Vertical and horizontal distribution patterns of the giant sea anemone Heteractis crispa with symbiotic anemonefish on a fringing coral reef

The distribution patterns of the leathery sea anemone, Heteractis crispa, which contains an algal endosymbiont (zooxanthellae) and anemonefish, were investigated in relation to size distribution on a shallow fringing reef (3.2 ha, 0–4 m depth) in Okinawa, Japan. Individual growth and movements were also examined. Large individuals (>1,000 cm²) inhabited reef edges up to a depth of 4 m, while small anemone (<500 cm²) inhabited shallow reefs including inner reef flats. Individuals rarely moved, and their sizes were significantly correlated with their water depths. Growth of small anemones was negatively correlated with their distance from the reef edge, suggesting that reef edges provide more prey and lower levels of physiological stress. This study suggested that deep reef edges are suitable habitats for H. crispa. Large anemones were inhabited by large Amphiprion perideraion or large Amphiprion clarkii, both of which are effective defenders against anemone predators. Anemones that settle in deep reef edges may enjoy a higher survival rate and attain a large size because of their symbiotic relationship with anemonefish. However, early settlers do not harbor anemonefish. Their mortality rate would be higher in the deep edges than in shallow edges, the complicated topography of which provides refuge.     

Influence of <Emphasis Type="Italic">Lactobacillus fermentum</Emphasis> I5007 on the intestinal and systemic immune responses of healthy and <Emphasis Type="Italic">E. coli</Emphasis> challenged piglets

The effect of feeding Lactobacillus fermentum I5007 on the immune system of weaned pigs with or without E. coli challenge was determined. Twenty-four weaned barrows (6.07 ± 0.63 kg BW) were randomly assigned to one of four treatments (N = 6) in a factorial design experiment. The first two treatments consisted of healthy piglets with half of the pigs receiving no treatment while the other half was orally administered with L. fermentum I5007 (10⁸ CFU/ml) at a daily dose of 20 ml. Pigs in the second two treatments were challenged on the first day with 20 ml of E. coli K88ac (10⁸ CFU/ml). Half of these pigs were not treated while the remaining pigs were treated with 20 ml of L. fermentum I5007 (10⁸ CFU/ml). Peripheral blood lymphocytes subsets were determined using flow cytometry. The intestinal mucosal immunity of the pigs was monitored by real time polymerase chain reaction. The cytokine content of the pig’s serum was also analyzed. Oral administration of L. fermentum I5007 increased blood CD4⁺ lymphocyte subset percentage as well as tumor necrosis factor-α and interferon-γ expression in the ileum. Pigs challenged with E. coli had elevated jejunal tumor necrosis factor-α while interferon-γ expression was increased throughout the small intestine. There was no difference in the concentration of the cytokines interleukin-2, interleukin-6, tumor necrosis factor-α and interferon-γ in the serum. CD8⁺ and CD4⁺/CD8⁺ in peripheral blood were not affected by treatment. In conclusion, L. fermentum I5007 can enhance T cell differentiation and induce ileum cytokine expression suggesting that this probiotic strain could modulate immune function in piglets.     

Larvae and juveniles of the plunderfishes <Emphasis Type="Italic">Artedidraco shackletoni</Emphasis> and <Emphasis Type="Italic">A.</Emphasis><Emphasis Type="Italic">skottsbergi</Emphasis> (Notothenioidei,Artedidraconidae) from the Indian sector of the Southern Ocean

Early life stages of Artedidraco skottsbergi and A. shackletoni were collected off Adélie Land. The morphology and pigmentation pattern of nine larvae and juveniles of A. skottsbergi between 17.2 and 21.4 mm in standard length (SL), and of two juveniles of A. shackletoni measuring 25.1 mm SL were described. A. skottsbergi was characterized by a heavily pigmented body, except for the caudal peduncle, with distinctively dense pigmentation on the ventrolateral half of the body and caudal section (17.2–17.9 mm SL). Furthermore, they had no pigmentation on the pectoral fin base until they attained 21.4 mm SL. Juvenile A. shackletoni had a heavily pigmented body except for the ventral side of the abdomen and the anal fin base. The proximal part of the dorsal fin and most of the anal fin were covered with melanophores. Although knowledge of larval and juvenile Artedidraco species is limited, the distribution of melanophores on the fins, pectoral fin base and caudal peduncle at each developmental stage may be useful for species identification.     

