Food resource partitioning by the pufferfishes Sphoeroides spengleri and S. testudineus from Biscayne Bay,Florida

Partitioning of the food resources by two coexisting pufferfishes (Sphoeroides spengleri and S. testudineus) from Biscayne Bay, Florida, USA, was investigated. Gut contents from 453 bandtail and 339 checkered puffers were analyzed. The diets of both species consisted of a variety of benthic prey, but only crustaceans and molluscs were important prey groups. While differences were found in the proportions of general prey categories eaten by these fishes, both species consumed substantial quantities of brachyuran crabs, bivalves, and gastropods. Specific identification of the prey items within these three food categories revealed additional differences in prey between the two puffer species. This partitioning of the food resources by bandtail and checkered puffers was found between both species overall, between overlapping size ranges, and between both species' most abundant size group. Differences in food habits between these two fishes illustrate that congeners with virtually identical mouth structure and complete spatial overlap can significantly partition the food resources.Contribution No. 78-58M from the U.S. Department of Commerce, NOAA, National Marine Fisheries Service, Southeast Fisheries Center, Miami Laboratory, Miami, Florida, USA.     

Occurrence of tetrodotoxin in the gastropodsRapana rapiformis andR. venosa venosa

Paralytic toxicity was detected in four of 17 specimens and six of 28 specimens of the gastropodsRapana rapiformis andR. venosa venosa, respectively, collected from Chiching (Kaohsiung City) and Nanfangao (Ilan County), Taiwan, in 1988 and 1989. The highest toxicities, calculated as tetrodotoxin (TTX) content, were 140 and 13 mouse units (MU) g^–1 digestive gland inR. rapiformis andR. venosa venosa, respectively. The toxins obtained from each species were purified by ultrafiltration and Bio-Gel P-2 column chromatography. The toxin's specific toxicities (as TTX) were 56 MU mg^–1 (R. rapiformis) and 52 MU mg^–1 (R. venosa venosa). Results of analyses by thin-layer chromatography, cellulose acetate membrane electrophoresis and high-performance liquid chromatography showed that TTX and anhydrotetrodotoxin were responsible for the toxicity. The alkali hydrolysate of each toxin showed maximum absorption at ca. 274 nm, which is the unique absorption for the C₉-base of TTX and its related substances (such as anhydrotetrodotoxin, 4-epitetrodotoxin, etc.). Here, we report for the first time the occurrence of TTX in the gastropodsR. rapiformis andR. venosa venosa.     

Electrophoretic studies of serum proteins of four Tilapia species (Pisces)

The blood serum of 4 Tilapia species were analyzed electrophoretically for protein components, and the percentage of proteins in each fraction was determined. Sera from Tilapia nilotica Linnaeus, T. zillii Gervais, T. galilaea Artedi, and T. aurea (Steindachner) were studied. Characteristic protein fractions for each species were obtained. The species differed in the number of protein fractions and in the final mobilities; T. nilotica sera migrate 120 mm, T. zillii sera 95 mm, T. galilaea sera 90 mm, and T. aurea sera 118 mm. The mobilities for fractions 1 and 2 were similar for the 4 species' sera. Inter-specific differences in the amount of protein occurred. Diet may be a major contributing factor to the variations observed in the amount of total protein.     

Accumulation of paralytic shellfish poisoning toxins in planktonic copepods during a bloom of the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis> in Hiroshima Bay,western Japan

Concentrations of paralytic shellfish poisoning (PSP) toxins in toxic dinoflagellate cells and in marine planktonic copepods were monitored during the bloom of Alexandrium tamarense in Hiroshima Bay, western Japan. Concentration of the toxins retained by copepods was a function of the ambient toxin concentration, i.e. the product of A. tamarense cell density and cellular toxicity. The toxin concentration in copepods increased with the increase of toxicants in the seawater then leveled off, but decreased significantly at higher concentrations. In the field, the maximum toxin concentration was 1.2 pmol ind^-1, whereas in the laboratory, the copepod Acartia omorii accumulated a much higher concentration of PSP toxins (24 pmol ind^-1). Feeding avoidance against Alexandrium tamarense and a shift to alternative food sources such as diatoms in the field might keep their toxin levels lower than their potentially maximum level. The copepod toxin levels in the field were not so high as to cause an instantaneous lethal effect on their predator fishes but may reach possibly lethal levels after a few days' continuous feeding. Overall toxin retention by copepods after 12 h feeding and 2 h starvation was only 2.5% of total ingested toxins, which suggested that a significant amount of toxins was released into the seawater. Measurements of toxin reduction and gut evacuation suggested that the toxins were removed through both fecal evacuation and metabolism (e.g. excretion, decomposition and transformation). The results, as a whole, imply that copepods can be a link for PSP toxin flux in both pelagic and benthic food webs and can also be a sink for toxins by metabolizing and removing them from the environment.Communicated by T. Ikeda, Hakodate     

