Response of hydrothermal vent vestimentiferan <Emphasis Type="Italic">Riftia pachyptila</Emphasis> to differences in habitat chemistry

Vestimentiferan tubeworms, which rely on intracellular sulfide-oxidizing autotrophic bacteria for organic carbon, flourish at deep-sea hydrothermal vents despite the erratic nature of their habitat. To assess the degree to which differences in habitat chemistry (sulfide, pH/CO₂) might impact host and symbiont metabolic activity, Riftia pachyptila tubeworms were collected from habitats with low (H₂S < 0.0001 mM) and high (up to 0.7 mM) sulfide concentrations. The elemental sulfur content of the symbiont-containing trophosome organ was lower in specimens collected from the low-sulfide site. Symbiont abundance, RubisCO activity, and trophosome carbon fixation rates were not significantly different for individuals collected from low- versus high-sulfide habitats. Carbonic anhydrase activities were higher in the anterior gas exchange organs of R. pachyptila from the low-sulfide habitat. Despite large differences in habitat chemistry, symbiont abundance and autotrophic potential were consistent, while the host appears to tailor carbonic anhydrase activity to environmental CO₂ availability.     

Bacterial symbiosis in Northeast Pacific Vestimentifera: a TEM study

A number of new vestimentiferan species occur at northeast Pacific hydrothermal vent sites. The trophosome and bacterial symbionts of three species, collected from the Juan de Fuca and Explorer Ridges between 1984 and 1986, were studied by transmission electron microscopy (TEM). As in Riftia pachyptila, trophosome tissue is organised into lobules each having an axial blood vessel, and intracellular bacterial symbionts are contained in membrane vacuoles. The bacteria have many cytoplasmic inclusions including tubular membrane systems, glycogen-like particles and poly--hydroxybutyrate (PHB) or sulfur bodies. Glycogen production may be quantitatively important to both the symbionts and the host. Glycogen-like granules appear to first accumulate in the bacterial cells and then be released into the bacteriocyte cytoplasm as bacteria are degraded. Although various stages of bacterial growth and degradation are observed, data are insufficient to verify any across-lobule progression of these processes. Morphological comparison of the symbionts reveals that similar symbionts are found in different vestimentiferan species and that one to two bacterial types exist within single individuals. Two possible models of trophosome function and nutrient exchange are discussed.Deceased     

Metabolic and blood characteristics of the hydrothermal vent tube-worm Riftia pachyptila

Specimens of the hydrothermal vent pogonophoran Riftia pachyptila Jones were collected by submersible at a depth of 2 600 m at the 21°N hydrothermal vent site on the East Pacific Rise (20°50N, 109°06W) in April and May of 1982. The worms were maintained in pressurized aquaria for up to 45 d for metabolic studies. Consumption of O₂ was regulated down to low P_{O 2} (oxygen partial pressure) values; O₂ consumption rates were 0.63 and 1.12 mol g^-1 wet wt h^-1 at 2.5° and 8°C, respectively; such rates were comparable to those previously measured for other pogonophorans. Intact specimens of R. pachyptila (including bacterial symbionts) did not consume significant amounts of CH₄ from the environment. The respiratory quotients, in the absence of added sulfide, indicated that metabolism was mainly heterotrophic. High rates of uptake of dissolved amino acids were recorded for one specimen. The total [CO₂] in the vascular blood and the Hb-containing coelomic fluid were high. Under anaerobic conditions, there were equilibrium distributions of pH, total [CO₂] and sulfide concentrations between the vascular blood and the coelomic fluid, apparently because these metabolites were readily exchanged between the two compartments. The vascular blood bound neither CH₄ nor H₂. However, sulfide was reversibly bound by both the vascular blood and coelomic fluid; because this binding depended strongly on pH (with a maximum at about 7.5), HS^- was probably the molecular species bound. Under anaerobic, but not aerobic conditions, the trophosome bound substantial amount of sulfide; thus, the high concentrations of sulfide in the trophosome may have resulted mainly from sulfide bound to sulfide oxidases under anaerobic conditions. The coelomic fluid had a relatively low buffering capacity (2.2 mmol CO₂pH^-1).     

