D. Wayne Coats

Parasites and phytoplankton, with special emphasis on dinoflagellate infections.

Abstract Planktonic members of most algal groups are known to harbor intracellular symbionts, including viruses, bacteria, fungi, and protozoa. Among the dinoflagellates, viral and bacterial associations were recognized a quarter century ago, yet their impact on host populations remains largely unresolved. By contrast, fungal and protozoan infections of dinoflagellates are well documented and generally viewed as playing major roles in host population dynamics. Our understanding of fungal parasites is largely based on studies for freshwater diatoms and dinoflagellates, although fungal infections are known for some marine phytoplankton. In freshwater systems, fungal chytrids have been linked to mass mortalities of host organisms, suppression or retardation of phytoplankton blooms, and selective effects on species composition leading to successional changes in plankton communities. Parasitic dinoflagellates of the genus Amoebophrya and the newly described Perkinsozoa, Parvilucifera infectans, are widely distributed in coastal waters of the world where they commonly infect photosynthetic and heterotrophic dinoflagellates. Recent work indicates that these parasites can have significant impacts on host physiology, behavior, and bloom dynamics. Thus, parasitism needs to be carefully considered in developing concepts about plankton dynamics and the flow of material in marine food webs.

Mixotrophy in gyrodinium galatheanum (DINOPHYCEAE): grazing responses to light intensity and inorganic nutrients*

This paper presents results of field and laboratory studies on mixotrophy in the estuarine dinoflagellate Gyrodinium galatheanum (Braarud) Taylor. We tested the hypotheses that this primarily photosynthetic organism becomes phagotrophic when faced with suboptimal light and/or nutrient environments. In Chesapeake Bay, incidence of feeding of this species on cryptophytes is positively correlated with prey density and concentrations of nitrate and nitrite, but negatively correlated with depth, salinity, and phosphate concentration. Feeding in natural assemblages and cultures increased hyperbolically with light intensity. The stoichiometric proportions of dissolved inorganic P and N (DIP:DIN) at the stations where G. galatheanum was present were far below the optimal growth P:N (1:10). Incidence of feeding was negatively related to the ratio of DIP to DIN, suggesting that P limitation may have induced feeding. Addition of nitrate, or addition of both nitrate and phosphate, inhibited feeding in a natural population, indicating that N limitation may also induce feeding. Ingestion of the cryptophyte, Storeatula major, by cultured G. galatheanum was higher in media low in nitrate or phosphate or both, but moderate rates of feeding occurred in nutrient‐replete cultures. When cells were grown in media with varying concentrations of nitrate and phosphate, N deficiency resulted in greater cellular N and Chl a losses than did P deficiency, but P deficiency stimulated feeding more than N deficiency. Both N and P deficiency, or P:N ratios that deviated greatly from 1:10, result in an increase of cellular carbon content and an increase in propensity to feed. Our results suggest that feeding in G. galatheanum is partly a strategy for supplementing major nutrients (N and P) that are needed for photosynthetic carbon assimilation. Feeding in G. galatheanum may also be a strategy for supplementing C metabolism or acquiring trace organic growth factors, since feeding occurs, although at a reduced rate, in nutrient‐replete cultures.

Spatial and Temporal Aspects of Mixotrophy In Chesapeake Bay Dinoflagellates

ABSTRACT. Gymnodinium sanguineum, Gyrodinium uncatenum, and Ceratium furca are large phototrophic dinoflagellates that commonly form red tides in the mesohaline portion of Chesapeake Bay during the summer. Examination of protargol‐stained specimens revealed that these dinoflagellates also feed heterotrophically as indicated by the presence of food vacuoles containing partially digested prey. Ingested prey were generally identified as nanociliates (≥20 μm) belonging to the oligotrich genera Strobilidium and Strombidium; occasionally other small ciliates (e.g. Balanion sp. and Mesodinium sp.), dinoflagellates, and diatoms were observed in early stages of digestion. the percentage of these mixotrophs that had ingested prey was usually less than 20%, but approached 30% in some samples. Occurrence of food vacuoles in Gymno. sanguineum was positively correlated with ≤20 μm oligotrichous ciliate density; limited data for Gyro. uncatenum suggests a similar relationship, but C. furca feeding was not related to nanociliate densities.

