NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
276 STUDIES ON PASTEURELLA SEPTICA [Vol. XXVI, Part IV,
Only one strain, i.e. the bile passaged strain 'P 52' was found to lose its virulence
after it had undergone as many as 384 passages but at the same time it showed
evidence of loss in its antigenicity.
(2) Passage of a strain of Past. septica through albino rats
On lines similar to those used for the attenuation of viruses, a virulent strain
of Past. septica was passaged in albino rats, which are normally highly resistant to
this organism, in the hope that the resulting attenuated strain might prove useful
in the successful vaccination of cattle against haemorrhagic septicaemia. Thirty-
one serial passages were carried out by the transference of blood from one rat to
another and cultures obtained at the 5th, 10th, 15th, 20th and 31st passage were
tested for virulence in white rats, guinea-pigs and white mice to see if there was any
evidence of attenuation. The strain was found to have assumed an enhanced viru-
lence for rats and guinea-pigs, but no evidence of attenuation for white mice could
be noticed. In view of this finding, the strain was not tested in cattle, as no appre-
ciable change in its pathogenicity for bovines could be expected.
In view of the above findings coupled with the immunological studies, it was
decided to prepare H. S. Vaccine on the following lines:
(i) To incorporate stock strain P52 of Past. septica after freshly passaging it
through a healthy bovine so as to ensure the full presence of capsular
substance,
(ii) To sterilise the vaccine by the addition of 0.25 per cent formalin and
(iii) to add adjuvants with a view to induce maximal antibody response sus-
tained over a long period.
(d) Having briefly outlined the procedure for the production of vaccines, ex-
periments were undertaken to select a suitable medium for the propagation of Past.
septica that might yield a luxuriant growth and at the same time be economical.
Several media such as nutrient agar with 0.2 per cent buffer salts (KH 2 PO 4
and Na 2 HPO 4 ), nutrient agar containing 0.1 per cent creatinine, nutrient agar con-
taining 0.5 per cent yeast extract, nutrient agar containing 0.5 per cent yeast extract
and 04 per cent creatinine, nutrient agar containing 0.5 per cent yeast extract,
and 0.1 per cent creatinine and ½ to 1 per cent glucose were tried, and the growth on
each medium was compared. The following medium gave the best growth at our
hands and was, therefore, adopted for the manufacture of the vaccine:
Lab. Lamco.
Peptone Difco
Yeast Extract
Creatinine
Sodium chloride
Distilled water
Agar
.3 gm.
.3 „
1.5 „
0.3 „
1.5 „
300 ml.
9 gm.
Autoclave at 15 lb. for 30 minutes. Final pH 7.2.
(e) Procedure for the manufacture of the vaccine
Stock strain 52 of Past. septica was passaged through a bovine each time a brew
of the vaccine was to be prepared. The organism was recovered from the heart blood
on a blood agar plate and a single colony showing the characteristic appearance was
278
STUDIES
ON
PASTEURELLA SEPTICA
[Vol. XXVI, Part IV,
TABLE II
The results of potency test
Brew No.
1
2
3
No. of
animals
used
in the
experi-
ment
5
31
32
Details of
inoculation
1 ml. subcut
2 ml. subcut
1 ml. S/C. followed
with another dose
of 1 ml. S/C after
12 days
2 ml. subcutaneously
2 ml. intramuscularly
Interval
between
vaccina-
tion and
challenge
21 days
21 days
1 month
2 months
4 months
6 months
1 month
2 months
4 months
6 months
15 days
4 months
5 months
6 months
8 months
Challenge dose
5 million mouse
M. I. D.
do.
do.
do.
do.
do.
do.
do.
do.
5 million mouse
M. I. D.
50 million mouse
M. I. D.
do.
