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.     

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.     

Thompsonia dofleini,a colonial akentrogonid rhizocephalan with dimorphic,ova- or sperm-producing,externae (Crustacea,Cirripedia)

Summary Specimens of the blue swimming crab,Portunus pelagicus, are often infected with many thousands of externae ofThompsonia dofleini, all of which are connected through a common root system within the host crab. The species is unique in that the production of sperm cells takes place within the visceral mass of a small minority of the population of the externae. Spermatogonia are probably introduced by male cyprids into these externae when they are young, and they multiply and develop at the expense of the oocytes which rapidly disintegrate and ultimately disappear. It is assumed that the sperm cells are transferred to the ovary of the ordinary, egg-producing externae through the root system. Shortly after the eggs have been fertilized within the ovary they are transferred to the mantle cavity where they develop into cyprid larvae. The larvae become liberated when the externae drop off and the mantle wall disintegrates.Abbreviations used in the figures a annulus - an antennule - bm basal membrane around ovary - ce compound eye - cg cerebral ganglion - cr connecting root - cy cyprid larva(e) - c1 cuticle 1 - c2 cuticle 2 - do degenerating oocytes - e eggs in early division - ec embryonic cells - em embryos - en endocuticle of host - ep epidermis of host - ex exocuticle of host - hc hemocoelic cavity - m mantle - mc mantle cavity - mcy male cyprid - o ovary - oo oocyte - r rupture in wall of visceral mass - rs root system - sc scar - se spawned eggs - so spent ovary - sp spermatogonia - ss sperm and spermatids - st stalk - tc thickened cuticle of mantle cavity - th thorax - vm visceral mass - y yolk granules     

The anatomy and development of the rhizocephalan barnacle Clistosaccus paguri Lilljeborg and relation to its host pagurus bernhardus (L.)

The anatomy of all developmental stages of Clistosaccus paguri Lilljeborg including internal primordia only 200 μm across has been studied by conventional and ultrastructural techniques. The earliest primordia lack any roots and organs. A receptacle and ovary arise as separate structures. The colleteric gland develops by infolding of the mantle cavity epithelium. The primordium becomes external during a host intermoult. The externa is apparently female. It is proposed that male cyprids, through the integument, have implanted cells found in the mantle of the externa. These male cells travel through the mantle into the single receptacle, where they proceed with spermatogenesis. Externae lacking male cells will not grow. The larvae are released as cyprids, in which no preformed kentrogon is present. About half of the infected hosts carry more than one externa. Multiple externae are separate and may represent different invading female cyprids. C. paguri infects all sizes of its host, and castrates it. The effect of the parasite on the host is discussed. The life span of the parasite seems to be one year and the parasite population shows an annual cycle.     

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.     

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.     

Morphology and biology of Polysaccus japonicus (Crustacea, Rhizocephala, Akentrogonida, Polysaccidae, fam. n.), a parasite of the ghost-shrimp Callianassa japonica

The morphology and biology of Polysaccus japonicus Høeg & Lützen, 1993 is described. Nearly all infected hosts are adults, the females of which become castrated. Each bears 4–50 (female) externae on the underside of the abdomen. The externae are interconnected via a root system within the host's perineural blood sinus. The externae pass through three moults. Cypris larvae inject spermatogonia into the immature externa's mantle cavity, which is where spermiogenesis occurs later. Multiplication of the male cells is probably supported by secretion from paired mantle cavity glands, presumably homologous to the receptacles of kentrogonid rhizocephalans. Late during the 2nd intermoult the visceral mass ruptures to release the ova into the mantle cavity where they become fertilized. The third moult results in opening of the mantle aperture which initiates a period of vigorous peristalsis of the muscular mantle. Following emission of the cypris larvae the externae perish leaving distinct scars. The root system always contains buds of various sizes and stage of development. When the scarred host moults, the most advanced buds emerge to give rise to a new generation of externae. This is thought to be repeated for the rest of the host's life.
The antennule of the cyprid has a relatively very long and slender 3rd segment, a unique construction among Akentrogonida. Because of this and the presence of special mantle cavity glands, a new family, Polysaccidae, is erected, which in some characters is intermediate between the Kentrogonida and the Akentrogonida.     

