EXPERIMENT STATION BULLETINS. 409 



but we are confident that it is a valuiible measure worthy of trial pend 

 ing a fuller understanding of <lie nahire of the conditions inv^olved. 

 Our experience wilh this trealnient warrants us in leconimending it as 

 superior to other methods of treatment in all cases of retained i)lacenta 

 and following all abortions. Work for two years on the bacterial flora 

 of the vagina by Mr. Knopf and Mr. nimmelberger does not indicate 

 that the lactic organism establishes itself in the vaginal mucosii and 

 that an undue acidity of the mucous membrane is not produced. We 

 hope to continue this investigation of the bacterial flora under various 

 conditions. We are making a report on our work with the abortion 

 organism although it is incomplete and inconclusive in many respects. 

 We have used three cultures that have the characteristics of the Bang 

 organism, viz. (1) '^Goda" isolated by us May 1st, 1911 from cotyledon 

 of placenta from Guernsey cow. Both foetus and placenta were removed 

 and cultures made immediately on serum-agar-gelatine. Three out of 

 live tubes gave typical pure cultures; (2) "MacNeal" sent by Dr. MacNeal 

 from Illinois, and (3) "Good" from Prof. Good of Kentucky. 



In general, it may be said that growth with these organisms, especi- 

 ally with "Goda," is not vigorous until after many transfers in vitro. 

 We wished to obtain the most suitable medium for mass cultures. 

 During this attempt, we made use of the pregnant uterine wall, the 

 foetal membranes, the foetus and the amniotic fluid separate!}'. In 

 case of the three substances first mentioned, liquid and solid media were 

 made as is usual with fresh chopped meat. The reaction was not cor- 

 rected in any case and was found to be rather acid in case of uterine 

 wall. None of the media made in this manner was more successful than 

 the usual serum-agar-gelatin, that from the uterine wall was useless 

 possibly on account of acidity. The amniotic fluid was used as a sub- 

 stitute for bouillon after filtering and sterilizing by discontinuous 

 method without correcting reaction or addition of any nutrients. Addi- 

 tion of 1.5 per cent agar made a very satisfactory solid medium. The 

 amniotic fluid media gave better results than any media yet tried. The 

 reaction was neutral or slightly alkaline. 



The following experiments with virgin heifers were conducted as pre- 

 liminary efforts toward the production of immunity by the use of liv- 

 ing cultures of Bact. abortus injected subcutaneously. We adopted this 

 method assuming both that the subcutaneous injection would result in 

 stimulating the production of suitable antibodies antagonistic to future 

 invading organisms of the same species and also that the organisms 

 would disappear from the system without contaminating tlie environ- 

 ment. 



May 1, 1911, inoculated with "MacNeal" culture flasks containing 

 30 c.c. of 5 per cent glycerine bouillon -f 10 c.c. of naturally sterile 

 horse serum. Flasks sealed with paraffin, clouded in about four days 

 and increased in cloudiness after which sediment fonned. Usually, after 

 a month the plug is drawn into the neck of the flask, sometimes entirely 

 within. 



Two virgin Jersey heifers were received June 3, 1911. One is referred 

 to as Light, the other Dark. Their previous history not known. 



June 8, drew blood from jugular of each. Mucosa of vulva of Dark 

 was smooth and pink, that of Light was congested and very rough with 

 papules. 



