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IS231-MIC231 elements from Bacillus cereus sensu lato are modular.

Summary IS231A was originally discovered in Bacillus thuringiensis as a typical 1.6 kb insertion sequence (IS) displaying 20 bp inverted repeats (IR) flanking a transposase gene. A first major variation of this canonical organization was found in MIC231A1. This mobile insertion cassette (MIC), delineated by IS231A-related extremities, contained an active d-stereospecific endopeptidase (adp) gene instead of a transposase. Interestingly, it was shown that MIC231A1 can be mobilized in trans by the IS231A transposase. In this paper, we show that this family of IS231-MIC231 elements can be extended to a broad range of related entities displaying higher levels of structural complexity. Several IS231A-like elements contained, upstream of their transposase gene, passenger genes coding for putative antibiotic resistances or regulatory factors. Furthermore, the diversity of the MIC231 elements ranged from empty cassettes to structures carrying up to three passenger genes. Among these, MIC231V carried, in addition to an adp gene, an active fosfomycin resistance determinant. In vivo transposition assays showed that MIC231V is also trans-activated by the IS231A transposase. These results lend further support to the potential contribution of these modular mobile elements to the genome plasticity of the Bacillus cereus/B. thuringiensis group.

Bacillus cereus↗

Identification of Bacillus subtilis sigma-dependent genes that provide intrinsic resistance to antimicrobial compounds produced by Bacilli.

Bacillus subtilis produces many antibiotics of varying structures and specificity. Here we identify a prominent role for sigma(W), an extracytoplasmic function (ECF) sigma factor, in providing intrinsic resistance to antimicrobial compounds produced by other Bacilli. By using a panel of B. subtilis mutants disrupted for each of the 30 known sigma(W)-dependent operons we identified resistance genes for at least three different antimicrobial compounds. The ydbST and fosB genes contribute to resistance to antimicrobial compound(s) produced by B. amyloliquefaciens FZB42, the yqeZyqfAB operon provides resistance to the SPbeta prophage-encoded bacteriocin sublancin, and the yknWXYZ operon and yfhL provide resistance to the antimicrobial peptide SdpC. YfhL encodes a paralogue of SdpI, a membrane protein that provides immunity to SdpC. In competition experiments, we identify sigma(W) as a key factor in allowing B. subtilis to resist antibiotic killing and encroachment by competing strains. Together with the previous observation that sigma(W) provides inducible resistance against the Streptomyces antibiotic fosfomycin, these studies support the notion that sigma(W) controls an antibiosis regulon important in the microbial ecology of soil bacteria.

Amino Acid Sequence↗

Bacterial meningitis caused by Veillonella parvula.

A three-year-old girl injured her right eyelid with a toothbrush. The wound was sutured. Swelling of the eyelid, high fever and vomiting developed in spite of oral antibiotics for seven days. The findings of the cerebrospinal fluid (CSF) were white blood cells (WBC) 26,368/mm3 (90% polymorphs), protein 127 mg/dl, and sugar 0 mg/dl. Although Gram negative organisms were seen on the smear, aerobic culture was sterile. Later culture of CSF on admission grew anaerobic bacteria: Veillonella parvula. Intravenous administration of penicillins with cefotaxime (CTX), or of fosfomycin (FOM) were ineffective. Chloramphenicol (CP) cured the patient without neurological sequelae. There were no abnormal findings on brain CT scan. This is the first report of Veillonella meningitis. V. parvula appeared to have invaded the CSF from the abscess of the eyelid. It is necessary to consider anaerobic meningitis when there is a preceding pyogenic infection in the head.

Bacterial Infections↗

Methicillin-resistant Staphylococcus aureus (MRSA) with reduced susceptibility to glycopeptides (GISA) in 63 French general hospitals.

