Anthony G. Moss

Bacterial associates of two Caribbean coral species reveal species-specific distribution and geographic variability.

ABSTRACT Scleractinian corals harbor microorganisms that form dynamic associations with the coral host and exhibit substantial genetic and ecological diversity. Microbial associates may provide defense against pathogens and serve as bioindicators of changing environmental conditions. Here we describe the bacterial assemblages associated with two of the most common and phylogenetically divergent reef-building corals in the Caribbean, Montastraea faveolata and Porites astreoides. Contrasting life history strategies and disease susceptibilities indicate potential differences in their microbiota and immune function that may in part drive changes in the composition of coral reef communities. The ribotype structure and diversity of coral-associated bacteria within the surface mucosal layer (SML) of healthy corals were assessed using denaturing gradient gel electrophoresis (DGGE) fingerprinting and 454 bar-coded pyrosequencing. Corals were sampled at disparate Caribbean locations representing various levels of anthropogenic impact. We demonstrate here that M. faveolata and P. astreoides harbor distinct, host-specific bacteria but that specificity varies by species and site. P. astreoides generally hosts a bacterial assemblage of low diversity that is largely dominated by one bacterial genus, Endozoicomonas, within the order Oceanospirillales. The bacterial assemblages associated with M. faveolata are significantly more diverse and exhibit higher specificity at the family level than P. astreoides assemblages. Both corals have more bacterial diversity and higher abundances of disease-related bacteria at sites closer to the mainland than at those furthest away. The most diverse bacterial taxa and highest relative abundance of disease-associated bacteria were seen for corals near St. Thomas, U.S. Virgin Islands (USVI) (2.5 km from shore), and the least diverse taxa and lowest relative abundance were seen for corals near our most pristine site in Belize (20 km from shore). We conclude that the two coral species studied harbor distinct bacterial assemblages within the SML, but the degree to which each species maintains specific microbial associations varies both within each site and across large spatial scales. The taxonomic scale (i.e., phylum versus genus) at which scientists examine coral-microbe associations, in addition to host-elicited factors and environmental fluctuations, must be considered carefully in future studies of the coral holobiont.

Microsatellites reveal origin and genetic diversity of Eurasian invasions by one of the world's most notorious marine invader, Mnemiopsis leidyi (Ctenophora).

Marine invasions are taking place at an increasing rate. When occurring in blooms, zooplanktivorous comb jellies of the genus Mnemiopsis are able to cause pelagic regime shifts in coastal areas and may cause the collapse of commercially important fish populations. Using microsatellites, developed for the first time in the phylum Ctenophora, we show that Mnemiopsis leidyi has colonized Eurasia from two source regions. Our preliminary data set included four sites within the putative source region (US East Coast and Gulf of Mexico) and 10 invaded locations in Eurasian waters. Bayesian clustering and phylogeographic approaches revealed the origin of earlier invasions of the Black and Caspian Sea in the 1980s/1990s within or close to the Gulf of Mexico, while the 2006 invasion of the North and Baltic Seas can be directly traced to New England (pairwise FST = 0). We found no evidence for mixing among both gene pools in the invaded areas. While the genetic diversity (allelic richness) remained similar in the Baltic Sea compared to the source region New England, it was reduced in the North Sea, supporting the view of an initial invasion of Northern Europe to a Baltic Sea port. In Black and Caspian Sea samples, we found a gradual decline in allelic richness compared to the Gulf of Mexico region, supporting a stepping‐stone model of colonization with two sequential genetic founder events. Our data also suggest that current practices of ballast water treatment are insufficient to prevent repeated invasions of gelatinous zooplankton.

Dispersed Oil Disrupts Microbial Pathways in Pelagic Food Webs

Most of the studies of microbial processes in response to the Deepwater Horizon oil spill focused on the deep water plume, and not on the surface communities. The effects of the crude oil and the application of dispersants on the coastal microbial food web in the northern Gulf of Mexico have not been well characterized even though these regions support much of the fisheries production in the Gulf. A mesocosm experiment was carried out to determine how the microbial community off the coast of Alabama may have responded to the influx of surface oil and dispersants. While the addition of glucose or oil alone resulted in an increase in the biomass of ciliates, suggesting transfer of carbon to higher trophic levels was likely; a different effect was seen in the presence of dispersant. The addition of dispersant or dispersed oil resulted in an increase in the biomass of heterotrophic prokaryotes, but a significant inhibition of ciliates, suggesting a reduction in grazing and decrease in transfer of carbon to higher trophic levels. Similar patterns were observed in two separate experiments with different starting nutrient regimes and microbial communities suggesting that the addition of dispersant and dispersed oil to the northern Gulf of Mexico waters in 2010 may have reduced the flow of carbon to higher trophic levels, leading to a decrease in the production of zooplankton and fish on the Alabama shelf.

