Host ecology and biogeography drive parasite community composition in Atlantic killifishes.

Understanding the mechanisms of parasite community assembly can be confounded by phylogenetic distance among host species. Addressing this requires focusing on parasite communities within closely related taxa. Thus, we took a macroecological approach to examining parasite community structure within Killifish species in the genus Fundulus to disentangle the effects of host phylogeny and ecological variables. We constructed a database of parasite communities within Fundulus species from 15 published and unpublished surveys covering the Atlantic coast of the US and Canada. The database was expanded by sampling sites in underrepresented provinces and states, totaling 10 Fundulus species from 57 unique geographic sites. Univariate analysis of observed parasite species richness among Fundulus populations in the dataset found that latitude, climate type, and salinity were the dominant factors determining parasite species richness. Multivariate analysis found that host species and landscape type were the most important factors in determining the similarity of parasite assemblages. Unexpectedly, parasite species richness decreased in low latitudes, and host phylogenetic distance was not found to be a significant factor in the similarity of parasite communities. These results indicate that commonly reported large-scale drivers of parasite community structure, such as latitude and phylogeny, could have diminished significance at the host genus level relative to host ecology, biogeography, and local landscape factors.

Growth productivity as a determinant of the inoculum effect for bactericidal antibiotics.

Understanding the mechanisms by which populations of bacteria resist antibiotics has implications in evolution, microbial ecology, and public health. The inoculum effect (IE), where antibiotic efficacy declines as the density of a bacterial population increases, has been observed for multiple bacterial species and antibiotics. Several mechanisms to account for IE have been proposed, but most lack experimental evidence or cannot explain IE for multiple antibiotics. We show that growth productivity, the combined effect of growth and metabolism, can account for IE for multiple bactericidal antibiotics and bacterial species. Guided by flux balance analysis and whole-genome modeling, we show that the carbon source supplied in the growth medium determines growth productivity. If growth productivity is sufficiently high, IE is eliminated. Our results may lead to approaches to reduce IE in the clinic, help standardize the analysis of antibiotics, and further our understanding of how bacteria evolve resistance.

Life in the margins: host-parasite relationships in ecological edges.

Transitional zones, such as edge habitat, are key landscapes for investigating biodiversity. "Soft edges" are permeable corridors that hosts can cross, while "hard edges" are impermeable borders that hosts cannot pass. Although pathogen transmission in the context of edges is vital to species conservation, drivers of host-parasite relationships in ecological edges remain poorly understood. Thus, we defined a framework for testing hypotheses of host-parasite interactions in hard and soft edges by (1) characterizing hard and soft edges from both the host and parasite perspectives, (2) predicting the types of parasites that would be successful in each type of edge, and (3) applying our framework to species invasion fronts as an example of host-parasite relationships in a soft edge. Generally, we posited that parasites in soft edges are more likely to be negatively affected by habitat fragmentation than their hosts because they occupy higher trophic levels but parasite transmission would benefit from increased host connectivity. Parasites along hard edges, however, are at higher risk of local extinction due to host population perturbations with limited opportunity for parasite recolonization. We then used these characteristics to predict functional traits that would lead to parasite success along soft and hard edges. Finally, we applied our framework to invasive species fronts to highlight predictions regarding host connectivity and parasite traits in soft edges. We anticipate that our work will promote a more complete discussion of habitat connectivity using a common framework and stimulate empirical research into host-parasite relationships within ecological edges and transitional zones.

Stable isotopes and community surveys reveal differential use of artificial and natural reefs by South Florida fishes.

