658 As it is most important in the case of the study of anaerobes to be certain that one is not dealing with mixed cultures, I may briefly relate the steps in the isolation of the bacillus. For convenience I have given it the name B. tumefaciens. From the salicin broth a blood agar plate was on Oct. 6th, 1918, inoculated on the surface. On the 10th a growth, apparently pure, consisting of colonies which were a little flatter than those of typical B. rvelchii was present--the.symbiotic cocco-bacillus did not grow under the conditions of anaerobiosis. Films showed thickish Gram- positive bacilli and many thread and pleomorphic forms. From the blood agar plate a meat broth tube was inoculated, and in it the bacilli flourished and developed spores. A portion of the fluid was pipetted off. boiled, and added to lactose brain. In the lactose brain the spores developed, producing much acid and gas. After 48 hours numerous thick Gram-posi- tive bacilli with central and subterminal spores were found. When transfers were made from the lactose brain into tubes containing sugars it was found that the bacillus fermented glucose, loevulose, galactose, maltose, lactose, saccharose, inulin, starch, glycerine, salicin, with the production of acid and gas. No fermentation of mannite or dulcite was mani- fested and there was no growth on an agar slope under aerobic conditions. From the lactose brain the culture was passed as follows -> lactose brain -> blood agar plate -> lactose brain broth -> 0’2 percent, glucose agar shake plate culture. In glucose agar plate the surface growth con- sisted of a fine film, the deep colonies were charac- teristic and were of varied form ; their shape was often reniform, often that of a whetstone, and often like the ace of hearts-the " eceurs jaunes " of Weinberg. Many of the colonies had a well-defined even edge, whilst others con- sisted of a meshwork of arborescent threads. The most typical colony was one with the edge well defined for part of the way round, but with a bouquet of filaments radiating from the centre and bursting through the periphery at other parts. The grenade form was very common. In short, the aspect of the colonies was such as are so well figured for Vibrion septique by Weinberg and Seguin 2 in Planche II. of their book. I have satisfied myself that the variation of the colony is not due to a mixture of micro-organisms. I have been able, by means of a Pasteur pipette, to take out a large colony with a well-defined and complete periphery and from it grow colonies which were mainly arborescent in appearance. The colonies in glucose agar were found to consist of bacilli with oval spores. A colony containing such bacilli was added to broth and boiled and then planted into meat broth. The fermentative characters of the bacilli in the meat broth corresponded to those already described, and it was this meat broth tube that was the source of the bacilli used in our experiments on animals. Animal experiments.—A full-grown guinea-pig was inocu- lated with 1 c.cm. of a 48 hours’ meat broth culture of the B. tumefaciens. The injection was made subcutaneously and into the muscles of the right thigh. The animal died within 24 hours. At the post-mortem examination a large tumour was found in the right groin. The swelling con- sisted of a pink jelly-like mass of solid oedema ; the muscles underneath contained gas and showed haemorrhages. The abdominal wall on the affected side was reddened both inside and outside. From the oedematous tissue, muscle and liver, bacilli having the characters of B. tumefaciens were recovered. The bacilli on the under surface of the liver contained some thread forms which were in length at least seven times the diameter of a red blood cell. I should mention that in the tissues as well as in culture B. tuniefaciens is actively motile, and that it readily forms" granulose " granules, especially in lactose brain broth. Moreover, it is a little slenderer than B. melekii, and the ends are rounded. In glycerine broth and in some other media the bacillus is more or less spindle- shaped with pronounced polar staining. In glycerine and inulin and in lactose brain clostridial spores predominate. On account of the rapidity with which the bacillus pro- duces a huge