Changes in antimicrobial resistance in fecal bacteria associated with pig transit and holding times at slaughter plants.

Fecal coliforms, fecal streptococci, and antimicrobial resistance (AMR) associated with various pig transit and holding times were investigated at slaughter plants. Changes in the relative abundance of two biotypes of Streptococcus faecium were associated with transit and holding of pigs, although approximately 20% of the isolates were unidentified. The greatest variety of coliforms was isolated from porcine feces after short transit (2 h) or holding (3 h) times and was qualitatively similar to those from pigs on farms. Isolates from pigs with longer average transit or holding times were almost all Escherichia coli (four biotypes). Streptococcal resistance to most antimicrobial agents was significantly greater (P less than 0.05) in isolates from live pigs at slaughter plants than in those from pigs at farms and was apparent after a short transit time (2 h). Streptococci from pigs held an average of 15 h were less resistant to most antimicrobial agents than those from pigs held 3 or 43 h. When compared with short transit times, moderate transit times (6 h) were associated with significantly decreased (P less than 0.05) coliform resistance and decreased resistance transfer but a greater diversity of AMR patterns. Holding pigs overnight (14 h) was associated with lowered coliform resistance to several antimicrobial agents, compared with the resistance of isolates from pigs held 3 or 39 h. A substantial increase (18 to 48%) in the ability to transfer streptomycin resistance was demonstrated in coliforms from pigs held 39 h, when compared with those from pigs held 3 h.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in antimicrobial resistance in fecal bacteria associated with pig transit and holding times at slaughter plants

Fecal coliforms, fecal streptococci, and antimicrobial resistance (AMR) associated with various pig transit and holding times were investigated at slaughter plants. Changes in the relative abundance of two biotypes of Streptococcus faecium were associated with transit and holding of pigs, although approximately 20% of the isolates were unidentified. The greatest variety of coliforms was isolated from porcine feces after short transit (2 h) or holding (3 h) times and was qualitatively similar to those from pigs on farms. Isolates from pigs with longer average transit or holding times were almost all Escherichia coli (four biotypes). Streptococcal resistance to most antimicrobial agents was significantly greater (P less than 0.05) in isolates from live pigs at slaughter plants than in those from pigs at farms and was apparent after a short transit time (2 h). Streptococci from pigs held an average of 15 h were less resistant to most antimicrobial agents than those from pigs held 3 or 43 h. When compared with short transit times, moderate transit times (6 h) were associated with significantly decreased (P less than 0.05) coliform resistance and decreased resistance transfer but a greater diversity of AMR patterns. Holding pigs overnight (14 h) was associated with lowered coliform resistance to several antimicrobial agents, compared with the resistance of isolates from pigs held 3 or 39 h. A substantial increase (18 to 48%) in the ability to transfer streptomycin resistance was demonstrated in coliforms from pigs held 39 h, when compared with those from pigs held 3 h.(ABSTRACT TRUNCATED AT 250 WORDS)     

Metabolomics Analysis Identifies Intestinal Microbiota-Derived Biomarkers of Colonization Resistance in Clindamycin-Treated Mice

Background

The intestinal microbiota protect the host against enteric pathogens through a defense mechanism termed colonization resistance. Antibiotics excreted into the intestinal tract may disrupt colonization resistance and alter normal metabolic functions of the microbiota. We used a mouse model to test the hypothesis that alterations in levels of bacterial metabolites in fecal specimens could provide useful biomarkers indicating disrupted or intact colonization resistance after antibiotic treatment.

Methods

To assess in vivo colonization resistance, mice were challenged with oral vancomycin-resistant Enterococcus or Clostridium difficile spores at varying time points after treatment with the lincosamide antibiotic clindamycin. For concurrent groups of antibiotic-treated mice, stool samples were analyzed using quantitative real-time polymerase chain reaction to assess changes in the microbiota and using non-targeted metabolic profiling. To assess whether the findings were applicable to another antibiotic class that suppresses intestinal anaerobes, similar experiments were conducted with piperacillin/tazobactam.

