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.     

Organization of Interna in Sacculina polygenea (Crustacea: Rhizocephala)

We performed histological studies on trophic and reproductive systems of colonial interna in Sacculina polygenea, a parasite of the coastal crab Hemigrapsus sanguineus. The trophic system that performs functions of absorption, accumulation, and transportation of nutrients from the hemolymph of the host comprises the trophic epithelium of distal canals and transporting trunks. The reproductive system of interna consist of nuclei (early stages of development of the primordia of externae) and the primordia of externae in later stages of development. It has been shown that during morphogenesis of the nucleus two primordia arise, a primordium of the externa itself and a primordium of its trophic system. In the primordium of the ovary, we found oogonia; early oocytes and vitellogenic oocytes were found in the ovaries of the late primordia of the externae. The damaging effects of the interna on the ovaries and testes of the crab host are discussed. Thus, we have found numerous elements of reproductive and trophic systems in the colonial interna of S. polygenea. The term individual is proposed to be used for the externa in rhizocephalan barnacles with its trophic system.     

On the fine structure of the regressing ecdysial glands of Carcinus maenas L. (Crustacea decapoda) parasitized by Sacculina carcini thompson

Summary The ecdysial glands (Y organs) of the crab Carcinus maenas regress in the presence of an external parasite, Sacculina carcini. This regression is more or less severe and may lead to complete autolysis. Three gradual stages in this involutionary process are described. In stage I, the gland cells are nearly normal. Nuclei and cytoplasmic organelles remain unchanged, but large vacuoles begin to appear. Stage II corresponds to more or less drastic nuclear pyknosis and cytoplasmic alterations. Myelin figures are large and numerous. Lysosomes and autophagic vacuoles with phosphatase activity are abundant. However, the general cellular architecture remains preserved. Stage III corresponds to irreversible cytolysis; nuclear envelopes and plasma membranes have disappeared. What remains is an accumulation of cellular debris becoming engulfed by circulating hemocytes. Not all of the gland cells of any given Y organ show the same degree of regression; degeneration is asynchronous.Structures seemingly corresponding to absorptive roots of the parasite are seen. Their lumen is coated with microvilli. The putative direct and indirect influences of the rhizocephalan parasite on its host are discussed. Our results on regressing Y organs of parasitized crabs are compared with those on regressing ecdysial glands of insects.Dedicated to the memory of Sir Francis Knowles, the first investigator to examine the ultrastructure of the Y organ of Carcinus maenas We wish to express our thanks to Professor Berta Scharrer for her critical advice     

Anatomy of virgin and mature externae of Loxothylacus texanus,parasitic on the dark blue crab Callinectes rathbunae (Crustacea: Cirripedia: Rhizocephala: Sacculinidae)

Rhizocephalan parasites are dioecious organisms, in that one or several dwarf males are implanted into the external part of the female parasite soon after it emerges from the interior of the host animal. The structure of the female externa and its resident males is crucial for understanding both the reproductive biology and the taxonomy of these specialized parasites. We use scanning electron microscopy and histological methods to study the anatomy of juvenile and the mature externae of the rhizocephalan barnacle Loxothylacus texanus parasitizing the blue crab Callinectes rathbunae. We put emphasis on the implantation of males and the histology of the female reproductive organs. In the virgin externae, male cyprids attach around a cuticular hood covering the mantle aperture, which is partially blocked by a plug of cuticle so only trichogon larvae, not cyprids, can access the mantle cavity. This resembles the situation known from Sacculina carcini. The mature externa is characterized by a visceral mass that contains the ovary, paired colleteric glands, a single male receptacle, but paired receptacle ducts. The proximal attachment of the visceral mass is located at some distance from the basal stalk, as is characteristic for the genus Loxothylacus. The internal anatomy of the mature externa of L. texanus is in most features similar to that seen in other species of the Sacculinidae, which comprises the majority of rhizocephalan species. However, the single receptacle creates a situation where the two implanted males cannot be kept separate as in most other rhizocephalans, but pass through spermatogenesis in a common chamber. This may have unknown effects on the reproductive biology such as male–male competition. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

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.     

Intravital and histological studies of the internal organization of the rhizocephalan barnacleSacculina polygenea

We performed a short-term in vitro cultivation of interna fragments of the rhizocephalan barnacleSacculina polygenea, a parasite of the shore crabHemigrapsus sanguineus, and a histological study of an interna isolated from the host organism. It is shown that the colonial interna ofS. polygenea includes an extensive trophic system (a “root” system), which performs the functions of absorption, accumulation, and transportation of nutrients, and a reproductive system represented by numerous externa primordia at different stages of development. Vitellogenic oocytes were found in the ovary of the late primordia of externae. Thus, we observed the colonial structure of interna inS. polygenea with numerous elements of the reproductive and trophic systems and characteristic signs of fractal organization.     

