The cypris larvae of the rhizocephalan barnacle Heterosaccus lunatus with particular reference to antennular morphology

SEM investigation of laboratory-reared cyprids of the parasitic barnacle Heterosaccus lunatus has revealed morphological differences between the male and female larvae, particularly the attachment organ of their antennules. The detailed features of the attachment organ (segment III) and segment IV for each sex are described and compared with those of other rhizocephalan cyprids, particularly Sacculina carcini , a reportedly closely related species. The absence of the large posterior sac from segment III in the H. lunatus male highlights the need for careful larval investigations on other sacculinid species. Settlement of female H. lunatus cyprids occurred on the gill lamellae of the host crab with the subsequent kentrogon stage taking the sacculinid form. The inclusion of H. lunatus in the Sacculinidae was therefore confirmed, based on the morphology of the cypris antennules and kentrogon.     

Sex determination in the larvae of the parasitic barnacle Heterosaccus lunatus: an experimental approach

Heterosaccus lunatus parasitizes the portunid crab, Charybdis callianassa, in Moreton bay, Queensland, Australia. With the host crabs maintained at 25 °C this sacculinid rhizocephalan released larval broods every 5-6 days. This study examines the effect of photoperiod on the sex proportions of successive larval broods of individual parasites. Parasitized host crabs were maintained individually in circulating 25 °C seawater within two light-tight boxes, each with a controlled light-dark cycle operating. Box A had an LD 16:8 cycle (summer) and Box B an LD 8:16 cycle (winter). Both boxes had five host crabs and when parasites released their larval broods the larvae were flushed out and retained in a filter. Each captured brood then had its larvae sized to determine the proportions of the two sexes present. H. lunatus is ideal for such study because the sex(es) of all the larval stages (four naupliar stages and the cyprid) can be unequivocally identified by size (length). The experiment was run over a 6-week period during March and April 2004 when a total of 86 larval broods were released and their sex proportion(s) determined. It is concluded that larval sex determination is effected by photoperiod, with females becoming dominant under the winter condition (LD 8:16) and males dominant under the summer condition (LD 16:8). All initial broods were male dominated, so for Box A broods the male preponderance was maintained, whilst for Box B broods there was the progressive changeover to female preponderance. Such environmental sex determination is unusual because it is oogenesis which is being controlled, whereby two ovum sizes can be produced, either singly or together, the larger being the male egg. Further work is now necessary to verify the likely hormonal control processes involved.     

Size and settling behaviour in male and female cypris larvae of the parasitic barnacle Sacculina carcini Thompson (Crustacea: Cirripedia: Rhizocephala)

Twenty-one broods from different externae of Sacculina carcini Thompson were cultured to the cypris stage. The size of the cyprids was measured and the larvae subjected to settling upon unparasitized crabs (Garcinus maenas (L.)) and small juvenile externae. The cyprids occur in two sizes that may appear singly or together in the same brood. Small cyprids are of the female sex that settle upon crabs and are infective, while large cyprids are of the male sex and only settle upon juvenile externae. These results are in agreement with other well-studied rhizocephalans.     

Sex determination in the androdioecious barnacle Scalpellum scalpellum (Crustacea: Cirripedia)

How androdioecy (coexistence of hermaphrodites and males) is maintained is still poorly understood. Therefore, sex determination was studied in the androdioecious barnacle Scalpellum scalpellum L. First, 247 cypris larvae from seven broods were investigated for sexual dimorphism in larval morphology and found to be all identical. Second, experiments with cyprids showed that males and hermaphrodites differ distinctly in morphology as soon as 4–5 days after settlement. Third, 14 252 cyprids were allowed to settle on the bottom of their culture cages, and all surviving larvae developed into hermaphrodites and none into dwarf males. Fourth, larvae settled in hermaphrodite receptacles (i.e. future males) were removed at increasing intervals after settlement to study if the male and hermaphrodite sexual expressions are fixed or plastic. All larvae became dwarf males if allowed to stay there for more than 8 h after settlement. But if removed within 3 h after settlement, half of them developed into hermaphrodites. We conclude that an environmental sex determination mechanism operates in S. scalpellum. Together with a 1:1 hermaphrodite/male ratio observed in previously reported experiments offering a free choice of settlement, we suggest that all larvae are potential hermaphrodites, but only 50% can settle in hermaphrodite receptacles and yield males.     

