Salinity tolerance,salinity preference and temperature tolerance in the high-shore harpacticoid copepod Tigriopus brevicornis

Tigriopus brevicornis (O. F. Müller) were collected in 1992 from rock pools close to U.M.B.S. Millport, Isle of Cumbrae, U.K. and acclimated to various combinations of salinity and temperature for at least 1 wk prior to laboratory experiments. Higher salinities of acclimation enhanced tolerance to high salinity stress, while tolerance of low salinities was hardly affected by acclimation salinity. Acclimation to low temperature (10°C) extended the survivable salinity range for T. brevicornis. High-salinity acclimation enhanced the survivable temperature range. Copepods acclimated to 60 survived significantly lower and higher temperatures than did 34-acclimated individuals. At high temperature, 75-acclimated female copepods had the highest median lethal temperature, 38.9°C. Females were significantly more resistant to high temperatures than males. The copepods were seen to have a very low median lethal temperature when frozen into solid ice for 2 h; 50% mortality occurred at-16.9°C in 10°C, 34-acclimated T. brevicornis. Salinity preference experiments demonstrated an ability to discriminate between salinities differing by as little as 3. Copepods acclimated to 34 chose salinities near their acclimation salinity; individuals acclimated to 5 favoured slightly higher salinities, while copepods acclimated to 60 chose rather lower salinities.     

Identification of sex, age and species-specific proteins on the surface of the harpacticoid copepod Tigriopus japonicus

 Behavioral experiments have shown that male copepods of the species Tigriopus japonicus (Nori) can distinguish species, sex, and developmental stage of potential mates using contact chemoreception. Lectin-binding patterns on the body surface of females have indicated that surface-bound glycoproteins may be important signals in mate choice. In the present study, the proteolytic enzyme trypsin was used to cleave surface proteins from females, reducing their attractiveness to males. The protein fragments released were used to make monoclonal antibodies. Three levels of screening were used to identify monoclonal antibodies that recognized proteins involved in mate recognition. One monoclonal antibody bound to the terminal urosome and lateral prosome of CV females, and its binding significantly decreased female attractiveness to males. Western blotting showed that this antibody bound the trypsin-cleaved fragment and proteins of homogenized CV females and virgin adult females, but did not bind proteins of homogenized males, CIII females, or females of T. californicus or T. fulvus. This antibody recognized proteins on the surface of females that may enable males to discriminate conspecifics, sex, and age. It is likely that this molecule has a central role in the evolution of reproductive isolation in this group. Received: 22 July 1999 / Accepted: 21 March 2000     

Mating system of the intertidal copepod Tigriopus californicus

Male Tigriopus californicus clasp immature females (copepodid stages II–V) for a period of up to a week prior to the female's terminal molt; upon maturation (stage VI) the female is inseminated and released. While females can mate anytime after their terminal molt, experiments using electrophoretically-detected genetic markers indicate that each mates only once in her lifetime. No evidence of sperm displacement was observed. Hence, male mating behavior can be interpreted as pre-copulatory mate guarding, a strategy employed to assure that a potential mate has not been previously inseminated. Males minimize the time investment required to insemiate a single female successfully by preferentially choosing to clasp more developmentally-advanced females; males clasped to stage III females will release them in order to clasp stage V females if the latter are present. Since males are capable of multiple mating, under most conditions of population sex ratio, this mating system results in low availability of unclasped, developmentally-advanced females; consequently, males must clasp successively younger (i.e. developmentally less-advanced) females in order to obtain a successful insemination.     

