Cypris Stage Research Articles

The first record larvae of the genus Peltogaster (Rhizocephala: Peltogastridae) with eyes and unilobed male aesthetasc

The complete larval development of Peltogaster lineata Shiino, 1943 (Rhizocephala: Peltogastridae), including five nauplii and one cypris stage, is described and illustrated using SEM. The development took 3.5–4 days at a water temperature of 22–23 °C. Peltogaster lineata has the peltogastrid type of development. Nauplii possess a large and clearly reticulated flotation collar, six pairs of dorsal shield setae with the U-shaped second pair, long segmented frontolateral horns each with two subterminal setae, and a long seta at the antennal basis. The attachment disc in female cyprids has a flap-like extension at the posterior margin. Cyprids of both sexes possess two sensory setae at the attachment disc. The large male aesthetasc is unilobed, the female subterminal aesthetasc terminates into two thin long filamentous processes. Larvae of P. lineata have distinctly visible nauplius eyes.

Larvae of two parasitic barnacles, Parasacculina pilosella (Van Kampen et Boschma, 1925) (Rhizocephala: Polyascidae) and Sacculina pugettiae Shiino, 1943 (Rhizocephala: Sacculinidae) studied by scanning electron microscopy

The complete larval development of Parasacculina pilosella (Van Kampen et Boschma, 1925) and Sacculina pugettiae Shiino, 1943 including five naupliar stages and one cypris stage is described and illustrated using SEM. P. pilosella and S. pugettiae have a sacculinid type of development. Nauplii possess a naupliar eye, short frontolateral horns with terminal processes, and a ventral process between the furcal rami. Larvae lack a flotation collar, seta 6 on the antennule and a seta on the antennal basis. Cyprids have a nearly straight LO2. Breakage zone and a spinous process are present only in male larvae. Nauplii of the two species differ by the morphology of the furca: in P. pilosella, the furcal rami are longer and not drowned into cuticular sockets. Naupliar antenna of S. pugettiae has a lateral seta on the endopod which is lacking in P. pilosella. Dorsal head shield setae 1 and 2a are present in S. pugettiae nauplii and not found in P. pilosella larvae. In P. pilosella, all dorsal setae have subterminal pores, whereas in S. pugettiae, pores of the setae 2 are shifted proximally. It is possible that the presence/absence of setae 1 and 2a is the distinctive feature of nauplii of the families Sacculinidae and Polyascidae.

The parasitic barnacle Peltogaster reticulata Shiino, 1943 (Rhizocephala,Peltogastridae) from Russian waters of the Sea of Japan: morphological description, molecular identification and complete larval development.

A rhizocephalan parasite of the hermit crab Pagurus minutus Hess, 1865 (Decapoda, Paguridae) is described from Russian waters using morphological and molecular methods. Partial sequences of the mitochondrial cytochrome c oxidase subunit I gene (COI) were used as a genetic marker. We refer this species to Peltogaster reticulata Shiino, 1943 (Rhizocephala, Peltogastridae) based on its morphological description, since genetic data for the species from its type locality are lacking. The species can be distinguished from its known congeners by the reticular pattern of the external cuticle. Peltogaster reticulata is the sister taxon to P. postica Yoshida Osawa, 2011 (in Yoshida et al. 2011), inhabiting the same host in southwestern Japan. The complete larval development of Peltogaster reticulata, including five naupliar and one cypris stage, is described and illustrated using SEM.

A new internal structure of nauplius larvae: A "ghostly" support sling for cypris y left within the exuviae of nauplius y after metamorphosis (Crustacea: Thecostraca: Facetotecta).

Facetotecta, or crustacean "y-larvae," occur in all the world's oceans although the adult forms remain completely unknown. At the metamorphic molt from the last naupliar instar to the terminal cypris larval stage a free carapace, six pairs of natatory thoracopods, and a segmented thorax and abdomen all develop anew. Unlike in earlier molts, the cephalic shield and the so-called "faciotruncal integument" usually remain together at this last naupliar molt, and the posterior "trunk" portion of the exuviae, while hollow, is not empty. In mounted preparations examined by phase contrast or differential interference contrast (DIC) microscopy, a ghost-like image of part of the cypris thorax, particularly the thoracopods and even their setae, is commonly visible inside the naupliar exuviae, and may be universally present in the Facetotecta. To investigate this "ghost," we used DIC and digital photographic stacking, and also scanning electron microscopy, on slide or stub-mounted final naupliar exuviae of an assortment of undescribed species of Facetotecta that had been reared from planktonic lecithotropic nauplii to the cypris stage at Sesoko Island, Okinawa, Japan, and at Keelung and Green Island, Taiwan. These techniques showed that the "ghost" is a delicate, three-dimensional, fibrous structure, essentially a sling-like mold or matrix with struts attached to the outer cuticle and pairs of deep pockets that previously held the thoracopods of the developing cypris y. Whether it is endoskeletal in nature, the (partial) exuvia of an additional instar, remnants of apoptosis, or something else is currently unknown. Nothing similar has been reported in other thecostracans, or in other crustaceans that undergo a similarly abrupt metamorphosis at the last naupliar molt.