Metamorphosis in the Cirripedia Rhizocephala and the homology of the kentrogon and trichogon

The Rhizocephala are considered to be monophyletic due to several synapomorphies in the ontogeny of the cndoparasitic phase. The various types of metamorphosis described in the Rhizocephala are discussed and compared to metamorphosis in the Cirripedia Thoracica and Acrothoracica. In males and females of the suborder Kentrogonida. the cyprid settles and metamorphoses into a new instar, in males the trichogen and in females the infective kentrogon. The kentrogon goes through yet another. incomplete moult associated with the development of the stylet. Within the three kentrogonidan families. the Iernaeodiscid-peltogastrid type of kentrogon differs from the sacculinid type in the mode of attachment to the host. in the complexity of internal anatomy. in the position and penetration of the stylet, and in whether or not the cyprid carapace must be shed prior to penetration of the stylet. In the Akentrogonida metamorphosis never results in a new instar. Where observed (Clistosaccidae and Thompsoniidae). both male and female cyprids settle and penetrate into their substrate (female parasite or new host) with one of the antennules. Using the antennule as a syringe. male cyprids inject spermatogonia while female cyprids injects embryonic cells developing into an endoparasite. By comparison with metamorphosis in the Cirripedia Thoracica and Acrothoracica it is concluded that the presence of a metamorphic moult leading to a post-cyprid instar is plesiomorphic and that the trichogon and kentrogon are homologous with the first metamorphosed juvenile in these outgroups. The abbreviated ontogeny in the Akentrogonida without metamorphic moult and post-cyprid larval instars is considered apomorphic. This contradicts the long-held supposition that the Akentrogonida are the most‘primitive’Rhizocephala and dovetails with new information that this suborder contains many advanced traits. Within the Kentrogonida. the lernacodiseid-peltogastrid type of kentrogon is considered more plesiomorphic than the sacculinid type, which resembles the clistosaccidthompsoniid type in having the antennules involved in the penetration process. The homologization of the kentrogon with a juvenile barnacle indicates that presence of a kentrogon is plesiomorphic within the Rhizocephala and that the Kentrogonida is paraphyletic.     

Bone-eating Osedax females and their 'harems' of dwarf males are recruited from a common larval pool

Extreme male dwarfism occurs in Osedax (Annelida: Siboglinidae), marine worms with sessile females that bore into submerged bones. Osedax are hypothesized to use environmental sex determination, in which undifferentiated larvae that settle on bones develop as females, and subsequent larvae that settle on females transform into dwarf males. This study addresses several hypotheses regarding possible recruitment sources for the males: (i) common larval pool--males and females are sampled from a common pool of larvae; (ii) neighbourhood--males are supplied by a limited number of neighbouring females; and (iii) arrhenotoky--males are primarily the sons of host females. Osedax rubiplumus were sampled from submerged whalebones located at 1820-m and 2893-m depths in Monterey Bay, California. Immature females typically did not host males, but mature females maintained male 'harems' that grew exponentially in the number of males as female size increased. Allozyme analysis of the females revealed binomial proportions of nuclear genotypes, an indication of random sexual mating. Analysis of mitochondrial DNA sequences from the male harems and their host females allowed us to reject the arrhenotoky and neighbourhood hypotheses for male recruitment. No significant partitioning of mitochondrial diversity existed between the male and female sexes, or between subsamples of worms collected at different depths or during different years (2002-2007). Mitochondrial sequence diversity was very high in these worms, suggesting that as many as 10(6) females contributed to a common larval pool from which the two sexes were randomly drawn.     

The relation between cypris ultrastructure and metamorphosis in male and female Sacculina carcini (Crustacea,Cirripedia)