Role of surface glycoproteins in mate-guarding of the marine harpacticoid Tigriopus japonicus

Surface glycoproteins serve as mate- and gamete-recognition molecules in some marine animals such as rotifers and sea urchins. We investigated the role of contact chemoreception of surface glycoproteins in mate recognition, mate-guarding, and spermatophore transfer in the marine harpacticoid Tigriopus japonicus Mori. Adult males accurately distinguished immature females from a pool of mixed-sex juveniles, and their behavior towards both live and dead females is described. A survey of the binding sites of twelve lectins to the surface of juvenile and adult females, and adult males demonstrated localized lectin-binding at sites considered important in mating behavior of each sex/age class. Treating adult males with the Triticum vulgaris lectin sharply inhibited normal mate recognition and guarding behavior, as did the monosaccharide glucosamine. Treatment of males with protease and detergents destroyed their mate-guarding behavior without affecting swimming behavior. These results suggest that protein receptors on the antennules of the males detect glycoprotein signals on the surface of females, recognizing carbohydrate moieties of glycoproteins to identify appropriate partners for guarding or mating. Surface glycoproteins may function as a common mechanism of sexual communication in many marine zooplankters. Received: 6 May 1997 / Accepted: 14 September 1997     

Tube-building by a marine meiobenthic harpacticoid copepod

Pseudostenhelia wellsi Coull and Fleeger is a meiobenthic harpacticoid copepod inhabiting muddy, estuarine sediments. All individuals observed, and all ages including nauplii, build and inhabit elongate, mucous tubes which may extend to a depth of 3.9 mm into the sediment. Many of the narrow tubes, 0.27 mm in diameter, have a tube-cap extending 0.32 mm above the sediment-water interface. The tubes are a matrix of fine silt, sand and detritus cemented with a mucopolysaccharide, as shown by the Periodic Acid Schiff stain, apparently secreted from glands in the ventrolateral margin of the cephalothorax. Upon addition to sterile, homogeneous sediment, P. wellsi quickly (within 1 to 2 h) transform the upper 0.4 cm to a cohesive conglomerate of tubes, silt-clay particles and mucous filaments.     

Ontogenetic feeding shifts in the meiobenthic harpacticoid copepod Nitocra lacustris

¹⁴C-radiolabelling experiments indicate that adult stages of the salt-marsh harpacticoid copepod Nitocra lacustris (Schmankevitsch) receive a large part of their nutrition through the ingestion and assimilation of certain diatoms. An abundance of empty diatom frustules occurs in the gutpellet contents of field-collected individuals. Naupliar stages do not ingest diatoms in the laboratory, and nauplii from the field do not contain frustules in their gut pellets. Ingestion of diatoms in the laboratory first occurs during the second or third copepodite stage. ³H-radiolabel expeiments and grazing experiments using bacterium-sized beads adhering to the diatoms indicated that both adults and nauplii ingest bacteria adhering to the outer mucus coating of the diatoms (and probably ingest the diatom mucus itself). Adults ingest bacteria (and probably mucus exopolymer) coincidently while ingesting diatoms. The nauplii ingest these components by scraping the outer surface of the diatoms. SEM observations indicate that diatoms are not punctured by the nauplii during feeding. While diatom mucus and associated bacteria play an (as yet unquantified) role in the nutrition of the adults, these components may comprise the bulk of food resources for naupliar stages.     

Environmental stability and seasonality of a harpacticoid copepod community

The southwest monsoon on the west coast of India brings about dynamic changes in estuaries and coastal waters. The response of the meiofauna to monsoonal rain is obvious, but the impact of such environmental changes on the community structure of harpacticoid copepod species and their seasonality in the estuaries influenced by the tropical monsoon is poorly understood. In this study the spatial and temporal variability in abundance and community structure of meiobenthic copepods was investigated over an annual cycle (June 1983 to June 1984), in an estuary influenced by the tropical monsoon. Total meiofaunal abundance showed wide variations in space and time. Minimum and maximum densities were observed in the monsoon and pre-monsoon seasons, respectively. Quick recovery of harpacticoid populations in the early post-monsoon season indicated their recuperative power under adverse conditions. Of the 25 species recorded from lower, middle and upper reaches, eight comprised over 70% of the total copepod population. The peak and low occurrence of dominant species displayed striking correlations with the summer and rainy seasons, respectively. While other species were restricted in distribution, Stenhelia longifurca was recorded from all salinity regimes. The harpacticoid community was greatly influenced by the onset of the monsoon period, and their spatial and temporal variabilities were related with physico-chemical parameters and the variability of these parameters in the estuary.     