The effect of temperature on motility of the nauplius and cypris stages of the acorn barnacle Semibalanus balanoides

The effect of temperature on motility of the nauplius and cypris stages of the acorn barnacle Semibalanus balanoides

Effects of pharmacological compounds on the barnacle larval development, metabolism and settlement

Effects of select pharmacological compounds, with known mode of action in vertebrates, were evaluated on the development, metabolism and settlement of larvae of the common fouling barnacle, Amphibalanus amphitrite. Atrovastatin, a lipid-regulating compound, cetrizine hydrochloride, an anti-histamine, atenolol, a β-blocker, and amlodipine, a calcium-channel blocker were the compounds studied. Nauplii treated with these compounds took more days to reach the cypris stage when compared with the control. These compounds also inhibited the settlement of cyprids on Petri dishes. While exposure to these compounds led to a decrease in the metabolic activity of stage III nauplii, it increased the respiratory rate of cyprids. The results emphasize the role of neurotransmitters and lipids in the development and settlement of barnacle larvae on hard surfaces.

Synchronism of naupliar development of Sacculina carcini Thompson, 1836 (Pancrustacea, Rhizocephala) revealed by precise monitoring

Sacculina carcini is member of a highly-specialized group of parasitic cirripeds (Rhizocephala) that use crabs (Carcinus maenas) as hosts to carry out the reproductive phase of their life cycle. We describe the naupliar development of S. carcini Thompson, 1836 from a very precise monitoring of three different broods from three specimens. Nauplii were sampled every 4 h, from the release of the larvae until the cypris stage. Larval development, from naupliar instar 1 to the cypris stage, lasts 108 h at 18 °C. A rigorous sampling allowed us to describe an additional intermediate naupliar instar, not described previously. Naupliar instars are renumbered from 1 to 5. Nauplius 1 (N1) larvae hatch in the interna; N2 are released from the interna and last between 12 and 16 h; N3 appear between 12 and 16 h after release; N4 appear between 28 and 32 h; and N5 appear between 44 and 48 h. The cypris stage appears between 108 and 112 h. The redescribed morphologies allowed us to identify new characters. Antennular setation discriminates naupliar instars 3, 4 and 5. Telson and furca morphologies discriminate all naupliar instars. Furthermore, we demonstrate that the speed of larval development is similar within a single brood and between broods from different specimens, suggesting synchronization of larval development. From precise monitoring of broods every 4 h, we demonstrate that the life cycle of S. carcini includes five instars of naupliar larvae instead of four. The morphological characters of the larvae discriminate these naupliar instars and allow the identification of S. carcini from other Rhizocephala species. S. carcini larvae develop synchronously. Consequently, they might be an informative model to study larval development in crustaceans.

Characterization of Arginine Kinase in the Barnacle Amphibalanus Amphitrite and Its Role in the Larval Settlement.

Energy metabolism is a key process in larval settlement of barnacles, but the underlying molecular mechanisms remain ambiguous. Arginine kinase (AK) mainly participates in energy metabolism in invertebrates. So far, its roles in barnacles have not been studied. In the present study, we raised an antibody against AK from Amphibalanus amphitrite Darwin to characterize the roles of AK in the larval settlement process. Among the developmental stages, AK was highly expressed during the cypris stage. Along with the aging process in cyprids, the level of AK decreased. The immunostaining results showed that AK was localized to muscular tissues in cyprids, including antennules, antennular muscles, and thoracic limbs. The larval settlement rate decreased and larval movement was inhibited in response to treatments with high concentrations of AK inhibitors (rutin and quercetin). These results demonstrated that AK was involved in the larval settlement of A. amphitrite through mediating energy supply in muscle tissues. Moreover, further analysis indicated that both the p38 MAPK and NO/cGMP pathways positively mediated the expression of AK in cyprids.

