ICMR BULLETIN VOL.8-No.-8-AUGUST-1978
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- ICMR BULLETIN VOL.8-No.-8-AUGUST-1978
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Vol8 No.8
COMMUNIT VRLrHleEe
326, V Main, I Block
VIRUS
RESEARCH
(1977-78)
Koramongala
/
Bangalore- 560034 *
Virus
diseases like influenza, polio,
hepatitis,
encephalitis and dengue occur in
major epidemics in our country. Many of these
diseases
exclusively
affect
agricultural
labourers working in the forests,
as in the
case of Kyasanur Forest disease (KFD)
in
Karnataka.
In other instances like Japanese
encephalitis (JE),
the disease is known to
effect people living in villages in close
proximity with pigs and fowl.
There are no
drugs effective against these virus diseases.
Prophylactic control measures alone, based on
a knowledge of the virus and virus vaccines
can help in protecting human beings against
these diseases.
The Virus Research Centre (VRC), Pune,
which has been recognised as a premier centre
in our country for research on arboviruses has
now been renamed as National Institute of
Virology (NIV), in view of its expanding acti
vities in the field of virology.
Since its
inception the NIV has been carrying out major
surveys in an effort to look for evidence of
infection by viruses.
These surveys have
taken the NIV field teams to far flung areas
of the country from Kashmir to Kerala and from
Arunachal to Kutch. A large part of India has
thus been mapped with regard to the existence
of viral and rickettsial activity.
JAPANESE ENCEPHALITIS
Since the appearance of Japanese ence
phalitis (JE) in West Bengal in 1973, NIV has
been continuously engaged in extensive studies
in this area.
During the year, a JE-like out
break was reported from Chintamani taluk near
Bangalore.
The field station of the NIV at
Bangalore is currently investigating this out
break.
Antibodies to JE virus are found exten
sively distributed in all parts of the coun
try.
While the role of pigs in the mainte
nance of the disease has been established, the
probable role of birds and several species of
mosquitoes in the natural cycle of JE virus
has been receiving attention. Several strains
of JB virus have been isolated from mosquitoes
at Bankura.
Studies have been carried out .on
the vectorial capacity of C. bitaeniorhynchus
and An. hyrcanus mosquitoes.
It was found
that C. bitaenioi4iynchus could transmit JE
virus effectively.
Though no transmission
could be demonstrated by An. hyrcanus^
the
virus was found to persist in the mosquitoes
upto 11 days after the infective feed.
Studies on birds have indicated that
ardeid birds develop viraemia and a bird-mosquito-bird cycle has been demonstrated under
experimental
conditions,
suggesting
that
ardeid birds may play a role in the natural
cycle of JE virus.
During the year 55 per
cent of 196 wild ardeid birds caught in Andhra
Pradesh showed neutralizing antibodies to JE
virus. Experimental infection in ardeid birds
produced specific neutralizing antibodies to
JE virus.
Experimental studies on ducks are
in progress.
Pigeons were found to circulate
high, levels of JE virus suggesting that a
large proportion of vector mosquitoes fed on
them could be infected.
The possibility of parasite-virus inter
action leading to the occurrence of a disease
NIV for the past few years.
During the year,
experimental work was carried out in pigs by
infecting them by the oral as well as the sub
cutaneous routes.
It was found that JE virus
could infect the pig through the oral route,
particularly when the pigs were also infected
with parasites. Unlike in mice,
there was no
significant sickness or death in pigs either
with the virus alone or in combination with
helminths.
The ICMR Virus Research Unit at Calcutta,
investigated an outbreak of JE which occurred
in 1976, in collaboration with the NIV, Pune,
and the Vector Control Research Centre, Pondi
cherry.
In this outbreak a total of 277
clinical cases were reported.
Specimens of
acute blood (215),
convalescent blood (76),
CSF (90), postmortem brain (81),
cattle sera
(7), pig sera (47),
goat sera (4) and 1475
mosquitoes from the affected area were re
ceived for investigation.
JE virus could be
isolated from a mosquito pool (C. vishnui) and
a postmortem brain sample.
Of the 76 paired
sera tested by HI and CF tests,
33 showed a
rise in titre against JE virus and 32 against
Group B viruses while 11 did not show any
definite rise in the titre.
Of the 47 pig
sera,
22 showed a high titre of HI antibody
against JE virus.
The cattle and goat sera
were negative against JE antigen.
The Unit also continued studies on the
production of a vaccine against JE using a
local strain of virus in tissue culture.
KYASANUR FOREST DISEASE
Since the discovery of Kyasanur Forest
disease (KFD) in Karnataka State by the Insti
tute in 1957, the NIV has been responsible for
monitoring the activity of the virus in the
KFD area. Since 1975,
although the Karnataka
State Government has taken over the responsi
bility of surveillance of the activity of the
KFD virus,
the NIV continues to help the
State’s diagnostic laboratory in identifying
the virus strains.
Improvement in the existing formalized
KFD vaccine was attempted by using Roux bottle
cultures of chick
embryo fibroblasts and
concentrating the virus by precipitating it by
polyethylene glycol and by using aluminium
hydroxide gel as an adjuvant.
Concentration
of the virus from pools of mouse brains could
also be obtained by treatment with poly
ethylene glycol.
Studies have been initiated on the pre
paration of a sub-unit KFD vaccine.
Brains
2
from infected mice are being utilised for this
purpose, employing SDS, -2-mercapto ethanol,
tween 80 and ether and triton X-100 for break
ing up the virus.
Studies on mice with sub
unit vaccine have indicated the possibility of
developing such a vaccine for use in human
beings.
Efforts to develop an attenuated KFD
vaccine were continued. A strain of KFD virus
was passed in primary chick cell cultures.
Comparison between titres of the virus in the
mouse brain, for the candidate strain for KFD
vaccine and the wild strain indicated that
while the candidate strain multiplied,
it did
not develop any CF antigen.
Further studies
are in progress.
Studies on the role of birds in the
maintenance and spread of KFD virus have
yielded interesting results.
A total of 127
serum samples from 10 migrating and 7 resi
dent species of birds from the Bharatpur bird
sanctuary collected in 1973 were tested for
antibodies to several Group A and Group B
viruses as well as Congo and Batai viruses.