Results

Colonization resistance began to recover within 5 days and was intact by 12 days after clindamycin treatment, coinciding with the recovery bacteria from the families Lachnospiraceae and Ruminococcaceae, both part of the phylum Firmicutes. Clindamycin treatment caused marked changes in metabolites present in fecal specimens. Of 484 compounds analyzed, 146 (30%) exhibited a significant increase or decrease in concentration during clindamycin treatment followed by recovery to baseline that coincided with restoration of in vivo colonization resistance. Identified as potential biomarkers of colonization resistance, these compounds included intermediates in carbohydrate or protein metabolism that increased (pentitols, gamma-glutamyl amino acids and inositol metabolites) or decreased (pentoses, dipeptides) with clindamycin treatment. Piperacillin/tazobactam treatment caused similar alterations in the intestinal microbiota and fecal metabolites.

Conclusions

Recovery of colonization resistance after antibiotic treatment coincided with restoration of several fecal bacterial metabolites. These metabolites could provide useful biomarkers indicating intact or disrupted colonization resistance during and after antibiotic treatment.     

Induced terpene accumulation in Norway spruce inhibits bark beetle colonization in a dose-dependent manner

Background

Tree-killing bark beetles (Coleoptera, Scolytinae) are among the most economically and ecologically important forest pests in the northern hemisphere. Induction of terpenoid-based oleoresin has long been considered important in conifer defense against bark beetles, but it has been difficult to demonstrate a direct correlation between terpene levels and resistance to bark beetle colonization.

Methods

To test for inhibitory effects of induced terpenes on colonization by the spruce bark beetle Ips typographus (L.) we inoculated 20 mature Norway spruce Picea abies (L.) Karsten trees with a virulent fungus associated with the beetle, Ceratocystis polonica (Siem.) C. Moreau, and investigated induced terpene levels and beetle colonization in the bark.

Results

Fungal inoculation induced very strong and highly variable terpene accumulation 35 days after inoculation. Trees with high induced terpene levels (n = 7) had only 4.9% as many beetle attacks (5.1 vs. 103.5 attacks m⁻²) and 2.6% as much gallery length (0.029 m m⁻² vs. 1.11 m m⁻²) as trees with low terpene levels (n = 6). There was a highly significant rank correlation between terpene levels at day 35 and beetle colonization in individual trees. The relationship between induced terpene levels and beetle colonization was not linear but thresholded: above a low threshold concentration of ∼100 mg terpene g⁻¹ dry phloem trees suffered only moderate beetle colonization, and above a high threshold of ∼200 mg terpene g⁻¹ dry phloem trees were virtually unattacked.

Conclusion/Significance

This is the first study demonstrating a dose-dependent relationship between induced terpenes and tree resistance to bark beetle colonization under field conditions, indicating that terpene induction may be instrumental in tree resistance. This knowledge could be useful for developing management strategies that decrease the impact of tree-killing bark beetles.     

The characteristics of the colonization resistance of the intestines in conventional animals with a history of clinical death]

After modeling the terminal state caused by the acute loss of blood in rats their intestinal microflora was studied, as was their resistance to colonization. Decreased resistance to colonization was registered early after resuscitation (up to 3 days), which was confirmed by the translocation of bacteria into internal organs, decreased number of lactobacilli in the contents of the small intestine and elevated level of enterobacteria in the intestine. Disturbances in resistance to colonization was also manifested by prolonged colonization of the digestive tract of the resuscitated animals by Escherichia coli indicator strain K12pSS-120 carrying Shigella sonnei (phase I) invasiveness plasmid.     

Gnotobiotic studies to determine the colonization resistance of the intestines

The literature data and the results of the author studies on determination of intestine colonization resistance are presented. The mechanisms of the colonization resistance defined by the macroorganism factors and representatives of indigenic microflora are discussed. The results of the experiments with animal gnotobiotes aimed at elucidating new aspects of the colonization resistance mechanism: antagonistic interrelations between pathogenic and nonpathogenic bacteria and the role of transitory microflora, factors lowering the colonization resistance are presented. The up-to-date methods for testing the colonization resistance and the ways for its increasing are indicated.     

