76
this organism was not isolated from the urine of any oi
them before or after instrumentation. With the exception
of 1 patient with renal papillary necrosis, in whom the
organism persisted in the urine for 15 months, it seemed tc
be almost non-pathogenic. Blood:-cultures from this
patient, both after urethral instrumentation and at other
times, were always negative. We attributed the high
incidence of bactersemia with this organism after trans-
urethral resection (11 cases out of 18) to the combined
influence of more vigorous bladder irrigation and an
extensive raw area as compared with the other procedures.
Blood-stream infection induced mechanically by the
passage of an instrument along the urethra is an accepted
hazard. We were not aware that direct blood-stream
invasion from contaminated bladder-irrigating solution
could be so frequent. Contamination of theatre " sterile "
water by a more dangerous organism could represent a
major hazard.
An attempt has been made to overcome blood-
stream invasion from this source by the use of distilled
water dispensed in one-litre flasks and sterilised by
autoclaving.
Preliminary Communication
CULTIVATION OF VIRUSES FROM A HIGH
PROPORTION OF PATIENTS WITH COLDS
WITH present methods of tissue culture and testing, it
is usually possible to cultivate a virus from about a quarter
to a third of adult patients with common colds. Organ
cultures of human foetal tracheal or nasal epithelium have
been shown to support the growth of representative strains
of all known respiratory viruses.l2 These cultures have
also been used to cultivate some apparently " new "
viruses-namely (a) 2 rhinoviruses which will produce
a cytopathic effect in human diploid-cell strains only after
passage in organ culture 3(b) a rhinovirus which will not
grow at all in such strains 4; and (c) an ether-labile virus
apparently unrelated to any of the known ether-labile
viruses of the human respiratory tract.3
Using organ cultures we have recently attempted to
cultivate viruses from a series of patients with colds and
similar acute respiratory illnesses.
METHOD
The
specimens consisted of nasal washings collected at
irregular intervals between 1961 and 1964, and more regularly
since then from 21 adults and 2 children shortly after the onset
of illness. The patients were laboratory staff and their families
and members of the general public in whom spontaneous colds
developed while they were staying at the Unit. Specimens
were not collected when it seemed likely on epidemiological
grounds that the infection had been caused by a virus that had
already been collected. We also tested four pools of washings
taken from volunteers infected with strains of virus which we
had repeatedly failed to cultivate. The washings were made
with isotonic phosphate-buffered saline, pH 7-1, and were
mixed with an equal volume of nutrient broth and stored at
-
70°C until tested.
"
All specimens were tested in standard " tissue cultures of
monkey kidneys, HeLa cells, human diploid cell strains, and
human foetal kidney.5 The specimens were also inoculated into
orsan cultures 1; medium was collected daily from these
1. Hoorn, B., Tyrrell, D. A. J. Br. J. exp. Path. 1965, 46, 109.
2. Tyrrell, D. A. J., Hoorn, B. ibid. p. 514.
3. Tyrrell, D. A. J., Bynoe, M. L. Br. med. J. 1965, i, 1467.
4. Hoorn, B., Tyrrell, D. A. J. Arch. Virusforsch. (In the press).
5. Medical Research Council Working Party on Acute Respiratory Virus
Infections. Br. med. J. 1965, ii, 319.
Summary
In 100 men undergoing urological surgery, urine and
blood cultures were taken before and after urethral
instrumentation. Twenty-two positive blood-cultures
wereobtained, eight of which grew common pathogens
and fourteen an apparently non-pathogenic organism
which had been an intermittent contaminant of the theatre
"
sterile " water-supply. Only five of the positive blood-
cultures grew the same organism as had previously been
present in the urine. 11patients had a fever after the
procedure but only 1 showed the clinical picture of
septicaemia. Antibiotics were given to 4 of these patients
before instrumentation and to 10 afterwards. It appears
that bacteria can enter the blood-stream as a result of
bladder irrigation, particularly in transurethral resection.
We wish to thank the chairman of the Repatriation Commission
for permission to publish this work.
Requests for reprints should be addressed to Dr. P. M. Last,
Repatriation General Hospital, Daws Road, Springbank, South
Australia.
REFERENCES
Bulkley, G. J., O’Connor, V. J., Sokol, J. K. (1954) J. Urol. 72, 1205.
Creevy, C. D., Feeney, M. J. (1954) ibid. 71, 615.
Mitchell, J. P., Slade, N., Linton, K. B. (1962) ibid. 34, 454.
between the second and tenth day and stored with broth as for
nasal washings. The fluids were then combined, and the pool
was retested in standard cultures; if negative, it was inoculated
intranasally into 6 or more volunteers. If 1 or more volunteers
got colds the specimen was taken to contain a virus. In most
cases two passages were made in organ cultures.
RESULTS
The results obtained so far are outlined in the accom-
panying table. The rate of virus isolation in standard
cultures was about that expected, and the viruses were of
various types with rhinoviruses predominating. The
rhinoviruses were identified by their biological properties,
but were not serotyped. All the viruses recognised by
direct inoculation of standard cultures were also recognised
by testing the media of organ cultures inoculated with the
same washing. However, additional rhinoviruses were
cultivated in organ cultures and were then readily recog-
nised by their effect on diploid cells, although the original
specimens were negative when tested in these cells. Some
of these viruses were difficult to propagate and identify,
because they grew to low titres in human diploid fibro-
blast cells. In addition, six further cold-producing agents
TESTS ON 33 NASAL WASHINGS FROM PATIENTS WITH COMMON COLDS
AND SIMILAR ACUTE UPPER RESPIRATORY DISEASES
*
Numerator equals number of cultures producing colds in 1 or more
volunteers and denominator equals number of specimens tested.
+ Ciliary activity was reduced in organ cultures inoculated with 5 of these
rhinoviruses.
 77