Artificial reefs may enhance the biological production of reef-associated flora and fauna, but their trophic structure relative to that of natural reefs remains understudied. We assessed trophic relationships by 1) comparing reef fish communities and 2) comparing δ13C and δ15N in 43 fish species from both artificial reef sites and adjacent natural reef tracts in Broward County, Florida. We tested the effect of sampling location (artificial, first, and second reef), general feeding strategy (herbivore, omnivore, invertivore, and carnivore), phylogeny, and standard length on δ13C and δ15N. The reef fish communities of the artificial and natural reef tracts were significantly different; the artificial sites also exhibited more variability. For all samples, δ13C and δ15N ranged from -19.5 to -13.1‰ and 6.7-13.3‰, respectively. Significant effects were detected for both general feeding strategy and phylogeny. Significant differences were also seen in δ13C and δ15N profiles between artificial and natural reefs; however, these changes were primarily driven by differences in fish community structure, rather than by changes in the feeding strategy or trophic relationships of individual fish taxa. The trophic guild invertivore was the only group of fish to demonstrate significant isotopic differences between both reef tracts (inner and outer) and reef types (artificial and natural). The artificial reef may act more as a foraging corridor between the natural first and second reef tracts for omnivores and carnivores. If the function of artificial reefs is to provide additional foraging habitat for fishes, then perhaps more time is needed for the trophically important, infaunal invertebrate community to develop similarly to the natural reef environment.

A potential tradeoff between feeding rate and aversive learning determines intoxication in a Caenorhabditis elegans host-pathogen system.

Despite being the first line of defense against infection, little is known about how host-pathogen interactions determine avoidance. Caenorhabditis elegans can become infected by chemoattractant-producing bacteria through ingestion. The worms can learn to associate these chemoattractants with harm through aversive learning. As a result, the worms will avoid the pathogen. Evolutionary constraints have likely shaped the attraction, intoxication and learning dynamics between bacteria and C. elegans, but these have not been explored. Using bacteria engineered to express an acylhomoserine lactone chemoattractant and a nematicidal protein, we explored how manipulating the amount of attractant produced by the bacteria affects learning and intoxication in mixed stage populations of C. elegans. We found that increasing the production rate of the chemoattractant increased the feeding rate in C. elegans, but decreased the time required for C. elegans to learn to avoid the chemoattractant. Learning generally coincided with a decreased feeding rate. We also observed that the percentage of intoxicated worms was maximized at intermediate production rates of the attractant. We propose that interactions between attractant driven feeding rate and aversive learning are likely responsible for this trend. Our results increase our understanding of behavioral avoidance in C. elegans and have implications in understanding host-pathogen dynamics that shape avoidance.

Validity of the Diplostomoidea and Diplostomida (Digenea, Platyhelminthes) upheld in phylogenomic analysis.

Higher systematics within the Digenea, Carus 1863 have been relatively stable since a phylogenetic analysis of partial nuclear ribosomal markers (rDNA) led to the erection of the Diplostomida Olson, Cribb, Tkach, Bray, and Littlewood, 2003. However, recent mitochondrial (mt) genome phylogenies suggest this order might be paraphyletic. These analyses show members of two diplostomidan superfamilies are more closely related to the Plagiorchiida La Rue, 1957 than to other members of the Diplostomida. A recent phylogeny based on partial cytochrome c oxidase I also indicates one of the groups implicated, the Diplostomoidea Poirier, 1886, is non-monophyletic. To determine if these results were robust to additional taxon sampling, we analyzed mt genomes from seven diplostomoids in three families. To choose between phylogenetic alternatives based on mt genomes and the prior rDNA-based topology, we analyzed hundreds of ultra-conserved genomic elements assembled from shotgun sequencing. The Diplostomida was paraphyletic in the mt genome phylogeny but supported in the ultra-conserved genomic element phylogeny. We speculate this mitonuclear discordance is related to ancient, rapid radiation in the Digenea. Both ultra-conserved genomic elements and mt genomes support the monophyly of the Diplostomoidea and show congruent relationships within it. The Cyathocotylidae Mühling, 1898 are early diverging descendants of a paraphyletic clade of Diplostomidae Poirier, 1886, in which are nested members of the Strigeidae Railliet, 1919; the results support prior suggestions that the Crassiphialinae Sudarikov, 1960 will rise to the family level. Morphological traits of diplostomoid metacercariae appear to be more useful for differentiating clades than those of adults. We describe a new species of Cotylurus Szidat, 1928, resurrect a species of Hysteromorpha Lutz, 1931, and find support for a species of Alaria Schrank, 1788 of contested validity. Complete rDNA operons from seven diplostomoid species are provided as a resource for future studies.