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.     

Reproductive Stage of the Life Cycle in the Rhizocephalan Barnacle Polyascus polygenea (Crustacea: Cirripedia)

The specific features of the reproductive stage of the life cycle have been studied in the rhizocephalan barnacle Polyascus polygenea, a parasite of the coastal crab Hemigrapsus sanguineus. It is shown that a single crab can bear 1 to 8 externae of P. polygenea. The fecundity of the parasite depends on the size of the externae and their number on the host and may reach as much as 50000 eggs for one externa. In Peter the Great Bay, Sea of Japan, this species repeatedly reproduces during the entire spring–autumn period; externae with developing embryos in the mantle cavity occur from May to September, and planktonic larvae occur from June to October. One externa produces during the season of reproduction no less than three generations of larvae. Thus, the reproductive strategy in P. polygenea comprises a three-stage cascade of reproduction: asexual reproduction via budding of the interna; the development of several generations of one or several externae; and several reproduction cycles of each externa. This allows the parasite to produce a very great number of larvae and ensures the parasitization of a significant proportion of the host crab population. The structure of the ovaries and oogenesis in rhizocephalans and free-living cirripede barnacles have many common features, which provides evidence for integration of these two groups within one monophyletic taxon.     

The Morphology of the Naupliar Stages of Sacculina carcini(Crustacea: Cirripedia: Rhizocephala)

A morphological study was carried out on the fournaupliar stages of Sacculina carciniusing mainly scanning electron microscopy. Frontal horns were present and throughout development the typical nauplius limbs remained simple and gnathobases were lacking. Such features are characteristic of other lecithotrophic barnacle nauplii. The presence of a vestigial ventral thoracic process was evident on the stage III nauplius and was even more prominent on the stage IV nauplius. These observations confirm that the rhizocephalan nauplius is close to the thoracican nauplius form and lend strong support for the retention of the Rhizocephala within the Cirripedia.     

Geographic and host‐mediated population genetic structure in a cestode parasite of the three‐spined stickleback

Comparative studies of genetic diversity and population structure can shed light on the ecological and evolutionary factors that influence host–parasite interactions. Here we examined whether geography, time and genetic variation in Alaskan three‐spined stickleback (Gasterosteus aculeatus Linneaus) hosts shape the population genetic structure of the diphyllobothridean cestode parasite Schistocephalus solidus (Müller, 1776). Host lineages and haplotypes were identified by sequencing the mitochondrial cytochrome b gene, and host population structure was assessed by Bayesian clustering analysis of allelic variation at 11 microsatellite loci. Parasite population structure was characterized according to allelic variation at eight microsatellite loci. Mantel tests and canonical redundancy analysis were conducted to evaluate the proportion of parasite genetic variation attributable to time and geography vs. host lineage, haplotype, and genotypic cluster. Host and parasite population structure were largely discordant across the study area, probably reflecting differences in gene flow, environmental influences external to the host, and genomic admixture among host lineages. We found that geography explained the greatest proportion of parasite genetic variation, but that variation also reflects time, host lineage, and host haplotype. Associations with host haplotypes suggest that one parasite genotypic cluster exhibits a narrower host range, predominantly infecting the most common host haplotypes, whereas the other parasite cluster infects all haplotypes equally, including rare haplotypes. Although experimental infection trials might prove otherwise, distributional differences in hosts preferentially infected by S. solidus could underlie the observed pattern of population structure.     

Microbial communities are indicators of parasite infection status

Growing evidence suggests that microbiomes have been shaping the evolutionary pathways of macroorganisms for millennia and that these tiny symbionts can influence, and possibly even control, species interactions like host–parasite relationships. Yet, while studies have investigated host–parasites and microbiomes separately, little has been done to understand all three groups synergistically. Here, we collected infected and uninfected Eurypanopeus depressus crab hosts from a coastal North Carolina oyster reef three times over 4 months. Infected crabs demonstrated an external stage of the rhizocephalan parasite, Loxothylacus panopaei. Community analyses revealed that microbial richness and diversity were significantly different among tissue types (uninfected crab, infected crab, parasite externae and parasite larvae) and over time (summer and fall). Specifically, the microbial communities from parasite externae and larvae had similar microbiomes that were consistent through time. Infected crabs demonstrated microbial communities spanning those of their host and parasite, while uninfected crabs showed more distinctive communities with greater variability over time. Microbial communities were also found to be indicators of early-stage infections. Resolving the microbial community composition of a host and its parasite is an important step in understanding the microbiome's role in the host–parasite relationship and determining how this tripartite relationship impacts coevolutionary processes.     

