Mating behavior of Labidocera aestiva (Copepoda: Calanoida)

The calanoid copepod Labidocera aestiva Wheeler exhibits a ritualized mating behavior. Sensory hairs and spines and integumental pores on the male and female are believed to play a regulatory role in the mating encounter. Scanning electron microscopy (SEM) revealed approximately 20 unusual pore structures (pit-pores) on the right ventrolateral surface of the female's abdominal segment, the area where the spermatophore coupling device is attached. Prior to spermatophore transfer, the male vigorously strokes this area with the modified endopod of his left fifth leg to prepare the pit-pore region for attachment of a new spermatophore. Transmission electron microscopy (TEM) indicated that the cells underlying each pit-pore may have a secretory function. We hypothesize that the pit-pores secrete a substance which dissolves the cement binding the coupling device to the abdomen and aids the female to remove a discharged spermatophore from her urosome.Contribution No. 116, Harbor Branch Foundation, Inc.     

Mating behavior of Centropages typicus (Copepoda: Calanoida)

Observations were made on approximately 300 mating couples of Centropages typicus Krøyer. Mating behavior consists of 4 distinct stages: pre-copulation, spermatophore extrusion, copulation, and post-copulation. Successful copulation requires proper positioning of the spermatophore complex on the female urosome and is dependent upon precise movements and orientation of the male during the mating encounter. Sub-optimal mating encounters, resulting in the improper placement of the spermatophore complex, were observed and adaptations tending to reduce spermatophore wastage are discussed.Contribution No. 62, Harbor Branch Foundation, Inc.     

Female genital structures of Metridinidae (Copepoda: Calanoida)

The female genital structures of six calanoid copepod species, belonging to the genera Gaussia, Metridia and Pleuromamma, were studied using light and scanning electron microscopy (SEM). The copulatory pores and seminal receptacles are paired in Gaussia and Metridia, but unpaired in Pleuromamma. A thin epicuticle and a spermatophoral plug are the mechanisms by which the pores are closed before and after copulation. The pores open directly into the receptacles, which are reduced to shallow integumentary depressions in Gaussia. The mode of insemination suggests two matings in females of Gaussia and Metridia, but only one in Pleuromamma. Paired gonopores and egg-laying ducts are present in the three genera, with a characteristic, closed semicircular configuration; these are opened during egg-laying by the action of retractor muscles of the gonoporal plates. The seminal ducts, which open into the gonopores, are long, thin and paired in Gaussia and Metridia, whereas in Pleuromamma they are short, broad and unpaired. The opening of the seminal and egg-laying ducts is synchronized. The shell ducts are paired in Gaussia and Metridia, unpaired in Pleuromamma; these arise from glands situated in the lateral expansions of the last prosomite and lead into the distal part of each egg-laying duct in Gaussia and Metridia and of the egg-laying duct receiving the seminal duct in Pleuromamma. The position and anatomy of these structures are compared to those of other families and genera, and a functional interpretation of their morphology is proposed. The main evolutionary trends of the different structural patterns of female calanoid genitalia are presented in tabular form. Received: 30 December 1996 / Accepted: 11 February 1997     

Mating behavior in three species of Pseudodiaptomus (Copepoda: Calanoida)

Quantitative studies of mating behavior between conspecific and heterospecific pairs of three closely related species in the genus Pseudodiaptomus reveal differences in the timing of life-history events, species-specific attraction of mates, distinctive tactile cues, multiple spermatophore attachments, and gametic incomptibility or hybrid inviability. These differences may function as reproductive isolating mechanisms among calanoid copepods. Differences in breeding behaviors and associated morphological features are greater between sympatric than between allopatric species, indicating that selection reinforces both behavioral and morphological divergences.     