Maintenance of a diverse harpacticoid copepod community in microcosm culture

The design and construction of a closed system circulating sea-water tidal estuarine sediment microcosm is described. The microcosm was used to culture mixed species populations of the Harpacticoida (Crustacea: Copepoda) and the evolution of the community and the successional changes are detailed. The communities that developed in each of four replicate microcosms were statistically indistinguishable with respect to the species richness, the numbers of individuals recorded, the log series diversity and the dominance diversity statistics. The most abundant species were the same in each microcosm with the same rank. A diverse community was maintained that did not tend to monoculture; however, the successional changes observed were not apparent in the estuarine communities from which the culture sediments was collected. Rather than a functional reorganisation of the community effecting this, gross environmental changes resulting from a decreased depth of sediment matrix are implicated. The success in maintaining a diverse community is attributed to a provision of a number of nutrient sources.     

Deep-sea harpacticoid copepod diversity maintenance: The role of polychaetes

Several models to account for enhanced diversity in the deep sea have been proposed, but the available natural history information has been inadequate to distinguish among them. In particular, few data exist on patterns of co-occurrence among species. At 1220 m depth in the San Diego Trough (32°35.75N; 117°29.00W), harpacticoid copepod species covary significantly with polychaetes when the polychaetes are combined into functional groups on the basis of feeding type and mobility. In particular, harpacticoid species tend to avoid polychaetes which are sessile surface-deposit feeders. The results provide support for models in which disturbance/predation plays an important role in maintaining deep-sea diversity.Contribution No. 15 from Expedition Quagmire.     

Impact of epibenthic predation on estuarine intertidal harpacticoid copepod populations

The extent to which energy is transferred directly from benthic meiofauna to epibenthic predators was investigated on an intertidal sand-flat in the Exe estuary, southwest England, during 1981–1982 and compared with data obtained from an intertidal mud-flat in the Lyhner estuary, also in south-west England, between 1978 and 1981. Two species of flatfish (Pleuronectes platessa L. and Platichthys flesus L.), two species of goby [Pomatoschistus microps (Krøyer) and P. minutus (Pallas)], brown shrimp (Crangon crangon L.) and shore crabs (Carcinus maenas L.) are the most common epibenthic predators feeding on the benthic invertebrates in these locatites. Harpacticoid copepods are the only component of the meiofauna to form a significant part of the diet of early juvenile stages of these predators, particularly the invertebrates. Harpacticoids are a more important source of food for predators feeding over the sand-flat than for those feeding on the mud-flat because in the sand-flat alternative prey of suitable size, such as small annelids, are absent. Moreover, the impact of predation on the mud-flat is spread over the whole harpacticoid species spectrum whereas on the sandflat it is confined almost entirely to a single species, Asellopsis intermedia (T. Scott). Flatfish, gobies and shrimp consume daily an estimated 0.01 to 0.1% of the standing stock of A. intermedia and account for between 12 and 22% of the observed reduction in the population of this species between July and October. Therefore, only a very small proportion of total meiofauna biomass is transferred directly to higher trophic levels.     

