PROCEEDINGS OF LOCAL BRANCHES OF THESOCIETY OF AMERICAN BACTERIOLOGISTS

TEXAS BRANCHSECOND MEETING, THE UNIVERSITY OF TEXAS MEDICcA BRANCH,

GALVESTON, TExAS, MAY 2, 1942

SOME ASPECTS OF NUTRITIONAL VARIATIONOF THE GONOCOCCUS. C. E. Lankford,Dept. of Obstetrics and Gynecology, TheUniversity of Texas Medical Branch, Gal-veston, Texas.A type of gonococcus is described which

does not grow on autoclaved culture media,including Proteose no. 3-hemoglobin agar(Bacto) and glucose-starch agar, except inthe presence of certain other bacteria orwith the addition of extracts of liver, yeast,or blood or similar substances. Thesestrains occur with considerable frequency,so that failure to detect them constitutes aserious defect in cultural diagnosis.The addition of one part of liver extract,

sterilized by filtration or preserved undertoluene, to 200 parts of agar permits luxuri-ant growth of these deficient strains. Suchstrains undergo spontaneous mutation toproduce variants capable of synthesis of thedeficient nutrilite and resembling the"normal" strains of gonococcus in all otherrespects. The reverse of this variation hasbeen observed in vivo.The factor involved is relatively simple,

dialyzable, alcohol-soluble, and is destroyedby autoclaving. It ia somewhat acid-labileand is not precipitated by mercuric acetateor adsorbed by activated charcoal at pH 3.Attempts to replace it by a number of knownpure nutrilites, including cozymase and co-carboxylase, have been unsuccessful.

All strains of the gonococcus tested growwell on casein-hydrolysate-starch agar onthe addition of 1/200 liver extract. Onlythe "normal" strains grow if the extract isautoclaved.

NICOTINAMIDE-CONTAINING NUTRILITESFOR HEMOPHILUS INFLUENZAE. WendellGingrich, Dept. of Bacteriology, andFritz Schlenk, Dept. of Preventive Medi-cine and Public Health, The University

of Texas Medical Branch, Galveston,Texas.Codehydrogenases I and II, which are

essential as coenzymes of hydrogen trans-port in carbohydrate metabolism, satisfythe V-factor requirement of Hemophilusinfluenzae and H. parainfluenzae (Lwoff andcLwoff, 1936). We have observed growth ofHemophilus influenzae in a simple peptonemedium containing 2 X 1I- gamma mol ofCodehydrogenase I (cozymase) per ml. ofmedium. Several derivatives and splitproducts of cozymase were also tested foractivity in promoting growth of H. influ-enzae. Codehydrogenase II, dihydrocozy-mase, acid-treated dihydrocozymase, desa-minocozymase and nicotinamide nucleosidesupported growth in concentrations of thesame order of magnitude as cozymase. Thestructural units of the cozymase molecule;i.e., nicotinamide, d-ribose and adenylicacid, are inactive. Therefore, the nico-tinamide-pentose linkage of the nucleosideis the minimum for satisfying the V-factorrequirement of H. influenzae.

AcCESsORY GROWTH FACTOR REQUIREMENTSOF BRUCELLA. N. B. McCullough andLeo A. Dick, Brucellosis Research Labora-tory, Clayton Foundation, The Uni-versity of Texas, Austin, Texas.

NEWER KNOWLEDGE OF VIRUS ENCEPHALI-TIDES IN TExAS. J. V. Irons, S. W. Bohls,and Dorothy Albert, Bureau of Labora-tories, Texas State Department of Health,Austin, Texas.

DISTRIBUTION OF VECTORS AND SPIRO-CHETES OF RELAPSING FEVER IN TEXAS.J. L. Terrell, T. H. McGregor, and A. K.Mayes, Bureau of Laboratories, TexasState Department of Health, Austin,Texas.

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INVESTIGATIONS ON TYPHUS FEVER INTEXAS. Ludwik Anigstein and MaderoN. Bader, Dept. of Preventive Medicineand Public Health, The University ofTexas Medical Branch, Galveston, Texas.The work, still in progress, has been

directed along the following lines:1. Serological survey of domestic animals

as potential links in the epidemiologyof local typhus.

2. Mass examination of various ectopara-sites of dogs, cattle and hogs as possiblevectors of typhus.

3. Examination of wild rats from epidemicfoci and endemo-sporadic areas oftyphus.

4. Isolation of typhus strains from humansources.

5. Comparative study of the isolatedstrains of different origin, includinglouse-borne typhus.

Several hundred sera of dogs, cattle andhogs were examined for Weil-Felix reactionwith positive results (1:160 up 1:320) in aproportion of cases. Striking differenceswere found in dogs naturally infested withticks (Rhipicephalus sanguineus) (94.7%),as compared with tick-free dogs (30%).Similar differences were found in cows in-fested with ticks (Amblyomma maculatumand A. americanum) and in tick-free cattle.Hog-sera gave in 18% of the cases a strongpositive Weil-Felix reaction.

