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1.
Clin Microbiol Infect ; 18(9): E347-54, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22738232

ABSTRACT

Antibiotic-resistant bacteria have emerged due to the selective pressure of antimicrobial use in humans and animals. Water plays an important role in dissemination of these organisms among humans, animals and the environment. We studied the antibiotic resistance patterns among 493 Escherichia coli isolates from different aquatic environmental sources collected from October 2008 to May 2009 in León, Nicaragua. High levels of antibiotic resistance were found in E. coli isolates in hospital sewage water and in eight of 87 well-water samples. Among the resistant isolates from the hospital sewage, ampicillin, chloramphenicol, ciprofloxacin, nalidixic acid, trimethoprim-sulphamethoxazole was the most common multi-resistance profile. Among the resistant isolates from the wells, 19% were resistant to ampicillin, ceftazidime, ceftriaxone, cefotaxime, chloramphenicol, ciprofloxacin, gentamicin, nalidixic acid and trimethoprim-sulphamethoxazole. E. coli producing ESBL and harbouring bla(CTX-M) genes were detected in one of the hospital sewage samples and in 26% of the resistant isolates from the well-water samples. The bla(CTX-M-9) group was more prevalent in E. coli isolates from the hospital sewage samples and the bla(CTX-M-1) group was more prevalent in the well-water samples.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Escherichia coli/drug effects , Escherichia coli/isolation & purification , Sewage/microbiology , Wastewater/microbiology , Bacterial Proteins/genetics , Drinking Water/microbiology , Escherichia coli/classification , Escherichia coli/genetics , Hospitals , Humans , Microbial Sensitivity Tests , Nicaragua , Random Amplified Polymorphic DNA Technique , Water Microbiology , beta-Lactamases/genetics
2.
Eur J Clin Microbiol Infect Dis ; 29(6): 715-25, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20383551

ABSTRACT

Correlation between antibody response and clinical outcome in Staphylococcus aureus bacteremia has yielded conflicting results. Immunization schedules have failed in clinical trials. Is the humoral response toward S. aureus of protective nature? A prospective study was performed in patients with invasive S. aureus (ISA) infections during the period 2003-2005. The antibody levels were determined at the beginning and at the end of treatment and one month later (n = 96, n = 71, and n = 51, respectively). As controls, 115 healthy individuals were used. A quantitative enzyme-linked immunosorbent assay (ELISA) against eight purified antigens was performed. Bacterial isolates were grouped as to the production of alpha-toxin, agr type, and pulsed-field gel electrophoresis (PFGE) type. Large variations were seen in the antibody levels. The levels in the second sample were the highest. A correlation between agr group, PFGE group, alpha-toxin production, and initial antibody levels was observed. Patients with fatal outcome displayed lower initial antibody levels to all antigens and significantly so in regard to teichoic acid, lipase, enterotoxin A, and scalded skin syndrome toxin. In episodes with complicated bacteremia, initial significantly low levels to teichoic acid and lipase were registered. Low initial antibody levels against several antigens were associated with increased mortality and complicated bacteremia in invasive S. aureus infections. Bacterial properties, strain, and toxin production affected the antibody response.


Subject(s)
Antibodies, Bacterial/blood , Staphylococcal Infections/immunology , Staphylococcus aureus/immunology , Aged , Antibody Formation , Antigens, Bacterial , Bacterial Proteins/biosynthesis , Bacterial Proteins/immunology , Bacterial Toxins/biosynthesis , Bacterial Toxins/immunology , Bacterial Typing Techniques , DNA Fingerprinting , Electrophoresis, Gel, Pulsed-Field , Enzyme-Linked Immunosorbent Assay/methods , Hemolysin Proteins/biosynthesis , Hemolysin Proteins/immunology , Humans , Middle Aged , Prospective Studies , Staphylococcal Infections/mortality , Staphylococcal Infections/pathology , Staphylococcus aureus/classification , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/pathogenicity , Trans-Activators/biosynthesis , Trans-Activators/immunology , Virulence Factors/biosynthesis , Virulence Factors/immunology
3.
Eur J Clin Microbiol Infect Dis ; 28(9): 1095-103, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19437050

