ORNITHOSIS ISOLATION OF THE VIRUS FROM A CASE

696 it is clear that the amount of lactose in the diet will determine the severity of the lesion, thus emphasising the truth that what is inherited is not a character but merely a predisposition, the actual degree of" realisation of which depends on environmental factors. Brolin and Krakau (1949) have shown that the occurrence of this type of cataract in rats is directly proportional to the percentage of galactose in the diet. It is, for example, probable that, in a case of galactose intolerance, if the infant receives no lactose at any time it may develop

parallel,

ORNITHOSIS ISOLATION OF THE VIRUS FROM A CASE

P. R. DUNCAN Mane., M.R.C.P.

M.B., B.Sc.

SENIOR MEDICAL REGISTRAR

we

SUMMARY case

MEDICAL REGISTRAR

J. O’H. TOBIN Oxfd, Dip. Bact.

B.M.

have travelled a long way from the position adopted by Gates (1929) when he said of congenital cataract, probably quoting Macklin (1927), that " there are many types all with strong evidence of inheritance." We now know that some cases are due to the action of the rubella virus during pregnancy (cases in which the genetic element is presumably unimportant) : 5 such cases in the Fountain Hospital with a population of 650 are attributed to this factor. We have further the of intolerance to example galactose suggest that in the remaining cases environmental factors may be just as important as genetic. A static arrangement of the facts to fit preconceived genetic patterns in terms of " 2 dominant and 3 recessive factors " leads to pessimism and therapeutic nihilism in the field of mental deficiency. A chemical examination and reformulation of the data may be of great help in elucidating some of the problems in this field, may make therapy, if started early, worth while, and will at least render the practitioner better able to advise the parents of a defective child who consult him about the outcome of future pregnancies. Duke-Elder’s (1943) statement still holds good that "the setiology of these opacities depends upon some disturbance in the development of the lens, but what the actual disturbance may be, or the precise method of its action, is a matter of considerable doubt in most cases." It seems, however, reasonable to hope that, with a proper concentration of research on this problem, the nature and mode of action of the noxious factors may be understood in a considerable proportion of cases.

A

M.B., B.Sc. Manc., M.R.C.P.

MANCHESTER ROYAL INFIRMARY

normally. Thus

A. E. THOMAS

of

congenital

imbecility (Langan

i.Q.

familial bilateral cataract with

49) is described. In three genera-

tions 4 persons were affected by cataract, mental deficiency, and a 6th by epilepsy. A tolerance test gave negative results.

5th by galactose-

a

due to Dr. D. A. Pond for the report on the to Dr. R. R. Race for the bloodgroups ; to Dr. L. I. Woolf for the biochemical reports and the interesting suggestion that galactose intolerance might be a causal factor ; to Dr. W. W. Kay for the serological examinations ; and to Mr. L. H. Savin for the report on the eyes. Our thanks

are

electro-encephalogram;

REFERENCES

LECTURER

p. 112.

Harman, N. B. (1909) Trans. ophthal. Soc. U.K. 29, 296. Knapp, F. N. (1926) Amer. J. Ophthal. 9, 683. Lowe, R. F. (1949) Brit. J. Ophthal. 33, 131. Macklin, M. T. (1927) Canad. med. Ass. J. 17, 940. Manson, J. S. (1928) Observations on Human Heredity. London ; p. 4. Penrose, L. S. (1949) Biology of Mental Detect. London ; p. 94. Saebø, J. (1949) Brit. J. Ophthal. 33, 601. Wilson, S. A. K. (1940) Neurology. London ; p. 104. Worth, C. (1930) May and Worth’s Diseases of the Eye. London ; p. 239.

BACTERIOLOGY IN THE MANCHESTER

UNIVERSITY

OF

THE virus of ornithosis, a member of the psittacosis lymphogranuloma-venereum group, closely resembles that of psittacosis but can be distinguished by difference!-. in its infectivity for animals. Psittacosis is a disease of psittacine birds (parrots and budgerigars) which is

communicable to man. The term ornithosis is used for similar infections occurring among a wide range of non-psittacine birds, including fulmars, domestic fowls, pigeons, and ducks. The ecology of psittacosis and ornithosis has been fully reviewed by Meyer (1942). The virus of psittacosis has been isolated from human cases of respiratory infection (Bedson et al. 1930, Troup et al. 1939), but isolation of the virus of ornithosis has not yet been reported in this country. Recovery of the ornithosis virus from the throat-washings of a poultry dealer with atypical pneumonia and serological evidence of recent infection with this virus is therefore described here. CASE-RECORD

