Persistence of a Staphylococcus aureus small-colony variant under antibiotic pressure in vivo

Staphylococcus aureus small-colony variants (SCVs) have been implicated in chronic and persistent infections. Bovine mastitis induced by S. aureus is an example of an infection difficult to eradicate by conventional antimicrobial therapies. In this study, the ability to colonize mouse mammary glands and persist under antibiotic treatment was assessed for S. aureus Newbould and an isogenic hemB mutant, which exhibited the classical SCV phenotype. The hemB mutant showed a markedly reduced capacity to colonize tissues. However, although the hemB mutant was as susceptible as S. aureus Newbould to cephapirin in vitro, it was over a 100 times more persistent than the parental strain in the mammary glands when 1 or 2 mg kg(-1) doses were administrated. These results suggest that, although the hemB mutant has a reduced ability to colonize mammary glands, the SCV phenotype may account for the persistence of S. aureus under antibiotic pressure in vivo.     

The effect of the Agrobacterium tumefaciens attR mutation on attachment and root colonization differs between legumes and other dicots

Infections of wound sites on dicot plants by Agrobacterium tumefaciens result in the formation of crown gall tumors. An early step in tumor formation is bacterial attachment to the plant cells. AttR mutants failed to attach to wound sites of both legumes and nonlegumes and were avirulent on both groups of plants. AttR mutants also failed to attach to the root epidermis and root hairs of nonlegumes and had a markedly reduced ability to colonize the roots of these plants. However, AttR mutants were able to attach to the root epidermis and root hairs of alfalfa, garden bean, and pea. The mutant showed little reduction in its ability to colonize these roots. Thus, A. tumefaciens appears to possess two systems for binding to plant cells. One system is AttR dependent and is required for virulence on all of the plants tested and for colonization of the roots of all of the plants tested except legumes. Attachment to root hairs through this system can be blocked by the acetylated capsular polysaccharide. The second system is AttR independent, is not inhibited by the acetylated capsular polysaccharide, and allows the bacteria to bind to the roots of legumes.     

The Effect of the Agrobacterium tumefaciens attR Mutation on Attachment and Root Colonization Differs between Legumes and Other Dicots

Infections of wound sites on dicot plants by Agrobacterium tumefaciens result in the formation of crown gall tumors. An early step in tumor formation is bacterial attachment to the plant cells. AttR mutants failed to attach to wound sites of both legumes and nonlegumes and were avirulent on both groups of plants. AttR mutants also failed to attach to the root epidermis and root hairs of nonlegumes and had a markedly reduced ability to colonize the roots of these plants. However, AttR mutants were able to attach to the root epidermis and root hairs of alfalfa, garden bean, and pea. The mutant showed little reduction in its ability to colonize these roots. Thus, A. tumefaciens appears to possess two systems for binding to plant cells. One system is AttR dependent and is required for virulence on all of the plants tested and for colonization of the roots of all of the plants tested except legumes. Attachment to root hairs through this system can be blocked by the acetylated capsular polysaccharide. The second system is AttR independent, is not inhibited by the acetylated capsular polysaccharide, and allows the bacteria to bind to the roots of legumes.     

The role of Tir, EspA, and NleB in the colonization of cattle by Shiga toxin producing Escherichia coli O26:H11

Shiga toxin producing Escherichia coli (STEC) O26:H11 is an enteric pathogen capable of causing severe hemorrhagic colitis that can lead to hemolytic uremic syndrome. This organism is able to colonize cattle and human intestinal epithelial cells by secreting effectors via a type III secretion system (T3SS). In this investigation, we examined the role of 2 effectors, Tir and NleB, and the structural translocator component EspA in the adherence of STEC to epithelial cells and in the colonization of cattle. Isogenic deletion mutants were constructed and using microscopy and flow cytometry compared to the wild-type strain in their ability to adhere to HEp-2 cells. A competitive assay was also used to measure the capacity of the mutants to colonize the intestinal tract of cattle, where both the mutant and the parental strains were introduced orally at the same time. Genomic DNA was extracted from enriched fecal samples collected at various time points, and quantitative real-time PCR was used to quantify bacteria. A significant reduction in fecal shedding was observed, and adherence to HEp-2 cells was decreased for the tir and espA mutants. Deletion of the nleB gene did not have a significant effect on the adherence of HEp-2 cells; however, in an in vivo model, it strongly reduced the ability of STEC O26:H11 to colonize the bovine intestinal tract.     