ABSTRACT

The purpose of this paper is to investigate the rate of translocation of Escherichia coli strains in different experimental/animal models. Four proficient translocating E. coli strains isolated from mesenteric lymph nodes (MLNs) and/or the blood of rats (strains KIC-1 and KIC-2), from a fatal case of pancreatitis (HMLN-1) and from pigs (PC-1 isolated in this study) were tested for their ability to translocate across two host species and the Caco-2 cell line as a model of the human gut epithelium. HMLN-1 was found in the MLNs of all 15 pigs tested. This strain, however, did not translocate in any rats and only colonised the caecum of four rats in small numbers. HMLN-1 and PC-1 were the dominant translocating strains in Caco-2 cells compared to KIC-1 and KIC-2, which were found to translocate at a lower rate in pigs and in Caco-2 cells. The rate of translocation of PC-1 in rats was also very low compared to KIC-1 and KIC-2. We suggest that, in studies aiming to investigate the mechanism of translocation of E. coli strains isolated from humans, rats may not be an appropriate animal model and that the Caco-2 cells or pigs are more suitable in vitro and in vivo models, respectively.


Subject(s)
Bacterial Translocation , Escherichia coli Infections/microbiology , Escherichia coli Infections/veterinary , Escherichia coli/pathogenicity , Animals , Bacterial Typing Techniques , Blood/microbiology , Caco-2 Cells , Cluster Analysis , Colony Count, Microbial , Disease Models, Animal , Escherichia coli/classification , Escherichia coli/isolation & purification , Female , Gastrointestinal Tract/microbiology , Humans , Lymph Nodes/microbiology , Male , Rats , Swine
4.
J Appl Microbiol ; 104(5): 1449-57, 2008 May.
Article in English | MEDLINE | ID: mdl-18194253

ABSTRACT

AIMS: To investigate the hypothesis that amoeba may comprise a significant environmental reservoir for Aeromonas, Acanthamoeba-Aeromonas interaction experiments were performed. METHODS AND RESULTS: Acanthamoeba were grown in monoculture and co-cultures with three different species of Aeromonas. Survival, invasion and viable but nonculturable state experiments were performed. We showed that at a low initial bacterial cell density, growth of Aeromonas spp. was inhibited by Acanthamoeba castellanii, while A. castellanii growth was unaffected. In contrast, a high initial bacterial cell density, Aeromonas hydrophila AEW44 and Aeromonas veronii biovar sobria AEW104 suppressed the growth of A. castellanii. Fluorescent and phase-contrast microscopic observations of GFP tagged Aer. hydrophila AEW44 demonstrated that the bacterial cells aggregated on A. castellanii cells after 15 min of incubation and internalized. Aeromonas hydrophila AEW44 cells were found to be actively moving. Interestingly, Aer. hydrophila AEW44 cells shifted more rapidly to a viable but nonculturable form when co-cultured with A. castellanii than in monoculture. CONCLUSIONS: We demonstrated that Aeromonas spp. are able to interact with and to infect the protozoan A. castellanii under laboratory conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Free-living amoeba might play a role as reservoir for Aeromonas, and thus may increase the transmission of Aeromonas by acting as a vehicle.


Subject(s)
Acanthamoeba/microbiology , Aeromonas/physiology , Water Microbiology , Animals , Disease Reservoirs , Fish Diseases/transmission , Gram-Negative Bacterial Infections/transmission , Microscopy, Fluorescence , Microscopy, Phase-Contrast
5.
Appl Environ Microbiol ; 74(5): 1350-6, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18192406

ABSTRACT

The molecular structure and transferability of Tn1546 in 143 vancomycin-resistant Enterococcus faecium (VREF) isolates obtained from patients (n = 49), surface water (n = 28), and urban and hospital sewage (n = 66) in Tehran, Iran, were investigated. Molecular characterization of Tn1546 elements in vanA VREF was performed using a combination of restriction fragment length polymorphism analysis and DNA sequencing of the internal PCR fragments of vanA transposons. Long-PCR amplification showed that the molecular size of Tn1546 elements varied from 10.8 to 12.8 kb. The molecular analysis of Tn1546 showed that 45 isolates (31.5%) harbored a deletion/mutation upstream from nucleotide 170. No horizontal transfer of Tn1546 was observed following filter-mating conjugation with these isolates. Nevertheless, the rates of transferability for other isolates were 10(-5) to 10(-6) per donor. Insertion sequences IS1216V and IS1542 were present in 103 (72%) and 138 (96.5%) of the isolates, respectively. The molecular analysis of Tn1546 elements resulted in three genomic organizations. The genomic organization lineage 1 was dominated by the isolates from clinical samples (3.4%), lineage 2 was dominated mostly by sewage isolates (24.5%), and lineage 3 contained isolates obtained from all sources (72.1%). The genetic diversity determined using pulsed-field gel electrophoresis (PFGE) revealed a single E. faecium clone, designated 44, which was common to the samples obtained from clinical specimens and hospital and municipal sewage. Furthermore, the results suggest that lineage 3 Tn1546 was highly disseminated among our enterococcal isolates in different PFGE patterns.