Elemental sulfur in the gills of three species of clams containing chemoautotrophic symbiotic bacteria: a possible inorganic energy storage compound

Sulfur content and fine structure were studied for tissues of three species of clams, Lucinoma annulata, Calyptogena elongata and Lucina floridana, which inhabit sulfide-rich environments and whose gills harbor symbiotic sulfur bacteria. Lucinoma annulata and C. elongata were dredged from the Santa Barbara basin, California, USA, at a depth of 480 to 490 m, and Lucina floridana were dug from below the roots of seagrasses in Saint Joseph Bay, Florida, at a depth of 0.25 to 2m. Foot tissue of Lucinoma annulata, without symbionts, had a total sulfur content of 1.4±0.1 (SD) mg 100 mg^-1 dry weight of tissue (%DW). The symbiont-containing gill tissue of different individuals of L. annulata varied in color from dark red to pale yellow, and the total sulfur content was 2.5±0.4% DW in red gills and was 5.6±3.3 % DW in the yellowest gills. Maintenance of L. annulata in the laboratory for 21 d in the absence of sulfide resulted in the loss from the gill of yellow deposits which were elemental sulfur in the form of liquid-crystalline sulfur globules rather than solid orthorhombic sulfur crystals. The foot tissue did not contain elemental sulfur. When examined by freeze-etch microscopy, sulfur globules were found only within bacteria and not in the animal host cytoplasm. Sulfur globules were confined to the periplasmic space of the bacteria. C. elongata and Lucina floridana resembled Lucinoma annulata in the physical form and distribution of elemental sulfur. The absence of elemental sulfur in the animal cytoplasm suggests that its formation from sulfide is not a detoxification scheme to protect animal tissue from sulfide toxicity. The sulfur deposits probably represent inorganic energy reserves that permit the symbiotic bacteria to function even during the temporary absence of external sulfide.     

The bacterial symbiont from the hydrothermal vent tubewormRiftia pachyptila is a sulfide specialist

We have investigated the metabolic adaptations of the chemolithotrophic bacterial symbionts ofRiftia pachyptila. Specimens of the tubeworm were collected by submersible from depths of 2600 m at 13°N on the East Pacific Rise in 1987, and 2450 m at the Galápagos Rift in 1988. Isolated bacteria utilize sulfide, but not thiosulfate or sulfite, as their sole reduced-sulfur energy source. The bacteria rapidly oxidize a wide range of sulfide concentrations (5µM to 2 mM), with maximal respiration rates at concentrations >1 mM, and unlike many sulfur-oxidizing bacteria, show no inhibition in oxygen consumption at sulfide concentrations up to 2 mM. Incubations of freshly homogenized trophosome tissue or isolated bacteria with sodium [³⁵S] sulfide and subsequent analysis of sulfur products by high-performance liquid chromatography and flow-through scintillation counting showed that sulfide disappeared almost completely within 1 min. Both soluble and insoluble products of sulfide oxidation were produced. The soluble fraction contained sulfate and polysulfides, with no thiosulfate produced. However, the majority of the radioactivity was in the water-insoluble fraction, mostly as elemental sulfur. Whole-worm experiments under pressure showed a rapid removal of³⁵S-sulfide from the incubation water, with sulfide, sulfate, and polysulfides appearing in the blood within 4 h. There was no utilization of thiosulfate by the whole worms, freshly homogenized trophosome tissue, or isolated bacteria.     

An autoradiographic examination of carbon fixation, transfer and utilization in the Riftia pachyptila symbiosis

