Collective behavior and virulence arsenal of the fish pathogen Piscirickettsia salmonis in the biofilm realm

Piscirickettsiosis is a fish disease caused by the Gram-negative bacterium Piscirickettsia salmonis. This disease has a high socio-economic impact on the Chilean salmonid aquaculture industry. The bacterium has a cryptic character in the environment and their main reservoirs are yet unknown. Bacterial biofilms represent a ubiquitous mechanism of cell persistence in diverse natural environments and a risk factor for the pathogenesis of several infectious diseases, but their microbiological significance for waterborne veterinary diseases, including piscirickettsiosis, have seldom been evaluated. This study analyzed the in vitro biofilm behavior of P. salmonis LF-89T (genogroup LF-89) and CA5 (genogroup EM-90) using a multi-method approach and elucidated the potential arsenal of virulence of the P. salmonis LF-89T type strain in its biofilm state. P. salmonis exhibited a quick kinetics of biofilm formation that followed a multi-step and highly strain-dependent process. There were no major differences in enzymatic profiles or significant differences in cytotoxicity (as tested on the Chinook salmon embryo cell line) between biofilm-derived bacteria and planktonic equivalents. The potential arsenal of virulence of P. salmonis LF-89T in biofilms, as determined by whole-transcriptome sequencing and differential gene expression analysis, consisted of genes involved in cell adhesion, polysaccharide biosynthesis, transcriptional regulation, and gene mobility, among others. Importantly, the global gene expression profiles of P. salmonis LF-89T were not enriched with virulence-related genes upregulated in biofilm development stages at 24 and 48 h. An enrichment in virulence-related genes exclusively expressed in biofilms was also undetected. These results indicate that early and mature biofilm development stages of P. salmonis LF-89T were transcriptionally no more virulent than their planktonic counterparts, which was supported by cytotoxic trials, which, in turn, revealed that both modes of growth induced important and very similar levels of cytotoxicity on the salmon cell line. Our results suggest that the aforementioned biofilm development stages do not represent hot spots of virulence compared with planktonic counterparts. This study provides the first transcriptomic catalogue to select specific genes that could be useful to prevent or control the (in vitro and/or in vivo) adherence and/or biofilm formation by P. salmonis and gain further insights into piscirickettsiosis pathogenesis

Trophic ecology of the chiton Acanthopleura echinata on Chilean rocky shores Ecología trófica del chitón Acanthopleura echinata en costas rocosas de Chile

Polyplacophorans are common herbivores on rocky shores, but basic aspects of their ecology remain scarcely studied and their role within communities could be more complex than previously considered. Such is the case of Acanthopleura echinata (Barnes), one of the largest and most conspicuous chitons in the world, and at the same time, one of the least known intertidal species in the southeastern Pacific. To improve the basic ecological knowledge of this potentially important intertidal consumer, we studied the diet of A. echinata and its variation among sites of varying levels of coastal upwelling spread over 1000 km along the coast of northern Chile. A seasonal evaluation of diet, body size distribution and density at sites expected to vary in overall nutrient loadings, benthic algal productivity and sea surface temperature, allowed us to examine plasticity in Acantholeura diet and body size. Overall, A. echinata consumed 85 items of algae (64.7 %, mainly fleshy and calcified encrusting thalli) and invertebrates (35.3 %, mainly barnacles). Diet was always dominated by encrusting corallines, although the proportion of algae increased with body size suggesting an ontogenetic variation in feeding habits. Although the number and occurrence frequency of dietary items varied significantly in time and space, there were no consistent seasonal patterns and the dominant items in the diet remained the same at all places. The density of A. echinata showed no significant spatial variation, but its body size and diet breadth correlated positively among sites and both tended to increase with latitude. Our results show that A. echinata is a generalist, polyphagous consumer with a high potential for affecting the space-occupancy dynamics in the intertidal system, and also that among site variation in diet bears no simple relationship with variation in sea surface temperature and upwelling intensity.<br>Los poliplacóforos son herbívoros comunes en costas rocosas, pero los aspectos básicos de su ecología han sido escasamente estudiados y su rol trófico dentro de las comunidades podría ser más complejo de lo considerado hasta ahora. Tal es el caso de Acanthopleura echinata (Barnes), uno de los chitones más grandes y conspicuos en el mundo, pero a la vez una de las especies intermareales menos conocidas del Pacífico sudeste. A fin de mejorar el conocimiento ecológico básico de este consumidor intermareal potencialmente importante, estudiamos la dieta de A. echinata y su variación entre sitios con distintos niveles de surgencia costera distribuidos a través de 1000 km en la costa del norte de Chile. Una evaluación estacional de la dieta, distribución de tamaño corporal y densidad en sitios con diferencias esperables en aporte total de nutrientes, productividad de algas bentónicas y temperatura superficial del mar, permitió examinar la plasticidad de la dieta y el tamaño corporal de Acanthopleura. Globalmente, A. echinata consumió 85 ítemes de algas (64.7 %, principalmente algas incrustantes calcificadas y blandas) e invertebrados (35.3 %, principalmente cirripedios). Su dieta estuvo siempre dominada por coralinas incrustantes, aunque la proporción de algas aumentó con el tamaño corporal sugiriendo una variación ontogenética en sus hábitos tróficos. Aunque el número y frecuencia de ocurrencia de ítemes dietarios variaron significativamente en tiempo y espacio, no hubo patrones estacionales consistentes y los ítemes dominantes en la dieta fueron los mismos en todos los lugares. La densidad de A. echinata no varió significativamente a través del espacio, pero su tamaño corporal y su amplitud dietaria se correlacionaron positivamente entre sitios y ambos tendieron a aumentar con la latitud. Nuestros resultados muestran que A. echinata es un consumidor generalista y polífago con alto potencial para afectar la dinámica de ocupación del espacio en el sistema intermareal, y además que la variación dietaria entre sitios no tiene una relación evidente con la variación en intensidad de surgencia y temperatura superficial del mar

