Haliotis Rufescens Research Articles

Effect of three bacterial isolates from a commercial hatchery on early red abalone (Haliotis rufescens) postlarvae

Three bacterial strains, GHrC11, GHrC13 and GHrC15, were isolated from an abalone postlarval culture system in a commercial farm at Baja California, Mexico. The strains were phenotypically characterized and sequenced (16S rDNA). Strain GHrC11 was a Gram-positive coccobacillum, while strains GHrC13 and GHrC15 were Gram-negative bacilli. Strain GHrC11 was identified as Exiguobacterium sp. The strains GHrC13 and GHrC15 were identified as Vibrio splendidus. The effects of these strains for the development of early abalone postlarvae (2 days old) were evaluated following a completely randomized design with three replicates using 5-mL-volume Petri dishes as experimental units. The experiment considered two different bacterial concentrations of each strain (103 and 105 cells ml−1) and two controls (with and without the benthic diatom Navicula incerta). After 10 days of experimentation, the highest mortality (90 ± 5.8 %) and the lowest growth rate (4.1 ± 0.1 μm day−1) were recorded for the strain GHrC11. In contrast, the lower mortality (16.7 ± 3.3 %) and the highest growth rate (11.2 ± 0.9 μm day−1) corresponded to the control fed N. incerta. Our results suggest that pathogenic effects of these bacterial strains were stronger than any potential benefits derived from the ingestion of bacteria by early abalone postlarvae. In conclusion, the most pathogenic strain was GHrC11, and the intensity of pathogenicity could be ordered as Exiguobacterium sp. > V. splendidus (C13) > V. splendidus (C15).

Physiological performance of juvenile Haliotis rufescens and Haliotis discus hannai abalone exposed to the withering syndrome agent

Withering syndrome (WS) is a serious chronic disease caused by infection with the bacterium Candidatus Xenohaliotis californiensis, a Rickettsiales-like organism (WS-RLO) that affects multiple abalone species in both natural and farmed populations. However, there is no available information regarding the effects of this disease on the physiological performance of infected abalone. We studied the effect of different levels of infection on components of energy balance and physiological indices (rates of absorption and assimilation, O/N ratio, and scope for growth) in the abalone species Haliotis rufescens and Haliotis discus hannai. Juveniles were exposed to C. X. californiensis transmission for 130days, during which time the presence/absence of WS-RLOs was evaluated by PCR (following DNA sequencing-based confirmation of 100% identity with the sequence of C. X. californiensis from California), and the prevalence and intensity of infection were evaluated via histological analysis. Among H. rufescens juveniles exposed to the bacterium, 92% became infected (positive by histology), and the intensity of infection ranged from low (degree 1) to moderate (degree 2). In contrast, no H. discus hannai juveniles were positive for WS-RLO by histology, although 23% were positive by PCR, possibly indicating incipient WS-RLO infection that did not develop during the experimental period or to mere presence of WS-RLO DNA in the sample. Infection of H. rufescens juveniles by WS-RLOs negatively affected all components of the energy balance and physiological indices, such as scope for growth and the O/N ratio, in direct relation to the degree of infection. The most strongly affected functions were the rate of ingestion, standard metabolism, and production of feces, which were reduced by 60–80% in the most highly infected individuals. The reduced energy intake in the organisms produced a strong energy imbalance such that the energy available for growth was reduced by 49% in infected organisms. In contrast, juveniles of H. discus hannai carrying the bacterium developed no infection and showed no alterations of physiological function. Our results indicate that the level of early infection by WS-RLOs may exert a negative effect on physiological activity in H. rufescens, even when the disease is not evident.

Red abalone collecting and marine water temperature during the Middle Holocene occupation of Santa Cruz Island, California

Oxygen isotope values derived from prehistoric mussel (Mytilus californianus) shell calcite are used to determine whether sea-surface temperatures in the vicinity of the Punta Arena site (CA-SCRI-109) on Santa Cruz Island, California, were cooler than present between 6300 and 5300 cal BP. This site and others in the western sector of the island dating to this period are distinctive because of the presence of large red abalone (Haliotis rufescens) shells, a species that historically has been subtidal around Santa Cruz and the other northern Channel Islands. Comparison of temperature values derived from archaeological shells with those from modern mussel shells, along with water temperatures derived from satellites, indicates that waters were cooler during the 6300 to 5300 cal BP period. These results are consistent with a previous study and support the interpretation that collection of red abalone during the 6300–5300 cal BP period was partly the result of cooler water temperatures that made this large mollusk more available to foragers in the intertidal or shallow subtidal zone.

