In May 1979, a strain of Serratia marcescens was cultured from arterial and venous blood, sputum and urine of a patient who was admitted in the Iwaki Kyoritsu General Hospital. The strain showed low susceptibilities to almost all commercially available parenteral antibiotics (over than 100 mg/ml) except for gentamicin, amikacin and minocycline (50 mg/ml). The disease of the patient was diagnosed as septicemia due to multiple resistant Serratia marcescens and he was moved to an isolation ward. Soon after isolation of this patient, Serratia marcescens showing the same susceptibility pattern was cultured from the urine of another patient at the same ward who used the same bed that had been used by the isolated patient. At that time a possibility of a wide intrahospital spread of an opportunistic pathogen was suspected and thus retrospective study of microbiological laboratory records was conducted. It was found that emergence of infection caused by the epidemic strain could be traced back to November 1978, i.e. the epidemic strain had already cultured been from a patient who had died in November 1978 from uremia associated with severe urinary tract infection. Thus it was suspected that the epidemic spread originated in the urological ward. A retro- and pro-spective epidemiological and microbiological survey of prevalence of the epidemic strain was conducted from October 1978 through September 1979. The incidence increased from November 1978 to July 1979, when intensive control measures against the spread were instituted. Thereafter, the incidence became lower. During the period, from November 1978 to July 1979 a total of 132 patients were infected with the epidemic strain. The highest incidence was seen in the urological ward followed by that in the coronary care unit and that in the ward of obsterics and gynecology. Altogather 126 of the epidemic strains were isolated by October 1979. Most of them, 113 strains, were recovered from urine specimens and 5 were from the sputum. Three strains each were obtained from blood and pus of skin abscess respectively. One strain each was cultured from bile and an endometrial discharge. Thus it was known that the site of colonization of the epidemic strain was confined almost exclusively to urinary tract.
A committee of doctors, nursing and paramedical staffs was organized and an active educational campaign against intrahospital spread of the epidemic strains of Serratia was started. This compaign, with the other intensive control measures, succeeded in decreasing the incidence of infection due to the epidemic strains.
From the results of bacteriological examinations, the epidemic strain was shown to be distinctively different from the background strains of Serratia marcescens in the hospital. Twenty strains of Serratia marcescens isolated in the hospital were selected at random and the serological typing of them was made. All of the 16 strains, which showed the same antibiotic susceptibilities as those of the epidemic strain were grouped into serotype 04. Out of 4 strains, which showed a different susceptibility pattern from the epidemic strain, 2 belonged to serotype 04, and a strain did to serotype 01 and in a remaining strain serotype was not differentiable.

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A total of 863 cultures of staphylococci were obtained from various clinical specimens at Bacteriological Laboratory in Kyorin University Hospital, during a period from March through August, 1980. These isolates were composed of 212 of S. aureus (24.6%) and 649 cultures of coagulase-negative staphylococci (75.2%). Coagulase-negative staphylococci were subjected to species classification by criteria of Kloos & Schleifer. As a result, out of 649 isolates 90 wer e isolated from each specimen of blood, cerebro-spinal fluid, pus and ascites, and also urine with the coccal number more than 105 CFU/ml. These 90 isolates could be classified into 9 species except those of S. hyicus and S. sciuri. The results suggested that most of all coagulase-negative staphylococci have pathogenicity to humans, and that S. xylosus may be an important species of staphylococcal infections in the urinary tract, since 5 of 11 urinary isolates were classified to the species.
Although novobiocin susceptibility test have been carried out as an important characteristic for differentiation of S. saprophyticus from other species, in this survey, major coagulase-negative staphylococci; S. xylosus, S. saprophyticus and S. warneri, were found to be novobiocin resistant in the incidence of 5/16 (31.3%), 8/29 (27.6%) and 8/40 (20.0%) respectively. Accordingly, it seems unlikely that novobiocin resistance is a key characteristic to differentiate S. saprophyticus from other species.
As to sensitivity tests of major isolates to 10 antibiotics, of 103 S. aureus 21 strains showed resistant to any of the drugs tested, and 39 multi-resistant to 2 or more drugs. Among coagulase-negative strains, S. epidermidis and S. hominis were found to be especially a high incidence of multi-drug resistance, 26/36 (72.2%) and 23/32 (71.9%) respectively. Also out of 53 strains classified to the other species, 16 were multi-resistant ones.