Inoculation of batches of ticks and hog-lice into guinea pigs resulted in fever andoccasionally in scrotal reactions. Some im-munity against murine typhus was noted inguinea pigs inoculated with hog-lice.Two strains of murine typhus were iso-

lated directly from wild rats from an epi-demic focus of typhus and also from anendemo-sporadica typhus area. Other ty-phus strains were recovered from humanpatients. A comparative study of all theisolated strains including louse-borne ty-phus secured recently from Spain is underway.On the basis of the above results the

authors are inclined to believe that domesticanimals and their ectoparasites play anadditional r6le in the epidemiology ofrickettsial diseases in Texas.

ORGANISMS FOUND IN CASES OF AcuTOPHTHALMIA. Charle8 A. Parker, De-partment of Ophthalmology, The Uni-versity of Texas Medical Branch, Gal-veston, Texas.Of the 158 cases submitted to the labora-

tory for bacteriological examination, 42cases presented the clinical picture of acuteconjunctivitis. Following is a list of theorganisms found in these 42 cases:

cases

Neisseria intTracellulari8........ 18Staphylococcus ................. 18Diplococcus pneumoniae......... 4Koch-Weeks bacillus........... 2

We have seen two cases of trachoma.The location of the inclusion body in thetrachomatous cell is usually'unipolar; how-ever, one of these cases presented an un-usual picture in that bi-polar inclusionbodies were present with one seeming toinvade the nucleus of the epithelial cell.

DIFFERENTIAL DIAGNOSIS OF YELLOWFEVER, EPIDEMIC JAUNDICE, AND WEIL'SDISEASE. A. Packchanian, Departmentof Preventive Medicine and PublicHealth, The University of Texas MedicalBranch, Galveston, Texas.

A CASE REPORT OF COCCIDIOIDAL GRANU-LOMA. Leo J. Peters, Dept. of InternalMedicine, The University of Texas Medi-cal Branch, Galveston, Texas.

THE INCIDENCE OF PARASITIC FUNGI INGALVESTON. Mildred B. John, Dept. ofBacteriology, The University of TexasMedical Branch, Galveston, Texas.

NOTES ON FUNGUS SEROLOGY. William B.Sharp, Dept. of Bacteriology, The Uni-versity of Texas Medical Branch, Gal-veston, Texas.Immune response is observed to an anti-

gen from molds prepared as previouslydescribed by extraction into saline andsalting out. Circulating antibody reacheshigh titer. Evidence on intracellular anti-body is as yet inconclusive. Lethal anddermal toxicity for guinea pigs is evident,with suggestion of increased sensitiveness tosuch action on immunization.

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FACTORS AFFECTING THE ACTIVITY OFPSEUDOMONAS-PHYTOMONAS FLUORES-CENT BACTERIA IN SOIL. Roland B.Mitchell and Francis E. Clark, Bureau ofPlant Industry, United States Depart-ment of Agriculture, Greenville, Texas.Investigations have been made to deter-

mine the extent to which activity of com-ponent groups within the soil microfloracan be controlled. This report considersthe activity of blue-green fluorescent bac-teria, as revealed by the populations deter-mined with an asparagin-medium, dilutiontube method (Clark, 1940), under differingsoil conditions.

In unamended field soil, this group con-stituted a minor fraction of the total micro-flora; the group increased slightly in num-bers and in relative importance followingtillage and precipitation. Activity of thisgroup was increased moderately in soilunder growing cover crops, and markedly,following applications of organic materials.Plant materials added to soil increasednumbers of blue-green fluorescent bacteriaover longer periods than did animal ma-nures. In soil receiving either 3.0 per centchopped green plant material, or washedfungus mycelium from laboratory cultures,fluorescent types were increased to severalhundred millions per gram. In suchamended soils, this bacterial group consti-tuted as large a percentage of the totalmicropopulation as it commonly does onthe root surfaces of plants. Fluctuations innumbers of blue-green fluorescent bacteriaon plant roots, both with different ages andspecies of plants, and with different methodsof stem and root mutilation, are discussed.