oedematous tumour or swelling it seemed appropriate to give it the name B. tumefaciens. Similar experiments showed that the bacillus was patho- genic to rabbits, and that Weinberg’s three sera afforded some degree of protection against the immediate action of .B. ttcnaefaciens. One of the rabbits became paralysed in the hind limbs. Position of the B. tumefaciens in the Classification of Anaerobes. At this point I may briefly summarise the outstanding characteristics of this bacillus, so that they may be compared with those of other anaerobic bacteria of war wounds. The B. tumefaeiens is an actively motile Gram-positive bacillus. In morphological characters it resembles the Vibrion septique, but it is easier to cultivate and forms spores, especially in carbohydrate media, with the utmost facility. Like the Vibrion septique, it often shows swollen citron forms, long threads, and" granulose " bodies. It is essentially a saccharolytic bacillus and fails to digest either coagulated egg albumen or serum. In litmus milk it produces acid and gas. The clotted casein is not digested and remains red in colour-i.e., there is no bleaching. Spores were found in the milk culture. The deep colonies in 0’1 per cent. glucose agar resemble those of the Vibrion septique. In fermenting saccbarose, inulin, and glycerine it resembles some strains of B. welchii and differs from Vibrion septique, but resembles the latter in fermenting salicin. It is far more active in its action on sugars than the two strains of B. cedenaatiens with which I have compared it. In meat broth culture the fragments of muscle become pink in colour and no digestion occurs. It hasmolyses red blood cells. I have compared the characters of B. tumefaciens with those of the various bacteria described in war wounds by French, German, English, and American bacteriologists, and I have come to the conclusion that I am dealing with a new species. There are resemblances between the B. tumefaciens and Aschoff’s " I Gascadembazillus, but there are at least the following points of difference : (1) B. tumefaoiens is actively, the" Gasoedembazillus " feebly, motile ; (2) spores are more readily formed by the B. tumefaciens both in the animal body and in culture ; (3) the lesions caused by the ’’ Gascedem- bazillus " are characterised by a clear gelatinous oedema. Those produced by the B. tumefaciens resemble those of the Vibrion septique in being hæmorrhagic. Further study, especially of the toxin formed by the bacillus and of the agglutinins produced by it and of its response to the agglutinins specific for other bacteria, will be required to determine its position among the pathogenic anaerobes ; in the meantime I think I am justified in giving it the name Bacillus tumefaciens. References.—1. Vide Wilson and Steer: Brit. Med. Jour., Nov. 23rd, 1918, p. 568. 2. La Gangrene gazeuse, by M. Weinberg et P. Seguin, Masson et Cie, Paris, 1918. Medical Societies. ROYAL SOCIETY OF MEDICINE. SECTION OF ANÆSTHETICS. Exhibitiort of Cases. A CLINICAL evening was held on April 4th, Dr. LL. POWELL, the President, in the chair. Mr. ASHLEY DALY reported a case in which direct massage of the hpart restored the circulation and respiration when both had ceased in the course of an abdominal operation upon a patient in a state of severe shock.-The PRESIDENT quoted a case of successful heart massage during laparotomy upon a middle-aged man with rigid abdomen. Dr. F. E. SHIPWAY related a case in which in a feeble man of 60 collapse and heart failure ensued upon drawing the stomach out of the abdomen. Heart massage was successful in restoring the circulation. Mr. R. E. APPERLY read notes of a case in which heart failure followed change of position. The patient was a stout woman anaesthetised with ’’ gas and ether," followed by open ether, for laparotomy. When the Trendelenburg position was adopted pulse and breathing ceased. These were restored by artificial respiration and the horizontal posture. The operation being nearly completed the Trendelenburg position was tried again, when the same sequel of events resulted. The PRESIDENT related a case in which twilight sleep " and spinal analgesia were successfully employed for Cæsarean section on a woman sufeering from influenza and double pneumonia.