Subject(s)
DNA Transposable Elements/genetics , Enterococcus faecium/genetics , Inpatients , Sewage/microbiology , Vancomycin Resistance/genetics , Water Microbiology , Base Sequence , Cluster Analysis , Electrophoresis, Gel, Pulsed-Field , Genome Components , Humans , Iran , Molecular Sequence Data , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA
6.
Allergy ; 61(8): 1002-8, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16867055

ABSTRACT

BACKGROUND: The opportunistic yeast Malassezia is a trigger factor in atopic eczema (AE). Around 30-80% of patients with AE have an IgE and/or T-cell reactivity to the yeast. Several IgE-binding components have been identified in Malassezia extracts and 11 allergens have been cloned and sequenced. The pH of the skin surface in patients with AE is higher than that of normal healthy skin. We here investigate whether different pH conditions mimicking those of AE skin and healthy skin can influence the production and release of Malassezia allergens. METHODS: Malassezia sympodialis (ATCC strain 42132) was cultured in Dixon broth at pH 6.1 to 5.0 for 1-15 days. Culture supernatants were analysed for the presence of IgE-binding components by immunoblotting. The M. sympodialis cells were analysed for allergen expression and production with immunocytochemistry and quantitative polymerase chain reaction. RESULTS: We found that M. sympodialis cells produce, express and release allergens to a greater extent when cultured at the higher pH. This was particularly true of a 67-kDa major allergen designated Mala s 12. CONCLUSIONS: The data suggest that the skin barrier in AE patients provides an environment that can enhance the release of allergens from M. sympodialis, which can contribute to the inflammation.


Subject(s)
Allergens/immunology , Antigens, Fungal/immunology , Dermatitis, Atopic/immunology , Dermatomycoses/immunology , Malassezia/immunology , Skin/immunology , Allergens/biosynthesis , Antigens, Fungal/biosynthesis , Dermatitis, Atopic/microbiology , Dermatomycoses/microbiology , Humans , Hydrogen-Ion Concentration , Immunoglobulin E/immunology , Malassezia/growth & development , T-Lymphocytes/immunology , T-Lymphocytes/microbiology
7.
Epidemiol Infect ; 134(5): 985-9, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16650334

ABSTRACT

During recent years a pandemic clone of Vibrio parahaemolyticus has emerged. Isolates of this clone are distributed among several serotypes, but are genotypically related. In the present study, a phenotyping method (biochemical fingerprinting) was used to characterize pandemic and non-pandemic isolates belonging to V. parahaemolyticus. It was found that the pandemic isolates showed a high level of phenotypic homogeneity and a majority of the pandemic isolates belonged to the same biochemical phenotype, whereas non-pandemic V. parahemolyticus isolates were more heterogeneous. In conclusion, biochemical fingerprinting of V. parahaemolyticus can be used as a first screening method to differentiate between pandemic and non-pandemic isolates of V. parahaemolyticus.


Subject(s)
Bacterial Typing Techniques , Vibrio parahaemolyticus/classification , Communicable Diseases, Emerging/microbiology , Food Microbiology , Humans , India/epidemiology , Phenotype , Seafood , Vibrio Infections/epidemiology , Vibrio Infections/microbiology , Vibrio parahaemolyticus/isolation & purification
8.
J Microbiol Methods ; 63(3): 229-38, 2005 Dec.
Article in English | MEDLINE | ID: mdl-15979748