ANTIBIOSIS IN THE ELIMINATION OF PHYMA-TOTRICHUM OMNIVORUM SCLEROTIA FROMSOIL. Francis E. Clark and Roland B.Mitchell, Bureau of Plant Industry,United States Department of Agriculture,Greenville, Texas.This report considers the elimination of

sclerotia of the cotton root-rot fungus fromsoil. It has previously been demonstratedin this laboratory that such sclerotia areeliminated from soil amended with organicmaterial more rapidly than from unamendedsoil. Current studies show the dependenceof such destruction of sclerotia on microbial

activity, and define certain conditions fa-voring antibiotic effects.Uncontaminated viable sclerotia survived

in sterile, organic-amended soil equally aswell as in sterile unamended soil. In non-sterile, amended soil, incubation tempera-tures favoring general microbial activitywere more destructive to sclerotia; incuba-tions of 20, 120, 280, and 35°C. eliminated12, 30, 72, and 91 per cent, respectively, ofviable sclerotia. At 28°C., soil moisturecontents of 35, 58, and 80 per cent werefound effective in the order named. Thefactors of aeration and soil reaction werealso considered. The effects of differentorganic amendments were compared underconstant conditions of incubation. Ma-terials with narrow C/N ratios providedequally as great destruction of sclerotia asdid materials with wide ratios; it was un-necessary to use soil amendments whichwould not meet good crop nutrient require-ments. Types of microbial activity en-countered and the application of antibiosisin field sanitation are briefly discussed.

A NEW STAINING PROCEDURE FOR THEBRUCELLA OPSONOCYTOPHAGIC TEST. LeoA. Dick and N. B. McCullough, BrucellosisResearch Laboratory, Clayton Founda-tion, The University of Texas, Austin,Texas.

THE GLANDERS ORGANISM WITH REFERENCETO CELL INCLUSIONS. Gordon Worley andGerald Young, Dept. of Preventive Medi-cine and Public Health, The Universityof Texas Medical Branch, Galveston,Texas.The irregular staining of the glanders

organism is mentioned frequently in theliterature with different interpretationsconcerning this property. Inasmuch as weencountered no explanation which seemedadequate, we have made a morphologicalstudy of five strains of the Malleomycesmallei.These organisms were found to have un-

stained areas when such dyes as gentianviolet, methylene blue, or acid fuchsin wereemployed. However, when the fat stainssuch as Sudan Black B. or Eisenberg'sfuchsin-iodine were employed, stained gran-ules stood out prominently. We have been

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unable to demonstrate the presence of large part if not entirely to the presencevolutin. within the protoplasm of lipoidal granulesWe may state, then, that the irregular which fail to stain readily with the usual

staining of the glanders organism is due in aniline dyes.

EASTERN PENNSYLVANIA CHAPTER

ONE HUNDRED AND FIFTY-SIXTH MEETING, PHILADELPHIA COUNTY MEDICAL SOCIETYBUILDING, PHILADELPHIA, PA., JANUARY 27, 1942

STUDIES IN SURGICAL BACTERIOLOGY. I.DISTRIBUTION OF BACTERIA AROUND THE

NAILS AND ON THE SKIN IN RELATION TO

DISINFECTION OF THE HANDS. Rtussel H.Fowler, Abington Hospital, Abington, Pa.

STUDIES ON THE NATURE OF THE VIRUS OF

INFLUENZA. Leslie A. Chambers andWerner Henle, Department of Pediatricsand The Johnson Research Foundation,

University of Pennsylvania, Philadel-phia, Pa.

THE MOVING PICTURE "UNSEEN WORLDS."Harry E. Morton, University of Pennsyl-vania, School of Medicine, Philadel-phia, Pa.A 400 foot, 16 mm, black and white film,

with sound, illustrating the principles andsome applications of the RCA electronmicroscope.

ONE HUNDRED AND FIFTY-SEVENTH MEETING, PHILADELPHIA COUNTY MEDICAL SOCIETYBUILDING, PHILADELPHIA, PA., FEBRUARY 24, 1942

A STUDY OF MICROAEROPHILIC ORGANISMSWITH SPECIAL REFERENCE TO THE USE OFA MICROAEROPHILIC INCUBATOR. S.Brandt Rose, Philadelphia General Hos-pital, Philadelphia, Pa.A study of a large number of clinical speci-

mens showed that added carbon-dioxide wasnecessary for the isolation not only of alimited number of organisms like Brucella,but also of many common pathogens, suchas Pneumococcus, Streptococcus pyogenes,Hemophilus influenzae, etc. Experienceshowed that it was impossible to predictwhich specimen would require such an en-vironment. Hence it is recommended thata non-ventilated carbon-dioxide incubator*be employed routinely in diagnostic lab-oratories. Evidence was presented to provethat this procedure yielded a significantlyhigher incidence of positive results. It isproposed that the incubation be designatedas capneic (from the Greek, kapnos, meaningcarbon-dioxide, as in acapnia) and that anorganism which needs such incubation becalled a capnophile.