Protistan epibionts of the ctenophore Mnemiopsis mccradyi Mayer

Mnemiopsis mccradyi, a common coastal ctenophore, was observed to bear two distinct, exclusive assemblages of protistan epibionts. The mobiline peritrich, Trichodina ctenophorii (Estes et al., 1997), and small Flabellula-like gymnamoebae inhabited only the surface of the comb plates. By contrast, small Vexillifera-like gymnamoebae and large Protoodinium-like dinoflagellates were found on the ectoderm. The relationship of the epimicrobial protists with their host varied from possible mutualism (vexilliferids) to commensalism (trichodinids) to parasitism (flabellulids and protoodinids). Trichodinids may benefit from comb plate attachment by enhanced food capture. Although they did not obviously impair comb plate beating, they did distort the surface and appear to produce fissures in the comb plate surface, which could provide inroads for more severe comb plate damage by amoebae. By contrast, when flabellulid amoebae occurred in very high surface densities (up to ∼5000 mm−2), they clearly damaged comb plates by eroding the surface. Where flabellulid pseudopodia invaded the comb plate, we observed local degradation of comb plate cilia, as evidenced by central pair disorientation and plasma membrane perturbation and overt phagocytosis of comb plate cilia. Ectodermal vexilliferids, which occurred at much lower densities, did not appear to have any degradative impact on the ctenophore. By contrast, clusters of ectodermal protoodinids were found in localized depressions most likely caused by invasive phagocytosis. The impact of the protistan assemblages on ctenophore populations is unclear, but under conditions of severe infestation they might depress ctenophore population density.

Metabarcoding reveals environmental factors influencing spatio-temporal variation in pelagic micro-eukaryotes.

Marine environments harbour a vast diversity of micro‐eukaryotic organisms (protists and other small eukaryotes) that play important roles in structuring marine ecosystems. However, micro‐eukaryote diversity is not well understood. Likewise, knowledge is limited regarding micro‐eukaryote spatial and seasonal distribution, especially over long temporal scales. Given the importance of this group for mobilizing energy from lower trophic levels near the base of the food chain to larger organisms, assessing community stability, diversity and resilience is important to understand ecosystem health. Herein, we use a metabarcoding approach to examine pelagic micro‐eukaryote communities over a 2.5‐year time series. Bimonthly surface sampling (July 2009 to December 2011) was conducted at four locations within Mobile Bay (Bay) and along the Alabama continental shelf (Shelf). Alpha‐diversity only showed significant differences in Shelf sites, with the greatest differences observed between summer and winter. Beta‐diversity showed significant differences in community composition in relation to season and the Bay was dominated by diatoms, while the Shelf was characterized by dinoflagellates and copepods. The northern Gulf of Mexico is heavily influenced by the Mobile River Basin, which brings low‐salinity nutrient‐rich water mostly during winter and spring. Community composition was correlated with salinity, temperature and dissolved silicate. However, species interactions (e.g. predation and parasitism) may also contribute to the observed variation, especially on the Shelf, which warrants further exploration. Metabarcoding revealed clear patterns in surface pelagic micro‐eukaryote communities that were consistent over multiple years, demonstrating how these techniques could be greatly beneficial to ecological monitoring and management over temporal scales.

Carbonic Anhydrase and Red Blood Cell Anion Exchange in the Neotenic Aquatic Salamander, Necturus Maculosus

Carbonic anhydrase (CA) activity in blood and other tissues and red blood cell (rbc) anion exchange were measured in the mud puppy, Necturus maculosus, in order to gain insight into the strategy for CO2 transport used by these neotenic salamanders and to further explore evolutionary relationships between rbc CA activity and anion exchange in nonmammalian vertebrates. CA activity was detectable in all of the tissues examined, but CA activity in blood was much lower than that in most vertebrates. There was no indication, however, that additional CA had been incorporated into the membrane fraction of other tissues to compensate for this low blood CA activity. In further contrast to most other animals, low levels of CA activity were also detectable in mud puppy plasma. Preliminary characterization of the rbc CA indicated that the Type II, fast‐turnover enzyme was indeed present, but that there are a very low number of active sites in mud puppy rbcs. Further experiments showed that the rbcs were highly permeable to anions and that the relative rate of anion flux could be inhibited by 4,4‐diisothiocyanostilbene‐2,2‐disulphonic acid. Thus, the process of CO2 transport in the blood of mud puppies probably involves components of the Jacobs‐Stewart cycle, as in most other vertebrates.

