<p>The crossing of host barriers (intestinal, blood-brain, and placental) is a critical step for systemic infections caused by entero-invasive pathogens. In the intestine, the epithelial cells are the first line of defense against enteric pathogens. <i>Listeria monocytogenes</i> is a facultative-intracellular foodborne pathogen that first crosses the intestinal barrier to cause a systemic infection. However, the underlying mechanism is not well understood.</p><p><br></p> <p>We demonstrate that <i>Listeria</i> adhesion protein (LAP) promotes the translocation of <i>L. monocytogenes </i>across the intestinal barrier in mouse models (A/J and C57BL/6). Relative to the wild-type (WT; serotype 4b) or the isogenic bacterial invasion protein Internalin A mutant (Δ<i>inlA</i>) strain, the <i>lap^─</i> strain showed significant defect in translocation across the intestinal barrier and colonization of the mesenteric-lymph nodes, liver and spleen in the early phase of infection (24 h and 48 h). LAP induces intestinal epithelial barrier dysfunction for increased translocation as evidenced by increased permeability to 4-kDa FITC-dextran (FD4), a marker of paracellular permeability, in the serum and urine of WT and Δ<i>inlA</i>- infected mice and across Caco-2 cell barrier, but not the <i>lap^─</i> mutant strain. Microscopic examination confirmed localization of the WT and Δ<i>inlA</i> strains in the tight junction, a crucial barrier of intestinal paracellular permeability, in the mouse ileal tissue but the <i>lap^─</i> strain remained confined in the lumen. LAP also upregulates TNF-α and IL-6 in intestinal epithelia of mice and in Caco-2 cells for increased permeability. </p><p><br></p> <p>Investigation of the underlying molecular mechanisms of LAP-mediated increase in intestinal permeability by using <i>lap^─</i> mutant strain, purified LAP and shRNA-mediated Hsp60 suppression, we demonstrate that LAP interacts with its host receptor, Hsp60, and activates the canonical NF-κB signaling, which in turn facilitates myosin light-chain kinase (MLCK)-mediated opening of the epithelial barrier via the cellular redistribution of major epithelial junctional proteins claudin-1, occludin, and E-cadherin. Pharmacological inhibition of NF-κB or MLCK in cells or genetic ablation of MLCK in mice (C57BL/6) prevents mislocalization of epithelial junctional proteins, intestinal permeability and <i>L. monocytogenes</i> translocation across the intestinal barrier.</p> <p><br></p><p>Furthermore, LAP also promotes <i>L. monocytogenes </i>translocation across the intestinal barrier and systemic dissemination in a Mongolian gerbil that are permissive to the bacterial invasion proteins; InlA-and InlB-mediated pathways; similar to that in humans. We show a direct LAP-dependent and InlA-independent pathway<i> </i>for <i>L. monocytogenes</i> paracellular translocation across the intestinal epithelial cells that do not express luminally accessible E-cadherin. Additionally, we show a functional InlA/E-cadherin interaction pathway that aids <i>L. monocytogenes</i> translocation by targeting cells with luminally accessible E-cadherin such as cells at the site of epithelial cell extrusion, epithelial folds and mucus-expelling goblet cells. Thus, <i>L. monocytogenes</i> uses LAP to exploit epithelial innate defense in the early phase of infection to cross the intestinal epithelial barrier, independent of other invasion proteins.</p><p><br></p> <p>This work fills a critical gap in our understanding of <i>L. monocytogenes </i>pathogenesis and sheds light to the complex interplay between host-pathogen interactions for bacterial crossing of the crucial intestinal barrier.</p> <br>