Host Abundance, Sea-Grass Cover, and Temperature Predict Infection Rates of Parasitic Isopods (Bopyridae) on Caridean Shrimp.

Caridean shrimp are critical components of sea-grass communities and occasionally harbor parasitic bopyrid isopods, which can negatively impact their hosts. However, the ecological factors that drive infection rates of parasitic bopyrid isopods in host carideans are poorly known. We examined 43,875 carideans belonging to 6 families and 11 genera from 19 shallow-water localities throughout south Florida. Of these, only 114 shrimp (belonging to 5 genera) were found to be infested with bopyrids (an additional 251 had deformed carapaces consistent with recent infestation). We identified 13 bopyrid species ( Bopyrina abbreviata, Bopyrinella thorii, Eophrixus subcaudalis, Loki circumsaltanus, Metaphrixus carolii, Ovobopyrus alphezemiotes, Parabopyrella lata, Parabopyrella richardsonae, Parabopyriscus stellatus, Capitetragonia alphei, Probopyrus pandalicola, Schizobopyrina urocaridis, and an unidentified Diplophryxus sp.). Bopyrid infection rates were very low throughout the study area, with mean prevalence of 0.26% (range 0.04-1.48%). Furthermore, each isopod species was only ever recovered from a single host genus, suggesting a high degree of genus-level specificity. At the community level, multivariate analyses (RELATE and BVSTEP) indicated that bopyrid community composition was correlated with host community structure, latitude, and temperature, as well as the relative coverage of the sea grasses Thalassia sp. and Syringodium sp. and the alga Penicillus sp. Only 4 parasite taxa were sufficiently abundant to warrant further analysis at the individual taxon level: B. abbreviata, B. thorii, Diplophryxus sp., and P. pandalicola; stepwise regression indicated that bopyrid infection rates were primarily driven by the abundance of their specific hosts, and secondarily by environmental variables such as temperature and depth, as well as algal and sea-grass community composition.

Circumvention of Learning Increases Intoxication Efficacy of Nematicidal Engineered Bacteria.

Synthetic biology holds promise to engineer systems to treat diseases. One critical, yet underexplored, facet of designing such systems is the interplay between the system and the pathogen. Understanding this interplay may be critical to increasing efficacy and overcoming resistance against the system. Using the principles of synthetic biology, we engineer a strain of Escherichia coli to attract and intoxicate the nematode Caenorhabditis elegans. Our bacteria are engineered with a toxin module, which intoxicates the nematode upon ingestion, and an attraction module, which serves to attract and increase the feeding rate of the nematodes. When independently implemented, these modules successfully intoxicate and attract the worms, respectively. However, in combination, the efficacy of our bacteria is significantly reduced due to aversive associative learning in C. elegans. Guided by mathematical modeling, we dynamically regulate module induction to increase intoxication by circumventing learning. Our results detail the creation of a novel nematicidal bacterium that may have application against nematodes, unravel unique constraints on circuit dynamics that are governed by C. elegans physiology, and add to the growing list of design and implementation considerations associated with synthetic biology.

Natural and anthropogenic factors shape metazoan parasite community structure in mummichog (Fundulus heteroclitus) from two estuaries in New Brunswick, Canada.