Homologies of cephalic lateral line canals in <Emphasis Type="Italic">Pseudorhombus pentophthalmus</Emphasis> and <Emphasis Type="Italic">Engyprosopon grandisquama </Emphasis>(Pleuronectiformes): innervation and upper eye floor formation

Cephalic lateral line canals in two pleuronectiforms, Pseudorhombus pentophthalmus (Paralichthyidae) and Engyprosopon grandisquama (Bothidae), were studied and their homologies between the ocular and blind sides assessed on the basis of position and innervation patterns. A blind side canal, comprising small ossicles in a line lateral to the upper eye floor, was confirmed as the infraorbital line because the canal was not innervated by a ramus associated with the upper nasal (i.e., the superficial ophthalmic ramus innervating the supraorbital line). Consequently, the ramus innervating the canal was identified as the buccal ramus (associated with the infraorbital line). The blind side frontal forming the posterior half of the upper eye floor was identified as that part bearing the anteriormost otic canal in the ocular side, hypertrophy of the blind side component being evident. The supraorbital line of the blind side was represented by the upper nasal only in E. grandisquama.     

In-stream behaviour of threatened fishes and their food organisms based on remote video monitoring

Can remote underwater video be used to investigate the in-stream behaviour of small fishes and decapods? Diel activity of two threatened freshwater fishes (Macquaria australasica and Gadopsis bispinosus), a palaemonid prawn and an atyid shrimp, was established from remote underwater video in a pool of an upland stream in the current study. Decapods and large fishes (>5 cm TL) were nocturnal, whereas, small fishes (<5 cm TL) were diurnal. The suitability of using underwater video to quantify short-term (seconds) behavioural interaction among individuals was also demonstrated, with early-juvenile-phase Gadopsis bispinosus exhibiting interference competition on 35% of occasions when two or more individuals were observed. This study demonstrates that remote underwater video is useful for observing the in-stream behaviour of threatened freshwater fauna where other techniques are not viable, and presents sub-sampling of video as a means of reducing video processing time in assessing fish diel activity patterns.     

Coral settlement onto Halimeda opuntia: a fatal attraction to an ephemeral substrate?

Degraded reefs with a high abundance of macroalgae usually also have low densities of coral recruits. Few studies, however, have examined whether these algae affect coral larval settlement. This study demonstrates, experimentally, that larvae of the Caribbean coral Favia fragrum can settle on the green alga Halimeda opuntia even when another substrate more suitable for settlement is present. Larval settlement onto experimental substrates was quantified under three treatments: rubble only, rubble plus plastic algal mimic, and rubble plus live H. opuntia. Similar total larval settlement was observed in all treatments. No larvae settled on the algal mimic, but total settlement was similar on the rubble in the first two treatments, showing that the rubble alone offered sufficient substrate for high settlement success. About half the larvae in the live algal treatment settled on H. opuntia instead of on the rubble, showing that larvae did not reject this substrate as they did the algal mimic. This result raises the possibility that corals will settle on some macroalgae when their abundance is high. Most macroalgae, including H. opuntia, are ephemeral substrates unsuitable for post-settlement survival. Such unexpected settlement may therefore have significant consequences for coral recruitment success on algal-dominated reefs.     

Vaccination of sex reversed hybrid tilapia (Oreochromis niloticus × O. aureus) with an inactivated Vibrio vulnificus vaccine

Vibrio vulnificus causes disease in economically important aquaculture raised fish and is an opportunistic human pathogen. This study reports on the isolation of V. vulnificus from diseased hybrid tilapia (Oreochromis niloticus × O. aureus) cultured in a North American water reuse facility. Our objectives were to characterize the isolate using biochemical and molecular methods, develop a disease challenge model, and determine the ability of a formalin inactivated whole-cell vaccine to protect against V. vulnificus. The V. vulnificus isolate recovered was biotype 1, 16S rRNA type B, vcg type C, and vvhA type 2 and caused disease in tilapia held in static salt water (1.5 g/l sea salt). Fish vaccinated with the formalin inactivated whole-cell vaccine responded to vaccination with titers from vaccinated fish ranging from 32 to 64 and titers from non-vaccinated fish ranging from 4 to 8. In two trials, vaccinated tilapia exhibited relative percent survival (RPS) of 73 and 60% following homologous isolate challenge. In two additional trials, vaccinated tilapia exhibited RPS values of up to 88% following challenge with a heterologous isolate; the use of a mineral oil adjuvant enhanced protection. This vaccine may provide an effective means of preventing infections caused by biochemically and genetically diverse V. vulnificus.     

