Cholera toxin gene and production of enteroreactive toxins were examined in 134 strains of non-O1 V. chrolerae. Results obtained were summarized as follows. Frequencies of cholera-toxin-genepositive strains were 2/58 (3.4%) from human sources and 2/76 (2.6%) from fish and environment. While, frequencies of production of hemolysin, fluid accumulating factor (FAF) related with protease, fluid accumulating factor in the suckling mouse, NAG-rTDH, NAG-ST and Vero toxin were 100, 72, 31, 2, 0 and 0%, respectively, for 58 strains from human sources, and 100, 57, 24, 0, 1.3 and 0%, respectively, for 76 strains from fish and environment. Among the 31 strains used for the injection of viable cells to the ligated rabbit ileal loop, detection frequencies of these enteroreactive toxins in the accumulated fluids were 100% for hemolysin, 3.2% for both FAF and NAG-rTDH and 0% for cholera toxin, Vero toxin or NAG-ST. Hemolysin and the fluid accumulating factor in the suckling mouse seemed to be identical in most strains. These results suggest that cholera toxin, NAG-ST, NAG-rTDH and Vero toxin may not be very important in the enteropathogenic mechanism of a great majority of non-O1 V. cholerae strains, whereas hemolysin may play an important role in the enteropathogenicity.
An outbreak of cholera occurred among guests of the NTT Nagoya Hall in September 1989. Clinical findings showed that all but one were symptomatic infections out of 44 bacteriologically confirmed cases. In relation to the high incidence of symptomatic infections, we examined cholera toxin (CT) producibility of the isolated V. cholerae. 1. Strains of the NTT case produced 16-256 (mean 130) ng of CT per ml in CAYE-L medium at 30°C and 32-256 (mean 142) ng of CT per ml by Polymyxin B treatment. But strains of past case produced 8-256 (mean 34), 8-128 (mean 44) ng of CT per ml, respectivity. Strains of the NTT case produced a larger amount of CT than that of the past cases. 2. Strains of the NTT case produced 512-4096 (mean 2100) ng of CT per ml in CAYE-L medium at 37°C and 1024-2048 (mean 1600) ng of CT per ml by Polymyxin B treatment. But strains of the past case produced 8-64 (mean 25), 8-128 (mean 45) ng of CT per ml, respectivity. Strains of the NTT case produced a larger an amount of CT than them of past case. We observed the same phenomenon in AKI medium at 37°C. The yield of CT in CAYE-L medium was greater at 37°C than 30°C. 3. Strains of NTT case grew faster than that of the past case in CAYE-L medium at 37°C. But the growth rate was the same as both strains in AKI and CAYE media. 4. V. cholerae grew in the presence of Lincomycin produced elevated levels of CT.
Eleven patients with acute non-lymphocytic leukemia developed hepatosplenic candidiasis following the termination of chemotherapy. Five who did not receive amphoteicin B (AMPH) died of liver failure, whereas the other six who received AMPH and/or miconazole or fluconazole were cured.The features of hepatosplenic candidiasis included prolonged high fever not responsive to antibiotics and hepatomegaly with right-sided hypochondrial pain developing after severe neutropenia. Even if the neutropenia recovered, these symptoms did not subside. In addition, elevation of the serum alkaline phosphatase and total bilirubin levels were observed at onset. CT scan revealed multiple low-density areas of varying size that showed no contrast enhancement. Ultrasonography also demonstrated hypoechoic or hyperechoic lesions, and a so-called “target sign appearance” in the liver and spleen. The clinical diagnosis for hepatosplenic candidiasis is not so difficult because of the uniform symptoms, signs and laboratory abnormalities. The importance of the early administration of antifungal agents to obtain a cure is discussed in this article.