Effect of chlorination on barnacle larval stages: Implications for biofouling control and environmental impact

Continuous low-dose chlorination (CLDC, 0.2 ± 0.1 mg l−1) is practised to control mussel settlement in industrial cooling systems. However, its effect on larval barnacles is poorly understood. Experiments were carried out with larvae of the barnacle Amphibalanus reticulatus to assess: 1) chlorine toxicity to discrete naupliar stages, 2) effect of transient chlorine exposure of nauplii on subsequent larval development and 3) effect of chlorine on cypris settlement and metamorphosis. Individual naupliar stages showed different levels of sensitivity to chlorine exposures with the stage II nauplii being the most susceptible to in-use levels (0.2 ± 0.1 mg l−1) of chlorination. Short term exposures (3–20 min) to 0.2 mg l−1chlorine did not prevent larval settlement. A significant percentage of treated cyprid larvae (non-feeding stage) survived, settled and metamorphosed to juveniles. A fraction of the chlorine-treated cypris larvae precociously metamorphosed into juveniles, bypassing cementation to substratum. About half of the pre-competent larvae (naupliar stages) exposed to in-use levels of chlorine failed to reach cypris stage, indicating that CLDC can impact barnacle larval development in the sea. In barnacle dominated systems, use of alternative control strategies/biocides may be required.

Larval development and settlement of the barnacle Amphibalanus amphitrite from the Red Sea: Influence of the nauplii hatching season

Abstract Barnacle Amphibalanus amphitrite adults were collected from the Jeddah coast of the Red Sea during different seasons. The nauplii released by adults in autumn, winter, spring and summer were reared under laboratory conditions to know the larval development duration and settlement in relation to the hatching season. The nauplii reared during winter (11 days) and autumn (13 days) took longer to reach the cypris stage compared to nauplii reared in summer (6 days) and spring (7 days). The most successful settlement of larvae was observed in spring and summer and the least successful — in winter. The observations of gonads showed that summer and spring are the active breeding season for A. amphitrite in the Red Sea. The results of this study indicated that the nauplii hatching season plays a significant role in the larval development and settlement of barnacles in the Red Sea.

A description of the larval development of Megabalanus azoricus (Pilsbry, 1916) reared in the laboratory

This study provides the first description of the larval development of the commercially exploited barnacle Megabalanus azoricus. It describes the changes in larval size and shape as well as the general morphology and duration of each larval stage. Embryos were obtained from gravid specimens collected at São Miguel Island and reared through six naupliar stages to the cypris stage in laboratory conditions. The planktotrophic nauplii reached the cypris stage after 14 days of hatching in individual cultures at 20 °C under natural illumination and fed with phytoplankton (Chaetoceros gracilis, Isochrysis sp., and Tetraselmis sp.). The nauplius of M. azoricus has a normal size compared with nauplii of other congeneric species, ranging between the 261 μm (nauplius I) and 912 μm (nauplius VI). This work provides the first description of larvae of the genus Megabalanus for the Portuguese oceanic islands and provides comparisons with congeneric species in other parts of the world.

Transcriptomic analysis of neuropeptides and peptide hormones in the barnacle Balanus amphitrite: evidence of roles in larval settlement.

The barnacle Balanus amphitrite is a globally distributed marine crustacean and has been used as a model species for intertidal ecology and biofouling studies. Its life cycle consists of seven planktonic larval stages followed by a sessile juvenile/adult stage. The transitional processes between larval stages and juveniles are crucial for barnacle development and recruitment. Although some studies have been conducted on the neuroanatomy and neuroactive substances of the barnacle, a comprehensive understanding of neuropeptides and peptide hormones remains lacking. To better characterize barnacle neuropeptidome and its potential roles in larval settlement, an in silico identification of putative transcripts encoding neuropeptides/peptide hormones was performed, based on transcriptome of the barnacle B. amphitrite that has been recently sequenced. Potential cleavage sites andstructure of mature peptides were predicted through homology search of known arthropod peptides. In total, 16 neuropeptide families/subfamilies were predicted from the barnacle transcriptome, and 14 of them were confirmed as genuine neuropeptides by Rapid Amplification of cDNA Ends. Analysis of peptide precursor structures and mature sequences showed that some neuropeptides of B. amphitrite are novel isoforms and shared similar characteristics with their homologs from insects. The expression profiling of predicted neuropeptide genes revealed that pigment dispersing hormone, SIFamide, calcitonin, and B-type allatostatin had the highest expression level in cypris stage, while tachykinin-related peptide was down regulated in both cyprids and juveniles. Furthermore, an inhibitor of proprotein convertase related to peptide maturation effectively delayed larval metamorphosis. Combination of real-time PCR results and bioassay indicated that certain neuropeptides may play an important role in cypris settlement. Overall, new insight into neuropeptides/peptide hormones characterized in this study shall provide a platform for unraveling peptidergic control of barnacle larval behavior and settlement process.

