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PubMed1989

Hemagglutination and intestinal adherence properties of clinical and environmental isolates of non-O1 Vibrio cholerae.

Hemagglutination and intestinal adherence properties of non-O1 Vibrio cholerae were studied in vitro. No definite correlation between the cell-associated hemagglutinin titers and the intestinal adhesion indices was noted. Sugar- and glycoprotein-mediated inhibition data also indicated differences between the hemagglutination and adherence processes in respect to the receptor structures. Intestinal adherence of most V. cholerae strains could be inhibited to various extents by N-acetyl D-glucosamine. This observation provides a likely explanation for the ecological behavior of these organisms, which are known to associate themselves with chitinous (chitin:homopolymer of N-acetyl D-glucosamine) surfaces of zooplankton. The absence of any significant difference between the intestinal adherence indices of clinical and environmental isolates suggests that intestinal adhesion may be an essential but not sufficient prerequisite for colonization by and subsequent expression of pathogenicity of these microorganisms.

Animals↗

PubMed1995

Electron microscopic examination of factors influencing the expression of filamentous surface structures on clinical and environmental isolates of Aeromonas veronii Biotype sobria.

Strains of Aeromonas veronii biotype sobria isolated from clinical and environmental sources were examined for their expression of surface structures under a variety of culture conditions. When grown on solid media at 37 degrees C, more than 95% of bacteria from the majority of strains isolated from human diarrheal feces and chicken carcasses were non-piliated or expressed only a few pili of long, flexible morphology per cell. Strains isolated from water or other foods were much more likely to express pili. Heavily piliated strains (all sources) possessed pili of several morphological types, including long, flexible pili of varying widths and rigid pili of varying lengths. Expression of Pili was favored by growth at temperatures ca. 20 degrees C and below and growth in liquid medium. Most fecal strains expressed some pili under these conditions. In addition, other surface structures (fibrillar aggregates, fibrillar networks bundle-forming pili) were seen on some strains from most sources. These were also seen most frequently when bacteria were grown in liquid media at temperatures ca. 20 degrees C and below. Pili expression was not dramatically influenced by growth under anaerobic conditions, or in iron-depleted media, or by combinations of the above conditions. The role of the above surface structures in Aeromonas pathogenicity remains to be elucidated.

Aeromonas↗

PubMed2001

Molecular analysis of Vibrio cholerae O1, O139, non-O1, and non-O139 strains: clonal relationships between clinical and environmental isolates.

A total of 26 strains of Vibrio cholerae, including members of the O1, O139, and non-O1, non-O139 serogroups from both clinical and environmental sources, were examined for the presence of genes encoding cholera toxin (ctxA), zonula occludens toxin (zot), accessory cholera enterotoxin (ace), hemolysin (hlyA), NAG-specific heat-stable toxin (st), toxin-coregulated pilus (tcpA), and outer membrane protein (ompU), for genomic organization, and for the presence of the regulatory protein genes tcpI and toxR in order to determine relationships between epidemic serotypes and sources of isolation. While 22 of the 26 strains were hemolytic on 5% sheep blood nutrient agar, all strains were PCR positive for hlyA, the hemolysin gene. When multiplex PCR was used, all serogroup O1 and O139 strains were positive for tcpA, ompU, and tcpI. All O1 and O139 strains except one O1 strain and one O139 strain were positive for the ctxA, zot, and ace genes. Also, O1 strain VO3 was negative for the zot gene. All of the non-O1, non-O139 strains were negative for the ctxA, zot, ace, tcpA, and tcpI genes, and all of the non-O1, non-O139 strains except strain VO26 were negative for ompU. All of the strains except non-O1, non-O139 strain VO22 were PCR positive for the gene encoding the central regulatory protein, toxR. All V. cholerae strains were negative for the NAG-specific st gene. Of the nine non-ctx-producing strains of V. cholerae, only one, non-O1, non-O139 strain VO24, caused fluid accumulation in the rabbit ileal loop assay. The other eight strains, including an O1 strain, an O139 strain, and six non-O1, non-O139 strains, regardless of the source of isolation, caused fluid accumulation after two to five serial passages through the rabbit gut. Culture filtrates of all non-cholera-toxigenic strains grown in AKI media also caused fluid accumulation, suggesting that a new toxin was produced in AKI medium by these strains. Studies of clonality performed by using enterobacterial repetitive intergenic consensus sequence PCR, Box element PCR, amplified fragment length polymorphism (AFLP), and pulsed-field gel electrophoresis (PFGE) collectively indicated that the V. cholerae O1 and O139 strains had a clonal origin, whereas the non-O1, non-O139 strains belonged to different clones. The clinical isolates closely resembled environmental isolates in their genomic patterns. Overall, there was an excellent correlation among the results of the PCR, AFLP, and PFGE analyses, and individual strains derived from clinical and environmental sources produced similar fingerprint patterns. From the results of this study, we concluded that the non-cholera-toxin-producing strains of V. cholerae, whether of clinical or environmental origin, possess the ability to produce a new secretogenic toxin that is entirely different from the toxin produced by toxigenic V. cholerae O1 and O139 strains. We also concluded that the aquatic environment is a reservoir for V. cholerae O1, O139, non-O1, and non-O139 serogroup strains.

