonella from baby
chicks, which was subsequently typed as Salmonella bovis morbificans in England.
The author isolated three strains of Salmonella (other than S. pullorum) in
1952-53 from serious outbreaks, two in chickens and one in a rabbitry. All three
were identified by preliminary biochemical and serological tests as Sal. typhi-murium,
and were confirmed by Dr J. D. Blacksland of the Weybridge Laboratory (Annual
Reports I.V.R.I. 52-53). A fourth one was subsequently isolated from baby chicks,
and was taken to Canada for identification and forms the subject of this report.
EXPERIMENTAL PROCEDURE
The organism was isolated from a large military poultry farm where it accounted
for 50 per cent mortality in a lot of 3,000 hatchery chicks, ages ranging from 7 days
to 4 weeks. The general symptoms observed were crowding of the chicks together
under the hover, droopiness and diarrhoea. Over 100 autopsies conducted revealed
no definite symptoms, except pallor of the internal organs and the skeletal muscles.
Rapid whole-blood agglutination test of the laying flock with an "O" antigen, pre-
pared from the specific organism isolated, failed to detect any carriers. The disease
was, however, brought under control by dosing with sulphamezathine (a 16 per cent
solution of the sodium salt) in drinking water, at a level 0.2 per cent sulphamezathine,
for 6 days.
Cultures were regularly obtained from heart-blood and liver on agar slants,
while the organism was isolated from the intestinal contents after enrichment in
brilliant green and tetrathionate broth and subsequent sowing on MacConkey
plates. The strain was maintained for a year on plain agar before final identification
was carried out.
The organism was a Gram-negative, motile rod which possessed the morphological
and cultural characters of the Enterobacteriaceae. Arabinose, dextrose, dulcitol,
galactose, levulose, maltose, mannitol, mannose, rhamnose, soribitol, lactose and
zylose were fermented promptly with the production of acid and gas. Acid and
gas were produced in cellobiose after 7 days, and acid only was found in dextrin
and glycerol in 7 and 14 days, respectively. Adonitol, inositol, inulin, lactose,
raffinose, salicin and sucrose were not fermented. Indole was not formed and the
cultures failed to utilize citrate, liquefy gelatin or decompose urea. The organism
fermented d-tartrate, was Methyl Red positive and Voges-Proskauer negative.
Slide agglutination tests with Salmonella O sera showed the following results:
Agglutination
Pool 1 (O factors 1,2,3,4,5,9,10,12,15,19,21,26,27)
2 (O factors 6,7,8,11,13,14,20,22,24)
3 (O factors 16,17,18,28,29,30)
4 (O factors 35,38,39,40,41,42)
With the sera of Pool 2 : factors 6,7
7
6,8
8
Others
—
++++ —
—
++++
—
++++
++++ —
Delete 'Lactose' on page 132, para. 5, line 3.
September, 1956 ]
S.
B.
V.
RAO
133
The culture appeared to be predominantly in the second phase, reacting strong
ly in "1,2" and single factor "2" H antisera. Phase suppression was carried out
with "1,2" absorbed H sera on swarm agar plates. Three transfers on suppression
plates were necessary before the "1,2" phase could be suppressed and the phase 1
antigenic structure demonstrated. When tested with phase 1 antisera, strong
agglutination was obtained only in "l,v", "l,w" and "v" antisera ; no agglutination
was observed with absorbed "w" antiserum. In tube tests, the organism agglutina-
ted to titre in "l,v" and "v" sera, thus confirming the results of the slide tests.
'Therefore, its antigenic structure is 6,8; l, v-1,2, identified as Sal. litchfield.
DISCUSSION
Sal. litchfield was first isolated by Pomeroy from a turkey poult in 1940 [Edwards
and Bruner 1940] and later from human sources, but has not been previously reported
in chickens. The occurrence of this pathogen in chickens further confirms the present-
ly accepted view of several workers, including that of Seligmann et. al., [1943],
that Salmonellae are not host-specific and that every one of the three hundred odd
ones is potentially capable of producing groups of clinical processes, like gastro-
enteritis, enteric fever, meningitis, etc., and that animals and birds may act as reser-
voirs. It is generally conceded today that animals, particularly poultry, are the
principal reservoirs of most of the Salmonellae, thus emphasizing the great need for
controlling the disease in animals, as a most important public health measure.
SUMMARY
A Salmonella organism isolated from baby chicks, which caused a mortality
of 50 per cent in a large military poultry farm in India, has been identified as Sal.
litchfield (6,8 ; l, v-1, 2). Experimentally, the organism was found to be pathogenic
for young chicks, while the adults became carriers. Sulphamezathine in drinking
water controlled the disease effectively. The need for organized efforts to control
animal reservoirs of Salmonella as a public health measure is stressed.
