S. Mallannah
1907 The Lancet  
IT is a well-known clinical fact that the cases of plague which generally recover are those in which the glandular reaction is most marked, whilst the most serious cases are those in which the glandular reaction is hardly observable. It is also observed in the laboratory that marked pathological changes occur in lymphatic glands, liver, and spleen, especially in those animals in which the disease from some cause or other runs a chronic course. I found, for example, marked necrotic changes in
more » ... :e organs in animals partially protected with Haffkine's fluid, and afterwards dying from plague when subsequently infected with the bacillus pestis. These facts induced me to consider that the organs on which the plague poison exercises its concentrated effect may probably be the centres where the antibodies are manufactured in great quantity in case of recovery. In this way I was led to consider whether it was possible to use some form of glandular extract from immunised animals as a curative agent in plague. In order to test the practicability of this theory I carried out some experiments in Hyderabad in the year 1900 and found that the glands of the immunised animals, such as the liver, the spleen, and the buboes when finely minced aseptically, and treated with absolute alcohol for two weeks or more and then allowed to dry at the room-temperature, could be easily converted into a fine powder. The powder thus obtained when well rubbed down in a sterile mortar with a sterile normal salt solution yielded on filtration a clear straw-coloured liquid which seemed to possess curative power when inoculated into animals artificially infected with a culture of the bacillus pestis. But as the number of experiments carried out was insuflicient to form a definite opinion, and as I had no opportunities to carry out further experiments in Hyderabad, I dropped the subject and did not publish the results. Last September, through the kindness of Professor Dunbar, I was enabled to repeat these experiments at the Hygienic Institute, Hamburg, and the results so far obtained seemed to be very encouraging. The method which I have used in preparing the curative powder from the glands of immunised animals is as follows : Healthy rabbits are inoculated with Haffkine's prophylactic fluid to produce slight immunity, just enough to give them a start. For this purpose I use the supernatant fluid after the sediment consisting of bodies of dead bacilli has thoroughly settled down, as I had shown some years ago 2 that the filtrate from Haffkine's fluid is quite as efficacious in producing immunity as are the entire contents thoroughly shaken up together as recommended by Haffkine, and baE further the advantage of not producing local mischief, suet as hard indurated swelling at the seat of injection containing sterile pus. Haffkine's fluid is given subcutaneously on tw( occasions with an interval of about 12 days between th< injections, and when the animals have thoroughly recovered from its effects, say in from 10 to 12 days, they are subsequently inoculated with avery small quantity of a weak culture of living plague bacilli subcutaneously, and again after an interval o 10 or 12 days a larger dose of the weak culture is intro duced. Subsequently a virulent germ is substituted for th weak one, commencing with the small doses and the] gradually increasing the doses, always allowing an interva of about a fortnight between the injections for the animal to recover, till they can withstand the whole quantity 0 plague germs which will grow on a surface of an ordinar Petri dish. I continue these immunisations until at last th animals can withstand one or two loops of very viruler plague germs given intravenously. Lastly, the anima] when quite sound are killed with chloroform, generall 1 Report of a research carried out during the years 1905 and 1906 at the Hygienic Institute, Hamburg. 2 Indian Medical Record, Jan. 3rd and 10th, 1900; Brit. Med. Jour., May 12th, 1900. .5 days after the last intravenous injection, for the >reparation of the powder. Microscopic examination )f the juice of glands of such animals did not reveal .he presence of any bacteria and no growth was obtained on the media inoculated. It is thus seen that the .mmunity in these animals is really produced by the living oacilli and is far higher than the immunity which could be produced by dead bacilli or their products. The production of such an immunity, however, is much more difficult, as one has to be very careful in graduating the doses so as to avoid killing the animals. I find that it generally takes from four to six months to produce such an immunity. After the animals have been killed with chloroform, the glands such as the liver, the spleen, the buboes, and the suprarenals are removed, are finely minced aseptically and spread in thin layers in sterile glass dishes, and are allowed to dry over sulphuric acid in a hot-air chamber at 470 C. for a period of from three to six days. When the material is thoroughly dried it is well ground in a sterile mortar to a fine powder, after which it is placed in small sterile flasks plugged with cotton-wool and replaced in a hot-air chamber for about 24 hours. The powder is now ready for use. It forms a brown cloudy emulsion when well rubbed down in a mortar with warm sterile water and can be easily injected into an animal with a needle of somewhat large calibre. I found that the process of drying the organs with alcohol takes a much longer time in Hamburg than in Hyderabad. I have therefore adopted Klein's 3 method of drying the organs on account of the insufficiency of time at my disposal. I believe that the alcohol method and the use of larger animals would give better results. The results of experiments with the gland powder thus prepared are given in a tabular form. Two strains of the bacillus pestis were used in the experiments, " Blagdon" " and "Bishopsgate." " The virulence of the former is such that one platinum loopful (two milligrammes) is sufficient to kill a white rat (200 grammes) in 24 hours, th of a loop kills a similar animal in three days, yo'otb of a loop in four days, and 1 o 0 oth of a loop in six days ; while the virulence of the latter is such that ilth, yoth, and T1footh of a loop will kill a white rat (200 grammes) in three, five, and seven days respectively. 50 animals on which the curative powder was administered were as follows : 16 guinea-pigs, 24 brown rats, and ten white rats. In my experiments I have selected the introduction of plague virus in a subcutaneous pouch in the case of brown rats and by a cutaneous method in the case of guineapigs and white rats, as the cutaneous method in the case of guinea-pigs and white rats is a certain method of producing the disease, whereas the cutaneous method in brown rats is not so reliable as the subcutaneous pouch. When working with the Blagdon strain the number of recoveries obtained were 50 per cent. in the case of brown and white rats when plague was given in a subcutaneous I pouch to the former and cutaneously to the latter, as two , (Nos. 2 and 3 (white rats) and 6 and 8 (brown rats) ) out of , four animals recovered in both instances. Whereas with , Bishopsgate the recoveries obtained were 66 per cent., as ; eight (Nos. 26, 28, 29, 31, 32, 33, 34, and 36) out of 12 ; brown rats and four (Nos. 37, 38, 39, and 40) out of six r white rats recovered when treated with the remedy. In the r case of guinea-pigs the results were more favourable. Four were infected with Blagdon (recovered 3: Nos. 10, 11, and & i a c u t e ; 12) and 12 with Bishopsgate (recovered 8 : Nos. 13, 15, 16, 17, 19, 20, 21, and 24) cutaneously. Out of these 16 animals 'f 11 recovered, giving a percentage of a little over 68 recoveries. In these animals one loop (two milligrammes) of agar culture of bacillus pestis of about 48 hours' growth eprocured directly from the smears of organs of animals dead n from acute plague was used in every case and the curative , powder administered varied from 100 to 250 milligramme s doses. In nearly half of the cases the curative powder was f given 24 hours after the infection and in others half an hour , after the infection. When plague was given subcutaneously two brown rats the recoveries obtained were 50 per cent., as efour (Nos. 43, 44, 46, and 47) out of eight animals inoculated is with Blagdon recovered. Out of eight brown rats six had s kth of a loop (Blagdon) subcutaneously and 150 milligrammes .y of the powder about half an hour after the infection and the remaining two brown rats had i sth of a loop (Blagdon) and 3 Klein's Preliminary Report to the Local Government Board, December, 1905.
doi:10.1016/s0140-6736(01)10857-3 fatcat:bobjhh3jbnb5tad3cz5udyfj7i