Fifteen sera from four species of migratory
birds were positive for HI antibodies to one
of the Group A or B arboviruses.
Only one
showed HI antibodies to KFD while six had ’N*
antibodies to KFD virus.
Studies on tick vectors of KFD were
continued.
At least 15 species of ticks are
known to transmit KFD virus in nature. One of
them is Haemaphy salts papuana ktnneart.
Life
history of this tick in the laboratory was
studied during the year.
Studies were conti
nued on the tick fauna,
as related to their
preference to vegetation,
vertical distri
bution in the questing stages and the differ
ent hosts on which various stages of the ticks
feed, and the size of the preferred host.
Colonization of C.
fuscocephalus
and
C. whitmorei has been attempted in order to
study their vectorial
capacities for KFD
virus.
Studies on tick repellence using 6 chemi
cals were carried out with 6 species of Ixodid
ticks. A new method for the evaluation of re
pelling property of such chemicals has been
worked out.
DENGUE
Dengue is known to occur throughout the
country and all the four types of dengue virus
I
have been isolated.
During the year,
several
cases of pyrexia of unknown origin in Pune
were investigated by the NIV and 20 per cent
of them turned out to be cases of dengue. NIV
is also carrying out studies on the trans
mission of dengue by mosquitoes maintained on
chicken and human blood.
Studies carried out
by the NIV on cases of acute paraplegia in
collaboration with Ruby Hall Nursing Home,
Pune, have indicated that dengue virus infec
tion could be responsible for some of the
cases of Guillain-Barre syndrome and acute
transverse myelitis.
control of mosquitoes. The NIV is now testing,
in collaboration with the National Chemical
Laboratory,
Pune,
a series of compounds
against
the
fourth
instar
larvae
of
Ae. aegypti.
The pathogenesis of dengue haemorrhagic
fever (DHF) and dengue shock syndrome (DSS),
which are rare in India,
has not been under
stood so far, in spite of large amount of work
carried out in many parts of the world.
It
has been suggested that IgE-mediated release
of histamine followed by increased vascular
permeability
may be responsible
for the
occurrence of DHF and DSS.
IgE level was
therefore determined in a few sera from the
1958 outbreak of DHF in Thailand, and IgE was
found to be significantly higher in them com
pared to the controls.
During the year,experimental studies were
carried out on Wanowrie virus infection in
adult and infant mice.
Virus titrations were
carried out on various tissues and organs of
experimentally infected mice during the first
few days post-inoculation (PI).
Virus titres
varied in different tissues. Lesions in adult
mice were confined to the liver showing marked
vacuolation of the liver cells with small
patchy areas of necrosis. The vacuolation was
due to fatty change.
There was proliferation
of the endothelial cells lining the hepatic
veins.
Lungs also showed interstitial pneu
monitis and haemorrhages. The small intestine
of infant mice showed congestion and necrosis
of the mucosa.
No pathological lesions could
be detected in the brain.
Hie ICMR Virus Research Unit at Calcutta,
has been actively engaged in surveillance of
dengue, particularly DHF, in West Bengal.
No
frank case of DHF has so far been encountered.
Of the 143 cases of pyrexia of unknown origin
studied, in four dengue virus could be iso
lated. Serological survey for arbovirus anti
bodies in different parts of West Bengal were
continued.
Preliminary experiments undertaken by the
Virus Research Unit,
Calcutta,
to produce
features similar to human DHF in mice, using
dengue virus type 1,
as the initial infecting
virus,
and adult adapted dengue virus type
2(AD-2) for secondary infection have yielded
promising results.
Further studies are in
progress to investigate if passive transfer of
pre-formed antibodies produces the same pic
ture as previous infection.
Studies are also being carried out by the
NIV, Pune, on the mosquito vector of dengue
i.e. Aedes aegypti, in Maharashtra State.
As
a first step in the study of strain variation
in Ae. aegypti* a strain maintained in the NIV
colony has been screened by MacClelland’s
graded series of abdominal variants.
Further
screening is in progress.
The use of insect growth regulators has
been suggested as one of the many methods for
WANCWRIE VIRUS
Wanowrie virus is one of the new viruses
isolated by the NIV from HyaLamma marginatum
isaaci collected from sheep near Pune.
There
is report of only one fatal case caused by
this virus.
CRIMEAN HAEMORRHAGIC FEVER (CHF) - CONGO VIRUS
The NIV has been engaged during the last
two years in extensive studies to determine
the prevalence of the activity of CHF-Congo
virus in the country.
Intensive serological
survey of human beings,
cattle, sheep, goats,
mules, rodents and birds is being conducted
specially in the northern parts of India.
A
large number of ticks are also being processed
for virus isolation.
GANJAM VIRUS
Studies on Ganjam virus were continued by
the NIV.
Pathological changes studied in in
oculated mice indicated that the virus causes
lesions in the liver, brain,
lung and kidney
of the inoculated mice.
The NIV investigated an "ill-defined”
disease in local,
exotic and cross-bred sheep
in Palamer
of Chittoor district,
Andhra
Pradesh.
Acute phase serum samples from
sheep and tick pools of Haemaphysatis inter
media yielded
Ganjam virus.
Sera
were
collected from local as well as exotic and
cross-bred sheep.
This is the first report
3
from India of a disease in sheep caused by
Ganjam virus, which resembles the Nairobi
sheep disease virus.
botomus fever viruses were used in chicken and
some inhibitory effect was noticed.
Further
studies are in progress.
SEQUENTIAL IMMUNIZATION
The role of extra-human vertebrates in
the natural cycle of phlebotomus fever virus
is not fully understood. More than 20 species
of laboratory-bred and wild-caught specimens
have been tested and virus circulation could
be detected in squirrels only.
Hamsters bred
in the laboratory were also found to yield
peak titres.
Group B arboviruses viz. JE, KFD, dengue,
West Nile, etc.,
are widely prevalent in our
country. Repeated infections are not unknown.
Sequential infection by dengue viruses is
alleged to be responsible for the occurrence
of DHF/DSS.
Individuals with antibodies to
these viruses prior to infection may not some
times show any antibody rise after vaccina
tion.
