Characterization of Pasteurella spp. strains isolated from human infections

This report describes the distribution of species and capsular groups in a collection of 143 strains of Pasteurella recovered from human patients. The organism isolated most frequently was Pasteurella multocida subsp. multocida. As in animals, most of the group A strains were recovered from the respiratory tract. The distribution of species in relation to the animal source suggests that P. multocida subsp. multocida is more infective than other Pasteurella species or subspecies for man.     

Structural and pathogenic properties of Aeromonas schubertii.

We investigated the phenotypic, structural, and pathogenic properties of 11 Aeromonas schubertii strains recovered from extraintestinal sites. Most A. schubertii strains were autoagglutination positive, possessed a high surface charge but low hydrophobicity, and fell into one or two biogroups on the basis of carbon substrate utilization patterns. Fatty acid methyl ester analysis of A. schubertii revealed this species to contain a relatively high percentage of branched fatty acids (i-13:0, i-15:0, i-17:1, i-17:0) compared with A. hydrophila. Immunologic and biochemical analysis of the lipopolysaccharides of A. schubertii strains allowed for two groups to be distinguished, namely, (i) a collection of six strains belonging to serogroup O:11 that possessed a characteristic homogeneous O polysaccharide side chain profile by silver staining and immunoblotting techniques and (ii) a second antigenically diverse group (five strains) that either exhibited a heterogeneous side chain profile or were side chain deficient. A, schubertii O:11 strains were all found to contain a 55-kDa major protein associated with the outer membrane fraction which was glycine-hydrochloride extractable; non-O:11 strains did not harbor a similar protein molecule. Screening of A. schubertii strains for reputed virulence factors indicated (i) that slightly more than half of the isolates produce an apparent contact-dependent hemolysin that is not cell associated or released extracellularly, (ii) a potent cytotoxin active against HEp-2 cells that is devoid of hemolytic activity, and (iii) lack of enterotoxigeniclike activity as determined by suckling mouse assays. All A. schubertii strains were pathogenic for mice as determined by 50% lethal dose assays, although no single factor correlated with mouse pathogenicity.     

Characterisation of Eubacterium-like strains isolated from oral infections.

The genus Eubacterium currently includes a heterogeneous group of gram-positive, non-spore-forming anaerobic bacilli, many of which are slow growing, fastidious and generally unreactive in biochemical tests. As a consequence, cultivation and identification of isolates are difficult and the taxonomy of the group remains indifferent. In this study, 105 isolates from odontogenic infections, infections associated with dental implants or saliva from healthy subjects and provisionally assigned to the genus Eubacterium were subjected to phenotypic and genotypic analysis. Ninety-one of the isolates were identified as belonging to one of 14 previously described species: Atopobium parvulum (5 isolates), A. rimae (29), Bulleidia extructa (2), Cryptobacterium curtum (1), Dialister pneumosintes (1), Eubacterium saburreum (2), E. sulci (8), E. yurii subsp. yurii (1), Filifactor alocis (3), Lactobacillus uli (1), Mogibacterium timidum (13), M. vescum (6), Pseudoramibacter alactolyticus (6) and Slackia exigua (13). The remaining 14 isolates did not correspond to existing species. This study confirms the diversity of organisms provisionally assigned to the genus Eubacterium by conventional identification methods. This group of organisms is frequently isolated from oral infections but their role in the aetiology of these conditions has yet to be determined.     

Virulence properties and enterotoxin production of Aeromonas strains isolated from fish.