Human herpesviruses have been known to be reactivated from latent infection in immunosuppressed individuals. To asses the immunity, antibodies to human herpesviruses (VZV, HSV, CMV and EBV) were measured in patients with gastointestinal cancer and normal persons ranging from 40 to 80in age. CF antibodies to herpesviruses except for EBV were negative in normal persons below the age of 60 and cancer patients aged 60 and over. The CF antibody titer higher than 1: 64 occurred only in young cancer patients and older normal persons. The titer of VCA antibody to EBV ranged from 1: 40 to 1: 160 in more than 80% of normal persons and was higher than 1: 640 in less then 7%. The ratio those with antibody negative or below 1: 40 increased with the age. In contrast, the titer was higher than 1: 320 in 70% of the cancer patients, and the antibody was negative in none of them. In addition, the low titer occurred only in less than 5%. The rate of patients having higher VCA antibody titers increased with age, and the titer was high in more than 50% of patients in the 8th decade. In cancer patients, the higher the VCA antibody titer was, the less number of mature B cells, target cells of EBV, and the B cell functions were observed. This suggests that reactivation of EBV inhosts bearing carcinomas results in a fall of production of the specific antibody.
We evaluated the efficacy of Itraconazole (ITZ), a newly developed triazole antifungal agent, in 5 cases of pulmonary aspergilloma. A daily dose of 100-200 mg of ITZ was given orally for 5 to 20 months. In one patient, administration of ITZ was discontinued because of mild and transient hepatic injury. On computerized tomography and conventional roentgenogram of the chest, improvement was clearly observed in 2 of the remaining 4 cases. Symptomatic improvement, mainly disappearance of airway bleeding, was seen in 2 cases. Thus the overall improvement rate was 50%(two out of 4). No adverse response other than the hepatic injury mentioned above occurred. In one patient, who had two fungus balls previously, one of the fungus balls disappeared completely and the other decreased significantly in size during the course of treatment. In this case, the daily dose of ITZ was larger (200 mg once a day) and the plasma concentration of ITZ was extremely higher (2068ng/ml) than in other cases (137-330ng/ml). We conclude that ITZ is an efficacious drug in the treatment of pulmonary aspergilloma, and that this drug may be even more efficacious if we keep higher plasma concentration of ITZ.
We succeeded in isolating Aeromonas-susceptible phages from river water and mud after a few years (1983-1985) of painstaking effort. For the first time in Japan, we investigated phagetypes of the bacterial strains isolated. The results obtained are summarized as follows: 1) Aeromonas-susceptible phages were isolated from 82 (40.1%) of 195 samples of river water and from 23 (25.6%) of 90 samples of river mud. By the cross-matching test of the phages isolated from the 105 samples, these were classified into 13 type groups (Groups I-XIII) 2) When 594 Aeromonas strains isolated from river water, lake water, river mud and fresh-water fish were examined using the phage group patterns developed by us, 129 (21.7%) strains classified into these phagetypes. Phagetyping was possible for 11 (15.5%) of the 71 strains isolated from river and lake water, for 29 (35.4%) of the 82 strains from river mud and for 89 (20.2%) of the 441 strains from fresh-water fish. By bacterial species, phagetyping was possible for 53 (51.5%) of the 103 strains of A. hydrophila. 21 (7.2%) of the 292 strains of A. sobria, 13 (8.8%) of the 148 strains of A. caviae and 42 (82.4%) of the 51 strains of Aeromonas spp. Especially the phages classified as Groups I, IV and VI amounted to the majority. Thus we succeeded in isolating Aeromonas-susceptible phages which could be classified into Groups I-XIII. The results suggested the possibility of utilizing this phagetyping for analysis of the ecological distribution of genus Aeromonas.