Further studies are required on the
mechanism of cross-protection and cross-reac
tion and pathogenesis of Group B arbovirus in
fection.
The NIV has been attempting to find
out if sequential immunization with one or
more viruses of this Group can offer protec
tion against infection by a large number of
Group B arboviruses.
Whilst monkeys had been
used in the earlier studies at the Institute,
currently rabbits are being used.
Four doses
of the viruses in various permutations and
combinations of JE, WN, dengue-2,
and KFD
(attenuated, killed and virulent) viruses have
been employed.
The results of this study are
now being analysed.
Studies are also being carried out on IgG
and IgM antibodies
and their
sequential
development in infected rabbits and cross-reaction between these two antibodies.
Studies
are in progress to detect circulating immune
complexes in rabbits sequentially immunized
with different flaviviruses.
Experimental studies in the laboratory on
the role of P. papatasi have been undertaken
to understand the rate of engorgement,
infec
tion, transmission, persistence and multipli
cation of the Naples and Sicilian strains of
phlebotomus fever virus.
CHICKENPOX
Attempts are being made to develop a
chickenpox vaccine. As the virus could not be
grown satisfactorily in Vero culture, addition
of a steroid/DEAE dextran to the tissue cul
ture is being tried.
HEPATITIS
The NIV investigated an outbreak of hepa
titis in Nellore district of Andhra Pradesh in
February, 1977. An interesting aspect of this
outbreak was its simultaneous occurrence in
25 villages.
The NIV studied 230 samples of
sera for the presence of HBsAg and for the
distribution of antibodies. HBsAg was detected
in the urban as well as the rural areas.
SANDFLY FEVER
Sandfly fever is known to have reappeared
in India. Both the Naples and Sicilian strains
of the virus were identified by the NIV in an
outbreak near
Aurangabad some years ago.
Attempts have been made to plaque this virus
in LLC MK-2, Vero and PS cell lines using
Linbro plates.
It was demonstrated that
phlebotomus fever virus could grow in monkey
kidney cultures with high titres but without
producing any CPE while in PS cell line,
the
CPE was noted from the 4th day onwards.
Attempts were made to produce haemagglutinin
of phlebotomus fever virus.
No haemaggluti
nation was demonstrated by using tissue cul
tures procedures but infected mouse brains
showed its presence on the 6th day after in
oculation.
Many arboviruses are known to produce in
hibition of growth of Rous' sarcoma.
Phle
4
Attempts have been made by the NIV to
standardise IAHA for detection of HBsAg and
also to find out a suitable human *0' group
donor for this purpose. Purification of hepa
titis A virus from faecal material has also
been attempted by initial extraction,
concen
tration with polyethylene glycol,
followed by
passage through Sephadex 2-B column.
At the ICMR Virus Research Unit,Calcutta,
blood samples from 492 healthy individuals and
the same number of voluntary blood donors were
examined for detection of Australia antigen.
It was found that 2.8 per cent of the appa
rently normal healthy population and 4.1 per
cent of the voluntary blood donors in Calcutta
carry this antigen. The incidence appeared to
be higher in male adults than in females.
There was no significant difference in the
percentage of positive carriers in different
age groups.
The Australia antigen incidence
in the normal population of Calcutta (2.8 pei
cent) seemed to be similar to that of Delhi
and Chandigarh (2.7 per
(2.76 per cent),
cent).
POLIOMYELITIS
Comparative studies on the killed paren
teral vaccine and live oral poliovaccine are
being continued at the Department of Microbio
logy, All India Institute of Medical Sciences,
New Delhi.
The object of this study is to
find out the method most suitable for immuniz
ing Indian children against poliomyelitis.
Attempts were made to investigate if the res
ponse to live poliovaccine given orally could
be improved by
(a) increasing the number of
doses,
(b) altering the interval between the
doses, and (c) changing the time of the year
for administration of the vaccine.
Prelimi
nary results suggest that increasing the num
ber of doses might improve the response. Fur
ther studies are in progress.
Another study is in progress at the B.J.
Medical College, Ahmedabad,
to determine the
schedule, number of doses and the quantity of
poliovirus in each dose of oral poliovaccine
in order to achieve an effective sero-conversion and 100 per cent protection
against
poliomyelitis in children.
A study was also carried out on the ex
cretion
of poliovirus in
children after
administration of oral poliovaccine.
In 142
children from whom
faecal specimens were
collected and examined,
83 (58 per cent) ex
creted viruses in the stool after the first
dose of oral poliovaccine,
48 (34 per cent)
after the second dose,
and 35 (25 per cent)
after the third dose.
About 24 per cent
children did not excrete any virus in their
stool after any dose of oral poliovaccine.
Higher rate of excretion of the virus was
found after oral poliovaccine administration
in the age group 0-12 months, while the rate
declined in the age group 13 to 36 months.
Poliomyelitis surveillance was continued
at the Council’s Enterovirus Unit at Bombay.
During January - November 1977, 986 cases of
paralytic poliomyelitis were recorded,
30 per
cent of the total cases were recorded between
May and August.
During
the
year,
paralytic illness
following
history
of
oral
poliovaccine
administration was recorded in 206 cases.
Of
these, 131 cases were subjected to virological
studies and 98 were found to excrete an in
testinal virus.
Polio virus was detected in
90 cases, the predominant strain being type 2.
A 60 day follow-up study was carried out in
539 cases of paralytic poliomyelitis to assess
residual paralysis.
It was found that while
1 1.5 per cent cases had no residual paralysis
and 77 per cent showed partial recovery of
paralysis, in about 12 per cent cases there
was no recovery at all.
During poliomyelitis
surveillance, approximately 10 per cent of the
non-vaccinated group of population was studied
for virus excretion. Of the 67 cases, 59 were
found to excrete intestinal viruses.
In 52
cases, there was poliovirus excretion, type 2
being the predominant strain, in contrast to
the earlier isolations.
The Enterovirus Unit at Bombay, has ini
tiated a programme to test the potency of oral
poliovaccine at the time of its actual use in
the field.
For this purpose, weekly samples
have been collected from a number of immuniza
tion centres from different parts of the city.
About 260 samples have so far been collected
and tested.
