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PubMed1996

Relationships among pathogenic and nonpathogenic isolates of Fusarium oxysporum based on the partial sequence of the intergenic spacer region of the ribosomal DNA.

Using PCR, we amplified and sequenced approximately 1,000 bp of the 5' end of the intergenic spacer (IGS) of the rDNA in 15 isolates of Fusarium oxysporum and one isolate of F. subglutinans. Isolates were selected to represent diversity in our collection based on differences in pathogenic race, vegetative compatibility group (VCG), mitochondrial DNA (mtDNA) haplotype, IGS haplotype, and DNA fingerprint. The objective of this research was to clarify the origin of virulence within F. oxysporum, the relationship between pathogenic and nonpathogenic strains, and the evolution of the different races of F. oxysporum f. sp. melonis. Bootstrapped parsimony analysis of the partial IGS sequence data identified a phylogenetic tree with highly significant branches. The two F. oxysporum f. sp. melonis VCGs, 0131 and 0134, were separated into distinct lineages. Race was not distinguished by significant IGS sequence differences within the pathogen VCGs. One exception was a race 1 isolate which was associated with VCG 0131 but, based on both mtDNA and IGS haplotype, had greater affinity with VCG 0134. Two IGS sequence types were found in this race 1 isolate, one suggesting an affiliation with VCG 0131 and the other similar to isolates in VCG 0134. This may have resulted from past somatic or sexual interactions between F. oxysporum f. sp. melonis, VCGs 0131 and 0134. Nonpathogens that were vegetatively compatible with the pathogen were not closely related to the pathogen based on IGS sequence data. Thus, nonpathogens and pathogens may share common alleles at vegetative compatibility loci by coincidence rather than because of recent clonal derivation from a common ancestor.

Base Sequence↗

PubMed2003

Characterization of Pasteurella multocida associated with pneumonia in bighorn sheep.

Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Although P. multocida is not ordinarily associated with disease in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), numerous isolates were cultured in high numbers from free-ranging bighorn sheep in the Hells Canyon area of Idaho, Washington, and Oregon (USA) during the winter of 1995-96. Animals captured in Hells Canyon and held in captivity, and their offspring, also harbored P. multocida. Biochemical utilization tests on 90 isolates identified three subspecies: P. multocida multocida a (n = 54); P. multocida multocida b (n = 13); and P. multocida gallicida (n = 15); and a non-speciated biotype, U6 (n = 8). Genomic DNA digestion with restriction endonuclease Hha I separated the isolates into 62 unique restriction fragment length polymorphism profiles. Capsular type A was predominant (72% of isolates). Only one isolate type, which may have been transmitted from a feral goat, was capsular type D, possessed the structural gene, toxA, for dermonecrotoxin detected by polymerase chain reaction, and produced toxin as determined by monoclonal antibody immunoblot. In conclusion, bighorn sheep in this study carried diverse types of generally non-toxigenic P. multocida associated with epizootic pneumonia.

Animals↗

PubMed2002

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

Gene Order↗

PubMed2003

High genetic diversity of the immunodominant region of the feline calicivirus capsid gene in endemically infected cat colonies.

Feline calicivirus (FCV) is an important pathogen of domestic cats. In this study, we have determined the genetic diversity of FCV within four geographically separate colonies of endemically infected cats by sequencing the immunodominant and variable region E of the capsid gene. Comparison of isolates between colonies and between unrelated published sequences gave nucleotide distance values of 26-35% and 22-40%, respectively and suggested each colony was infected with a distinct virus strain. Comparison of isolates within individual endemically infected colonies showed nucleotide distance variability of 0-16%. This was greater than distances previously reported for epidemiologically related isolates from cases of acute disease (0-5%) and was consistent with the evolution of FCV from a single distinct ancestor sequence in each colony. The pattern of nucleotide substitutions generating the observed intra-colony diversity was associated with strong evidence for positive selection acting on immunodominant regions of the FCV capsid protein. We suggest that endemically infected colonies of cats may be important generators of genetic diversity for FCV and that this may ultimately lead to the generation of new strains.

Amino Acid Sequence↗

PubMed1983

Bacterial plasmids--an overview.

