NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
December, 1953]
A.
F.
HATHOOT
271
Therefore it could be safely concluded that the younger are the sheep the more was
the red cells count.
Hæmoglobin percentage
The percentage of hæmoglobin varied from 72 to 54 in all groups, the averages
recorded for the four groups were 70.5 for new-born, 72 for lambs, 62.5 for the young
sheep and 54 for adult sheep. Here too, the hæmoglobin percentage tended to be
high in young animals and low in adult ones.
White blood corpuscles
The number of white blood corpuscles per cu. mm. for sheep as a whole varied
from 8,000 to 10,000. Averages being 10,000 for new-born lambs, 8,500 for lambs,
9,200 for young sheep and 8,000 for adult sheep. The lowest average recorded
was that of the adult sheep. The highest average was recorded by new-born lambs,
then it was observed that the averages tended to be lowered in lambs while it increased
a little in young sheep.
Differential count
A. The neutrophile granulocytes
The neutrophile cells, according to their percentages occupy a second place to the
lymphocytes in all groups except animals of group 1 new-born lambs, where the
neutrophile cells recorded a higher percentages than the lymphocytes. The average
percentage of neutrophiles were 64.5, 39.0, 43.0 and 41.0 for the four groups
respectively.
B. The lymphocytes
The lymphocytes occupied as a whole the first place in all groups of animals
except the new-born lambs. This trend was similar to that of the neutrophiles.
Averages of lymphocytes were 44.0, 55.0 48.5, and 50.0. per cent respectively for
the four groups.
C. The eosinophile granulocytes
These cells are not so numerous as the lymphocytes and neutrophiles. The
percentage of these cells in the circulating blood in the four groups varied widely.
The percentage of eosinophiles increased in a parallel direction with the age.
The increase in it was lowest in young animals and highest in adult ones.
The average percentages recorded for the animals studied were 0.2, 2.5, 4.0 and
5.7 respectively.
D. The basophile granulocytes
These cells are very scanty in the blood as they were not demonstrable
in the majority of 60 animals examined.
Averages were 0.1, 0.5, 0.2 and 0.3 for the four groups.
E. The monocytes
These cells were not very numerous in the blood. They showed high counts in
young sheep, being 4.3 per cent and low counts in new-born lambs being, 1.2
per cent.
Close agreement was noticed between our records and that of Burnett [1917]
and Holmann [1945] as shown in Table II.
272
THE STUDY OF NORMAL BLOOD OP SHEEP [Vol. XXIII, Part IV,
TABLE II
Comparison of count of cells by two authors
Author
Burnett [1917]
Holmann [1945]
Neutro¬
philes
37
17
Lympho-
cytes
53
74
Eosino-
philes
1.7
5.0
Basophile
0.3
1.0
Monocytes
8
3
SUMMARY
Sixty normal samples of blood were examined for R. B. C., W. B. C., H.B. per
cent and differential count. The results are as follows:
(1) The number of red blood corpuscles per cubic millimeter of blood varies
with the age of animals. The counts being higher in the new-born
sheep and low in the adult ones. Lambs and young sheep were inter-
mediate. Males recorded high counts than those of females.
(2) The hæmoglobin percentage showed the same trace of R.B.C. It was
higher in young animals than in the older ones.
(3) The white blood corpuscles counts also behave in a similar manner, being
higher in new-born lambs and lower in adult sheep.
(4) The neutrophiles were found to be higher in percentage than the lympho-
cytes in the new-born lambs.
(5) The lymphocytes were of high percentage in all cases except in the new-
born lambs.
(6) The eosinophiles were found to increase in a parallel direction with age,
they were lowest in the young and highest in adult ones.
(7) The basophiles were the lowest cells in the blood.
(8) The monocytes were not very numerous, they were low in the new-born
lambs.
REFERENCES
Burnett, S. H. [1917[. The clinical pathology of the blood of domesticated animals, 44
Bethe, M. [1892]. Sehwalbe-Morpholog. Arbenten, i, S. 207
Cohnstein, J. [1884]. Aroh. f. gesamm. Physiol. 34, S. 233
Giltner, Ward. [1908]. Ann. Vet. Rev. 33, 557
Holmann, H. [1945]. J. Comp. Path. Therap. 55, 146
Storoh, A. [1901]. Inaug. Diss, Born.
Warthin, A. S. [1903]. Contr. Med. Res. dedicated to Vughan, Ann. Arber. 216
Wetzl, J. [1910]. Zeitschr. f. Tiermed, 14, I.
