Increased macroalgal abundance following mass mortalities of sea urchins (Strongylocentrotus droebachiensis) along the Atlantic coast of Nova Scotia

Summary Recurrent outbreaks of disease between 1980 and 1983 caused catastrophic mortality of sea urchins (>260,000 t fresh weight) along 280 km (straight line distance) of the Atlantic coast of Nova Scotia. The complete elimination of sea urchins and concomitant development of fleshy macroalgal communities have occurred along different parts of this coast in different years. Macroalgal communities in areas where sea urchins died off 1, 3 and 4 years previously are compared to existing sea urchin-dominated barren grounds and to a mature kelp bed without sea urchins. Changes in macroalgal cover and species composition, and increases in biomass, density and size of kelp (Laminaria) species, characterize the succession from barren grounds to 3- and 4-year-old kelp beds. The greatest change occurred between one and three years following sea urchin mass mortality. Within 3 years, kelp beds attained a level of biomass (7.6 kg m^-2) comparable to that of mature beds. Recovery of sea urchin populations via recruitment of planktonic larvae has been slow and spatially variable. Large-scale reciprocal fluctuations in kelp and sea urchin biomass may characterize the trajectory of a dynamic system which cycles between two alternate community states: kelp beds and sea urchin-dominated barren grounds. Periodic decimation of sea urchin populations by disease may be an important mechanism underlying this cyclicity.     

Histopathology of the disease causing mass mortality of sea urchins (Strongylocentrotus droebachiensis) in Nova Scotia

The disease causing mass mortalities of Strongylocentrotus droebachiensis off Nova Scotia, Canada, from 1980 to 1983 is described. Diseased urchins were characterized by loss of preipheral muscle function in tube feet, spines, and mouth. Signs occurred primarily in the body wall and associated tissues (water vascular system, nerves, spine bases) and coelomic fluid. These symptoms were diffuse and included a general infiltration of tissues with amoebocytes. The coelomic fluid often contained reduced numbers of red and white spherule cells, and clotting was incomplete. Progressive breakdown and fragmentation of muscle cells in tube feet and spine bases resulted in destruction of coherent muscle layers and their replacement by numerous spindle-shaped fibrillar muscle remnants. Coelomic lining cells in the tube feet sloughed off into the lumen, but remained in clumps and phagocytosed muscle remnants.     

Egg longevity and time-integrated fertilization in a temperate sea urchin (Strongylocentrotus droebachiensis)

Recent field experiments have suggested that fertilization levels in sea urchins (and other broadcast spawners that release their gametes into the water column) may often be far below 100%. However, past experiments have not considered the potentially positive combined effects of an extended period of egg longevity and the release of gametes in viscous fluids (which reduces dilution rates). In a laboratory experiment, we found that eggs of the sea urchin Strongylocentrotus droebachiensis had high viability for 2 to 3 d. Fertilization levels of eggs held in sperm-permeable egg baskets in the field and exposed to sperm slowly diffusing off a spawning male increased significantly with exposure from 15 min to 3 h. In a field survey of time-integrated fertilizations (over 24, 48, and 72 h) during natural sperm release events, eggs held in baskets accrued fertilizations over as much as 48 h and attained fairly high fertilization levels. Our results suggest that an extended period of egg longevity and the release of gametes in viscous fluids may result in higher natural fertilization levels than currently expected from short-term field experiments.     

Behavioral ecology of Strongylocentrotus droebachiensis (Muller) (Echinodermata: Echinoidea)

Summary Field observation indicated that extensive aggregation behavior occurs in Strongylocentrotus droebachiensis, an important echinoid herbivore in rocky subtidal zones of northern New England. The relationship of this behavior to the behavioral ecology of this species was investigated. Laboratory studies indicated that urchins are chemically sensitive to the stimuli of food and other urchins. Quantitative field experiments with tagged animals and transects showed the existence of two types of aggregations: feeding and nonfeeding, which appear to be pervasive phenomena in the biology of S. droebachiensis.It is concluded that, depending upon their hunger state, urchins shuttle between feeding aggregations in exposed locations and non-feeding aggregations located in relatively sheltered areas (where they spend most of their time). Individuals of S. droebachiensis aggregate actively through response to other urchins in the form of chemotaxis. The aggregative behavior may be maintained for its selective value in terms of defense from predation and, to lesser extents, facilitation of feeding and breeding.     