Caracterización trófica del placóforo intermareal Enoplochiton niger en el norte de Chile: variación ambiental y patrones dietarios a nivel local y region Trophic characterization of the intertidal placophoran Enoplochiton niger in northern Chile: environmental variation and dietary patterns at local and regional levels

El rol e impacto de los herbívoros en la trama trófica de comunidades intermareales rocosas podría ser más diverso y complejo de lo considerado hasta ahora, particularmente en el caso de consumidores de mayor tamaño y abundancia como el molusco placóforo Enoplochiton niger. Esta especie es uno de los pastoreadores de mayor tamaño (hasta 20 cm) e importancia ecológica en las costas rocosas del norte de Chile, pero también una de las especies menos conocidas en términos tróficos. Este trabajo presenta una evaluación de los patrones dietarios de E. niger en cuatro comunidades del norte de Chile, distribuidas en 1.000 km de costa y muestreadas estacionalmente entre invierno 2004 y otoño 2006. Además se analizó la relación de su dieta con factores biológicos y físicos, incluyendo el efecto potencial del evento El Niño 2004-2005 ocurrido durante el período de estudio. A nivel regional, el espectro dietario de E. niger abarcó un total de 98 recursos (60 ítemes algales y 38 ítemes invertebrados), y los ítemes más importantes fueron organismos sésiles con formas de crecimiento incrustante o en capa. E. niger mostró una gran amplitud de nicho tanto a nivel regional como local (rango: 20,7-28,0; índice de Levins), con una riqueza dietaria a nivel individual independiente del tamaño corporal. Tanto el número de ítemes dietarios consumidos por individuo como la composición taxonómica de la dieta no mostraron diferencias significativas entre comunidades, pero variaron significativamente entre el período asociado al evento El Niño y el período posterior. No hubo ninguna relación clara entre los patrones dietarios de E. niger y los niveles contrastantes de intensidad de surgencia entre las comunidades estudiadas. Los resultados muestran que E. niger es un consumidor generalista y polífago, y un potencial omnívoro, el cual podría tener un alto impacto sobre los patrones de ocupación de espacio en la comunidad intermareal.<br>The role and impact of herbivores on rocky intertidal food webs could be more complex and diverse than previously considered, particularly in the case of larger and more abundant consumers such as the placophoran mollusc Enoplochiton niger. This species is one of the largest (up to 20 cm) and ecologically most important grazers on rocky shores of northern Chile, but also one of the lesser known species in trophic terms. This work presents an assessment of the dietary patterns of E. niger in four communities of northern Chile, distributed along 1,000 km of coastline and sampled seasonally from winter 2004 to autumn 2006. The analysis included the relationship of its diet with physical and biological factors, and particularly the potential effect of the 2004-2005 El Niño event that occurred during the study period. At a regional level, the dietary spectrum of E. niger comprised a total of 98 food resources (60 algal Ítems and 38 invertebrate Ítems), and the most important Ítems were sessile organisms of encrusting and layer-forming growth habits. E. niger exhibited a very high niche breadth at both local and regional levels (ranging from 20.7 to 28.0; Levins' index), and its dietary richness at the individual level was independent from its body size. Both the number of dietary ítems consumed per individual and the taxonomic composition of the diet did not show statistical differences among communities, but they were significantly different between the El Niño and non-El Niño periods. There was no clear relationship between the dietary patterns of E. niger and the contrasting levels of upwelling intensity among the communities studied. The results show that E. niger is a generalist polyphagous consumer, and a potential omnivore, which could have a high level of impact on space occupancy patterns in the intertidal community

Pre-fibrillar α-synuclein variants with impaired β-structure increase neurotoxicity in Parkinson's disease models

The relation of α-synuclein (αS) aggregation to Parkinson's disease (PD) has long been recognized, but the mechanism of toxicity, the pathogenic species and its molecular properties are yet to be identified. To obtain insight into the function different aggregated αS species have in neurotoxicity in vivo, we generated αS variants by a structure-based rational design. Biophysical analysis revealed that the αS mutants have a reduced fibrillization propensity, but form increased amounts of soluble oligomers. To assess their biological response in vivo, we studied the effects of the biophysically defined pre-fibrillar αS mutants after expression in tissue culture cells, in mammalian neurons and in PD model organisms, such as Caenorhabditis elegans and Drosophila melanogaster. The results show a striking correlation between αS aggregates with impaired β-structure, neuronal toxicity and behavioural defects, and they establish a tight link between the biophysical properties of multimeric αS species and their in vivo function

Protein target highlights in CASP15: Analysis of models by structure providers

We present an in-depth analysis of selected CASP15 targets, focusing on their biological and functional significance. The authors of the structures identify and discuss key protein features and evaluate how effectively these aspects were captured in the submitted predictions. While the overall ability to predict three-dimensional protein structures continues to impress, reproducing uncommon features not previously observed in experimental structures is still a challenge. Furthermore, instances with conformational flexibility and large multimeric complexes highlight the need for novel scoring strategies to better emphasize biologically relevant structural regions. Looking ahead, closer integration of computational and experimental techniques will play a key role in determining the next challenges to be unraveled in the field of structural molecular biology