PARASITIC LIFE STYLES OF MARINE DINOFLAGELLATES

Several genera of marine dinoflagellates contain species that have evolved parasitic life styles. Dinoflagellate infections have been reported for a wide range of host organisms including sarcodines. ciliates, free‐living dinoflagellates, various invertebrates, and a few vertebrates. Some dinoflagellates even parasitize other parasitic dinoflagellates. Most species are obligately parasitic and rely on heterotrophy as their sole means of nutrition; however, some are mixotrophic, as they possess chloroplasts during part or all of their life cycle. Many are ectoparasites that use highly specialized structures to attach to their host and feed, while others are intracellular parasites that feed by osmotrophy. Parasitic dinoflagellates often have adverse effects on their host that can lead to reproductive castration or death. The ecological importance of parasitic dinoflagellates is particularly evident during epidemic outbreaks that cause mass mortality of host organisms. Species that infect fish can pose threats to aquaculture. while other species can make commercially important crustacea unpalatable. In the planktonic realm, parasitic dinoflagellates influence the structure and function of the microbial food web. They compete with copepods and other grazers by utilizing ciliates as hosts and can stimulate rapid recycling of nutrients by causing the decline of toxic and non‐toxic red tides.

Parasitism of photosynthetic dinoflagellates by three strains of Amoebophrya (Dinophyta): Parasite survival, infectivity, generation time, and host specificity

Amoebophrya ceratii (Koeppen) Cachon is an obligate parasite of dinoflagellates and may represent a species complex. However, little is known about the biology and host range of different strains of Amoebophrya Cachon. Here, we determined parasite generation time and dinospore infectivity, survival, and ability to infect nonprimary hosts for strains of Amoebophrya from Akashiwo sanguinea (Hirasaka) G. Hansen et Moestrup, Gymnodinium instriatum (Freudenthal et Lee) Coats comb. nov., and Karlodinium micrum (Leadbeater et Dodge) J. Larsen. Akashiwo sanguinea was readily infected, with parasite prevalence reaching 100% in dinospore:host inoculations above a 10:1 ratio. Parasitism also approached 100% in G. instriatum, but only when inoculations exceeded a 40:1 ratio. Karlodinium micrum appeared partially resistant to infection, as parasite prevalence saturated at 92%. Parasite generation time differed markedly among Amoebophrya strains. Survival and infectivity of dinospores decreased over time, with strains from G. instriatum and A. sanguinea unable to initiate infections after 2 and 5 days, respectively. By contrast, dinospores from Amoebophrya parasitizing K. micrum remained infective for up to 11 days. Akashiwo sanguinea and G. instriatum were not infected when exposed to dinospores from nonprimary Amoebophrya strains. Karlodinium micrum, however, was attacked by dinospores of Amoebophrya from the other two host species, but infections failed to reach maturity. Observed differences in host–parasite biology support the hypothesis that Amoebophrya ceratii represents a complex of host‐specific species. Results also suggest that Amoebophrya strains have evolved somewhat divergent survival strategies that may encompass sexuality, heterotrophy during the “free‐living” dinospore stage, and dormancy.

Seasonal abundances of planktonic ciliates and microflagellates in mesohaline Chesapeake Bay waters

Ciliate, heterotrophic microflagellate (hflag) and autotrophic microflagellate (aflag) abundances are reported for mesohaline Chesapeake Bay waters based on samples gathered from April through October 1985–1987. Total water column averages for ciliate and microflagellate abundances were typical of eutrophic marine systems. Ciliate density ranged from 17·2 cells ml−1 in April to 1·8 cells ml−1 in September; hflag ranged from 3·7 × 103 cells ml−1 in June to 1·1 × 103 cells ml−1 in October. In spring the majority of ciliate and hflag standing stocks (70% and 64%, respectively) were located in bottom and transition waters; during summer months the majority (approximately 85% of both groups) were in surface and transition waters. During fall, ciliate stock was concentrated (72%) in surface waters and hflag were relatively evenly distributed in the three water column zones. Ciliate and microflagellate numbers were not directly related to chlorphyll α concentration except in the bottom layer, where simultaneous declines accompanied anoxia. Ciliate concentrations correlated with total numbers of microflagellates and hflag abundance, but not aflag density. We discuss the relative importance of predation and food availability in regulating ciliate and hflag concentrations in mesohaline Chesapeake Bay waters.