Clinical isolates of methicillin-resistant Staphylococcus aureus (n = 1070) collected from 63 French general hospitals during June 2000 (n = 1070) were screened initially for reduced susceptibility to glycopeptides (GISA) on brain-heart infusion agar containing teicoplanin 6 mg/L. Glycopeptide MICs were determined for the 145 isolates that grew on the screening plates. Of the 1070 isolates, 1.4% were GISA on Mueller-Hinton agar, and 2.9% by Etest with a high inoculum, while 0.7% and 2.9% were GISA by vancomycin and teicoplanin population analysis profiles, respectively. Most isolates were resistant to gentamicin and rifampicin or fosfomycin or fusidic acid, as determined by disk diffusion. Pulsed-field gel electrophoresis of the 31 GISA isolates identified four clones, with dissemination of one predominant clone. In these French hospitals there was a low incidence of GISA and hetero-GISA.

Anti-Bacterial Agents↗

Evaluation of the OSIRIS video reader as an automated measurement system for the agar disk diffusion technique.

Measurement of inhibition zones by the automated OSIRIS system was compared with manual measurement. In total, 14 176 measurements were made with 352 staphylococcal and 80 Enterobacteriaceae isolates, involving four panels of antibiotics on round and square Mueller-Hinton agar plates, according to the German DIN 58940 recommendations. Variations of +/- 3 mm in zone size measurements were defined as tolerable. Very major errors (i.e., classification of a resistant isolate as susceptible by the OSIRIS system) occurred in < 1% of tests. With staphylococci, the best concordance was recorded for rifampicin (91.3%), moxifloxacin (88.1%), and gentamicin (86.3%), while the concordance on square plates for vancomycin, pristinamycin and kanamycin was 97.2%, 96.1% and 96.0%, respectively. The poorest concordance was for cefuroxime (43.7%) and novobiocin (47.0%) on round plates, and fosfomycin (36.5%) and chloramphenicol (84.0%) on square plates. With Enterobacteriaceae, 100% concordance was recorded for ampicillin, gentamicin and ciprofloxacin on round agar plates, and for gentamicin, cefoxitin and nalidixic acid on square plates. The poorest results were recorded for nalidixic acid (32.5%) and piperacillin (82.5%) on round plates, and for nitrofurantoin (72.5%) and amoxycillin (82.5%) on square plates. It was concluded that the OSIRIS system was a rapid and reliable system for measuring disk susceptibility test results on round and square agar plates.

Anti-Bacterial Agents↗

Drug resistance and random amplified polymorphic DNA analysis of Photobacterium damselae ssp. piscicida isolates from cultured Seriola (yellowtail, amberjack and kingfish) in Japan.

AIMS: To investigate the antimicrobial susceptibility and genetic characteristics of Photobacterium damselae ssp. piscicida isolates obtained from cultured Seriola in Japan. METHODS AND RESULTS: Minimal inhibitory concentrations of 14 antimicrobials for 74 isolates from Seriola in Japan in 2002 were determined. Isolates showed high frequencies of resistance to sulfamonomethoxine (SMMX) (97.3%), oxytetracycline (OTC) (77.0%), flumequine (FMQ) (77.0%), chloramphenicol (CP) (75.7%), kanamycin (KM) (63.5%) and oxolinic acid (OA) (62.0%), but low to ampicillin (ABPC) (2.8%). All isolates were susceptible to bicozamycin (BCM), fosfomycin (FOM) and florfenicol (FF). Of these isolates, 45 (60.8%) showed same resistance pattern (SMMX-OTC-FMQ-OA-CP-KM). In random amplified polymorphic DNA (RAPD) analysis, no difference was observed among our 74 field isolates and ATCC51736 isolated from Seriola in 1974 in Japan, but different from ATCC 17911 isolated from white perch in USA. CONCLUSIONS: FF, BCM, FOM and ABPC were useful antimicrobials for treating pseudotuberculosis. However, the frequency of multidrug resistance was high. RAPD analysis showed homogeneity of isolates from Seriola in Japan. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that some antimicrobials were still useful for treating pseudotuberculosis and that P. damselae ssp. piscicida strains of same origin might have spread among Seriola in Japan since 1974.

Animals↗

Antimicrobial resistance of Listeria spp. recovered from processed bison.