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.     

Infestation Level of the Crab Hemigrapsus sanguineus by a Parasitic Crustacean Polyascus polygenea (Crustacea: Cirripedia) in Vostok Bay, Sea of Japan

We studied the infestation level of the coastal crab Hemigrapsus sanguineus by a cirripede barnacle Polyascus polygenea in Peter the Great Bay (Vostok Bay, Sea of Japan) in 1998–1999. It is shown that the externae of the parasite usually appear on the surface of the host’s body at the time when the crab attains sexual maturity (at a carapace width of 14–15 mm). More often we encountered infested crabs of a medium size, with a carapace width of 25–29 mm. The level of crab invasion did not depend on the sex of the host. In the summer period the proportion of infested crabs in different habitats ranged from 7 to 84%. The greatest number of crabs bearing the externae of the parasite was registered in an area with a boulder-pebble surface and small surf waves.Original Russian Text Copyright © 2005 by Biologiya Morya, Korn, Akhmadieva, Rybakov, Shukalyuk.     

Boschmaella japonica,New Species,a Parasite on the Barnacles Chthamalus challengeri and Balanus amphitrite amphitrite from Japan (Crustacea: Cirripedia: Rhizocephala: Chthamalophilidae)

Abstract A new rhizocephalan parasite which infests the barnacles Chthamalus challengeri and Balanus amphitrite amphitrite is described. It is known only from the type locality, Aburatsubo, Kanagawa Prefecture, Japan. Its position in the family Chthamalophilidae is assured by the cyprids lacking a thorax, by the position of the mesenteric canal opposite to the stalk, and by its host being a balanomorph barnacle. Among the three previously described species of the Chthamalophilidae, the new species most closely resembles Boschmaella balani, but it differs in details of the internal root system and in the externa being found both on the host body and on the inner mantle. About two-thirds of the infested hosts carry more than one externa.     

Control region sequences indicate that multiple externae represent multiple infections by Sacculina carcini (Cirripedia: Rhizocephala)

The rhizocephalan barnacle, Sacculina carcini, is a common parasite of the European shore crab, Carcinus maenas, in which it causes significant detrimental physical and behavioral modifications. In the vast majority of cases, the external portion of the parasite is present in the form of a single sac‐like externa; in rare cases, double or even triple externae may occur on the same individual host. Here, we use a highly variable DNA marker, the mitochondrial control region (CR), to investigate whether multiple externae in S. carcini represent infection by multiple parasites or asexual cloning developed by a single parasite individual. Sequences for multiple externae from C. maenas hosts from the Danish inlet, Limfjorden, and from the mud flates at Roscoff, France, were compared. In almost all cases, double or triple externae from an individual host yielded different haplotypes. In the few cases where identical haplotypes were identified from externae on a multiple‐infected host, this always represented the most commonly found haplotype in the population. This indicates that in Sacculina carcini, the presence of multiple externae on a single host reflects infection by different individual parasites. A haplotype network of CR sequences also suggests a degree of geographical partitioning, with no shared haplotypes between the Limfjorden and Roscoff. Our data represent the first complete CR sequences for a rhizocephalan, and a unique gene order was also revealed. Although the utility of CR sequences for population‐level work must be investigated further, the CR has proved a simple to use and highly variable marker for studies of S. carcini and can easily be applied to a variety of studies in this important parasite.     

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.     

The cypris larvae of the parasitic barnacle Heterosaccus lunatus (Crustacea, Cirripedia, Rhizocephala): some laboratory observations

Heterosaccus lunatus parasitizes the portunid crab, Charybdis callianassa in Moreton Bay, Australia. With the host crabs maintained at 22.5 degrees C this sacculinid rhizocephalan released larval broods every 6-7 days. During July-August 1996 and particularly August 1999 such broods showed the change-over from male only larvae in the early broods to females only in the later broods. As the host crabs were maintained under similar aquarium conditions in both years it is concluded that the light/dark cycle is the principal cue triggering this larval sex reversal. Oogenesis in the parasite externa is somehow controlled to produce two different sized ova - male larvae develop from large ova and females from small ova. A working hypothesis outlining how sex is probably determined for the larvae of sacculinids is erected. H. lunatus is considered the ideal sacculinid for the further experimental work necessary to verify the proposed sex-determining mechanism and its control processes. Measurements of the maximum swimming speeds of H. lunatus male and female cyprids showed the larger males to be the faster in absolute terms (27.95 compared with 17.60 mm s(-1), respectively), however, the calculated relative speeds were almost identical at approximately 90 body lengths s(-1). Settlement experiments confirmed that female H. lunatus cyprids settle only on the gills of C. callianassa; these cyprids needed to be at least 2 days old before they were able to settle.     

