To develop a new animal model of intraabdominal abscess by Escherichia coli alone, we reevaluated anaerobes and other additions which had been believed necessary to produce an intraabdominal abscess. We took the method of bacterial implantation by insertion of a double gelatin capsules containing microbes and the additions into the peritoneal cavity of male Wister rats. We examined the requirement of causative bacteria for an abscess including both aerobes and anaerobes, sterilized feces, and barium sulfate. It has been proven that a simple and well reproducible intraabdominal abscess can be developed without fail at the seventh day after inoculation although anaerobic bacteria, sterilized feces, and barium sulfate are not used. However, we have failed to produce an abscess without sterilized gauze fiber which should be a core of an abscess and is used instead of sterilized feces. This animal model will contribute to a major simplification of the original one heretofore in use, and is expected to serve as an aid to elucidate the mechanisms of abscess formation.

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It has been demonstrated that an intraabdominal abscess by Escherichia coli alone can be developed without fail although anaerobes or barium sulfate are not used. We investigated the properties and the influence of this abscess on the host. We took the method of bacterial implantation by insertion of a double gelatin capsules containing Escherichia coli suspension of which concentration was adjusted to five grades into the peritoneal cavity of Wister rats. Abscesses were developed in the survived rats on which live bacteria had been inoculated. Only Escherichia coli were found in these abscesses by culture whereas no death was occurred and no abscess was developed in the rats on which no bacterium or heat-killed ones had been inoculated. As for non-survivors at the 7th postoperative day, all of them died of panperitonitis and no abscess was developed. An abscess was developed without fail when live bacteria of which number within the order of 10⁷ colony forming units were inoculated. Blood endotoxin concentration 24 hours after inoculation increased exponentially according to the inoculum size. However, that at the 7th postoperative day returned to the levels at zero time. Microscopic examination revealed a thick abscess wall, poor infiltration of inflammatory cells, and poor neovascularsis into the wall. These findings suggest that endotoxin is prevented from release into the blood stream since abscess contents are isolated by thick wall.

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Chlamydia trachomatis detection by both swab CHLAMYDIAZYME and urine sediment AMPLICOR were performed on 61 males and 54 females who were suspected to have C. trachomatis genital infection (CTGI). Positive agreement between swab CHLAMYDIAZYME and urine sediment AMPLICOR was good, 96.9% in males and 96.6% in females. Negative agreement was 100% in males and 84.0% in females. “Swab CHLAMYDIAZYME positive-Urine sediment AMPLICOR negative specimen”were not obtained. “Swab CHLAMYDIAZYME negative-urine sediment AMPLICOR positive”resulted in four females who were finaly diagnosed to have CTGI by repeated specimens on CHLAMYDIAZYME. Positive result by AMPLICOR was obtained on un-spun urine specimen that was in agreement with the laboratory result that the sensitivity of AMPLICOR reaches one thousand fold of CHLAMYDIAZYME. Detection on every separately collected urine specimen of one urination showed AMPLICOR positive all through the last cup specimen in both male and female. Those results seem to certify the reliability of C. trachomatis detection by urine sediment AMPLICOR. False negative by interference substance in urine and false positive by contamination were not experienced. On asymptomatic young male and female, positive rates of urine sediment AMPLICOR were 7.5%, in male and 12.1% in female which are far more than the positive rate obtained by CHLAMYDIAZYME. Non invasive C. trachomatis detection method applicable to asymptomatic female is needed for control of CTGI. C. trachomatis detection could be faciliated by non invasive urine sediment AMPLICOR.