ABSTRACT

Long-time storage of faecal samples is necessary for investigations of intestinal microfloras. The aim of the present study was to evaluate how the viability and the composition of the Escherichia coli flora are affected in faecal samples during different storage conditions. Four fresh faecal samples (two from calves and two from infants) were divided into sub-samples and stored in four different ways: with and without addition of glycerol broth at -20 degrees C and at -70 degrees C. The viability and the phenotypic diversity of the E. coli flora in the sub-samples were evaluated after repeated thawings and after storage during 1 year. The samples stored for 1 year without thawing were also kept at room temperature for 5 days and subsequently analysed. According to phenotyping (PhP analysis) of 32 isolates per sample on day 0, all four samples contained two dominating strains of E. coli each, and between one and eight less common strains. Samples that were stored at -70 degrees C in glycerol broth showed equal or even higher bacterial numbers as the original samples, even after repeated thawings, whereas samples stored at -20 degrees C showed a considerably lower survival rate, also with addition of glycerol. Sub-samples containing glycerol broth that were kept at room temperature after storage for 1 year showed a clear increase in the number of viable cells as well as in diversity. The diversities in each sub-sample showed a tendency to decrease after several thawings as well as after storage. Generally, the E. coli populations in samples stored at -20 degrees C were less similar to the population of the original sample than that in samples stored at -70 degrees C. Samples that had been mixed with glycerol broth had an E. coli flora more similar to that in the original sample than those without glycerol broth. Furthermore, the sub-samples that were kept at room temperature after storage for 1 year generally were more similar to the original samples than if they were processed directly. We conclude that for long time storage of faecal samples, storage at -70 degrees C is preferable. If samples have to be thawed repeatedly, addition of glycerol is preferable both for samples stored at -70 degrees C and for samples stored at -20 degrees C. Our data also have indicated that when E. coli isolates from faecal samples are selected for, e.g. analysis of virulence factors, it is necessary to pick several isolates per sample in order to obtain at least one isolate representing the dominating strain(s).


Subject(s)
Escherichia coli/isolation & purification , Feces/microbiology , Animals , Bacteriological Techniques , Cattle , Cryopreservation , Glycerol , Humans , Infant , Phenotype , Time Factors
9.
J Infect ; 50(4): 312-21, 2005 May.
Article in English | MEDLINE | ID: mdl-15845429

ABSTRACT

BACKGROUND: Escherichia coli strains that cause cystitis posses virulence properties that facilitate their colonisation and persistence in the bladder. In Iran, despite the high number of the urinary tract infections, very few studies has been done to determine the role of these virulence properties in the pathogenesis of E. coli cyctitis. PATIENTS AND METHODS: Eighty-seven strains of E. coli, isolated from young adults with cystitis in Shiraz, Iran, were examined for the expression of type 1 and P-fimbriae, mannose resistant haemagglutination, haemolysin production, aerobactin-mediated iron uptake, O:K serotypes, biochemical phenotypes (BPTs) and their antibiotic susceptibility patterns. RESULTS: Seventy-six percent of the strains expressed multiple virulence properties. There was a significant correlation between the presence of aerobactin and the expression of type 1 fimbriae. All P-fimbriated strains produced aerobactin with 50% of them also coexpressing haemolysin. Of the 29 different O:K serotypes identified, 42% belonged to serotypes not commonly found among European serotypes associated with UTI. Strains of O groups 4 and 6 expressed more virulence factors than the others. A high resistance against ampicillin, trimethoprim and cotrimoxasol was observed among the isolates with 53% of the isolates showing multiresistance to these three antibiotics. Certain BPTs were also found among O:K serotypes with some containing strains of the same virulence profile. CONCLUSION: We conclude that certain colonal groups of E. coli are commonly associated with cystitis in young adults in Iran with strains possessing a combination of aerobactin and type 1 fimbriae being the dominant ones and belonging to serotypes not commonly found in Europe. We also conclude that the multiple antibiotic resistant E. coli strains causing cyctitis are highly prevalent in this part of the country.


Subject(s)
Cystitis/etiology , Escherichia coli/pathogenicity , Virulence Factors/analysis , Acute Disease , Adult , Cystitis/microbiology , Escherichia coli/classification , Escherichia coli/drug effects , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged
10.
Acta Vet Scand ; 45(1-2): 27-36, 2004.
Article in English | MEDLINE | ID: mdl-15535084