Genetic differentiation and reproductive incompatibility among Baja California populations of the copepod Tigriopus californicus

Populations of the harpacticoid copepod Tigriopus californicus were sampled from five sites from San Diego, California, to Playa Altamira, Baja California, Mexico. Allozyme analyses revealed that all the study populations are sharply differentiated genetically. At the extreme, two populations, Punta Baja and Playa Altamira, have no alleles in common at the seven allozyme loci studied. All pairwise interpopulation crosses successfully produced F₂ hybrids except those involving the Playa Altamira population. All crosses using Playa Altamira females failed to produce F₁ hybrids, while Playa Altamira males successfully produced F₁ progeny with females from all other sites. These F₁ offspring, however, were completely sterile (with San Diego and Punta Banda females) or only occasionally produced F₂ offspring (with Punta Morro and Punta Baja females). These results suggest that allopatric differentiation among Baja populations has resulted in exceptionally high levels of genetic divergence and nearly complete reproductive isolation of the Playa Altamira population, which should now be recognized at the semispecies (or perhaps sibling species) level.     

Mate choice in the face of both inbreeding and outbreeding depression in the intertidal copepod Tigriopus californicus

 In species vulnerable to both inbreeding and outbreeding depression, individuals might be expected to choose mates at intermediate levels of genetic relatedness. Previous work on the intertidal copepod Tigriopus californicus has repeatedly shown that crosses between populations result in either no effect or hybrid vigor in the first generation, and hybrid breakdown in the second generation. Previous work also shows that mating between full siblings results in inbreeding depression. The present study again found inbreeding depression, with full sibling mating causing significant fitness declines in two of the three populations assayed. In the mate choice assays, a single female was combined with two males. Despite the costs of both inbreeding and outbreeding, mate choice showed clear inbreeding avoidance but no clear pattern of outbreeding avoidance. This lack of outbreeding avoidance may be attributed either to the temporary increase in fitness in the F₁ generation or to the absence of selection for premating isolation in wholly allopatric populations with infrequent migration. If this inability to avoid unwise matings is common to other taxa, it may contribute to the problem of outbreeding depression when allopatric populations are mixed together. Received: 18 May 1999 / Accepted: 25 January 2000     

Effects of two paralytic shellfish toxin producing dinoflagellates on the pelagic harpacticoid copepod Euterpina acutifrons

The effects of two paralytic shellfish toxin (PST) producing dinoflagellates, Alexandrium minutum Halim (high and low toxin strains) and Gymnodinium catenatum Graham, on the pelagic harpacticoid copepod Euterpina acutifrons Dana were tested in a series of experiments run from October 1994 to May 1995. In small volumes (350 ml), both strains of A. minutum (300 to 350 cells ml^-1), and G. catenatum (175 cells ml^-1), strongly reduced naupliar activity (about 30 and 17% were inactive after 24 h, respectively). Activity is here defined as movement. In medium volumes (6 litre), 40% of nauplii incubated with the high toxin strain of A. minutum (1000 cells ml^-1) and 8% of nauplii incubated with cell-free filtrate of the same culture were inactive after 24 h; these values increased to 50 and 30% respectively after 3 d. In large volumes (20 litre), adult copepods incubated with A. minutum (1000 and 10000 cells ml^-1) for 5 d revealed only trace levels of PSP-toxins (paralytic shellfish poisoning) in the extracts analysed by HPLC. With both strains of A. minutum (1000 and 10000 cells ml^-1), 10 to 15% of the copepods were inactive after 1 to 2 d. It is suggested that E. acutifrons avoids feeding on the dinoflagellates after tasting a few cells, but that the dinoflagellates may exude toxins or other substances that affect the copepods. The inactivating effect of the toxic dinoflagellates on the nauplii was more rapid and stronger than on adult copepods, although strong inactivation and death were also observed in adults with time (up to 80% were inactive after 5 d of incubation with A. minutum). Still, in our experiments a considerable proportion of adult females incubated with the toxic dinoflagellates remained active and were able to produce viable eggs for several days.     