* The author is indebted to Mr. W. H.Reynolds of the American Instrument Co.,Silver Springs, Md., for providing the in-cubator for experimental purposes.

STUDIES IN THE PREVENTION OF AIR-BORNEINFECTION. Werner Henle, Harriet E.Sommer and Joseph Stokes, Jr., Children'sHospital, 18th and Bainbridge Sts., Phila-delphia, Pa.Animal experiments were performed in a

hospital ward in order to test and to com-pare the effect of ultraviolet irradiation andof propylene glycol vapor on air-bornespread of respiratory diseases by dropletnuclei under controllable conditions. Whenheavy concentrations of air-borne hemolyticstreptococci of Lancefield's group C wereatomized, most of the mice died fromstreptococcal pneumonia and septicemia inall cubicles regardless of the distance fromthe bacterial atomizer. Under the influenceof ultraviolet irradiation all mice but a fewin the atomizer cubicle survived. Propy-lene glycol vapor prevented infection com-pletely. If a low concentration of group Cstreptococci was used all mice including thecontrols survived and no organisms could berecovered from the lungs on the 8th day.However, in other experiments it could bedemonstrated that a streptococcal carrierstate had been induced in the control miceas shown by a provocative infection with thevirus of influenza A 8 to 10 days after the ex-posure to the contaminated air. All mice

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died from influenza but only the controlgroups now revealed also the streptococcus.Similar results were obtained when the virusof influenza A was atomized. While all thecontrol animals died, propylene glycol vaporprevented death in all cubicles and underultraviolet irradiation only a few mice diedin the atomizer cubicle.

MICROBIAL ANTAGONISM AND BRUCELLAABORTUS. Walter Kocholaty, School ofVeterinary Medicine, University of Penn-sylvania, Philadelphia, Pa.Of a number of antibacterial preparations

tested, three substances appeared to behighly bacteriostatic and bactericidalagainst Brucella abortus, in vitro. Thosesubstances are: pyocyanase, streptothricin(an antibacterial substance produced by an

Actinomyces of the A. lavendulae type, iso-lated by the writer from soil and dust), and"penatin" (a newly discovered antibac-terial substance produced by Penicilliumnotatum). While penicillin, also a product

from Penicillium notatum has only very

weak antibacterial properties against Bru-cella abortus, and practically none againstEscherichia coli, penatin is highly bacterio-static and bactericidal against those twoorganisms and even more powerful againstgram-positive micro-organisms, such as

Staphylococcus aureus. Evidence has beenoffered that penicillin and penatin are by no

means identical. As the crude culture fil-trate of the mold producing penatin con-

siderably surpasses in its antibacterial ac-

tion streptothricin and pyocyanase, most ofthe experiments conducted were carried outwith the above-mentioned mold. The bestculture conditions for the production ofpenatin were established. Using optimalculture conditions, 0.02-0.01 ml of the crudeculture filtrate in 10 ml of agar will eitherstrongly inhibit or completely suppress thegrowth of B. abortus and an even smalleramount is sufficient to impede the growth ofS. aureus.

THE ONE HUNDRED AND FIFTY-EIGHTH MEETING, PHILADELPHIA COUNTY MEDICAL SOCIETYBUILDING, PHILADELPHIA, PA., MARCH 24, 1942

SOME PRACTICAL APPLICATIONS OF BAC-TERIOLOGY TO MEDICAL NURSING. CarlJ. Bucher, Jefferson Hospital, Philadel-phia, Pa.

MOLD INHIBITION IN VARIOuS FOOD PROD-UCTS THROUGH THE USE OF INHIBITORY

CHEMICALS. D. K. O'Leary, E. I. du Pontde Nemours & Co., Experimental Station,Wilmington, Del.

THE GERMICIDAL PROPERTIES OF SOAPS.Werner Leszynski, Research Department,Fels Soap Co., Philadelphia, Pa.