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Coxsackievirus A16 (CA16) and enterovirus 71 (EV71) are known to be major causative agents of hand-foot-and-mouth disease prevalent in summer in Japan. Discrimination and identification of these viruses were oten hampered by a nonneutralizable or nontypable virus. Therefore, a Southern blot hybridization that utilizes mixed probes specific to serotype was developed. Firstly, an approximately 650 bases spanning 5'-noncoding region to one third of VP2 including entire VP4 was amplified with a set of primers containing enterovirus common sequences and a genomic RNA as template. Secondary, the nucleotide sequences were determined using seven CA16 and eighteen EV71 strains including the standard strains, and the deduced amino acid sequences of VP4 were searched to find residues which are conserved in the same serotypes but diverged among different serotypes. Candidate positions for the mixed probes were defined at the carboxyl terminus of VP4. Thirdly, Southern blot analyses were carried out using thirty-nine enterovirus standard strains, seven CA16 isolates and sixty-six EV71 isolates previously identified by the neutralization test. The results revealed that each mixed probe exclusively bound to the homologous DNAs but not to the heterologous ones.
In an attempt to determine serotypes without virus isolation, clinical specimens from hand-foot-and-mouth disease were examined. Of 78 throat swabs and 15 vesicular fluids, 71 (91.0%) and 13 (86.7%) specimens were clearly identified, indicating that the method described here offer advangates over the traditional neutralization assay: It is rapid, specific and less labor-consuming.

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Out of 217 strains of viruses isolated from patiants with conjunctivitis at one ophthalmology clinic in Nagoya City from 1982 to 1995, 37 were adenovirus type 37 (Ad37). They were isolated mainly from patients with epidemic keratoconjunctivitis in summer-time. DNAs from all Ad37 isolates were extracted and analyzed by five restriction endonucleases (Bam H I, Eco R I, Hind III, Sal I and Sma I) in comparison with prototype strain. Thirty-seven isolates were divided into six genome types (Ad37a, Ad37c, Ad37d, Ad37e, Ad37f and Ad37p). The predominant genome type in 1987 was Ad37a by DNA cleavage pattern analysis with Eco R I. The symptoms of patients with Ad37a were severer compared with those of patients with Ad37p.

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Since 1989, outbreaks of Salmonella ser. Enteritidis (S. Enteritidis) food poisoning have dramatically increased in Tokyo, and a total of 31 outbreaks has been reported in 1989. Twenty-one of these 31 outbreaks were caused by S. Enteritidis PT34, but 8 outbreaks were caused by S. Enteritidis PT4. After 1990 instead of SE PT34, food poisoning due to PT4, which was a very common phage type in the UK, has increased in Tokyo. Between 1989 and 1995, there were 144 food poisoning outbreaks caused by S. Enteritidis, and 64 of these outbreaks were by due to S. Enteritidis PT4, which was one of the main phage types in Tokyo.
To characterize these strains of phage type (PT) 4, 293 isolates from patients, and vehicle foods, eggs and environment in Tokyo were examined for plasmid DNA profiles, acid productivity from glycols (propylene and ethylene) and antimicrobial resistance patterns. Plasmid DNA was extracted by Kado's method, and analyzed by agarose gel electrophoresis. The acid productivity from plopylene glycol or ethylene glycol were tested using Barsicow medium with 1% propylene glycol or ethylene glycol. Antimicrobial susceptibility to AM, CP, TC, SM, KM, NA, ST, FOM and NFLX was tested by the K-B disc method.
The strains of PT4 were firter subdivided into 9 types by those epidemiologic marker analysis. The prevalent pattern of PT4 strains was type A plasmid profile carrying only one plasmid (60 kb) and there were 2 kinds of antibiograms. One was SM resistant, while the other was susceptible. A total of 56 (87.5%) of 64 outbreaks was found to have been caused by these types of S. Enteritidis.
Several kinds of egg-related foods were suspected as the vehicles of transmission among 24 outbreaks. Especially, in 5 outbreaks, S. Enteritidis strains were isolated both from patients and suspected food which were cooked with egg. This strongly suggests that these foods may be the potential source of infection in S. Enteritidis PT4 outbreaks.