Temporal Fluctuations and Settlement Patterns of Native and Lessepsian Herbivorous Fishes on the Lebanese Coast (Eastern Mediterranean)

We provide here the first information on the abundance, settlement patterns and microhabitat use of settlers of two Lessepsian siganid species, Siganus rivulatus and S. luridus, in comparison with the two main native herbivores, Sparisoma cretense (Scaridae) and Sarpa salpa (Sparidae). S. rivulatus was the most abundant species (72%), followed by S. cretense (20%) and S. luridus (8%), whereas S. salpa was rare (<1%). A clear pattern of settlement could be established for S. rivulatus and S. cretense, with similar timing in 1999 and 2000. S. cretense settled in schools late June, while S. rivulatus settled mid-July. A few solitary settlers of S. luridus were observed in July and August, but none of S. salpa. All these herbivorous fish species settled in protected shallow areas offering hard substrates and algal communities. S. rivulatus was the most tolerant and adaptable species, able to settle on a large range of substrates and habitats, including rock pools, muddy harbours and sea grass beds. In the eastern Mediterranean Sea, S. rivulatus might have benefited from a release of competition pressure due to the low diversity and abundance of native herbivores. It has probably replaced S. salpa on the coast of Lebanon, being more competitive than the native sparid.     

Interannual variation in the diet of non-breeding male Antarctic fur seals, <Emphasis Type="Italic">Arctocephalus gazella</Emphasis>, at Isla 25 de Mayo/King George Island

The diet of non-breeding male Antarctic fur seals, Arctocephalus gazella, was investigated at Stranger Point, King George Island, through the analysis of scats during three consecutive summer seasons (1996, 1997, 1998). Overall, fish and krill were the most frequent prey occurring, respectively, in an average of 82.9% and 78.8% of samples (n = 131), followed by penguins (22.8%) and cephalopods (17.8%). Myctophids constituted almost 90% of the fish predated, with Electrona antarctica and Gymnoscopelus nicholsi being the most abundant and frequent species consumed. All fish taxa identified were krill feeding species suggesting that seals foraged mainly on a krill and a fish community associated with krill aggregations. However, a seasonal change was observed in the relative proportions of the different prey taxa, with a progressive decrease with time in the occurrence of krill and a concomitant increase of fish, penguins and squid. Possible influence of the strong 1997/98 ENSO event is discussed.     

Antifouling and antibacterial compounds from a marine fungus <Emphasis Type="Italic">Cladosporium</Emphasis> sp. F14

To investigate the antifouling secondary metabolites from marine-derived fungi, we used bioassay-guided column chromatography techniques, such as HPLC, to separate and purify compounds from Cladosporium sp. F14. Extensive spectral analyses including 1D NMR spectra and MS were employed for structure elucidation of the compounds. Antilarval activity of the compounds was evaluated in settlement inhibition assays with laboratory-reared Balanus amphitrite and Bugula neritina larvae, while antibacterial activity was assessed with disc diffusion bioassay on growth inhibition of six marine bacterial species. In total, nine compounds were obtained. Among them, 3-phenyl-2-propenoic acid, cyclo-(Phe-Pro) and cyclo-(Val-Pro) had various antibacterial activities against three fouling bacteria, furthermore, 3-phenyl-2-propenoic acid and bis(2-ethylhexyl)phthalate effectively inhibited larval settlement of B. neritina and B. amphitrite larvae, respectively, indicating that the two compounds are potential natural antifouling agents.     