were detected because organ culture fluids caused colds in
volunteers. The tests are incomplete, but no reduction in
ciliary activity was noted in cultures in which these agents
were growing. Altogether, therefore, twenty-five viruses
or other agents were recovered from 33 specimens (75%)
and nineteen of these (57%) were recognised in the
laboratory. Of the 8 specimens from which no virus was
isolated, 6 were still available and were inoculated into
volunteers. 3 of these produced colds; hence it is clear that
there are a few cold-producing agents which cannot at
present be propagated in organ cultures. We conclude,
nevertheless, that these organ cultures were an efficient
method of propagating viruses from patients’ colds. The
combination of organ culture with diploid fibroblast cell
cultures revealed the presence of nine more rhinoviruses
than could be found by standard techniques. But we
obviously need to devise simple methods of detecting some
of the other new agents in organ culture medium and then
to characterise them. Some at least may resemble the
B814 virus described earlier.3
DISCUSSION
Against possibility that the viruses might originate
the
from the organ cultures is the finding that a cold developed
in only 1 out of 113 volunteers who were given fluids from
uninoculated cultures made from embryos used in these
experiments and that no viruses were isolated by inocu-
lating the same fluids into " standard " tissue cultures.
Washings collected from 11 persons who had recovered
from their colds were also tested; a rhinovirus was isolated
in organ culture from 1 woman who had carried a similar
virus two weeks before, when she had a cold; and one
culture fluid caused a cold in a volunteer. Thus there were
two apparent isolations from 11 specimens; this is a
significantly lower rate than the rate obtained by all
methods from all patients with colds (p < 0’01) and the
rate from the 23 patients from whom viruses were not
isolated by standard methods (p < 0-05). These figures
and the typical clinical picture produced in volunteers
show that the agents isolated are the genuine cause of the
’,-
colds.
SUMMARY
A virus or uncharacterised cold-producing agent was
cultivated from 25 of 33 nasal washings by inoculating
them into organ cultures of human-embryo nasal or
tracheal epithelium.
D. A. J. TYRRELL
Medical Research Council M.D. Shelf., F.R.C.P.
and Ministry of Health M. L. BYNOE
Common Cold Research Unit,
Salisbury, Wilts M.B. Lond., D.T.M. & H.

New Inventions intestines may be out-

lined. Holman and
Karnicki5 seem to
AN AID TO INTRAUTERINE TRANSFUSION
dispense with this
OF THE F&OElig;TUS FOR aid. Risk to the
ERYTHROBLASTOSIS F&OElig;TALIS foetus is inherent in
INTRAPERITONEAL blood-transfusion of the foetus in utero in this technique if
severe erythroblastosis foetalis is rapidly becoming a standard localising aids to
make insertion of the
technique in selected cases. This development has been made
needle as precise as
possible by the work of Bevis,l Walker,2and Liley3 with
spectrophotometric analysis of liquor amnii. Two distinct possible are not used.
methods of intrauterine transfusion have been reported. The earlier the gesta-
tion the greater the
Liley’s is the one generally used and involves the placing of a
needle or catheter in the foetal peritoneal cavity. When the difficulty in defining
foetus was too small for this technique Adamsons et al. opened the position of the
the uterus and implanted a T-tube catheter in the foetal foetus accurately. In
the obese patient also
peritoneal cavity.
The Liley procedure involves the injection of contrast the position of the
medium into the amniotic cavity to outline the foetus. The fcetus may be difficult
to determine.
baby may also swallow some of the medium and the foetal
1. Bevis, D. C. A. Lancet, 1952, i, 395. Fig. 2-Grid superimposed on antero- METHOD
2. Walker, A. H. C. Br. med. J. 1957, ii, 376. posterior film. Lead markers as arbi-
3. Liley, A. W. Am. J. Obstet. Gynec. 1961, 82, 1359. trary guides. Hypaque ’ in foetal During the last
4. Adamsons, K., Jr., Freda, V. J., Plentl, A. Personal communication,
1965
intestinal tract. eight intrauterine
transfusions per-
formed at the Yale-
New Haven Medical
Center, we have used
a grid as well as the
intra-amniotic injec-
tion of contrast me-
dium. This grid is
composed of fine
stainless steel wires
1/2 in. apart mounted
on a frame (fig. 1).
The anteroposterior
X-ray picture of the
maternal abdomen
shows the grid. The
square on the X-ray
Fig. 3-Lateral marker superimposed on that
lateral film. Marker is placed adjacent permits the best
to film cassette and thus no correction
access to the foetal
factors are needed. peritoneal cavity is
Fig. 1-Grid in position for anteroposterior film. 5. Holman, C. A., Karnicki, J. Br. med. J. 1964, ii, 594.