Among the group A streptococci isolated during the period 1956 to 1978 from the pharynx and the nostril of the scarlet fever patients quarantined in the Tokyo Metropolitan Toshima Hospital within 1 week of hospitalization, 1, 586 strains of types 4, 6 and 12, the most prevalent serotypes, were estimated for their sensitivities to various antibiotics. Among the sensitivities, those to tetracycline (TC), chloramphenicol (CP) and erythromycin (EM), these being in close relation to prevalent serotypes, are to be reported. The first appearance of TC-resistant strain was seen in one strain (type 6) isolated in 1959, and, in 1964, the resistant strain increased in proportion in accordance with the prevalence of type 4. The resistant strain decreased a little in 1968 and in 1969, increased again from 1970, exceeded over 90% from 1973, and reached 100% in 1978. The first appearance of CP-resistant strain was seen in one strain (type 4) isolated in 1969, and then the resistant strain increased year by year along with the prevalence of type 12. The first appearance of EM-resistant strain was seen in 30 strains (type 12) isolated in 1972, and then, similarly to the case of CP-resistance, the resistant strain increased year by year together with the prevalence of type 12. Meanwhile, among the 1, 586 strains isolated at the time of hospitalization and estimated of their minimum inhibitory concentrations (MIC), 932 strains (58.8%), 452 strains (28.5%), and 574 strains (36.2%) showed resistance to TC, CP, and EM, respectively. Variations of drug-resistance by serotypes were as follows: TC-resistant strains increased in accordance with the prevalence of type 4 in 1964; CP-and EM-resistant strains increased in accordance with the prevalence of type 12 in 1972; and type 6 strains were low in the rate of resistant strain to any of the three drugs. So far, prevalence of a serotype of group A streptococci has been considered to be caused by the accumulation of the subjects non-immune to the serotype. In the present study, a phenomenon was observed that prevalence of a serotype was effected by the state of resistance-acquisition to commonly used drugs. It would be clear that the resistance-acquisition plays, in addition to the immune state, a great role in evoking prevalence of a serotype.
Enterotoxigenic strains of Vibrio cholerae O-1 biotype eltor, isolated from three sporadic cases of cholera in Nagoya in 1989 and an outbreak of cholera in Nagoya in 1989 were analyzed for their similarities. All isolates of V. cholerae O-1 were indistinguishable in bacteriophage types, serovars, biovars and drug resistance patterns. Because the epidemiological investigation based on a primary structure of chromosomal DNA is more reliable, we isolated chromosomal DNA from these isolates and compared electrophoretic patterns of restriction endonuclease-digested DNA fragments. Furthermore, Southern hybridization of the cholera toxin gene was performed. Since no difference among six strains in these sporadic was observed, it was strongly suggested that both the independent cases and the outbreak of cholera were caused by the same strain.
We compared the lysis centrifugation blood culture system using IsolatorR (Du Pont) to the conventional blood culture bottle using BCB system ® (Roch), and the following results were obtained. In the investigation of experimental fungemia made by human blood and Candida albicans, the organism was detected 9/12 after 1 day and 11/12 after 3 days by Isolator, although the organism was detected 3/12 after 3 days and 11/12 after 6 days by the culture bottle system. In the experimental fungemia of Candida tropicalis, the organism was detected 12/12 after 2 days by Isolator, although it was detected 1/12 after 6 days and 4/12 after 10 days with the culture bottle system. Isolator showed faster and a more sensitive detection rate for fungemia. Clinical investigation of 94 cases (180 cultures) showed a 11.1% isolation rate with Isolator system and a 7.1% isolation rate with the culture bottle system. In 43 cases which were examined by the both systems, 7 cases were positive with Isolator, 3 positive with culture bottle. The culture bottle system isolated only Candida albicans and Candida parapsilosis, where as the Isolator isolated Candida albicans, Candida parapsilosis, Candida tropicalis, Candida krusei, and Candida spp. This investigation suggested that the Isolator system was an excellent method for the detection of fungemia.