AIMS: The current study examined the antimicrobial susceptibility of 86 Listeria spp. isolated from processed bison carcasses. MATERIALS AND METHODS: Susceptibility to 25 antimicrobial agents was determined using E-test and National Antimicrobial Resistance Monitoring System (NARMS) panels. Most Listeria isolates (88-98%) exhibited resistance to bacitracin, oxacillin, cefotaxime, and fosfomycin. Resistance to tetracycline (18.6%) was also common. Of the 16 tetracycline-resistant Listeria isolates, 15 carried tetM and 2 contained integrase of Tn1545 transposons. Rifampicin and trimethoprim-sulfamethoxazole were the most active antimicrobial agents against Listeria spp., with a MIC(90) of 0.38 microg ml(-1). Ampicillin, erythromycin, penicillin, gentamicin, and tobramycin also exhibited good activity against Listeria spp., with MIC(90) not exceeding 1 microg ml(-1). Differences in resistance among Listeria spp. was displayed, as Listeria innocua strains were more resistant than other Listeria species. CONCLUSIONS: The study showed that Listeria monocytogenes strains from bison were susceptible to the antibiotics most commonly used to treat human listeriosis. However, the presence of antimicrobial resistance in L. innocua indicates the potential for transfer of resistance and a conjugative transposon to L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of our study will provide useful information for the development of public health policy in the use of antimicrobials in food animal production.

Animals↗

[Treatment of erysipelas in Germany and Austria--results of a survey in German and Austrian dermatological clinics].

BACKGROUND: Erysipelas is a severe soft tissue infection usually caused by streptococci. The infection is restricted to the dermis and subcutaneous tissues. Treatment with antibiotics is essential. Many different therapeutic regimens are recommended, based mainly on empirical data and only partly proven by clinical studies. MATERIAL AND METHODS: Our aim was to evaluate the treatment of erysipelas in Germany and Austria by means of a questionnaire and to derive treatment recommendations from this data. RESULTS AND CONCLUSION: The majority of clinics treat patients with erysipelas as inpatients with intravenous antibiotics. The usual first line treatment is group G penicillin (80%). Other choices include amino-penicillins (11%), cephalosporins (16.5%) and anti-staphylococcal penicillins (6.9%) are used. As second line antibiotics macrolides (63.5%), clindamycin (52.5%), penicillins (18.5%), cephalosporins (40%) and fluoroquinolones (20.5%) are mentioned. Carbapenems, tetracyclines, nitroimidazoles, glycopeptides, aminoglycosides, cotrimoxazole, fusidic acid and fosfomycin are used rarely. The median treatment duration is 10 days. Adjuvant measures are anticoagulation, non-steroidal anti-inflammatory agents, dressings, immobilization and treatment of local predisposing factors such as interdigital tinea.

Ambulatory Care Facilities↗

Susceptibility testing of Danish isolates of Capnocytophaga and CDC group DF-2 bacteria.

Twelve Capnocytophaga and seven DF-2 strains were tested for their susceptibility to 14 antimicrobial agents using an agar dilution and an agar diffusion method. Twenty-three other antibiotics were evaluated using the diffusion test only. All strains were fully susceptible to penicillin, ampicillin, cefuroxime, cefotaxime, erythromycin, clindamycin, chloramphenicol, doxycycline, rifamycin and ofloxacin using both methods. Clindamycin, rifamycin and cefotaxime were most active. Using agar dilution some strains were susceptible to gentamicin, but agar diffusion showed total resistance. One Capnocytophaga strain was susceptible and another moderately susceptible to metronidazole, other strains were resistant. The agar diffusion test showed that both Capnocytophaga and DF-2 were resistant to most other aminoglycosides, to fosfomycin, polymyxin and trimethoprim. All strains of both taxa were fully susceptible to piperacillin, cefoxitin, imipenem and fusidic acid and showed different susceptibilities to the other agents. Susceptibility testing by means of agar diffusion using an enriched chocolate agar and 5% CO2 atmosphere could be used to test Capnocytophaga and DF-2 strains and gives sufficient accuracy for routine use, when revised inhibition zone breakpoints are employed.

Animals↗

[A case report of meningitis and sepsis due to Enterococcus faecium complicated with strongyloidiasis].