Fecundity studies on Acartia tonsa (Copepoda: Calanoida) in standardized culture

Acartia tonsa Dana longevity and egg production data were studied over a 4-year period under standardized culture conditions. Egg-laying patterns and daily rates were evaluated as bioassay tools for measuring the nutritional value of various copepod foods, as well as assessing other aspects of environmental quality. Altogether, 337 females were observed in various aspects of this work. Experiments were performed with newly matured females in loosely covered crystallizing dishes. Fresh water and a standard concentration of algae were supplied daily, and a running daily tally was kept of the eggs produced. The effects of male copepods on egg production were analyzed. Males were either removed after the first eggs were observed or they remained with the female throughout her life span. Daily egg-laying rates of this latter group of females appeared to be more stable, but abnormal embryo development occurred if the males were not removed after fertilization. Likewise, the results of females which were not mated were studied. Life spans were shorter and egg production rates were lower than those of contemporary mated females. Unmated females were subjectively observed to be less active than their mated counterparts. The effects of continuous and temporary starvation on egg-laying were also observed. Under both normal and starved conditions, males appeared to have shorter life spans than either mated or unmated females. During starvation there were no real differences between the life spans or egg productions of mated or unmated females. Non-nutritional particulate matter was evaluated as a food source. It was ingested by the females, but did nothing to prolong the egg-laying period. Finally, the three algal components of the standard ration were evaluated. Females produced equally well with a mixed diet or with Thalassiosira pseudonana (100 m³) or Chroomonas salina (140 m³), the two larger species of this diet, but when Isochrysis galbana (40 m³) was fed, females reacted as if in a starved situation.     

Nutrition studies in the nauplius larva of Calanus pacificus (Copepoda: Calanoida)

Nauplii of Calanus pacificus were raised on a mixture of algae. Details of the mouth-parts, such as denticles, labial palps and lobes, setations and structure of the masticatory teeth were examined by scanning electron microscopy (SEM). Under the experimental conditions (15°C and 300 gC l^-1), exponential growth coefficients for the period Nauplii II–VI were 0.179 for carbon and 0.228 for nitrogen. C:N ratios dropping from 5.1 to 4.7. Growth was isochronous, each stage lasting 1.5 days. Respiratory losses were 15 to 19.6% of body carbon daily. Nauplii raised on a given alga showed higher rates of ingestion in the presence of this food, compared to nauplii switched to other algae. Minimal threshold concentrations for feeding were found, depending on the size of the food offered and ranging from 5.8 gC I^-1 for Lauderia borealis (28.7 m spherical diameter) to 47.1 gC 1^-1 for Chlamydomonas sp. (11.0 m). Unlike the Copepodite I stage, Nauplii II–VI larvae were not able to ingest small cells such as Isochrysis galbana (4.3 m), or very large ones such as Ditylum brightwellii (47.5 m) at more than maintenance rations. Below the critical concentration for maximal feeding, ingestion was clearly dependent on size of the cells offered, but the size-dependent relationship was different for diatoms and non-diatoms. Filtering rates increased from a threshold concentration to a maximal rate at about 50 gC 1^-1, and decreased at higher concentrations. Critical concentrations ranged from 125 gC 1^-1 for L. borealis to 1000 gC 1^-1 for Chlamydomonas sp. Maximal daily rations ranged between 100 and 150% of body carbon.     

Suspected mechano- and chemosensory structures of Temora longicornis (Copepoda: Calanoida)

The antennulary (A1) setae of Temora longicornis Müller are suspected of being mechanosensory. In the present study, the fine structure of the antennule, setae and pegs was examined by scanning and transmission electron microscopy. Two setae arise proximal to each antennule segment junction. The supraaxial (Type 1) seta of each pair is innervated by a pair of ciliary dendrites and has the morphological characteristics of a mechanoreceptor. The sub-axial (Type 2) seta may be chemosensory, and is innervated by a varying number of dendrites, but always more than two. The antennulary pegs do not appear to be innervated. T. longicornis is directionally sensitive to water disturbances, and this is consistent with the physical structure of the setae. Integumental structures on the body and other non-feeding appendages are described. These observations support previous behavioural experiments which suggested the antennule to be the major site for mechanoreception in copepods; however, mechanoreceptors must also occur on other parts of the body.     