The present study was undertaken to establish a novel detection method for Neisseria gonorrhoeae (N. gonorrhoeae). DNA probe obtained from Gen-probe Inc. was employed and clinically isolated strains (18 species, 81 strains) were applied to verify the quality and quantity of the method. The present data revealed that the method is specific to detect N. gonorrhoeae (30/30): neither gram positive nor negative oraganisms were detectable by the method (0/51). It has been shown, further more, that there exists a significant correlation between the number of the bacteria and the relative light units (r=0.96). The threshold of cell numbers, on the other hand, was found to be 5 × 103 cfu/tube. The detection sensitivity for N. gonorrhoeae was not influenced by the presence of other bacteria, and not decreased by using heat-treated or drug-treated N. gonorrhoeae organisms. The present study also provides some experimental evidences that the method can also be applicable for the detection of bacteriolytic action of drugs.
We determined the incidence of herpes zoster (HZ) in 119 patients with systemic lupus erythematosus (SLE). HZ occurred in 56 patients (47%), and 9 patients had had HZ even before SLE developed. After diagnosis of SLE, an incidence of zoster was high, 5.45 cases per 100 person-years. It was found that the susceptibility to HZ was not related to the presence of renal disorder or maximum dose of corticosteroids. The patients with SLE who had had HZ showed significantly higher antibody titers than those without a history of HZ and normal subjects as assayed by both complement fixation technique and neutralization test. On the other had, only 17 of 55 patients (31%) with SLE showed positive skin reactions to varicella zoster virus (VZV) antigen, whereas all 15 normal subjects had positive reactions. In the patients who were receiving less than 10 mg/day of prednisolone, 11 of 17 (65%) had positive skin reactions to VZV antigen, whereas only 4 of 31 (13%) patients who were receiving 10 mg/day or more prednisolone showed positive reactions. It was of interest that in 7 patients with SLE who had not received corticosteroids, only 2 (29%) patients showed positive skin reactions to VZV antigen. These results suggest that high incidence of HZ in patients with SLE is probably due to an impaired cellular immunity because of both underlying disease and corticosteroid treatment.
A total of 1, 189 Shigella strains consisting 804 imported and 385 domestic strains were examined for their species and serovar distribution, and their drug-resistance. In both imported and domestic strains, S. sonnei was found to be the most prevalent species, followed by S. flexneri, S. boydii and S. dysenteriae in order. In imported strains, however, the isolation frequency of S. flexneri, S. boydii and S. dysenteriae were higher than that of domestic strains, and the serovar of each species was distributed in a wider range than that of the domestic strains. The drug resistance test using chloramphenicol (CP), tetracycline (TC), streptomycin (SM), kanamycin (KM), ampicillin (ABPC), sulfamethoxazole-trimethoprim (ST), nalidixic acid (NA), fosfomycin (FOM) and norfloxacin (NFLX) showed that 80.1% of the imported strains and 82.9% of the domestic strains were resistant to any of drugs examined, although frequency of resistance varied by their species. The frequency of resistance to each drugs were SM (74.0%), TC (68.5%), CP (38.8%), ABPC (35.3%), ST (34.1%), NA (1.5%) and KM (1.0%) in imported strains, and TC (79.2%), SM (55.3%), ABPC (40.3%), CP (38.4%), ST (32.4%), NA (30.5%) and KM (4.2%) in domestic strains. No strain resistant to FOM or NFLX was found in both groups. The resistance patterns varied to 32 types. Among those, a multiple drug-resistance type with CP·TC·SM·ABPC was found most frequently in both groups. Frequency of strains carrying transferable R plasmid was tested for 50 strains isolated in 1989 and three (6%) of them had the plasmid.