About 93 per cent of the field
samples of oral poliovaccine have not shown
any drop in the potency.
The study is still
in progress.
The Unit is also assessing the
long-term stability of the oral poliovaccine
samples from a large number of vaccine lots
stored at -20 °C for a period of 2 years at
the Haffkine Institute, Bombay. Approximately
6 per cent of the vaccine lots have shown
significant drop of potency
on prolonged
storage.
In continuation of studies on inhibitors
of poliovirus in human milk and faeces, the
Unit has now started collecting samples of
saliva from infants. The aim of this study is
to assess if there are any poliovirus inhibi
tors in the saliva which may adversely influ
ence the activity of the virus in oral polio
vaccine. So far, 25 samples have been collec
ted and tested. None of the samples has shown
the presence of inhibitors.
In view of the disturbing seroconversion
rates obtained from trivalent oral polio
vaccine,
the Enterovirus Unit at Bombay has
planned to immunize children with monovalent
oral poliovaccine. The schedule that is being
followed is monovalent type 1 followed by type
3, followed by type 2, followed by trivalent
vaccine. Of the 96 infants who have been sub
jected to this study so far, 24 have completed
the full schedule of the immunization.
Fur
ther studies are in progress.
5
INFLUENZA
The NIV has been engaged in studies to
isolate influenza virus and survey the anti
bodies in human sera. Studies are also under
taken on animal and bird sera collected in
large numbers at the Institute.
A number of throat swabs from suspected
cases of influenza collected from the Pune
Municipal Corporation dispensaries were pro
cessed and 17 haemagglutinating agents were
isolated in 1976 and nine agents in 1977.
Some of these strains were identified as being
related to A/Victoria/3/75.
Most of these
strains were highly sensitive to noh-specific
inhibitors in chicken serum.
Studies on the antibodies in animals to
influenza A virus were continued.
During the
year, more than 300 goat sera were tested and
most of them were found to be negative.
More
than 100 cloacal swabs which were
collected
from pigeons from the Zoological garden in
Pune have been processed for virus isolation.
RABIES
Rabies continues to be a serious problem
in the country.
The number of persons who
undergo antirabies treatment is steadily in
creasing every year.
The Council is support
ing several investigations on rabies and the
important aspects of the disease which are
being studied at different centres in the
country are
epidemiology,
diagnosis
and
improvement of the vaccine.
A study is in progress at the B.J. Medi
cal College, Ahmedabad, to determine the inci
dence of carrier state among stray dogs. There
was no evidence of a carrier state among the
animals examined so far.
The immunoperoxidase test used in the
diagnosis of rabies has the advantage of
detecting the antibody with an ordinary micro
scope in contrast to the fluorescent antibody
test. Studies at the Central Research Insti
tute, Kasauli, have shown that the test is
satisfactory in cases of brains with a high
concentration of rabies virus but is not
dependable when the virus concentration is
low.
Conventional rabies vaccine contains a
large amount of nerve tissue and the encephalitogenic factor present in the
nerve
tissue may lead to the occasional development
of neuroparalytic accident in treated persons.
6
The vaccine could be
rendered relatively
free of encephalitogenic factor if the brains
of very young animals are used for the pre
paration of the vaccine.
But the best method
would be to prepare a vaccine with the virus
grown in tissue culture.
Another method of purification that could
be adopted is treatment with the fluorocarbon,
arcton 113,
which selectively precipitates
the nerve tissue which contains the encephali
togenic factor.
Studies at the Central Re
search Institute, Kasauli, have shown that by
arcton treatment the protein content of the
brain emulsion could be reduced to about a
fourth,
while the
infectivity was
only
slightly reduced.
The vaccine lots prepared
from arcton treated infected brain suspensions
were found to pass the potency test.
Another study employing arcton to reduce
the nerve tissue protein was carried out
at
the National Institute of Communicable- Dis
eases, Delhi. In this investigation the brain
emulsion was sonicated to improve the virus
titre and then treated with arcton.
The
vaccine lots prepared have passed the potency
test.
FESPIPATORY SYNCYTIAL VIRUS
Respiratory syncytial virus (RSV)
is an
important virus causing acute respiratory illness.
In children, RSV is known to cause
acute bronchiolitis, bronchitis and bronchopneumonia whereas in adults it causes colds
and minor respiratory
disorders.
At the
Institute of Microbiology, Madras,
throatswabs from 400 patients in the age group 0-30
years,
suffering from lower and upper respiratory infections, were inoculated into human
amnion tissue cultures for the isolation of
RSV.
A total of 8:2 strains producing CPE
characteristic of RSV were isolated, of which
75 were confirmed as RSV.
Children up to
5 years of age were found to suffer from a
variety of lower respiratory illness due to
RSV, the most serious cases occurring in
children below one year of age. After the Sth
year, the incidence of recovery of RSV from
serious lower respiratory disorders declined,
although the rate of recovery of RSV from
minor lower respiratory and upper respiratory
infections was found to be on the increase.
On screening the sera of 544 apparently normal
individuals in the age group 0-50 years for
CPF antibodies against this virus, 23 per cent
of the population were found to have had past
experience with the virus,
the highest per
centage (27.5 per cent) being seen in the age
group 0-10 years.
HERPESVIRUS
In addition to the screening of the sera
of oropharyngeal cancer patients,
the ICMR
Virus Unit at Ahmedabad carried out studies on
patients of cervical cancer alongwith matched
controls.
After collecting epidemiological
data,
438 blood samples were tested by IHA
test with herpes virus type 2 antigen, while
59 sera were tested by complement fixation
test, and 47 by neutralization.
In nearly 80
per cent of the cases of cervical cancer,
antibodies were found for herpes virus type 2
by IHA as compared to 35 per cent in the con
trol group,
giving a significant difference
in the percentage positivity.
Immunological studies on herpes simplex
virus have been initiated at the All India
Institute of Medical Sciences, New Delhi,
with an aim to compare IHA test and radio
active chromium (51Cr) release test for their
suitability in the study of the seroepidemiology of herpes simplex virus (HSV) infections.
The two tests have now been standardised and
studies are in progress to evaluate them in
cases of cervical cancer,
oral and genital
herpes infections,
and normal healthy con
trols.