A Monogenic Dominant Resistance of Tomato to Bacterial Wilt in Hawaii 7996 is Associated with Plant Colonization by Pseudomonas solanacearum

Hawaii 7996, a tomato cultivar resistant to bacterial wilt caused by P. solanacearum was crossed with Floradel, a susceptible cultivar and the F1 and F2 seeds were obtained. Inoculated plants were tested in the field for bacterial wilt resistance and colonization by P. solanacearum. The F1 did not wilt and a significant 3:1 segregation for non wilting: wilting was observed in the F2, indicating a monogenic dominant resistance in Hawaii 7996. In the F2 and in Hawaii 7996, resistance was, associated to the limitation of bacterial spread in the stem. The degree of resistance of Floradel, the F2 and Hawaï 7996 was correlated to colonization at midstem. The usefulness of plant colonization criteria for breeding programs is discussed.     

Altering host resistance to infections through microbial transplantation

Host resistance to bacterial infections is thought to be dictated by host genetic factors. Infections by the natural murine enteric pathogen Citrobacter rodentium (used as a model of human enteropathogenic and enterohaemorrhagic E. coli infections) vary between mice strains, from mild self-resolving colonization in NIH Swiss mice to lethality in C3H/HeJ mice. However, no clear genetic component had been shown to be responsible for the differences observed with C. rodentium infections. Because the intestinal microbiota is important in regulating resistance to infection, and microbial composition is dependent on host genotype, it was tested whether variations in microbial composition between mouse strains contributed to differences in "host" susceptibility by transferring the microbiota of resistant mice to lethally susceptible mice prior to infection. Successful transfer of the microbiota from resistant to susceptible mice resulted in delayed pathogen colonization and mortality. Delayed mortality was associated with increased IL-22 mediated innate defense including antimicrobial peptides Reg3γ and Reg3β, and immunono-neutralization of IL-22 abrogated the beneficial effect of microbiota transfer. Conversely, depletion of the native microbiota in resistant mice by antibiotics and transfer of the susceptible mouse microbiota resulted in reduced innate defenses and greater pathology upon infection. This work demonstrates the importance of the microbiota and how it regulates mucosal immunity, providing an important factor in susceptibility to enteric infection. Transfer of resistance through microbial transplantation (bacteriotherapy) provides additional mechanisms to alter "host" resistance, and a novel means to alter enteric infection and to study host-pathogen interactions.     

The intestinal microbiota plays a role in Salmonella-induced colitis independent of pathogen colonization

The intestinal microbiota is composed of hundreds of species of bacteria, fungi and protozoa and is critical for numerous biological processes, such as nutrient acquisition, vitamin production, and colonization resistance against bacterial pathogens. We studied the role of the intestinal microbiota on host resistance to Salmonella enterica serovar Typhimurium-induced colitis. Using multiple antibiotic treatments in 129S1/SvImJ mice, we showed that disruption of the intestinal microbiota alters host susceptibility to infection. Although all antibiotic treatments caused similar increases in pathogen colonization, the development of enterocolitis was seen only when streptomycin or vancomycin was used; no significant pathology was observed with the use of metronidazole. Interestingly, metronidazole-treated and infected C57BL/6 mice developed severe pathology. We hypothesized that the intestinal microbiota confers resistance to infectious colitis without affecting the ability of S. Typhimurium to colonize the intestine. Indeed, different antibiotic treatments caused distinct shifts in the intestinal microbiota prior to infection. Through fluorescence in situ hybridization, terminal restriction fragment length polymorphism, and real-time PCR, we showed that there is a strong correlation between the intestinal microbiota composition before infection and susceptibility to Salmonella-induced colitis. Members of the Bacteroidetes phylum were present at significantly higher levels in mice resistant to colitis. Further analysis revealed that Porphyromonadaceae levels were also increased in these mice. Conversely, there was a positive correlation between the abundance of Lactobacillus sp. and predisposition to colitis. Our data suggests that different members of the microbiota might be associated with S. Typhimurium colonization and colitis. Dissecting the mechanisms involved in resistance to infection and inflammation will be critical for the development of therapeutic and preventative measures against enteric pathogens.     