Previously published multidisciplinary studies in the Miramichi and Bouctouche rivers (New Brunswick, Canada) noted significant changes in fish health parameters, including elevated tissue levels of organic contaminants and a wide range of physiological disturbances, in mummichog Fundulus heteroclitus (L.) from a site on the Miramichi River that received bleached kraft pulpmill and municipal effluent. The present study reports differences in the abundance of individual parasite species, as well as parasite infracommunity and component community composition, in mummichog from both rivers. These differences were evaluated in relation to host (size, condition, immune function, tissue organochlorine contaminant levels) and environmental (faecal coliform counts, salinity, temperature) data derived from the previously published studies. Overall, 18 parasite species were identified, the most common of which were Ascocotyle sp. larv., Ornithodiplostomum sp. larv., Posthodiplostomum sp. larv., and Proteocephalus filicollis (Rudolphi, 1802). There were broad differences in parasite community structure and composition between rivers and within rivers, the most prominent pattern being a pronounced difference between sites in the upper and lower estuary of each river that was likely driven by salinity. Mean infracommunity richness was also positively related to faecal coliforms (considered here as a surrogate measure of eutrophication via municipal sewage), and both were highest at the most polluted site. We noted no other significant relationships. Thus our data suggest that the parasite communities in these two estuaries were primarily structured by large upstream/downstream ecological gradients in salinity, and secondarily by eutrophication due to pollution by municipal and industrial effluents. Overall, our results highlight the value of coordinated multidisciplinary studies for understanding the factors that shape parasite abundance and community structure.

Exposure to a mixture of zinc and copper decreases survival and fecundity of Discocotyle sagittata (Leuckart) parasitizing juvenile Atlantic salmon, Salmo salar L.

We assessed the effects of zinc and copper on freshwater monogenean ectoparasites (Discocotyle sagittata Leuckart) infecting juvenile Atlantic salmon (Salmo salar L.). Exposure to 47 microg/L zinc and 3 microg/L copper reduced survival and fecundity of adult D. sagittata, while egg hatching success was only reduced at high exposure concentrations (2704 microg/L zinc and 164 microg/L copper). Parasitized salmon had decreased plasma chloride, but this was negated in infected fish exposed to metals. No other effects on Atlantic salmon survival and physiology (plasma osmolality, hematocrit) were noted, suggesting that D. sagittata may be more susceptible to metal toxicity than its host fish.

Pollution and parasitism in aquatic animals: a meta-analysis of effect size.

Numerous studies have indicated that aquatic pollution affects parasite populations and communities. However, the responses of individual taxa and functional groups to specific contaminants, and their effect sizes, have yet to be assessed quantitatively. We began by summarizing general trends in the literature, and found that reports of significant changes in parasitism were most commonly observed in response to eutrophication and metal contamination. Among parasite taxa, significant changes were most commonly reported for acanthocephalans, digeneans and microparasites. We then conducted a quantitative meta-analysis of the effects of pollution on parasitism in aquatic animals. We calculated signed standardized effect sizes (as Cohen's d) for all published studies that provided the necessary descriptive statistics, and compared them among major contaminant types (pesticides, hydrocarbons, polychlorinated biphenyls, pulp mill effluent, metals, sewage, eutrophication, and acidification) and parasite taxa (Acanthocephala, Cestoda, Digenea, Monogenea, Nematoda and microparasites). Effect sizes were not significantly different from zero for many parasite/contaminant interactions, and tended to be highly variable within individual taxa. However, consistently strong, significant negative effects were noted in Digenea and Monogenea, especially in response to metal pollution. Significant effect sizes were typically negative, indicating that pollutants have negative effects on parasite populations. Contaminants also had a slightly negative effect on community richness. When parasites were grouped into heteroxenous (with >1 obligatory host in life cycle) vs. monoxenous (1 obligatory host in life cycle) taxa, the latter were more susceptible to a wide range of pollutants. Similarly, directly exposed (external parasites and the free-living transmission stages of internal parasites) and freshwater taxa were more susceptible to a wider range of pollutants than indirectly exposed (internal parasites) and marine taxa. This meta-analysis represented a first attempt to consider the size of the effect of pollution on parasites, and highlighted the potential of susceptible parasite taxa, communities, and functional groups for use in the biological assessment of pollution. For instance, our results indicate that freshwater monogeneans and digeneans are good candidates as potential biological indicators of pollution in aquatic ecosystems.