Wiendick, K. [1906]. Arch. f. wisson. u. prakt. Tierkeilk 32, 113
Woltmann, H. [1895]. J. Exp. Med. 7, 119
COMPARISON OF FOUR EJACULATES COLLECTED IN
RAPID SUCCESSION FROM THE INDIAN WATER
BUFFALO (B. BUBALIS)
By S. S. PRABHU and U. D. SHARMA, Division of Animal Genetics, Indian
Veterinary Research Institute, Izatnagar
(Received for publication on 14 March 1953)
C OMPARATIVELY little information on the factors affecting the semen of
the Indian water buffalo, B. bubalis, is available in literature. Kushwaha
and Bhattacharya [1951], Kushwaha, Mukherjee and Bhattacharya [1952], and
Malkani and Patel [unpublished] present data on the seasonal variations in the
quality of buffalo semen. Kushwaha and co-workers had worked on the semen of
buffalo breed commonly found in Uttar Pradesh, while Malkani and Patel had
studied the Surati breed of buffaloes. Prabhu and Bhattacharya [1951], compared
the first and second ejaculates collected in rapid succession from eight buffalo bulls
at weekly intervals. They found that while highly significant difference existed
between bulls, out of the 12 criteria of semen quality studied, only percentages
of abnormal and dead sperm showed statistically significant difference between
ejaculates, while the difference in the total number of spermatozoa in the ejaculate
approached five per cent level of significance. In the present paper four ejaculates
taken in quick succession from 12 buffalo bulls belonging to the breed commonly
found in Uttar Pradesh are analysed and compared.
METHODS
The twelve bulls selected for the study were purchased locally so that their
exact ages were not known. They all possessed full mouths and were healthy and
vigorous. Preliminary trials showed that they were suitable for artificial service.
They were kept on a uniform schedule of rations, which consisted of bhoosa or hay
given ad lib and concentrates made up of two parts crushed oats and one part linseed
cake given at the rate of 3 lb. per 800 lb. live-weight. In addition, each bull was
fed 10 lb. of green grass. The experiment was started on 26 May, 1952 and the last
collections were made on 16 July, 1952. To facilitate work, six bulls at a time
were studied. Collections from the two groups of six bulls were made at an interval
of 24 hours. For the purpose of analysis, however, all collections were treated as
if made on the same day. Each ejaculate was studied for the following criteria of
semen quality.
(i) Volume, (ii) Initial motility, (iii) Sperm concentration, (iv) Total spermatozoa
in an ejaculate and (v) Initial pH.
In addition, reaction time was also recorded.
Altogether, per bull eight collections each consisting of four ejaculates were
taken and studied.
RESULTS AND DISCUSSION
In Table I are presented the averages of the various items of semen criteria and
reaction time for the four ejaculates averaged over bulls. The data of the analys is
of variance [Snedecor, 1947] carried out is presented in Table II.
273
7 AR/53
11
274
COMPARISON OF FOUR EJACULATES OF BUFFALO [Vol. XXIII, Part IV,
TABLE I
Comparative statement of the various semen criteria and reaction times in the four ejaculates
Serial
number
1
2
3
4
5
6
Description of item studied
Volume (c.c.)
Initial motility
Sperm Concentration (m/c.c.)
Total sperm in ejaculate (m)
Initial pH
Reaction time (in seconds)
Order of ejaculates
I
Average 2.14
Range (0.2-9.0)
Average 2.69
Range (0—5.0)
Average 835
Range (60-3140)
Average 1660
Range (12-9225)
Average 6.81
Range (6.6-7.2)
Average 32.69
Range (5-226)
II
2.26
(0.3-5.7)
2.31
(0—15.0)
1109
(100-2670)
2388
(40-6040)
6.83
(6.6-7.2)
80.28
(4-1032)
III
1.18
(0.1-5.0)
1.89
(0—5.0)
787
(17-1830)
946
(22-6950)
6.93
(6.6-7.3)
121.11
(5-2445)
IV
1.30
(0.1-4.1)
1.01
(0 -4.5)
815
(100-1860)
1046
(14-2975)
7.02
(6.6-7.4)
269.31
(5-2505)
Critical
difference
0.260
0.4033
117
299
0.0495
83.20
TABLE II
Analysis of variance
Variation
Bulls
Ejaculates
Bulls × ejaculates
Error
D.F.
11
3
33
336
(i) Volume
M.S.
14.8354
26.3666
3.0633
0.8852
F.
16.75**
29.78**
3.46**
..
(ii) Initial
motility
M.S.
13.87
50.01
1.58
1.99
F.
6.96**
25.13**
..
..
(iii) Sperm Con-
centration
M.S.
2404330
2143095
717023
168269
F.