Summary To elucidate current controversies on sex in rhizocephalan barnacles, broods of Sacculina carcini, infesting the shore crab Carcinus maenas, were raised to cyprids in the laboratory and followed through settlement and metamorphosis. Free-swimming cyprids were studied by transmission electron microscopy and occur in male and female morphological types, which differ in the structure of carapace cuticle, antennular cuticle, antennular glands, and the cells suspected of being the stem cells during metamorphosis. These dissimilarities are in addition to the already known differences in cypris size, in number of antennular sense organs, and in substrata settled on by morphological males and females. Metamorphosing males (trichogons) and females (kentrogons) are illustrated in interference phase-contrast micrographs. The morphological differences between male and female cyprids are directly related to their dissimilar metamorphosis. Hence, cyprids of male morphology are anatomically incapable of metamorphosing into kentrogons, while cyprids of female morphology cannot metamorphose into trichogons. The determination of sex in rhizocephalan barnacles is discussed.The results refute the hypothesis that sex in Sacculina carcini is determined environmentally, e.g., by the substratum encountered by the cyprids at settlement. It is concluded that sex is determined already in the free-swimming larvae and, most probably, already in the ovary. This agrees with the mode of sex determination in other species of the Rhizocephala Kentrogonida.     

The effect of host size on the sex ratio of Syngaster lepidus, a parasitoid of Eucalyptus longhorned borers (Phoracantha spp.)

The solitary larval ectoparasitoid, Syngaster lepidus Brullé, parasitizes the cryptic larvae of two wood-boring beetles, Phoracantha recurva Newman and Phoracantha semipunctata F. The objective of this study was to determine how the female parasitoids allocated the sex of progeny when presented with larval hosts of uniform size classes. Host size was directly correlated with age of the Phoracantha larval hosts. Groups of Phoracantha larvae of a single age class (2-, 3-, 4-, or 5-week-old) were exposed to parasitoids, and sex ratios of the resulting parasitoid progeny from each host age class were determined. A significant relationship was observed among the sizes of P. recurva and P. semipunctata hosts and the sex ratio of emerging parasitoids. Parasitized 2-week-old beetle larvae of both Phoracantha spp. produced only male S. lepidus progeny, whereas older larval hosts produced increasing proportions of female parasitoids (up to 80% females from 5-week-old hosts). Two-week-old Phoracantha larvae of both species produced fewer parasitoids than host larvae 3–5-week-old. The size of parasitoid progeny consistently increased with host larval age (size), and female parasitoids were larger than males across all host size classes. Male S. lepidus developed in approximately 25 days from 2-week-old hosts, and 19–21 days in 3–5-week-old hosts. Female S. lepidus developed in 22–25 days, with developmental time increasing with host size.     

Maternal adjustment of the sex ratio in broods of the broad-horned flour beetle, Gnathocerus cornutus

We report that females of the broad-horned flour beetle, Gnathocerus cornutus, can plastically adjust the sex ratio in their broods in response to environmental quality. Specifically, females reared in nutritionally poor environments produce broods that are 65% female, on average, with the degree of female-bias in some broods approaching 95%. In addition, females reared in nutritionally poor environments lay significantly more eggs than do females reared on standard medium, which produce broods with an even sex ratio. These effects of the mother's environment on size and sex ratio in broods are manifest even when oviposition occurs in the standard nutritional environment; indeed, the degree of female-bias increases with advancing female age despite the availability of nutritional resources to females at the time of egg laying. Our studies rule out sex-specific differences in viability early in larval development as the mechanism for the bias in sex-ratio of broods, since females reared in nutritionally poor environments have broods with hatchability and larval viability comparable to those of nonstressed females. Our studies also rule out an effect of the sire on the sex ratio in broods, since all male mates were reared on standard medium. We discuss our results in the context of theories for the evolution of plastic sex-ratios in the face of environmental deterioration and discuss how plasticity can resolve a long-standing question about the conditions underlying the evolution of biased sex ratios.     

Competing for last place: Mating behaviour in a pill-box crab, Halicarcinus cookii (Brachyura: Hymenosomatidae)