Polyfluorochrome marking slows growth only during the marking month in the green sea urchin Strongylocentrotus droebachiensis

Abstract. Used singly, the fluorochrome tags tetracycline and calcein have yielded important insights into sea urchin biology, especially regarding growth. We present a new method of tagging using sequential fluorochrome markers, as well as a more precise method of quantifying growth. Such polyfluorochromes enable repeated markings that allow measurement of multiple growth points and unique identification of individuals or groups. We marked sea urchins, Strongylocentrotus droebachiensis , with four fluorochromes: alizarin complexone, calcein, calcein blue, and tetracycline. All fluorochromes marked both by injection and immersion. We examined the clarity of the mark produced with low, metabolically scaled doses, and higher doses similar to those that have been previously used. We tested the effect of fluorochromes on survival, growth, jaw size, and gonad size by marking a size range (3.9–44.3 mm in diameter) of urchins with either one or all four fluorochromes. We quantified growth using a nominal diameter, that is a fitted constant, times the cube root of weight, which increased the precision of measurements by a factor of six relative to measured diameter. Growth rate was a decreasing function of diameter except for a growth lag in the smallest urchins. Growth rate data for all sizes were fitted using: gamma distributions; Tanaka functions; and, for larger sizes, straight lines (von Bertalanffy model). All treatments produced clear marks, with higher doses producing more reliably clear marks. Tetracycline marking did not affect growth; other treatments produced only transient slowing of growth in the marking month. Growth rate, survival, gonad production, and jaw weight did not differ between control and treatments during the following 6 months. Thus, polyfluorochromes produce reliable marks that do not significantly affect growth or gonad production.     

Paramoeba sp. (Amoebida, Paramoebidae) as the possible causative agent of sea urchin mass mortality in Nova Scotia

An amoeba resembling Paramoeba has been cultured from tissues of diseased sea urchins. Cultures containing the amoeba produced signs of the disease when injected into the coelom of healthy urchins. Control treatments lacking the amoeba did not cause the disease. The amoeba was cultured from radial nerve fragments and seen in tissue sections from experimentally infected urchins. Cultures of the amoeba from these experimentally infected urchins reproduced the disease in healthy urchins by both injection and water-borne routes. These observations suggest involvement of Paramoeba in recent mass mortalities of sea urchins in Nova Scotia.     

A morphological analysis of the first cleavage mitotic cytoskeleton isolated from sea urchin embryos (Strongylocentrotus droebachiensis)

Morphological changes in the mitotic cytoskeleton (MC) that occurred through the course of the first cleavage of Strongylocentrotus droebachiensis, cultured at 8°C or at 0°C, temperatures within the natural range for this species, have been investigated. Electron microscopy of MCs isolated from zygotes has revealed that they consisted largely of microtubules (mts). Thus, the morphology of these MCs is derived from the arrangement of the mts which form them. During anaphase, astral rays elongated while kinetochore fibers shortened. Asters enlarged during anaphase as a result of two events: astral ray lengthening and centrosphere enlargement. At the end of anaphase, asters of 8°C MCs filled the entire cell volume. The pattern of changes that occurred in 8°C mitotic apparatuses (MAs) also occurred in 0°C MCs. The observation of asters in 0°C MCs is contradictory to that of Stephens ('72b), who reported that 0°C MCs in this species were anastral. However, in 0°C metaphase MCs, the astral rays and spindle fibers were not as long as those in 8°C MCs. Also in 0°C MCs, the centrosphere was largely filled with dense material, whereas the centrosphere in 8°C MCs was larger and contained little dense material. Asters of 0°C MCs did not attain a large enough size to fill the egg volume completely, as did asters of 8°C MCs.     

Larval settlement of the green sea urchin, Strongylocentrotus droebachiensis, in the southern Gulf of Maine

Abstract. Settlement timing and differential settlement for the larval stage of the green sea urchin, Strongylocentrotus droebachiensis , in the southern Gulf of Maine was studied during the summer of 1996. Settlement densities on astroturf panels were highest in June and early July (13 to 37 m ⁻² d⁻¹), and peaked in mid-June (199 m⁻² d⁻¹). Settlement was low to nonexistent from mid-July through August (0 to 2 m⁻² d⁻¹). During the peak in settlement, no selection for substrate type was observed. In the remainder of the settlement period, differential settlement occurred, with a preference for substrate covered with live coralline algae. Test diameter of newly settled urchins varied among the substrates, with urchins settling on live coralline algae having the largest test diameter (0.43 ± 0.01 mm). There were no differences in test diameter among the different weeks in which sampling was done. Sustained onshore winds occurred only during peak settlement, suggesting that wind drift currents may concentrate larvae and influence patterns of larval settlement.     