Transcript
658
As it is most important in the case of the study ofanaerobes to be certain that one is not dealing with mixedcultures, I may briefly relate the steps in the isolation of thebacillus. For convenience I have given it the nameB. tumefaciens. From the salicin broth a blood agar platewas on Oct. 6th, 1918, inoculated on the surface. On the10th a growth, apparently pure, consisting of colonies whichwere a little flatter than those of typical B. rvelchii waspresent--the.symbiotic cocco-bacillus did not grow under theconditions of anaerobiosis. Films showed thickish Gram-positive bacilli and many thread and pleomorphic forms.From the blood agar plate a meat broth tube was inoculated,and in it the bacilli flourished and developed spores. A portionof the fluid was pipetted off. boiled, and added to lactosebrain. In the lactose brain the spores developed, producingmuch acid and gas. After 48 hours numerous thick Gram-posi-tive bacilli with central and subterminal spores were found.When transfers were made from the lactose brain into tubescontaining sugars it was found that the bacillus fermentedglucose, loevulose, galactose, maltose, lactose, saccharose,inulin, starch, glycerine, salicin, with the production of acidand gas. No fermentation of mannite or dulcite was mani-fested and there was no growth on an agar slope underaerobic conditions. From the lactose brain the culture was
passed as follows -> lactose brain -> blood agar plate ->lactose brain broth -> 0’2 percent, glucose agar shake plateculture. In glucose agar plate the surface growth con-
sisted of a fine film, the deep colonies were charac-teristic and were of varied form ; their shape was oftenreniform, often that of a whetstone, and often like the ace ofhearts-the " eceurs jaunes " of Weinberg. Many of thecolonies had a well-defined even edge, whilst others con-
sisted of a meshwork of arborescent threads. The most
typical colony was one with the edge well defined for partof the way round, but with a bouquet of filaments radiatingfrom the centre and bursting through the periphery at otherparts. The grenade form was very common. In short, theaspect of the colonies was such as are so well figured forVibrion septique by Weinberg and Seguin 2 in Planche II. oftheir book. I have satisfied myself that the variation of thecolony is not due to a mixture of micro-organisms. I havebeen able, by means of a Pasteur pipette, to take out a largecolony with a well-defined and complete periphery andfrom it grow colonies which were mainly arborescent inappearance.The colonies in glucose agar were found to consist of
bacilli with oval spores. A colony containing such bacilliwas added to broth and boiled and then planted into meatbroth. The fermentative characters of the bacilli in themeat broth corresponded to those already described, and itwas this meat broth tube that was the source of the bacilliused in our experiments on animals.Animal experiments.—A full-grown guinea-pig was inocu-
lated with 1 c.cm. of a 48 hours’ meat broth culture of theB. tumefaciens. The injection was made subcutaneouslyand into the muscles of the right thigh. The animal diedwithin 24 hours. At the post-mortem examination a largetumour was found in the right groin. The swelling con-sisted of a pink jelly-like mass of solid oedema ; the musclesunderneath contained gas and showed haemorrhages. Theabdominal wall on the affected side was reddened both insideand outside.From the oedematous tissue, muscle and liver, bacilli
having the characters of B. tumefaciens were recovered.The bacilli on the under surface of the liver contained somethread forms which were in length at least seven times thediameter of a red blood cell. I should mention that in thetissues as well as in culture B. tuniefaciens is actively motile,and that it readily forms" granulose " granules, especiallyin lactose brain broth. Moreover, it is a little slenderer thanB. melekii, and the ends are rounded. In glycerine brothand in some other media the bacillus is more or less spindle-shaped with pronounced polar staining. In glycerine andinulin and in lactose brain clostridial spores predominate.On account of the rapidity with which the bacillus pro-
duces a huge oedematous tumour or swelling it seemedappropriate to give it the name B. tumefaciens.
Similar experiments showed that the bacillus was patho-genic to rabbits, and that Weinberg’s three sera affordedsome degree of protection against the immediate action of.B. ttcnaefaciens. One of the rabbits became paralysed in thehind limbs.
Position of the B. tumefaciens in the Classification ofAnaerobes.