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This study was aimed to clarify the seroepidemiological status of pregnant women against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by microneutralization test and enzyme-linked immunosorbent assay (ELISA). The following results were obtained.
1) Incidence of the neutralizing antibodies to HSV-1 and HSV-2 in 90 females in Fukuoka area was dependent on the age. In the age groupof 1to 20 years the negative antibody ratio was 70 to 80%, whereas it was less than 10% in the group of 41 to 60 years of age. Changing pattern was seen in the group of 21 to 40 years of age.
2) Pregnant women seronegative for HSV-1 and HSV-2 were found 34.1% and 37.1% in Fukuoka (n=132), 30.6% and 31.5% in Miyazaki (n=124), respectively.
3) Testing 100 sera from pregnant women, the HSV antibody titers determined by ELISA correlated very well with those by microneutralization test.
It was thus evidenced that approximately one third of pregnant women in Kyushu lacked the antibodies to HSV-1 and HSV-2. This means that those seronegative pregnant women can not endow their infants with the antibody-mediated protction against neonatal HSV infection. ELISA appears to be useful as one of the routine tests for screening the antibodies to HSV.

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The protective activity against infection (SPA activity) of the aseptic filtrate from cultures of S. enteritidis2547, a high SPA-producing strain, in TSA medium reached a plateau at 4 days of cultivation and remained at essentially the same level over the ensuing 30 days. As progressive bacterial autolysis occurred in the culture on incubation, however, it was decided to use culture fluid harvested after 4 days of incubation at 37°C and sterilized by filtration, for purification of SPA.
The filtrate of culture fluid thus obtained contained SPA activity which was stable between PH 5.0 and 9.0 and resistant to heating at 100°C for 30 minutes.
One hundred liters of the aseptic culture fluid filtrate was concentrated to one-hundredth of the original volume and purified by TEAE-Cellulose and DEAE-Sephadex A-50 column chromatographies and then by gel filtration on Sepharose 2B.
The final purified preparation proved to form single precipitation line by Ouchterlony immunodiffusion with a mouse immune serum to a crude SPA preparation. It also showed a single spot on high voltage paper electrophoresis. Qualitative analyses by color reactions disclosed the prepation to be a polysaccharide free from lipids, proteins and nucleic acids.

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In the old-aged, pneumonia is often based on organic lesions of the lung itself, and decrease in general resistance is often observed as a background. Therefore pneumonia causes a high mortality in the old-aged in spite of the development of antibiotics in recent years.
Among the patients aged over 60 years with senili pneumonia who had been treated with combination of immunoglobulin (“venilon”, a sulfated immunoglobulin preparation) plus antibiotic therapy, we selected retrospectively the cases where the results of laboratory examinations including X-ray observation of the chest and bacteriological examination of the sputum, and the clinical course had been obviously known, and we evaluated the utility of the concommitant use of immunoglobulin preparation and antibiotics in these cases. For this study, a committee was composed of 7 physicians.
The evaluation varied individually according to different members of the committee, but their opinions agreed almost unaimously for 9 cases. Although there are some different opinions concerning the efficacy of combination of immunoglobulin with antibiotics, the present study revealed the result considered “Certainly effective” in some cases.
It is said that the efficacy of immunoglobulin preparation originates from a specific antibody contained in it. However, further investigation will be necessary to clear its mechanism.