Allelopathic effect of Cylindrospermopsis raciborskii extracts on the germination and growth of several plant species

Cylindrospermopsis raciborskii is an invasive cyanobacterium and a potential producer of the alkaloid toxin cylindrospermopsin (CYN). Extracts of two strains of C. raciborskii were tested for their effects on the germination and growth of Lactuca sativa, Phaseolus vulgaris, Pisum sativum and Solanum lycopersicum. Germination was not significantly inhibited for any of the plant species tested, but growth was affected, depending on the species. Root and stem growth in L. sativa was generally stimulated by both strains. Ph. vulgaris root growth was stimulated by both strains but no effect was visible in stem growth. S. lycopersicum root growth was inhibited by both strains and stem growth was inhibited only by the CYN strain. P. sativum root growth was also inhibited by both strains but stem growth was stimulated. CYN accumulation was also differential with toxin transfer to the stem. Ph. vulgaris accumulated the highest CYN concentration. This study suggests that plants behave differently in their response to this toxin and that roots and stems also show different abilities to react and accumulate the toxin. Knowledge of the impact of CYN- and non-CYN-producing cyanobacteria in different plant species and translocation of the toxin to different plant parts is essential for the avoidance of human as well as environmental health hazards.     

Occurrence of tetrodotoxin in the starfishes Astropecten polyacanthus and A. scoparius in the Seto Inland Sea

Two starfishes, togemomijigai Astropecten polyacanthus and momijigai A. scoparius were collected from the Seto Inland Sea in October 1983 through November 1984, and assayed for toxicity by the standard method for tetrodotoxin (TTX). Most of the 54 A. polyacanthus specimens assayed were toxic, with the highest toxicity score being 520 mouse units (MU) g^-1. All seven A. scoparius were toxic, with the highest score being 46 MU g^-1. The toxin from A. polyacanthus was purified by a method which consisted mainly of activated charcoal treatment, and chromatography on CM-Sephadex C-25 and Bio-Rex 70 columns. The purified starfish toxin showed a specific toxicity of 4 700 MU mg^-1, a value which was almost comparable to that of authentic TTX. From the thin-layer chromatographic and electrophoretic behavior, along with ¹H-NMR spectrum, A. polyacanthus toxin was identified as TTX.     

Effects of resource availability,predators, conspecifics and heterospecifics on decorating behaviour by the majid crab <Emphasis Type="Italic">Tiarinia cornigera</Emphasis>

Utilization of algae for decorating by the intertidal majid crab Tiarinia cornigera was examined by laboratory experiments in relation to availability of algae, presence of a predator, and the presence of conspecific and heterospecific crabs. Different availabilities of decorating materials had a positive correlation on decorating amount by juveniles, while the correlation was not so clear in subadults. The amount of decorating decreased with increasing density of conspecific crabs in the presence of a predator, but there was so clear relationship in the absence of a predator. The decrease in decorating under high density could be due to intraspecific aggression, because the superior crab, in fighting, was found to be decorated with more algae. Tiarinia cornigera was found to be superior to the co-occurring different majid species Micippa platipes in fighting. When T. cornigera and M. platipes were housed together, the former was decorated with more algae than the latter in the presence of a predator, but in the absence of a predator, the amount of algae was not different. Thus, presence of a predator may motivate intraspecific competition in T. cornigera as well as interspecific competition between T. cornigera and M. platipes for decorating materials.     