First report of antennular attachment organs in a barnacle nauplius larva

AbstractLarvae released from Newmaniverruca albatrossiana were cultured in the laboratory until the cypris stage. The brood size of individuals was low, about 60 larvae per brood. The exact number of instars was not determined. Early instars had the morphology normally seen in lecithotrophic nauplii of thoracican cirripedes. They had uniramous antennules with a few apical setae and biramous antennae and mandibles equipped with natatory setae only. Neither antennae nor mandibles carried any enditic spines or setae and the mouth cone was diminutive. The last nauplius stage obtained in our cultures was typical except in the structure of antennules. The head shield was enlarged but not flexed down, the antennae and mandibles were virtually unchanged from earlier instars, and the ventral thoracic process was well developed but without any external appendages. In contrast, the antennules had the complex shape and segmentation otherwise seen only in cypris larvae, where they are used for bipedal walking on the substratum in search of a settlement site. The similarity included the specialized shape of the first two antennular segments and the specialization of the third as an attachment organ. Nauplii just prior to this last instar had simple, straight antennules but completely lacked setae and instead terminated bluntly in what appears to be an incipient attachment organ. The presence of cypris‐like antennules in late nauplii has not previously been recorded in cirripedes. We suggest that this will allow the larvae to attach on the substratum temporarily before they reach the cypris instar and this will increase the chance of settling successfully on their rare substratum (sea urchin spines). The specialization in late N. albatrossiana nauplii will therefore decrease mortality during the larval phase and thus counterbalance the very low breeding potential in this deep‐sea species.

Larval development and settlement of a whale barnacle

Larval development and settlement of whale barnacles have not previously been described, unlike intertidal barnacles. Indeed, the mechanisms of the association between barnacles and whales have not been studied. Here we describe the larval development and settlement of the whale barnacle, Coronula diadema, and possible involvement of a cue from the host in inducing larval settlement. Eight-cell stage embryos were collected from C. diadema on a stranded humpback whale, incubated in filtered seawater for 7 days, and nauplius larvae hatched out. When fed with Chaetoceros gracilis, the nauplii developed to stage VI, and finally metamorphosed to the cypris stage. The larval development looked similar to that of intertidal barnacles with planktotrophic larval stages. The cyprids did not settle in normal seawater, but did settle in polystyrene Petri dishes when incubated in seawater with a small piece of skin tissue from the host whale. This strongly suggests the involvement of a chemical cue from the host whale tissue to induce larval settlement.

Larval development of the barnacle Ibla cumingi (Cirripedia: Pendunculata: Iblidae) reared in the laboratory

Larvae of Ibla cumingi from Daya Bay, China were cultured in the laboratory. Larval development includes six naupliar stages and a non-feeding cypris stage following the ground patterns of cirripeds. Larvae reached the cypris stage in nine days at ∼25°C after hatching. Morphological features including the cephalic shield, frontal horns, labrum, abdominal process, antennules, antennae and mandibles in all nauplii were described and illustrated using light microscopy. A full morphological description of cyprid larvae was provided using scanning electron microscopy. Attempts were made to compare the difference between the larval development model of I. cumingi and that of Ibla species.

Larval development of the parasitic barnacle Heterosaccus papillosus (Cirripedia: Rhizocephala: Sacculinidae) studied under laboratory conditions

Larval development of the parasitic barnacle Heterosaccus papillosus (Cirripedia: Rhizocephala: Sacculinidae) is described from larvae reared in the laboratory. Lecithotrophic nauplii of H. papillosus reached the cypris stage in 3·5 days, at 22–23°C. The development included five naupliar and the single cypris stage, thus following the typical pattern in the Rhizocephala Kentrogonida. Like the nauplii of all sacculinids, those of H. papillosus had a single pigmented nauplius eye, unsegmented frontolateral horns each with two long spines and a tubercle between the furcal rami, but they lack a flotation collar and have no seta on the antennal basipod. The male larvae are larger than female ones, the mean lengths in the two sexes never overlap. The most interesting feature of this species are the morphological differences between the male and female larvae. In the male nauplii III to V the anterior body margin is noticeably convex, that is not characteristic for female larvae. The male cyprid also has a more acuited anterior region, whereas that of the female is rounded.