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Anti-bacterial activities of a sulfonated human immunoglobulin preparation against penicillin resistant Streptococcus pneumoniae and Pseudomonas aeruginosa were examined. Five week old CBA/J mice were challenged by 10 times of 50% lethal doses of penicillin sensitive (SP1) and resistant (SP2) strains of Streptopcoccus pneuminiae serotype 19, and were treated with a sulfonated human immunoglobulin preparation (hIg). Fifty % protective dose (ED₅₀) were 2-4mg hIg/ mouse. These doses were parallel with the challenged doses despite of the challenged bacteria, and it was calculated that 6×10⁶ bacteria were killed by one mg of hIg in both bacteria.
Minimal inhibitory concentrations (MIC) of piperacillin, flomoxef, imipenem, amikacin and ofloxacin against SP1 and SP2 were estimated under the presence of various concentrations of hIg. It was found that under the presence of over 10-20 mg/ml of hIg, SP1 and SP2 were not grown even without any antobiotic. That is, MIC of hIg itself against penicillin sensitive and resistant Streptococcus pneumoniae(HR) serotype 19 were 10-20 mg/ml. Since about 10⁶ bacteria/ml were used for this test, it was calculated that 5-10×10⁴ bacteria were killed by one mg of hIgin vitro. This result suggested that in vivo anti-bacterial activities of hIg could be 100 times higher than that in vitro. Synergistic or at least additive effects between hIg and all antibiotics tested were seen by the MIC. If hIg was 100 times effective in in vivo, these results suggested that hIg could improve the effect of chemotherapy in clinical cases.
Similar in vitro test were carried out for Pseudomonas aeruginosa, and synergistic or at least additive effects between hIg and all antibotics tested were also confirmed in Pseudomonas aeruginosa. This result suggested that hIg could be effective for clinical pseudomonas infection.

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An arbitrarily primed polymerase chain reaction (AP-PCR) DNA profile was applied to epidemiological analysis of Salmonella enterica subsp. enterica serovar Enteritidis.
A total 21 strains of S. Enteritidis isolated from 21 cases (10 cases of healthy persons, 7 cases of food poisoning outbreaks and 4 sporadic diarrhea cases), during the period between December 1991 and August 1996 in Wakayama City, were used. A total of 60 arbitrary primers (DNA oligomer (12) set, Wako) were screened with 4 S. Enteritidis strains of different cases. A-11, B-32, C-42 and C-45 primers were chosen. Plasmid DNA profiles, antimicrobial susceptibility patterns and phage types were also examined.
The combination of these three methods resolved the collection into five groups (A to E). And type C strains were found in 17 cases (81%) out of 21 cases. However, according to AP-PCR DNA profile, all 21 strains were classified into six groups (I to VI), and 17 type C strains were classified into three groups (III, IV and V). Type IV was predominant in Wakayama City, and type C·EIV was found in 15 cases (71%).
In conclusion, we considered that AP-PCR DNA profile using appropriate primers was an effective epidemiological marker.

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In recent years there have been a number of subjects who were bitten by supposed rabid animals in foreign rabies-epizootic countries and visited our hospital to receive post-exposure therapy after their return to Japan. WHO recommends immediate washing of the wound with soap and water, applicatun of human anti-rabies immunoglobulin and administration of tissueculture rabies vaccine at 0, 3, 7, 14, 30, and 90 days after exposure. However, tissue-culture vaccines, are expensive and they are not always used in all parts of the world. The author checked whether the victims of animal bite were injected with rabies vaccines abroad or not and investigated the type of rabies vaccine when they were vaccinated. About a half of the consulted victims were locally injected with rabies vaccine. By mean of certificates of inoculation or empty boxes of vaccine, types of rabies vaccines were proved in 40 subjects of which 38 received tissue-culture vaccines. Semple-type vaccine was administered to one subject and suckling mouse vaccine was done to another one. When post-exposure prophylaxis was continued after return to Japan, it is important to know the sort of rabies vaccine injected abroad, because brain-tissue vaccines are less effective in inducing antibody than tissue-culture vaccines. Consequently both physicians and travelers should keep in mind that even now brain-tissue vaccines are used in some areas of the world.