Toxic dinoflagellates (Alexandrium fundyense and A. catenella) have minimal apparent effects on oyster hemocytes

The possible effect of Alexandrium spp. containing paralytic shellfish poisoning (PSP) toxins on the hemocytes of oysters was tested experimentally. In one trial, eastern oysters, Crassostrea virginica Gmelin, were exposed to bloom concentrations of the sympatric dinoflagellate, Alexandrium fundyense Balech, alone and in a mixture with a non-toxic diatom, Thalassiosira weissflogii (Grun) Fryxell et Hasle. Subsequently, another experiment exposed Pacific oysters, Crassostrea gigas Thunberg, to a mixed suspension of the sympatric, toxic species Alexandrium catenella (Whedon et Kofoid) Balech, with T. weissflogii. Measurements of numbers of oyster hemocytes, percentages of different cell types, and functions (phagocytosis, reactive oxygen species (ROS) production, and mortality) were made using flow-cytometry. During and after exposure, almost no significant effects of Alexandrium spp. upon hemocyte numbers, morphology, or functions were detected, despite observations of adductor-muscle paralysis in C. virginica and measured toxin accumulation in C. gigas. The only significant correlation found was between toxin accumulation at one temperature and higher numbers of circulating live and dead hemocytes in C. gigas. The PSP toxins are known to interfere specifically with sodium-channel function; therefore, the finding that the toxins had no effect on measured hemocyte functions suggests that sodium-channel physiology is not important in these hemocyte functions. Finally, because oysters were exposed to the living algae, not purified toxins, there was no evidence of bioactive compounds other than PSP toxins affecting hemocytes in the two species of Alexandrium studied.     

Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

Changes in microflora of a puffer fish Fugu niphobles,with different water temperatures

Grass puffer Fugu niphobles, captured in November 1986 at Shimoda, Japan, and then reared at different temperatures ranging from 10 to 29°C, was examined for microflora changes in the skin, gill and intestines. At 10°C, the skin and gill were colonized mainly by Pseudomonas, Flavobacterium and Moraxella, while at 29°C Vibrio and Flavobacterium appeared abundantly. The intestinal microflora, consisting of Vibrio, Pseudomonas and Flavobacterium, revealed little temperature dependence, although the intrageneric composition of Vibrio changed conspicuously depending on the change of water temperature. Vibrio group 1, tentatively identified as V. alginolyticus, was detected in all tissues examined at 20 and 29°C. This, along with the fact that V. alginolyticus produces tetrodotoxin, suggests that the tetrodotoxin contained in puffer is, at least partly, accounted for by this bacterial species inhabiting intestines.     

The praying mantis (Mantodea) as predator of the poisonous red-spotted newt <Emphasis Type="Italic">Notophthalmus viridescens</Emphasis> (Amphibia: Urodela: Salamandridae)

A Chinese praying mantis, Tenodera sinensis, was observed feeding on a living red-spotted newt, Notophthalmus viridescens. Specimens of that newt’s population are known to contain high concentrations of tetrodotoxin (TTX), a specific blocker of voltage-gated sodium channels. After experimental oral administration of a TTX-solution (1 mg/ml) to adult specimens of four mantis species, all survived high TTX concentrations (up to 30.8 μg/g body mass) as revealed by analysis of their body extracts, but they are rapidly killed by intra-abdominal injection of 1 μg TTX. The toxin was found to be gradually excreted with faeces. As demonstrated by monoclonal antibody-based immunohistochemical technique, TTX does not penetrate the mid-gut membrane, since it was localized only in the gut lumen, but not in the epithelial cells. This prevents the toxin to reach its target, the sodium channels of the insect’s nervous system, and enables the mantids to feed on toxic prey without risking poisoning.     

Sex pheromone components of pyralid moths Terastia subjectalis and Agathodes ostentalis feeding on the coral tree, Erithrina variegata: Two sympatric species share common components in different ratios

Summary. Two common components, identified as (E)-11-hexadecenal (E11-16:Ald) and (E,E)-10,12-hexadecadienal (E10E12-16:Ald), were found in the extract of virgin females of two sympatric pyralid moths, Terastia subjectalis Lederer, and Agathodes ostentalis (Geyer). The amount of E11-16:Ald and E10E12-16:Ald was 0.12 and 2.8 ng/female for T. subjectalis and 2.0 and 1.8 ng/female for A. ostentalis, respectively. Hexadecanal (16:Ald) was also found at 0.7 ng/female in the extract of A. ostentalis, but EAG activity was unclear. In the field, T. subjectalis and A. ostentalis males were captured with species specific blends in the proportions of 5:95 and 50:50 blends. Blends attractive to one species did not attract the opposite one. Rubber septa loaded with 1 mg each of 5:95 and 50:50 blends of E11-16:Ald and E10E12-16:Ald were as attractive as two virgin females of T. subjectalis and A. ostentalis , respectively. This demonstrated that different ratios of shared pheromone components could provide species specific cues for critical mate location in the two sympatric pyralid moths. It is suggested that directional selection probably occurs in the pheromone blends in opposite directions in both species.     

