Döderlein bacillus. Cultural and serologic studies

DtiI)HHT,EIN

HAClll,l,lIS.

(ILTlYl’IIKAI,

ANI)

SEHOliO~;l(t

STUDIES” (CLAUDE

P.

BROWN,

M.D.,

AKL)

JSDWARD

HXDOWITZ,

PHILADELPHIA,

PA.

I

N 1892 DGderleinl announced the existence of a gram-positive bacillus in the vaginal tract of normal pregnant women, which he believed to be a natural inhabitant and considered beneficial to the host. Although a considerable amount of work has been done in the study of the Dijderlein bacillus, on the prevalence, isolation, cultivation on the various media., etc.; the question as to its classification still remains an open one, and it has not as yet found a place in Bergey’s Manual. In some of the most recent papers attempts have been made to classify it with L,. otio@hilus. Lucy M. Brycea in a study of 119 eases of pregnant women, found what she called DSderlein bacillus in smears of 84 cases; however, it was cultured in only 62 eases. Using ten strains in her studies with carbohydrates, she states “the ob served characteristics agree with those of the genus Lactobaeillus as described in Bergey ‘s Manual. ’ ’ However, the strains varied in their carbohydrate fermentation and in conclusion she states that “I;. acidophilus is a frequent inhabitant of the intestinal tract, therefore, it is easily conceivable that during infancy or childhood, organisms of this type may gain entrance to the vagina which is sterile at birth and finding there a particularly suitable environment establish themselves as the predominating species.” ThornaG made comparative studies of DSderlein bacillus with L. c~&do~hhiEzcs and L. bulgan-ium. In his study, eight strains of Xderlein bacillus isolated from normal children six to twelve years of age were used with two control strains of each of L. acidophilus and L. bulg&czls. In most cases isolations were made from anaerobic plates and in one case from an aerobic plate. The fermentation reactions reported were irregular, and there was no clear-cut difference between DGderlein, Acidophilus and Bulgaricus strains. In his agglutination tests none of the sera prepared from DGderlein strains agglutinated L. butgarricn*s antigen, 6 out of 8 agglutinated the acidophilus antigen, however the DBderlein sera agglutinated only one to five of the heterologous Diiderlein antigens. Thomas re-isolated every strain of Diiderlein from the feces of human beings, after feeding them with the culture in milk, and draws the conclusion that DKderl&i’s vaginal bacillus is L. aaidophil?Ls and that this organism is present in less than 10 per cent of normal children. It gets into the vagina by exterior passage from the intestinal tract, and can be introduced into the vagina by feeding a culture by mouth. Lash and Kaplan4 studied the vagina,1 secretions in 98 pregnant women and found Diiderlein’s bacillus in smears in 41 cases but cultivated it in only 13 cases, using both aerobic and anaerobic methods. They believe their failure to cultivate the organism in a great number of (sases was due to more rapid growth of the associated organisms such as staphylococci, diphtheroids, streptococci, and B. ooli which they also found present. *Read at the meeting Racteriologists, Philadelphia.

of the Pa.

Eastern May

74,

Pennsylvania 1932.

705

Chapter

Society

of .4merican

706

AMERICAN

.JOCRNdI,

OE’

DBSTETRI(‘S

ASD

GYNECOI,OGY

They were unable to classify DGderlein bacillus by the carbohydrate fermentations, agglutination, precipitation, They also inand complement fixation tests. jected animals using large doses of forty-right-hour dextroscb broth cultures, and found none of their Diiderlein strains pathogenic for rabbits, guinea pigs, and mitt. In their opinion the term “ I~Sderlein bacillus ” includes a large group of organism8 which, though related have some differentiating characteristil,s. This fact makes it difficult to classify them and t.hep suggest a specific name, ‘( liactol~ac:3hls vaginae, ’ ’ to a member of this group whicsh differed in some characteristics froul the other strains. We agree with Thomass that this would IX, unjustifiable due to lack of any definite criterion for ciassification. Mohler and Browns in a recent paper deal with the question of ‘LD?iderleinls bacillus in the treatment of vaginitis. ” @or present studies of the Diiderlein bacillus have been made possible t.hrough the courtesy and interest of Dr. Roy W. Mohler, Associate in Gynecology at, Jefferson Hospital, who obtained the cultures from cases in his practice. We have two strains of Diiderlein bacillus under study at t,he present time designated as Strains Ml and M2. Strain Ml was isolated from a normal virgina.l vagina on aerobic whey agar plates. It is a nonmotile gram-positive bacillus. Tt appear singly and a&o in short and long chains. Either straight or curved rods can be seen, with flat or rounded ends. Its average size is 0.5 to 1.0 p wide and 3.0 to 5.0 p long; however, some very short and very long forms can be seen depending on the age of the culture and its environment. Optimum temperature was 37” C. When first isolated, the organism grew very poorly in whey and on whey sgar plates and had to be transplanted daily for t,wo weeks to Once established it grows luxacclimate it to its new environment. uriantly in whey, having a P,, 7.6. For all routine work cultures are carried in this medium. Stock cultures were kept in milk on account of its buffer qualities which arc essent,ial for the maintenance of cultures of the aciduric group. Surface iimbriate. DFB~

Colony

on Whey

C&my.-Resembles

A.qar.----Small a rerp

tiny

filamentous, pledget

irregular,

of cotton

or the

slightly so-called

iridescent, Type

Y

r~ololly.