Feeding deterrent and toxicity effects of apo-fucoxanthinoids and phycotoxins on a marine copepod (Tigriopus californicus)

Using the marine harpacticoid copepod Tigriopus californicus, the effects of phytoplankton feeding deterrents and toxins were differentiated and measured. Eight compounds were tested for feeding deterrence and toxicity responses: four apo-fucoxanthinoids (apo-10′-fucoxanthinal, apo-12′-fucoxanthinal, apo-12-fucoxanthinal, and apo-13′-fucoxanthinone) and four well-known phycotoxins (domoic acid, okadaic acid, microcystin-LR, and a mixture of PSP-1 toxins). Since several of these compounds exhibited both feeding deterrence and toxicity, a model was developed to deconvolute the observed toxicity response from the observed feeding deterrence response, and to classify these compounds based on the degree of toxicity and/or feeding deterrence they exhibited towards T. californicus. Microcystin-LR, the PSP-1 toxins, and the four apo-fucoxanthinoids behaved only as feeding deterrents at low concentrations. Okadaic acid exhibited both toxicity and feeding deterrence at low concentrations, with the threshold concentration for feeding deterrence at a lower level than the threshold concentration for toxicity. Domoic acid acted only as a toxin at low concentrations, with all decreases in feeding resulting from the death of the copepod. Received: 4 November 1996 / Accepted: 3 December 1996     

Evaluation of the copepod Tigriopus californicus as a bioassay organism for the detection of chemical feeding deterrents produced by marine phytoplankton

Marine phytoplankton have been shown to use chemical feeding deterrents to reduce or inhibit zooplankton grazing. In order to screen phytoplankton species for feeding deterrent production and to isolate and identify feeding deterrent compounds, a new, rapid, and reliable laboratory bioassay was developed. This bioassay used the laboratory-reared harpacticoid copepod Tigriopus californicus and measured inhibition of feeding by measuring the fecal pellet production rate. The bioassay was capable of detecting deterrent compounds: (1) adsorbed onto ground fish food (a normally palatable food); (2) dissolved in a mixture of seawater and live Thalassiosira pseudonana cells (a species of diatom which had no feeding deterrent activity); and (3) present in live cell cultures. Method (2) was recommended for use in bioassay-guided fractionation (isolation of chemical compounds), as it was reliable, rapid, accurate, and easy to perform with large numbers of samples. The total bioassay time was < 48 h, and data collection required only a microscope. Methanolic cell extracts of several phytoplankton species were screened for feeding deterrent activity. Extracts from the diatom Phaeodactylum tricornutum and the dinoflagellate Gonyaulax grindleyi gave feeding deterrent responses, while extracts from the diatom Thalassiosira pseudonana gave no feeding deterrent responses. Live P. tricornutum cells deterred feeding at densities of 6x10⁵ cells ml^-1. This bioassay should provide a valuable tool in screening phytoplankton for feeding deterrent compounds and determining the chemical nature of these compounds.     

Phylogeography of the rock-pool copepod Tigriopus brevicornis (Harpacticoida) in the northern North Atlantic, and its relationship to other species of the genus

We investigated relationships among North Atlantic Tigriopus brevicornis populations and their relationships to Mediterranean T. fulvus and North American T. californicus, using crossing experiments and mitochondrial DNA sequencing. All T. brevicornis populations tested were interfertile, while interspecific crosses produced either no offspring or offspring that did not survive past the larval stage, with the exception of a few T. brevicornis × T. californicus crosses that produced mature adults. DNA sequencing of a fragment of mitochondrial cytochrome oxidase I (COI) showed that samples of T. brevicornis from Iceland, the Faroes, Ireland, Scotland and Nova Scotia formed a single shallow clade. In contrast, T. brevicornis from more southern populations in France and Portugal formed a clade with substantially greater branch lengths. Tigriopus brevicornis was monophyletic, and T. brevicornis plus Mediterranean T. fulvus were together also monophyletic. The phylogeography of T. brevicornis closely mirrored that found in T. californicus, with substantially reduced interpopulation divergence at northern latitudes. The known distribution of T. brevicornis in Iceland and the Faroes is shown and dispersal mechanisms and habitat selection briefly discussed.     