ABSTRACT

The prevalence of diarrhoea in calves was investigated in 8 dairy farms in Mozambique at 4 occasions during 2 consecutive years. A total of 1241 calves up to 6 months of age were reared in the farms, and 63 (5%) of them had signs of diarrhoea. Two farms had an overall higher prevalence (13% and 21%) of diarrhoea. Faecal samples were collected from all diarrhoeal calves (n = 63) and from 330 healthy calves and analysed for Salmonella species, Campylobacter jejuni and enterotoxigenic Escherichia coli (ETEC). Salmonella spp. was isolated in only 2% of all calves. Campylobacter was isolated in 11% of all calves, irrespective of health condition, and was more frequent (25%) in one of the 2 diarrhoeal farms (p = 0.001). 80% of the isolates were identified as C. jejuni. No ETEC strains were detected among the 55 tested strains from diarrhoeal calves, but 22/55 (40%) strains from diarrhoeal calves and 14/88 (16%) strains from healthy calves carried the K99 adhesin (p = 0.001). 6,757 E. coli isolates were typed with a biochemical fingerprinting method (the PhenePlate) giving the same E. coli diversity in healthy and diarrhoeal calves. Thus it was concluded: i) the overall prevalence of diarrhoea was low, but 2 farms had a higher prevalence that could be due to an outbreak situation, ii) Salmonella did not seem to be associated with diarrhoea, iii) Campylobacter jejuni was common at one of the 2 diarrhoeal farms and iv) ETEC strains were not found, but K99 antigen was more prevalent in E. coli strains from diarrhoeal calves than from healthy, as well as more prevalent in one diarrhoeal farm.


Subject(s)
Cattle Diseases/epidemiology , Cattle Diseases/microbiology , Diarrhea/veterinary , Animals , Animals, Newborn , Campylobacter jejuni/isolation & purification , Case-Control Studies , Cattle , Dairying , Diarrhea/epidemiology , Diarrhea/microbiology , Mozambique/epidemiology , Prevalence
11.
Eur J Clin Microbiol Infect Dis ; 23(2): 98-105, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14735403

ABSTRACT

The aims of this study were to evaluate the serological response to treatment with staphylococcal vaccine in fibromyalgia/chronic fatigue syndrome patients and to explore the relationship between serological response and clinical effect. Twenty-eight patients, half of whom served as controls, were recruited from a 6-month randomised trial in which repeated administration of the staphylococcal toxoid vaccine Staphypan Berna (Berna Biotech, Switzerland) was tested against placebo. Antibody status against extracellular toxins/enzymes, cell-wall components, and enterotoxins was evaluated at baseline and at endpoint. The clinical response to treatment was recorded in rating scales. In the group receiving active treatment, significant serological changes were recorded, whereas no significant changes were found in controls. Treatment led to a significantly increased capacity of serum to neutralise alpha-toxin and a significant increase in serum IgG to alpha-toxin and lipase. Furthermore, the increase in these parameters combined paralleled the improvement in clinical outcome. Thus, the greater the serological response, the greater was the clinical effect. In conclusion, this explorative study has shown that repeated administration of the Staphypan Berna vaccine in patients with fibromyalgia/chronic fatigue syndrome causes a serological response to several staphylococcal antigens, particularly to certain extracellular toxins and enzymes. The results further show that this response is related to the clinical outcome of treatment.


Subject(s)
Antibodies, Bacterial/analysis , Fatigue Syndrome, Chronic/immunology , Fatigue Syndrome, Chronic/therapy , Fibromyalgia/immunology , Fibromyalgia/therapy , Staphylococcal Vaccines/therapeutic use , Adult , Enzyme-Linked Immunosorbent Assay , Fatigue Syndrome, Chronic/complications , Female , Fibromyalgia/complications , Follow-Up Studies , Humans , Immunity/physiology , Male , Middle Aged , Probability , Reference Values , Risk Assessment , Serologic Tests , Statistics, Nonparametric , Treatment Outcome
12.
Rev. méd. Chile ; 131(8): 837-45, ago. 2003.
Article in Spanish | LILACS | ID: lil-356051

ABSTRACT

BACKGROUND: PhP-S48 (Phene Plate Techniques AB), a method based on biochemical phenotypes has been developed and used successfully to typify S enteritidis strains in epidemiological studies. AIM: To identify phenotypes of S enteritidis isolated from eggs, chicken meat and infected humans in Antofagasta during the period 1997-2000. MATERIAL AND METHODS: PhP-S48 and phage typing were used to identify phenotypes of 33 S enteritidis strains, sixteen isolated from poultry and 17 from clinical sources. S enteritidis ATCC17036 was used as control strain. RESULTS: Twelve biochemical phenotypes (BTs) including 4 common (C) and 8 single (S) were identified. BTs C1 y C3 containing 16 and 5 strains, respectively, accounted for 63.6 per cent of the isolates. BT C1 was found in poultry and human sources in the period 1997-2000, and BT C3 was isolated from humans, in the period 1999-2000. Using phage typing, 5 phage types (PT) and 3 strains could be not typed (NTs). PT1 and PT21 were the dominant phage types, with 14 and 13 strains respectively. Strains of PT1 were isolated from poultry and human sources in the period 1997-2000. PT21 was found in poultry samples in the period 1997-1998 and in clinical samples, in the period 1997-1998. Combination of biochemical phenotypes and phage typing divided the strains into 5 phenotypes (BT:PT). Two phenotypes were the most frequently isolated, phenotype C1:1 with 8 isolates found in eggs and humans in 1999, and phenotype C1:21 with 5 strains isolated in 1997-1999. CONCLUSIONS: These results indicate the presence of one persistent and one recently emerged phenotype among S enteritidis in Antofagasta, Chile. PhP-S48 also provided information about a relationship among the strains.