Chlorine-induced damage to surface adhesions during sublethal injury of enterotoxigenic Escherichia coli.

A comparison of the adhesive ability of noninjured and chlorine-injured enterotoxigenic Escherichia coli was made by in vitro attachment to human peripheral leukocytes. Chlorination selected for noninjured cells with greater capabilities for colonizing the small intestine. Injured populations exhibited reduced association with leukocytes. Maximum reduction was seen in populations with greater than 80% injury. These cells demonstrated less adhesive ability than nonpiliated populations. Electron micrographs suggested that reduced adhesive ability was due to the loss of surface structures as a consequence of sublethal chlorination. The data imply a reduced ability among chlorine-injured pathogens to colonize the small intestine and initiate disease.     

Metabolic diversity in Campylobacter jejuni enhances specific tissue colonization

Campylobacter jejuni is a leading cause of foodborne illness in industrialized countries. This pathogen exhibits significant strain-to-strain variability, which results in differences in virulence potential and clinical presentations. Here, we report that acquisition of the capacity to utilize specific nutrients enhanced the ability of a highly pathogenic strain of C. jejuni to colonize specific tissues. The acquisition of a gene encoding a gamma-glutamyltranspeptidase enabled this strain to utilize glutamine and glutathione and enhanced its ability to colonize the intestine. Furthermore, the acquisition of a DNA segment, which added a sec-dependent secretion signal to an otherwise cytoplasmic asparaginase, allowed this pathogen to utilize asparagine and to more efficiently colonize the liver. Our results reveal that subtle genetic changes in a bacterial pathogen result in significant changes in its ability to colonize specific tissues. In addition, these studies revealed remarkably specific nutritional requirements for a pathogen to effectively colonize different tissues.     

Control of virulence by the two-component system CiaR/H is mediated via HtrA, a major virulence factor of Streptococcus pneumoniae

The CiaR/H two-component system is involved in regulating virulence and competence in Streptococcus pneumoniae. The system is known to regulate many genes, including that for high-temperature requirement A (HtrA). This gene has been implicated in the ability of the pneumococcus to colonize the nasopharynx of infant rats. We reported previously that deletion of the gene for HtrA made the pneumococcal strains much less virulent in mouse models, less able to grow at higher temperatures, and more sensitive to oxidative stress. In this report, we show that the growth phenotype as well as sensitivity to oxidative stress of Delta ciaR mutant was very similar to that of a Delta htrA mutant and that the expression of the HtrA protein was reduced in a ciaR-null mutant. Both the in vitro phenotype and the reduced virulence of Delta ciaR mutant could be restored by increasing the expression of HtrA.     

An Ribonuclease T2 Family Protein Modulates Acinetobacter baumannii Abiotic Surface Colonization

Acinetobacter baumannii is an emerging bacterial pathogen of considerable medical concern. The organism''s transmission and ability to cause disease has been associated with its propensity to colonize and form biofilms on abiotic surfaces in health care settings. To better understand the genetic determinants that affect biomaterial attachment, we performed a transposon mutagenesis analysis of abiotic surface-colonization using A. baumannii strain 98-37-09. Disruption of an RNase T2 family gene was found to limit the organism''s ability to colonize polystyrene, polypropylene, glass, and stainless steel surfaces. DNA microarray analyses revealed that in comparison to wild type and complemented cells, the RNase T2 family mutant exhibited reduced expression of 29 genes, 15 of which are predicted to be associated with bacterial attachment and surface-associated motility. Motility assays confirmed that RNase T2 mutant displays a severe motility defect. Taken together, our results indicate that the RNase T2 family protein identified in this study is a positive regulator of A. baumannii''s ability to colonize inanimate surfaces and motility. Moreover, the enzyme may be an effective target for the intervention of biomaterial colonization, and consequently limit the organism''s transmission within the hospital setting.     

Complement resistance is essential for colonization of the digestive tract of Hirudo medicinalis by Aeromonas strains

From the crop of the medicinal leech, Hirudo medicinalis, only Aeromonas veronii bv. sobria can be cultured consistently. Serum-sensitive A. veronii mutants were unable to colonize H. medicinalis, indicating the importance of the mammalian complement system for this unusual simplicity. Complementation of one selected mutant restored its ability to colonize. Serum-sensitive mutants are the first mutant class with a colonization defect for this symbiosis.     