Invasiveness of Avirulent Strains of Pseudomonas solanacearum in Tomato Cultivars, Resistant or Susceptible to Bacterial Wilt

Tomato plant colonization by avirulent strains of Pseudomonas solanacearum was studied. The strain 8217R a spontaneous rough mutant was able to penetrate into roots by nature opennings and colonize plants at high levels. The strain 8173, a hrp^- engineered fluidal mutant, colonized the plants to a lesser extent than 8217R. There was no difference in colonization observed at taproots and hypocotyls between resistant and susceptible cultivars with the avirulent strains 8217R and 8173, contrary to the plant-virulent strain interaction. Results indicated that the bacterial spread was limited in the two cultivars regardless of their resistance or susceptibility and suggested the existence of a host defence mechanism, limiting the bacterial spread.     

Distinct but Spatially Overlapping Intestinal Niches for Vancomycin-Resistant Enterococcus faecium and Carbapenem-Resistant Klebsiella pneumoniae

Antibiotic resistance among enterococci and γ-proteobacteria is an increasing problem in healthcare settings. Dense colonization of the gut by antibiotic-resistant bacteria facilitates their spread between patients and also leads to bloodstream and other systemic infections. Antibiotic-mediated destruction of the intestinal microbiota and consequent loss of colonization resistance are critical factors leading to persistence and spread of antibiotic-resistant bacteria. The mechanisms underlying microbiota-mediated colonization resistance remain incompletely defined and are likely distinct for different antibiotic-resistant bacterial species. It is unclear whether enterococci or γ-proteobacteria, upon expanding to high density in the gut, confer colonization resistance against competing bacterial species. Herein, we demonstrate that dense intestinal colonization with vancomycin-resistant Enterococcus faecium (VRE) does not reduce in vivo growth of carbapenem-resistant Klebsiella pneumoniae. Reciprocally, K. pneumoniae does not impair intestinal colonization by VRE. In contrast, transplantation of a diverse fecal microbiota eliminates both VRE and K. pneumoniae from the gut. Fluorescence in situ hybridization demonstrates that VRE and K. pneumoniae localize to the same regions in the colon but differ with respect to stimulation and invasion of the colonic mucus layer. While VRE and K. pneumoniae occupy the same three-dimensional space within the gut lumen, their independent growth and persistence in the gut suggests that they reside in distinct niches that satisfy their specific in vivo metabolic needs.     

Identification of pneumococcal colonization determinants in the stringent response pathway facilitated by genomic diversity

Background

Understanding genetic determinants of a microbial phenotype generally involves creating and comparing isogenic strains differing at the locus of interest, but the naturally existing genomic and phenotypic diversity of microbial populations has rarely been exploited. Here we report use of a diverse collection of 616 carriage isolates of Streptococcus pneumoniae and their genome sequences to help identify a novel determinant of pneumococcal colonization.

Results

A spontaneously arising laboratory variant (SpnYL101) of a capsule-switched TIGR4 strain (TIGR4:19F) showed reduced ability to establish mouse nasal colonization and lower resistance to non-opsonic neutrophil-mediated killing in vitro, a phenotype correlated with in vivo success. Whole genome sequencing revealed 5 single nucleotide polymorphisms (SNPs) affecting 4 genes in SpnYL101 relative to its ancestor. To evaluate the effect of variation in each gene, we performed an in silico screen of 616 previously published genome sequences to identify pairs of closely-related, serotype-matched isolates that differ at the gene of interest, and compared their resistance to neutrophil-killing. This method allowed rapid examination of multiple candidate genes and found phenotypic differences apparently associated with variation in SP_1645, a RelA/ SpoT homolog (RSH) involved in the stringent response. To establish causality, the alleles corresponding to SP_1645 were switched between the TIGR4:19F and SpnYL101. The wild-type SP_1645 conferred higher resistance to neutrophil-killing and competitiveness in mouse colonization. Using a similar strategy, variation in another RSH gene (TIGR4 locus tag SP_1097) was found to alter resistance to neutrophil-killing.

Conclusions

These results indicate that analysis of naturally existing genomic diversity complements traditional genetics approaches to accelerate genotype-phenotype analysis.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1573-6) contains supplementary material, which is available to authorized users.     

