In our previous papers, diagnostic usefulness of bacterial agglutination test and indirect haemagglutination test test of Shigella sonnei and Shigella flexnerii was reported. To our knowledge, only a few papers described the differences in specific antibody leveles in the sera of dysentery patients and carriers in terms of three immunoglobulin classes.
This paper reported the results of serological investigations performed on 346 serum samples from 136 cases who were treated at Osaka Municipal Momoyama Hospital in 1972. These included 40 of S. sonnei-positive, 14 of S. flexneri 2a-positive, 1 of S. flexneri var. X-positive, 9 of Salmonella-positive, 5 of V. parahaemolyticus-positive, 4 of enteropathogenic E. coli-positive and 63 of entero-pathogenic organism negative cases. Serological methods used were bacterial agglutination test and indirect radioimmunoassay. The levels of individual immunoglobulin classes as respecting specific antibodies in these sera were calculated from the differences in the amounts of serum IgG, IgM and IgA adsorbed on the surface of S. sonnei organisms using indirect radioimmunoassay.
The results are as follows:
1. Bacterial agglutination test.
a) In case of S. sonnei-positive patients, significant rise of antibody level was first observed on the 6th day of illness and, after expornential increase, the level reached maximum (1: 320) around the 8-10th day. The maximum level continued until the 14-16th day and then the level declined to 1: 80 around the 3rd week. This trend is accordance with the results obtained by indirect haemagglutination test.
b) In case of S. flexneri-positive or Shigella-negative patients, only 2.1% of sera exhibited antibody level of ≥1: 20.
c) In case of S. sonnei-carriers, 66.7% of sera tested had antibody level of ≥1: 20. These results lead to the conclusion that the critical level of serum antibody for the diagnosis of bacillary dysentery by bacterial agglutination test is 1: 20.
2. Indirect Radioimmunoassay.
a) By indirect radioimmunoassay, the critical amounts of specific serum antibody against S. sonnei were 21.0, 2.0, and 1.4mg/dl, respectively for IgG, IgM and IgA. Levels equal to or more than these values were regarded as positive (P<0.001).
b) Before the 5th day of illness, the ratios of cases with positive antibody level among S. sonneipositive patients were 66.7% (8/12), 16.7% (2/12) and 16.7% (2/12), respectively for IgM, IgG and IgA. During this period, there were no cases in which positive serum antibody level was proved by bacterial agglutination test. By this fact it can be deduced that, in Sonne-dysentery, IgM antibody is detected first and that detection of IgM antibody has a diagnostic value in early stage of the illness.
c) This initial antibody level persisted until the 36th day in case of Sonne-dysentery patients. But in case of Sonne-carriers, there were no sera with detectable IgM antibody, whereas 54.5% and 36.4% of the sera showed positive level of antibody, respectively for IgG and IgA.
d) Considering only IgM and IgG, after the 11 th day, all the sera from patients possessed detectable IgM and IgG antibodies. Until the fifth day, sera with only IgM antibody were 50% of the total but between the 16th and the 20th day the ratio was reduced to 10%. On the other hand, the number of sera with both IgG and IgM antibodies increased rapidly in such a manner as 16.7% (the 1st to the 5th day), 52.6% (the 6th to the 10th day), 66.7% (the 11th to the 15th day) and 88.9% (the 16th to the 20th day). Sera with only IgG antibody were not observed throughout the course of illness.
e) Considering three immunoglobulin classes, in case of Sonne-dysentery patients, sera without detectable amount of antibodies were 25% of total cases until the 5th day, but after the 6th day all the sera contained antibodies of at least one immunoglobulin class and the antibody titer increased rapidly up to the 10th day.

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The fecal bacterial flora and the sensitivity of E. coli and Klebsiella to currently used antibiotics were studied in 140 infants consisting of 55 patients of acute diarrhea without underlying disease, 14 patients with underlying disease and 70 non-diarrheal healthy infants. The drug resistant bacteria of E. coli and Klebsiella were examined for the retention of transmissible resistance factors, (R-factor was detected by using K-12 strain 1310) and the following results were obtained.
1) The fecal flora of the patients of diarrhea more frequently contained Klebsiella (66.7% and Staphylococcus aureus (60.9%) than that of healthy infants contained Klebsiella (40.1%) and Staph. aur. (46.5%) (p<0.01)
2) The fecal flora of the patients of acute diarrhea with underlying disease more frequently contained Proteus (71.4%), Pseudomonas (85.7%) and Candida (35.7%) than did the patients without underlying disease or healthy infants. (p<0.01)
3) There was scarecely significant difference in the antibiotic sensitivity of fecal E. coli and Klebsiella between the patients and healthy infants.
4) From the view point of resistant pattern to Sulfonamide, Streptomycin, Chloramphenical and Tetracycline, most of E. coli and Klebsiella showed the resistance to these drugs. 48.4% of E. coli and 42.0% of Klebsiella in the patients had resistance to the four drugs concurrently. This percentage was significantly higher than that of healthy infants, namely 38.5% in E. coli and 28.0% in Klebsiella.
5) Two or three different resistant patterns of E. coli or Klebsiella were easily found in the feces of the patients and healthy infants.
6) R-factors were demonstrated in 41.4% of E. coli and 30.4% of Klebsiella isolated from the patients. This was a higher proportion compared to 35.5% of E. coli and 15.5% of Klebsiella from the healthy infants.

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The changing aspects of fecal bacterial flora and the drug resistance of E. coli and Klebsiella were examined in the patients of acute infantile diarrhea treated with various antibiotics. The culture of of feces was carried out four times, i. e., before and during administration of the drug, and immediately after and one week after ceasing medication. The drugs administrated were Tetracycline (TC), Chloramphenicol (CP), Nalidixic Acid (NA), Kanamycin (KM), Ampicillin (AB-PC), Cephalexin (CEX), Cephradine (CED), and Rifampicin (RF).
The following results were obtained.
1) There observed no notable changes in the total number of bacteria by NA, presumably because the Enterococci were scarcely affected by the drug. Regarding E. coli, a decrease of about 1/1000 was observed in more than half of the cases tested. The transient aquisition of resistance to NA in E. coli was observed during the treatment with NA in some cases.
2) When KM was administrated, E. coli decreased remarkably. In all cases, they disappeared almost completely during the administration. A week after administration, however, it recovered to the same level as before the treatment. Klebsiella disappeared after the KM administration in some cases. In cases which failed to show changes in the total number of bacteria, there used to be only minor changes in enterococcal flora.
3) When AB-PC was administrated, the decrease in the total number of bacteria was observed in many cases. E. coli decreased during the administration to the extent of being undiscernible in 6 cases, but did not decrease so much in 3 cases in which E. coli showed transient resistance to AB-PC. The decrease of Klebsiella was not so much as that of E. coli because of the natural resistance of Klebsiella to AB-PC.
4) There scarecely observed changes in the bacterial flora and resistance of bacteria following administration of CEX.
5) When CED was administered, E. coli was markedly decreased, but in one case it increased during the treatment. E. coli in this case, however, did not show resistance to CED. Any other bacteria was not affected by CED.
6) When RF was administered, there observed no changes to the bacterial flora. E. coli and Staph. aur. acquired high resistance to RF after 24 hours of the administration in all cases. This newly acquired resistance of E. coli continued for 3. months in one case.
7) The changes were scarcely observed in bacterial flora by CP or TC. I think that antibiotics must be used in infection of intestine under the consideration of its absorption and excretion, resistance of bacteria to the drugs, and its influence to bacterial flora.

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