Studies on Prn variation in the mouse model and comparison with epidemiological data

The virulence factor pertactin (Prn) is a component of pertussis vaccines and one of the most polymorphic Bordetella pertussis antigens. After the introduction of vaccination shifts in predominant Prn types were observed and strains with the Prn vaccine type (Prn1) were replaced by strains carrying non-vaccine types (Prn2 and Prn3), suggesting vaccine-driven selection. The aim of this study was to elucidate the shifts observed in Prn variants. We show that, although Prn2 and Prn3 circulated in similar frequencies in the 1970s and 1980s, in the 1990s Prn2 strains expanded and Prn3 strains disappeared, suggesting that in vaccinated populations Prn2 strains are fitter than Prn3 strains. We established a role for Prn in the mouse model by showing that a Prn knock-out (Prn-ko) mutation reduced colonization in trachea and lungs. Restoration of the mutation resulted in a significant increase in colonization compared to the knock-out mutant. The ability of clinical isolates with different Prn variants to colonize the mouse lung was compared. Although these isolates were also polymorphic at other loci, only variation in the promoter for pertussis toxin (ptxP) and Prn were found to contribute significantly to differences in colonization. Analysis of a subset of strains with the same ptxP allele revealed that the ability to colonize mice decreased in the order Prn1>Prn2 and Prn3. Our results are consistent with the predominance of Prn1 strains in unvaccinated populations. Our results show that ability to colonize mice is practically the same for Prn2 and Prn3. Therefore other factors may have contributed to the predominance of Prn2 in vaccinated populations. The mouse model may be useful to assess and predict changes in the B. pertussis population due to vaccination.     

Development of a Root Colonization Bioassay for Rapid Screening of Rhizobacteria for Potential Biocontrol Agents

The ability to colonize roots is a sine qua non condition for a rhizobacteria to be considered a true plant growth‐promoting rhizobacteria (PGPR). A simple screening method to detect such a potential ability of PGPR is described. Tomato seeds were surface sterilized for 30 s in 50% ethanol and this was followed by 3 min dipping in 2% NaClO. They were then washed three times in sterile water, left immersed in a propagule suspension of the rhizobacteria for 24 h, and transferred onto sterile 0.6% water‐agar in tubes. The young, developing root system shows a tendency to grow downwards in the agar‐gel column. When the rhizobacterium has a potential ability to colonize roots it is possible to visualize, by transparency, bacterial growth (turbid, milky and narrow zone) along and around roots. Testing 500 rhizobacteria isolated from tomato rhizosphere for their ability to induce systemic resistance against Pseudomonas syringae pv. tomato, 28 of them did reduce infection to less than 40% and all 28 colonized roots according to the described bioassay. Therefore the bioassay may turn into an important auxiliary tool for helping in selecting rhizobacteria with PGPR potentiality.     

Succinoglycan Is Required for Initiation and Elongation of Infection Threads during Nodulation of Alfalfa by Rhizobium meliloti

Rhizobium meliloti Rm1021 must be able to synthesize succinoglycan in order to invade successfully the nodules which it elicits on alfalfa and to establish an effective nitrogen-fixing symbiosis. Using R. meliloti cells that express green fluorescent protein (GFP), we have examined the nature of the symbiotic deficiency of exo mutants that are defective or altered in succinoglycan production. Our observations indicate that an exoY mutant, which does not produce succinoglycan, is symbiotically defective because it cannot initiate the formation of infection threads. An exoZ mutant, which produces succinoglycan without the acetyl modification, forms nitrogen-fixing nodules on plants, but it exhibits a reduced efficiency in the initiation and elongation of infection threads. An exoH mutant, which produces symbiotically nonfunctional high-molecular-weight succinoglycan that lacks the succinyl modification, cannot form extended infection threads. Infection threads initiate at a reduced rate and then abort before they reach the base of the root hairs. Overproduction of succinoglycan by the exoS96::Tn5 mutant does not reduce the efficiency of infection thread initiation and elongation, but it does significantly reduce the ability of this mutant to colonize the curled root hairs, which is the first step of the invasion process. The exoR95::Tn5 mutant, which overproduces succinoglycan to an even greater extent than the exoS96::Tn5 mutant, has completely lost its ability to colonize the curled root hairs. These new observations lead us to propose that succinoglycan is required for both the initiation and elongation of infection threads during nodule invasion and that excess production of succinoglycan interferes with the ability of the rhizobia to colonize curled root hairs.     