Dynein-like Mg2+-ATPase in mitotic spindles isolated from sea urchin embryos (Strongylocentrotus droebachiensis)

Two distinctly different ATPases have been reported to be endogenous to the mitotic apparatus: a Mg2+-ATPase resembling axonemal dynein, and a Ca2+-ATPase postulated to be bound in membranes. To examine the nature of the Mg2+-ATPase, we isolated membrane-free mitotic spindles from Stronglylocentrotus droebachiensis embryos by rapidly lysing these in a calcium-chelating, low-ionic-strength buffer (5 mM EGTA, 0.5 mM MgCl2, 10 mM PIPES, pH 6.8) that contained 1% Nonidet P-40. The fibrous isolated mitotic spindles closely resembled spindles in living cells, both in general morphology and in birefringence. In electron micrographs, the spindles were composed primarily of microtubules, free from membranes and highly extracted of intermicrotubular cytoplasmic ground substance. As analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), the pelleted spindles contain 18% tubulin, variable amounts of actin (2-8%), and an unidentified protein of 55 kdaltons in a constant weight ratio to tubulin (1:2.5). The isolated spindles also contained two polypeptides, larger than 300 kdaltons, that comigrated with egg dynein polypeptides, and ATPase activity (0.02 mumol Pi/mg . min) that closely resembled both flagellar and egg dynein. The spindle Mg2+-ATPase showed a ratio of Ca2+-/Mg2+-ATPase = 0.85, had minimal activity in KCl and EDTA, and cleaved GTP at 35% of the rate of ATP. The Mg2+-ATPase was insensitive to ouabain or oligomycin. The spindle Mg2+-ATPase was inhibited by sodium vanadate but, like egg dynein, was less sensitive to vanadate than flagellar dynein. The spindle Mg2+- ATPase does not resemble the mitotic Ca2+-ATPase described by others. We propose that the spindle Mg2+-ATPase is egg dynein. Bound carbohydrate on the two high-molecular-weight polypeptides of both egg dynein and the spindle enzyme suggest that these proteins may normally associate with membranes in the living cell.     

Sperm polymorphism within the sea urchin Strongylocentrotus droebachiensis: divergence between Pacific and Atlantic oceans

The rapid evolution of traits related to fertilization such as sperm morphology may be pivotal in the evolution of reproductive barriers and speciation. The sea urchin Strongylocentrotus droebachiensis has a circumarctic distribution and shows substantial genetic subdivision between northeastern Atlantic populations and northwestern Atlantic and Pacific populations. Using transmission electron microscopy, we show here that sperm shape, size, and ultrastructure differ markedly among populations of S. droebachiensis from different oceans and reflect patterns of genetic divergence. Sperm nuclei from northwestern Atlantic and Pacific populations were longer and narrower than those from the northeastern Atlantic. We additionally demonstrate population-level differences in the amount and location of filamentous actin (F-actin) prior to the occurrence of the acrosome reaction. Sperm from Pacific and northwest Atlantic populations differed from that of all other echinoids examined in that intact sperm contains a partly preformed acrosomal process, a structure more closely resembling the acrosomal rod seen in some molluscs. Immunofluorescent studies using anti-bindin antibodies and the F-actin-specific stain phalloidin confirmed these findings. Divergence of reproductive traits such as sperm morphology may be related to divergence in gamete compatibility and genetic divergence, and could represent the first stages of speciation in free-spawning marine invertebrates.     

Echinoids,epizootics and ecological stability in the rocky subtidal off Nova Scotia,Canada

Mass mortalities ofStrongylocentrotus droebachiensis, attributed to disease, have occurred along the Atlantic coast of Nova Scotia from 1980 to 1982. An amoeboid protist has been tentatively identified as the pathogenic agent. Temperature appears to play an important role in triggering epizootics and determining the extent of mortality. Epizootics have occurred in the autumn of each year during months of record high temperatures. The virtual elimination of echinoids over a large area of coast has enabled regeneration of kelp beds. Disease may play a key role in determining the structure and stability of benthic communities off Nova Scotia and elsewhere by controlling echinoid abundance.     