Subject(s)
Humans , Phenotype , Food Microbiology , Poultry Products/microbiology , Salmonella enteritidis/classification , Bacteriophage Typing , Poultry , Chile , Salmonella enteritidis/genetics , Salmonella enteritidis/isolation & purification
13.
J Med Microbiol ; 52(Pt 2): 109-112, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12543915

ABSTRACT

A young female with no identifiable risk factors developed rapid, overwhelming Staphylococcus aureus endocarditis. Despite rapid sterilization of the blood and the mitral valve with optimal antimicrobials, she had persistent septic shock. In order to investigate this, the toxin-producing capacity of the infecting strain and the patient's ability to produce antibodies were determined. The strain produced high levels of both alpha-toxin and staphylococcal enterotoxin A (SEA), whilst the patient responded with modestly high levels of antibodies to alpha-toxin and low-normal levels to SEA. The patient was most probably susceptible to the actions of SEA and developed a toxic-shock-syndrome-like disease that further aggravated her valvular dysfunction. This case illustrates that optimal antimicrobial therapy alone is not sufficient treatment in patients with persistent toxic shock and that there is a need to evaluate immunomodulatory strategies in such patients.


Subject(s)
Endocarditis, Bacterial/microbiology , Enterotoxins/physiology , Shock, Septic/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/pathogenicity , Adult , Anti-Bacterial Agents/therapeutic use , Antibodies, Bacterial/biosynthesis , Bacterial Toxins/biosynthesis , Bacterial Toxins/immunology , Endocarditis, Bacterial/drug therapy , Endocarditis, Bacterial/immunology , Enterotoxins/biosynthesis , Enterotoxins/immunology , Fatal Outcome , Female , Hemolysin Proteins/biosynthesis , Hemolysin Proteins/immunology , Humans , Shock, Septic/immunology , Staphylococcal Infections/drug therapy , Staphylococcal Infections/immunology , Staphylococcus aureus/immunology , Staphylococcus aureus/metabolism
14.
J Clin Microbiol ; 38(10): 3785-90, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11015403

ABSTRACT

Aeromonads are causative agents of a number of human infections. Even though aeromonads have been isolated from patients suffering from diarrhea, their etiological role in gastroenteritis is unclear. In spite of a number of virulence factors produced by Aeromonas species, their association with diarrhea has not been clearly linked. Recently, we have characterized a heat-labile cytotonic enterotoxin (Alt), a heat-stable cytotonic enterotoxin (Ast), and a cytotoxic enterotoxin (Act) from a diarrheal isolate of Aeromonas hydrophila. Alt and Ast are novel enterotoxins which are not related to cholera toxin; Act is aerolysin related and has hemolytic, cytotoxic, and enterotoxic activities. We studied the distribution of the alt, ast, and act enterotoxin genes in 115 of 125 aeromonads isolated from 1, 735 children with diarrhea, in all 27 aeromonads isolated from 830 control children (P = 7 x 10(-4) for comparison of rates of isolation of aeromonads from cases versus those from controls), and in 120 randomly selected aeromonads from different components of surface water in Bangladesh. Aeromonas isolates which were positive only for the presence of the alt gene had similar distributions in the three sources; the number of isolates positive only for the presence of the ast gene was significantly higher for the environmental samples than for samples from diarrheal children; and isolates positive only for the presence of the act gene were not found in any of the three sources. Importantly, the number of isolates positive for both the alt and ast genes was significantly higher for diarrheal children than for control children and the environment. Thus, this is the first study to indicate that the products of both the alt and ast genes may synergistically act to induce severe diarrhea. In 26 patients, Aeromonas spp. were isolated as the sole enteropathogen. Analysis of clinical data from 11 of these patients suggested that isolates positive for both the alt and ast genes were associated with watery diarrhea but that isolates positive only for the alt gene were associated with loose stools. Most of the isolates from the three sources could be classified into seven phenospecies and eight hybridization groups. For the first time, Aeromonas eucrenophila was isolated from two children, one with diarrhea and another without diarrhea.