Molecular systematics of six Calanus and three Metridia species (Calanoida: Copepoda)

The discrimination of species of the copepod genus, Calanus (Copepoda; Calanoida), is problematical-especially in regions of sympatry. Although the species of Calanus exhibit exceptional morphological similarity, they are quite distinct in genetic character. The DNA base sequences of the mitochondrial large subunit (16S) ribosomal RNA (rRNA) gene unambiguously discriminated C. finmarchicus (Gunnerus 1765), C. glacialis (Jaschnov 1955), C. marshallae (Frost 1974), C. helgolandicus (Claus 1863), C. pacificus (Brodsky 1948), C. sinicus (Brodsky 1965), and C. hyperboreus (Kroyer 1838). Sequence differences among Calanus species for this gene portion range from 7.3% (between C. glacialis and C. marshallae) to 23.9% (between C. glacialis and C. sinicus). Differences among conspecific individuals were approximately 1 to 2%. [These sequence data were determined between April and November 1993; the sequenced domain is similar to that published previously in Bucklin et al. (1992) but are derived from analysis of additional individuals.] Statistical analysis of the sequence data using a variety of tree-building algorithms separated the taxa into one group of species corresponding to the C. finmarchicus group (C. finmarchicus, C. marshallae, and C. glacialis) and another ungrouped set of species corresponding to the C. helgolandicus group (C. helgolandicus, C. pacificus, and C. sinicus). The C. helgolandicus group may be older than the C. finmarchicus group, making the tree topology less reliable in this area. Calanus hyperboreus was an outlier; Nannocalanus minor (Claus 1863) was the outgroup. Similar analysis of Metridia species confirmed that M. lucens (Boeck 1864) and M. pacifica (Brodsky 1948) are distinct species; M. longa (Lubbock 1854) was still more divergent. These sequence data will allow the design of simple, molecular tools for taxonomic identifications. Diagnostic characters, assayed by rapid molecular protocols, will enable biological oceanographers to answer important questions about the distribution and abundance of all life stages (as well as patterns of reproduction) of morphologically similar species, such as those of Calanus.     

Community interactions and zoogeography of the Indian Ocean Candaciidae (Copepoda: Calanoida)

Eighteen species of Candaciidae have been identified from collections made at 339 stations in the Indian Ocean. Most species are zonally distributed; however, on the eastern and western sides of the Indian Ocean, species ranges are extended north or south by boundary currents. Factor analysis was used to cluster phenotypically similar species based on 130 characters taken from the maxilla (a feeding appendage), the first swimming foot, and the fifth foot (a secondary sexual structure). Four morphological clusters were extracted. Clusterings based on separate factor analyses limited to characters from feeding or from sexually adapted appendages are in substantial agreement with the clusters based on composite morphology. Two geographically recurrent groups of species were also identified, one from equatorial waters, one from the central gyre. Each recurrent group is composed of species exceptionally different in body size and belonging to different morphological clusters. Niches of the species are compared using Hutchinson's multidimensional hypervolume as a model. An attempt is made to describe the niche of each species in terms of environmental variables measured at stations where the species was abundant. Environmental variables (dimensions) included in this study are space, time, temperature, food, oxygen and salinity. Mean niche adaptations of most of the species are separable from congeners along at least one niche dimension. It is proposed that community relationships among the Candaciidae developed to prevent both gamete loss and to avoid trophic competition. It is postulated that newly evolving species underwent contemporaneous displacement of both secondary sexual and trophic characters as a condition for sympatry, or diverged in their physiological adaptations to escape sympatric interference.Contribution No. 3686 of the Woods Hole Oceanographic Institution. This research was supported by NSF Grants GB 27405 and GA 43126 and is based on a doctoral dissertation submitted to the Curriculum in Marine Sciences of the University of North Carolina at Chapel Hill.     