Animals↗

PubMed1996

In vitro susceptibilities of clinical and environmental isolates of Cryptococcus neoformans to five antifungal drugs.

A total of 53 Cryptococcus neoformans strains, including clinical and environmental Brazilian isolates, were tested for their susceptibilities to amphotericin B, 5-flucytosine, ketoconazole, fluconazole, and itraconazole. The tests were performed according to the National Committee of Clinical Laboratory Standards recommendations (document M27-P). In general, there was a remarkable homogeneity of results for all strains, and comparable MICs were found for environmental and clinical isolates. This paper represents the first contribution in which susceptibility data for Brazilian C. neoformans isolates are provided.

AIDS-Related Opportunistic Infections↗

PubMed1986

A survey of potential virulence factors in clinical and environmental isolates of Serratia marcescens.

One hundred and forty seven isolates of Serratia marcescens were collected from diverse clinical and environmental sources in south-east Texas. Natural isolates were compared with hospital strains for the occurrence of 12 potential virulence determinants. Their overall frequency was as follows: haemolytic activity 48%; lecithinase 95%; lipase 95%; motility 99%; pigmentation 24%; plasmid carriage 46%; proteolytic activity 98%; siderophore activity 99%; urease activity 5%; mannose-sensitive haemagglutination 96%; mannose-resistant haemagglutination 61%; and mannose-resistant type-K haemagglutination (MR/K-HA) 68%. Clinical strains demonstrated a significantly higher occurrence of MR/K-HA (p less than 0.001) and non-pigmentation (p less than 0.01) than environmental isolates.

Hemagglutination↗

PubMed1992

Enterotoxicity of clinical and environmental isolates of Aeromonas spp.

Of 147 isolates of three species of Aeromonas, 54 were from clinical and 93 from environmental sources. When tested for enterotoxin production, most of the isolates (56%) caused accumulation of fluid in rabbit ileal loops (RILs). Although large proportions of clinical and environmental isolates of A. caviae (55% and 65%, respectively) elicited such a response in RILs, isolates of A. hydrophila and A. sobria produced significantly more fluid (p less than 0.05). Furthermore, the environmental strains of A. hydrophila and A. sobria produced more fluid than the clinical isolates (p less than 0.05). The strains of Aeromonas spp. that caused little or no fluid accumulation in the initial experiments became enterotoxin producers after 1-3 passages through RILs, regardless of their source, and showed gradual enhancement of fluid outpouring after each passage. The present study suggests that all strains of these species of Aeromonas are potentially enterotoxigenic, whether from clinical or environmental sources.

Aeromonas↗

PubMed1983

Plasmid profiles of clinical and environmental isolates of Legionella pneumophila serogroup 1.

Clinical and environmental Legionella pneumophila serogroup 1 isolates from a single water source in Columbus, Ohio, exhibited five different plasmid profiles. The multiplicity of plasmid profiles observed within a single geographic area and a skewed distribution of isolates bearing these plasmid profiles within this area suggest that plasmid analyses will be useful in the study of the epidemiology of Legionnaires disease and the environmental distribution of legionellae.

Bacterial Infections↗

PubMed1997

Environmental isolation of Cryptococcus neoformans var. neoformans from Vellore.