Riftia pachyptila, the giant vestimentiferan tubeworm from the East Pacific Rise, harbors abundant chemolithoautotrophic, sulfide-oxidizing bacteria in an internal organ, the trophosome. Several facts, such as the lack of a digestive system in the host, stable carbon isotope values and net carbon dioxide uptake all suggest that the tubeworms obtain the bulk of their nutrition from their symbionts. Using tissue autoradiography, we investigated the mode of nutritional transfer between symbionts and host, and the site of early incorporation of symbiont fixed-carbon in the host. Fast labeling in the trophosome clearly demonstrates that the symbionts are the primary site of carbon fixation. Appearance of label in some symbiont-free host tissues in as little as 15 min indicates that the symbionts release a significant amount of organic carbon immediately after fixation. The organic carbon is largely incorporated into specific, metabolically active host tissues such as fast-growing body regions in the trunk and plume, and into tube-secreting glands. In addition to immediate release of fixed carbon by the symbionts, there is evidence of a second possible nutritional mode, digestion of the symbionts, which is consistent with previous suggestions based on trophosome ultrastructure. Results suggest that symbiont-containing host cells migrate in a predictable pattern within trophosome lobules and that symbiont division occurs predominately in the center of a lobule, followed eventually by autolysis/digestion at the periphery of the lobule. Received: 1 July 1999 / Accepted: 30 December 1999     

The enzymes involved in synthesis and utilization of carbamylphosphate in the deep-sea tube worm Riftia pachyptila

 The obligate symbiosis of the deep-sea tube worm Riftia pachyptila with a sulphur-oxidizing bacterium raises important questions concerning its metabolism and metabolic exchanges. In this study, the presence and properties of the enzymes synthesizing and utilizing carbamylphosphate in the arginine and pyrimidine nucleotide pathways were investigated in this worm. The results show that the ammonium-dependent carbamylphosphate synthetase and ornithine transcarbamylase, enzymes involved in the arginine pathway, are present in all body parts of the worm. In contrast, the glutamine-dependent carbamylphosphate synthetase and aspartate transcarbamylase, enzymes involved in the de novo pathway for pyrimidine nucleotides biosynthesis, are present only in the trophosome, the symbiont-harbouring tissue. Although the bacterial nature of these enzymes is not unambigously established, these results strongly suggest that the de novo biosynthesis of pyrimidine nucleotides is limited to the trophosome, the organ where the production of metabolic energy takes place, while the other parts of the worm's body rely on the salvage pathway for the production of the pyrimidine triphosphate nucleotides. Received: 22 June 1998 / Accepted: 26 October 1999     

Astaxanthin deposition in the cuticle of juvenile American lobster (<Emphasis Type="Italic">Homarus americanus</Emphasis>): implications for phenotypic and genotypic coloration

The phenotypic brownish-green color of an American lobster (Homarus americanus Milne-Edwards, 1837) is determined by genetic and dietary mechanisms. Diet influences color through the carotenoid astaxanthin. This pigment can appear red or blue depending if it is bound to protein, and the relative amounts of each form influence lobster color. Carotenoprotein formation was examined in sibling American lobsters rendered white by a diet low in astaxanthin, by feeding a diet containing 0, 55, 110, or 220 g astaxanthin g^–1 for 110 days beginning on 7 February 2004. The relative levels of red and blue in these lobsters were assessed through the analysis of digital photographs of the lobsters, which were taken monthly. The red/blue ratio was used to assess if free (red) or protein-bound astaxanthin (blue) was the dominant form in the lobsters, or if the two forms occurred in ratios allowing for natural coloration. Naturally colored lobsters developed only in the highest astaxanthin diet group, while lobsters fed the 55-g diet were blue. Differences were observed among different parts of the body, and were presumed to be a function of cuticle thickness. However, within each diet treatment, some lobsters initially became red, suggesting an accumulation of free astaxanthin, while other lobsters initially became blue, suggesting an accumulation of protein-bound astaxanthin. This variation may be a result of differences in the uptake of astaxanthin, the rate of carotenoid-protein complexing, or in the total amount of protein available for binding. This variability is likely to be the key to understanding the underlying basis of genetically determined color in American lobsters.Communicated by J.P. Grassle, New Brunswick     

Social behavior and ecology of two sympatric wrasses (Labridae: Halichoeres spp.) off the coast of Florida