Ontogenetic changes in the ecological function of the association behavior between jack mackerel <Emphasis Type="Italic">Trachurus japonicus</Emphasis> and jellyfish

Commensal behavior of jack mackerel Trachurus japonicus (Temminck & Schlegel) with jellyfishes has been widely observed but its ecological function is still unclear. The goal of the present research is to examine the function of association behavior with jellyfish in the laboratory and in field observations with an emphasis on ontogenetic changes. In the laboratory, jack mackerel juveniles (mean standard length (SL) = 11, 19, 38, and 55 mm) were placed in 500-l polycarbonate tanks with two live moon jellyfish, Aurelia aurita (Linné), and one artificial jellyfish made of silicon. Association behavior with either live or artificial jellyfish was visually observed under the following conditions: control, presence of a predator model, before and after feeding live Artemia, 1 h and 3 h after feeding, and at night. Jack mackerel at 11 mm SL associated with both the moon jellyfish and artificial jellyfish, unrelated to the presence of a predator model or feeding. Juveniles at 19 mm associated with moon jellyfish only in the presence of a predator model. Larger juveniles associated with moon jellyfish at 1 h and 3 h after feeding. Thus the ecological function of association was proposed to develop first from school formation, next as a hiding place from predators, and then as a food source. Underwater observations of jack mackerel associating with giant jellyfish Nemopilema nomurai (Kishinouye) in two different areas in the Sea of Japan supported this hypothesis. High predation pressure from benthic piscivorous fishes in the southern area (Tsushima) may encourage association with jellyfish, whereas pressure from pelagic predators in the northern area (Maizuru) may encourage settlement to rocky reef habitats in temperate waters. Thus the jellyfish may also function as a vehicle for the northward migration of this species. Guest editors: K. A. Pitt & J. E. Purcell Jellyfish Blooms: Causes, Consequences, and Recent Advances     

<Emphasis Type="Italic">Bathyphellia margaritacea</Emphasis> (Cnidaria: Actiniaria): the most northern species of the world

The sea anemone Bathyphellia margaritacea (Danielssen in Actinida. The Norwegian North-Atlantic expedition 1876–1878, Groendahl, Oslo, 1890) was collected by the research submersible MIR at the North Pole at a depth of 4,262 m and by the North Pole Drifting Station NP-22 in the American sector of Arctic Ocean covered by permanent ice. These widely separated records significantly increase the known geographic range of the species. B. margaritacea is highly plastic and has an ability to occupy different types of substrates. It appears to be the only species of sea anemone that is able to range in the high Arctic up to the North Pole and the only reliably identified species known from this part of the world.     

A simple molecular approach to distinguish between two Arctic gadoid fishes <Emphasis Type="Italic">Arctogadus glacialis</Emphasis> (Peters, 1874) and <Emphasis Type="Italic">Boreogadus saida</Emphasis> (Lepechin, 1774)

Correct phenotypical identification of and discrimination between Boreogadus saida and Arctogadus glacialis is challenging, especially for larvae and young fish. We propose the use of a single microsatellite genetic marker, Gmo8, to distinguish between the two gadoid fish species. Amplified allele frequencies differed considerably, with B. saida (n = 97) being almost exclusively monomorphic at this locus, whereas A. glacialis (n = 136) is highly polymorphic. There was a clear separation between the amplified allele ranges for the two species. The species specific properties of Gmo8 enables the use of this marker to distinguish between B. saida and A. glacialis.     

High toxicity of the novel bloom-forming species Chattonella ovata (Raphidophyceae) to cultured fish

A toxicological study of an axenic cell line of novel species Chattonella ovata Y. Hara et Chihara (Raphidophyceae) revealed that cultured species of sea bream (Pagrus major), horse mackerel (Trachurus japonicus), and yellowtail (Seriola quinqueradiata) were killed by 4.1–6.8 × 10³, 5.4 × 10³, and 2.8 × 10³ cells/mL, respectively. The sensitivity of the gill lamellae to C. ovata differed among the fish species tested. This finding revealed that C. ovata was highly toxic to the cultured fish. Histological examination showed that edema and hyperplasia of the secondary gill lamellae of red sea bream and horse mackerel occurred when exposed to, or killed by C. ovata, whereas severe damage in the gill lamellae was not observed in yellowtail. Chattonella produced high amounts of superoxide anion radicals and hydrogen peroxide, possibly responsible for the fish death observed. Based on the results of this study and occurrence of a red tide by this organism in China in 2001, we consider this organism to be one of the harmful algae in coastal waters. This is the first report demonstrating that C. ovata is highly toxic to fish, and that it produces superoxide and hydrogen peroxide.     