CLINICAL VIROLOGY
At the Clinical Virology Unit of the NIV,
Pune,
165 cases of pyrexia of unknown origin,
suspected cases of encephalitis,poliomyelitis,
chickenpox,
viral myocarditis, rabies, etc.
were investigated.
Paired sera from 126
patients and a specimen of brain for autopsy
were obtained.
Nearly 20 per cent of the
cases investigated were due to dengue. Influ
enza A virus infection was diagnosed in 8
cases and Gan jam virus was isolated in 5
patients. Haematological investigations showed
a low platelet count in 4 patients with a pro
bable diagnosis of dengue.
The NIV investigated the aetiology of a
large epidemic of gastroenteritis in some
areas of the Ratnagiri district of Maha
rashtra.
In this outbreak, 4,000 cases of
gastroenteritis/dysentery were reported, 50 of
them being fatal.
Bacteriological studies
carried out elsewhere on 52 stool samples had
revealed the prevalence of Shigella infection
in about one-third of these cases. NIV tested
61 stool samples for rotavirus
by counterimmuno-electrophoresis and electron
micro
scopy.
Seven of the 61 samples showed preci
pitation lines for rotavirus.
Electronmicroscopic study, however, did not reveal any
rotavirus particles, but showed coronaviruslike particles in 10 samples and enterovirus
particles in one.
CHEMOTHERAPY OF VIRUS DISEASES
The NIV has initiated studies on chemo
therapy of virus diseases including studies on
interferon, in collaboration with the National
Chemical Laboratory (NCL), Pune,
and the
Central Drug Research Institute (CDRI),
Luck
now. 6-MFA, an interferon inducer, was iso
lated from Aspergtltns ochraceus by CDRI,Luck
now. 6-MFA was more effective against viruses
like West Nile and KFD and has found to be
different from the well known interferon in
ducer Statalon
(obtained from Penicillium
stotoniferum).
Synthetic
chemotherapeutic
agents supplied by the NCL, Pune, are also
being tested against some viruses in the labo
ratory .
TISSUE CULTURE AND CELL BIOLOGY
Tissue culture and cell biology studies
at the NIV are concerned with two major areas:
firstly, studies are carried out on the effect
of viruses on cell lines, cell organelles and
organ cultures and secondly, efforts are made
to characterise the normal and transformed
cells.
Of the 46 cases of Guillain - Barre syn
drome and 35 cases
of transverse myelitis
studied, 40 and 31 cases respectively showed
antibodies to one or more viruses.
Studies on the effect of viruses on
chromosomes were continued with measles virus
in human blood leucocyte cultures and hamster
cell cultures. Chromosomal aberrations obser
ved included chromatic breaks, gaps, frag
mentation and pulverisation.
High titres of antibodies were found
against six Group B arboviruses,
one against
measles virus,
and four against herpes virus.
Of the 11 cases of Guillain - Barre syndrome,
9 were due to Group B viruses, one each due to
measles and herpes virus.
The possibility of
the involvement of WN and dengue-2 has also
been considered in some of these cases.
Tracheal organ cultures were prepared
from reptiles, birds and mammals and several
viruses, particularly Sendai, measles, chikungunya, picorna, myxo and arboviruses were
tested.
DNA viruses included vaccinia and
herpes viruses.
The results indicated that
mammalian tracheal explants supported multi
plication of vaccinia, herpes simplex type 1
7
and Sendai viruses by exhibiting ciliostasis
and of Chandipura, West Nile, herpes simplex
type 2, polio and sandfly viruses without any
noticeable effect on the ciliary activity. JE
and chikungunya viruses did not grow in these
cultures.
On the other hand,
chick embryo
trachael culture supported multiplication of
JE and chikungunya viruses.
Interference
phenomenon could be demonstrated in tracheal
cultures by observing ciliostasis.
Attempts
were also made to locate the site of virus
multiplication in tracheal
organ cultures
using autoradiographic techniques.
Studies were continued on organ cultures
of liver and kidney.
It was shown that both
could support the growth of a number of
viruses. Studies on characterisation of cells
involving the behaviour of normal virus infec
ted and transformed cells in the presence of
Concanavallin A (Con A) were also carried out.
The studies on the agglutinability of differ
ent experimental cell cultures with Con A were
continued.
Normal diploid cells did not
agglutinate with Con A.
But cells infected
with viruses agglutinated before visible CPE
was observed.
Chromosomes in insect cells were also
studied in vitro.
Studies have been carried
out by autoradiographic technique on cell
lines obtained
from
Ae. albopictus,
Ae.
w-albus and Ae. novo albopictus.
Attempts
have been made to understand banding patterns
in chromosome formation.
These chromosomes
were also studied after infecting the cells
with JE, WN and herpes simplex and measles
viruses. Chromosome studies of the virus in
fected mosquito cell lines
revealed that
despiralization of the metaphase chromosome
was a major effect besides chromatid and
chromosome breaks with JE and WN viruses.
Other changes in chromosomes are under study.
Characterization of insect cell in vitro
was attempted on a number of cell lines,
specially- Ae. novo albopictus and Ae. vittatus
and Ae. y-albus.
Studies have been carried
out on the morphology and growth rate includ
ing estimation of cellular proteins
Histo
chemical studies specially with regard to
esterases and lipids,
microspectrophotometric
studies on cellular DNA and enzyme patterns of
these cell lines are also being carried out.
It was possible to differentiate various mos
quito cell lines by their characteristic LDH
and MDH electrophoretic patterns.
The pioneering work of the NIV on Singh’s
Ae. albopictus cell lines and tick cell lines
is well known. Cell lines have been attempted
from mosquitoes (C. bitaeniorhynchus ) 3
sand
flies,
ticks and mites.
Efforts are being
made to establish new cell lines from trombiculid mites.
A new method of growing cells on the
microscope slide by using stainless steel
cylinders was tried.
Blood macrophages were
cultured from BCG vaccinated and non-vaccinated donors, to observe the effect of viruses
on this cell line.
Efforts have been made to
develop new cell lines from embryonic kidney.
A new approach for the study of viruses has
been introduced by using chick embryo develop
ing in vitro.