Effects of colonization of a bacterial endophyte,Azospirillum sp. B510, on disease resistance in tomato

A plant growth-promoting bacteria, Azospirillum sp. B510, isolated from rice, can enhance growth and yield and induce disease resistance against various types of diseases in rice. Because little is known about the interaction between other plant species and this strain, we have investigated the effect of its colonization on disease resistance in tomato plants. Treatment with this strain by soil-drenching method established endophytic colonization in root tissues in tomato plant. The endophytic colonization with this strain-induced disease resistance in tomato plant against bacterial leaf spot caused by Pseudomonas syringae pv. tomato and gray mold caused by Botrytis cinerea. In Azospirillum-treated plants, neither the accumulation of SA nor the expression of defense-related genes was observed. These indicate that endophytic colonization with Azospirillum sp. B510 is able to activate the innate immune system also in tomato, which does not seem to be systemic acquired resistance.     

The social structure of microbial community involved in colonization resistance

It is well established that host-associated microbial communities can interfere with the colonization and establishment of microbes of foreign origins, a phenomenon often referred to as bacterial interference or colonization resistance. However, due to the complexity of the indigenous microbiota, it has been extremely difficult to elucidate the community colonization resistance mechanisms and identify the bacterial species involved. In a recent study, we have established an in vitro mice oral microbial community (O-mix) and demonstrated its colonization resistance against an Escherichia coli strain of mice gut origin. In this study, we further analyzed the community structure of the O-mix by using a dilution/regrowth approach and identified the bacterial species involved in colonization resistance against E. coli. Our results revealed that, within the O-mix there were three different types of bacterial species forming unique social structure. They act as ‘Sensor'', ‘Mediator'' and ‘Killer'', respectively, and have coordinated roles in initiating the antagonistic action and preventing the integration of E. coli. The functional role of each identified bacterial species was further confirmed by E. coli-specific responsiveness of the synthetic communities composed of different combination of the identified players. The study reveals for the first time the sophisticated structural and functional organization of a colonization resistance pathway within a microbial community. Furthermore, our results emphasize the importance of ‘Facilitation'' or positive interactions in the development of community-level functions, such as colonization resistance.     

Community diversity outweighs effect of warming on plant colonization

Abiotic environmental change, local species extinctions and colonization of new species often co‐occur. Whether species colonization is driven by changes in abiotic conditions or reduced biotic resistance will affect community functional composition and ecosystem management. We use a grassland experiment to disentangle effects of climate warming and community diversity on plant species colonization. Community diversity had dramatic impacts on the biomass, richness and traits of plant colonists. Three times as many species colonized the monocultures than the high diversity 17 species communities (~30 vs. 10 species), and colonists collectively produced 10 times as much biomass in the monocultures than the high diversity communities (~30 vs. 3 g/m²). Colonists with resource‐acquisitive strategies (high specific leaf area, light seeds, short heights) accrued more biomass in low diversity communities, whereas species with conservative strategies accrued most biomass in high diversity communities. Communities with higher biomass of resident C4 grasses were more resistant to colonization by legume, nonlegume forb and C3 grass colonists, but not by C4 grass colonists. Compared with effects of diversity, 6 years of 3°C‐above‐ambient temperatures had little impact on plant colonization. Warmed subplots had ~3 fewer colonist species than ambient subplots and selected for heavier seeded colonists. They also showed diversity‐dependent changes in biomass of C3 grass colonists, which decreased under low diversity and increased under high diversity. Our findings suggest that species colonization is more strongly affected by biotic resistance from residents than 3°C of climate warming. If these results were extended to invasive species management, preserving community diversity should help limit plant invasion, even under climate warming.     

Plant diversity and community history shift colonization success from early- to mid-successional species