Immunocytochemical evidence that symbiotic algae secrete potential recognition signal molecules in hospite

Cnidarian–dinoflagellate symbioses are not well understood at the molecular level. Observed specificity between partners during initiation, establishment, and maintenance of the relationship strongly implies a role for chemical signaling. This report presents biochemical and immunocytochemical evidence for potential signaling molecules, as large molecular weight glycoproteins, secreted by Symbiodinium dinoflagellates both in culture and in symbiosis. Polyclonal antibodies directed against recovered exudate from S. microadriaticum, the natural endosymbiont of Cassiopea xamachana, the upside–down jellyfish, were highly specific in recognizing exudates from Symbiodinium species that can successfully induce developmental metamorphosis in the host but did not recognize exudates from Symbiodinium species that do not. Immunoblot analyses showed S. microadriaticum exudate to be protease sensitive. Release of antigenic material by symbiotic S. microadriaticum was demonstrated through light and electron microscopy using immunogold-labeled anti-S. microadriaticum (anti-Sm-XuLg) antibodies as probes. These secreted, symbiont-derived glycoconjugates may be candidates for interspecific molecular signals.     

Predators usurp prey defenses? Toxicokinetics of tetrodotoxin in common garter snakes after consumption of rough-skinned newts

Snakes are common predators of organisms, such as amphibians, with toxic defenses that can be lethal to other predators. Because snakes do not have the option of dissecting prey into edible versus inedible components, they face a full dose of any chemical defenses encountered during attempted predation. This limitation has likely resulted in intense selection favoring the evolution of alternative mechanisms for dealing with prey toxins. These mechanisms can be physiological (e.g., resistance to prey toxins) or behavioral (e.g., toxin sampling and rejection). When physiological resistance arises, the possibility of bioaccumulation of a toxin results. We examined the coevolutionary interaction between the common garter snake (Thamnophis sirtalis) and the rough-skinned newt (Taricha granulosa), which contains a powerful neurotoxin called tetrodotoxin (TTX). In some populations syntopic with newts, individuals of T. sirtalis have evolved resistance to TTX. We examined the persistence of TTX in T. sirtalis after administration of an oral dose of TTX to investigate the possibility that snakes are sequestering TTX. The half-life of TTX in snake liver was estimated at 8.1?days. Accordingly, clearance of 99% of a single dose of TTX averages 61?days. Negative fitness consequences of intoxication during and after newt consumption may be balanced by co-opting the newts?? chemical defense for protection from the snakes?? own predators. Accounting of the coevolutionary dynamic between snakes and newts must incorporate post-consumption affects of lingering TTX.     

Genetic differentiation between populations of Talitrus saltator and Talorchestia deshayesii (Crustacea: Amphipoda) from coastal areas of the north-western European continent

Electrophoretic studies of gene-enzyme variation in the littoral talitrids Talitrus saltator (Montagu) and Talorchestia deshayesii (Audouin) were undertaken to estimate the amount of divergence among geographically separated populations. Samples of both species were taken from sandy beaches over a transect of approximately 3 500 km along the coast of the European continent including Baltic, North Sea and Atlantic locations. A total of 22 T. saltator and 15 T. deshayesii populations were analysed for genetic variation at various enzyme loci. Both amphipods revealed relatively low levels of polymorphism and heterozygosity. Among the loci studied, phosphoglucose isomerase (Pgi) and phosphoglucomutase (Pgm) were highly polymorphic. Patterns of micro- and macrogeographic variation in terms of distributions of allele frequencies at these particular loci are compared. Interpopulation allozymic variation was shown to be lower in T. deshayesii than in T. saltator. As demonstrated by T. saltator populations sampled in coastal sites ranging from Denmark to western France, clinal variation in frequencies of two alleles became evident at the PGI locus, exhibiting a steady increase in the level of polymorphism with decreasing latitudes. It is argued that limited gene flow and, to some extent, random genetic drift may account for the gene pool structure of the talitrid species investigated.     