ONE HUNDRED AND .FIFTY-NINTH MEETING, PHILADELPHIA COUNTY MEDICAL SOCIETYBUILDING, PHILADELPHIA, PA., APRIL 28, 1942

A SIMPLE TuIBE BATTERY FOR USE IN BAC-TERIAL AIR ANALYSIS. W. F. Wells,Laboratories for the Study of Air-borneInfection, University of Pennsylvania,School of Medicine, Philadelphia, Pa.The quantitative accuracy of enrichment

methods of isolating Escherichia coli fromwater, and Streptococcus viridans frombreathed air, is statistically limited by thenumber of portions tested. For thesemethods to be useful in sanitary analysis itis necessary to simplify handling of a largenumber of tubes. To obviate the necessityof using cotton plugs and unwieldy racks in

water analysis, Macrady substituted a

single cover to a battery of fermentationtubes. Wassermann tubes have been foundconvenient in air analysis for distributingdilutions of collecting liquid. Fifteen ofthese tubes can be circled around a no. 8rubber stopper. Two rubber stoppers heldapart by a short length of glass tube form a

convenient spool around which to assemblefifteen tubes held against the stoppers byrubber bands. A half-pint cylindricalaluminum measuring cup fits snugly over thebattery but leaves enough of the tubes show-ing to observe color changes in the medium.

Inoculation of the tubes and transference

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to selective gentian violet blood agar andrecording results are greatly simplified.

THE BOERNER-LUKENS WASSERMANN TESTWITH SPINAL FLUID WITH SPECIAL REFER-ENCE TO THE USE OF EGG ALBUMIN. FredBoerner, Marguerite Lukens, and AliceEllis, Graduate Hospital, Philadel-phia, Pa.

THE BOERNER-JONES-LUKENS MACROFLOC-CULATION TEST WITH SPINAL FLUID.Fred Boerner, Marguerite Lukens, andAlice Ellis, Graduate Hospital, Phila-delphia, Pa.

SPIROCHETAL ANTIGENS IN THE SERUMDIAGNOSIS OF SYPHILIS. John A. Kolmer,Temple University School of Medicine,Philadelphia, Pa.

Complement fixation in syphilis withspirochetal antigens is a group reaction as

positive reactions occur with antigens pre-

pared not only of cultures of the Reiter andother strains of Treponema pallidum butT. microdentium and T. macrodentiumas well.Antigens prepared from cultures of these

spirochetes give a small and varying per-centage of nonspecific or falsely-positivecomplement fixation reactions with the sera

of normal individuals and those with tuber-culosis, malignant diseases and febrile in-tercurrent illnesses ascribed to the presenceof natural spirochetal antibody.

Spirochetal antigens give a particularlyhigh percentage of non-specific or falsely-positive complement-fixation reactions inleprosy and malaria just as tissue or lipoidalantigens yield a high percentage of positiveWassermann and flocculation reactions inthese diseases.The spinal fluids of nonsyphilitic indi-

viduals, however, do not give nonspecific or

falsely-positive complement-fixation reac-

tions with spirochetal antigens which is as-

cribed to the absence of natural spirochetalantibody in them.

Spirochetal antigens prepared from viru-

lent T. pallidum recovered from acute testi-cular syphilomas of rabbits give a higherpercentage of positive complement fixationreactions with syphilitic sera and a lowerpercentage of falsely-positive reactions withnormal sera than antigens prepared fromcultures of alleged T. pallidum.Available evidence, based upon the results

of absorption tests, indicates that thesyphilis reagin giving positive Wassermannand flocculation reactions is entirely sep-

arate and distinct from the natural spiro-chetal antibody in normal sera or the ac-

quired spirochetal antibody in syphilisyielding positive complement-fixation re-

actions with spirochetal antigens.The value of spirochetal antigens pre-

pared from the Reiter strain in the serum

diagnosis of syphilis is as yet undecided. Inthe Washington serological survey the Kol-mer spirochetal antigen was less sensitivethan the Kolmer tissue or lipoidal extract(C. L.). The Eagle Wassermann reactionusing tissue antigen, however, was lesssensitive than a spirochetal antigen.A mixture of spirochetal and tissue or

lipoidal antigens has been found more

sensitive in complement-fixation tests withsyphilitic sera than spirochetal antigenalone but less sensitive than tissue or lip-oidal antigen alone. The mixture has alsogiven a smaller percentage of nonspecific or

falsely-positive reactions with normal sera

than the spirochetal antigen alone.

THE MOVING PICTURE "A LECTURE ON THE

SPIROCHETES." Harry E. Morton, Uni-versity of Pennsylvania, School of Medi-cine, Philadelphia, Pa.Three 400 foot reels, 16 mm, black and

white film prepared by Theodor Rosebury,College of Physicians and Surgeons, NewYork. Most of the picture is devoted todark-field photography of the spiralled or-

ganisms of medical importance. The mor-

phological characteristics of all the spir-alled organisms are illustrated and the tech-nic for preparing dark-field preparations isdemonstrated.

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ONE HUNDRED AND SIXTIETH MEETING, PHILADELPHIA COUNTY MEDICAL SOCIBTY BUILD-ING, PHILADELPHIA, PA., MAY 26, 1942

THE PRODUCTION OF ANTIBIOTIC AGENTS BYMICROORGANISMS AND ]TS SIGNIFICANCEIN NATURAL PROCESSES. Selman A.