Cypris Settlement,Kentrogon Formation and Host Invasion in the Parasitic Barnacle Lernaeodiscus porcellanae (Müller) (Crustacea: Cirripedia: Rhizocephala)

The anatomy of the female cypris is very similar to that of cyprids of the Thoracica. There is no development of a preformed kentrogon within the unsettled cypris. Kentrogon formation is completed within 10–20 min after settlement. Elimination of the spent cypris is necessary for successful development. The kentrogon includes a variety of tissues, such as muscles and part of the cement glands. The injection stylet develops in connection with a moult, and it penetrates into the crab gill through the ventral body wall of the kentrogon c. 50 h after settlement. Only a single cell, also recognizable in the unsettled cypris, is injected into the host, while the rest of the kentrogon degenerates after invasion. The speed of kentrogen development and much of the kentrogon anatomy can be interpreted as a defense against host crab grooming. The single invasion cell may either be considered as a stem cell or an egg.     

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.     

Reproductive strategy of two deep-sea scalpellid barnacles (Crustacea: Cirripedia: Thoracica) associated with decapods and pycnogonids and the first description of a penis in scalpellid dwarf males

We investigated the sexual system in two pedunculate barnacles of the family Scalpellidae. Both inhabit deep water and are attached to mobile arthropod hosts. Verum brachiumcancri was attached to the majid crab Rochinia hertwigi, and Weltnerium nymphocola to the pycnogonidan sea spider Boreonymphon rubrum. Both barnacles have separate sexes and females almost always carry two dwarf males that are almost fully embedded in a pair of symmetrically situated receptacles inside the rim of the mantle cavity. The dwarf males of V. brachiumcancri have a complex penile structure extending into the female mantle cavity. This is the first time a copulatory structure has been described in detail for a dwarf male of a scalpellid barnacle. Both species lack free nauplii and their larvae are released as cyprids; the brood size is small. This is probably an adaptation for settling close to the parent population. We compare reproductive strategies among scalpellids and suggest that the present males are highly specialized and that the females are allocating resources to few offspring.     

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.     

Effects of temperature and salinity on the larvae of two species of rhizocephalan (Crustacea: Cirripedia)

Summary

Responses of larvae of two rhizocephalan species to changes in seawater temperature and salinity were studied under laboratory conditions. Peltogasterella gracilis parasitizes the hermit crab Pagurus pectinatus, which occurs at stable salinity and gradually changing temperature in summer. Sacculina polygenea is a parasite of the crab Hemigrapsus sanguineus, which lives in the intertidal zone in summer where salinity and temperature can fluctuate during the day. The development of both species is comprised of five naupliar stages and the cyprid stage, and it was considered successful if more than 50% of the nauplii attained the cyprid stage. P. gracilis nauplii successfully developed at 12–20°C and 30–34‰, but at 22°C successful development occurred in a narrower salinity range (32–34‰). All nauplii died both at 25°C and in 26‰. S. polygenea nauplii successfully reached the cyprid stage at higher temperatures (18–25°C) and a wider salinity range (18–34‰) than P. gracilis nauplii, but at 12°C and 16‰ larval development of S. polygenea was suppressed. Under favorable conditions, naupliar development lasted 3.5 days in P. gracilis and 2–3 days in S. polygenea. The cyprids of both rhizocephalan species demonstrated a greater resistance to temperature and salinity changes than nauplii. However, P. gracilis cyprids were active in a narrower salinity range (16–34‰), as compared to S. polygenea cyprids (8–34‰). Under favorable conditions the cyprids of both species survived for 6 to 10 days.     

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.