Resistance to antimicrobial substances and various factors contributing to pathogenicity are only some of the bacterial traits that can be determined by plasmid genes. Not all plasmids are involved with resistance, nor is all resistance to antimicrobials plasmid-mediated. Nevertheless, the impact of R-plasmids on the choice of antimicrobial therapy is substantial, especially in hospital and when dealing with certain community pathogens, such as Haemophilus influenzae. The discovery of transposition elements has helped us to understand the rapidity with which certain resistance determinants develop and become disseminated among diverse microbial species. Similarly, not all determinants of microbial pathogenicity are plasmid encoded and the genes for many toxins, cellular attachment pili, iron sequestration systems, etc. are found on the bacterial chromosomes. In Yersinia, pathogenicity is even more complex in that plasmid determinants play an essential role, but only in concert with chromosomal genes. The particularly significant features of such pathogenicity-determined plasmid-mediated genes is not that they are found on extrachromosomal elements, but that they may possess greater genetic mobility. Another consequence of the presence of virulence genes on bacterial plasmids is that it makes them easier to study by the newer techniques of microbial genetics and molecular biology.

Anti-Bacterial Agents↗

PubMed1995

Genetic diversity of Burkholderia solanacearum (synonym Pseudomonas solanacearum) race 3 in Kenya.

Genetic diversity among isolates of the bacterial plant pathogen Burkholderia solanacearum (synonym Pseudomonas solanacearum) race 3 biovar II of Kenya was determined by PCR with repetitive sequences (ERIC and BOX repetitive primer sets) and pulsed-field gel electrophoresis of genomic DNA digested by rare-cutting restriction endonucleases (RC-PFGE). The study comprised 46 isolates collected during 1992 from the major potato-growing regions of Kenya (45 were identified as race 3 biovar II, and 1 belonged to race 3 biovar N2) and 39 reference isolates from 19 other countries. RC-PFGE identified 10 distinct profile types among the Kenyan race 3 biovar II isolates (29 of the isolates exhibited identical profiles) and a further 27 distinct profile types among the reference isolates. ERIC and BOX primer sets were unable to differentiate race 3 biovar II isolates within the Kenyan population but differentiated a further two distinct profile types among the reference isolates. The race 3 biovar N2 isolate had a highly distinct RC-PFGE and repetitive sequence PCR profile. Statistical analysis of the data identified biogeographic trends consistent with conclusions drawn from previous studies on the origin and worldwide dissemination of race 3 biovar II isolates; however, genomic fingerprinting by RC-PFGE revealed a level of genetic diversity previously unrealized.

Genetic Variation↗

PubMed2005

Effects of microbial community diversity on the survival of Pseudomonas aeruginosa in the wheat rhizosphere.

Ecological theory suggests that microbial communities with greater microbial diversity would be less susceptible to invasion by potential opportunistic pathogens. We investigated whether the survival of the opportunistic pathogen Pseudomonas aeruginosa in the wheat rhizosphere would be affected by the presence of natural and constructed microbial communities of various diversity levels. Three levels of microbial community diversity were derived from wheat roots by a dilution/extinction approach. These wheat rhizosphere inocula, as well as a gnotobiotic microbial community consisting of seven culturable wheat rhizobacterial isolates, were introduced into the nutrient solution of hydroponically grown wheat plants on the day of planting. Phenotypic characterization of the culturable microbial communities on R2A medium, Shannon microbial diversity index, community-level physiological profiles, and terminal restriction fragment length polymorphisms were used to assess the varying microbial diversity levels. At day 7 the roots were invaded with P. aeruginosa and the number of P. aeruginosa colony forming units per root were measured at day 14. The average number of surviving P. aeruginosa cells was 3.52, 4.90, 7.18, 6.65 log(10) cfu/root in the high, medium, low, and gnotobiotic microbial community diversity level treatments, respectively. The invasibility of the rhizosphere communities by P. aeruginosa was inversely related to the level of diversity from the dilution extinction gradient. The gnotobiotic community did not confer protection against P. aeruginosa invasion. Although these data indicate that invasibility is inversely related to diversity, further study is needed to both reproduce these findings and define the specific mechanisms of the diversity effect.

Biodiversity↗

PubMed1993

A small diffusible signal molecule is responsible for the global control of virulence and exoenzyme production in the plant pathogen Erwinia carotovora.