The social behavior and ecology of two wrasses common off the coast of Florida (USA), Halichoeres maculipinna and H. garnoti, were studied from late 1975 to mid-1977. Individuals were tagged by means of subcutaneous injection of colored dyes, and the behavior and movements of such tagged fish were noted for as long as 7 months. H. maculipinna appear to be harem-forming, with large and colorful dominant males vigorously defending year-round territories occupied by several subordinate, non-spawning males and several dozen initial-phase fish, most of which are females. The initial-phase fish roam in large, heterospecific feeding aggregates, with each herd remaining in a relatively small joint home range. In contrast, both large colorful and small initial-phase H. garnoti roam as individuals and maintain large, broadly overlapping home ranges. Spawning is apparently promiscuous. Differences in the social systems of the two species appear to reflect differences in their diets and the distribution of food for each on the reef. The behaviors of H. maculipinna and H. garnoti are also compared with those of other studied labrids, with respect to the roles of (1) subordinate males, (2) male-specific color patterns, and (3) factors affecting the success of primary males.     

Activity and inhibitor sensitivity of ATPases in the hydrothermal vent tubeworm Riftia pachyptila : a comparative approach

Phosphorylated ATPases may be involved in the effective pH regulation seen in the hydrothermal vent tubeworm Riftia pachyptila. R. pachyptila appears not only to have a large concentration of ATPases, but the main function of these ATPases seems to have shifted from other types of transport, such as Na⁺ and K⁺ movement, to the facilitation of H⁺ elimination. Plume and trophosome ATPase activity for R. pachyptila measured 646.2 ± 29.5 and 481.4 ± 32.0 μmol Pi (inorganic phosphate) g⁻¹ wet wth⁻¹, respectively. Plume tissue ATPase activity (both mass-specific and protein-specific) in R. pachyptila was higher (between 7% and 55%) than the activity measured in any tissue for 7 other shallow- and deep-living species, in this study. This supports the hypothesis that R. pachyptila regulates acid/base balance via high concentrations of H⁺-ATPases, including Na⁺/H⁺ and K⁺/H⁺ exchangers and possibly electrogenic H⁺-ATPases, as evidenced by a higher total ATPase concentration (646 μmol Pi g⁻¹ wet wt h⁻¹), lesser Na⁺/K⁺-ATPase activity (13% of the total, as compared to 20−40% found in other animals), and higher H⁺-ATPase activity (226–264 μmol Pi g⁻¹wet wt h⁻¹). Overall, R. pachyptila appears to demonstrate elevated ATPase activity, with a greater fraction of the enzymes devoted to proton elimination, in order to effectively control its extracellular pH in the face of processes acting to acidify the internal environment. Received: 9 May 2000 / Accepted: 4 October 2000     

Digestive enzymes in marine invertebrates from hydrothermal vents and other reducing environments

The present study demonstrates the potential hydrolytic activities in the symbiont-containing tissues of the vent invertebrates Riftia pachyptila, Bathymodiolus thermophilus (collected in 1991 at the East Pacific Rise) and the shallow-water bivalve Lucinoma aequizonata (collected in 1991 from the Santa Barbara Basin). Activities of phosphatases, esterases, -glucuronidase and leucineaminopeptidase were comparable to those of digestive tract tissues of other marine invertebrates. A lack in most glycosidases as well as in trypsin and chymotrypsin was observed. Activities of lysozyme and chitobiase were rather high. In all vent invertebrates with symbionts and in L. aequizonata, the symbiont-containing tissues and the symbiont-free tissues had similar levels of enzymatic activities, indicating that polymeric nutrients could be hydrolysed after release from the symbionts and cellular uptake. The high activities of -fucosidase in all vent invertebrates as well as in the shallow-water bivalve L. aequizonata could point to the existence of a yet undescribed substrate available to hydrolysation. The ectosymbionts-carrying polychaete Alvinella pompejana (collected in 1991 at the East Pacific Rise, EPR) shows high lysozyme activities in its gut, consistent with the proposed food source of bacteria. For the vent crab Bythogrea thermydron (also collected in 1991 at the EPR) hydrolytic activities were highest in the gut, dominated by esterase and peptidase activities which support their proposed carnivorous food source. A snail and a limpet collected from R. pachyptila tubes showed high levels of chitobiase suggesting a food source of grazed bacteria or ingested R. pachyptila tube.     