Trichodina ctenophorii N. Sp., a Novel Symbiont of Ctenophores of the Northern Coast of the Gulf of Mexico

ABSTRACT. Peritrich ciliates of the genus Trichodina are internal or external symbionts of invertebrate and vertebrate hosts. We describe here Trichodina ctenophorii n. sp., a symbiont of Mnemiopsis mccraydii and Beroë ovata (Phylum Ctenophora). The morphology of fixed and living specimens is revealed by silver impregnation, scanning electron microscopy, and differential interference microscopy. Distinguishing features of Trichodina ctenophorii include a denticular morphology composed of falcate, blunt‐tipped blades, and long, straight thorns, with five pins per denticle. Trichodina ctenophorii is found only on the comb plates of these ctenophores. To the best of our knowledge, this is the first report of a trichodinid from the Gulf of Mexico and the first associated with ctenophores.

Purification and characterization of Salmo gairdneri outer arm dynein

Publisher Summary Dyneins are multimeric ATPases, which make up the inner and outer arms that bridge the outer doublet microtubules of eukaryotic cilia and flagella. They are responsible for the generation of sliding between outer doublets, which in turn is the basis for the formation and propagation of bending waves in both cilia and flagella. Outer arm dyneins are composed of two to three ATPases of M r > 400,000, referred to as the α, β, and, where appropriate, γ heavy chains. Trout sperm is a new source of vertebrate dynein. Sperm can be repeatedly obtained in large quantities from the same trout (up to 4 × 10 spermatozoa per ejaculate), their axonemes can be readily isolated, and the dynein can be extracted efficiently and without significant proteolytic degradation. The advantages of trout sperm have permitted the detailed characterization of trout outer arm dynein to progress rapidly, so that it is now one of the best characterized of all dyneins. This chapter presents an overview of trout physiology and spermatogenesis for those not well acquainted with teleost physiology and anatomy and describes the methods for purification and characterization of Salmo gairdneri outer arm dynein.

Two NAC transcription factors from Citrullus colocynthis, CcNAC1, CcNAC2 implicated in multiple stress responses

NAC (no apical meristem, Arabidopsis transcription activation factor 1 and 2, cup-shaped cotyledon 2) transcription factors (TFs) play important roles in plant growth, development, and responses to abiotic and biotic stress. Two novel NAC TFs were isolated from Citrullus colocynthis, a highly drought-tolerant cucurbit species: CcNAC1 and CcNAC2 each with conserved A–E NAC domains. Subcellular location of CcNAC1 and CcNAC2 investigated via transient expression of 35S::CcNAC1::GFP and 35S::CcNAC2::GFP fusion constructs in Arabidopsis protoplasts, revealed nuclear localization. The transactivation ability of CcNACs was examined in the GAL4 yeast assay system, and showed that only the C-terminal domain of CcNAC1 has the ability to activate reporter genes LacZ and His3. The CcNAC genes accumulated in a tissue-specific manner with expression levels in male flowers of C. colocynthis higher than leaves, hypocotyls or roots. Genome walking was used to isolate the CcNAC1 and CcNAC2-promoter regions. A high number of stress-related sequence motifs were detected, especially in the CcNAC1 promoter. C. colocynthis seedlings were treated with PEG, abscisic acid, salicylic acid (SA), jasmonic acid (JA), H2O2, ethylene, gibberellic acid (GA), wounding or salt. High CcNAC1 expression levels were detected following JA application, and wounding, while high CcNAC2 levels followed treatment with GA, JA, SA, and wounding, indicative of differential regulation of these stress responsive TFs in this cucurbit species.

A mechanosensory system that controls feeding in adult Mnemiopsis

We describe here the food groove complex and mechanism of prey capture used by adult Mnemiopsis spp. ctenophores to obtain prey swept into the auricular grooves by feeding currents. Tentilla that emerge from the tentacular groove of the food groove complex extend into the auricular grooves and capture prey upon their sticky surfaces. The prey-laden tentilla contract and drag the prey to the edge of the transport groove, which is also part of the food groove complex. The transport groove undergoes a focal eversion to capture and transport prey orally. Focal eversion exposes the inner ciliated surface of the transport groove as it extends toward the prey. Focal eversion can be evoked by mechanical stimuli from a probe, but only if it is positioned directly over the tentacular groove. We propose that g-cilia located within the tentacular groove are mechanoreceptors whose output triggers a sensory-motor pathway that in turn everts the transport groove. The mechanosensory-motor pathway is ectodermal and sensitive to Mg2+ anesthesia, which defocuses and amplifies eversion. Tentilla are not strictly necessary for eversion to occur, because preparations lacking tentilla can still display eversion; however, they may amplify the sensory signal by interacting with g-cilia as they contract.