Mollusk Shell Nacre Ultrastructure Correlates with Environmental Temperature and Pressure

Nacre, or mother-of-pearl, the tough, iridescent biomineral lining the inner side of some mollusk shells, has alternating biogenic aragonite (calcium carbonate, CaCO(3)) tablet layers and organic sheets. Nacre has been common in the shells of mollusks since the Ordovician (450 million years ago) and is abundant and well-preserved in the fossil record, e.g., in ammonites. Therefore, if any measurable physical aspect of the nacre structure was correlated with environmental temperatures, one could obtain a structural paleothermometer of ancient climates. Using X-ray absorption near-edge structure (XANES) spectroscopy, Photoelectron emission spectromicroscopy (PEEM), and X-ray linear dichroism we acquired polarization-dependent imaging contrast (PIC) maps of pristine nacre in cross-section. The new PIC-map data reveal that the nacre ultrastructure (nacre tablet width, thickness, and angle spread) is species-specific in at least eight mollusk species from completely different environments: Nautilus pompilius, Haliotis iris, Haliotis rufescens, Bathymodiolus azoricus, Atrina rigida, Lasmigona complanata, Pinctada margaritifera, and Mytilus californianus. Nacre species-specificity is interpreted as a result of adaptation to diverging environments. We found strong correlation between nacre crystal misorientations and environmental temperature, further supported by secondary ion mass spectrometry measurements of in situ δ(18)O in the nacre of one shell. This has far-reaching implications: nacre texture may be used as a paleothermometer of ancient climate, spanning 450 million years of Earth's history.

Hybridization betweenHaliotis rufescensandHaliotis discus hannai: evaluation of fertilization, larval development, growth and thermal tolerance

Two introduced abalone species are currently produced in Chile, red abalone Haliotis rufescens and Japanese abalone Haliotis discus hannai. However, red abalone accounts for 99% of total production, while the Japanese abalone has not adapted well to Chilean coastal waters. This study reports the hatching, growth and thermal tolerance performance in interspecific hybrids produced between red (R) and Japanese (J) abalone. Our results show that egg age and sperm concentration were critical factors to produce hybrids. The cross R♀ 9 J♂ showed a fertilization rate of 55.3 ± 3.5% using 20-min-old eggs and sperm concentrations of 14 9 10 6 cells mL 1 , while the reciprocal cross (J♀ 9 R♂) was not successful. Further, larval development stages were similar in RR, JJ and RJ hybrid abalones. Among the experimental trials, settlement rate varied from 12.3% to 18.6% and final survival from 20.1% to 31.7%, being the RJ hybrid rates intermediate between parental species. The final shell lengths were similar between RR and RJ hybrids, but significantly higher in JJ abalones. In addition, thermal tolerance was ascertained due its pivotal role for the abalone physiology. Thus, RJ hybrids showed the highest HSP70 gene expression and offers new possibilities to expand Chilean abalone production in warm waters zones.

Boat-based foraging and discontinuous prehistoric red abalone exploitation along the California coast

Temporally and spatially discontinuous pulses of heavy prehistoric exploitation of red abalone (Haliotis rufescens) have been documented along the southern and central California coast. This article examines the very late (post-950cal BP) appearance of numerous red abalone processing sites on the Monterey Peninsula in central California. We test three prominent explanations offered for the sudden onset of red abalone processing sites: trophic cascades resulting from human predation on sea otters, logistical foraging by inland residents, and changes in sea surface temperature. A trophic cascade appears to have occurred but does not fully explain the nature or timing of the phenomenon in the region.We present an alternative explanation that argues that intensive procurement of red abalone emerged at a time when both population pressure and social complexity increased greatly in central California. We argue that a new exploitation strategy—diving from boats—was employed to exploit a much larger portion of the red abalone habitat. This strategy entailed logistical forays by divers who worked new patches in tandem with boaters, gathered large quantities in a single foray, and then field processed them in bulk on the shore before transporting the meat to coastal residences. This strategy provided an additional source of food, and both tradable dried meat and numerous large shells that could be manufactured into ornaments and traded as decorative accoutrements. We conclude our discussion with a consideration of the factors that created such a discontinuous record of red abalone exploitation along the California coast.