Tolerance to habitat fragmentation influences the colonization of new habitat by forest birds

We examined the relationship between the ability of bird species to persist in fragmented forests and their ability to colonize new forest habitat. Using a long-term data set on the colonization of a forest plantation, we tested the hypothesis that bird species tolerant to habitat fragmentation would detect and colonize the new habitat faster than intolerant species. The forest plantation under study is situated on an area of land reclaimed from the sea (a polder) in the central part of The Netherlands. We constructed an index of tolerance to habitat fragmentation and included it as a predictor variable in a set of three logistic regression models that compared the probability of colonization over four consecutive time periods. After controlling statistically for the effects of regional incidence, preferred habitat and life-history characteristics, there was a significant effect of tolerance to fragmentation on the ability of species to colonize the plantation, and a marginal effect on the timing of colonization. We then examined the effect of the same index of tolerance to fragmentation on colonization patterns over a larger spatial scale. Multivariate regression models showed that the proportion of three polders of different ages occupied by forest bird species was dependent upon the regional incidence of a species, its preferred habitat and its tolerance to fragmentation. The results support the hypothesis that species tolerant to habitat fragmentation detect and colonize new habitat faster than those intolerant to habitat fragmentation.     

Odor-mediated aggregation enhances the colonization ability of Drosophila melanogaster

Animal aggregation is a general phenomenon in ecological systems. Aggregations are generally considered as an evolutionary advantageous state in which members derive the benefits of mate choice and protection against natural enemies, balanced by the costs of limiting resources and intraspecific competition. Many insects use chemical information to find conspecifics and to form aggregations. In this study, we describe a spatio-temporal simulation model designed to explore and quantify the effects of the strength of chemical attraction, on the colonization ability of a fruit fly (Drosophila melanogaster) population. We found that the use of infochemicals is crucial for colonizing an area. Fruit flies subject to an Allee effect that are unable to respond to chemical information could not successfully colonize the area and went extinct within four generations. This was mainly caused by very high mortality due to the Allee effect. Even when the Allee effect did not play a role, the random dispersing population had more difficulties in colonizing the area and is doomed to extinction in the long run. When fruit flies had the ability to respond to chemical information, they successfully colonized the orchard. This happened faster, for stronger attraction to chemical information. In addition, more fruit flies were able to find the resources and the settlement on the resources was much higher. This resulted in a reduced mortality due to the Allee effect for fruit flies able to respond to chemical information. Odor-mediated aggregation thus enhances the colonization ability of D. melanogaster. Even a weak attraction to chemical information paved the way to successfully colonize the orchard.     

TLR-mediated immune responses in the urinary tract

The urinary tract is one of the most intractable mucosal surfaces for pathogens to colonize. In addition to the natural barriers at this site, potential pathogens have to contend with the vigorous local innate immune system. Several Toll-like receptors (TLRs) have been identified on epithelial cells of the bladder and the kidneys which mediate a variety of powerful immune responses. A common finding among successful uropathogens is their intrinsic ability to suppress TLR-mediated responses. As antibiotic therapy becomes increasingly ineffective, employing boosters of the innate immune system in the urinary tract may become a viable option.     

The trade-off between dispersability and longevity - an important aspect of plant species diversity

Abstract. Local presence of plant species is determined by population colonizations and extinctions. All traits that influence the capacity of individuals to colonize patches and survive within patches, are therefore important for community diversity. Spatial models can explain the coexistence of species provided that the inferior competitor has a greater spatial mobility and thereby can avoid competition. We searched the literature for empirical evidence for such trade-offs and included all available information on correlations between traits associated with the capacity to colonize and traits promoting the ability to survive. A lower reproductive effort of a species is associated with a longer life span and a higher competitive ability. Morphological adaptations for dispersal are less common in species which better tolerate stress, that are better competitors or possess seed dormancy. Such patterns suggest that species that are good survivors may have a limited ability to colonize new patches and vice versa. A negative correlation between dispersability and longevity has important effects on the regional dynamics of single species as well as on the coexistence of species. From a conservation perspective differences in the colonization capacity among species imply that restoration of plant biodiversity must not only focus on conditions within patches, but also consider the spatial arrangement of patches in order to enable plants to bridge gaps in time and space.     