Sulfide-oxidizing bacteria in the burrowing echinoid,Echinocardium cordatum (Echinodermata)

Symbiotic filamentous bacteria thrive in the intestinal caecum of the deposit-feeding echinoid Echinocardium cordatum. Specimens of E. cordatum were collected at Wimereux (Nord Pas-de-Calais, France) in 1991. Their symbiotic bacteria build nodules by forming multilayered mats around detrital particles that enter the caecum. The morphological features of the bacteria are those of Thiothrix, a sulfide-oxidizing genus. The filaments, which may form rosettes, are sheathed and made by a succession of hundreds of rod-shaped bacteria which store elemental sulfur in the presence of external sulfide. Live bacteria are restricted to the outer layers of the nodules. Their sulfide-oxidizing activity was investigated, using a Biological Oxygen Monitor, by measuring the O₂-consumption when reduced sulfur compounds are provided. They oxidize thiosulfate and sulfide. Optimal sulfide oxidation occurs at intermediary pO₂ (100 to 160 M O₂l^-1). Spectrophotometry has confirmed that the sulfur content of the filamentous symbiotic sulfideoxidizing bacteria depends on the presence of external sulfide. This is the first report of symbiotic intradigestive Thiothrixspp.-like bacteria; it lengthens the list of symbioses between sulfide-oxidizing bacteria and invertebrates from sulfide-rich habitats.     

Nuclear and mitochondrial DNA markers indicate unidirectional gene flow of Indo-Pacific to Atlantic bigeye tuna (<Emphasis Type="Italic">Thunnus obesus</Emphasis>) populations,and their admixture off southern Africa

A sharp genetic break separates Atlantic from Indo-Pacific bigeye tuna (Thunnus obesus) populations, as the frequencies of two major mitochondrial (mt) DNA types ( and ) found in this species are different across the tip of southern Africa. The level of nucleotide divergence between mtDNA types and is of the same order as that between reproductively isolated taxa. To further investigate the genetic structure of bigeye tuna over its distribution range and in the contact zone off southern Africa, bigeye tuna samples collected between 1992 and 2001 (including samples from a previous mtDNA survey) were characterized for four nuclear DNA loci and for mtDNA. Nuclear markers did not support the hypothesis that and mitochondria characterize sibling species. Significant allele-frequency differences at one intronic locus (GH2) and one microsatellite locus (µ208) were found between Atlantic and Indo-Pacific samples, although the level of nuclear genetic differentiation (Weir and Cockerhams =0.025 to 0.042) was much lower than in mtDNA ( =0.664 to 0.807). Probabilistic Bayesian assignment of individuals to a population confirmed that southern African bigeye tuna samples represent a simple mixture of individuals from Atlantic and Indian stocks that do not interbreed, with a higher contribution from Indian Ocean individuals (about 2/3 vs. 1/3).Communicated by M.S. Johnson, Crawley     

Population structure of red drum (<Emphasis Type="Italic">Sciaenops ocellatus</Emphasis>) as determined by otolith chemistry

To examine current genetic-based paradigms pertaining to the structure and possible philopatry of red drum populations, we used solution-based inductively coupled plasma-mass spectrometry (ICP-MS) to analyze the otolith chemistry of juvenile red drum (Sciaenops ocellatus) from eight different estuaries in the Gulf of Mexico (Gulf) and the North Atlantic Ocean. One estuary (Tampa Bay, Fla.) was sampled in three different years. Analyses of variance for five elemental ratios (Mg/Ca, Mn/Ca, Zn/Ca, Sr/Ca, Ba/Ca) were all significantly different between estuaries, as was a multi-element signature (MANOVA, Pillais trace F _{50, 1020}=19.41, P<0.0001). We also found that red drum from the Gulf could be distinguished from those taken from the Atlantic Ocean with 99.5% accuracy, likely due to differences in water chemistry between these water masses. A discriminant function developed using these elemental ratios was more than 80% accurate in assigning juvenile red drum to their natal estuary, or in the case of Tampa Bay, to the correct year of spawning. We also used laser ablation ICP-MS to examine the otolith core chemistry of adult red drum collected from spawning aggregations near Tampa Bay. Using a discriminant function analysis with a calibration data set derived from juvenile signatures, we found that 75% of the adult cores matched the juvenile signal established for Tampa Bay 1982. Although preliminary, the results presented here suggest that red drum may return to their natal estuary to spawn, which has been postulated from genetic data.Communicated by P.W. Sammarco, Chauvin     