14.28**
12.73**
4.26**
..
(iv) Total sperms
in ejaculate
M.S.
14916791
42449977
3028888
1095480
F.
13.61**
38.75**
2.76*
..
(v) Initial pH
M.S.
0.13
0.88
0.03
0.03
F.
4.33**
29.33**
1.00
..
(vi) Reaction time
M.S.
685781
1003509
314112
86516
F.
7.92**
11.50**
3.63**
..
*Significant at five per cent level
**Significant at one per cent level
December, 1953] S. S. PRABHU AND U. D. SHARMA 275
(i) Volume
There was a highly significant difference in the total volumes of semen collected
from the different bulls. The minimum was 21.9 c.c. (for eight collections with
four ejaculates per collection) given by Bull No. 16 and the maximum was 90.9 c.c.
given by Bull No. 24 which works to 0.68 c.c. and 2.81 c.c. per ejaculate respectively.
There was also great variation in total volume of semen collected in the different
ejaculates. The respective ranges for total volumes collected in 32 ejaculates per
bull were:
First ejaculate
Second ejaculate
Third ejaculate
Fourth ejaculate
5.4 c.c. to 33.4 c.c.
6.0 c.c. to 35.5 c.c.
2.0 c.c. to 16.7 c.c.
3.8 c.c to 22.6 c.c.
In general, larger semen volumes were obtained in the first two than in the subse-
quent ejaculates. Here again, individual behaviour of a few bulls was different.
Thus Bull No. 37 gave a total semen volume in the fourth ejaculate, which was
larger than the other ejaculate totals. Bulls No. 16, 25 and 38 had fourth ejaculate
totals larger than their respective first and third ejaculate totals. On an average,
the difference among ejaculates was statistically highly significant. The critical
difference for testing significance between ejaculate averages worked out to 0.2690 c.c.
Application of this test showed that while there was no significant difference between
first and second ejaculates, both these ejaculates varied significantly from the
remaining two ejaculates. Besides as the interaction 'bulls × ejaculates' came out
to be significant, it also showed that the difference in the ejaculates had varied
significantly from bull to bull.
(ii) Initial motility
As in the case of the semen volume, initial motility also showed highly signifi-
cant variation from bull to bull. The lowest average initial motility was found in
Bull No. 20 with 1.27 and the highest in Bull No. 16 with 3.00. Among ejaculates,
differences were highly significant. While there was no statistically significant
difference in the average motility of first and second ejaculates, both these ejaculates
differed significantly from the remaining two ejaculates. This shows that on an
average the third and fourth ejaculates contain semen of poorer motility-rating than
in the first two ejaculates.
(iii) Sperm concentration
Among the bulls, the lowest average sperm concentration namely 328 millions/c.c.
was found in Bull No. 20, and the highest namely 1376 millions/c.c. in Bull No. 16.
Bull to bull variation was highly significant, so also was the difference among
ejaculates. There was no significant difference among the first, third and fourth
ejaculates, while all of them differed significantly from the second ejaculate which
contained the highest average concentration of sperm. The interaction 'bull X
ejaculate' came out to be highly significant, showing that the sperm concentration
in the four ejaculates varied significantly from bull to bull.
7 AR/53
12
276
COMPARISON OF FOUR EJACULATES OF BUFFALO [Vol. XXIII, Part IV,
(iv) Total sperm in ejaculate
The lowest average total sperm in an ejaculate was found in Bull No. 20, and
the highest in Bull No. 24. The respective figures were 553 millions and 3045
millions. Bull to bull variation in total sperm in ejaculate was highly signifi-
cant. So also was the difference among the ejaculates. The second ejaculate
differed significantly from the remaining three ejaculates; while among the remain-
ing three ejaculates, the first differed significantly from the third and fourth. These
results also show that on an average the first two ejaculates are better than the
subsequent two ejaculates. In addition, the difference among ejaculates had
varied significantly with bulls, which was shown by the highly significant 'bull x
ejaculate' interaction.
(v) Initial pH
As in previous criteria of semen quality, pH showed highly significant varia-
tion in case of both bulls and ejaculates. It was noticed that with the frequency
of collection the pH turned from acidity to definite alkalinity. There was no signi-
ficant difference in the pH between first and second ejaculates, while both these
ejaculates differed significantly from the subsequently drawn two ejaculates. These
results also suggest that the first two ejaculates on an average are better than the
subsequent two ejaculates.
(vi) Reaction time
Reaction time showed highly significant difference from bull to bull and from
ejaculate to ejaculate. Besides, the difference among ejaculates also varied signi-
ficantly from bull to bull. On an average, the first two ejaculates individually
differed significantly from the remaining two ejaculates, while the difference between
the first and second was not statistically significant.