At this point I may briefly summarise the outstandingcharacteristics of this bacillus, so that they may be comparedwith those of other anaerobic bacteria of war wounds. TheB. tumefaeiens is an actively motile Gram-positive bacillus.In morphological characters it resembles the Vibrion septique,but it is easier to cultivate and forms spores, especially incarbohydrate media, with the utmost facility. Like theVibrion septique, it often shows swollen citron forms, longthreads, and" granulose " bodies.
It is essentially a saccharolytic bacillus and fails to digesteither coagulated egg albumen or serum. In litmus milk it
produces acid and gas. The clotted casein is not digestedand remains red in colour-i.e., there is no bleaching.Spores were found in the milk culture.The deep colonies in 0’1 per cent. glucose agar resemble
those of the Vibrion septique. In fermenting saccbarose,inulin, and glycerine it resembles some strains of B. welchiiand differs from Vibrion septique, but resembles the latter infermenting salicin. It is far more active in its action on
sugars than the two strains of B. cedenaatiens with which Ihave compared it. In meat broth culture the fragments ofmuscle become pink in colour and no digestion occurs. It
hasmolyses red blood cells.I have compared the characters of B. tumefaciens with
those of the various bacteria described in war wounds byFrench, German, English, and American bacteriologists, andI have come to the conclusion that I am dealing with a newspecies. There are resemblances between the B. tumefaciensand Aschoff’s "I Gascadembazillus, but there are at least thefollowing points of difference : (1) B. tumefaoiens is actively,the" Gasoedembazillus " feebly, motile ; (2) spores are morereadily formed by the B. tumefaciens both in the animal bodyand in culture ; (3) the lesions caused by the ’’ Gascedem-bazillus " are characterised by a clear gelatinous oedema.Those produced by the B. tumefaciens resemble those of theVibrion septique in being hæmorrhagic.Further study, especially of the toxin formed by the
bacillus and of the agglutinins produced by it and of itsresponse to the agglutinins specific for other bacteria, will berequired to determine its position among the pathogenicanaerobes ; in the meantime I think I am justified in givingit the name Bacillus tumefaciens.
References.—1. Vide Wilson and Steer: Brit. Med. Jour., Nov. 23rd,1918, p. 568. 2. La Gangrene gazeuse, by M. Weinberg et P. Seguin,Masson et Cie, Paris, 1918.
Medical Societies.ROYAL SOCIETY OF MEDICINE.
SECTION OF ANÆSTHETICS.Exhibitiort of Cases.
A CLINICAL evening was held on April 4th, Dr. LL. POWELL,the President, in the chair.
Mr. ASHLEY DALY reported a case in which direct massageof the hpart restored the circulation and respiration whenboth had ceased in the course of an abdominal operationupon a patient in a state of severe shock.-The PRESIDENTquoted a case of successful heart massage during laparotomyupon a middle-aged man with rigid abdomen.
Dr. F. E. SHIPWAY related a case in which in a feebleman of 60 collapse and heart failure ensued upon drawingthe stomach out of the abdomen. Heart massage wassuccessful in restoring the circulation.
Mr. R. E. APPERLY read notes of a case in which heartfailure followed change of position. The patient was a stoutwoman anaesthetised with ’’ gas and ether," followed byopen ether, for laparotomy. When the Trendelenburgposition was adopted pulse and breathing ceased. These wererestored by artificial respiration and the horizontal posture.The operation being nearly completed the Trendelenburgposition was tried again, when the same sequel of eventsresulted.The PRESIDENT related a case in which twilight sleep "
and spinal analgesia were successfully employed for Cæsareansection on a woman sufeering from influenza and doublepneumonia.
659
Dr. SHIPWAY read notes of a case showing the value ofoil-ether anaesthesia in a case in which a physician hadadvised against operation on the grounds that no aneastheticcould be tolerated. The patient had carcinoma of thestomach and had been the subject of aortic aneurysm forfour years.-He also related a case of Enucleation of Tonsilin a man weighing 24 st., whose physique, huge abdomen,large pendulous cheeks and chin, and very short neck madehim appear a most unfavourable subject for anaesthesia.