Reproductive cycle of the endemic cassiduloid<Emphasis Type="Italic"> Cassidulus mitis</Emphasis> (Echinoidea: Cassiduloida) on the Brazilian coast

Cassidulus mitis Krau, 1954 is an endemic species from Rio de Janeiro, Brazil. To study the reproductive cycle of this species, samples were collected at Praia Vermelha, in Guanabara Bay, from October 1998 to April 2000. The sex ratio for C. mitis was 1:1, and reproduction occurred throughout the year. The gonadal index of males was greater than that of females. Five gametogenic stages were identified in males: early growth, premature, mature, partially spawned and early growth with a partially spawned stage. There were no spent or recovery stages such as those seen in females. Females had six gametogenic stages: recovery, early growth, premature, mature, partially spawned and spent stage; there was no early growth with a partially spawned stage as there was in males. The mean oocyte diameter was 382 m (SD=49 m), and the mean number of juveniles per female was 100 individuals (SD=108). The anterior gonads were frequently smaller than the posterior ones, and there were differences in the gametogenic stages between the anterior and posterior gonads. First sexual maturation occurred in individuals with a diameter of 17 mm. The intestinal wet weight was two- to threefold higher than the test wet weight. This may indicate that C. mitis uses its intestine as a weight belt to avoid displacement from the substratum.Communicated by P.W. Sammarco, Chauvin     

Unusual nutrition of the “Pompeii worm” Alvinella pompejana (polychaetous annelid) from a hydrothermal vent environment: SEM,TEM, 13C and 15N evidence

Morphological and histological studies of Alvinella pompejana (a polychaete living in the vicinity of hydrothermal vents of the East Pacific Ocean) were performed using light, scanning and transmission electron microscopy. The worms were collected in April–May 1979, during the Rise cruise by the submersible Alvin on the crest of East Pacific Rise at 21°N. The digestive tracts contained many sulfide particles (as determined by microprobe analysis) associated with organic matter and bacteria. Bacterial communities of different morphological types (cocci and filaments) were also observed at different levels of the worm's outer teguments. An atypical (possibly bacteria-derived) nutritional source of carbon and nitrogen for A. pompejana is indicated by the natural abundances of ¹³C:¹²C and ¹⁵N:¹⁴N in its tissues.Contribution No. 807 from Centre Océanologique de Bretagne, B.P. 337, F-29273 Brest Cédex, France     

The gill symbiont of the hydrothermal vent mussel Bathymodiolus thermophilus is a psychrophilic,chemoautotrophic, sulfur bacterium

Certain hydrothermal vent invertebrates, e.g. Riftia pachyptila and Calyptogena magnifica, are clearly established as harboring dense populations of chemoautotrophic sulfur bacteria in specialized tissues. By contrast, the physiological characteristics of the abundant intracellular gill symbiont of the vent mussel Bathymodiolus thermophilus have been questioned. The low activities of enzymes diagnostic for CO₂ fixation (Calvin cycle) and for sulfur-driven energy generation, as measured by other investigators, have been attributed to bacterial contamination of the gill surface. Based on research at the Galápagos Rift hydrothermal vents in 1988 and subsequent laboratory experiments, the current study confirms that the B. thermophilus symbiont is a psychrophile for which thiosulfate and sulfide stimulate CO₂ fixation. It strongly indicates that the symbiont is a chemoautotroph by establishing the following: (1) Sulfide and thiosulfate can stimulate CO₂ fixation by partially purified symbionts by up to 43-fold and 120-fold, respectively; (2) the ribulose-1,5-bisphosphate carboxylase/oxygenase activity of the symbiont is sufficient to account for its sulfide- or thiosulfate-stimulated CO₂ incorporation; (3) the symbiont's molar growth yield on thiosulfate, as judged by CO₂ incorporation, is indistinguishable from that of free-living chemoautotrophs. Due to the high protein-degrading activity of B. thermophilus gill lysate, it is also suggested that host lysis of symbionts plays a more important role in the nutrition of the vent mussel than in R. pachyptila or C. magnifica, for which no comparable protein-degrading activity was found.     