Enteropathogenic micrroorganisms isolated from feces of 9, 393 patients with diarrhea or enteritis in our hospital between 1976 and 1988 were analyzed As the result of the examination of 5, 443 outpatients, 1, 811 strains of pathogens were isolated from 1, 686 cases (31.0%). Several species including Salmonella spp., Escherichia coliserotype, Vibrio parahaemolyticus, were isolated before 1978, and the incidence of pathogens was low (14.8%). For the 10-year period since 1979, the incidence markedly increased to 34.4%, and the number of pathogens isolated also increased to about twice that before 1978. The main cause of the increase wasCampylobacterspecies. The major pathogens detected since 1979 wereCampylobacterspp., E. coliserotype, Salmonellaspp., V. parahaemolyticus, etc., but Rota virus, Clostridium difficile, Aeromonasspp., Vibrio fluvialis, etc. have also been detected, showing an increase in the number and diversity of the detected pathogens. As the result of the examination of 3, 950 inpatients, 835 strains of pathogens were isolated from 800 cases (20.3%). The incidence ofC. difficilewas the highest, 423 of 800, followed byE. coliserotype, Salmonellaspp., Campylobacterspp., V. parahaemolyticusandAeromonasspp., in that order. All the inpatients from whomC. difficilewas isolated manifested diarrhea or enteritis afteradministration of antimicrobial agents. The pathogens causing communicable disease wereSalmonellaspp. serovar Typhi, Salmonellaspp. serovar Paratyphi A, Shigella flexneri, Shigella sonneiandEntamoeba histolytica, which were isolated from 5, 1, 3, 2 and inpatients, respectively.
Slime production was studied in a total of 598 strains of Staphylococcus aureus and 11 species of coagulase negative staphylococci and the relationship of the slime production and biofilm formation was investigated The high slime production was obtained in 42 of 263 strains (16%) of S. epidermidis only. The adherence ofS. epidermidisto biomedical silicone discs (S discs) and biofilm formation were investigatedin vitroin high slime producing and non-producing strains. The adherence rates ofS. epidermidisto the S discs ranged from 2.3 to 14.8%. There was no relationship between slime production byS. epidermidisand the adherence rates. Biofilm was formed only in slime producing strains with cell growth on the S discs when they were incubated. The process of the biofilm formation was obtained by scanning electron microscopy; there were many projections, probably slime secreted from the cells, on the cell surface, and picture showing agglutination of cell by crosslinking. The bactericidal activity of cefazolin, cefmetazole and flumoxef againstS. epidermidisafter formation of the biofilm decreased more markedly than that before formation. Cefazolin reduced the number ofS. epidermidisand eliminated the formed biofilm in anin vitrosimulation model of serum concentration in human after drip infusion of 2 g of cefazolin.
A 88 year old Japanese male was repeatedly infected withAncylostoma duodenale.He underwent an artificial anus operation about 55 years ago. He appeared to be infected with hookworm earlier than in 1977 and developed severe anemia. Though he was treated with pyrantel pamoate and mebendazole several times, reinfections developed in each time. A possible origin for his reinfections was his own feces defecated through his artifical anus. Unsanitary handling of the anus and the feces exposed himself to oral or percutaneous infection. Besides, a single dose of pyrantel pamoate, usually very effective against Ancylostoma duodenale, was not so effective in this patient. Therefore, we prescribed multiple doses of pyrantel pamoate, and followed by a single dose of mebendazole. However, reinfections still persisted because of his unsanitary behavior.
Increasing rates of human immunodeficiency virus (HIV) related tuberculosis have been noted and recently the clinical importance of the disease has been mentioned. The diagnosis of tuberculosis is more difficult in those patients with HIV seropositive than those with seronegative, because those with seropositive have atypical clinical features A 29-year-old male, who was infected with HIV heterosexally in Central Africa in 1986, was admitted to our hospital with a history of general malaise and weight loss in April, 1989. Laboratory and physical examinations revealed anemia, thrombocytepenia, the elevation of LDH, and giant intraabdominal lymphadenopathies, suspecting malignant lymphoma.Mycobacteriumwas isolated from the sputa in April and was confirmed asM. tuberculosisusing a DNA probe in May, 1989. Clinical symptoms including giant lymphoadenopathies and laboratory abnomalities improved with antituberculosis therapy. Development of a rapid method for the diagnosis of tuberculosis was warranted in this case.