Virulence of the plant pathogen Erwinia carotovora subsp. carotovora is dependent on the production and secretion of a complex arsenal of plant cell wall-degrading enzymes. Production of these exoenzymes is controlled by a global regulatory mechanism. A virulent mutants in one of the regulatory loci, expI, show a pleiotropic defect in the growth phase-dependent transcriptional activation of exoenzyme gene expression. The expI gene encodes a 26 kDa polypeptide that is structurally and functionally related to the luxI gene product of Vibrio fischeri. Functional similarity of expI and luxI has been demonstrated by reciprocal genetic complementation experiments. LuxI controls bioluminescence in V.fischeri in a growth phase-dependent manner by directing the synthesis of the diffusible autoinducer, N-(3-oxohexanoyl) homoserine lactone. E.c. subsp. carotovora expI+ strains or Escherichia coli harboring the cloned expI gene excrete a small diffusible signal molecule that complements the expI mutation of Erwinia as well as a luxI mutation of V.fischeri. This extracellular complementation can also be achieved by E.coli harboring the luxI gene from V.fischeri or by adding the synthetic V.fischeri autoinducer. Both the production of the plant tissue-macerating exoenzymes and the ability of the bacteria to propagate in planta are restored in expI mutants by autoinducer addition. These data suggest that the same signal molecule is employed in control of such diverse processes as virulence in a plant pathogen and bioluminescence in a marine bacterium, and may represent a general mechanism by which bacteria modulate gene expression in response to changing environmental conditions.

Amino Acid Sequence↗

PubMed1994

Clonotypic dominance and variable gene elements of pathogenic anti-DNA autoantibodies from a single patient with lupus.

This study explores the usage and diversity of the variable gene elements expressed by human lupus antibodies to DNA bearing the 0-81 idiotype, a marker of pathogenic anti-DNA autoantibodies. Rather than studying DNA-specific clonotypes from different patients, a panel of idiotype positive anti-DNA autoantibody-secreting clones from a single individual were analysed. By cloning and nucleotide-sequencing the heavy-chain variable gene segments, evidence was found for dominance of clonotypic patterns. Also noted was a high rate of diversification among the variable (VH), diversity (DH) and junctional (JH) gene segments utilized, with a pattern of mutations indicative of antigenic selection. These features suggest that the clones secreting the lupus pathogenic autoantibodies have been selected over multiple generations through an affinity-maturation process that is reminiscent of antigen-driven immune responses.

Adult↗

PubMed1998

Higher-order predators and the regulation of insect herbivore populations.

Empirical research has not supported the prediction that populations of terrestrial herbivorous arthropods are regulated solely by their natural enemies. Instead, both natural enemies (top-down effects) and resources (bottom-up effects) may play important regulatory roles. This review evaluates the hypothesis that higher-order predators may constrain the top-down control of herbivore populations. Natural enemies of herbivorous arthropods generally are not top predators within terrestrial food webs. Insect pathogens and entomopathogenic nematodes inhabiting the soil may be attacked by diverse micro- and mesofauna. Predatory and parasitic insects are attacked by their own suite of predators, parasitoids, and pathogens. The view of natural enemy ecology that has emerged from laboratory studies, where natural enemies are often isolated from all elements of the biotic community except for their hosts or prey, may be an unreliable guide to field dynamics. Experimental work suggests that interactions of biological control agents with their own natural enemies can disrupt the effective control of herbivore populations. Disruption has been observed experimentally in interactions of bacteria with bacteriophages, nematodes with nematophagous fungi, parasitoids with predators, parasitoids with hyperparasitoids, and predators with other predators. Higher-order predators have been little studied; manipulative field experiments will be especially valuable in furthering our understanding of their roles in arthropod communities.

Animals↗

PubMed1994

The role of infectious disease, inbreeding and mating preferences in maintaining MHC genetic diversity: an experimental test.

In house mice, and probably most mammals, major histocompatibility complex (MHC) gene products influence both immune recognition and individual odours in an allele-specific fashion. Although it is generally assumed that some form of pathogen-driven balancing selection is responsible for the unprecedented genetic diversity of MHC genes, the MHC-based mating preferences observed in house mice are sufficient to account for the genetic diversity of MHC genes found in this and other vertebrates. These MHC disassortative mating preferences are completely consistent with the conventional view that pathogen-driven MHC heterozygote advantage operates on MHC genes. This is because such matings preferentially produce MHC-heterozygours progeny, which could enjoy enhanced disease resistance. However, such matings could also function to avoid genome-wide inbreeding. To discriminate between these two hypotheses we measured the fitness consequences of both experimentally manipulated levels of inbreeding and MHC homozygosity and heterozygosity in semi-natural populations of wild-derived house mice. We were able to measure a fitness decline associated with inbreeding, but were unable to detect fitness declines associated with MHC homozygosity. These data suggest that inbreeding avoidance may be the most important function of MHC-based mating preferences and therefore the fundamental selective force diversifying MHC genes in species with such mating patterns. Although controversial, this conclusion is consistent with the majority of the data from the inbreeding and immunological literature.

Animals↗

PubMed2003

The role of acquired immunity and periodontal disease progression.