Ground gleaning in horseshoe bats: comparative evidence from<Emphasis Type="Italic"> Rhinolophus blasii</Emphasis>,<Emphasis Type="Italic"> R. euryale</Emphasis> and<Emphasis Type="Italic"> R. mehelyi</Emphasis>

The 71 species of horseshoe bat (genus Rhinolophus) use echolocation calls with long constant-frequency (CF) components to detect and localize fluttering insects which they seize in aerial captures or glean from foliage. Here we describe ground-gleaning as an additional prey-capture strategy for horseshoe bats. This study presents the first record and experimental evidence for ground-gleaning in the little-studied Blasius horseshoe bat (Rhinolophus blasii). The gleaning bouts in a flight tent included landing, quadrupedal walking and take-off from the ground. The bats emitted echolocation calls continuously during all phases of prey capture. Both spontaneously and in a choice experiment, all six individuals attacked only fluttering insects and never motionless prey. These data suggest that R. blasii performs ground-gleaning largely by relying on the same prey-detection strategy and echolocation behaviour that it and other horseshoe bats use for aerial hawking.We also studied the Mediterranean horseshoe bat (R. euryale) in the flight tent. All four individuals never gleaned prey from the ground, though they appeared to be well able to detect fluttering moths on the ground. It is not known yet whether ground-gleaning plays a role in Mehelys horseshoe bat (R. mehelyi). In a performance test, we measured the ability of these three European species of middle-sized horseshoe bats (R. euryale, R. mehelyi and R. blasii) to take-off from the ground. All were able to take flight even in a confined space; i.e. the willingness to ground-glean in R. blasii is not related to a superior take-off performance. In contrast to ground-gleaning bats of other phylogenetic lineages, R. blasii appears not to be a specialist, but rather shows a remarkable behavioural flexibility in prey-capture strategies and abilities. We suggest that the key innovation of CF echolocation paired with behavioural flexibility in foraging strategies might explain the evolutionary success of Rhinolophus as the second largest genus of bat.Communicated by T. Czeschlik     

Trophic variation with length in two ommastrephid squids, <Emphasis Type="Italic">Ommastrephes bartramii</Emphasis> and <Emphasis Type="Italic">Sthenoteuthis oualaniensis</Emphasis>

From 1998 to 2001 a total of 200 Ommastrephes bartramii (27 paralarvae) and 170 Sthenoteuthis oualaniensis (14 paralarvae) were sampled from the Central North Pacific. One group of non-paralarval O. bartramii (n = 30) was sampled from farther northwest in 1996. The δ¹⁵N of mantle muscle of non-paralarval O. bartramii ( = 12.4‰) was significantly greater than that of non-parlarval S. oualaniensis ( = 8.1‰) (P < 0.001). The δ¹⁵N of whole paralarvae of O. bartramii ( = 6.4‰) was not significantly different than parlarvalae of S. oualaniensis ( = 6.1‰) (P = 0.528). There was no significant difference between the mantle muscle δ¹⁵N values of male (n = 95, = 13.3‰) and female (n = 18, = 12.9 ‰) O. bartramii greater than 300 mm mantle length (ML) (P = 0.15). There was also no significant difference between the mantle muscle δ¹⁵N values of male (n = 15, = 7.2‰) and female (n = 26, = 7.3 ‰) S. oualaniensis in the same size range (P = 0.41). Overall there was a distinct logistic increase in δ¹⁵N with mantle length for O. bartramii, whereas S. oualaniensis showed an exponential increase in δ¹⁵N with mantle length that was stronger within individual years than with all samples combined. In general, adult O. bartramii are more than a trophic level above S. oualaniensis (4.3‰, 1.3 TLs). Because of the nature of the sampling protocol, this study could not separate spatial and temporal effects on the δ¹⁵N signals from each squid species. This study demonstrates the ability of stable isotope analyses to differentiate trophic levels between squid species as well as track trophic changes across size ranges from paralarvae to adults. Additional research is needed to validate these trophic changes across size within individuals.