Waksman, State of New Jersey Agricul-tural Experimental Station, New Bruns-wick, N. J.

VIRGINIA BRANCHUNIVERSITY OF VIRGINIA, CHARLOTTESVILLE, NOVEmBER 1, 1941

TRANSFORMATION OF FIBROMA VIR1US TOTHAT OF INFECTIOUS MYXOMA. RalphB. Houlihan, University of Virginia.

PREPARATION OF SIMPLE VACuuM FLASK.Edward Lavor, Norfolk General Hospital.

THE ACTION OF SULFONAMIDE DRUGS UPONCERTAIN FREE-LIVING PROTOZOA. Fred-erick F. Ferguson, Jane Holmes andEdward Lavor, College of William andMary and the Norfolk General Hospital.

LABORATORY PROCEDURES IN THE DIAG-NOSIS OF CERTAIN FUNGUS INFECTIONS.J. D. Reid and R. A. Holt, Medical Collegeof Virginia.

THEI MULTIPLICATION OF BACTERIA INSTERILE CRABMEAT. Caroling Berry, U.S. Public Health Service Laboratory atCraney Island, Norfolk, Virginia.Cultures of Escherichia coli, Proteus sp.,

Salmonella aertrycke, Salmonella morgani,Eberthella typhi, Shigella dysenteriae (Flex-ner) and Staphylococcus aureus were inocu-lated respectively into autoclaved crabmeatand the rate of growth observed. Signifi-cant increases in the bacterial plate count

were observed with each culture employedwhen the incubation temperature was 25degrees C. or 37 degrees C. At 5 degreesC., however, the bacterial numbers de-creased, although viable organisms were stillpresent in the crabmeat after 15 days at thistemperature.

THE BACTERIAL FLORA OF VIRGINIA MILKAND ITS RELATIONSHIP TO STANDARDS INTHIS STATE. F. S. Orcutt, VirginiaPolytechnic Institute.

INAPPARENT VIRUS INFECTIONS. Norman T.Ashenburg, Craney Island Laboratories.

CURRENT INVESTIGATIONS ON AN IMPROVEDKEY TO THE GENUS BACILLUS. W. B.Coffee, R. J. Fitzgerald and F. S. Orcutt,Virginia Polytechnic Institute.

METHODS FOR THE EXAMINATION OF SHELL-FISH AND SHELLFISH PRODUCING WATERS.Leslie A. Sandholzer, Craney Island Lab-oratories.

THE INFLUENCE OF SEAWATER ON BAC-TERIAL PLATE COUNTS. Marjorie Smith,Craney Island Laboratories.

ROANOKE, VA., MAY 8-10, 1942

STREPTOCOCCI RESISTANT TO SULFONAMIDETHERAPY. George McL. Lawson, Depart-ment of Preventive Medicine and Bac-teriology, University of Virginia, Schoolof Medicine, Charlottesville, Virginia.The favorable report on sulfonamide

therapy of streptococcus infections in gen-eral must be modified in the future by a

more careful identification of groups andtypes to which offending organisms belong.The classification into hemolytic, non-

hemolytic and viridans groups is a time-

honored but scientifically inaccurate divi-sion of the streptococci. Within each suchgroup it is possible to pick out organismswhich are pathogenic but are not affectedby sulfonamide therapy. Among these areStreptococcus fecalis in the so-called viridansgroup and Lancefield's Group B strepto-coccus in the so-called hemolytic group.Case reports are given concerning three

patients infected with streptococci of Lance-field's Group B. Additional reports are pre-sented showing the contrasting therapeutic

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results of sulfonamide therapy in sub-acutebacterial endocarditis due to Streptococcusfecalis and to Streptococcus salivarius.

A SIMPLIFIED METHOD FOR ANAEROBICPLATE CULTURES. William E. Bray andJeanette S. Carter, University of Virginia.

HUMAN INFECTION CAUSED BY SALMONELLASAN DIEGO. William E. Bray, Universityof Virginia.

CULTURAL STUDIES OF STAPHYLOCOCCI Iso-LATED FROM SHELLFISH. Herbert Birtha,Velma Brewington, Alethia Greene, Wil-liam Quivers and Lilly Riddick, HamptonInstitute.

QUANTITATIVE DETERMINATIONS OF MYX-OMA VIRUS IN NASAL WASHINGS, BLOOD,FECES AND URINE AND THEIR RELATIONSTo TRANSMISSIBILITY. Ralph B. Houli-han and George McL. Lawson, Universityof Virginia.

