Microbial communities are indicators of parasite infection status.

Growing evidence suggests that microbiomes have been shaping the evolutionary pathways of macroorganisms for millennia and that these tiny symbionts can influence, and possibly even control, species interactions like host-parasite relationships. Yet, while studies have investigated host-parasites and microbiomes separately, little has been done to understand all three groups synergistically. Here, we collected infected and uninfected Eurypanopeus depressus crab hosts from a coastal North Carolina oyster reef three times over 4 months. Infected crabs demonstrated an external stage of the rhizocephalan parasite, Loxothylacus panopaei. Community analyses revealed that microbial richness and diversity were significantly different among tissue types (uninfected crab, infected crab, parasite externae and parasite larvae) and over time (summer and fall). Specifically, the microbial communities from parasite externae and larvae had similar microbiomes that were consistent through time. Infected crabs demonstrated microbial communities spanning those of their host and parasite, while uninfected crabs showed more distinctive communities with greater variability over time. Microbial communities were also found to be indicators of early-stage infections. Resolving the microbial community composition of a host and its parasite is an important step in understanding the microbiome's role in the host-parasite relationship and determining how this tripartite relationship impacts coevolutionary processes.

Native tube-building polychaete prefers to anchor non-native alga over other macrophytes.

Novel facultative mutualisms that develop between native and non-native ecosystem engineers can lead to the retention of the non-native partner. In some cases, behavior plays an additional, but less understood, role in the development and persistence of mutualisms. In soft-sediment marine habitats along the western Atlantic, the native decorator worm Diopatra cuprea anchors the non-native red alga Gracilaria vermiculophylla to its tube cap in a mutualism. To understand whether the worm's usage of G. vermiculophylla could represent a preference, we first surveyed the species composition of macrophytes affixed to worm tube caps at three sites in coastal Virginia, USA using transect and quadrat sampling. These unmanipulated field surveys supported previous work revealing variable, but often high frequencies (31-98%) of D. cuprea decoration with G. vermiculophylla. We next used field manipulations and controlled laboratory experiments to test the consistency of individual D. cuprea decoration with G. vermiculophylla versus three common macrophytes (Ulva sp., Agardhiella sp., and Spartina alterniflora) found in our field surveys. Twenty-four hours after removing the worm's tube cap in the field, D. cuprea decoration was dominated by both G. vermiculophylla (39.6%) and S. alterniflora (25.9%). When provided a choice of macrophytes in the laboratory, individual D. cuprea consistently decorated with G. vermiculophylla (58.7%) over the other macrophytes, showing a preference for the non-native macrophyte. Our study suggests that preference can drive strong and steadfast interactions between native and non-native organisms, facilitating the latter's persistence and spread, change available habitat, and alter community interactions.

Invasion of the body snatchers: the role of parasite introduction in host distribution and response to salinity in invaded estuaries.

In dynamic systems, organisms are faced with variable selective forces that may impose trade-offs. In estuaries, salinity is a strong driver of organismal diversity, while parasites shape species distributions and demography. We tested for trade-offs between low-salinity stress and parasitism in an invasive castrating parasite and its mud crab host along salinity gradients of two North Carolina rivers. We performed field surveys every six to eight weeks over 3 years to determine factors influencing parasite prevalence, host abundance, and associated taxa diversity. We also looked for signatures of low-salinity stress in the host by examining its response (time-to-right and gene expression) to salinity. We found salinity and temperature significantly affected parasite prevalence, with low-salinity sites (less than 10 practical salinity units (PSU)) lacking infection, and populations in moderate salinities at warmer temperatures reaching prevalence as high as 60%. Host abundance was negatively associated with parasite prevalence. Host gene expression was plastic to acclimation salinity, but several osmoregulatory and immune-related genes demonstrated source-dependent salinity response. We identified a genetic marker that was strongly associated with salinity against a backdrop of no neutral genetic structure, suggesting possible selection on standing variation. Our study illuminates how selective trade-offs in naturally dynamic systems may shape host evolutionary ecology.

Invasion history shapes host transcriptomic response to a body-snatching parasite.