Colonization, dispersal, and hybridization influence phylogeography of North Atlantic sea urchins (Strongylocentrotus droebachiensis)

We used frequency-based and coalescent-based phylogeographic analysis of sea urchin (Strongylocentrotus droebachiensis) mitochondrial DNA (mtDNA) sequences and previously published microsatellite data to understand the relative influence of colonization and gene flow from older (north Pacific) and younger (northeast Atlantic) sea urchin populations on genetic variation in the northwest Atlantic. We found strong evidence of survival of northwestern Atlantic populations in local Pleistocene glacial refugia: most haplotypes were the same as or closely related to Pacific haplotypes, with deep gene genealogies that reflect divergence times within the northwestern Atlantic that are much older than the last glacial maximum. We detected gene flow across the North Atlantic in the form of haplotypes shared with or recently descended from European populations. We also found evidence of significant introgression of haplotypes from a closely related species (S. pallidus). The relative magnitude of gene flow estimated by coalescent methods (and the effective population size differences among oceanic regions) depended on the genetic marker used. In general, we found very small effective population size in the northeastern Atlantic and high trans-Arctic gene flow between the Pacific and northwestern Atlantic. Both analyses suggested significant back-migration to the Pacific. However, microsatellites more strongly reflected older Pacific migration (with similar effective population sizes across the Arctic), whereas mtDNA sequences appeared to be more sensitive to recent trans- Atlantic dispersal (with larger differences in effective population size). These differences across marker types might have several biological or methodological causes, and they suggest caution in interpretation of the results from a single locus or class of markers.     

Calcium uptake and release by isolated cortices and microsomes from the unfertilized egg of the sea urchin Strongylocentrotus droebachiensis

Isolated cortices from unfertilized sea urchin eggs sequester calcium in an ATP-dependent manner when incubated in a medium containing free calcium levels characteristic of the resting cell (approximately 0.1 microM). This ATP-dependent calcium uptake activity was measured in the presence of 5 mM Na azide to prevent mitochondrial accumulation, was increased by oxalate, and was blocked by 150 microM quercetin and 50 microM vanadate (known inhibitors of calcium uptake into the sarcoplasmic reticulum). Cortical regions preloaded with 45Ca in the presence of ATP were shown to dramatically increase their rate of calcium efflux upon the addition of (a) the calcium ionophore A23187 (10 microM), (b) trifluoperazine (200 microM), (c) concentrations of free calcium that activated cortical granule exocytosis, and (d) the calcium mobilizing agent inositol trisphosphate. This pool of calcium is most likely sequestered in the portion of the egg's endoplasmic reticulum that remains associated with the cortical region during its isolation. We have developed a method for obtaining a high yield of purified microsomal vesicles from whole eggs. This preparation also demonstrates ATP-dependent calcium sequestering activity which increases in the presence of oxalate and has similar sensitivities to calcium transport inhibitors; however, the isolated microsomal vesicles did not show any detectable release of calcium when exposed to inositol trisphosphate.     

Stereological analysis of nutritive phagocytes and gametogenic cells during the annual reproductive cycle of the green sea urchin, Strongylocentrotus droebachiensis

Abstract. The germinal epithelium of sea urchin gonads contains two interdependent populations of cells: somatic cells called nutritive phagocytes (NPs) and germ cells—ovarian and testicular gonial cells and their derivatives, gametocytes and gametes. Annually, NPs vary their structure and function to produce a changing microenvironment for germ cells during gametogenesis and after spawning. Here, we describe seasonal changes in the NPs as they interact with germ cells during successive gametogenic stages in both sexes of the green sea urchin, Strongylocentrotus droebachiensis. Monthly samples were collected from the Gulf of Maine at a depth of 10 m and analyzed with light microscopy and stereology. Shorter day length and falling seawater temperatures were correlated with nutrient mobilization from NPs, initiation of gonial cell mitosis, and subsequent gametogenesis. During gametogenesis, NPs in both sexes were depleted of nutrients and eventually phagocytize residual ova or spermatozoa. Observations from this study are important for understanding both the cellular aspects of the reproductive biology of sea urchins and those environmental factors that affect the onset and progress of gametogenesis.     