Physiological responses ofCalanus finmarchicus andMetridia longa (Copepoda: Calanoida) during the winter-spring transition

The effect of pH on the biological availability of copper to the brine shrimp Artemia franciscana was studied with acclimated and non-acclimated individuals for the pH range 5.5 to 8.5. A chemical speciation model was used to calculate the speciation of copper in a chemically-defined saline solution as a function of pH. The lipid solubility of inorganic copper species was determined in hexadecane/saline and octanol/saline extraction systems. Copper is absorbed across the gut epithelium and accumulation is proportional to time over a 120 min experimental period. The biological availability of copper decreases with a decrease in the pH of the salt solution and a concomitant increase of the cupric ion concentration. Acclimation to the experimental pH has a marked effect on the uptake process, which depends on the buffer used. There is no extraction of copper in hexadecane, but a small amount goes in octanol. There is, however, no direct relation between the accumulation of copper in the shrimps and the extraction of copper in octanol. Multiple regression of the accumulation rates for non-acclimated animals on the calculated copper species concentrations shows that much of the variation in accumulation rate with pH is explained when copper hydroxide and/or copper carbonate species are considered to be the biologically available forms. Alternatively, the observations can be interpreted as the result of competitive binding of protons and copper species for carrier systems.     

Importance of food quality in determining development and survival of Calanus pacificus (Copepoda: Calanoida)

Mixed zooplankton were collected in June and July of 1985 and 1986 from La Jolla Bay, California, USA, and experiments were conducted to determine how selected dinoflagellates affect development and survival of nauplius larvae of Calanus pacificus. We raised nauplii from eggs on nine species of dinoflagellates at concentrations generally >300 g C l^-1, and compared their development and survival to controls reared using the diatom Thalassiosira weissflogii or filtered seawater. Experiments were conducted for 6 d at 17°C. Development and survival rates of the nauplii fell clearly into one of two groups, depending upon the phytoplankton used as food. The first group was characterized by high development rate (0.46 to 0.84 stage d^-1), and by >27% of the original cohort surviving to at least Nauplius IV or V. The five species producing this result were Gymnodinium simplex, G. splendens, Exuviaella marie-lebourae, Gyrodinium dorsum, and T. weissflogii. The second group was characterized by a development rate similar to that in filtered seawater (0.21 to 0.34 stage d^-1), and by nauplii generally failing to molt past the first feeding stage (Nauplius III), often accompanied by high mortality. The five species producing this result were Gyrodinium resplendens, Ptychodiscus brevis, Glenodinium sp., Amphidinium carterae, and Gonyaulax grindleyi. Development rate and survival were not related to cell size or cell carbon, nor to shape or texture (thecate vs athecate dinoflagellates). Poor growth could be related to the absence of some important, but unidentified, nutritional factors. Alternatively, it could be caused by the presence of plant secondary metabolites which are deleterious to growth, a factor we suspect in P. brevis in particular. Prefeeding nauplii exposed to P. brevis lost neuromuscular control prior to becoming lethargic and dying; nutritional deficiencies may not explain these effects. Methods employed in this study provide useful bioassays for detecting chemical interactions between marine plants and animals. Lethal or sublethal effects of dinoflagellates on their most likely potential predators — copepods — may partially explain why they form significant blooms.     

Taxonomic value of minute structures on the genital segment ofPontellina females (Copepoda: Calanoida)

Scanning electron micrographs of the female genital segment ofPontellina spp. reveal fine-scale structures of systematic and phylogenetic interest. The structures are integumental organs and patches of spinules usually overlooked during routine examinations with a light microscope. Number and arrangement of these structures provide a set of character states and delineate a morphocline in accord with phylogenetic relationships hypothesized previously on the basis of other characters. A basic feature of patchiness shown by the spinules is of ontogenetic interest: arrangement of these patches appears to demarcate individual epidermal cells. The adaptive value of spinules and integumental organs is considered in the light of communicative requirements between partners engaged in courtship and copulation. The specimens used were collected on various expeditions in the warm-water belt of the world oceans and mostly studied during 1977.     