Result
0/2
2/2
2/3
3/3
3/3
2/3
3/3
3/3
3/3
2/3
3/3
5/5
3/5
4/5
3/4
Percen-
tage
survival
(vaccinated
animals)
0
100
66.0
100
100
66.6
100
100
100
66.6
100
100
60
80
75
Reac-
tion
in
controls
0/1
*1/1
0/2
*1/2
0/2
*1/1
0/2
*l/2
0/2
0/2
*l/2
0/2
0/2
0/2
Percen-
tage
survivals
(Controls)
0
100
0
50
0
100
0
50
0
0
50
0
0
0
Result of bacterio-
logical examina-
tion in animals
dying after chal-
lenge
Vaccina-
ted
animals
++
+
+
+
++
+
+
Controls
+
++
+
++
++
+
++
++
+
++
++
++
Remarks
*Recovered
after
showing
severe
illness.
*do.
*do.
*do.
*do.
December, 1956]
M.
R.
DHANDA
ET.
AL.
279
TABLE II—(contd.)
The results of potency test
Brew No.
Brew
No. 4 and
5 (Mixed)
6
7
No. of
animals
used
in the
experi-
ment
35
*2
10
Details of
inoculation
(a) 2
ml.
intra-
muscularly
(b) 4 ml.
intra-
muscularly
5 ml. intramuscu-
larly
3 ml. intramuscularly
Interval
between
vaccina-
tion and
challenge
21 days
2½ months
5 months
6 months
4 months
5 months
21 days
2 months
Challenge dose
50 million mouse
M. I. D.
do.
1,000
million
mouse M.I.D.
50 million mouse
M. I. D.
1,000
million
mouse M. I. D.
50 million mouse
M. I. D.
50 million mouse
M. I. D.
Result
3/5
3/4
5/5
3/4
3/4
4/4
2/2
10/10
Percen-
tage
survival
(vaccinated
animals)
60
75
100
75
75
100
100
100
Reac-
tion
in
controls
0/2
0/2
0/3
0/2
0/3
0/2
0/3*
Percen-
tage
survivals
(Controls)
0
0
0
0
0
0
0
Result of bacterio-
logical examina-
tion in animals
dying after chal-
lenge
Vaccina-
ted
animals
++
+
+
+
Controls
++
++
+++
++
+++
++
+++
Remarks
*Buffalo
calves.
M. I. D. of the
strain was deter-
mined in mice.
No buffalo
calf
was used to serve
as control.
*10 cattle vacci-
nated with broth
vaccine
challen-
ged simultaneously
all died.
NOTE. 1. Adult hill cattle have been used throughout the trials excepting in brew No. 6 where the animals used were buffalo calves about 1½ years in age.
2. Numerators denote survivals. Denominators denote total number inoculated.
3. + = Positive for P. septica on bacteriological examination.
4. A 24-hour old broth culture of virulent P. septica was used for challenge purpose. Of this 1 ml. was determined to contain 100 to 1,000 million mouse M. I. D.
280
STUDIES ON PASTEURELLA SEPTICA
[Vol. XXVI, Part IV,
It will be seen from these results that under laboratory conditions the adjuvant
vaccine in a dose of 2 ml. to 4 ml. intra-muscularly confers adequate immunity in
cattle weighing about 250 lb. for as long as 8 months, the maximum period tested
so far. It may be stated, however, that for the test carried out after 8 months after
vaccination only four animals were available, three of which survived and one died
thereby indicating a survival rate of 75 per cent. Further tests will be carried out in
due course of time to confirm this and to find out the longest period for which animals
vaccinated with this new vaccine would remain immune. The animals were
subjected to a very severe challenge, the dose of the virulent culture used
for the test varying from 5 million mouse M. I. D. to as high a dose as 1,000 million
mouse M. I. D. The virulence of the culture was determined in white mice each
time a challenge dose was to be given.
Trial on the comparative immunogenic value of the H. S. adjuvant vaccine with the
routine commercial broth vaccine
With a view to compare the immunogenic properties of the adjuvant vaccine
with the routine commercial broth vaccine, the following immunity trials were carried
out and the results are presented in Table III.
TABLE III
Comparative immunogenic value of the adjuvant vaccine and the commercial broth
vaccine
Trial
No.