The biological activities in vivo and in vitro of 59 motile Aeromonas spp. isolated from fish and water tanks were simultaneously analyzed in poikilothermic and homoiothermic systems. A total of 64.3% of the isolates tested were pathogenic for fish, and 62% of Aeromonas hydrophila and A. sobria isolates either virulent or nonvirulent for fish were enterotoxigenic. Although the majority of the strains were proteolytic and amylolytic and produced DNase, other activities, such as elastase and staphylolysis, were only present in A. hydrophila. Most of the strains (96%) produced hemolysins, and 68% had agglutinating capacity, but neither isolates pathogenic for fish nor enterotoxigenic isolates showed specificity for trout or human erythrocytes, respectively. The production of siderophores, agglutination in acriflavine, and precipitation after boiling were found not to be useful tests for screening virulent strains. Although statistical analysis revealed a significant relationship between virulence for fish and positive results for arabinose and sucrose fermentations, elastase, and hemolysis of human erythrocytes, only lysine decarboxylase showed a significant positive relationship with enterotoxigenicity. Using extracellular products from representative Aeromonas strains with different virulence markers and belonging to distinct O serogroups, we demonstrated a lack of correlation between cytotoxicity for fish and homoiothermic cell lines and pathogenicity. The extracellular products from selected pathogenic A. hydrophila strains were lethal for rainbow trout and displayed proteolytic, hemolytic, and cytotoxic activities which were simultaneously lost after heat treatment. The findings reported here indicate that it is not possible to establish a common and single mechanism involved in the invasion of Aeromonas spp. in poikilothermic and homoiothermic hosts.     

Haemolysin occurrence among Aeromonas hydrophila, Aeromonas caviae and Aeromonas sobria strains isolated from different aquatic ecosystems

A total of 909 Aeromonas spp. isolates from different aquatic ecosystems were tested for haemolysin production by both sheep and horse blood agar-plate assays and by rabbit erythrocytes in broth assay. A comparison of these different methods was undertaken in order to appreciate their capacity to evaluate the haemolytic activity of Aeromonas spp. isolated from aquatic ecosystems. The haemolytic activity was associated particularly with A. hydrophila and A. sobria (about 95% of strains), whereas A. caviae did not produce haemolysin (about 95% of strains). A method suitable for use in routine diagnostic microbiology laboratories is proposed for quantifying both groups of A. hydrophila/A. sobria and A. caviae in environmental water.     

Aeromonas schubertii, a new mannitol-negative species found in human clinical specimens.

In 1983 the vernacular name Enteric Group 501 was coined for a group of strains that had been referred to our laboratory as "possible Vibrio damsela that does not require NaCl for growth." By DNA-DNA hybridization (hydroxyapatite method, 32P, 60 and 75 degrees C), six strains of Enteric Group 501 were closely related to the labeled strain 2446-81 (70 to 95% at 60 degrees C and 71 to 93% at 75 degrees C; 0 to 1% divergence). Type strains of all Aeromonas species and reference strains of six other Aeromonas DNA hybridization groups were 26 to 42% related (60 degrees C) to strain 2446-81, but type strains of 27 Vibrio and Photobacterium species, including V. damsela, were 0 to 1% (75 degrees C) related. We propose the name Aeromonas schubertii for the highly related group of seven strains formerly known as Enteric Group 501. The type strain is designated as ATCC 43700 (CDC 2446-81). Strains of A. schubertii grew well at 36 degrees C and had positive reactions at this temperature for methyl red, Voges-Proskauer (1% NaCl, Coblentz method), lysine decarboxylase, arginine dihydrolase, motility, lipase, DNase, nitrate reduction to nitrite, oxidase, and growth in nutrient broth with 0 and 1% NaCl. There was no growth in 6% NaCl or on thiosulfate-citrate-bile salts-sucrose agar. The following sugars were fermented: D-glucose, D-galactose, maltose, D-mannose, and trehalose. The following sugars were not fermented: adonitol, L-arabinose, D-arabitol, cellobiose, dulcitol, erythritol, myo-inositol, lactose, D-mannitol, melibiose, alpha-CH3-D-glucoside, raffinose, L-rhamnose, salicin, D-sorbitol, sucrose, and D-xylose. Esculin was not hydrolyzed, and the string test was negative. The mannitol-negative reaction differtiates A. schubertii from other Aeromonas species. The antibiogram of this organism is typical of other Aeromonas strains (resistance to ampicillin and carbenicillin and susceptibility to most other agents). A. schubertii strains have been isolated from abscesses (two strains), wound (one), skin (one), pleural fluid (one), and blood (two). The two blood isolates suggest clinical significance typical of other Aeromonas species , but further information is needed on this group.     

Characterization of the Aeromonas hydrophila group isolated from retail foods of animal origin.