The results presented in this paper confirm the findings of Prabhu and Bhatta-
charya [1951], as far as the first two ejaculates taken in rapid succession are con-
cerned. Further, in all the five items of semen criteria and reaction time compared,
the first two ejaculates appear to be superior to the subsequent two ejaculates.
It was observed that the third and fourth ejaculates contained less semen volumes,
of lower initial motility rating, of lower sperm concentration, of lower total number
of sperm and of higher pH value than the first two ejaculates. Besides, the reaction
times in the former cases were higher. It appears therefore that it is undesirable
to take more than two ejaculates at a time from the buffalo bulls when a schedule
of collections at weekly intervals is followed at the artificial insemination centres.
Even so, there could be exceptions as seen by the large bull to bull variation and
significant 'bull X ejaculate' interaction. A safer course may be to follow generally
the suggestion made in this paper and introduce modification only under special
circumstances.
SUMMARY
1. Four ejaculates taken in rapid succession at weekly intervals from twelve
buffalo bulls of the type commonly found in Uttar Pradesh were analysed in respect
of five semen criteria and reaction time.
December, 1953] S. S. PRABHU AND U. D. SHARMA 277
2. On the basis of the results obtained, it is suggested that, it is undesirable to
take more than two ejaculates at a time from buffalo bulls, though in special cir-
cumstances depending upon the individual behaviour of the bulls a slight departure
from this rule may be made.
ACKNOWLEDGEMENT
The authors are indebted to Dr P. Bhattacharya, Head of the Division of Animal
Genetics, Indian Veterinary Research Institute, Izatnagar, for his keen interest
and encouragement in their work. They are grateful to Dr S. Dutta, Director,
Indian Veterinary Research Institute, Izatnagar, for providing all facilities. Their
thanks are also due to Shri V. N. Amble, Statistician (A.H.) of the Indian Council
of Agricultural Research for advice on the statistical treatment of the data.
REFERENCES
Kushwaha, N. S. and Bhattacharya, P. (1951). Seasonal variation in reaction time and semen
of buffalo bulls. Proc. 38th Indian Sci. Congr. Bangalore, 3, 248
Kushwaha, N. S., Mukherjee, D. P. and Bhattacharya, P. (1952). Seasonal variation in reaction
time and semen of buffalo bulls. (Unpublished)
Malkani, Mohina, C. and Patel, M. D. (1952). Seasonal variations in the quality and quantity of
semen of buffalo bull. (Unpublished)
Prabhu, S. S. and Bhattacharya, P. (1951). Comparison of first and second ejaculates collected
in rapid succession from the Indian water buffalo (B. bubalis) Indian J. vet. Sci. 21(4),
257-262
Snedecor, George, W. (1947). Statistical Methods. The Collegiate Press; Inc., Iowa
FERTILITY OF BULL SEMEN PRESERVED FOR LONG
PERIODS
By S. S. PRABHU, K. D. BHAYA and P. BHATTACHARYA, Animal Genetics Division,
Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh
(Received for publication on 31 March 1953)
W ITH the rapid development of air transport, the time factor involved in long
range inseminations has been considerably shortened, making possible the
widest use of available semen from high quality pedigree sires on a global scale.
As compared to Milovanov's statement [1932] that the distance to which bull semen
is to be transported should not exceed two hours' journey and that not more than
six hours should lapse between collection and insemination, Lambert and MacKenzie
[1940] reported successful impregnation with semen air lifted from Beltsville,
Maryland to Piran in the Province of Buenos Aires, with the total time-lapse of
seven days between collection and insemination. Cows in England have been
successfully inseminated with semen sent by air from Denmark [Edwards, Walton
and Siebenga, 1938] and those in Italy from semen air lifted from South and North
America [Bonadonna, 1949]. In India, successful inseminations have been per-
formed with Hariana semen sent either way between Izatnagar and Calcutta. In
addition, over 250 inseminations have been performed at Cochin, Ernakulam and
Colombo from air-borne semen from Sindhi bulls stationed at the Animal Genetics
Division's field station at Indian Dairy Research Institute, Bangalore. In India,
among the various problems associated with long range paternity, the economics of
the time factor involved between despatch of semen from the site of collection to
the sites of insemination is an important one. This is specially so in cases where
the animals to be inseminated are located right in the interior, and are reached only
through primitive modes of conveyance over difficult road and terrain. In view of
the heavy expenditure inevitable in such cases, one has to see whether it is repaid
by a proportionately reasonable level of fertility. To investigate this problem, the
present study was undertaken.