By shuffling biogeographical distributions, biological invasions can both disrupt long-standing associations between hosts and parasites and establish new ones. This creates natural experiments with which to study the ecology and evolution of host-parasite interactions. In estuaries of the Gulf of Mexico, the white-fingered mud crab (Rhithropanopeus harrisii) is infected by a native parasitic barnacle, Loxothylacus panopaei (Rhizocephala), which manipulates host physiology and behaviour. In the 1960s, L. panopaei was introduced to the Chesapeake Bay and has since expanded along the southeastern Atlantic coast, while host populations in the northeast have so far been spared. We use this system to test the host's transcriptomic response to parasitic infection and investigate how this response varies with the parasite's invasion history, comparing populations representing (i) long-term sympatry between host and parasite, (ii) new associations where the parasite has invaded during the last 60 years and (iii) naïve hosts without prior exposure. A comparison of parasitized and control crabs revealed a core response, with widespread downregulation of transcripts involved in immunity and moulting. The transcriptional response differed between hosts from the parasite's native range and where it is absent, consistent with previous observations of increased susceptibility in populations lacking exposure to the parasite. Crabs from the parasite's introduced range, where prevalence is highest, displayed the most dissimilar response, possibly reflecting immune priming. These results provide molecular evidence for parasitic manipulation of host phenotype and the role of gene regulation in mediating host-parasite interactions.

Physiological impacts of time in holding ponds, biomedical bleeding, and recovery on the Atlantic horseshoe crab, Limulus polyphemus.

Atlantic horseshoe crabs, Limulus polyphemus (HSC), are commercially harvested along the eastern U.S. coast and bled for hemolymph used in pharmaceutical safety testing. In South Carolina, some HSCs are held in outdoor ponds before transport to facilities where they are bled and then released to the wild. This study examined whether the time HSCs are held before bleeding, bleeding itself, or the duration of the recovery after bleeding affects HSC mortality and physiological condition. Female HSCs were collected from Coffin Point Beach, South Carolina (April 22-24, 2016), held in ponds for 2, 4, 6, or 8 weeks, then bled or held as controls. Body weights, hemocyanin concentrations, and hemocyte densities were measured prior to treatment (bled/control) and at 2, 6, and 12 days of recovery. Hemocyanin concentrations declined significantly in HSCs held in ponds for 8 weeks prior to bleeding and were excluded from further analyses. Compared to some studies, HSC mortalities were low (11%). Impacts of time in holding ponds, bleeding, and recovery from bleeding on physiological measures were assessed using 3-way fixed-effects ANOVA. While duration of recovery had main effects on physiological measures, significant interactions were also present. There was an interaction of treatment and recovery duration, with control crabs having higher hemocyte densities than bled animals at days 2 and 6 of recovery. There were two significant two-way interactions influencing hemocyanin concentration: pond time and recovery, and treatment and recovery. Our study suggests both main and synergistic effects are important when assessing the physiology and mortality of HSCs harvested for biomedical purposes.

Incorporating Ploidy Diversity into Ecological and Community Genetics.

Studies in ecological and community genetics have advanced our understanding of the role of intraspecific diversity in structuring communities and ecosystems. However, in near-shore marine communities, these studies have mostly been restricted to seagrasses, marsh plants, and oysters. Yet, macroalgae are critically important ecosystem engineers in these communities. Greater intraspecific diversity in a macroalgal ecosystem engineer should result in higher primary and secondary production and community resilience. The paucity of studies investigating the consequences of macroalgal intraspecific genetic variation might be due, in part, to the complexity of macroalgal life cycles. The majority of macroalgae have seemingly subtle, but in actuality, profoundly different life cycles than the more typical animal and angiosperm models. Here, we develop a novel genetic diversity metric, PHD , that incorporates the ratio of gametophytic to sporophytic thalli in natural populations. This metric scales from 0 to 1 like many common genetic diversity metrics, such as genotypic richness, enabling comparisons among metrics. We discuss PHD and examples from the literature, with specific reference to the widespread, red seaweed Agarophyton vermiculophyllum. We also discuss a sex diversity metric, PFM , which also scales from 0 to 1, but fewer studies have identified males and females in natural populations. Nevertheless, by incorporating these novel metrics into the repertoire of diversity metrics, we can explore the role of genetic diversity in community and ecosystem dynamics with an emphasis on the unique biology of many macroalgae, as well as other haplodiplontic taxa such as ferns, foraminiferans, and some fungi.

Passive biomonitoring for per- and polyfluoroalkyl substances using invasive clams, C. fluminea.