Cloning and expression of the engrailed.a gene of the barnacle Sacculina carcini

 Cirripedia (barnacles) constitute a crustacean monophyletic taxon which is very well defined by several synapomorphies. In particular, all cirripedes are composed of six thoracic segments, but are devoid of any complete abdominal segment. This body plan is preserved in the adult in non-parasitic groups, while the parasitic rhizocephalan cirripedes completely lose arthropodian segmentation at the adult stage. These traits make them a particularly favourable model for studying the formation and maintenance of segmental identity. For the above reasons, it seemed worthwhile to look at the segmentation gene engrailed in a cirripede. A complete engrailed.a cDNA was isolated from larvae of the rhizocephalan cirripede Sacculina carcini. Its expression was monitored during larval development by use of the monoclonal antibody MAb4D9 directed against the Drosophila homologous proteins. The Sacculina engrailed.a gene is expressed during the second and third larval stages in stripes within a posterior area corresponding to the presumptive trunk segments. Surprisingly, these stripes appear in a posterior to anterior sequence. Six engrailed.a stripes characterize the thoracic segments of the cirripedean ground plan. Received: 18 June 1998 / Accepted: 24 October 1998     

Modelling the biological invasion of Carcinus maenas (the European green crab)

This paper proposes a system of integro-difference equations to model the spread of Carcinus maenas, commonly called the European green crab, that causes severe damage to coastal ecosystems. A model with juvenile and adult classes is first studied. Here, standard theory of monotone operators for integro-difference equations can be applied and yields explicit formulas for the asymptotic spreading speeds of the juvenile and adult crabs. A second model including an infected class is considered by introducing a castrating parasite Sacculina carcini as a biological control agent. The dynamics are complicated and simulations reveal the occurrence of periodic solutions and stacked fronts. In this case, only conjectures can be made for the asymptotic spreading speeds because of the lack of mathematical theory for non-monotone operators. This paper also emphasizes the need for mathematical studies of non-monotone operators in heterogeneous environments and the existence of stacked front solutions in biological invasion models.     

Seasonal changes in a population of Sacculina carcini Thompson (Crustacea: Rhizocephala) in Scotland

On the basis of studies of seasonal changes in the infection levels of Sacculina carcini in Carcinus maenas a life history for Sacculina at Millport, Scotland is proposed. This life history differs from that proposed by Foxon (1940) in that the externa is considered only to live for one year, the interna subsequently regenerating a new externa. This life cycle receives support from observations on regenerating externa.     

Barnacles vs bullies: modelling biocontrol of the invasive European green crab using a castrating barnacle parasite

Invasive species raise concern around the globe, and much empirical and theoretical research effort has been devoted to their management. Integrodifference equations are theoretical tools that have been used to understand the spatiotemporal process of a species invasion, with the potential to yield insight into the possible biological control measures. We develop a system of integrodifference equations to explore the potential release of a castrating barnacle parasite Sacculina carcini to control spread and abundance of an invasive species, Carcinus maenas, the European green crab. We find that the parasite does not completely eradicate the green crab population, but has the potential to reduce its density. Our model suggests that the crab population is likely to outrun the spread of the parasite, causing two waves of invasion travelling at different speeds. By performing a sensitivity analysis, we investigate the effects of the demographic parameters on the speed of invasion. To conclude, we discuss the predicted outcomes for the European green crab, and other non-target hosts, of using the castrating barnacle as a biocontrol agent.     

Hyperparasitism of the cryptoniscid isopod Liriopsis pygmaea on the lithodid Paralomis granulosa from the Beagle Channel, Argentina

A total of 29,570 false king crab Paralomis granulosa were sampled from the Beagle Channel (54 degrees 51'S, 68 degrees 12'W), Tierra del Fuego, Argentina, between July 1996 and August 1998. Crab size varied from 6.8 to 111.2 mm carapace length (CL). A few crabs parasitized by the rhizocephalan Briarosaccus callosus were found; prevalences of externae (the rhizocephalan reproductive body) and scars (the mark left on the host after the death of the parasite) were 0.28 and 0.16%, respectively. Of 85 externae examined, 55 were non-ovigerous and 30 ovigerous. The cryptoniscid isopod Liriopsis pygmaea infested 36.5% of the B. callosus examined. The most abundant stage was the cryptonicus larva, accounting for 208 of the 238 L. pygmaea recovered. Cryptonisci showed a highly aggregated distribution. A total of 92.7% of cryptonicsci were recovered inside empty externae, suggesting that the latter were attractive to cryptonisci. Early subadult females of L. pygmaea were rare; only 3 individuals occurred inside 1 ovigerous externa. Eight late subadult and 18 adult females were found on 3 and 7 non-ovigerous externae, respectively; in addition, 1 aberrant late subadult was found on 1 ovigerous externa. In the Beagle Channel, the population of P. granulosa harbours 3 different parasites: the bopyrid isopod Pseudione tuberculata, which reaches highest prevalence at 10 to 20 mm CL, the rhizocephalan B. callosus, with highest prevalence at 20 to 40 mm CL, and the cryptoniscid isopod L. pygmaea, which mainly infests rhizocephalan on crabs >40 mm CL.     