Subject(s)
Aeromonas/genetics , Aeromonas/isolation & purification , Diarrhea/microbiology , Enterotoxins/genetics , Water Microbiology , Aeromonas hydrophila/genetics , Aeromonas hydrophila/isolation & purification , Bangladesh , Child, Preschool , DNA Probes , Feces/microbiology , Female , Gastroenteritis/microbiology , Humans , Infant , Male , Rectum/microbiology , Reference Values , Serotyping
15.
Int J Antimicrob Agents ; 14(4): 337-42, 2000 May.
Article in English | MEDLINE | ID: mdl-10794956

ABSTRACT

The objectives of the present study are to generate knowledge of the ecology and epidemiology of enterococci in the food chain by studying the following: (1) the population structure (in measures of abundance, number of vancomycin resistant strains, antibiotic resistance patterns, diversity, and stability) among enterococcal populations in different geographical regions and in different links of the food chain (2) possible transmission of strains through the food chain and between hospital environments and the food chain (3) the association between vancomycin resistance and individual strains of enterococci and (4) the diversity of the drug resistance genes in enterococci. So far, 1578 samples have been collected from different countries within the EU (Sweden, Denmark, UK and Spain), and from different habitats (pig farms, carcasses in slaughter houses, soil, manure, water, sewage, and humans). Total and vancomycin resistant enterococcal populations in each sample have been enumerated and more than 12000 isolates have been characterised by phenotyping. Representative isolates are further species identified and characterised by genotyping and MIC determination and from antibiotic resistant isolates the resistance genes are characterised.


Subject(s)
Drug Resistance, Microbial , Enterococcus/drug effects , Food Microbiology , Gram-Positive Bacterial Infections/epidemiology , Vancomycin Resistance , Animals , Enterococcus/classification , Europe , Government Programs , Humans , International Cooperation , Microbial Sensitivity Tests , Phenotype , Research
17.
J Med Microbiol ; 49(2): 177-185, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10670569

ABSTRACT

Recent case-control studies in Bangladesh showed a high prevalence of enteropathogenic Escherichia coli (EPEC) strains (identified by DNA probes for virulence genes) associated with childhood diarrhoea. However, the clonal status of these strains is not known. A total of 94 EPEC isolates from 80 children with diarrhoea and 14 healthy matched controls isolated during 1991-1992 and 1993-1994 was characterised by serogrouping, enterobacterial repetitive intergenic consensus sequence PCR, and by a biochemical fingerprinting method (the phene plate or PhP system). Twelve O serogroups were found with O114 (n = 19) and O127 (n = 23) being the dominant serogroups. Most strains of O114 belonged to the same PhP/PCR types. Strains of O127 contained 16 that produced cytolethal distending toxin (CDT) and seven that did not; both were found among patients as well as controls. Results of PCR and PhP typing showed that CDT-positive strains belonged to the same clonal group and were related to one of the two PhP/PCR types of CDT-negative O127 strains. Thirty-one EPEC strains were O non-typable and 21 strains belonged to other less prevalent serogroups. These strains belonged to diverse PhP/PCR types and did not show any similarity to the strains of two major serogroups, O114 and O127. The results suggest that two clonal groups of EPEC strains are predominantly associated with childhood diarrhoea in Bangladesh.


Subject(s)
Diarrhea/microbiology , Escherichia coli Infections/microbiology , Escherichia coli/classification , Escherichia coli/genetics , Bacterial Typing Techniques , Bangladesh , Case-Control Studies , Child, Preschool , DNA Fingerprinting/methods , DNA, Bacterial/analysis , Diarrhea/epidemiology , Escherichia coli/isolation & purification , Escherichia coli Infections/epidemiology , Humans , Infant , Infant, Newborn , Polymerase Chain Reaction/methods , Prevalence , Serotyping
18.
J Infect Dis ; 181(2): 774-8, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10669375

ABSTRACT

Escherichia coli FimH adhesin mediates binding to the bladder mucosa. In mice, a FimH vaccine protects against bacterial challenge. In this study, 4 monkeys were inoculated with 100 microgram of FimCH adhesin-chaperone complex mixed with MF59 adjuvant, and 4 monkeys were given adjuvant only intramuscularly. After 2 doses (day 0 and week 4), a booster at 48 weeks elicited a strong IgG antibody response to FimH in the vaccinated monkeys. All 8 monkeys were challenged with 1 mL of 108 E. coli cystitis isolate NU14. Three of the 4 vaccinated monkeys were protected from bacteruria and pyuria; all control monkeys were infected. These findings suggest that a vaccine based on the FimH adhesin of E. coli type 1 pili may have utility in preventing cystitis in humans.