KODACHROME PHOTOMICROGRAPHY. Ken-

neth B. Grim, Department of ClinicalPathology, University of Virginia.There are many methods of obtaining

good photomicrographs in color. The mostcommonly used are those in which the Leicaor Contax cameras are attached to the eye-piece of the microscope with the micro-ibsomiflex or similar photomicrographic attach-ments. Good natural color photographsmay also be obtained by using an adapterring and placing any 35 mm. camera directlyover the eyepiece of the microscope. Wehave found that a 32000 Kelvin light placedabout one foot from the sub-stage mirrorgives the best results. Type A Kodachromefilm for artificial light does not have a verywide latitude and therefore the exposuremust be exact, and the setup once in placemust remain constant for each objective.

Since many photographers already havecameras which take 3j by 4i inch plates, the35 mm. adapter for the kodak Recomarcamera is a valuable adjunct, for it can befitted directly to any of these cameras.

With this adapter and with a bellows typecamera, almost any desired magnificationmay be obtained. It is especially valuablefor obtaining photographs without reduc-tion in the size of the image. Since only thecentral portion of the field is photographed,

a critical focus is easily obtained and there isno blurring of the periphery as sometimesoccurs with other methods. This adapter isquite inexpensive and the photographs ob-tained with it are excellent reproductions innatural color.

NEGRO HEALTH PROBLEMS AND NATIONALDEFENSE. T. W. Turner, Hampton In-stitute.

WAR AsPECTS OF THE POLLUTION PROBLEM.L. A. Sandholzer, U. S. Public HealthService, Norfolk, Va.

STREAM ECOLOGY AND AVAiLABLE FOOD.Leonard I. Katzin, U. S. Public HealthService, Norfolk, Va.A study of the principal streams of the

Ohio River has shown certain differences be-tween streams and between portions of thesame streams, in protozoan and algal plank-ton populations, and in fish populations.It can be shown, other conditions beingequal, that these differences are closely re-

lated to the amounts of food available to theplanktons, as a function of the amount of or-

ganic material introduced into the stream,either by natural means or through thedirect agency of man. In general, on thisbasis, great differences between the streamsof this basin are a reflection of differences inthe amount of organic "fertilizer" available.

It has been possible to differentiate threemain types of plankton: (a) a highly r¢-sistant, "thrifty" group, able to withstandlow temperatures and extremes of low foodlevels; (b) a group with need of a high levelof food, usually or often organic, and a more

limited range of temperature tolerance: ingeneral, a group of forms which multiplyrapidly under extremely favorable condi-tions, but of poor resistance; and (c) a widegroup of forms intermediate between these.The first group tends to be of very wide oc-

currence throughout the Ohio basin, whilethe second is less commonly found, and thenonly during the warm sulmmer months.Five zones have been characterized, in

terms of organic content, pH, dissolved oxy-gen, protozoan and algal plankton, and fishpopulation, ranging from grossly pollutedregions to those which are essentially sterile

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and barren. On this basis, the distributionof forms found becomes understandable.

DISSOCIATnON OF BACILLUS ALBOLACTIS.

F. S. Orcutt, Virginia Polytechnic In-stitute.Bacillus albolactis may dissociate into

smooth and rough cultural forms, whichmorphologically and physiologically are

identical. As with other cases of S and Rforms the rough is the more stable. Neitherform, however, is stable enough to give onlypure smooth or rough colonies. On stand-ard media plating of the rough form yieldsabout three per cent smooth type colonies,while the smooth form yields about seven

per cent of the rough form. Some varia-tions in environmental factors may changemarkedly the ratio of the S and R formswhen plated. Both forms of this organismappear normally in dairy products of Vir-ginia and could easily be mistaken for twodistinct species of Bacillus.

COMPARISON OF TOTAL BACTERIAL PLATECOUNTS OF SOILS ACCURATE ONLY AFTER

DEFLOCCULATION OF SAMPLES. F. S.Orcutt and A. B. Stuart, Virginia Poly-technic InstituteDilution of soil samples for plating usu-

ally involves uniform shaking to attemptequal liberation of organisms from varioussamples of soil. Retention of bacteria insoil is dependent directly upon the state offlocculation of the soil. This in turn is de-pendent upon the amount of base exchangematerial in the soil and the type of basesassociated with it.

It follows that widely different soils andsoils of the same type under variable treat-ment may be in various states of floccula-tion, so that plate counts made upon themare not comparable. Comparison can bemade only when all samples are fully de-flocculated to obtain uniform liberation ofbacteria in the soil aggregates.