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To study antigenic differences in Helicobacter pylori (H. pylori), we performed immunoblot (IB) analysis with monoclonal antibody, MAb102, and measured the serum antibody titer with captured ELISA method with MAb102 antibody. In IB analysis, specific 54 K antigen was detected in 39 strains derived from patients with gastritis, gastric ulcer, duodenal ulcer, and gastric carcinoma. But only one strain (GC32 strain) isolated from a patient with gastric cancer did not react with MAb102 antibody. In the result of antibody titer with captured ELISA method using other six strains of H. pylori as antigen, H. pylori strains were divided into 3 groups, group R (RD26, T7, and T13 strains), group E (England and MR31 strains), group T (T37 strain). However, four cases belonged to false negative in ELISA method. These cases could have been infected with H. Pylori strains such as GC32 strain. These strains did not have 54 K antigen nor 54 K related antigen which is different in its antigenicity from the 54 K antigen of the six strains. Thus the GC35 strain and the strain (s), which infected these four cases, consisted of other group, group N. These results suggested that 54 K antigen is usually stable in many strains, but occasionally different in antigenicity in some strains.

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Contamination of 24 hr home bath with Legionella pneumophila is recently well recognized. Eradication of the water-bath contamination from L. pneumophila and other bacteria is an important matter to prevent the infection because the 24 hr-bathing facility is widely accepted in Japanese houses. Among the 16 bathing water samples we tested, Legionella pneumophia was isolated from 6 cases (37.5%) when the bething water was not treated with disinfectants. Number of L. pneumophila increased up to 10³ cfu/ml and total culturable bacterial counts reached to 10⁵ cfu/ml within 5 days when the water was not treated. We selected 5 water bathes among 6positive cases to study the bactericidal effect of chorine. As a result we concluded that the growth of L. pneumophila in 24 hr-water bath could be stopped by the 2 ppm chlorination program every day.

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Efficacy and safety of a newer injectable cephalosporin, cefluprenam (CFLP) on cases with bacterial pneumonia and chronic respiratory tract infections were evaluated at a dose of 1g (potency), d.i.d for 7 days.
1. Of 130 cases in total, 116 cases were enrolled for the clinical efficacy evaluation. The efficacy rate (excellent and good responses) was 94.8%(110/116). The efficacy rate was 93.8%(60/64) for cases with bacterial pneumonia, and 96.2%(50/52) for cases with chronic respiratorytract infections.
The recurrence was noted in 1.2%(1/82).
The bacteriological response rate was 100.0%(32/32) for gram possitive cocci, 93.8%(15/16) for gram negative rods and 97.9%(47/48) in total.
2. Adverse drug reactions were noted in 3.9%(5/129), consisting of 2 cases with skin rash, 1 case with drug fever, 1 case with skin rash and skin itching and 1 case with drug fever and headache.
The abnormal laboratory changes were noted in 23.6%(30/127), mainly containing the elevation in GPT and GOT, and eosinophylia.
The safety rate (no problem evaluation) was 74.8%(95/127).
3. The usefulness rate (very useful and useful evaluations) was 93.1%(108/116).
As suggested by the evaluation on the secondary endpoint in the phase III comparative studies with both bacterial pneumonia and chronic respiratory tract infections, it was confirmed that the 7 day therapy of CFLP was promising for treatment of moderate bacterial pneumonia and chronic respiratory tract infectons, because the high clinical efficacy was obtained and also the incidence of allergic reactions with CFLP was almost the same as that of ceftazidime (CAZ) evaluated highly safe. Based on these results, it was concluded that CFLP was useful in the management of moderate respiratory tract infections and also the recommended therapeutic period with CFLP was within 7 days.

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A 25-year-old male admitted to Kawasaki municipal hospital with the diagnosis of typhoid fever. He had noticed high fever since one month ago, and had been treated with prednisolone with the diagnosis of fever of unknown origin in a hospital. Then he had admitted to St. Marianna University Hospital, and Salmonella Typhi had been detected from his blood and stool.
On admission, multiple liver abscess were detected by abdominal ultrasonography. S. Typhi in bile was not eliminated with CP and AMPC, but he was successfully treated with cholecystectomy and the chemotherapy of LVFX. Abscess formation was found in the resected gall bladder wall.
Typhoid nodule in the lymphnode, liver or other organs is a well known pathological change in the typhoid fever. But abscess formation in the liver or other organs is rare. In this case, multiple abscess is characteristic and this cause is thought to be induced by the factors that the period from onset of typhoid fever to beginning of effective therapy was too long, and that steroid therapy was done without antibiotic therapy.

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