METHODS AND RESULTS
In order to ensure that sufficient number of cows would be available on a given
day for insemination, the corpus lutea of cows in groups of 6-10 from the experimental
herd at Izatnagar were manually removed. The animals usually came on heat
from 3rd to 4th day after the treatment. In this way oestrus cycles of a group of
cows were so adjusted as to ensure sufficient number of cows on heat to be ready at
about the dates required. Semen samples of good quality from two young bulls
were in the meantime processed and kept ready so that when the cows came on
heat and/or repeated, semen samples with storage times ranging from 120-241
hours were available for artificial service. In this way, 21 cows were inseminated
279
7 AR/53.
13
280
FERTILITY OF BULL SEMEN
[Vol. XXIII, Part IV,
using semen from Bull No. 1, and 18 cows from semen of Bull No. 2. The results
are shown in Table I.
TABLE I
Details of semen used and the percentage of conceptions obtained
Bull number
1
2
Total
2
Number of
cows used
21
18
39
Number of
inseminations
36
20
56
Number
pregnant
7
2
9
Percentage of
conception
19.44
10.00
16.07
Details of the actual age of semen used in case of conceiving cows is given in
Table II.
TABLE II
Age of semen used in the successful cases
Cow number
1
2
3
4
6
6
7
8
9
Age of semen in hours
216
216
120
241
241
197
168
192
126
Bull number
2
2
1
1
1
1
1
1
1
DISCUSSION
In our study, more than 50 per cent of the successes were obtained with semen
stored for 9-10 days. Such samples according to the unpublished data collected
by Bhatnagar contain on an average 15-20 per cent motile sperm. Of this, a small
proportion differing with semen samples are progressively motile. The actual
percentage of total and progressively motile sperm present in the stored semen
samples depended on the bulls and varied from ejaculate to ejaculate within a bull.
Report of Margolin, Bartlett and Lepard [1943] shows significant correlation between
December, 1953] S. S. PRABHU, K. D. BHAYA AND P. BHATTACHARYA 281
longevity of preserved semen stored at 4ºC. and fertility. In our study, estimations
of motile sperm in semen samples were not carried out. But considering the average
values, an estimate can be arrived at regarding the probable number of motile
sperm present on an average in such samples, and it approximates to the estimate
given by Salisbury and Bratton [1948] as to the number of live sperm that should
be introduced in the reproductive tract of the cows for ensuring conception.
The average over-all fertility for all inseminations works at 16.07 per cent.
If the 16 cows, on which 30 inseminations were performed, are omitted (in view of
their proving to be shy breeders) the overall fertility rate rises to 34.60 per cent.
Further, a certain correction has to be applied for inseminations made at induced
heats, which according to foreign workers, produce lower conceptions. Taking
into account all relevant factors, it may not be considered unreasonable to estimate
that given good quality semen to start with, preservation for 9-10 days at 4ºC.
gives a fertility rate lying between 35-40 per cent. No doubt, this fertility rate is
20 to 25 per cent less than what is normally obtained, yet, in our opinion, the results
offer a reasonably sound basis for undertaking long range paternity programmes,
where the time-lag between collection and insemination is of the order of 9-10 days.
It is to be remembered that the findings reported in this paper pertain to currently
used methods of preservation. With the new technique of preserving sperm at
very low temperatures [Polge and Rowson, 1952] developed in Britain, far reaching
changes affecting long range paternity are expected.
SUMMARY
From a study of 56 inseminations performed with good quality bull semen
preserved upto 10 days at 4ºC., it has been estimated that about 35-40 per cent of
the cows may be expected to hold, which provide a reasonably sound basis for
undertaking long range paternity service, when required.
ACKNOWLEDGEMENT
The authors are indebted to the Director, Indian Veterinary Research Institute,
for providing all facilities for undertaking this work.
REFERENCES
Bhatnagar, D. S. (1952). Unpublished data
Bonadonna, T. (1949). XII Inter. Dairy Congr. 5, 195-198
Edwards, J., Walton, A. and Siebenga, J. (1938). J. Agric. Sci. 28, 503
Lambert, W. U. and Fred McKenzie (1940). Cir. U.S.D.A. 567
Margolin, S., Bartlett, J. W. and Lepard, O. L. (1943). J. Dairy Sci. 26, 983-985
Polge, C. and Rowson, L. E. A. (1952). Proc. II Inter. Congr. Physiol. Path. Ani. Repr,
A. I. 90-98
Salisbury, G. W. and Bratton, R. W. (1948). J. Dairy Sci. 31(9), 817-822