The mating strategy of Halicarcinus cookii was investigated to ascertain how males maximised their fitness through mate choice. An intertidal population at Kaikoura, New Zealand, was dominated by mature crabs of both sexes in summer and by immature crabs in the colder months. More than 95% of mature females were ovigerous with early stage and late stage broods found in almost every month, indicating that egg production and larval release is continuous. The operational sex ratio was less than 1 male/female in summer, but often more than 1.0 in the colder months. The gonosomatic index increased along with brood development so that as soon as zoeae were released, the next clutch of eggs was ready to be fertilised. Males searched for receptive females and began pre-copulatory mate guarding without any courtship display. They mated preferentially with late stage or non-ovigerous females: copulation duration was longest for stage 5 females as was post-copulatory guarding (mean 18.3 h). Late stage females were up to 14% of the female population. Mate attraction seems to be the result of an ovarian signal rather than from the developing brood. Manipulation of the sex ratio had effects upon copulation duration and post-copulatory guarding: presence of a rival male increased duration of guarding. Females showed precocious mating in the penultimate instar and were able to lay fertilised eggs after their pubertal moult in the absence of males. H. cookii females have many mates, but males attempt to ensure paternity by preferentially pursuing mature females close to egg laying and by guarding these females after copulation. These behaviours are all elements of a competitive strategy to ensure that a male loses (not wins) the race to copulate because females have a ventral seminal receptacle, giving sperm precedence to the last male to mate. Male mating behaviour is a consequence and evolutionary response to female morphology.     

How whale and dolphin barnacles attach to their hosts and the paradox of remarkably versatile attachment structures in cypris larvae

How larvae of whale and dolphin epibionts settle on their fast-swimming and migrating hosts is a puzzling question in zoology. We successfully reared the larvae of the whale and dolphin barnacle Xenobalanus globicipitis to the cyprid stage. We studied the larval developmental ecology and antennular morphology in an attempt to assess whether an epibiotic lifestyle on this extreme substratum entails any unique larval specializations. Morphological parameters were compared with five other barnacle species that also inhabit extreme substrata. We found no larval specializations to a lifestyle associated with marine mammals. The external morphology of the antennules in Xenobalanus cyprids is morphologically similar to species from strikingly different substrata. We found variation only in the structures that are in physical contact with the substratum, i.e., the third segments carrying the villi-covered attachment disc. The third segments of the Xenobalanus cyprid antennules are not spear-shaped as in the stony coral barnacles, which are here used to penetrate the live tissue of their hosts. The presence of a cyprid cement gland implies that Xenobalanus uses cement protein when attaching to its cetacean host. Naupliar instars developed outside of the mantle cavity, indicating dispersal is planktonic. Our results militate against the idea that the cyprids settle during ocean migrations of their hosts. We suggest cyprids settle during coastal aggregations of the cetacean hosts. We conclude that the ecological success of barnacles has ultimately depended on a larva that with little structural alteration possesses the ability to settle on an amazingly wide array of substrata, including cetaceans.

     

Host sex preferences and transmission success by the water mite Unionicola foili (Acari: Unionicolidae) parasitic on the midge Chironomus tentans (Diptera: Chironomidae)

This study examined whether ecoparasitic larval Unionicola foili exhibited a sex bias when infecting laboratory populations of the host insect Chironomus tentans and whether an association with male or female midges increased the likelihood of larval mites returning to the aquatic habitat. When laboratory populations of C. tentans were exposed to larval U. foili, there was a higher prevalence of mites among female hosts at emergence (17 of 30 males vs. 25 of 30 females infected by mites). However, there was no significant difference in the distribution or abundance of larvae among infected male (mean = 2.3 larvae per host) and female (mean = 2.6 larvae per host) midges. Larval mites parasitizing both male and female chironomids were more likely to return to water than could be expected by chance. Mite larvae infesting female C. tentans were more likely to return to water when female hosts deposited egg masses in water, suggesting that oviposition plays an important role in cueing larvae parasitizing female midges to detach. The mechanism responsible for increasing the likelihood that mites parasitizing male hosts return to water remains unclear. Future studies will address the possibility of parasite-mediated changes in host behavior.     

Costs and consequences of superparasitism in the polyembryonic parasitoid Copidosoma koehleri (Hymenoptera: Encyrtidae)