Subject(s)
Adhesins, Bacterial/immunology , Adhesins, Escherichia coli , Antibodies, Bacterial/blood , Bacterial Vaccines/immunology , Escherichia coli Infections/prevention & control , Escherichia coli/immunology , Fimbriae Proteins , Urinary Tract Infections/prevention & control , Adhesins, Bacterial/administration & dosage , Animals , Bacterial Vaccines/administration & dosage , Escherichia coli/growth & development , Escherichia coli/pathogenicity , Escherichia coli Infections/microbiology , Feces/microbiology , Humans , Macaca fascicularis , Stomach/microbiology , Urinary Bladder/microbiology , Urinary Tract Infections/microbiology , Vaccination
19.
Clin Diagn Lab Immunol ; 7(1): 14-20, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10618270

ABSTRACT

We analyzed the serum antibody responses against two Staphylococcus aureus fibrinogen binding proteins, the cell-bound clumping factor (Clf) and an extracellular fibrinogen binding protein (Efb). The material consisted of 105 consecutive serum samples from 41 patients suffering from S. aureus septicemia and 72 serum samples from healthy individuals. An enzyme-linked immunosorbent assay (ELISA) was developed. Healthy individuals showed variable levels of antibodies against the studied antigens, and cutoff levels (upper 95th percentile) against these antigens were determined. No correlation was seen between serum antibody levels against Clf and Efb. In acute-phase samples 27% of patients showed positive antibody levels against Clf and 10% showed positive levels against Efb, while in convalescent-phase samples 63% (26 of 41) showed a positive serology against Clf and 49% (20 of 41) showed a positive serology against Efb. Antibody levels against Efb were significantly lower in the acute-phase sera than in sera from healthy individuals (P = 0. 002). An antibody response against Clf was most frequent in patients suffering from osteitis plus septic arthritis and from endocarditis (80% positive). The antibody response against Efb appeared to develop later in the course of disease. A possible biological effect of measured antibodies was demonstrated with the help of an inhibition ELISA, in which both high-titer and low-titer sera inhibited the binding of bacteria to fibrinogen. In conclusion, we have demonstrated in vivo production of S. aureus fibrinogen binding proteins during deep S. aureus infections and a possible diagnostic and prophylactic role of the corresponding serum antibodies in such infections.


Subject(s)
Coagulase/immunology , Sepsis/immunology , Staphylococcal Infections/immunology , Staphylococcus aureus/immunology , Adolescent , Adult , Aged , Antibodies/immunology , Antigens/immunology , Arthritis, Infectious/immunology , Bacterial Proteins/immunology , Carrier Proteins/immunology , Endocarditis/immunology , Enzyme-Linked Immunosorbent Assay/methods , Humans , Middle Aged , Osteitis/immunology , Sweden
20.
J Appl Microbiol ; 87(4): 564-73, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10583685

ABSTRACT

The effect of a dietary supplementation of zinc oxide (ZnO) on the stability of the intestinal flora and on the composition of coliforms in weaned pigs was investigated. Faecal floras were characterized by their metabolic activities and fermentative capacity (FC) using the Phene Plate generalized microplate. Coliforms were characterized by conventional enumeration and by the Phene Plate-RS plates. The latter measured FC, phenotypic diversity, persistence of each coliform strain in piglets, and similarity among the coliform populations within groups. From weaning onwards, the control pigs (n = 5) were fed a basal diet ad libitum, while experimental pigs (n = 5) were given the same food supplemented with 2500 ppm ZnO. Metabolic fingerprinting of faecal floras indicated marked differences between the composition of floras of treated and control pigs during the first 2 weeks post-weaning. The FC of faecal flora in both groups decreased as pigs aged, but it was significantly (P

Subject(s)
Dietary Supplements , Enterobacteriaceae/growth & development , Intestines/microbiology , Zinc Oxide/pharmacology , Animals , Animals, Newborn , Enterobacteriaceae/classification , Feces/chemistry , Feces/microbiology , Phenotype , Swine , Weaning , Zinc Oxide/metabolism
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