I
II
No. of
animals
used in
the ex-
periment
*23
23
Details of
inoculation
Adjuvant vaccine 2
ml. intramuscularly
Commercial
broth
vaccine 5 ml. S/C
Unvaccinated
con-
trols
Adjuvant vaccine 2
ml. intramuscularly
Commercial
broth
vaccine 5 ml. S/C
Unvaccinated
con-
trols
Adjuvant vaccine 2
ml. intramuscularly
Commercial
broth
vaccine 5 ml. S/C
Unvaccinated
con-
trols
Interval
between
vaccina-
tion and
challenge
2½ months
21 days
-
5 months
3 months and 6
days
—
2 months
Do.
-
Challenge dose
50 million mouse
M. I. D.
Do.
Do.
50 million mouse
M. I. D.
50 million mouse
M. I. D.
Do.
50 million mouse
M. I D.
Do.
Do.
Result
in vac-
cinated
animals
¾
¼
0/2
5/5
0/4
0/3
10/10
0/10
0/3
Percen-
tage
survivals
75
25
0
100
0
0
100
0
0
Results of
bacteriologi-
cal examina-
tion in the
animals
dying
after
challenge
+
+++
++
++++
+++
+++++
+++++
+++
NOTE. Numerators denote survivals. Denominators denote total number inoculated.
+ =Positive for Past. septica on bacteriological examination.
A 24-hour old broth culture of virulent P. seplica was used for challenge purpose. Of this 1 ml.
was determined to contain 1,000 million mouse M. I. D.
*Nine animals were vaccinated with adjuvant vaccine and another group of 9 animals with commer-
cial broth vaccine. Five animals were used as control, One animal of the broth vaccine group was
not available for challenge.
December, 1956]
M. R.
DHANDA
ET.
AL..
281
These results clearly show that the adjuvant vaccine is far superior to the routine
commercial broth vaccine. It will be seen that in trial No. 1 in the case of adjuvant
vaccine, the interval between vaccination and challenge was 2½ months and 5 months
as compared to 21 days, and 3 months and 6 days respectively in the case of broth
vaccine. Thus, the broth vaccine was at an advantage at the time of challenge.
The experiment was planned in this manner, as it was considered that the broth
vaccine was not likely to confer a lasting immunity, and the data obtained would be
more conclusive if the adjuvant vaccine was found superior even when kept at a
disadvantage.
Trial on the comparative immunogenic value of the adjuvant vaccine and Bain's
vaccine with the commerical broth vaccine
During May, 1955, a consignment of Bain's adjuvant vaccines was received at
this Institute for carrying out trials in regard to its safety and potency, for Indian
cattle and buffaloes and in order to determine its efficacy as a prophylactic against
haemorrhagic septicaemia in this country. This vaccine was found to be quite safe
for cattle and buffaloes as well as for young calves and to possess quite good immuni-
zing properties. Opportunity was availed to carry out tests with the Bain's vaccine
and the adjuvant vaccine under study at this Institute to find out the comparative
efficacy of the two products in relation to the old broth vaccine.
The results are presented in Tables IV and V on the basis of trials in white mice
and cattle respectively.
Trial on mice
Three groups each of 10 white mice were vaccinated with the three vaccines and
five mice were left unvaccinated to serve as controls. All the surviving mice were
challenged with a virulent culture of Past. septica after 21 days of vaccination. The
results are set out in Table IV.
TABLE IV
Comparative immunogenic value of the Bain's vaccine and the adjuvant vaccine
with the commercial broth vaccine
No.
of
mice
35
Details of inoculation
0.25 ml. intramuscularly
(Bain's
vaccine)
0.25 ml. adjuvant vaccine
Commercial broth vaccine 0.5 ml. S/C
Unvaccinated controls
Interval
between
vaccina-
tion and
challenge
21 days
do.
do.
-
Challenge
dose
100 M.I.D.
do.
do.
do.
Result
4/5
6/8
2/8
0/5
Percen-
tage
sur-
vival
80
75
25
0
Result of bacteriolo-
gical examination
in the animals dying
after challenge
+
++
++++++
+++++
NOTE. Numerators denote survivals. Denominators denote total number inoculated.
+ = Positive for P. septica on bacteriological examination.