Abstract.  1. Polyembryonic wasps provide dramatic examples of intra-specific developmental conflict. In these parasitoids, each egg proliferates into a clonal lineage of genetically identical larvae. If more than one egg is laid in a host (superparasitism), individuals of different clones may compete for food resources.
2. In the polyembryonic encyrtid Copidosoma koehleri , one larva per clone can differentiate into a sterile soldier. It is shown that soldiers are always females, and that they attack intra-specific competitors.
3. Research hypotheses were that (a) clones that develop in superparasitised hosts suffer heavier mortality than clones that develop in singly parasitised hosts, and (b) female clones cause higher mortality to their competitors than male clones, hence larval survival is lower in superparasitised hosts that contain females than in male-only broods.
4. The potential frequency of superparasitism in C. koehleri was manipulated by varying parasitoid–host ratios and exposure durations.
5. As parasitoid densities and exposure durations increased, the frequency of superparasitism rose, brood sizes increased, but the number of hosts that completed development was reduced. The number of offspring per parasitoid female decreased with increasing parasitoid–host ratios. Offspring size and longevity were inversely correlated with brood size. As superparasitism rates increased, fewer all-male broods were produced. Male–female broods were female-biased, suggesting selective killing of males by female soldiers. All-female broods were significantly smaller than all-male broods at high parasitoid densities only, possibly reflecting aggression among soldiers of competing clones.
6. The results support the working hypotheses, and suggest that female larvae outcompete males in superparasitised hosts.     

The contributions of larval growth and pupal duration to protandry in the black swallowtail butterfly,Papilio polyxenes

Summary Shorter male larval and pupal durations were sufficient to produce the protandrous emergence pattern of the black swallowtail butterfly. In 9 broods observed under natural conditions, the first male was seen 7.1±6.5 days before the first female, although the peak of captures relative to the first capture was similar in both sexes. The field data supported only weakly the predictions of the Wiklund-Fagerstrom model for protandry. Some prereproductive period was necessary for males, and they were limited in how frequently they could mate. A third of the females mated more than once.Males were significantly smaller in size and weight than females in both diapausing and non-diapausing broods. Feeding experiments indicated that male larvae ate less and converted their food into biomass more efficiently than females. Female pupae contained higher proportions of fat and protein than male pupae. Differential body composition may be involved in sexual dimorphism differences in both larval and pual developmental rates. Thus, in addition to size, chemical differences in composition or metabolism may contribute to the observed natural protandry.     

Larval Competition, Adult Fitness, and Reproductive Strategies in the Acoustically Orienting Ormiine Homotrixa alleni (Diptera: Tachinidae)

Homotrixa alleni is a gregarious endoparasitoid fly that attacks adult male Sciarasaga quadrata (Orthoptera: Tettigoniidae) in southwestern Western Australia. Gravid female flies acoustically orient to their host's call and deposit live first-instar larvae upon or near their calling host. Up to 16 larvae may be found developing in the one host, and since only calling adult male S. quadrata are parasitized, host size and hence larval resources are essentially fixed at parasitism. This study examines parasitism by H. alleni in relation to intraspecific larval competition and adult fitness. The mean number of larvae emerging per host failed to increase significantly beyond a clutch size of four. Mean pupal weight and survival to the adult stage decreased linearly with increasing clutch size across the entire range of clutch sizes examined. Within a clutch, heavier pupae successfully completed pupal development significantly more often than lighter pupae. Pupal weight was directly related to adult size, with adult males being significantly larger than adult females at any given pupal weight. Female body size was positively correlated with fecundity. The size distribution of emerging females was normally distributed, while the distribution of searching gravid females collected at acoustic traps in the field was significantly skewed toward larger flies, suggesting yet another fitness benefit associated with large size. Using fecundity and survival to adulthood as our measure of fitness we calculated the optimal clutch size maximizing fitness per host to be seven, which exceeds the majority of observed clutch sizes in the field. Uncertainties associated with larvae successfully entering the host following larviposition are likely to reduce clutch sizes of H. alleni below this optimum in the field.     

Ladies last: diel rhythmicity of adult emergence in a parasitoid with complementary sex determination

Protandry, the earlier adult emergence of males, is explained as either an adaptive strategy maximizing male mating opportunities at the same time as minimizing female pre‐reproductive mortality, or as an incidental by‐product of sexual dimorphism fuelled by selection for other life‐history traits. Adult emergence sequences are monitored of broods of the gregarious larval endoparasitoid Cotesia glomerata L. (Hymenoptera: Braconidae) undergoing pupal development under different temperature regimes. As a haplodiploid species with single‐locus complementary sex determination, gender in C. glomerata is determined by the genotype at one sex locus. Haploids are always male, whereas diploids are female when heterozygous but male when homozygous at the sex locus. Sibling mating promotes homozygosity and thus the production of diploid males. Diploid males are produced at the expense of females, and impose a genetic burden on individuals and populations, despite their exceptional fertility in C. glomerata. Emergence of broods is typically completed within 2 days. Irrespective of temperature, males emerge earlier and within a shorter time interval than females, and a majority of the males in a cluster emerge before the first female. The implications of an incomplete temporal segregation of the sexes on the incidence of inbreeding in C. glomerata are discussed in the light of its sex determination mechanism and its patterns of mating, host exploitation and natal dispersal.     