Le parasitisme est de plus en plus considéré comme un facteur écologique pouvant créer des variations dans le comportement des individus. Toutefois, la direction de causalité entre le comportement et le parasitisme reste incertaine. Les infections expérimentales sont le plus souvent réalisées en laboratoire, limitant les inférences écologiques. À l’aide d’une infection expérimentale semi-naturelle, nous avons infecté avec succès des crapets-soleils (Lepomis gibbosus) dans un lac où ils ont été exposés à une variété de vers parasites (trématodes, cestodes), permettant d’examiner les effets de la co-infection naturelle chez les hôtes. Nous avons mesuré la témérité, l’exploration et l’activité avant et après l’infection expérimentale. En utilisant une approche bayésienne, nous avons trouvé que les traits de comportement initiaux exercent une forte influence sur la susceptibilité à l’infection : les poissons les plus téméraires et/ou les moins actifs au départ ont acquis une plus grande densité de points noirs (c.-à-d. points noirs visibles sous la peau, les nageoires et dans les muscles du poisson) et de cestodes lors de l’infection. Par ailleurs, nous avons montré que la condition corporelle est réduite par la densité de cestodes, suggérant la débilitation de l’hôte. La condition corporelle était corrélée positivement avec la distance parcourue, quel que soit le statut d’infection individuel. Nous avons également trouvé une relation négative entre la distance parcourue après l’infection et la densité de trématodes, suggérant que l’infection causant la maladie des points noirs diminue le niveau d’activité des hôtes. La témérité et l'exploration n'étaient pas affectées par la densité parasitaire ou la condition corporelle. Nous suggérons que la diminution de l’activité est causée par une réponse adaptative de l’hôte, visant à rediriger son énergie pour combattre l’infection. Sachant que les points noirs ont un cycle de vie complexe et que le crapet-soleil est un hôte intermédiaire, ce changement dans le comportement de l’hôte pourrait aider le parasite à compléter sa transmission aux oiseaux-hôtes piscivores en augmentant la prédation sur les poissons infectés. Bien que nous ne puissions confirmer la direction de causalité, nos résultats montrent qu’il existe un lien étroit entre le comportement et le parasitisme. Nous suggérons que deux mécanismes peuvent simultanément agir : le comportement initial des individus influence le risque d’infection, et l’infection peut créer de la variation au niveau de la plasticité comportementale des individus. / Parasitism is increasingly seen as an ecological factor that can create variations in individual behaviour. However, the direction of causality between behaviour and parasitism remains uncertain. Experimental infections are most often conducted in laboratories, limiting ecological inferences. Using a semi-natural experimental infection, we successfully infected pumpkinseed sunfish (Lepomis gibbosus) in a lake where they were exposed to various parasitic worms (trematodes, cestodes), allowing us to examine the effects of natural co-infection in hosts. We measured boldness, exploration, and activity before and after the experimental infection. Using a Bayesian approach, we found that initial behavioural traits strongly influence infection susceptibility: initially bolder and/or less active fish acquired a higher density of black spots (i.e., visible black spots under the skin, fins, and in the fish's muscles) and cestodes during the infection. Additionally, we found that body condition was reduced by cestode density, suggesting host debilitation. Body condition was positively correlated with distance swam, regardless of individual infection status. We also found a negative relationship between distance swam after infection and trematode density, suggesting that infection causing black spot disease decreases host activity levels. Boldness and exploration were not affected by parasite density or body condition. We suggest that a decrease in activity is caused by an adaptive host response to redirect its energy to combat the infection. However, since trematode parasites have a complex life cycle and pumpkinseed sunfish are intermediate hosts, decreases in activity levels following infection may make infected fish more susceptible to predation by piscivorous birds, which is needed for trematodes to complete their life cycles. While we cannot confirm the direction of causality, our results show a close link between behaviour and parasitism. We propose that two mechanisms may simultaneously operate: the initial behaviour of individuals influences their risk of infection, and infection can create variation in behavioural plasticity of individuals.

No description available.