Evidence for GABA as an Inhibitory Neurotransmitter in the Neural Circuit Regulating Metamorphosis in the Marine Mollusc Ilyanassa obsoleta

Event Abstract Back to Event Evidence for GABA as an Inhibitory Neurotransmitter in the Neural Circuit Regulating Metamorphosis in the Marine Mollusc Ilyanassa obsoleta Dhani Biscocho1 and Esther M. Leise1* 1 University of North Carolina Greensboro, Biology, United States The marine mud snail, Ilyanassa obsoleta, displays a biphasic life cycle. The initial phase consists of a larval stage in which the animals have ciliated feeding structures on the head (velar lobes) that allow them to swim weakly in the water column. The second phase consists of an adult stage during which the animals reside on intertidal mudflats as obligate omnivores. The event that links these life history stages is metamorphosis. Our interest lies in this transition from larva to non-reproductive adult, the neural circuit that controls metamorphosis, and the cues which are responsible for initiating this process. Molluscan metamorphosis requires multiple changes in cellular action, morphology, physiology and behavioral interactions. I. obsoleta undergoes transformations that include loss of one brain ganglion (the apical ganglion), reorganization of other brain regions, and disappearance of the velar lobes. The neurotransmitter γ-aminobutyric acid (GABA) can induce metamorphosis in a related mollusc, the abalone Haliotis rufescens (Morse, et al. 1979, Science 204, 407-410), but results of our experiments with I. obsoleta have revealed an inhibitory action for GABA. Bath application of GABA to metamorphically competent larvae of I. obsoleta does not elicit metamorphosis. However, exposure of larvae to seawater with reduced chloride concentrations induced metamorphosis in a dose-dependent fashion. Earlier work on I. obsoleta has demonstrated that the neurotransmitters serotonin (5-HT) and nitric oxide function as a promoter and repressor of metamorphosis, respectively. GABA can inhibit serotonergically induced metamorphosis, suggesting that GABA acts either downstream of, or directly on serotonergic neurons. We are currently exploiting reagents that modify the function of GABA transport, synthesis and metabolism to verify that GABA has a role in this system. In bath application, muscimol, a GABA-A agonist, inhibits spontaneous larval metamorphosis for 48 hours. Bath application of isoguvacine, another GABA-A agonist or aminooxyacetic acid (AOA), a major inhibitor of GABA-Transaminase can also repress metamorphosis in 5-HT-induced larvae. In current experiments, we are injecting reagents into competent larvae to decrease chances of interactions between neuroactive compounds and epidermal sensory receptors. These experiments should further support the idea that GABAergic activity occurs internally within the larval central nervous system, perhaps in the apical ganglion. Together, our data will help to elucidate the role of GABA in this molluscan metamorphic pathway. Acknowledgements Funded in part by UNCG Department of Biology Keywords: GABA, gastropod, Larva, metamorphosis, mollusc, Nitric Oxide, Serotonin, Snail Conference: Tenth International Congress of Neuroethology, College Park. Maryland USA, United States, 5 Aug - 10 Aug, 2012. Presentation Type: Poster Presentation (see alternatives below as well) Topic: Development Citation: Biscocho D and Leise EM (2012). Evidence for GABA as an Inhibitory Neurotransmitter in the Neural Circuit Regulating Metamorphosis in the Marine Mollusc Ilyanassa obsoleta. Conference Abstract: Tenth International Congress of Neuroethology. doi: 10.3389/conf.fnbeh.2012.27.00131 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 26 Apr 2012; Published Online: 07 Jul 2012. * Correspondence: Dr. Esther M Leise, University of North Carolina Greensboro, Biology, Greensboro, NC, 27412, United States, esther_leise@uncg.edu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. 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Core structure of aligned chitin fibers within the interlamellar framework extracted from Haliotis rufescens nacre. Part I: Implications for growth and mechanical response

By means of consecutive alkaline and proteolytic treatments of the organic framework?s interlamellar layers extracted from the nacre of H. rufescens, we have exposed a core of aligned parallel chitin fibers. Our findings both verify basic elements of the interlamellar layer structural model of Levi-Kalisman et al. (2001) and extend the more detailed model of Bezares et al. (2008, 2010). We observe via SEM imaging of square millimeter sized samples, which include numerous interlamellar layers and micron sized, yet nanocrystalline, CaCO3 tiles whose native orientation within the shell was first documented, that the chitin fibers in all layers are aligned normal to the growth direction of the shell. Similar alignment has been suggested in the literature for two other classes of mollusks, viz. N. rupertus and P. martensii (Weiner and Traub, 1983; Wada, 1958), suggesting that this may be a more general motif. We find that in order to expose the chitin core it is necessary to first remove protein by an alkaline treatment followed by enzymatic digestion with proteinase-K. We also observe what appear to be the points of traversal of the exposed chitin core by mineral bridges. The implications of these findings touch directly and most specifically upon the expected mechanical properties of organic framework layers such as stiffness and relaxation time constants, viz. they should be planeorthotropic. Single interlamellar layers extracted from nacre should, by implication, also exhibit an orthotropic stiffness. These novel findings provide the structural picture required for a complete anisotropic, time dependent, constitutive description of nacre long thought to be a paradigm of structural optimization. Such a model is briefly described herein and is developed, in full, in Part II of this series.