We have isolated C. neoformans from various environmental niches, including our hospital environment. Twenty one samples of weathered pigeon droppings and 86 samples of Eucalyptus bark and debris were collected from various sites around the hospital and college. C. neoformans var. neoformans was isolated from 14 of the 21 (67%) samples of weathered droppings of pigeons collected from the ledges of windows of wards and departmental buildings in the hospital. C. neoformans was not isolated from any of the Eucalyptus samples. These findings document for the first time the presence of C. neoformans var. neoformans in the hospital environment. Although ubiquitous in the environment, its presence in hospital surroundings stresses the need for systematic cleaning of sites inhabited by pigeons. However, we failed to isolate C. neoformans from the limited number of Eucalyptus samples examined.

Animals↗

PubMed2003

Environmental isolation of Balamuthia mandrillaris associated with a case of amebic encephalitis.

This report describes the first isolation of the ameba Balamuthia mandrillaris from an environmental soil sample associated with a fatal case of amebic encephalitis in a northern California child. Isolation of the ameba into culture from autopsied brain tissue confirmed the presence of Balamuthia: In trying to locate a possible source of infection, soil and water samples from the child's home and play areas were examined for the presence of Balamuthia: The environmental samples (plated onto nonnutrient agar with Escherichia coli as a food source) contained, in addition to the ameba, a variety of soil organisms, including other amebas, ciliates, fungi, and nematodes, as contaminants. Presumptive Balamuthia amebas were recognized only after cultures had been kept for several weeks, after they had burrowed into the agar. These were transferred through a succession of nonnutrient agar plates to eliminate fungal and other contaminants. In subsequent transfers, axenic Naegleria amebas and, later, tissue cultures (monkey kidney cells) served as the food source. Finally, the amebas were transferred to cell-free axenic medium. In vitro, the Balamuthia isolate is a slow-growing organism with a generation time of approximately 30 h and produces populations of approximately 2 x 10(5) amebas per ml. It was confirmed as Balamuthia by indirect immunofluorescence staining with rabbit anti-Balamuthia serum and human anti-Balamuthia antibody-containing serum from the amebic encephalitis patient. The environmental isolate is similar in its antimicrobial sensitivities and identical in its 16S ribosomal DNA sequences to the Balamuthia isolate from the deceased patient.

Amebiasis↗

PubMed2025

Mapping genetic and phenotypic diversity of Pseudomonas aeruginosa across clinical and environmental isolation sites.

Pseudomonas aeruginosa is a clinically significant opportunistic pathogen adept at thriving in both host-associated and environmental settings. To define the extent to which P. aeruginosa isolates specialize across niches and identify genotype-phenotype correlates, we performed whole genome sequencing and comprehensive phenotypic characterization of 125 P. aeruginosa isolates from diverse clinical and environmental sites, evaluating virulence-associated traits, including motility, cytotoxicity, biofilm formation, pyocyanin production, and antimicrobial susceptibility. We identify that genomic diversity does not correlate with isolation source or most virulence phenotypes. Instead, we find that the two major P. aeruginosa clades (Groups A and B) delineate phylogeny and cytotoxicity, with Group B strains showing significantly higher cytotoxicity than Group A. Sequence analysis revealed previously uncharacterized alleles of genes encoding type III secretion effector proteins. We observed high variability amongst strains and isolation sources in all four assayed virulence phenotypes. Antimicrobial resistance (AMR) is exclusively observed in clinical isolates, not environmental, reflecting antibiotic exposure-driven selection. Bacterial GWAS revealed a statistically significant association between cytotoxicity and exoU presence, and we identified a novel exoU allelic variant with decreased cytotoxicity, demonstrating that functional diversity within well-characterized virulence factors may still influence pathogenic outcomes. In summary, our analyses of 125 diverse isolates suggest that the ability of P. aeruginosa to thrive across diverse niches is driven by broadly conserved genetic repertoire rather than niche-specific accessory genes.

Journal Article↗

PubMed2000

Arsenic and cadmium resistance in environmental isolates of Yersinia enterocolitica and Yersinia intermedia.

Environmental strains of Yersinia enterocolitica representing biotype 1A lack virulence plasmid (pYV) and are regarded as non-pathogenic. Though these occupy a diverse range of environmental niches, nothing is known about their resistance to heavy metals. The minimal inhibitory concentrations (MICs) of various metal ions, namely Ag+, Cu2+, Zn2+, Cd2+, As5+, and As3+, for strains of Yersinia enterocolitica (biotype 1A) and Yersinia intermedia (biotypes 1, 2, and 4), isolated from sewage effluents or pork, were determined. All isolates were resistant (MICs 2.5-5 mM) to Cd2+. The MICs of arsenic varied with bacterial strain and the chemical species of the arsenic used. For the majority of the strains, however, it was between 5-10 mM of Na2HAsO4.7H2O and NaAsO2, and 0.625-2.5 mM of As2O3. Except for one isolate, MICs of Ag+, Cu2+, and Zn2+ for these strains were in the range of 0.3-0.625 mM.