The role of disulfide bond isomerase A (DsbA) of Escherichia coli O157:H7 in biofilm formation and virulence

The role of periplasmic disulfide oxidoreductase DsbA in Shiga toxin-producing Escherichia coli O157:H7 (STEC) was investigated. Deletion of dsbA (DeltadsbA) significantly decreased cell motility and alkaline phosphatase activity in STEC. STEC DeltadsbA also showed greater sensitivity to menadione and under low pH conditions. Significant reductions in surface attachment to both biotic (HT-29 epithelial cells) and abiotic (polystyrene and polyvinyl chloride) surfaces were observed in STEC DeltadsbA. In addition, no biofilm formation was detected in STEC DeltadsbA compared to wild-type cells in glass capillary tubes under continuous flow-culture system conditions. In the nematode model Caenorhabditis elegans-killing assay, the deletion of dsbA in STEC resulted in attenuated virulence compared to wild-type cells. STEC DeltadsbA was also found to have a reduced ability to colonize the nematode gut. These results suggest that DsbA plays important roles in biofilm formation and virulence in STEC cells.     

Role of two G-protein alpha subunits, TgaA and TgaB, in the antagonism of plant pathogens by Trichoderma virens

G-protein alpha subunits are involved in transmission of signals for development, pathogenicity, and secondary metabolism in plant pathogenic and saprophytic fungi. We cloned two G-protein alpha subunit genes, tgaA and tgaB, from the biocontrol fungus Trichoderma virens. tgaA belongs to the fungal Galphai class, while tgaB belongs to the class defined by gna-2 of Neurospora crassa. We compared loss-of-function mutants of tgaA and tgaB with the wild type for radial growth, conidiation, germination of conidia, the ability to overgrow colonies of Rhizoctonia solani and Sclerotium rolfsii in confrontation assays, and the ability to colonize the sclerotia of these pathogens in soil. Both mutants grew as well as the wild type, sporulated normally, did not sporulate in the dark, and responded to blue light by forming a conidial ring. The tgaA mutants germinated by straight unbranched germ tubes, while tgaB mutants, like the wild type, germinated by wavy and highly branched germ tubes. In confrontation assays, both tgaA and tgaB mutants and the wild type overgrew, coiled, and lysed the mycelia of R. solani, but tgaA mutants had reduced ability to colonize S. rolfsii colonies. In the soil plate assay, both mutants parasitized the sclerotia of R. solani, but tgaA mutants were unable to parasitize the sclerotia of S. rolfsii. Thus, tgaA is involved in antagonism against S. rolfsii, but neither G protein subunit is involved in antagonism against R. solani. T. virens, which has a wide host range, thus employs a G-protein pathway in a host-specific manner.     

Germination of Epiphytic Bromeliads in Forests and Coffee Plantations: Microclimate and Substrate Effects

This study explores whether differences in germination ability shape habitat distributions among closely related epiphytic species, particularly in the context of the ability to colonize disturbed habitats. We compared the germination of three epiphytic bromeliad species differing in their ability to colonize secondary arboreal vegetation in natural forests, and in old and young coffee plantations. We asked if germination is related to their distribution within the canopy and between habitats, and the extent to which the substrate (branch) or microclimate affects germination success. Tillandsia viridiflora, a species that is largely restricted to closed forests, had highest germination success in natural forests, Tillandsia juncea, a pioneer species, germinated best in young coffee plantations, and Tillandsia heterophylla, an intermediate species, equally in forests and young plantations. Surprisingly germination rates of all three species were lowest in old plantations. Bryophyte cover on branches had a positive effect on germination of T. viridiflora and T. heterophylla, but T. juncea germination rates were largely independent of climate and substrate. These results show that germination can limit the ability of species to colonize disturbed habitats and also contributes to within‐canopy distribution. Abstract in Spanish is available at http://www.blackwell‐synergy.com/loi/btp .     

Rhizosphere Colonization and Control of Meloidogyne spp. by Nematode-trapping Fungi

The ability of nematode-trapping fungi to colonize the rhizosphere of crop plants has been suggested to be an important factor in biological control of root-infecting nematodes. In this study, rhizosphere colonization was evaluated for 38 isolates of nematode-trapping fungi representing 11 species. In an initial screen, Arthrobotrys dactyloides, A. superba, and Monacrosporium ellipsosporum were most frequently detected in the tomato rhizosphere. In subsequent pot experiments these fungi and the non-root colonizing M. geophyropagum were introduced to soil in a sodium alginate matrix, and further tested both for establishment in the tomato rhizosphere and suppression of root-knot nematodes. The knob-forming M. ellipsosporum showed a high capacity to colonize the rhizosphere both in the initial screen and the pot experiments, with more than twice as many fungal propagules in the rhizosphere as in the root-free soil. However, neither this fungus nor the other nematode-trapping fungi tested reduced nematode damage to tomato plants.