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.     

Molecular identification of intestinal microflora in Takifugu niphobles

We investigated the intestinal microflora of coastal fish including Takifugu niphobles using both culture techniques and library cloning. As a result, the numbers of bacteria appeared on agar media were 1.0 × 10⁴ to 1.4 × 10⁹ CFU/g (colony forming units/gram), whereas those of total bacteria stained with 4′,6-diamidino-2-phenylindole were 4.7 × 10¹⁰ to 1.9 × 10¹¹ cells/gram, irrespective of different fish species. In addition, the culture technique showed that the intestinal microflora in all specimens was mainly composed of the genus Vibrio. In contrast, the direct count method showed that spirochaetes with length of 2.5-4.5 μm were present in the intestinal contents of T. niphobles at high densities, whereas such bacteria could not be detected in those of other fish species. Library cloning yielded the sequences of 16S rRNA genes that were divided into seven taxonomic categories of bacteria including Actinobacteria, Bacilli, Clostridia, Gammaproteobacteria, Mollicutes, Spirochaetes and an unclassified bacterial group. These results demonstrate that the molecular diversity of the intestinal bacteria in T. niphobles based on the clone library method reflects the direct observation by fluorescence microscopy to some extent.     

Adding coral rubble to substrata enhances settlement of Pocillopora?damicornis larvae

Settlement preferences of Pocillopora damicornis larvae were examined on artificial substrata. Planulation of P. damicornis followed a lunar cycle and the release of larvae occurred after new moon. P. damicornis larvae had the highest rates of settlement within 3 days of being presented settlement substrata. Cumulative settlement gradually increased from 3 to 8 days, and post-settlement mortality was most frequent after 8 days. Settlement experiments showed greatest settlement preference to cement tiles containing 10% coral rubble. This study suggests that physical cues are important in the settlement process, which may be useful for coral reef rehabilitation projects.     

Global population genetic structure of the starlet anemone <Emphasis Type="Italic">Nematostella vectensis</Emphasis>: multiple introductions and implications for conservation policy

Distinguishing natural versus anthropogenic dispersal of organisms is essential for determining the native range of a species and implementing an effective conservation strategy. For cryptogenic species with limited historical records, molecular data can help to identify introductions. Nematostella vectensis is a small, burrowing estuarine sea anemone found in tidally restricted salt marsh pools. This species’ current distribution extends over three coast lines: (i) the Atlantic coast of North America from Nova Scotia to Georgia, (ii) the Pacific coast of North America from Washington to central California, and (iii) the southeast coast of England. The 1996 IUCN Red List designates N. vectensis as “vulnerable” in England. Amplified fragment length polymorphism (AFLP) fingerprinting of 516 individuals from 24 N. vectensis populations throughout its range and mtDNA sequencing of a subsample of these individuals strongly suggest that anthropogenic dispersal has played a significant role in its current distribution. Certain western Atlantic populations of N. vectensis exhibit greater genetic similarity to Pacific populations or English populations than to other western Atlantic populations. At the same time, F-statistics showing high degrees of genetic differentiation between geographically proximate populations support a low likelihood for natural dispersal between salt marshes. Furthermore, the western Atlantic harbors greater genetic diversity than either England or the eastern Pacific. Collectively, these data clearly imply that N. vectensis is native to the Atlantic coast of North America and that populations along the Pacific coast and in England are cases of successful introduction.     

Chemical recognition by hermit crabs of their symbiotic sea anemones and a predatory octopus

The ability of the hermit crabs Dardanus venosus and Pagurus pollicaris to recognize chemically their symbiotic sea anemone Calliactis tricolor and a common octopus predator, Octopus joubini, were studied. The crabs were tested for chemoreceptive responses using a Y-trough olfactometer, which prevented visual cues from being used. Choice tests showed that D. venosus could chemically detect Calliactis. If, however, D. venosus had a Calliactis on its shell at the time of the trial, it could not detect the test anemone. P. pollicaris (without a symbiotic anemone) did not locate the test anemone. Both species of crab avoided a water current carrying octopus chemical cues. Chemoreception may play an important role in the interaction of a hermit crab with its symbiotic anemone and an octopus predator.