Distribution, Adult Morphology, and Larval Development of Sacculina Sinensis (Cirripedia: Rhizocephala: Kentrogonida) in Hong Kong Coastal Waters

Abstract Sacculina sinensis (Cirripedia: Rhizocephala) is a parasitic barnacle first reported in Hong Kong by Boschma (1933) to infect the intertidal crab, Leptodius exaratus. Since then, no studies have been conducted to investigate its distribution and morphology, which are important in providing background information for further taxonomic and ecological studies. In the present study, S. sinensis has shown to have a limited distribution in Hong Kong, being confined to only four out of the nine sheltered boulder shores sampled and with a mean infestation rate of 11.3 percent. The localized distribution of S. sinensis in Hong Kong is probably due to its short larval development times (four naupliar stages and one cypris stage, completed in about six days) and hence limited larval dispersal distance. The externa of S. sinensis is ovoid in shape with two distinct shoulders, and the outer cuticle surface is spinous. From histological studies, the male receptacles are located posteriorly in the visceral mass...

Toxic effects of copper on larval development of the barnacle Balanus amphitrite

Experiments were conducted to assess the effects of copper on larval development in the barnacle Balanus amphitrite. In the first experiment, we compared the sensitivity of three naupliar stages to copper stress. Molting inhibition occurred at copper concentrations ranging from 32 μg l −1 in nauplius II to 128 g l −1 in nauplius VI. EC 50 for molting ranged from 97 μg l −1 in nauplius II to 129 g l −1 in nauplius VI. Decreased survival occurred at 128 μg Cu l −1 in all of the naupliar stages tested, with LC 50 ranging from 145 in nauplius II to 213 μg l −1 in nauplius VI. In the second experiment, we examined effects of copper on the development from nauplius II to cyprid. The larvae reached cypris stage only in treatments of ⩽16 μg Cu l −1. Our study therefore showed that molting was a more sensitive endpoint than survival, nauplius II was the most sensitive naupliar stage, and that whole larval development assay was more sensitive than assays using a particular larval stage. The results were discussed with respect to the use of this species in toxicity tests.

Combined effects of temperature and salinity on larval development and attachment of the subtidal barnacle Balanus trigonus Darwin

The combined effects of temperature and salinity on larval development and attachment of Balanus trigonus Darwin (Cirripedia, Balanidae) was examined under controlled laboratory conditions. Whilst larval survivorship was not affected (>70%), the duration of larval development was significantly affected by temperature and salinity. The effect of temperature was comparatively stronger than that of salinity. The majority of nauplius II larvae metamorphosed into cypris stage after 4–5 and 10–11 days at 28 °C (22–34‰) and 18 °C (22–34‰), respectively. Temperature, salinity and the duration of assay had a significant effect on cypris attachment with significant interaction among these main effects. Maximum (>80% in 6 days) and minimum percent attachment (0% in 6 days) on polystyrene surfaces were observed at 24 °C (34‰) and 18 °C (22‰), respectively. At high temperature (28 °C) and low salinity (22–26‰), larvae rapidly (4 days) developed into cyprids, but less than 33% attached. These results suggest that low larval attachment rates may lead to the low recruitment of B. trigonus in Hong Kong waters during summer when the water temperature is high (about 28 °C) and salinity is low (<26‰).

Larval Development of the Barnacle Chinochthamalus Scutelliformis (Cirripedia: Chthamalidae) Reared in the Laboratory

Abstract Larvae of Chinochthamalus scutelliformis (Darwin, 1854) (Cirripedia: Chthamalidae) from Hong Kong were cultured in the laboratory. Larval development includes six naupliar stages and a nonfeeding cypris stage following the ground patterns of Cirripedes. Larvae reached the cypris stage in 20 days at ∼21°C compared to 13 days at ∼28°C. Morphological features including the cephalic shield, frontal horns, labrum, hind body, antennules, antennae, and mandibles in all nauplius and cypris stages were described and illustrated using a combination of light microscopy and scanning electron microscopy. Attempts were made to compare morphological differences between the nauplii and cyprid of C. scutelliformis with those of Chthamalus malayensis and Chamaesipho species. Observations of the nauplii of Chinochthamalus scutelliformis supported Foster's suggestion to move this species from the genus Chamaesipho to the genus Chinochthamalus.