Animals↗

PubMed1987

Isolated environmental tilt associated with lateral medullary compression by dolichoectasia of the vertebral artery. Is there a cause and effect relationship?

An elderly woman with the persistent isolated symptom of monocular and binocular tilting of environmental objects, was found on magnetic resonance imaging to have compression of the lateral medulla by an ectatic vertebral artery. The double Maddox rod test showed no relative torsion of either globe although 5 degrees of right excyclotorsion and left incyclotorsion (rightward ocular counter-rolling) was detected on testing with monocular viewing. There was no accompanying hyperdeviation, nystagmus or other neurologic signs. Symmetrical tilting of each globe associated with head tilt and skew deviation (the ocular tilt reflex) may be caused by lesions involving otolith connections. Isolated environmental tilt may herald the onset of lateral medullary infarction (Wallenberg's syndrome), or may be associated with vertebro-basilar insufficiency. Could compression of otolith connections in the medulla cause persistent ocular counter-rolling without accompanying neurologic signs?

Aged↗

PubMed2003

Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a ubiquitous environmental bacterium capable of causing a variety of life-threatening human infections. The genetic basis for preferential infection of certain immunocompromised patients or individuals with cystic fibrosis by P. aeruginosa is not understood. To establish whether variation in the genomic repertoire of P. aeruginosa strains can be associated with a particular type of infection, we used a whole-genome DNA microarray to determine the genome content of 18 strains isolated from the most common human infections and environmental sources. A remarkable conservation of genes including those encoding nearly all known virulence factors was observed. Phylogenetic analysis of strain-specific genes revealed no correlation between genome content and infection type. Clusters of strain-specific genes in the P. aeruginosa genome, termed variable segments, appear to be preferential sites for the integration of novel genetic material. A specialized cloning vector was developed for capture and analysis of these genomic segments. With this capture system a site associated with the strain-specific ExoU cytotoxin-encoding gene was interrogated and an 80-kb genomic island carrying exoU was identified. These studies demonstrate that P. aeruginosa strains possess a highly conserved genome that encodes genes important for survival in numerous environments and allows it to cause a variety of human infections. The acquisition of novel genetic material, such as the exoU genomic island, through horizontal gene transfer may enhance colonization and survival in different host environments.

Bacterial Proteins↗

PubMed2003

Antifungal susceptibility of clinical and environmental isolates of Cryptococcus neoformans to four antifungal drugs determined by two techniques.

A total of 64 Cryptococcus neoformans strains, including clinical and environmental Brazilian isolates var. neoformans and var. gattii, were tested for susceptibility to amphotericin B, 5-flucytosine, fluconazole and itraconazole. The tests were performed according to the recommendations of National Committee of Clinical Laboratory Standards and the method of macrodilution in liquid medium of Shadomy et al. [Manual de Microbiologia Clínica, 4th ed. Buenos Aires: Editorial Medica Panamericana, 1987: 1229-38]. For most drugs there was a significant difference between the readings taken at 24 and 48 h with both methods. When the minimum inhibitory concentrations obtained by the two techniques were compared, significant differences were observed for amphotericin B and fluconazole. Overall, differences in drug susceptibility with respect to the origin of the isolates or the variety of the fungus were not observed. As an exception, the gattii variety exhibited a high resistance rate to amphotericin B when the technique of Shadomy et al. was applied, a fact possibly related to the greater difficulty for treatment of the disease caused by this fungal variety.

Amphotericin B↗

PubMed2003

Genetic characterization of environmental isolates of the Cryptococcus neoformans species complex from Brazil.

The genetic affiliation of a large number of isolates of the Cryptococcus neoformans species complex from environmental sources in Brazil has been investigated using amplified fragment length polymorphism (AFLP). The strains of C. neoformans isolated from a single tree, as well as from neighbouring trees, showed high similarity values (> 95%) of their AFLP patterns, thus suggesting considerable genetic homogeneity. The majority of isolates of C. neoformans belonged to AFLP genotype 1, and had serotype A and mating type alpha (= C. neoformans var. grubii). Three isolates belonged to AFLP genotype 2, with serotype D and mating type alpha (= C. neoformans var. neoformans). One isolate, obtained from a building in Rio de Janeiro inhabited by pigeons, belonged to the AD hybrid AFLP genotype 3. All isolates from trees of C. neoformans var. gattii (= C. gattii) belonged to AFLP genotype 6, and their banding patterns showed relatively low genetic homogeneity with a similarity value of about 76%. Isolates of this genotype occupy an environmental niche in the Americas, and they may cause disease in non-AIDS and AIDS patients as well.