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Recovery of fecal and pharyngeal flora after total intestinal decontamination was investigated in 3 patients undergoing autologous bone-marrow transplantation (2 with malignant lymphoma and 1 with nasopharyngeal carcinoma) on the protected environment-prophylactic antibiotic program.
The oral nonabsorbable antibiotic regimen consisted of gentamicin, vancomycin and nystatin (GVN). In some instances dibekacin was substituted for gentamicin (DVN). In addition, these antibiotics were sprayed in the mouth for oropharyngeal decontamination.
Antibiotic prophylaxis with GVN was continued for 5 weeks in 2 patients and DVN for 6 weeks in the third patient. Fecal flora was completely suppressed one week after initiation of GVN. Stools remained “sterile” for the following 4 weeks. On the other hand, use of DVN produced only partial suppression of lactobacilli and candida despite complete elimination of other fecal organisms.
One week after cessation of antibiotic prophylaxis, the concentration of fecal organisms returned to the pretreatment level in all 3 patients despite the use of bioclean rooms and sterile food. Aerobic fecal flora was composed of the same spectrum of organisms as that found in pretreatment specimens. It took longer periods of time for anaerobes to reappear in posttreatment stools. Clostridia returned within one week following cessation of the oral antibiotics; bacteroides and peptostreptococci reappeared following 3 -4 weeks. Veillonellae and bifidobacteria reappeared after the intermediate period.
Recolonization of phryngeal flora, which was only partially suppressed during antibiotic prophylaxis, followed almost the same process as that observed in fecal flora. However, it took more than one month for neisseriae to reappear in the throat of 3 patients.
In conclusion, it was suggested that the reappearance pattern of fecal and pharyngeal flora after total intestinal decontamination is similar to that observed in newborn infants and it takes about one month for fecal flora and more than one month for pharyngeal flora to fully reappear.

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As one aspect of research to clarify the defence mechanism in enteric infection, comparison of phagocytosis to adherent cells from peritoneal cavity (macrophages) and infectivity to HeLa cells between Shigella flexneri 2b 17-A (a virulent strain) and Shigella flexneri 2b 17-N (an avirulent strain) was carried out. The findings of the experiments may be summarized as follows:
1) Bacillus suspensions of both strains were contacted with macrophages or HeLa cells at 37°C for 1 or 2 hours. After separating bacillus by washing, these cells were incubated further for the confirmation of their phagocytic or infection rates. Phagocytic rates of the virulent strain or the avirulent one in macrophages reached about 20% in 0-2 hours after incubation. The rate of the former increased with the lapse of incubation time but that of the latter trended to decrease. HeLa cells were infected with the virulent strain but not with the avirulent strain.
2) In the case of phagocytosis of macrophages with larger contact dose of bacteria or longer contact time, initial phagocytic rates (at the incubation on 0 hr) were increased and destractive degree of macrophages were enlarged. So, there could hardly be found any difference on an initial phagocytic rate and destructive degree of macrophages between the virulent strain and the avirulent one.
3) Macrophages were contacted by both strains treated with 70% alcohol for 30 minutes or 100°C for 30 minutes, thereafter they were incubated further. Initial phagocytic rates of them showed about 20% and then they decreased along with the incubation time. Eight hours after incubation, there were not found bacteria in any macrophage. The same experiment was carried out with HeLa cells, and bacteria were not found in HeLa cells at any time of incubation.
4) A morphological analysis of phagocytes was performed using transmission electron microscope. Immediately after inoculation of the virulent or the avirulent strain into the peritoneal cavity of guinea pigs, their phagocytes began to ingest them. In phagocytes from the guinea pigs 4 hours after the injection of the virulent strain, most of their phagocytic vacules were of a loosed type, but that in phagocytes from the animal injected with avirulent strain trended to be of a tight type.
A limiting membrane was recognized around the bacilli in phagocytes from animal inoculated with both strains, irrespective of their virulence. On the contrary, the membrane did not found around the bacilli in HeLa cells.

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Impression that L forms in blood culture will be encountered only in extreme rarity is still commonly held by workers in the field of infectious disease.
Recently in our laboratory, we were able to isolate L forms of Streptococcus bovis from blood cultures.
The patient was a 54 years old man suffered from polymyositis, and during his predonisolon therapy, high fever and leukocytosis occurred and ESR rose to 52 mm/h and CRP turned to strongly positive.
Ampicillin, Cefazoline, and Carbenicillin revealed no effects.
During this period, blood specimen were cultured three times, and we could isolate L forms, from one of these specimen which reverted into Streptococcus bovis.
We were “Culture bottle Eiken No 3” as a culture medium and the osmotic pressure of that medium was 331 mOsm.
This case suggests that antibiotics affecting cell wall synthesis may induce L forms and which may be one cause of persistent infection.

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