Teratogenic effects of viruses
produced in early chick embryo
(at primitive
streak stage) may provide a new tool for
identification of viruses.
ABSTRACTS
Some Research Projects completed recently
A clinico-epidemic logical study of occlusive
peripheral vascular disease in Delhi
A total of 169 patients of occlusive
peripheral vascular disease forming 0.4 per
cent of the total, annual,
surgical admission
in the Irwin Hospital, were studied. Thromboangitis obliterans
was the most frequent
condition followed by arteriosclerosis obli
terans .
Thromboangitis obliterans was observed
in young males, below 49 years of age (88 per
cent). Gangrene was the commonest symptom and
8
the disease involved both the lower and upper
limbs.
The infrapopliteal pulses were frequently absent.
The cardiovascular system,
electrocardiograms, funduscopic and biochemical examinations were normal,
except for
significantly
raised
blood sugar,
serum
cholesterol and serum triglyceride levels.
98 per cent were smokers,
usually of beedis.
Thromboangitis patients came from the lowest
socio-economic class and were heavy workers,
mainly farmers. Calorie and protein malnutrition was present.
Trauma
and migrating
thrombophlebitis were infrequent.
Arteriosclerosis obliterans occurred in
old people, 60 years of age (72 per cent) or
more.
84 per cent were males and 16 per cent
were females.
Ulcers, with or without gang
rene, were the commonest symptom.
Upper limb
involvement was infrequent.
The femoral and
femoropopliteal pulses were absent
most fre
quently.
Diabetes was present in 36 per cent,
cardiovascular
disease
in
20
per cent,
electrocardiographic changes in 25 per cent
cases.
72 per cent were smokers, usually
cigarettes alone or combined with beedts.
Calorie and protein deficiency was less fre
quent than in patients with thromboangitis and
these patients were largely sedentary workers.
Angiography and histopathological studies
revealed 11 per cent thromboangitis patients
having iliac and femoral artery obstructions.
Blood sugar,
serum cholesterol and serum tri
glyceride levels were significantly raised in
thromboangitis
but were
still higher in
arteriosclerosis.
Thromboangitis obliterans was commonest
in patients from Uttar Pradesh (58 per cent)
while 60 per cent arteriesclerotics and 70 per
cent controls came from Delhi.
Significantly
more than a quarter (28 per cent) of the total
patients were farmers.
Thromboangitis obli
terans occurred mainly in the poorest socio
economic group.
19 per cent patients with embolism, Ray
naud's disease, phlegmasia caerulea dolens,
collagen
diseases
(rheumatoid
arthritis,
scleroderma and systemic lupus erythematosus),
iatrogenic (trauma and drug arteritis), non
specific arteritis,
thoracic outlet syndrome
and questionable tubercular arteritis, in that
order of
frequently,
were also studied,
females prodominated (57 per cent).
R. Nigam and
Colleagues
Departments of Surgery,
Radiology and Pathology
Irwin and G.B. Pant Hospitals
New Delhi.
Effect of hydrogen chloride on lung surfactant
The effect of hydrogen chloride on lung
surfactant of rats was studied.
Healthy male
albino rats were exposed to hydrogen chlo
ride - air mixture for different periods under
dynamic atmosphere at an average concentration
of 440 ppm.
At the end of the exposure,
the
rats were sacrificed and the lung surfactant
activity was assessed by measuring the dynamic
surface tension, drawing air pressure-volume
curves and estimating the phosphdiipid con
tent.
Routine histopathological examination
of the lung tissue was also done.
Exposure of the animals for 5 hours did
not alter the surfactant activity.
Exposure
for brief periods (6 days at 5 hours/day)
indicated a mild decrease in surfactant acti
vity as shown by the increased minimum surface
tension and
reduced phospholipid content.
This was found to be reversible.
Two weeks
exposure showed a definite decrease in sur
factant activity
with incomplete recovery,
indicating that the changes are due to a cumu
lative effect on continued exposure.
With 4
weeks exposure the lung:
surfactant activity
was decreased to a greater extent.
Maximum
and minimum surface tensions were increased,
and phospholipid content was decreased. Static
recoil pressure on deflation of the lung
increased and the retention of air at 0 cm.
of water pressure reduced.
Histopathological
examination revealed . moderate to severe inflammatory reaction of the bronchial wall and
alveolar septa,extensive alveolar hemorrhages,
moderate areas of collapse and in some cases
with mild pulmonary oedema.
Thus,
the lung
surfactant activity was reduced to a mild
degree on exposure to hydrogen chloride for
brief periods and to a greater extent due to
the cumulative effect of prolonged exposure
producing alveolar collapse and oedema.
B. Krishnan
Department of Biophysics
Jawaharlal Institute of
Postgraduate Medical
Education § Research
Pondicherry.
High tibial osteotomy in
tis of knee
degenerative arthri
Evaluation of the role of high tibial
osteotomy in the treatment of osteoarthritis
knee was undertaken in 30 patients.
It was
observed that age had no bearing on the re
sults though stage of the disease had. Results
were good in 19 cases (63.33 per cent),
fair
in 7(23.33 per cent) and poor in 4(13.33 per
cent) .
Relief of pain (75 to 100 per cent)
was seen in 22 cases (73.33 per cent). Results
were better in cases with varus deformity and
medial compartment involvement.
Correction
and maintenance of the femoro tibial angle had
a definite bearing on the results of osteo
tomy. Best results were achieved if the angle
was more than 10° valgus.
9
It was felt that this is a simple pro
cedure, not requiring prolonged postoperative
immobilisation thereby reducing the load on
hospital beds; with early union of osteotomy,
without any danger of restriction of movements
and with good relief of pain.
It was observed that in all the cases
(21) studied absorption and consolidation of
the bone graft occurred. This was seen earlier
in cancellous bone than in cortical bone.
Immunological response
due to
homogenous
grafting was not observed in any of the cases.
Observation of ether preserved homogenous bone
grafts in human subjects
The sterility of the graft was maintained
by ether preservation, which did not alter
qualitatively and quantitatively the alkaline
and acid phosphatase contents of the bone in
24 hours; and histological pattern of the
bone.
Ether thus seems to be a suitable pre
servative for homogenous bone grafts and could
be tried for establishing bone banks.