Per- and polyfluoroalkyl substances (PFAS) are a class of toxic manufactured chemicals in commercial and consumer products. They are resistant to environmental degradation and mobile in soil, air, and water. This study used the introduced bivalve Corbicula fluminea as a passive biomonitor at sampling locations in a primary drinking water source in Virginia, USA. Many potential PFAS sources were identified in the region. Perfluorohexane sulfonate (PFHxS) and 6:2 fluorotelomer sulfonic acid (6:2 FTS) levels were highest downstream of an airport. The highest levels of short-chain carboxylic acids were in locations downstream of a wastewater treatment plant. Measured PFAS concentrations varied by location in C. fluminea, sediment, and surface water samples. Two compounds were detected across all three mediums. Calculated partitioning coefficients confirm bioaccumulation of PFAS in C. fluminea and sorption to sediment. C. fluminea bioaccumulated two PFAS not found in the other mediums. Perfluoroalkyl carboxylic acids and short-chain compounds dominated in clam tissue, which contrasts with findings of accumulation of longer-chain and perfluorosulfonic acids in fish. These findings suggest the potential for using bivalves to complement other organisms to better understand the bioaccumulation of PFAS and their fate and transport in a freshwater ecosystem.

Marine invasions and parasite escape: updates and new perspectives.

Marine invasions have risen over time with enhanced globalization, and so has the introduction of non-native hosts and their parasites. An important and well-supported paradigm of invasion biology is the significant loss of parasites that hosts enjoy in introduced regions compared to native regions (i.e. parasite escape), yet less is known about the factors that influence parasite escape in marine systems. Here, we compile an up-to-date review of marine parasite invasions and test several hypotheses related to host invasion pathway that we suspected could influence parasite escape across the 31 host-parasite systems included in our investigation. In general, we continued to show significant support for parasite escape; however, escape varied among parasite taxa, with most taxa demonstrating moderate levels of escape and a few showing complete or no escape. Moreover, we revealed several important factors related to host taxa, geography, time, and vector of introduction that influenced parasite escape, and in some cases demonstrated significant interactions, revealing the complexity of the invasion pathway in filtering parasites from native to introduced regions. In some (but not all) cases, there was also evidence of invasive host advantages due to parasite escape, but more evidence is required to demonstrate clear support for the enemy release hypothesis. In general, our study revealed the need for further research across systems, especially in understudied regions of the world.

Recent introductions reveal differential susceptibility to parasitism across an evolutionary mosaic.

Parasitism can represent a potent agent of selection, and introduced parasites have the potential to substantially alter their new hosts' ecology and evolution. While significant impacts have been reported for parasites that switch to new host species, the effects of macroparasite introduction into naïve populations of host species with which they have evolved remain poorly understood. Here, we investigate how the estuarine white-fingered mud crab (Rhithropanopeus harrisii) has adapted to parasitism by an introduced rhizocephalan parasite (Loxothylacus panopaei) that castrates its host. While the host crab is native to much of the East and Gulf Coasts of North America, its parasite is native only to the southern end of this range. Fifty years ago, the parasite invaded the mid-Atlantic, gradually expanding through previously naïve host populations. Thus, different populations of the same host species have experienced different degrees of historical interaction (and thus potential evolutionary response time) with the parasite: long term, short term, and naïve. In nine estuaries across this range, we examined whether and how parasite prevalence and host susceptibility to parasitism differs depending on the length of the host's history with the parasite. In field surveys, we found that the parasite was significantly more prevalent in its introduced range (i.e., short-term interaction) than in its native range (long-term interaction), a result that was also supported by a meta-analysis of prevalence data covering the 50 years since its introduction. In controlled laboratory experiments, host susceptibility to parasitism was significantly higher in naïve hosts than in hosts from the parasite's native range, suggesting that host resistance to parasitism is under selection. These results suggest that differences in host-parasite historical interaction can alter the consequences of parasite introductions in host populations. As anthropogenically driven range shifts continue, disruptions of host-parasite evolutionary relationships may become an increasingly important driver of ecological and evolutionary change.

Strong genetic structure in a widespread estuarine crab: A test of potential versus realized dispersal.