Chemical defense in pelagic octopus paralarvae: Tetrodotoxin alone does not protect individual paralarvae of the greater blue-ringed octopus (Hapalochlaena lunulata) from common reef predators

Some pelagic marine larvae possess anti-predator chemical defenses. Occasionally, toxic adults imbue their young with their own defensive cocktails. We examined paralarvae of the greater blue-ringed octopus (Hapalochlaena lunulata) for the deadly neurotoxin tetrodotoxin (TTX), and if present, whether TTX conferred protection to individual paralarvae. Paralarvae of H. lunulata possessed 150 ± 17 ng TTX each. These paralarvae appeared distasteful to a variety of fish and stomatopod predators, yet food items spiked with 200 ng TTX were readily consumed by predators. We conclude that TTX alone does not confer individual protection to paralarvae of H. lunulata, and that they possess an alternative defense. In larger doses, tetrodotoxin is a deterrent to the predatory stomatopod Haptosquilla trispinosa (mean dose = 3.97 μg/g). This corresponds to 12–13 paralarvae per predator based on the TTX levels of the clutch we examined. Thus, the basic assumption that individual paralarvae of H. lunulata are defended by TTX alone was disproved. Instead, functionality of TTX levels in paralarvae may arise through alternative selective pathways, such as deterrence to parasites, through kin selection, or against predator species not tested here.     

织纹螺体内的河豚毒素及其衍生物

对2002年福建省引起食物中毒的织纹螺毒素进行研究。高效液相色谱-质谱联用(liquid chromatography/mass,LC/MS)的分析结果显示,织纹螺毒素的主要成分是河豚毒素类毒素(tetrodotoxin,TTX),另外还含有4-epiTTX、anhydroTTX、deoxyTTX等成分。生物法对织纹螺毒素的毒力测定结果表明:所收集的26个织纹螺样品中15个检测出毒性,毒素的毒力在5.7～47.3MU·g-1之间。毒力最高的织纹螺样品2003年5月1日来自福安,主要由半褶织纹螺和另一未定种组成。     

Fate of domoic acid ingested by the copepod <Emphasis Type="Italic">Acartia clausi</Emphasis>

Two important issues in the studies of harmful algae include ecological role of the toxic compounds and their fate through the food web. The aims of this study were to determine whether the production of domoic acid is a strategy evolved to avoid predation and the role of copepods in the fate of this toxic compound through the food web. The copepod Acartia clausi was fed with single and mixed cultures of the toxic diatom Pseudo-nitzschia multiseries and the non-toxic diatom Pseudo-nitzschia delicatissima. Ingestion rate as a function of diatom abundance was the same for the toxic and non-toxic Pseudo-nitzschia species, indicating no selective feeding behaviour against P. multiseries. The toxins ingested by the copepods did not affect mortality, feeding behaviour, egg production and egg hatching of the copepods. Copepods assimilated the 4.8% of the total domoic acid ingested. Although the amount of toxins daily detoxificated by the copepods was 63.6%, the copepods accumulated domoic acid in their tissues. We conclude that domoic acid is not toxic for copepods and, probably for this reason, this toxin does not act as feeding deterrent for copepods. However, even though the production of domoic acid has apparently not evolved to deter predation, copepods may play an important role on the fate of this toxic compound through the marine food web.     

Physiological capacity and environmental tolerance in two sandhopper species with contrasting geographical ranges: Talitrus saltator and Talorchestia ugolinii

We investigated the physiological plasticity and environmental tolerance of two phylogenetically closely related, ecologically similar and co-occurring species of supralittoral amphipods differing drastically in the size of their geographical ranges. A series of physiological traits were characterised for the Corsican-endemic Talorchestia ugolinii Bella-Santini and Ruffo and the widespread Talitrus saltator Montagu. The effect of body mass, temperature and salinity on heart rate (used as proxy for metabolic activity and stress), the effect of temperature on oxygen consumption and the tolerance to salinity exposure were investigated in both species, together with the characterisation of haemolymph osmoregulation in T. ugolinii. Our results showed that there is a clear difference in the resting metabolic rates and physiological capacity, as well as environmental tolerance, between T. saltator and T. ugolinii, with T. saltator overall showing a broader physiological niche. Although T. ugolinii showed a relatively good ability to regulate its haemolymph osmotic concentration (similar to that previously described for T. saltator), it demonstrated a lower tolerance to exposure to hypo-osmotic stress. In addition, a consistent picture emerged between the ability to control the cardiac function and the capacity to actively respond to osmotic stress. The physiological findings are discussed in relation to the known ecology and geographical distribution of T. ugolinii.