With a view to assess whether ether pre
served homogenous bone grafts could act as
substitutes for autogenous grafts and deep
freeze grafts, this study was undertaken using
ether preserved bone grafts in human beings,
following various orthopaedic operations.
B.K. Singh
R.R. Ganguly
Department of Orthopaedic Surgery
Darbhanga Medical College
and Hospital
Darbhanga.
A.K. Gupta
P.S. Maini
Department of Orthopaedics
Medical College
Rohtak.
IC MR
NEWS
The Virus Research Centre, Pune, has been
renamed as the National Institute on Virology
on 1st August, 1978, in recognition of its ex
panding activities in the field of virology.
The
Scientific Advisory Committee of the
Institute met in Pune on 1st and 2nd August,
1978.
Dr. S.C. Pal, Director, Cholera Research
Centre,
Calcutta, participated in the Nobel
Symposium on •Cholera and related diarrhoeas molecular aspects of a global health problem1
held at Stockholm, Sweden,
from 6th to 11th
August, 1978.
Dr. S.G. Srikantia,
Director, National
Institute of Nutrition, Hyderabad,participated
in the WHO consultative meeting on ‘Development
of methodologies and guidelines for*
training of PHC workers in nutrition1 held at
Geneva between 7th and 11th August, 1978.
Dr. C. Gopalan,as President of the Inter
national Union of Nutritional Sciences, parti
cipated in the XI International Congress of
Nutrition held at Rio de Janeiro, Brazil, from
27th August to 1st September, 1978.
Dr. P.K. Rajagopalan,
Deputy Director,
Vector Control Research Centre,
Pondicherry
participated in the meeting of the Technical
Review Group III of the WHO Special Programme
for Research and Training in Tropical Dis
eases, held at Geneva between 28th August and
1st September, 1978.
PROGRAMME OF SCIENTIFIC LECTURES AT THE
INDIAN COUNCIL OF MEDICAL RESEARCH, NEW DELHI
Date
Topic
Speaker
8.9.1978
Importance of enteroviruses in India
(Dr. J.B. Shrivastav Memorial Lecture)
Dr. T. Jacob John,
Vellore.
22.9.1978.
Immune deficit in lepromatous leprosy and its
correction
Dr. Kunal Saha,
New Delhi.
10
ICMR AIDED SYMPOSIA/SEMINARS/WORKSHOPS
A laboratory teaching workshop in Physio
logy was organised by the Department
of
Physiology,
All India Institute of Medical
Sciences, New Delhi, between 17th and 29th
July, 1978.
A seminar on *Care of a worker* was
organised by the Indian Association of Occu
pational Health at Baroda on 22nd and 23rd
July, 1978.
A symposium on
‘Clinical
pharmacoki
netics*
is being organised during the V
Southern Regional Conference of the Indian
Pharmacological Society at Pondicherry on 9th
and 10th September, 1978.
Symposia on ‘Agricultural and food bio
chemistry*,
’Nutritional and clinical bio
chemistry*
and ’Separation methods in bio
chemistry’ are being organised by the Society
of Biological Chemists of India at Delhi
between 3rd and Sth October, 1978.
A symposium on ’Surgery of the internal
auditory meatus’ is being organised at the All
India Institute of Medical Sciences,New Delhi,
on Sth and 6th November, 1978.
A symposium on ’Brain pituitary adreno
cortical inter-relationship’
is being held at
the Institute of Medical Sciences, Varanasi,
between 6th and Sth November, 1978.
A workshop on ’New techniques in neurosciences with special reference to the neuro
physiology of higher nervous activity’ is
being organised at the National Institute of
Mental Health and Neurosciences, Bangalore,
between 6th and 17th November, 1978.
A symposium on ’Recent developments in
applied analytical chemistry*
is being orga
nised by the Institution of Chemists (India)
at the Medical College, Calcutta, on 11th and
12th November, 1978.
A
care’
seminar
is being
on ’Engineering and health
organised by the Instrument
Society of India at Mysore, in November, 1978.
A symposium on ’Population structure and
human variation’ is being organised during the
International Congress of Anthropological and
Ethnological Sciences at Bombay between 19th
and 21st December, 1978.
Symposia on ‘Isoenzymes in clinical bio
chemistry’ and ’Hormonal analysis in clinical
biochemistry’ are being organised during the
V Annual Conference of the Association of
Clinical Biochemists of India at the Seth G.S.
Medical College and K.E.M. Hospital, Bombay,
between 27th and 30th December, 1978.
A symposium on ’Problems in drug therapy
in National health programmes’ is being orga
nised during the XI Annual Conference of the
Indian Pharmacological Society at the All
India Institute of Medical Sciences,New Delhi,
between 29th and 31st December, 1978.
A symposium on
’Indigenous drugs in
clinical medicine: Problems and possibilities’
is being organised during the XXIV Annual Con
ference of the Association of Physiologists
and Pharmacologists of India at the RNT Medi
cal College, Udaipur, in December, 1978.
A symposium on
’Host-parasite inter
action’ is being organised at the Department
of Zoology, University of Madras, Madras, in
December, 1978.
Symposia on ’Reproductive biology’ and
’Nutrition’ are being organised during the VII
International Congress of the Primatological
Society at the Indian Institute of Science,
Bangalore, between 1st and Sth January, 1979.
A symposium on ’Mechanisms of aggressive
behaviour*
is being organised during the
Winter School on Neurophysiological and Neuro
chemical Correlates of Behaviour at the All
India Institute of Medical Sciences,New Delhi,
between 19th February and 3rd March, 1979.
11
COUNCIL'S TRAINING PROGRAMMES FOR 1978
Lebeess
Haematology:
At the Central Jalma Institute for
rosy, Agra.
Training course in Leprosy
officers:
(December, 1978).
At the
Bombay.
Lep
for medical
(1)
Diploma course in Virology
versity:
(July, 1978).
(2) *Training course for
period of one year.
(3)
Short-term course in Animal Cell
Culture:
(June, 1978).
for a
At the Institute for
duction, Bombay.
Research, in
Centre,
(2)
Training course in Blood Banking Methodo
logy for Technicians: (May 2 to 14,1978).