Aim: Genetic structure has proven difficult to predict for marine and estuarine species with multi-day pelagic larval durations, since many disperse far less than expected based on passive transport models. In such cases, the gap between potential and realized dispersal may result from larval behaviours that evolved to facilitate retention and settlement in favourable environments. Behaviour is predicted to play a particularly key role in structuring truly estuarine species, which often moderate their behaviour to remain within their natal estuaries. In such systems, this restricted dispersal may lead to high divergence, local adaptation and eventual speciation across their range. Here, we test whether a geographically widespread estuarine crab, known to have behaviour promoting larval retention, exhibits high population structure despite a 2- to 4-week larval duration. Location: Atlantic and Gulf Coasts of North America. Taxon: White-fingered mud crab, Rhithropanopeus harrisii. Methods: Population genomic analyses across nine estuaries from New Hampshire to Louisiana using 12,638 transcriptome-derived SNPs. Results: We found highly differentiated genetic signatures among all nine estuaries, separated by 200-5,000 km of coastline. Estimates of gene flow suggest that migration is low and largely symmetrical between sites. We also observed deep phylogenetic divides corresponding to major biogeographical breaks. Main conclusions: These results indicate substantial and longstanding constraints to dispersal in the species' native range, likely arising from the emergence of geological and oceanographic barriers and sustained by behaviour that promotes estuarine retention during larval development. This work supports the idea that larval behaviour promoting estuarine retention can be reflected in substantial genetic structure even in species with multi-week pelagic larval durations. Such behaviour-restricted dispersal has implications for predicting adaptation and spread in estuarine species, many of which have been introduced outside their native ranges.

Assessing the effects of trematode infection on invasive green crabs in eastern north america.

A common signature of marine invasions worldwide is a significant loss of parasites (= parasite escape) in non-native host populations, which may confer a release from some of the harmful effects of parasitism (e.g., castration, energy extraction, immune activation, behavioral manipulation) and possibly enhance the success of non-indigenous species. In eastern North America, the notorious invader Carcinus maenas (European green crab) has escaped more than two-thirds its native parasite load. However, one of its parasites, a trematode (Microphallus similis), can be highly prevalent in the non-native region; yet little is known about its potential impacts. We employed a series of laboratory experiments to determine whether and how M. similis infection intensity influences C. maenas, focusing on physiological assays of body mass index, energy storage, and immune activation, as well as behavioral analyses of foraging, shelter utilization, and conspicuousness. We found little evidence for enduring physiological or behavioral impacts four weeks after experimental infection, with the exception of mussel handling time which positively correlated with cyst intensity. However, we did find evidence for a short-term effect of M. similis infection during early stages of infection (soon after cercarial penetration) via a significant drop in circulating immune cells, and a significant increase in the crabs' righting response time. Considering M. similis is the only common parasite infecting C. maenas in eastern North America, our results for minimal lasting effects of the trematode on the crab's physiology and behavior may help explain the crab's continued prominence as a strong predator and competitor in the region.

'Caribbean Creep' chills out: climate change and marine invasive species.

BACKGROUND: New marine invasions have been recorded in increasing numbers along the world's coasts due in part to the warming of the oceans and the ability of many invasive marine species to tolerate a broader thermal range than native species. Several marine invertebrate species have invaded the U.S. southern and mid-Atlantic coast from the Caribbean and this poleward range expansion has been termed 'Caribbean Creep'. While models have predicted the continued decline of global biodiversity over the next 100 years due to global climate change, few studies have examined the episodic impacts of prolonged cold events that could impact species range expansions. METHODOLOGY/PRINCIPAL FINDINGS: A pronounced cold spell occurred in January 2010 in the U.S. southern and mid-Atlantic coast and resulted in the mortality of several terrestrial and marine species. To experimentally test whether cold-water temperatures may have caused the disappearance of one species of the 'Caribbean Creep' we exposed the non-native crab Petrolisthes armatus to different thermal treatments that mimicked abnormal and severe winter temperatures. Our findings indicate that Petrolisthes armatus cannot tolerate prolonged and extreme cold temperatures (4-6 °C) and suggest that aperiodic cold winters may be a critical 'reset' mechanism that will limit the range expansion of other 'Caribbean Creep' species. CONCLUSIONS/SIGNIFICANCE: We suggest that temperature 'aberrations' such as 'cold snaps' are an important and overlooked part of climate change. These climate fluctuations should be accounted for in future studies and models, particularly with reference to introduced subtropical and tropical species and predictions of both rates of invasion and rates of unidirectional geographic expansion.