Our understanding of the pathogenesis in human periodontal diseases is limited by the lack of specific and sensitive tools or models to study the complex microbial challenges and their interactions with the host's immune system. Recent advances in cellular and molecular biology research have demonstrated the importance of the acquired immune system not only in fighting the virulent periodontal pathogens but also in protecting the host from developing further devastating conditions in periodontal infections. The use of genetic knockout and immunodeficient mouse strains has shown that the acquired immune response-in particular, CD4+ T-cells-plays a pivotal role in controlling the ongoing infection, the immune/inflammatory responses, and the subsequent host's tissue destruction. In particular, studies of the pathogen-specific CD4+ T-cell-mediated immunity have clarified the roles of: (i) the relative diverse immune repertoire involved in periodontal pathogenesis, (ii) the contribution of pathogen-associated Th1-Th2 cytokine expressions in periodontal disease progression, and (iii) micro-organism-triggered periodontal CD4+ T-cell-mediated osteoclastogenic factor, 'RANK-L', which is linked to the induction of alveolar bone destruction in situ. The present review will focus on some recent advances in the acquired immune responses involving B-cells, CD8+ T-cells, and CD4+ T-cells in the context of periodontal disease progression. New approaches will further facilitate our understanding of their underlying molecular mechanisms that may lead to the development of new treatment modalities for periodontal diseases and their associated complications.

Aggregatibacter actinomycetemcomitans↗

PubMed1984

Ecology of pathogenic yeasts in Amazonian soil.

In an investigation of Amazonian soil as a natural reservoir for pathogenic fungi, 1,949 soil samples collected from diverse geographical and ecological settings of the Brazilian Amazon Basin were analyzed for the presence of non-keratinophilic fungi by the indirect mouse inoculation procedure and for the presence of keratinophilic fungi by the hair bait technique. All soil samples were acidic with low pH values. From 12% of the soil samples, 241 yeast and yeastlike isolates pertaining to six genera and 82 species were recovered, of which 63% were Torulopsis and 26% were Candida species. Nine fungi with known pathogenic potentials were encountered among 43% (104) of the isolates: T. glabrata, C. guilliermondii, C. albicans, C. pseudotropicalis, C. stellatoidea, C. tropicalis, Rhodotorula rubra, and Wangiella dermatitidis. The yeast flora was marked by species diversity, low frequency of each species, random geographical distribution, and an apparent lack of species clustering. The composition and distribution of the yeast flora in soil differed from those of the yeast flora harbored by bats, suggesting that the Amazonian external environment and internal bat organs act as independent natural habitats for yeasts.

Animals↗

PubMed2006

Microbial ecology of drinking water distribution systems.

The supply of clean drinking water is a major, and relatively recent, public health milestone. Control of microbial growth in drinking water distribution systems, often achieved through the addition of disinfectants, is essential to limiting waterborne illness, particularly in immunocompromised subpopulations. Recent inquiries into the microbial ecology of distribution systems have found that pathogen resistance to chlorination is affected by microbial community diversity and interspecies relationships. Research indicates that multispecies biofilms are generally more resistant to disinfection than single-species biofilms. Other recent findings are the increased survival of the bacterial pathogen Legionella pneumophila when present inside its protozoan host Hartmannella vermiformis and the depletion of chloramine disinfectant residuals by nitrifying bacteria, leading to increased overall microbial growth. Interactions such as these are unaccounted for in current disinfection models. An understanding of the microbial ecology of distribution systems is necessary to design innovative and effective control strategies that will ensure safe and high-quality drinking water.

Animals↗

PubMed2002

Mycobacterium haemophilum infections in heart transplant recipients: case report and review of the literature.

Non-tuberculous mycobacteria are becoming increasingly important pathogens among transplant recipients. We report a case of disseminated Mycobacterium haemophilum infection in a heart transplant recipient, manifesting as cellulitis, subcutaneous nodules, septic arthritis, and pneumonitis. Our case illustrates diverse challenges in the identification and treatment of this pathogen, such as its unique culture requirements and variable antimicrobial susceptibilities. Heightened clinical suspicion is necessary to establish a timely diagnosis so that optimal treatment can be administered.

Female↗

PubMed2003

'Order from disorder sprung': recognition and regulation in the immune system.