Operation was successfully performed under aneasthesiainduced by C.E. mixture and continued by warm chloro-form and oxygen, bromide and morphia and atropine havingbeen used as preliminaries.Mr. H. E. G. BOYLE read notes of a case of Laryngo-
fissure with Removal of Intra-laryngeal Growth performedunder gas and oxygen. Morphia and atropine were
used beforehand and gas and oxygen admitted througha catheter passed into the tracheotomy tube, tracheotomyhaving been performed under gas and oxygen, withre-breathing.
____ ___
Reviews and Notices of BooksSanitation P1’aoUoally Applied. By H. B. WOOD, M.D.,
D P. H. With numerous illustrations. London: Chapmanand Hall ; New York : John Wiley and Sons. Pp. 474.13s. 6d.
THE object of this book is to supply information on thepractical application of the principles of public health andsanitation. It is intended primarily for the use of healthofficers and students of public health, but the authorexpresses the hope that employers, teachers, and municipalofficials may also gain practical directions in the art of
healthy living from a perusal of its pages. Some indica-tion of the scope of the.work may be given by enumera-tion of the subjects treated in the 12 chapters which itcontains. The headings of the chapters are as follows : theneed for public health work, statistics, the control ofcommunicable diseases, child welfare, school hygiene, purefoods, clean milk, water-supplies, sewage disposal, hygieneof the home and factory, the destruction of insects whichtransmit disease, and the methods which may be adopted ineducating the public in sanitary matters.The book is obviously intended for American rather than
British readers, and contains relatively few references to thework of sanitarians in this country. The chapter on schoolhygiene contains some helpful observations on the medicalinspection of school children, and that on " clean milk is san excellent, if somewhat short, exposition on a subjectwhich is at last receiving much needed attention in
England. In reference to the standardising of milk whichis practised in certain parts of the United States of America,the author gives the standards recommended by the NewYork Milk Committee, which briefly are as follows :-Class A. Certified milk or its equivalent.-The term certified" milk
should be limited to milk produced in conformity with the require-ments of the American Association of Certified Milk Commissions,while the term "equivalent" indicates that produced at dairies sub-jected to periodic inspection and the products to frequent analyses.The cows must be properly fed and watered, be free from tuberculosis,as shown by the tuberculin test and veterinary examinations, and fromother diseases or conditions liable to deteriorate the quality of themilk. They must be kept in a state of cleanliness in clean, well-ventilated cow-houses. All persons coming in contact with the milkmust be healthy and free from typhoid fever, tuberculosis, diphtheria,scarlet fever, septic sore throat, and other diseases which mav infectthe milk. Pure water must be provided at the farm. The milk must’be drawn under all sanitary precautions to prevent contamination, beimmediately strained andjpooled, placed in sterilised bottles, and main-tained at a temperature at or below 500 F. It shall not contain morethan 10,000 bacteria per c.cm.
Class 8. Inspected milk.-This class includes clean raw milk fromhealthy tuberculin-tested cows and from dairies that score not less than70 on the Government score card. The cows are fed, watered, andmilked under good conditions The dairymen must exercise cleanli-ness and be free from diseases which may be milk-borne. The milk isdelivered in sterilised containers and kept at a temperature not greaterthan 500 until it reaches the consumer. It should contain less than100,000 bacteria per c.cm. Pasteurisation is optional.
Class C. Pasteurised Nt.—Milk from dairies not able to comply withthe A and B requirements should be pasteurised, under official super-vision, and then sold as "pasteurised milk." Milk for pasteurisingshould be kept at not over 500 while in transit to the plant. Afterpasteurisation it is cooled at once to a temperature of 50° or belowand maintained there. No cows in any way unfit for the productionof milk for use by man shall be permitted to remain in any Class C
dairy. The milk before pasteurisation should contain not over
1,000,000 bacteria per c.cm., and not over 50,000 after pasteurisation.It should not show colon bacilli in 1 c.cm. It should be delivered tothe consumer within 48 hours after pasteurisation. Repasteurisationis prohibited.