Aims Functional traits are supposed to play an important role in determining the colonization success of new species into established communities. Short-term experimental studies have documented higher resistance of more diverse grasslands against colonization by new species. However, little is known about which traits colonizers should have to successfully invade diverse plant communities in the longer term and how community history may modify the resistance of diverse communities against colonization.Methods In a grassland biodiversity experiment (Jena Experiment) established with different species richness (SR; 1, 2, 4, 8 and 16) and functional group (FG) number and composition (1 to 4; legumes, grasses, small herbs, tall herbs), we studied colonization of naturally dispersed species in split-plots (i) with different duration of weeding (never weeded, weeded for 3 or 6 years and then un-weeded for 1 year) and (ii) with different duration of colonization (7 years, 4 years and 1 year after cessation of weeding).Important findings Resistance against colonization by new species declined with increased duration of weeding (on average 13, 17 and 22 colonizer species in 1-, 4- and 7-year-old communities, respectively). Communities established at low diversity accumulated more colonizer species with a longer duration of weeding than more diverse communities. Duration of colonization had only small effects on the number of colonizer species. Colonizers with early successional traits, i.e. annual life cycle, reproduction by seeds, small seeds, long-lived seeds and an earlier start of a longer flowering period, were favoured in species-poor newly established experimental plant communities (short duration of weeding) and early after cessation of weeding (short duration of colonization). A change from early- to mid-successional traits, i.e. taller growth, perennial life cycle, vegetative reproduction, characterized colonization at increased plant diversity and in communities with legumes or without grasses. Legume absence/grass presence and increased duration of weeding led to a shift in colonizer strategies from rapid nutrient uptake and cycling (higher specific leaf area) to nutrient retention and symbiotic N 2 fixation. Our study shows that non-random trait spectra of naturally dispersed colonizers encompass trade-offs between different functions (reproduction, persistence, growth) reflected in a change from early- to mid-successional traits at increasing plant diversity, with a longer duration of weeding and a longer time of colonization.     

The effect of thyrotropin releasing hormone and morphine on gastrointestinal transit

The effect of thyrotropin releasing hormone (TRH) alone and in combination with morphine on the gastrointestinal transit was investigated by using the charcoal meal test in mice. The intraperitoneal (IP) administration of TRH decreased the transit when given in a dose of 1.0 mg/kg 10 min prior to the meal. The intracerebroventricular (ICV) administration of TRH (10 μg/mouse) also inhibited the transit when given just prior to the charcoal meal. Subcutaneous (SC) administration of morphine (5, 10 and 20 mg/kg) inhibited gastrointestinal transit in a dose dependent manner. When TRH (1, 3 and 10 mg/kg, IP as well as 0.3 μg, ICV) which had no effect on the transit by itself was combined with morphine (10 mg/kg, SC), an enhancement in the inhibition of the transit was observed. TRH-induced inhibition of the transit was antagonized by naloxone (0.1 mg/kg, SC). It is concluded that TRH inhibits gastrointestinal transit in the mouse possibly via the opiate receptor system.     

The reactions of susceptible and resistant tomato cultivars to strains of Verticillium albo-atrum

Three strains of Verticillium albo-atrum causing severe wilt of tomato (T), progressive (Hp ) and fluctuating (HF) wilt of hop, were inoculated through the roots of four tomato cultivars at different inoculum concentrations. Symptoms were assessed visually 42 days after inoculation, and quantitatively on the change in total leaf area compared with controls. Distribution of mycelium and tyloses was determined by sections at 2 cm intervals of root, stem and petiole. Cultivars Loran Blood and Moscow showed resistance to disease expression at all levels of inoculum concentration with the T strain. Bonny Best and Potentate were both susceptible to this strain, but whereas in Potentate, disease severity increased from mild to severe with increase in inoculum concentration, Bonny Best was severely diseased at the lowest level of inoculum. All cultivars showed some susceptibility to the HP and HF strains; the ‘resistance’ of Loran Blood and Moscow was no longer apparent and Bonny Best was most severely affected. The relative susceptibilities to the strains were HF Bonney Best > Loran Blood > Potentate > Moscow, HP Bonny Best > Loran Blood, Moscow > Potentate, T Bonny Best > Potentate > Loran Blood, Moscow. In general, vascular colonization was less in the cultivars Loran Blood and Moscow with all three fungal strains at io⁵propagules/ml level of inoculum, but this was not always correlated with an increase in disease severity. With the exception of the host-pathogen combinations Bonny Best/T, Bonny Best/HF, Potentate/T and Moscow/T, increasing the inoculum concentration to 10⁷propagules/ml increased disease severity but had little or no effect of increasing vascular colonization. In Bonny Best/T, Bonny Best/HF and Potentate/T vascular colonization was reduced with the higher level of inoculum. Moscow showed complete resistance to symptom expression and little vascular colonization with the T strain at 10⁵prop./ml. At 10⁷prop./ml resistance was maintained but there was very extensive growth of mycelium in the vessels. Tylosis resulted from an interaction of host, fungal strain and the level of inoculum and was not always correlated with the degree of vascular colonization. Contrary to previous reports the resistant varieties Loran Blood and Moscow developed acute disease symptoms after inoculation with HP and HF and these were associated with a high level of tylosis rather than mycelial growth. Tylosis and disease severity but not mycelial growth increased with higher levels of inoculum. The results suggested that susceptibility to Verticillium wilt was a complex response depending on host cultivar, fungal strain and the initial inoculum concentration. In some cultivar-pathogen combinations susceptibility was directly proportional to the amount of mycelium present in the vessels, while in others a physiological resistance mechanism independent of the degree of colonization appeared to operate. In a third category, increased disease development rather than resistance was associated with high levels of tylosis.     