A 24-hour old broth culture of virulent P. septica was used for challenge purpose. Of this 1 ml.
was determined to contain 1,000 million mouse M. I. D.
Ten mice were inoculated with each vaccine but a few died of miscellaneous causes before challenge.
31 AR/56
22
282
STUDIES ON PASTEURELLA SEPTICA
[Vol. XXVI, Part IV,
Trial on cattle
On the basis of age, weight, etc. 30 hill cattle were divided into three equal
groups of 10 animals and each group was vaccinated with one of the three vaccines.
Besides, a fourth group of three animals was left unvaccinated to serve as controls.
These controls were so selected as to represent all the three different age and weight
groups of the vaccinated lots. After two months of vaccination, all the animals
were challenged with ½ ml. of 10-1 of 24-hours broth culture of virulent Past. septica
equivalent to 50 million mouse M. I. D. The results are presented in Table V.
TABLE V
Comparative immunogenic value of the Bain's vaccine and the adjuvant vaccine with
the commercial broth vaccine
No.
of
animals
0
10
10
3
Details of
inoculation
3 ml. intra-
muscularly
(Bain's)
3 ml. intra-
muscularly.
adjuvant
vaccine
5 m. subcu-
taneously
Commercial
broth vaccine
Unvaccinated
Interval
between
vaccination
and challenge
2 months
2 months
do.
do.
Challenge dose
50 million mouse
M. I. D.
50 million mouse
M. I. D.
do.
do.
Result
10/10
10/10
0/10
0/3
Percentage
survival
100
100
0
0
Result of bacteriolo-
gical examination
in animals dying
after challenge
+++++
+++++
+++
NOTE. Numerators denote survivals. Denominators denote total number inoculated,
+ = Positive for P. septica on bacteriological examination.
A 24-hour old broth culture of virulent P. septica was used for challenge purpose. Of this 1 ml.
was determined to contain 1,000 million mouse M. I. D.
The results in mice as well as in cattle clearly indicate that the adjuvant vaccine
under study is as good as the Bain's vaccine in its immunizing properties, and that
both the vaccines are far superior to the routine commercial broth vaccine.
Field trials
Three dairy farms having the history of annual recurrence of haemorrhagic
septicaemia were selected for this study. In two farms, 699 animals
were vaccinated with the adjuvant vaccine and the remaining stock numbering 463
with the commercial broth vaccine. At the third farm, 985 animals were vaccinated
with the adjuvant vaccine, 891 with the Bain's vaccine and the remaining stock
numbering over 15,000 with the commercial broth vaccine.
December, 1956]
M.
R.
DHANDA
ET.
AL.
283
At one of the farms, the records of milk yield in respect of 166 buffaloes and 17
cows vaccinated with the adjuvant vaccine were compared with those of the milk
yield of 93 buffaloes and 16 cows vaccinated with the broth vaccine. It was found
that in both the groups there was an average fall in milk yield of about 0.8 to
1 lb. per animal for three days following vaccination and thereafter the milk yield
returned to normal. This slight decrease in the milk yield is attributed to the inter-
ference with the daily routine of the animals including handling and confinement
to the stalls during this period.
In the field trials, it could neither be possible to leave a few animals unvaccinated
to serve as controls nor to subject a few representative animals of each of the
vaccinated groups to an experimental challenge with a virulent culture, as the farm
authorities were not prepared to take the risk involved. However, it is hoped that
by comparing the figures of mortality due to haemorrhagic septicaemia in the different
groups and also the death rate in the group of the animals vaccinated with the adju-
vant vaccine with the average mortality figures of the past few years, some useful
data regarding the efficacy of the vaccine under study will become available in due
course of time.
SUMMARY
1. The evolution of an adjuvant vaccine against haemorrhagic septicaemia is
reported.
2. Large scale field and laboratory trials have shown that the vaccine is quite
safe for cattle and buffaloes of all ages.
3. Tests have indicated that the immunity conferred by the administration of
the vaccine is lasting for at least eight months against a heavy challenge with virulent
P. septica. The results in respect of 8-month period, however, require to be confirmed
on a larger number of animals as only four animals were used for challenge of which
3 (75 per cent) survived.