Action of a rhizocephalan on the genital activity of host male crabs: characterization of a parasitic secretion inhibiting spermatogenesis

Protein fractions obtained by filtration of crude extracts of roots from the sacculinid parasite Loxothylacus panopei were tested by repeated injections on healthy male crabs Rhithropanopeus harrisii, the usual host of the rhizocephalan. Testes, and rogenic glands and different parts of the host CNS were observed; the behaviour of the animals was also noted. For the first time, proteinaceous substance(s) of about 25000–30000 daltons were characterized. They induce inhibition of spermatogenesis, and rogenic gland cytolysis and depletion of the sinus gland as in naturally infested crabs. The mode of action of sacculinid on spermatogenesis in the host is discussed.     

Host specificity of <Emphasis Type="Italic">Sacculina carcini</Emphasis>, a potential biological control agent of the introduced European green crab Carcinus maenas in California

The European green crab, Carcinus maenas, is an introduced marine predator established on the west coast of North America. We conducted laboratory experiments on the host specificity of a natural enemy of the green crab, the parasitic barnacle Sacculina carcini, to provide information on the safety of its use as a possible biological control agent. Four species of non-target, native California crabs (Hemigrapsus oregonensis, H. nudus, Pachygrapsus crassipes and Cancer magister) were exposed to infective larvae of S. carcini. Settlement by S. carcini on the four native species ranged from 33 to 53%, compared to 79% for green crabs. Overall, cyprid larvae tended to settle in higher numbers on individual green crabs than on either C. magister or H. oregonensis. However, for C. magister this difference was significant for soft-shelled, but not hard-shelled individuals. Up to 29% of the native crabs arrested early infections by melanizing the rootlets of the parasite. Most native and green crabs settled on by S. carcini became infected, especially when settled on by >3 cyprids. Infected green crabs died at more than twice the rate of uninfected green crabs. In contrast to green crabs, all infected native crabs died without producing an externa (reproductive sac). At high settlement intensities, infected native crabs frequently exhibited neurological symptoms (twitching, loss of movement) before death. These results indicate that use of S. carcini as a biological control agent could result in the death of native crabs. The magnitude of this effect would be proportional to the density of infected green crabs in the environment and the probability that cyprids would contact native crabs in the wild. Potential benefits of biological control should be assessed in relation to these potential non-target effects.     

Differences in the parasitic effects of a bopyrid isopod and rhizocephalan barnacle on the portunid crab,Charybdis bimaculata

Bopyrid isopods and rhizocephalan barnacles are obligate parasite crustaceans which harm their decapod hosts. However, to the best of our knowledge, studies have not compared which of these parasites has a greater parasitic effect on its hosts. Here, the parasitic effect of the bopyrid isopod, Allokepon hendersoni, and an unidentified sacculinid rhizocephalan species, infesting the same population of portunid crabs, Charybdis bimaculata, was investigated and compared for the first time. Samples were collected from the bycatch of a trawl fishery in Tosa Bay, Japan. A total of 2601 crabs were collected, of which 14 (0.55%) were parasitized by the bopyrid and 21 (0.82%) by the rhizocephalan. One of the two female crabs parasitized by the bopyrid was ovigerous (with much fewer eggs than unparasitized females). No ovigerous crab was found from the eight females parasitized by the rhizocephalan. Because only two female crabs were parasitized by the bopyrid, the following analyses were made using the male crabs. Both parasites reduced the wet weight (crab condition) and the cheliped size (secondary growth) of C. bimaculata, but the impact of the parasitism did not differ between the parasite species. The size of the abdominal flap of male hosts was reduced by the bopyrid infestation; however, rhizocephalan infestation caused enlargement of the abdominal flap, which is an indication of feminization. The present study provides information on how the effect of these two parasitic castrators on the same host crab varies. A moderate decrease in crab condition and cheliped development was common among the parasites, suggesting that the degree of impact might be favorable for the survival of the two parasites 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.