Air Microbiology↗

PubMed1984

Biotyping and virulence factors in clinical and environmental isolates of Aeromonas species.

Biochemical characteristics and virulence factors were compared in 147 Aeromonas spp. isolated from patients with diarrhea and in 94 strains isolated from metropolitan water supplies in the same area during the same period. Fermentation of arabinose occurred with 58.5% of the environmental strains and 15% of the clinical isolates; 39.4% of the strains from water and 6.8% of the fecal isolates fermented salicin. The frequency of esculin hydrolysis was the same in both groups. Ninety-one percent of clinical isolates and 70.2% of environmental strains were enterotoxigenic and, except for four clinical isolates, all of these strains also produced hemolysins. Hemagglutination that was inhibited by fucose and mannose but not by galactose was found in 67% of the water isolates and 10.2% of the clinical strains. Although the distribution of several characteristics differs in clinical and environmental strains, many of the strains found in water have properties identical with those of the clinical isolates. We suggest that such strains may be potential enteric pathogens.

Aeromonas↗

PubMed1994

Interactions with lectins and agglutination profiles of clinical, food, and environmental isolates of Listeria.

On the basis of preliminary trials with 14 collection strains of Listeria, five lectins (Canavalia ensiformis, concanavalin A; Griffonia simplicifolia lectin I; Helix pomatia agglutinin; Ricinus communis agglutinin; and Triticum vulgaris wheat germ agglutinin) were selected to set up a microtiter agglutination assay. The lectin agglutination profiles of 174 clinical, food, and environmental strains of Listeria monocytogenes, Listeria innocua, and Listeria seeligeri were investigated. Data on the standard determination of the antigenic structure were available for clinical strains; nonclinical isolates were assigned to serogroup 1 or 4 with commercial antisera. The listeria-lectin interaction was related to serological type rather than species; in particular, the strains assigned to serogroup 1 or belonging to serovars 1/2a, 1/2b, 1/2c, 3a, 3b, and 7 were never agglutinated by G. simplicifolia lectin I. The five-lectin set proved to be capable of detecting differences between serologically identical isolates of L. monocytogenes. Of the 150 isolates of this species, 144 were distributed over 15 different lectin agglutination profiles and 6 autoagglutinated, the overall typeability being 96%. However, the profiles encountered among L. monocytogenes isolates were not randomly distributed. With strains assigned to serogroup 1 or belonging to serovars 1/2a, 1/2b, 1/2c, and 3b, the clinical isolates fell into only two of the eight patterns recorded overall; with strains of serogroup 4 and serovar 4b, food and environmental isolates were distributed over eight of the nine patterns found in total, while clinical isolates were distributed over five patterns. In a comparative study of 15 epidemiologically relevant isolates of L. monocytogenes from five distinct outbreaks, strains with identical phage types and/or DNA fingerprints displayed identical lectin profiles. The heterogeneity of agglutination profiles may form the basis of a new approach to L. monocytogenes typing.

Agglutination↗

PubMed1999

Distribution of two hemolytic toxin genes in clinical and environmental isolates of Aeromonas spp.: correlation with virulence in a suckling mouse model.

Previous studies have shown that two hemolytic toxins, HlyA and AerA, contribute to the virulence of Aeromonas hydrophila. A survey was performed to gauge the distribution of hlyA and aerA genes in clinical and environmental Aeromonas isolates. For A. hydrophila, A. veronii biotype sobria and A caviae, 96%, 12% and 35% of strains, respectively, were hlyA positive, whereas, 78%, 97%, 41%, respectively, were aerA positive. All virulent A. hydrophila isolates were hlyA+ aerA+. This genotype was most common in A. hydrophila (75.4%) followed by A. caviae (29.4%) and A. veronii biotype sobria (9.6%). For A. hydrophila, a two-hemolytic toxin model of virulence provides the best prediction of virulence in an animal model.

Aeromonas↗