4. On comparative trials, this vaccine has been found to be definitely superior
in its immunising properties to the commercial broth vaccine at present issued from
the Institute.
ACKNOWLEDGMENT
The authors are thankful to M/s. T. P. Banerjee, P. Chandra Sakariah and M. S.
Sethi for technical assistance and to Shri S. Sen for compiling the figures given in
Table I. They also thank Brigadier H. L. Bhandari, Col. Nayyar and Capt.
W. H. Dean of the Remount, Veterinary and Farms Corps, Mr. D. N. Khurody,
Milk Commissioner, Bombay and M/s. M. R. Marathe and N. R. Adker of the Aarey
Milk Colony for their co-operation in field trials.
284 STUDIES ON PASTEURELLA SEPTICA [Vol. XXVI, Part IV,
REFERENCES
Annual Report of the Pathology and Bacteriology Section (1942-43), Indian Veterinary Research
Institute
Bain, R. V. S. (1953). Personal communication
Cornelius, J. T. (1929). An investigation of the serological relationships of twenty-six strains of
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Fitch, C. P. and Nelson, E. N. (1923). Prelim nary report on the differentiation of various
organisms belonging to the haemorrhagic septicaemia group. J. Amer. Vet. Med. Ass. 63, 147
Jones, F. S. (1921-22). The types of Past. bubaliseptica in a dairy herd. J. Amer. Med. Ass.
63, 271
Khalifa, A. B. (1936). Pasteurella in animals and their inter-classification. Ministry of Agricul-
ture, Egypt. Technical and Scientific Service, Veterinary Section, Bulletin No. 147. Quoted by
Rao, U. R. K. (1953). Studies on Pasteurella septica. Thesis: University of Madras
Lal, R. B. (1927). A study of certain organisms of the Pasteurella group with respect to specific
complement fixation by the ice box method. Amer. J. Hyg. 7, 561
Rajagopalan, V. R. (1944). Haemorrhagic septicaemia in buffaloes and cattle. Indian Fmg. 5,
No. 4, 176
Rao, U. R. K. (1953). Studies on Pasteurella septica. Thesis: University of Madras
Roberts, R. S. (1947). An immunological study of Past. septica. J. Comp. Path., 57, 261
Roderick, L. M. (1922).
Some antigenic relationships
of Bipolaries septicus group of
bacteria. J. Inf., Dis. 31, 313
Scientific report of the Division of Pathology and Bacteriology of the Indian Veterinary Research
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Yusef, H. S. (1935). A contribution to the serological classification of Pasteurella strains.
J. Path. and Bact. 41
ABSTRACTS
A comparison of the intradermal and subcutaneous routes in producing immunity
to brucellosis in cattle. McDIARMID A. (1954). J. Comp. path. 64, 384-91
T HE author has compared the results of immunity produced in groups of ten
heifers vaccinated with 'Cotton strain 19' at the dose-rate of 0.2 ml. intradermal-
ly, 0.2 ml. subcutaneously and 5.0 ml. subcutaneously. A batch of ten unvaccinated
heifers was kept as controls. All the heifers were obtained from a non-vaccinated
brucellosis-free herd. At the time of vaccination they were in calf for two months
and negative to agglutination test for brucellosis. The reaction after vaccination by
intradermal route was the production of a small local necrotic lesion which rapidly
healed. Whereas slight oedematous area was observed in 0.2 ml. subcutaneous dose,
5.0 ml. dose exhibited usual systemic and local reactions. Serum samples from
vaccinated heifers were tested for agglutinins from time to time. After three months
of vaccination, all the animals were challenged by instilling into conjunctival sac
1 ml. of virulent Brucella abortus strain 544 containing 134 million live organisms.
At parturition, serum titre was recorded and cotyledon foetal stomach-contents,
colostrum, milk samples were examined culturally and biologically. The number
of calves that were born full-term and survived after challenge in 0.2 ml. intradermal
group, 0.2 ml. subcutaneous group and 5.0 ml. subcutaneous group was 6, 7 and 7
respectively, while in the unvaccinated controls all the calves were born dead. The
immunity produced by 0.2 ml. intradermally was found to be equal to that obtained
by the same quantity of vaccine given subcutaneously. However, in both of the
0.2 ml. groups and in the group vaccinated with 5.0 ml., the level of immunity was
poor since 5 out of 10 animals were found infected in each group after challenge.