Class D. Milk not suitable for drinking pecrposes.-Mitk containingover 1,000,000 bacteria per c.cm., or from farms scoring less than40 per cent., may be allowed for cooking and industrial purposes onlyafter being pasteurised or heated to a higher degree. It should bedelivered in distinctive containers and must not be sold for drinking.The American system of grading milk possesses many
advantages. It makes farmers and dairymen keenlyinterested in the productiun of clean wholesome milk, andensures higher prices to those who are successful in this
respect. No doubt it would be preferable if all milk sold tothe public reached a Class A standard of excellence, butthis is not practicable under existing conditions of produc-tion, and it is very important that the public should knowwhat they are buying, and be assured that, if they canafford the price of the higher grades of milk, they receivewhat they are paying for.
Dr. Wood’s book contains much valuable information in a
relatively compact form, and should be useful to all thoseinterested in the practical side of hygiene and public health.
Aids to 81.l1’gery. By JOSEPH CUNNING, M.B., B.S. Melb.,F.R.C.S. Eng., Surgeon to the Royal Free Hospital ; andCECIL A. JOLL, M.S. Lond., F.R.C.S. Eng. Fourthedition. London: Bailliere, Tindall, and Cox. 1919.Pp. 420. 4s. 6d. net.
THIS is the fourth edition of a well-known volume of thefamiliar Aids" " series, carefully revised and broughtthoroughly up to date. The principal new feature is theaddition of a description of war injuries and infections,together with their treatment under the appropriateregional headings. Compactness, a simple arrangement,and singularly clear headings combine with a terse style tomake this book a valuable help in revision. As an initial
text-book its value is doubtful; it lays no claim to fulfil thisfunction. But as a rapid aid to memory it will continueto fill the place it has held in many a student’s pocket.
Elements of Surgical IJiagnosis. By Sir ALFRED PEARCEGOULD, K.C.V.O., and ERIC PEARCE GOULD, F.R.C.S.Fifth edition. London : Cassell and Co., Ltd. 1919.Pp. 722. 12s. 6d.THIS book has now passed into its fifth edition with many
reprints. Owing to the omission of some of the older methodsof diagnosis, now fallen into disuse with the advent of X raysand other more exact means, the publication is slightlyreduced in size. This is an unusual and, to students, awelcome change. There are 16 excellent X ray prints-those in the genito-urinary section being of special merit.The authors have set out to perform a great task-to initiatethe student in accurate methods of examination and clinical
diagnosis, and to stimulate him by demonstrating the essentialvalue of these methods in dealing with every surgical condi-tion. With a wealth of detail, a free use of headings anditalics, and an insistence on the necessity for meticulous carein dealing with the differentiation of each type of disease orinjury, they go far to attain their object.The book will continue to be of inestimable value to the
student at the outset of his clinical career-to use wordsemployed just above. Many who have passed that stagewould do well to renew their clinical youth in its pages.
An Int1’oduotil’n to the Study o Biologioal 4chemistry. ByS. B. SCHRYVER, D.Sc. Lond. London and Edinburgh :T. 0. and E. C. Jack. Pp. 340. 6s.THE chemical processes involved in metabolism form the
principal theme of this book, and only one branch, but, ofcourse, an important branch, of the study of biologicalchemistry is presented. The student of biochemistry mustbegin with a knowledge of the structure of the materialswhich compose the organism and the nutritional elementswhich maintain and build up the organism, and Dr. Schryverhas been successful in giving a clear exposition of thissection of his subject. The monographs on the hydrolysisof proteins and separation of the amir. -cids, and on theconstitution of the proteins, their c- incation, prepara-tion, and general properties, form good examples of hisintention. The book furnishes a very useful opening to thestudy of biochemistry.