Nondigestible Oligosaccharides Enhance Bacterial Colonization Resistance against Clostridium difficile In Vitro

Clostridium difficile is the principal etiologic agent of pseudomembranous colitis and is a major cause of nosocomial antibiotic-associated diarrhea. A limited degree of success in controlling C. difficile infection has been achieved by using probiotics; however, prebiotics can also be used to change bacterial community structure and metabolism in the large gut, although the effects of these carbohydrates on suppression of clostridial pathogens have not been well characterized. The aims of this study were to investigate the bifidogenicity of three nondigestible oligosaccharide (NDO) preparations in normal and antibiotic-treated fecal microbiotas in vitro and their abilities to increase barrier resistance against colonization by C. difficile by using cultural and molecular techniques. Fecal cultures from three healthy volunteers were challenged with a toxigenic strain of C. difficile, and molecular probes were used to monitor growth of the pathogen, together with growth of bifidobacterial and bacteroides populations, over a time course. Evidence of colonization resistance was assessed by determining viable bacterial counts, short-chain fatty acid formation, and cytotoxic activity. Chemostat studies were then performed to determine whether there was a direct correlation between bifidobacteria and C. difficile suppression. NDO were shown to stimulate bifidobacterial growth, and there were concomitant reductions in C. difficile populations. However, in the presence of clindamycin, activity against bifidobacteria was augmented in the presence of NDO, resulting in a further loss of colonization resistance. In the absence of clindamycin, NDO enhanced colonization resistance against C. difficile, although this could not be attributed to bifidobacterium-induced inhibitory phenomena.     

The Intestinal Microbiota Influences Campylobacter jejuni Colonization and Extraintestinal Dissemination in Mice

Campylobacter jejuni is a leading cause of human foodborne gastroenteritis worldwide. The interactions between this pathogen and the intestinal microbiome within a host are of interest as endogenous intestinal microbiota mediates a form of resistance to the pathogen. This resistance, termed colonization resistance, is the ability of commensal microbiota to prevent colonization by exogenous pathogens or opportunistic commensals. Although mice normally demonstrate colonization resistance to C. jejuni, we found that mice treated with ampicillin are colonized by C. jejuni, with recovery of Campylobacter from the colon, mesenteric lymph nodes, and spleen. Furthermore, there was a significant reduction in recovery of C. jejuni from ampicillin-treated mice inoculated with a C. jejuni virulence mutant (ΔflgL strain) compared to recovery of mice inoculated with the C. jejuni wild-type strain or the C. jejuni complemented isolate (ΔflgL/flgL). Comparative analysis of the microbiota from nontreated and ampicillin-treated CBA/J mice led to the identification of a lactic acid-fermenting isolate of Enterococcus faecalis that prevented C. jejuni growth in vitro and limited C. jejuni colonization of mice. Next-generation sequencing of DNA from fecal pellets that were collected from ampicillin-treated CBA/J mice revealed a significant decrease in diversity of operational taxonomic units (OTUs) compared to that in control (nontreated) mice. Taken together, we have demonstrated that treatment of mice with ampicillin alters the intestinal microbiota and permits C. jejuni colonization. These findings provide valuable insights for researchers using mice to investigate C. jejuni colonization factors, virulence determinants, or the mechanistic basis of probiotics.