These results are at variance with the previous findings of the author. The factor
or factors responsible for such discrepancies are not known, but in the opinion of the
author the possibilities are (a) that a pregnant heifer does not respond so well to the
Brucella abortus antigen as does the non-pregnant heifer and (b) the short interval
between immunization and challenge. The mode of administration of a correct
challenge dose requires careful consideration whenever such immunity experiment
are performed. The author affirms that vaccination of pregnant cattle with strain
19 vaccine can be carried out without any risk of abortion. The finding is of great
importance when dealing with an abortion storm in a non-vaccinated herd (P.C.).
Antibiotics and nitrogen utilization in growing cockerels. R. H. THAYER AND
V. G. HELLER (1955). Poult. Sci., Vol. 34, No. 1, pp. 97-102.
E XPERIMENTS were carried out to determine the relative efficiency of protein
utilization between antibiotic and non-antibiotic for chicks using both growth
response and nitrogen balance techniques. The study consisted of four feeding
285
31 AR/56
23
286
ABSTRACTS
[Vol. XXVI, Part IV,
trials conducted in battery brooders with the basal diet supplemented with 20 mg.
per lb. of either procaine penicillin or aureomycin hydrochloride. The length of the
feeding period varied from 27 to 33 days in the different trials. The nitrogen balance
study was carried out with four-week old cockerels to determine the effect of penicillin
and aureomycin upon dietary nitrogen utilization.
It was observed that the net result of antibiotic feeding was to produce greater
growth and more efficient feed conversion. The results indicated that penicillin and
aureomycin brought about an increase in the amount of dietary nitrogen which was
digested and absorbed from the digestive tract of the four-week old cockerels. A
possible explanation is that the rate of digestion may have in some way been increased.
The antibiotics may have reduced the number or type of competitive intestinal
bacteria which used amino-acids as they were released in the digestion process.
The data indicate that the increased growth response due to the addition of anti-
biotics to rations for chickens may be due to an increased efficiency in the digestion
and absorption of dietary nitrogen. (S. B.).
The effect of sexual stimulation prior to service on the behaviour and conception
rate. B. M. KERRUISH. Brit. J. Anim. Behaviour. Vol. III, No. 4, pages
125-130.
I N this article the author has mentioned the behaviour of an. experienced bull
running with the herd at precoital, coital and postcoital state. He has also
mentioned the behaviour of bulls due to incomplete sexual stimulation under con-
ditions of controlled mating and collection of semen by artificial vagina. Sex drive
and vigour of the ejaculatory reflex are significantly lowered in following the rigid
routine of controlled mating in most artificial insemination centres and farms. The
bulls fail to get sufficient precoital excitement, which probably not only affect
sexual organs but also plays a part in promoting the general metabolism of the
body through the levels of secretion of endocrine glands, thus increasing the excita-
bility and tone of the whole body in giving a vigorous and better co-ordinated ejacula-
tion.
The experiments revealed a significant rise in conception rate with increase in
precoital stimulation. Animals not getting sufficient excitement prior to coitus
appear to be disinterested and unresponsive to service and do not give collection
readily. The fall in conception rate may be due to the poor quality of semen pro-
duced. (D. P. M.).
Familial resistance to Newcastle disease in strain of New Hampshires. FRANCIS,
D. W. and A. F. KISH (1955). Poult. Sci. 34, 2, 331-336.
I N this paper an attempt has been made to determine genetic differences among
six family lines of New Hampshire chickens in respect of resistance to a standard
challenge dose (LD 50) of Newcastle disease virus. Birds used in mating to raise
chicks for challenge were selected from different families having mortality varying
from 3 per cent to 17 per cent. The family differences in mortality rate following
the challenge were used to evaluate the degree of resistance.
Vol. XXVII, Part I
March 1957
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