Occupational Health:
At the National Institute of Occupational
Health, Ahmedabad.
Tissue
Reproductive Biology:
Reference
A certificate course in Blood Banking
Technology and Basic Haematology
for
medical officers:
(February 1,
1978 to
March 31, 1978).
of Poona Uni
Technicians
Group
(1)
Virology:
At the Virus Research Centre, Poona.
Blood
(1)
ICMR/WHO training course
Toxicology: (March 1978).
(2)
Refresher course in
for E.S.I. medical
1978) .
(3)
Training course on Chemical Hazards in
Chemical Industry organised jointly by
the NIOH and the National Safety Council:
(December 1978) .
in Industrial
Repro
(1)
Training course in Reproductive Biology:
(January, 1978 to March, 1978).
(2)
Radioimmunoassay of Protein and Steroid
Hormones: (January, 1978 to March, 1978).
(3)
Training course in Genetics:
1978 to February 15, 1978) .
Occupational Health
staff:
(September
(January 16,
Clinical Pharmacology:
(4)
Training course in
Basic Immunology:
(February, 1978 to March, 1978).
At the Department of Pharmacology,
G.S. Medical College, Bombay.
(5)
Training course in Contraceptive Research
and Reproductive Endocrinology:
(March,
1978).
Training course in Clinical Pharmacology:
(January 15, 1978 to February 28, 1978).
Biostatistics:
Nutrition and Endocrinology;
At the National
Hyderabad.
Seth
At the National' Institute of Nutrition,
Hyderabad.
Institute of Nutrition,
(1)
M.Sc. (Applied Nutrition) course:
1978 to February, 1979) .
(2)
Annual certificate course in Endocrino
logical Techniques and their Application:
(October, 1978 to November, 1978).
Annual training course in Biostatistics
for Statisticians:
(November, 1978).
(June,
Laboratory Animal Technology:
(3)
12
Annual certificate course in Nutrition:
(December, 1978 to February, 1979).
(1)
At the Virus Research Centre, Poona.
Short-term course in Breeding and Mainte
nance of
Laboratory Mouse
Colonies:
(March, 1978).
(2)
At
the National Institute of Nutrition,
Hyderabad.
Clinical Chemistry and Medical Instrumenta
Laboratory Animal Technicians Training
Course:
(August,1978 to September, 1978) .
At the National Institute of Nutrition,
Hyderabad: (July 10 to 16, 1978).
COURSES FOR TALENT SEARCA SCHEME FELLOWS
tion:
Use of Radioisotopes in Medicine:
At the National Institute of Nutrition,
Hyderabad: (July 24 to August 2, 1978).
Haematological Techniques:
(1)
(2)
At the Department of Pathology, Christian
Medical College, Vellore:
(February 28,
1978 to March 10, 1978).
At the Department of Haematology,
*
Postgraduate Institute of Medical Education
and Research,Chandigarh: (September, 1978) .
Cell-Biology and Cyto-Genetics:
At
the Indian Institute of Science,
Bangalore: (January 1 to 10, 1979).
*Principles and Methods in Medical Statistics:
At the Tuberculosis Chemotherapy Centre,
Madras.
Clinical Pharmacology:
*Methods used in Diagnostic Virology:
At the Department of Pharmacology,
Post
graduate Institute of Medical Education §
Research,
Chandigarh:
(March 6 to 15,
1978).
SOME
ICMR
At the Virus Research Centre, Poona.
*Dates to be announced later.
PUBLICATIONS
TECHNICAL REPORTS
Price
Rs.
Research in Medical Education (1970)
4.00
Research in Health Practices (1970)
4.00
Acute Encephalopathy Syndrome in Children (1970)
4.00
Exfoliative Cytology (1971)
4,00
Symposium on Nutrition and Heart Disease
(1971)
4.00
Developments in Industrial Psychology in India (1971)
4.00
Proceedings of Seminar on Immunity and Immunoprophylaxis in Cholera (1971)
4.00
Diagnosis and Treatment of Pesticide Poisoning (1971)
4.00
Symposium on Clinical Evaluation of New Drugs (1972)
4.00
Techniques in Blood Group Serology (1972)
4.00
Laboratory Methods in Investigation of Thyroid Diseases (1972)
4.00
Proceedings of the First Annual Meeting of the Indian Academy of Cytologists
(1972)
4.00
Review of Work Done in Rural Water Supply in India (1972)
Genetics and our Health (1972)
4.00
4.00
13
Price
Rs.
All India Conference on Research in Reproduction and Fertility (1973).
4.00
A Study of Morbidity Patterns and Standard of Medical Care in the
Rural Centres of West Bengal (1973)
4.00
Proceedings of the First All-India Workshop Conference on Stroke (1973)
4.00
Seminar on Infectious Hepatitis and Study Group on Australia Antigen (1973)
4.00
Atlas of Histopathology of Liver: Hepatitis and Cirrhosis (1974)
10.00
Studies on Pre-school Children (1974) Reprinted 1977
3.50
Studies on Weaning and Supplementary Foods (1974) Reprinted 1977
4.00
Medicinal Plants of India - Vol.I (1976)
30 00
Alternative Approaches to Health Care (1977)
Gratis
Research in Malaria - An Outline (1977)
Gratis
These publications are available on prepayment of cost by cheque or postal order
drawn in favour of the Director-General,
Indian Council of Medical Research, New
Delhi. Money orders are not acceptable. All correspondence in this regard should be
addressed to the Chief,
Division of Publication and Information,
Indian Council of
Medical Research, Post Box 4508, Ansari Nagar, New Delhi-110016.
EDITORIAL BOARD
Chairman
Dr. C. Gopalan
Director-General
Chief, Publications
Dr. G.V. Satyavati
Members
Dr. S.V. Apte
Dr. N.D. Datta Banik
Dr. Usha K. Luthra
Dr. V.N. Rao
Dr. A. D. Taskar
Dr. Sabita Tejuja
Editor
Dr. N. Medappa
Printed and Published by Shri Prem Chand for the Indian Council of Medical
Research, New Delhi at the ICMR Offset Press, New Delhi.
R.N. 21813/71
Position: 1894 (15 views)