During a recent survey of retail fresh foods of animal origin (fish and seafood, raw milk, poultry, and red meats) for organisms of the Aeromonas hydrophila group, we isolated representative strains from the various foods. In this study, we sought to characterize these isolates for biochemical properties and virulence-associated factors and to compare the food isolates with clinical isolates. We identified all food and clinical isolates as A. hydrophila and found that all isolates were typical in their biochemical reactions. Examination of the isolates for various virulence-associated factors indicated that most food and clinical isolates were serum resistant, beta-hemolytic, cytotoxin positive (against Y1 adrenal cells), hemagglutinin positive, Congo red positive, elastase positive, and staphylolysin positive. Mouse 50% lethal doses were log10 8 to 9 CFU for most isolates. All isolates had biotypes identical to those of enterotoxin-positive strains. The public health significance of these organisms in foods is not known at present, although their widespread occurrence and ability to grow competitively in foods kept at 5 degrees C represents a potential hazard.     

Fimbrial serotypes of Escherichia coli strains isolated from extra-intestinal infections

Strains of Escherichia coli isolated from urinary tract infections and meningitis were characterised by their O:K serotype, haemolysin production, mannose-resistant haemagglutination, and the serotype of the P-fimbriae. The P-fimbriae of 71% of the mannose-resistant haemagglutination-positive strains from urinary tract infection and meningitis could be determined with specific monoclonal antibodies. Many strains expressed multiple P-fimbriae serotypes. The serotypes of P-fimbriae found most frequently among mannose-resistant haemagglutination-positive E. coli from urinary tract infections were the F11, F7 and F8 fimbriae, and among meningitic strains, F11, F8 and F9 fimbriae. The expression of certain F-serotypes did not correlate with O:K antigens.     

Characterization of orthopoxviruses isolated from feline infections in Britain

Summary The biological properties and genomes of orthopoxviruses isolated from cats in Britain were compared with strains of cowpox virus isolated from cows and their handlers.All the isolates tested produced haemorrhagic pocks and A-type inclusions on the CAM, but did not produce pocks above 40°C. Thus the feline isolates behaved as typical strains of cowpox virus. Differences were found in the heat resistance of the virions and in the character of the A-type inclusion which did not correlate with the host from which the viruses were isolated. Analysis of the genomes with a variety of restriction endonucleases showed very close relationship between all the isolates and also failed to separate feline isolates from cowpox virus. However again minor differences, which may prove to be of epidemiological value were detected.We conclude that the orthopoxvirus currently isolated from domestic cats in Britain is cowpox virus and that there is no evidence that a feline variant or subspecies circulates in Britain.     

Characterization of Borrelia burgdorferi strains isolated from Ixodes pacificus ticks in California.

In a survey of 1,714 adult Ixodes pacificus ticks collected in northern California, 24 (1.4%) were found to be infected with spirochetes that reacted with an anti-Borrelia burgdorferi polyvalent conjugate in direct immunofluorescence tests. Eleven isolates of B. burgdorferi from these ticks were characterized by monoclonal antibody, polyacrylamide gel electrophoresis, and Western blot (immunoblot) analyses. Ten of the isolates had molecular and antigenic characteristics similar to those of other U.S. isolates. One strain, cloned by limiting-dilution techniques, was different from any previously reported U.S. strain, but similar to reported European strains. The cloned strain, DN127-Cl9-2, did not react with monoclonal antibodies to Osp A and Osp B major proteins found in most of the U.S. strains. It exhibited an abundant protein with an apparent molecular weight of 25,000.     

Characteristics of cysteine-requiring strains of Klebsiella isolated from urinary tract infections

Clinical and bacteriological findings in seven cases of urinary tract infection with cysteine-requiring strains of Klebsiella are described. The organisms were isolated from patients with long-standing urinary tract abnormalities and grew as small (c. 1 mm) colonies on MacConkey agar. The organisms failed to grow in a minimal medium supplemented with sodium sulphate but grew when the medium was supplemented with cysteine sulphinic acid, sodium sulphide or L-cysteine. The smallest amount of cysteine required for optimal growth in a chemically defined medium was 20 mg/L. Cysteine-requiring strains of Escherichia coli had previously been shown to require a similar amount of cysteine and to be unable to reduce sulphate to sulphite; this suggests a common influence in the selection of cysteine auxotrophs in vivo. However, the amino acid inhibited the growth of E. coli at concentrations which only slightly altered growth of the Klebsiella strains. Problems with the isolation, identification and sensitivity testing of cysteine-requiring Klebsiella were also observed and methods by which these may be minimised are suggested.     