A male poultry dealer, aged 42, was admitted to the Manchester Royal Infirmary on Nov. 7, 1950, under the care of Dr. C. S. D. Don, with frontal headache, cough, and pain across the back of the lower chest. His illness had started suddenly four days previously with shivering and sweating. He had been treated by his doctor with a sulphonamide for three days without effect. On admission he looked ill and was perspiring freely but mentally alert. His temperature -was 100-6°F, with pulse-rate 95 and respirations 28 -per min. Numerous coarse crepitations were audible over the right, and to a lesser degree the left, lower lobes. No other abnormality was discovered on physical examination. A radiograph of the chest was normal. was

Treatment and Progress.-Next day his condit,ion way unchanged. As he complained of persistent headache, his cerebrospinal fluid was examined and found to be normal. Specimens of blood and throat washings were sent for virus-

isolation, and

treatment with

units six-

penicillin 200,000

hourly was started. Two days after admission he was apyrexial and feeling much better. Penicillin was discontinued eight days after admission, and the remainder of his stay in hospital was without incident. He returned home on Nov. 20, 1950. A further chest radiograph on that date was

Alger, L. G. (1945) Amer. J. Ophthal. 28, 730. Bell, J. (1932) Treasury of Human Inheritance. London ; vol. II, part v, p. 527. Brolin, S. E., Krakau, C. E. T. (1949) Acta ophthal., Kbh. 27, 291. Danforth, C. H. (1914) Amer. J. Ophthal. 31, 161. Duke-Elder, W. S. (1934) Chances of Morbid Inheritance. Edited by C. P. Blacker. London ; p. 172. (1943) Textbook of Ophthalmology. London; p. 1356. Ford, E. B. (1948) Genetics for Medical Students. London ; p. 150. Gates, R. R. (1929) Heredity in Man. London ; p. 97. Goldstein, E. O., Ennis, J. E. (1948) J. Pedlatr. 33, 147. Gregg, N. M. (1941) Trans. Ophthal. Soc. Aust. 3, 35. Gruneberg, H. (1947) Animal Genetics and Medicine. London;

IN

normal.

Other investigations.-On Nov. 7, 1950, a blood-count showed red cells 5,300,000 per c.mm., Hb 15-4 g. per 100 ml., white cells 7700 per c.mm. (polymorphs 78%, lymphocytes 17%). Cold agglutinin and Paul-Bunnell tests were both negative on Nov. 8. Normal upper-respiratory-tract organisms were grown from the sputum on Nov. 9, and no tubercle bacilli were seen on microscopy, A Frei test done on Nov. 27 was

negative.

’

’

VIRUS STUDIES

,

As the

patient lived near an area in which lymphocytic choriomeningitis is endemic (Duncan et al. 1951), the virus of this disease was considered- to be- a. possible cause of his atypical pneumonia. 5 ml. of throat washings and 2 ml. of blood were each injected intraperitoneally into two pairs of guineapigs. After five days one animal from each pair was pyrexial. These ’

’

,

"

two

were

killed,

and

a

injected intracerebrally

1 in 20 emulsion of their spleens was into two batches ·of mice.

697

t111 the mice inoculated

with the emulsion from the

guinea-

with blood died of Salmonella pig which had been injected other batch became ill with The MMMt; inaction. ruffled fur, tremors, and slight conjunctival exudato, and were

moribund by the fifth day. Smears from the brains of these mice contained numerous pus cells but no organisms ; aerobic and anaerobic cultures were sterile. Intracerebral passage of brain from these animals produced the same clinical features in a second batch of mice. Smears from the brains of these mice showed numerous pus cells and elementary bodies morphologically identical with those of psittacosis. A 1 in 10 suspension of brain from these mice was infective when diluted 10-4 and injected intracerebrally into other mice. Intraperitoneal injection of 0-5 ml. of the suspension was either without visible effect, or the mice developed ascites from which they recovered. After ten days no virus could be isolated from their spleens. These findings indicated that the infective agent

was

the virus of ornithosis rather than that of psittacosis, which is pathogenic to mice by both the intracerebral and intraperitoneal routes (Meyer et al. 1942). In the complement-fixation test allantoic fluid containing the virus isolated from the patient was used as antigen because the virus grew well in the allantoic cavity of the developing chick embryo. The results of this test were :

Our thanks

due to Dr. C. S. D. Don for permission to S. P. Bedson, F.7,.s., Mr. J. Norris, superintendent of Manchester Town Hall, for his department’s help in obtaining pigeons for examination ; and to Prof. H. B. Maitland for helpful criticism and advice. are

publish details of the patient ; to Prof. for help in confirming the diagnosis ; to

REFERENCES

Bedson, S. P., Western, G. T., Simpson, S. L. (1930) Lancet, i, 235. Duncan, P. R., Thomas, A. E., Tobin, J. O’H. (1951) Ibid, i, 956. Meyer, K. F. (1942) Medicine, Baltimore, 21, 175. Eddie, B., Yanamura, H. Y. (1942) Proc. Soc. exp. Biol., N.Y. 49, 609. Tasker, J. R. (1949) Brit. med. J. ii, 362. Troup, A. G., Adam, R., Bedson, S. P. (1939) Ibid, i, 51. Wolins, W. (1948) Amer. J. med. Sci. 216, 551. —

THE ESTIMATION OF FORMALDEHYDOGENIC STEROIDS A. A. HENLY

MARJORIE POTTER

Ph.D. Lond. ROCHE FELLOW IN BIOCHEMISTRY EMPIRE RHEUMATISM

From the Maclean

RESEARCH ASSISTANT COUNCIL

Laboratory, Hospital of St.