Milton's epic poem Paradise lost supplies a colourful metaphor for the immune system and its responses to pathogens. With the role of Satan played by pathogens seeking to destroy the paradise of human health, GOD intervenes and imposes order out of chaos. In this context, GOD means 'generation of diversity': the capacity of the innate and specific immune responses to recognize and eliminate a universe of pathogens. Thus, the immune system can be thought of as an entity that self-assembles the elements required to combat bodily invasion and injury. In so doing, it brings to bear the power of specific recognition: the ability to distinguish self from non-self, and the threatening from the benign. This ability to define and protect self is evolutionarily very old. Self-recognition and biochemical and barrier defences can be detected in primitive organisms, and elements of these mechanisms are built upon in an orderly way to establish the mammalian immune system. Innate immune responses depend on the use of a limited number of germline-encoded receptors to recognize conserved molecular patterns that occur on the surfaces of a broad range of pathogens. The B and T lymphocytes of the specific immune response use complex gene-rearrangement machinery to generate a diversity of antigen receptors capable of recognizing any pathogen in the universe. Binding to receptors on both innate and specific immune-system cells triggers intricate intracellular signalling pathways that lead to new gene transcription and effector-cell activation. And yet, regulation is imposed on these responses so that Paradise is not lost to the turning of the immune system onto self-tissues, the spectre of autoimmunity. Lymphocyte activation requires multiple signals and intercellular interactions. Mechanisms exist to establish tolerance to self by the selection and elimination of cells recognizing self-antigens. Immune system cell populations are reduced by programmed cell death once the pathogen threat is resolved. Once Paradise has been regained, memory cells remain in the body to sharply reduce the impact of a second exposure to a pathogen. Vaccination programs take advantage of this capacity of the human immune system for immunological memory, sparing millions the suffering associated with disease scourges. Thus does the order of the immune response spring from the disorder of pathogen attacks, and thus is Paradise preserved.

Adaptation, Physiological↗

PubMed2004

Intraspecific diversity within Diaporthe helianthi: evidence from rDNA intergenic spacer (IGS) sequence analysis.

Diaporthe helianthi is the causal agent of sunflower stem canker, a serious pathogen of sunflower in Europe but recorded sporadically in Italy. The genetic diversity of D. helianthi isolates from different geographic origins (Argentina, France, Italy, Yugoslavia, Romania) was investigated using IGS sequences. A 400 bp fragment of the portion of the IGS region flanking the 5' end of the 18S gene was amplified from each isolate. The aligned nucleotide sequences showed intraspecific sequence homology from 99-100% among French/Yugoslavian isolates to 95-100% among Italian isolates. French/Yugoslavian isolates shared 90-92% sequence homology with Italian isolates. The phylogenetic tree obtained from the aligned data revealed three separate groups. Group 1 included all isolates from France and former Yugoslavia and one isolate from Argentina; Group 2 included all Italian isolates and one isolate from Argentina. The most distantly related isolate was that from Romania (Group 3). The average genetic distances among isolates within Group 1 and within Group 2 were 0.22 and 3.29 respectively. The analysis showed that all isolates originating from countries where severe outbreaks of the disease are reported annually (France and former Yugoslavia) form a well defined taxon characterized by relatively low variability. This group is distinct from the group formed by isolates originating from Italy, whose variability is relatively much higher. Results obtained revealed a marked differentiation among pathogen isolates, and members of Group 1 seem not yet to have spread into Italian sunflower-growing areas.

Ascomycota↗

PubMed1999

Suppression of gene silencing: a general strategy used by diverse DNA and RNA viruses of plants.

In transgenic and nontransgenic plants, viruses are both initiators and targets of a defense mechanism that is similar to posttranscriptional gene silencing (PTGS). Recently, it was found that potyviruses and cucumoviruses encode pathogenicity determinants that suppress this defense mechanism. Here, we test diverse virus types for the ability to suppress PTGS. Nicotiana benthamiana exhibiting PTGS of a green fluorescent protein transgene were infected with a range of unrelated viruses and various potato virus X vectors producing viral pathogenicity factors. Upon infection, suppression of PTGS was assessed in planta through reactivation of green fluorescence and confirmed by molecular analysis. These experiments led to the identification of three suppressors of PTGS and showed that suppression of PTGS is widely used as a counter-defense strategy by DNA and RNA viruses. However, the spatial pattern and degree of suppression varied extensively between viruses. At one extreme, there are viruses that suppress in all tissues of all infected leaves, whereas others are able to suppress only in the veins of new emerging leaves. This variation existed even between closely related members of the potexvirus group. Collectively, these results suggest that virus-encoded suppressors of gene silencing have distinct modes of action, are targeted against distinct components of the host gene-silencing machinery, and that there is dynamic evolution of the host and viral components associated with the gene-silencing mechanism.

DNA Viruses↗