In JfzZlz.---Coagulation in five days, soft coagulum, no gas. Litnam Af$lL.-Turned acid in three days, coagulated in five days, no gas, soft coagulum. observation for ten says. Get&n.-No noticeable growth at 20” or 37” C., under Dextrose Broth ia Fermentation Tube.-Grew very poorly, and settled at the bottom in a precipitous sediment, no gas. It&OZ.--Not formed. Reduction of Nilrates-Nitrates reduced to nitrit,es.

Strain M2 was obtained from a married woman in whom the vaginal tract appeared normal. It corresponds in all respects to Strain Ml except for the fact that it grows much more slowly, in milk and whey.

ISROWh-

A,“*‘D

In the third culture be Dijderlein’s bacillus sequent, transplants.

REDOWITZ

:

DijDERLEIN

HA(‘II,I,I’S

from a normal case showing on the first, plate, we failed

70-i

what appeared to to grow it in sub-

In the fermentation tests we selected only those ca.rbohydrates which gave the most consist,ent results, as reported by Lash and Kaplan, Brycc and Thomas. For controls we used t,hree known strains of L. tlcidophilus, one being our own and the other two obtained on the market from well-known aeidophilus preparabions. Hiss serum-wat,er medium containing axolitmiu and 1 per cent of carbohydrate, was used. The cultures were grown in whey for eighteen hours, centrifuged, the acid supernatant discarded and sterile salt solution added to t,he organisms t,o make a heavy suspension; two to three drops of this RUSpension were added to each carbohydrate tube containing 1 cc. of medium and incubated at 37” C. No acidifying of medium occurred befo’re the seventh day and on t,he tenth day there was a complete decoloration and in most instances coagulation, but no gas. TABLE

STRAIN

DXerlein Diiderlein Acidophilus Acidophilus Aridophilus *AC,

Ml M2

1.

CARBOHYDRATE

MALTOSE

GLUPOSE

SIT(‘ROSE

LACTOSE

RAP FISOSE

MANNITF,

AC AC AC

AC AC AC

AC AC AC AC AC

AC AC SA SA AC

0 0 0 0 0

slightly

acid;

U

AC’” AC A(?

L

A(:

IC

AC

S

AC

AC

AC

acid

and

REACTIONS

coagulation

; -4, acid;

Sh,

0,

no

SALICIN --

INULIN

AC 0 AC A AC

0 0 0 0 0

change.

Agglutinating sera were prepared from all the strains used. The Diiderlein strains did not agglutinate each other nor any of the acidophilus stra.ins. None of the acidophilus sera agglutinated the DGderlein antigens. Two strains of acidophilus isolated from milk showed cross agglutination. These results of the agglutination tests werr checked by absorption experiments. TABLE

11.

AGGLIJTINATION

RF,AOTIOI\‘S

ANTIGENS

ANTISERA

Diiderkin Ml DGderlein M2 Acidophilus U Acidophilus L Acidophilus S *0 = negative.

DijDERLEIN Ml

+” n 0

n ?u

U

0

0 + = positive

ACIDOPHILUS U

DiiDERLEIN ,112

0 (agglutination).

0 0 t

ACIDOI’HILUS I,

0 0 0

ACIDOPHILUS S

0 0 0 +

1:

f+

+

708

AMERICAN

,JOT?RNAI>

OF

OH8TETRICG

AND

GYNECOI,OC,Y

The two strains of T~iidrrleiu haeill i absorbed l-he agglut,initts from the homologous but not, the hetcrologous serum. Of t,he three strains of acidophilus which WPI’C tcstrtl, each one absorbed it,s homologous agglutinin. Strain U did not absorb the heterologous agglutinins, however, Strains L and S absorbed the agglutinins produced by each other but not of U strain or the two strsins of Diiderlein bacilli.

CONCLTJSIONS

Two strains of Daderlein’s bacillus were isolated from normal vaginal secretions on aerobic whey agar plates and compared with three strains Df L. acitiphilus. The colony and cultural characteristics of DGderlein bacillus and C. acidophilus are very similar and the fermentation reactions interlock sufficiently so that differentiation cannot be made on those grounds. Dijderlein bacillus is serologically different from L. uei&phdus, in fact the cult,ures under study differed from each other. REFERENCES

( I ) Dtiaertein. : Verhandl. d. deutsch. Gesellseh. f. Gyniik., 1892. (2) Br?lce, Luoy Y. : Med. J. Australia 1: 793, 1928. (3) Thomc, Run&y: J. Infect. Dis. 43: 218, 1928. (4) Lash and Eaplm: J. Infect. Dis. 38: 333, 1926. (5) iKo&lm, IlO!/ JV.> am2 Claude P. Brown: Ax. .T. ‘@ST. & GPNEC. 25: 718, 1933. 1930

CHEWNUT

STREET

EXPERIMENTAL

LIGAMENTOUS GUINEA

W. T. ‘From the Lkpartmmd

PIG M.D.,

POMM~WKE,

of

Ohstetrits

am3

RELAXATION

IN

THE

PELVIS ROUHWTER,

Gynecokqy,

N. Y. tha Un6vewitg

of

Rochester)

ORPHOLOGIC changes of the pelvis in a number of species of mammals approaching sexual maturity and during pregnancy lave long been recognized. Although in a number of animals such :hanges are associated with actual bone absorption, as in the pocket gopher (Hisawll) , these alterations ordinarily involve the inte&ubie !igaments at the symphysis and those at the sacroiliac arti-icufation~ so

M