Survival and sex ratios of the intertidal copepod,Tigriopus californicus,following ultraviolet-B (290–320 nm) radiation exposure

The marine harpacticoid copepod, Tigriopus californicus (Baker), is a successful colonizer of supralittoral splash pools from Torch Bay, Alaska, to Baja California, Mexico. As these pools are subject to abundant amounts of direct solar radiation, it was of interest to determine the sensitivity of T. californicus to ultraviolet-B (UV-B, 290 to 320 nm) radiation. During 1980, copepods were raised under diurnal conditions in the laboratory and fed a mixture of unicellular algae and bacteria. Larval and adult stages were irradiated for 0 to 26 h on a rotating turntable under enhanced ultraviolet radiation. Following irradiation, the copepods were maintained in the culture area and checked daily for survival. All life stages of T. californicus tolerated enhanced UV-B radiation irrespective of exposure period; in contrast, survival of an irradiated planktonic copepod, Acartia clausii (Giesbrecht), was significantly lower. Additionally, there appeared to be asex ratio shift in adults that developed from the youngest naupliar stages (N₁/N₂) of T. californicus that were irradiated. Extraction and tentative identification of the pigment(s) responsible for the bright orange color of the carapace of T. californicus were performed. These pigments may function to absorb UV radiation and protect internal structures from photochemically induced damage.     

Toxin accumulation and feeding behaviour of the planktonic copepod Calanus finmarchicus exposed to the red-tide dinoflagellate Alexandrium excavatum

The planktonic copepod Calanus finmarchicus is a dominant member of the zooplankton community in the lower St. Lawrence Estuary in eastern Canada. Blooms of the toxic marine dinoflagellate Alexandrium excavatum which produces high cellular levels of paralytic shellfish poisoning (PSP) toxins, occur during the period of high C. finmarchicus production in summer in this region. To study the feeding behaviour of C. finmarchicus in the presence of Alexandrium spp., experiments were conducted in which female adult copepods collected from the St. Lawrence Estuary between May and September 1991 were exposed under controlled conditions to two toxic isolates of A. excavatum (Pr18b and Pr11f) from the estuary and to a non-toxic control (PLY 173) of a closely related species, A. tamarense isolated from the Tamar Estuary, Plymouth, U.K. Clearance rates on non-toxic A. tamarense cells averaged 5.5 ml ind^-1 h^-1 but were nearzero with either toxic isolate. When presented with a mixture of A. excavatum and the non-toxic diatom Thalassiosira weissflogii in varying proportions, C. finmarchicus fed upon the diatom but avoided the toxic dinoflagellate. Although feeding rates on A. excavatum were very low, toxin analysis by high-performance liquid chromatography with fluorescence detection (HPLC-FD) revealed that the PSP toxins were accumulated in copepods exposed to toxigenic dinoflagellates.The toxin composition in copepods was similar to that of the toxic dinoflagellate, but not necessarily identical, particularly after short-term (2-h) exposure, when relatively elevated levels of N-sulfocarbamoyl toxins were detected. The evidence suggests that C. finmarchicus ingests toxic dinoflagellate cells, either mistakenly or during exploratory bouts of feeding, and accumulates PSP toxins in its gut system and perhaps in other tissues.     

What is the sex ratio of harpacticoid copepods in the deep sea?

The sex ratios of deep-sea harpacticoids have been thought to be greatly skewed toward females. The representation of males is notably more equitable (approximately 1:2) at three deep-sea sites (San Diego Trough: 32°52.4N, 117°45.5W, 1 050 m, January 1987; Porcupine Seabight: 51°36.85N, 12°57.30W, 1 369 m, August 1984; northwest Atlantic, 40°27N, 62°20W, 4 820 m, July 1982, June 1983). This ratio conceals extensive variation at the species level. For some species, males are unknown. For other species, males are more numerous than females. Sampling bias against males can occur and may explain the more extreme reports of male rareness in the literature.