ACKNOWLEDGMENT
The author expresses his indebtedness to the Government of Canada
for sponsoring his training under the Colombo Plan. He acknowledges with thanks
the advice and expert guidance he received in the technique of the preparation of
the various sera required in general for typing of Salmonella from Dr E. T. Bynoe,
Chief, Bacteriological Laboratory Services, and Dr W. R. Bailey, Bacteriologist-
in-Charge of the National Salmonella and Shigella Typing and Reference Centre,
Laboratory of Hygiene, Department of National Health and Welfare, Canada.
Thanks are also due to Miss R. Laidley for her help.
134
ISOLATION OF SALMONELLA-LITCHFIELD [ Vol. XXVI, Part III,
REFERENCES
Dixit, S. D. (1951). Indian vet. J., 22, 281-283,
—(1952). Indian vet. J., 28 : 425-429
Edwards, P. R. and Bruner, D. W. (1940). J. Inf. Dis 66 : 218-220
I.V.R.I. (1952 and 53). Annual Reports
Iyer, S. G. and Rao, S. B. V. (1950). Indian Poult. Gaz. 34 : 12-14
Kauffmann, F. (1941). Die Bakteriologie der Salmonella Grouppe. Pun. Ejnar, Munksgaard
Copenhagen
—(1951). Enterobacteriaceae. Pun. Ejnar, Munksgaard Copenhagen
Pande, P. G. and Neelakantan, P. R. (1953). Proc. Indian Sci. Congr. 343
Rao, S. B. V. (1946). Indian vet. J. 22 : 279-281
Rettger, L. F. and Scoville, W. (1920). J. Inf. Dis 26 : 217
Seligmann, E., Saphra, I. and Wassermann, H. (1943). Amer J. Hyg., 38 : 226-249
Shirlaw, J. F., Stuart Macdonald, and Hayes, W. (1945). Indian J. med. Res. 33 : 1-3
THE EFFECT OF AUREOMYCIN AND VITAMIN B 12 ON
THE GROWTH RATE OF CHICKS
ALEX REED 1 , and JOHN VELU 2 , Allahabad Agricultural Institute, Allahabad, U. P.
(Received for publication on February 17, 1956)
M ANY investigators have reported increased growth rate and feed efficiency when
chicks are fed rations fortified with aureomycin and vitamin B 12 [Biely et al.,
1952, Machlin et. al., 1952 and Sizemore et. al., 1953]. Groschke and Evans [1950]
reported that aureomycin exerted a definite stimulating effect upon growth when fed
to chicks. Heuser and Norris [1952] compared a number of antibiotics when fed to
chicks and reported variability in response to the same antibiotics in different experi-
ments. They obtained the greatest response with chicks of four weeks of age.
This is in agreement with Scott and Glista. [1950] but at variance with Berg et. al.,
[1950]. Davis and Briggs [1951] in studying the effects of aureomycin, penicillin,
bacitracin and terramycin report that growth stimulation resulted in most cases
and that antibiotics improved feed efficiency. Saxena et. al., [1952] fed penicillin
at three levels of protein, 21, 18, 15 per cent respectively. They reported significant
and consistent response in growth to penicillin at all levels of protein tested. Biely
and March [1951] report no response to antibiotics in rations containing above normal
amounts of vitamins but in rations low in riboflavin, folic acid and or nicotinic acid,
antibiotics gave significant growth increases. Scott et. al., [1952] in studying the
influence of aureomycin on the protein requirements of chicks found the improvement
in protein efficiency no greater than the improvement in feed efficiency, so there was
no protein sparing action from the antibiotic. They also reported that chicks on
a 20 per cent protein ration gave no response to aureomycin. Bose et. al., [1955]
fed Fortracin-6 (bacitracin and vitamin B 12 ) to chicks of eight weeks of age and
obtained increased growth from the antibiotic.
Since most of the work reported on the use of antibiotics for chicks has been done
outside India and with more complete diets than are generally available here, it
was considered desirable to investigate the effects of aureomycin and vitamin B 12
when fed to chicks on a common ration.
EXPERIMENTAL
White Leghorn chicks obtained from Government Farm, Mathura were used in
this study. Three-day-old chicks were randomly distributed into eight groups of
eleven chicks each and randomly assigned to pens in wire cages. The groups were
1 University of Illinois, College of Agriculture, doing teaching and research in Animal Nutrition
at Agricultural Institute.
2 Member staff, Animal Husbandry Department, Agricultural Institute, Allahabad, U.P.
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