A novel approach to parasite population genetics: Experimental infection reveals geographic differentiation,recombination and host‐mediated population structure in Pasteuria ramosa,a bacterial parasite of Daphnia

The population structure of parasites is central to the ecology and evolution of host‐parasite systems. Here, we investigate the population genetics of Pasteuria ramosa, a bacterial parasite of Daphnia. We used natural P. ramosa spore banks from the sediments of two geographically well‐separated ponds to experimentally infect a panel of Daphnia magna host clones whose resistance phenotypes were previously known. In this way, we were able to assess the population structure of P. ramosa based on geography, host resistance phenotype and host genotype. Overall, genetic diversity of P. ramosa was high, and nearly all infected D. magna hosted more than one parasite haplotype. On the basis of the observation of recombinant haplotypes and relatively low levels of linkage disequilibrium, we conclude that P. ramosa engages in substantial recombination. Isolates were strongly differentiated by pond, indicating that gene flow is spatially restricted. Pasteuria ramosa isolates within one pond were segregated completely based on the resistance phenotype of the host—a result that, to our knowledge, has not been previously reported for a nonhuman parasite. To assess the comparability of experimental infections with natural P. ramosa isolates, we examined the population structure of naturally infected D. magna native to one of the two source ponds. We found that experimental and natural infections of the same host resistance phenotype from the same source pond were indistinguishable, indicating that experimental infections provide a means to representatively sample the diversity of P. ramosa while reducing the sampling bias often associated with studies of parasite epidemics. These results expand our knowledge of this model parasite, provide important context for the large existing body of research on this system and will guide the design of future studies of this host‐parasite system.     

Parasitism of the rhizocephalan <Emphasis Type="Italic">Briarosaccus callosus</Emphasis> on the crab <Emphasis Type="Italic">Paralomis verrilli</Emphasis> (Crustacea: Lithodidae) in the waters of southeast Sakhalin

The crab Paralomis verrilli in the waters of southeast Sakhalin is infected by the parasitic rhizocephalan barnacle Briarosaccus callosus. The prevalence of parasitic infection was on the average 4.36% (6.29% for females, 3.28% for males) and varied between samples from none up to 14.9%. The degree of prevalence was not related to the average carapace width and the sex ratio of crabs in samples. Sterilization of female P. verrilli was caused by the B. callosus infestation or its consequences. No more than two parasite externae per crab were found. Crabs with two externae made up 5.6% of all infected specimens. The infestation of crab hosts with two B. callosus externae negatively influenced the growth of the externae. A positive relationship was found between the width of the crab carapace and the length of the parasitic externae. The survival rate of P. verrilli with either one or two parasite B. callosus externae did not differ substantially. The proportion of crabs with externae and those with “scars” (12.2% in our case) can be taken as the index of survival of the parasitized crabs.     

Déroulement des gamétogenèses chez les Crabes Carcinus maenas (L.) et C. mediterraneus Czerniavsky parasités par la Sacculine

A histological study of the gonads and androgenic glands of Carcinus parasitized with Sacculina carcini showed no correlation between the degree of infestation of the gonads by the roots of the parasite and the progressive inhibition of the course of male and female gametogeneses. Also, no correlation could be observed between the degree of degeneration of the androgenic glands and that of the testes.

Modifications in parasitized crab testes essentially involve a degeneration of the mesoderm tissue of the germinative zone; primary gonia become pycnotic after an abnormal development; germ cells already engaged in spermatogenesis become blocked in prophase of meiosis; and ultimately, the testis becomes empty. Progressive degeneration of the androgenic glands, after an initial hypertrophy that starts as early as the internal Sacculina stage, seems related to the disorganization of the neurosecretory centers of the host.

Arrest of spermatogenesis at a more or less advanced stage is probably the result of an imbalance in neurohormonal controls acting directly on the germinative zone or indirectly via the androgenic glands. At the level of the ovaries, parasitic infestation leads to inhibition of the second phase of vitellogenesis possibly due to an abnormality in the organization of the vitellogenic follicles. Hormonal imbalances related to the presence of the parasite appear to be responsible for these phenomena.