A 80 year-old male was transferred to our department on 18th Aug. 1988, for high fever and clouding of the consciousness. He had been treated with steroid hormone (betamethasone 3.0 mg/day for 15 days) for his uveitis. Enterococcus faecium was isolated from both blood and spinal fluid, and then Strongyloides sterocoralis was revealed both in the sputum and stool. Anti-Human T-cell leukemia virus 1 (HTLV-1) antibody was also positive serologically. At first, beta-lactam antibiotics were used for the treatment of purulent meningitis and sepsis, but after performing sensitivity tests for E. faecium, the antibiotics were changed to rifampicin (RFP), fosfomycin (FOM) and ofloxacin (OFLX) for their excellent activity against the organism. After the clinical symptoms, subsided, thiabendazole was used for disseminated strongyloidiasis in daily doses of 2,500 mg for six days initially. The drug was used three times with two week intervals. Both bacterial and parasite infections subsided and no recurrence has been noticed until now. This is the first case of meningitis caused by E. facium complicated with strongyloidiasis.

Aged↗

[Drug-resistance and its transferability of Shigella strains isolated in 1986 in Japan].

Two hundred and fifty Shigella strains isolated in Japan were examined for their drug-resistances and their conjugal transferability. Drugs used and the isolation frequency of resistant strains were: sulfanilamide (SA) 96%, streptomycin (SM) 85%, tetracycline (TC) 82%, fosfomycin (FOM) 76%, chloramphenicol (CP) 51%, trimethoprim (TMP) 46%, ampicillin (ABPC) 45%, nalidixic acid (NA) 6%, kanamycin (KM) 5%, cephaloridine (CER) 4%, gentamicin (GM) 0%, amikacin (AMK) 0%, colistin (CL) 0%, norfloxacin (NFLX) 0%. All of the KM-resistances and none of the FOM-resistances were transferable. Each 20 to 30% of CP, TC, SM, SA or ABPC-resistant isolates transferred each drug-resistance. Two thirds of the isolates were brought to Japan by travellers mostly from South-East Asia. The isolation frequency of resistant strains was not different between domestic and foreign strains except that NA-resistance was not found in foreign strains. The frequency of isolates carrying transferable R plasmids were higher in domestic strains than foreign strains. Most of the isolated had multiple drug-resistance. Major resistance types were: (CP, TC, SM, SA, ABPC) 22%; (TC, SM, SA, TMP) 20%; (CP, TC, SM, SA, ABPC, TMP) 16%; (TC, SM, SA) 8%. Strains showing different resistance types were different among each other in their serum type, conjugal transferability, and derived country. Stepwise evolution of multiple resistance in one country was not demonstrated. The resistance types of Shigella isolates in Japan have been influenced by yearly change of imported strains.

Anti-Bacterial Agents↗

[A case of septicemia due to Capnocytophaga ochracea beginning post radiation therapy for lung cancer].

A 46-year-old man was admitted to our hospital on Mar. 16, 1988 with the chief complaint of productive cough. The chest roentgenogram and tomogram showed a tumorous shadow in the right upper lobe, accompanied with stenosis of the trachea and the right main bronchus. Bronchofiberscopic examination confirmed a nodular tumor protruding into the lower part of the trachea from the right lateral wall and nearly complete obstruction of the right main bronchus. Pathological specimen obtained by transbronchial biopsy revealed "low differentiated adenocarcinoma". It was highly suspected that the primary lung cancer had directly invaded the trachea and the right main bronchus. His symptoms and roentgenological findings remarkably improved after radiation therapy. He was discharged on May 12. On Sep. 14, he was admitted to our hospital again because of hoarseness, general fatigue and increasing dyspnea. The chest CT demonstrated severe stenosis of the trachea, which was treated with another radiation therapy. Although his symptoms diminished, he had a sudden onset of high fever on Oct. 15. Immediately a blood culture and transtracheal aspiration (TTA) were performed. Gram-negative bacilli were isolated from the blood culture four days later. The administration of fosfomycin and tobramycin was started. However he died because of massive hemoptysis on Oct. 23. Several days after the death, an isolated strain was identified as Capnocytophaga ochracea by the biochemical characteristics. Culture of sputum obtained by TTA was negative for Capnocytophaga ochracea. Septicemia due to Capnocytophaga spp. is very rare and only one case (due to Capnocytophaga sputigena) has been reported until now in Japan. Our patient is thought to be the first case of septicemia due to Capnocytophaga ochracea in Japan.