THE POSSIBILITY OF FALSE INTERPRETA-TIONS IN TESTS FOR ACID PRODUCTIONFROM CARBOHYDRATES DUE TO PEPTONEDECOMPOSITION. F. S. Orcutt and L. A.Nutting, Virginia Polytechnic Institute.

Peptone is commonly used as the source ofnitrogen when a Bacillus is tested for pos-

sible ability to decompose carbohydrates.Acid production is taken as the usual evi-dence of carbohydrate dissimilation. Anumber of bacilli, however, also produceacid from peptone to the extent of pH 6.0.A lower pH than this cannot be taken as a

criterion of acid production from carbohy-drates because acid may be produced fromsugars without taking the pH lower than6.0. Control tubes of peptone without thesugars are necessary to make the proper in-terpretation.Ammonification from peptone concurrent

with acid production tends to keep the pHnear neutrality even though the carbohy-drate is decomposed. This may be avoidedby early observations, because alkali pro-

duction is not extensive enough at first toneutralize acid produced from sugars. Inmany cases observed there appears to be no

sparing action of carbohydrates on the pep-

tone decomposition.

PHYSIOLOGICAL DIFFERENTIATION OF MEM-BERS OF THE BACILLUS MESENTERICUSGROUP. W. B. Coffee and F. S. Orcutt,Virginia Polytechnic InstituteIn the latest edition of Bergey's manual,

the following organisms are listed as vari-ants of Bacillus mesentericus; Bacillus ater-rimus, Bacillus globigii, Bacillus niger andBacillus vulgatus. The characteristics bywhich they are distinguished from Bacillusmesentericus are not considered sufficient toretain them as individual species althoughthese characteristics include pigment pro-duction and several carbohydrate fer-mentations. In actual trial the differ-entiation given in Bergey's manual has notbeen found to be reliable. A number ofadditional physiological tests have beenmade to better differentiate the membersof this group.

COMPARISON OF METHODS FOR THE DETEC-TION OF COLIFORM BACTERIA IN MILK.L. A. Sandholzer, A. Walker and M. Strong,U. S. Public Health Service, Norfolk,Virginia.

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PROCEEDINGS OF LOCAL BRANCHES

WASHINGTON BRANCH

GEORGETOWN UNIVERSITY SCHOOL OF MEDICINE, WASHINGTON, D. C., MARCH 24, 1942

INFLUENCE OF CULTURE MEDIA AND Hy-DROGEN-ION CONCENTRATION ON PRO-

DUCTION OF COLOR VARIANCE IN CERTAINPLANT BACTERIA. Agnes J. Quirk, Bur-eau of Plant Industry, Beltsville, Md.

INFLUENCE OF RIBOFLAVIN OR THIAMINEDEFICIENCY ON FATAL EXPERIMENTAL

PNEUMOCOCCAL INFECTION IN WHITEMICE. J. G. Wooley and W. H. Sebrell,National Institute of Health.

Swiss mice fed a purified diet containingall of the synthetic vitamins that are known

to be essential for growth were more re-

sistant to a fatal infection with pneumo-

cocci type I, when inoculated by the in-

tranasal route, than were mice fed on the

same diet except for restricted amounts of

riboflavin or thiamine. These results were

obtained whether the mice were fed ad lib.

amounts of diet, if the amounts eaten by

those on the control diet were restricted to

the amounts eaten by the mice fed the de-

ficient diets, or if litter mates were pair fed.

When 5 times the amount of riboflavin in

the control diet was administered daily

either orally or by subcutaneous injections

to mice that had been fed restricted

amounts of this vitamin, beginning at the

time of exposure to infection, there was no

reduction in mortality. When 10 times the

amount of riboflavin in the control diet was

administered daily by subcutaneous injec-

tions to 30 mice fed a diet restricted in ribo-

flavin or the same amount of thiamine to

30 mice that had received the thiamine re-

stricted diet, beginning at the time of inocu-

lation with pneumococci, there was an in-

crease in the number of deaths.

ENCEPHALITIS. James P. Leake, NationalInstitute of Health.

GEORGETOWN UNIVERSITY SCHOOL OF MEDICINE, WASHINGTON, D. C., APRIL 28, 1942

THE BACTERIOLOGICAL WORK OF JOHNTYNDALL. Morris C. Leikind, Library ofCongress, Washington, D.C.

PARALYSIS OF THE IMMUNOLOGICAL SYSTEMAGAINST PNEUMOCOCCUS INFECTIONS.

Lloyd D. Felton, National Institute ofHealth, Bethesda, Md.

RECLAMATION OF USED AGAR. Howard I.Thaller, Animal Disease Station, Belts-ville, Md.

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