Comparison of Bacteroides zoogleoformans strains isolated from soft tissue infections in cats with strains from periodontal disease in humans.

A total of 11 strains of Bacteroides zoogleoformans were isolated from 11 of 106 different cat subcutaneous "fight wound" abscesses and were among a total of 143 Bacteroides species isolated from these samples. They constituted 3.4% (11 of 325) of all anaerobic isolates. The cat strains and strains of B. zoogleoformans isolated from humans with periodontal disease were similar phenotypically as determined by biochemical reactions, polyacrylamide gel electrophoresis patterns of soluble proteins, and guanine plus cytosine ratios of DNA. Eight cat strains and five human strains tested had 45 to 54% DNA homology with the type strain of B. zoogleoformans. The eight cat strains and five human strains (excluding the type strain) were related by DNA homology at 70 to 77%. There was 85 to 90% intragroup DNA homology among the cat strains and 86 to 89% intragroup homology among the five human strains. The implications for epidemiology and human and animal ecology are discussed.     

New epidemiologic markers of Klebsiella oxytoca strains isolated from nosocomial infections

In addition to capsular antigen typing and biotyping (sorbose, d-tartrate, dulcitol), the epidemiology of Klebsiella oxytoca can be improved by new biochemical characterization. The authors propose to determine tetrathionate-reductase activity, D-melezitose fermentation and brown pigment production onto a mineral gluconate-ferric citrate medium. In order to assess the value of these new markers, several hospital epidemiological surveys were performed in France and Spain (112 strains studied).     

Characterization of Staphylococcus aureus strains isolated from humans in Argentina

Humans are a natural reservoir of Staphylococcus aureus and asymptomatic colonization is far more common than infection. The aim of this work was to characterize genotypically 68 S. aureus strains isolated from nasal swabs of healthy people and from human clinical infections. A total of fourteen (20%) strains were susceptible to all the antimicrobials tested. The strains isolated from nasal swabs showed the lowest percentages of resistance. Resistance to one or more than one antibiotics tested was detected in 83% and 70% of the S. aureus strains isolated from clinical infections and nasal swabs, respectively. All of the 68 S. aureus strains were subject to RAPD-PCR analysis. Cluster A-I grouped 42 (87%) clinical infection strains and cluster A-II grouped 13 (65%) strains isolated from nasal swabs suggesting a genetic relationship among S. aureus strains. Cluster A-II grouped 65% of the S. aureus strains associated with the anterior nares, suggesting that these strains may be adapted to this site. Furthermore, five RAPD profiles isolated from nasal swabs, belonged to clusters B to F, were similar to strains isolated from clinical infection, suggesting that they might have a high propensity to cause disease. The results of the present study allow a characterization of S. aureus strains isolated from humans and shows that some S. aureus genotypes from nasal swabs are similar to the genotypes obtained from clinical infections, suggesting that clinical isolates may be originated from human normal flora.     

Epidemiology and typing of Staphylococcus aureus strains isolated from bloodstream infections

We carried out an epidemiological study covering 2,365,067 patient days of hospitalization between 2000 and 2003. During this time, 413 Staphylococcus aureus bloodstream infections occurred. This corresponds to 15% of the 2,676 bloodstream infections observed during this period in the 31 hospitals in our region of France, which has 2.5 million inhabitants. The incidence of nosocomial S. aureus bloodstream infections was 0.11 per 1,000 days of hospitalization. The prevalence of methicillin-resistant S. aureus (MRSA) strains, of which 13% were nonmultiresistant MRSA (NORSA), was 33%, and this percentage was stable over the 4 years. In contrast, the prevalence of S. aureus strains susceptible to methicillin but resistant to quinolones or susceptible to methicillin but multiresistant to antibiotics (EMSSA strains) increased from 4% in 2000 to 23% in 2003. As previously reported, MRSA strains were mostly recovered from nosocomial bloodstream infections, whereas NORSA strains-generally considered to be responsible for community-acquired infections-were always isolated from nosocomial bloodstream infections. Pulsed-field gel electrophoresis (PFGE) analysis of 109 MRSA strains and 15 EMSSA strains demonstrated clonal diffusion of the three major French MRSA clones and revealed considerable genetic heterogeneity among EMSSA strains. Although no epidemiologically related NORSA strains clustered in particular PFGE groups, the distribution of MRSA strains isolated from bloodstream infections according to the portal of entry (vascular devices, pulmonary, and urinary) was not random for the major PFGE clones, suggesting that each MRSA lineage displays particular virulence features.     