John

and

St. Elizabeth, London

by Hollander et a]. (1951) for estimating formaldehydogenic steroids the formaldehyde is determined by isothermal distillation directly- into the chromotropic acid reagent using a Conway micro-diffusion unit. After distillation is complete, the chromotropic acid-formaldehyde mixture is transferred to a test-tube IN the method used

Professor Bedson kindly confirmed the presence of elementary bodies of the psittacosis group in the mousebrain smears. He also found that the serum obtained on the eleventh day fixed complement with psittacosis antigen. Fixation was complete at a dilution of 1 in 16 and almost complete at 1 in 32. Complement-fixation tests with the serum of two members of the patient’s family and of the assistant in his shop were

negative. The source of this patient’s infection was not determined. However, the virus of ornithosis was isolated from one of six pigeons trapped in the Manchester area. DISCUSSION

Ornithosis

was

diagnosed

in this

case

for the

following

(1) the virus was pathogenic to mice by the intracerebral route only, while that of psittacosis is also pathogenic by the intraperitoneal route ; (2) mousereasons :

adapted virus grew well in the allantoic cavity of the developing chick embryo without the preliminary yolkaac passage required for the virus of psittacosis ; and (3) the patient handled large numbers of pigeons, fowls, ducks, and turkeys in his poultry shop. Although a budgerigar was kept in his home it had uot been sick ; nor had any member of the patient’s family had an illness suggestive of atypical pneumonia. No

conclusion

penicillin

on

the

can

be drawn about the effect of of this patient’s illness ; for the twelve hours before the tempera,

course

treatment began only ture fell and clinical

improvement started. Wolins (1948} reported satisfactory response to treatment with penicillin 400,000 units daily in 4 of 6 patients with ornithosis ; the same dose of penicillin was without effect in the remaining 2. Tasker (1949) has recorded a case of psittacosis in which there was a dramatic response to,treatment with penicillin 2,000,000 units daily. a

SUMMARY

Ornithosis infection in a poulterer is described. The methods used for isolation and identification of the virus are recorded. The serological evidence of infection is given.

cuvette and heated at 100°C for 30 minutes, and the colour is measured at 570 m!L in the usual way. This method would be easier if the colour development could be -carried out in the Conway unit. This can be done by allowing the distillation to take place at 50°C. We have found the following procedure to be the most

satisfactory. 1 ml. of

formaldehyde-containing

solution

(0-4-4 {jLg. per

ml., preferably in 5% acetic acid) is pipetted into the inner chamber of a standard Conway unit (Gallenkamp no. 1), the outer chamber of which contains 2 ml. of 0;2% chromotropic acid in 15M sulphuric acid. The unit is sealed and placed in an oven at 50°C overnight (17 houra). Appropriate blanks are also set up. Next morning the extinctions are measured at 570 met (’ Unicam D.G.’ spectrophotometer, 1 cm. cells).

Under these conditions recovery of formaldehyde is quantitative (>95%), the colours formed being- equal to those obtained by heating mixtures of 1 ml. of formaldehyde solution and 2 ml. of chromotropic reagent at100°C for 30 minutes. 1-5 {jLg. of HCHO gives 1 lÌ extinction of about 0.26, the relationship between extinction and concentration being linear over -the range cited above. The specific colour is very stable; and after reaching its maximum it appearsto be unaffected by further heating for several hours. The recovery of formaldehyde from perio"date-oxidised - solotions of cortisone is also quantitative. The equivalent of 5-50 !Lg. of cortisone can be estimated satisfactorily, 25 Ug. giving an extinction of 0-35. Other steroids are "" being studied; -

-

This method has been used for the estimation of formaldehydogenic steroids (F.s.) in urinary extracts of the steroid conjugates with prepared after P-glucuronidase. The F.s. excretion of several rheumatoid arthritic women, aged 40-50, was found to be 6.513-0 mg. per 24 hours, these results being some 10-20 times greater than those obtained after hydrolysis at pHlfor 18 hours, as in older methods. In preliminary experiments, a considerable increase in F.s. excretion was noted after administration of A.C.T.H. by intramuscular or intravenous infusion during 8 hours. Thus; in two patients who received 20’i.tr. intravenously, the

hydrplysis