Adenocarcinoma↗

[Biochemical characteristics, growth on selective media, antimicrobial susceptibility, and diarrheagenic toxin production of enteroinvasive Escherichia coli].

A total of 70 strains of enteroinvasive Escherichia coli (EIEC) belonging to 8 different O serogroups including O28ac O29, O121, O124, O136, O143, O144, and O164, was studied for their biochemical characteristics, growth on selective isolation agar, antimicrobial susceptibility, and diarrheagenic toxin production. Among the biochemical characteristics examined, all EIEC strains gave negative lysine decarboxylation and all but one belonging to O124 serogroup, were non-motile, regardless of their O serogroups. The one motile O124 strain had a H30 antigen. Some close correlations were also observed between their O serogroups and biochemicals such as utilization of sodium acetate and mucate, ornithine decarboxylation, arginine dihydrolation, gas production from glucose, and lactose fermentation. Among the selective isolation agars, MacConkey and Deoxycholate-hydrogen sulfide-lactose (DHL), and modified Shigella-Salmonella (SS) agars supported growth of the EIEC strains well, whereas SS agar was remarkably inhibitory for strains of some O serogroups. Antimicrobial susceptibility was tested for nine drugs including chloramphenicol (CP), tetracycline (TC), streptomycin (SM), kanamycin (KM), ampicillin (ABPC), sulfamethoxazole-trimethoprim (ST), nalidixic acid (NA), fosfomycin (FOM) and norfloxacin (NFLX). Forty-one of the 70 strains (58.6%) were found to be resistant to the 6 drugs, such as CP, TC, SM, KM, ABPC or ST. None of the strains were resistant to NA, FOM or NFLX. Among the resistant strains recognized, the strains which showed the resistant patterns of CP.TC.SM.ABPC, CP.TC.SM, TC.SM.ST, TC.SM, and SM appeared to be prevalent. None of the strains gave positive reactions for the production of diarrheagenic toxins of heat-labile enterotoxin, heat-stable enterotoxin, and verocytotoxins 1 and 2.

Anti-Bacterial Agents↗

[Species and serovar distribution, and drug resistance of Shigella strains isolated from imported and domestic cases in 1980-1989 in Tokyo].

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.

Ampicillin Resistance↗

[In vitro and clinical studies of MRSA infections in compromised hosts].

From January to December 1991, 47 clinical cases in when methicillin-resistant Staphylococcus aureus (MRSA) strains were isolate were investigated at our internal medicine ward. The MRSA infection rate was 57.4% (27/47). The mortality due to MRSA bacteremia was 75.0% (9/12) and that due to MRSA pneumonia was 57.1% (4/7). We think that MRSA infections must be treated by multiple antibiotics. At out institution, most of the patients were given a combination therapy of imipenem + fosfomycin or imipenemt + mynocycline. Although in vitro the MICs of imipenem did not show excellent activity against MRSA strains, in vivo these combination therapy including imipenem showed excellent activity against MRSA infections. We think that this result was due to the additive effect of the two drug combination. We determined the MICs of single antibiotics against MRSA strains. Most of the MRSA strains were sensitive to minocycline and arbekacin. All MRSA strains were sensitive to vancomycin we think that vancomycin is a highly useful drug to combat MRSA infection.

Adult↗

[Microbiological and clinical studies of vancomycin resistant Leuconostoc spp. and Pediococcus spp. isolated from septicemia patients].