Staphylococcus aureus strains isolated from bloodstream infections changed significantly in 2006

We studied 358 Staphylococcus aureus strains isolated from bloodstream infections (BSI) observed during an epidemiological study covering 2,007,681 days of hospitalization in 32 healthcare institutions (HCIs) between 2004 and 2006. The strains were tested for antibiotic susceptibility and characterized genetically. The incidence of S. aureus BSI declined regularly through 2004 and 2005 and then significantly increased in 2006 (+80%). This was largely due to an increase in BSI involving methicillin-sensitive S. aureus (MSSA) strains and nonmultiresistant methicillin-resistant S. aureus (NORSA) strains. Ninety-six percent of the NORSA strains were resistant only to methicillin and fluoroquinolones. Most of the MSSA strains belonged to a small number of pulsed-field gel electrophoresis (PFGE) divisions and were associated with epidemic phenomena in HCIs. The NORSA strains also clustered into a limited number of PFGE divisions but could not be related to any local outbreak in HCIs. In 2006, there was a significant increase in the incidence of BSI associated with tst gene-positive MSSA strains (+275%) and the first three BSI associated with tst gene-positive MRSA were observed. PFGE data revealed a limited heterogeneity among the tst gene-positive strains without any outbreak in the HCIs. Our study underlines the need for infection control teams to focus efforts on preventing both MRSA and MSSA BSI. As recently demonstrated in vitro, fluoroquinolones may enhance horizontal transfer of virulence and antibiotic resistance genes. These antibiotics are widely used in France, so our findings raise the issue of whether their use has contributed to the acquisition of mecA and tst genes by S. aureus strains.     

Characterization of Rhodococcus equi-like bacterium isolated from a wound infection in a noncompromised host.

A case of superficial wound infection in a noncompromised host due to a Rhodococcus equi-like organism, acquired from soil, is described. The strain grew poorly at 37 degrees C and showed several differences from wild-type R. equi in biochemical tests and in its fatty acid and mycolic acid compositions.     

Isolation of Aeromonas hydrophila in diarrhea. Characterization of enterotoxinogenic strains and clinical relations

Aeromonas hydrophila is isolated from diarrhoea specimens with increasing frequency. The interest in this organism at the present time is related to the fact that it can produce a number of toxins, in particular alpha and beta cytotoxic haemolysins, an enterotoxin and various enzymes. The authors determined the frequency of isolation of this organism and tested the haemolytic, cytotoxic and enterotoxic effects of culture filtrates in all of the stool specimens received in their laboratory over a period of 9 months. At the same time, the clinical context was defined in order to demonstrate a relation between the aptitude of the strains to produce toxins and the presence of diarrhoea. The frequency of isolation of A. hydrophila was 0.88 per cent, which corresponds to 67 strains. 38 strains presented a haemolytic and/or enterotoxic activity, i.e. 57 per cent of the strains isolated. In diarrhoeal stools, 67 per cent of the A. hydrophila isolated produced at least one of the toxins, while in the group of patients without diarrhoea, only 38 per cent of the strains isolated produced toxins. The results obtained reveal a statistically significant correlation between the production of cytotoxic haemolysin and the presence of diarrhoea. In contrast, there was no correlation between the production of enterotoxin and the presence of diarrhoea. Twenty of the 67 strains ware isolated from children under the age of 2 years. In 40 per cent of cases, no other aetiology could be found for the diarrhoea, apart from the isolation of A. hydrophila.(ABSTRACT TRUNCATED AT 250 WORDS)