The study was carried out for the purpose of enhancing the detection rate of enteral pathogens in diarrheal patients. The stool samples were obtained from 140 inpatients at The Kobe Communicable Disease Hospital and 58 outpatients at The Kobe Municipal Central Hospital, and they were cultured by various methods on various culture media. As control, samples were obtained from 1, 013 healthy people, and the same media and culture methods mentioned above were applied to them.
1. Enteropathogens were detected in 32.8% of the patients with diarrhea, of which 12.6% was Salmonella, 10.6%, Enteropathogenic Escherichia coli, 5.1%, Shigella, 3.5%, Vibrio parahaemolyticus and 1.0%, Plesiomonas shigelloides. No case of diarrhea due to Staphylococcus aureus, Clostridium perfringens or Yercinia enterocolitica was observed or suspected.
2. Enteropathogenic Escherichia coli, Salmonella and Shigella were detected in the healthy people at the rate of 3.2%, 0.2% and 0.1%, respectively.
3. For the detection of enteropathogenic Escherichia coli in the patients with diarrhea, the culture of diluted stool specimens was better than the direct culture.
4. The viable counts of aerobes in the patients stools were within the range of 0-¹⁰⁹/g (ml). In 19.2% of the samples, the viable counts of aerobes were less than ¹⁰⁴/g (ml).

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Three patients with subacute bacterial endocarditis were treated with large doses of penicillin G and cephalosporin antibiotics. Minimum inhibitory concentrations of penicillin G for streptococci isolated from three patients were 0.1mcg/ml or less. Those of cephaloridine were 0.1mcg/ml for 2 strains from case 1 and 2, and 0.5mcg/ml for the other one from case 3, respectively.
In case 1 infected with streptococcus lactis, the treatment with 18 million units of penicillin G per day failed to cure and daily 14 g of cephalothin combined with 6 g of cephaloridine caused renal impairment and hypersensitivity phenomena, i.e. high fever, thrombophlebitis in the site of intravenous drip, exanthema, liver enlargement, elevation of GPT, and leukemoid reaction. Cure was achieved by the daily use of 28.8 million units of penicillin G, and sodium polystylene sulfonate (Kayexalate ^®) was effective on hyperpotassemia of 7.6 meq/L caused by large doses of potassium penicillin G in the circumstance of renal impairment. Total dosis of penicillin G used was 2805 million units.
Case 2 infected with streptococcus dysgalactiae was treated with 6 g per day of cephaloridine and cured without any side effect. Total dosis of cephaloridine used was 195 g. In case 3 infected with streptococcus mitis, treatments with 6 g per day of cephaloridine and 29 million units of penicillin G plus 2 g of streptomycin in every day failed to cure, respectively. The infection was suppressed by the dosis of 44 million units per day of penicillin G but symptoms of aortic insufficiency were remained after the completion of chemotherapy. Total dosis of penicillin G used was 2470 million units but no hyperpotassemia appeared.
Another phenomenon observed in each patient was appearance of broad albumin band on the serum electrophoresis. This seemed to be due to binding of penicillin G and cephalosporin antibioitcs to serum albumin. No coomb's positive reaction was observed.