Subitaneous and diapause eggs in Mediterranean populations ofPontella mediterranea (Copepoda: Calanoida): A morphological study

Transmission electron microscope studies on the morphology of subitaneous and diapause eggs in the neustonic copepodPontella mediterranea, collected in coastal waters of the Gulf of Naples from May to December 1986, revealed striking structural differences between the two egg types and the existence of a third egg type that was morphologically similar to a diapause egg but which hatched within 2 to 3 d. In smooth, subitaneous eggs the plasma membrane was surrounded by a thin outer chorion as compared to the thick and highly complex four-layer structure enveloping subitaneous eggs with short spines, and diapause eggs. This coat may be assembled just prior to or after deposition since the morphology of developing oocytes was similar in females laying either subitaneous or diapause eggs. The only other major structural difference between the two egg types was the presence of numerous, disc-like bodies in the ooplasm of diapause eggs. Development was arrested for at least 3 mo, prior to the initiation of the first cleavage, suggesting that these eggs were kept dormant by some endogenous and, possibly, structural mechanism preceeding the termination of diapause.Dedicated to B. Scotto di Carlo, teacher and friend, who died tragically at sea during the course of a cruise on December 15, 1988     

Development and expression of sexual maturity in femaleCalanus pacificus (Copepoda: Calanoida) in relation to food quality

Dinoflagellates, which comprise an important part of the phytoplankton in the neritic region of the Southern California Bight, are known to be grazed byCalanus pacificus; rates of ingestion, development and survival of nauplii are influenced by the food quality of dinoflagellates. We have examined the effects of dinoflagellate food quality on reproduction ofC. pacificus females sampled in La Jolly Bay (32°N; 117°W) between 6 April and 4 May 1988. Four sets of experiments were conducted in which copepods were fed five different species:Gymnodinium splendens, Gyrodinium resplendens., G. dorsum, Exuviella mariae lebourae andGymnodinium simplex. These species were selected on the criterion of their success in supporting the growth of nauplii in previous rearing experiments. The experimental culture concentration was maintained at 100µg C 1^–1, near that of the natural seston. As indices of successful reproduction, we measured the percentage of spawning females and the number of eggs laid per female per day. Laboratory results were compared to stages of ovarian maturation of wild females. Four stages of maturation were determined using histological and microscopic observations.Gymnodinium splendens, Exuviella mariae lebourae, andG. simplex yielded clearly inferior results: few females were able to lay eggs repetitively over the first few days of confinement.Gyrodinium resplendens and, above all,G. dorsum yielded the best results. The stage of sexual maturation in situ appears to influence the rhythmicity of egg-laying events. In the presence of high-quality food, females are able to maintain high reproductive rates by sustaining the maturation of successive batches of eggs; good food quality appears to assure this secondary vitellogenesis and the rapid turnover of ovocytes.     

Effects of parental acclimation temperature and egg-incubation temperature on egg-hatching time in Acartia tonsa (Copepoda: Calanoida)

Experimental results indicate that both parental acclimation temperature and egg-incubation temperature have significant effects on the egg-hatching time of Acartia tonsa Dana between 15° and 25°C. Effects of parental acclimation temperature and egg-incubation temperature are additive if the long-term parental acclimation temperature is constant. A. tonsa are sensitive to temperature and changes of temperature during development (in vivo) and after they are laid (in vitro), up to the time of hatching. A temperature change of the parent culture for time periods of 86 h to 8 d is sufficient to change egg-hatching times. The time required for a temperature change to affect egg-hatching times depends on the magnitude and direction of the temperature change. Field collected A. tonsa demonstrate that changes in egg-hatching times occur in the field populations and can be predicted by mean weekly water temperature.     