Occurrence of the parasitic dinoflagellate Amoebophrya ceratii in Chesapeake Bay populations of Gymnodinium sanguineum

Chesapeake Bay populations of the red‐tide dinoflagellate Gymnodinium sanguineum were regularly infected by the parasitic dinoflagellate Amoebophrya ceratii during the summers of 1988–1991. Infections developed inside the nucleus of G. sanguineum and were always lethal to the host. Parasite generation time was ˜ 40 h at 23° C, with the intracellular, trophont phase lasting 39.5 ± 0.3 h, and the extracellular, vermiform stage persisting for ˜ 20 min. Near surface accumulations of G. sanguineum sometimes exceeded 1,000 cells/ml; however, host abundance was relatively low when integrated over the surface mixed layer of each station (mean = 12.2 cells/ml ± 2.96 SE; n = 60). Parasitized hosts were encountered in 75% of the samples where host abundance was ≥ 1 per ml, and epidemic outbreaks (20–40% hosts infected) were observed on several occasions. Epidemic infections were generally located several meters below surface accumulations of G. sanguineum and were always restricted to a narrow region near the pycnocline. Consequently, integrated station values for parasite prevalence were low, with an average 2.7% (± 0.31 SE; n = 60). Parasite induced mortality removed up to 8% of G. sanguineum populations per day, but averaged < 2% of host biomass throughout the Bay. Thus, parasitism by A. ceratii does not appear to be a major factor regulating G. sanguineum bloom in the main stem of Chesapeake Bay.

Ecology of the Red-Tide Dinoflagellate Ceratium furca: Distribution, Mixotrophy, and Grazing Impact on Ciliate Populations of Chesapeake Bay

Abstract Ceratium furca is a primarily photosynthetic dinoflagellate also capable of ingesting other protists. During 1995 and 1996, we documented the abundance of C. furca in Chesapeake Bay and determined grazing rates on prey labeled with fluorescent microspheres. Abundance usually remained below 20 cells ml−1, although the species was capable of localized late-summer blooms (≤ 478 cells ml−1) in the more saline lower to mid-Bay region. Feeding rates ranged from 0 to 0.11 prey dinoflagellate−1 h−1 or from 0 to 37 pg C dinoflagellate−1 h−1 and were highest at lower salinities. Clearance rates averaged 2.5 ± 0.35 μl dinoflagellate−1 h−1. Impact of C. furca feeding on prey populations was higher in the lower Bay, averaging 67% of Strobilidium spp. removed d−1. Ingestion rates were positively correlated with prey abundance and dissolved inorganic nitrogen, but negatively with salinity, depth, dissolved inorganic phosphorus, and inorganic P:N ratio. Daily consumption of prey biomass by C. furca averaged 4.6% of body carbon, 6.5% of body nitrogen, and 4.0% of body phosphorus, with maximal values of 36, 51, and 32%, respectively. Thus, the ability to exploit an organic nutrient source when inorganic nutrients are limiting may give C. furca a competitive advantage over purely photosynthetic species.

Sexual processes in the life cycle of Gyrodinium uncatenum (Dinophyceae): A morphogenetic overview

Sexual processes in the life cycle of the dinoflagellate Gyrodinium uncatenum Hulburt were investigated in isolated field populations. Morphological and morphogenetic aspects of gamete production, planozygote formation, encystment, excystment, and planomeiocyte division are described from observations of living specimens, Protargol silver impregnated material and scanning electron microscope preparations. The sexual cycle was initiated by gamete formation which involved two asexual divisions of the vegetative organism. Gametes were fully differentiated following the second division and immediately capable of forming pairs. Either isogamous or anisogamous pairs were formed by the mid‐ventral union of gametes. Gametes invariably joined with flagellar bases in close juxtaposition. Complete fusion of gametes required ca. 1 h, involved plasmogamy followed by karyogamy and resulted in a quadriflagellated planozygote. Planozygotes encysted in 24–48 h to yield a hypnozygote capable of overwintering in estuarine sediments. Hypnozygotes collected from sediment in late winter readily excysted upon exposure to temperatures above 15°C. A single quadriflagellated planomeiocyte emerged from the cyst and under culture conditions divided one to two days later. The four flagella were not evenly distributed at the first division and both bi‐ and tri‐flagellated daughter cells were formed.

The Phylogenetic Position of Amoebophrya sp. Infecting Gymnodinium sanguineum

ABSTRACT The small‐subunit rRNA sequence of a species of Amoebophrya infecting Gymnodinium sanguineum in Chesapeake Bay was obtained and compared to the small subunit rRNA sequences of other protists. Phylogenetic trees constructed with the new sequence place Amoebophrya between the remaining dinoflagellates and other protists.