Nutritive phagocyte incubation chambers provide a structural and nutritive microenvironment for germ cells of Strongylocentrotus droebachiensis, the green sea urchin

Here we characterize the germinal epithelia of both sexes of Strongylocentrotus droebachiensis, the green sea urchin, throughout its annual gametogenic cycle, using light and electron microscopy and cytochemistry. In both sexes, germinal epithelia include two interacting cellular populations: nutritive phagocytes (NPs) and germ cells. After spring spawning, NPs accumulate nutrients; amitotic oogonia and often mitotic spermatogonia occur in clusters beneath NPs; and subsequent gametogenic stages are residual or absent. During the summer, NP nutrients are mobilized for use in vitellogenesis by residual primary oocytes or to support limited spermatogenesis. In addition, some residual primary oocytes may degenerate and be phagocytized by NPs. Significant nutrient mobilization from NPs and substantial gonial cell mitoses (indicative of new gametogenesis) occur in the fall. In both sexes, all of these changes are facilitated by NPs that form basal incubation chambers near the gonadal wall and within which germ cells are surrounded by nutrients released from the NPs. In females, germ cells at several stages of gametogenesis may be housed in separate chambers in the same NP. Primary oocytes also carry out jelly coat formation, meiosis, and cortical granule translocation within NP incubation chambers. In males, many NPs cooperate to provide large continuous chambers that contain spermatogenic cells at diverse stages. In both sexes these chambers persist throughout the year and isolate gametogenesis from the gonadal lumen. NPs become slender and shorten as their nutrients are depleted. Ova or spermatozoa are stored in the gonadal lumen. Post-spawning, NPs phagocytize differentiated germ cells while simultaneously enclosing intact gonial and residual gametogenic cells in basal chambers near the gonadal wall. In light of our observations, we suggest investigating proteins that may be important in the structural, phagocytic, and nutritive functions of NPs and for which corresponding genes have already been identified in the genome of S. purpuratus, the closely related purple sea urchin.     

Characterizing alloparental care in the pilot whale (Globicephala melas) population that summers off Cape Breton,Nova Scotia,Canada

Alloparental care happens when a calf is cared for by an adult that is not their parent. Although alloparental care is common in social mammals, its prevalence is difficult to assess in cetaceans, and has not been studied in Globicephala melas. A population off Cape Breton, Nova Scotia, has been studied from whale‐watching vessels since 1998, during July and August each year. From 2009 to 2011, we collected photo identifications of calves and the adults accompanying them. Alloparental care was considered to be occurring when a calf was identified with more than one companion. We found that 85.7% of calves in 2009, 80.6% of calves in 2010 and 63% of calves in 2011 had alloparents. Mothers were difficult to identify. Nevertheless, none of the other companions of calves were assigned to the same unit as the mother. Five carers were sexed, four of them males. There were no cases of within‐ or between‐year alloparental care reciprocity. It is possible that delayed reciprocity is happening on a larger time scale in this population, but the most likely explanation is that alloparental care is a byproduct of this species’ social structure, with a very small cost to the alloparent's fitness.     

Effects of location,exposure and physical structure on juvenile recruitment of the sea urchin Strongylocentrotus droebachiensis in the Gulf of Maine

Summary

Studies of juvenile recruitment of the green sea urchin Strongylocentrotus droebachiensis in the Gulf of Maine were conducted during the summer of 1995. These experiments confirmed 12 years of previous observations that settlement only occurs during the months of June and July. Settlement panels were placed at a series of sites along the Maine and New Hampshire coastline to compare recruitment in the northeastern and southwestern regions of the Gulf of Maine. The densities of urchins recruiting in Casco Bay and at the Isles of Shoals were two orders of magnitude higher than those from Eastport and Winter Harbor. There was a discontinuity in settlement densities at Penobscot Bay. Experiments conducted at the Isles of Shoals showed a positive relationship between water motion and larval supply, but neither parameter correlated with recruitment density over eight stations. Contrary to previous results, recuitment was greater within natural, as well as, artificial kelp beds compared to urchin barren areas and control panels outside the experimental kelp beds. The impact of changing community structure due to urchin harvesting was discussed as an factor influencing differences in juvenile urchin recruitment.     

American plaice, Hippoglossoides platessoides (Fabricius), predation on green sea urchins, Strongylocentrotus droebachiensis (O. F. Muller), in eastern Newfoundland

Green sea urchins were the most abundant food species (62% by weight) in American plaice (all female) collected from an area in eastern Newfoundland where sandy bottom abuts urchin dominated bedrock. Of secondary importance were capelin (12.9% by weight), which were spawning in the area when the samples were obtained. The results suggest that American plaice may be important predators of urchins in such habitats.