Relationship between mouthpart structures and in situ feeding habits of species of the family Pontellidae (Copepoda: Calanoida)

Pontellid copepods were collected from the surface waters of a tidal front region in the Bungo Channel (the Inland Sea of Japan) in June 1986 to examine the relationship between the morphology of cephalic appendages and gut contents. In particular, two dominant species,Labidocera japonica Mori andPontellopsis yamadae Mori, were compared in detail. Large setae on the second maxillae ofL. japonica possessed two rows of setules at right angles to each seta along its inner margin except a terminal part which was serrated, whereas the inner margin of those setae ofP. yamadae was entirely serrated. Judging from the structure of the mouthparts, especially the second maxillae, the former species seems to employ both suspension and raptorial feeding modes, in contrast to the latter, which may use only the raptorial mode of feeding. InP. yamadae, the first maxilla and the maxilliped are also modified for carnivory. Gut content analysis supported the morphological evidence for feeding differences, and revealed thatP. yamadae is a carnivore preying mainly on copepodids whileL. japonica feeds omnivorously on copepod nauplii and phytoplankton particles. Since the mouthpart structures of congeners are quite similar to each other, the feeding behavior and habits might also be similar. Within the family Pontellidae, the generaAnomalocera, Calanopia, Epilabidocera, andPontella have mouthpart structures similar to those ofLabidocera, whereas the genusPontellina resemblesPontellopsis. Morphological similarities would suggest that the first group of genera employs both suspension and raptorial feeding modes, and thatPontellina is a carnivore likePontellopsis.     

Life cycles ofCalanoides acutus,Calanus simillimus andRhincalanus gigas (Copepoda: Calanoida) within the Scotia Sea

The life cycles and distribution of three dominant copepods,Calanoides acutus, Calanus simillimus andRhincalanus gigas were studied from the Discovery collections in the Scotia Sea earlier this century.C. simillimus is a sub-Antarctic species which mates in the top 250 m mainly in spring. The rapid development of the summer generation may allow a second mating period and a smaller second generation to appear in late summer.C. simillimus remains in the surface layers for a longer period thanCalanoides acutus orR. gigas, and its depth distribution is bimodal throughout the winter.R. gigas is most abundant in sub-Antarctic waters to the north of the Polar Front. It mates within the top 750 m later in spring, and development seems less synchronised than that of the other two species, with egg laying and the growth season being more protracted. Stages CIII and CIV are reached by the first autumn and further development resumes very early the following spring. It is not clear whether the majority then spawn or whether a further year may be needed to complete the life cycle. The predominantly Antarctic species,C. acutus mates below 750 m in middle to late winter and the summer generation develops rapidly to either CIV or CV. Its lifespan seems typically 1 yr, but some of the CVs which fail to moult and spawn in winter survive into their second summer, and their subsequent fate is uncertain. The cold-water speciesCalanus propinquus is comparatively rare in the Scotia Sea and aspects of its distribution and life cycle are briefly described for comparison. Regional variations in the timing of these events were apparent forC. simillimus and possiblyCalanoides acutus, but were not seen inR. gigas. Their geographic and vertical separation, together with their asynchronous life cycles support the concept of habitat-partitioning of these dominant herbivores.     

Simple molecular method to distinguish the identity of Calanus species (Copepoda: Calanoida) at any developmental stage

Diagnostic morphological characteristics of copepods of the genus Calanus are restricted largely to minor variations in secondary sex characteristics. This presents a persistent problem in the identification of individuals to species level, especially for immature stages. We have developed a simple molecular technique to distinguish between the North Atlantic Calanus species (C. helgolandicus, C. finmarchicus, C. glacialis and C. hyperboreus) at any life stage. Using the polymerase chain-reaction (PCR), the mitochodrial large subunit (16S) ribosomal RNA (rRNA) gene was amplified from individual copepods preserved in ethanol. Subsequent digestion of the amplified products with the restriction enzymes DdeI and VspI, followed by electrophoretic separation in 2% agarose (Metaphor, FMC Ltd), produced a characteristic pattern for each species. The versatility of the method is demonstrated by the unambiguous identification to species of any life stage, from egg to adult, and of individual body parts. Received: 11 May 1998 / Accepted: 5 August 1998