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Serum antibiotic level is regarded as an important guide, especially in endocarditis or other bacteremic diseases, for predicting antibiotic effects and making subsequent therapeutic programs.
As for the evaluation of serum antibiotic level from the clinical view point or the methodology of serum antibiotic assay itself, however, there have not necessarily been standardized lines yet.
In this paper, the author scrutinized the interrelationship between estimations of serum antibiotic level measured by various methods and clinical effectiveness using four bacteremic patients, the causatives being Staph. epidermidis and Staph. aureus in two sepsis cases (case 1, case 2), respectively, Strept. lactis and Strept. disgalactiae in two endocarditis cases (case 3, case 4), respectively.
The summary is as follows:
In Cup Method using Bacillus subtilis as an indicator, the serum antibiotic concentrations were shown six times or more higher than MIC. In spite of this fact, clinical improvements had hardly been achieved. This method cannot be considered adequate for therapeutic guide.
However, antibiotic containing serum inhibitory power to the isolated agents from the patient (case 3) measured by Cup Method using the isolates as an indicator has been consistent with clinical results.
Antibiotic containing patient sera were tested for growth inhibitory effects to the isolates from the patients by Tube Dilution Method. In case 1, the sera diluted to 1: 2 showed inhibitory effects, but the treatments were proved fruitless clinically. In the other three cases, the treatment succeeded when the sera diluted to 1: 8, 1: 32 and 1: 128, respectively, inhibited the growth of the isolates. In case 3, however, the treatments were unsuccessful while the serum bacteriostatic titer remained 1: 32 under daily 18 million units PC-G regime. To the author's impression, antibiotic therapy based on serum bacteriostatic activity may fail in such a case as the MIC of patients is much lower than its minimal bactericidal concentration.
Bactericidal activities of patients' sera agaist causative organism were assayed by the method described by Schlichter et al. The decline of bactericidal titer of PC-G in the sera to Strept. lactis isolated from SBE case (case 3) was observed when the large dose of organism was inoculated to the medium. Despite daily 18 million units PC-G plus daily 8 gr Lincomycin were administrated in that case, a small number of colonies remained on the medium resisting low or non diluted patient's sera, when undiluted freshly cultured broth was employed. At the same time, the therapy resulted in failure clinically under that dose. The patient was finally cured under the long term treatment of daily 28.8 million units PC-G, however. And, in the case of undiluted freshly cultured broth, 1: 8 dilution of the serum, and in the case of 1: 100 dilution broth, 1: 128 dilution of the serum completely killed the pathogens on the media, respectively. In case 4 of SBE (Strept. dysgalactiae), who was failure clinically under that dose. The patient was finally cured under the long term treatment of under daily 6 gr Cephaloridine regime, 1: 32 dilution of the serum completely killed the causatives even when undiluted fresh culture broth was inoculated. MIC of Cephaloridine was, then, disclosed as 0.1mcg/ml. In this patient, Cephaloridine was conspicuously effective.
It is the author‘s viewpoint that in most cases of bacteremic patients who are poor in the natural healing tendency, most conventionally used bacterial inhibition tests cannot be regarded as adequate guide for antibiotic therapy. From the author’ s experience, undiluted culture broth should be included as the test material among others.

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Each strain of E. coli, Klebsiella, Enterobacter, Serratia and Pseudomonas, isolated from clinical materials in recent years was cultured in heart infusion broth, and the broth culture was sterilized with membrane filters. Then, the amounts of PC-G, AB-PC, CB-PC, SB-PC, MCI-PC, CET, CER, CEX and CEZ inactivated with 1 ml of the filtrate were examined. The antibiotics of PC group tended to be inactivated by the filtrate in the following order: PC-G<AB-PC<CB-PC=SB-PC<MCI-PC. The antibiotics of Cephalosporin group were as follows: CER<CET=CEZ<CEX.
The measurement of amounts of AB-PC and CET inactivated with 1 ml of the filtrate after 48 hours culture of various kinds of GNR, showed that most Klebsiella strains mainly inactivated AB-PC, and that most Serratia and Pseudomonas strains inactivated CET. About a half of the Enterobacter strains inactivated CET and the other half of them inactivated both AB-PC and CET. Among E. coli, Klebsiella and Enterobacter strains, the strains which inactivated-both AB-PC and CET showed extremely high values of inactivating amounts; many strains had the ability to inactivate several ten thousands to 300, 000μg. It was confirmed that the inactivating ability of the antibiotics was demonstrated to be accumulated in the broth culture with time.
“Double Disc Method”, a simplifiedbiological method for assay of β-lactam antibiotic inactivating substance of GNR, was developed in our laboratory. In this experiment, a close relationship was observed between the results with this method and the antibiotic inactivating ability produced in the bacterial culture filtrate, and its availability of clinical application was suggested. In addition, it was considered that β-lactam antibiotic inactivating substance in this experiment was attributable mainly to β-lactamase.

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[in Japanese]

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