<p>Eastern black walnut (<i>Juglans nigra</i> L.) ranks among the most highly valued timber species in the central hardwood forest and across the world. This valuable tree fills a critical role in native ecosystems as a mast bearing pioneer on mesic sites. Along with other <i>Juglans</i> spp. (Juglandaceae), <i>J. nigra</i> is threatened by thousand cankers disease (TCD), an insect-vectored disease first described in 2009. TCD is caused by the bark beetle <i>Pityophthorus juglandis</i> Blackman (Corthylini) and the phytopathogenic fungus <i>Geosmithia morbida</i> Kol. Free. Ut. & Tiss. (Bionectriaceae). Together, the <i>P. juglandis</i>-<i>G. morbida</i> complex has expanded from its historical range in southwest North America throughout the western United States (U.S.) and Europe. This range expansion has led to widespread mortality among naïve hosts <i>J. nigra</i> and <i>J. regia</i> planted outside their native distributions.</p> <p> The severity of TCD was previously observed to be highest in urban and plantation environments and outside of the host native range. Therefore, the objective of this work was to provide information on biotic and abiotic environmental factors that influence the severity and impact of TCD across the native and non-native range of <i>J. nigra</i> and across different climatic and management regimes. This knowledge would enable a better assessment of the risk posed by TCD and a basis for developing management activities that impart resilience to natural systems. Through a series of greenhouse-, laboratory- and field-based experiments, environmental factors that affect the pathogenicity and/or survival of <i>G. morbida</i> in <i>J. nigra</i> were identified, with a focus on the microbiome, climate, and opportunistic pathogens. A number of potentially important interactions among host, vector, pathogen and the rest of the holobiont of TCD were characterized. The <i>holobiont</i> is defined as the whole multitrophic community of organisms—including <i>J. nigra</i>, microinvertebrates, fungi and bacteria—that interact with one another and with the host.</p> <p>Our findings indicate that interactions among host, vector, pathogen, secondary pathogens, novel microbial communities, and novel abiotic environments modulate the severity of TCD in native, non-native, and managed and unmanaged contexts. Prevailing climatic conditions favor reproduction and spread of <i>G. morbida</i> in the western United States due to the effect of wood moisture content on fungal competition. The microbiome of soils, roots, and stems of trees and seedlings grown outside the host native range harbor distinct, lower-diversity communities of bacteria and fungi compared to the native range, including different communities of beneficial or pathogenic functional groups of fungi. The pathogen <i>G. morbida</i> was also associated with a distinct community of microbes in stems compared to <i>G. morbida</i>-negative trees. The soil microbiome from intensively-managed plantations facilitated positive feedback between <i>G. morbida</i> and a disease-promomting endophytic <i>Fusarium solani</i> species complex sp. in roots of <i>J. nigra</i> seedlings. Finally, the nematode species <i>Bursaphelenchus juglandis</i> associated with <i>P. juglandis</i> synergizes with <i>G. morbida</i> to cause foliar symptoms in seedlings in a shadehouse; conversely, experiments and observations indicated that the nematode species <i>Panagrolaimus</i> sp. and cf. <i>Ektaphelenchus</i> sp. could suppress WTB populations and/or TCD outbreaks.</p> <p>In conclusion, the composition, function, and interactions within the <i>P. juglandis</i> and <i>J. nigra</i> holobiont play important roles in the TCD pathosystem. Managers and conservationists should be aware that novel associations outside the host native range, or in monocultures, intensive nursery production, and urban and low-humidity environments may favor progression of the disease through the effects of associated phytobiomes, nematodes, and climatic conditions on disease etiology. Trees in higher diversity, less intensively managed growing environments within their native range may be more resilient to disease. Moreover, expatriated, susceptible host species (<i>i.e.</i>, <i>J. nigra</i>) growing in environments that are favorable to novel pests or pest complexes (<i>i.e.</i>, the western U.S.) may provide connectivity between emergent forest health threats (<i>i.e.</i>, TCD) and native host populations (<i>i.e.</i>, <i>J. nigra</i> in its native range).</p>