We described three septicemia cases in which blood cultures yielded gram-positive cocci identified as Leuconostoc spp. and Pediococcus spp. Patients were three male adults aged 63 to 71 years with severe underlying diseases, pancreatic cancer, esophageal cancer and diabetes mellitus with chronic renal failure. They had fever and chills at the onsets of septicemia with acute obstructive suppurative cholangitis, acute pneumonia, and infection complicated with invasion sites of esophageal cancer contagious to bronchus and subcutaneous tissue. Blood cultures yielded catalase and oxidase negative highly vancomycin-resistant (MIC: 1024 micrograms/ml <) gram-positive cocci showing alpha or gamma hemolysis on blood agar plates. Two cases were polymicrobial infections. In one case with esophageal cancer, clinical symptoms persisted after the start of antimicrobial chemotherapy and the patient died 10 days later associated with complications of esophageal cancer. Leuconostoc lactis, Leuconostoc mesenteroides subsp. dextranicum, and Pediococcus acidilactici wee identified by physiological reactions. These strains were also highly resistant to teicoplanin and fosfomycin, and tolerant to all rested beta-lactams such as benzylpenicillin. This is the first report in Japan to our knowledge on the identification of Leuconostoc spp. and Pediococcus spp. isolated from human infectious diseases.

Aged↗

[The taxonomic distribution, characteristic and susceptibility against antimicrobial agents of methicillin-resistant staphylococci isolated from blood].

The species, mecA gene, beta-lactamase activity, the ability of slime formation and drug susceptibilities of 386 strains of staphylococci which were isolated from blood in our laboratory were studied. The coagulase typing of each strains identified as S. aureus was also determined. These 386 strains consisted of various species, e.g., S. aureus, S. epidermidis, S. capitis, S. caprae, S. hominis, S. simulans, S. haemolyticus, and S. lugdunensis. mecA (methicillin-resistant gene) was detected in 84 (67.7%) of 124 S. aureus and 195 (75.3%) of 259 CNS, but there was no statistical difference. However mecA positive rate was higher in S. epidermidis and S. caprae, lower in S. hominis compared with S. aureus, S. lugdunensis having mecA has not been reported, but one of two our S. lugdunensis strains had mecA. The positive rate (77.4%: 65/84) of beta-lactamase of methicillin-resistant Staphylococcus aureus (MRSA) was lower than that (95.5%: 187/195) of methicillin-resistant coagulase-negative staphylococci (MRCNS). Concerning the ability of slime formation, CNS had higher positive rate, especially in case of MRCNS (46.2%: 84/182) than S. aureus. On the other hand, MRCNS showed a tendency to be less resistant to some antimicrobials than MRSA. Especially against cephalothin (CET), the resistant rate of MRSA and MRCNS were 86.9% and 5.6%, respectively. Among the main species of MRCNS, S. capitis and S. caprae were more resistant than S. epidermidis and S. simulans to cefaclor (CCL), cefmetazole (CMZ), flomoxef (FMOX) and fosfomycin (FOM). Hereafter, one should be careful about infectious disease caused by CNS especially in immuno-compromised host, because many species of CNS have higher positive rate of beta-lactamase and slime formation than S. aureus beside mecA.

Aminoglycosides↗

[Report of questionnaire about enterohemorrhagic Escherichia coli cases caused in the area including Sakai City in 1996].

Many outbreaks of EHEC O157 were occurred in Japan in 1996. There was the biggest outbreak involving approximately 6000 victims in area of Sakai City. We conducted a questionnaire survey on the EHEC infection to the physicians who treated these patients and the microbiologists. For physicians we asked mainly clinical manifestation of patients they observed and treatment they did and their outcome (Answer A). Microbiologists were also required to answer the opinion about treatment (Answer B). Eighty-five of physicians replied about their 222 patients and 209 of microbiologists answered their opinions. In this report we summarized the data and obtained following results: 1) We observed higher frequency of HUS/TTA complication in female than male. 2) Higher HUS/TTP complication was accompanied with patients complained symptoms of fever, bloody diarrhea, and general fatigue. 3) The most patients (96%) were received antimicrobial agents, especially fosfomycin (84%), followed by new quinolon (17.8%) and Cefems (12.2%). Later starting of treatment with antimicrobial agents than 7 days from onset of symptoms increased HUS/TTP complication. 4) Anti-diarrhoeal agents seems to be one of risk factors for HUS/TTP complications.

Adolescent↗