Multiscale Mechanics and Optimization of Gastropod Shells

A vast majority of mollusks grow a hard shell for protection. The structure of these shells comprises several levels of hierarchy that increase their strength and their resistance to natural threats. This article focuses on nacreous shells, which are composed of two distinct layers. The outer layer is made of calcite, which is a hard but brittle material, and the inner layer is made of nacre, a tough and ductile material. The inner and outer layers are therefore made of materials with distinct structures and properties. In this article, we demonstrate that this system is optimum to defeat attacks from predators. A two-scale modeling and optimization approach was used. At the macroscale, a two-layer finite element model of a seashell was developed to capture shell geometry. At the microscale, a representative volume element of the microstructure of nacre was used to model the elastic modulus of nacre as well as a multiaxial failure criterion, both expressed as function of microstructural parameters. Experiments were also performed on actual shells of red abalone to validate the results obtained from simulations and gain insight into the way the shell fails under sharp perforation. Both optimization and experimental results revealed that the shell displays optimum performance when two modes of failure coincide within the structure. Finally, guidelines for designing two-layer shells were proposed to improve the performance of engineered protective systems undergoing similar structural and loading conditions.

Identification of genes expressed in juvenile Haliotis rufescens in response to different copper concentrations in the north of Chile under controlled conditions

This study reports molecular markers potentially associated with resistance or sensitivity to the impact of copper in juvenile red abalone, Haliotis rufescens, in the north of Chile under experimental conditions. Genomic analysis was made applying subtractive hybridization libraries (SSH) to identify genes up-and down regulated during cooper exposure in abalone over periods of 12 and 168h exposed to 2.5 and 10μg/L of Cu+2. Results obtained from the SSH library revealed 368 different sequences regulated by copper, that correspond to eight major physiological functions. The validation of these sequences obtained by SSH as well as their expression kinetics were made by PCR in real time on 14 potential genes regulated by metal stress. This study provides information for the characterization of potential genomic markers that may be used in future environmental monitoring and to investigate new mechanisms of stress to copper in this commercially important marine species.

Histological evidence of accumulation of iron in postlarvae of red abalone,Haliotis rufescens

The effect of iron on abalone postlarvae (Haliotis rufescens) was investigated in a controlled-culturing system. Three iron concentrations (0.15, 1.5 and 15 mg L−1 of Fe) and a control (no iron added) were used to culture H. rufescens postlarvae while being fed the diatom Navicula inserta over 10 days. Results indicate that H. rufescens postlarvae accumulate iron granules in the stomach, digestive gland and mantle, but not in the gills or other tissues. The number and diameter of iron granules in tissues increased with increasing iron concentration in the culturing environment. The iron accumulation is assumed to have been acquired in the digestive system through the iron-enriched diatom feed and in the mantle through subcutaneous iron transfer. The lack of iron granules in the gills suggests that iron is not absorbed through the respiratory system, as is the case for many filter feeding bivalves. Exposure to the highest iron concentration (15 mg L−1) resulted in tissue abnormalities where granules accumulated, and may have significantly affected the health of H. rufescens postlarvae. These findings provide valuable information for the regulation of appropriate iron levels within aquaculture settings and highlights the importance of monitoring iron levels within abalone larval culturing environments.

Effects of high hydrostatic pressure on microstructure, texture, colour and biochemical changes of red abalone (Haliotis rufecens) during cold storage time

This study was carried out to evaluate the effect of high hydrostatic pressure on quality changes (microstructure, colour, texture and biochemical) of red abalone (Haliotis rufecens) during storage time at 4 °C. High hydrostatic pressure (HHP) treatments were applied at 500 MPa for 8 min and 550 MPa for 3 and 5 min. Biochemical indices covering pH, total volatile basic nitrogen (TVB-N) and trimethylamine (TMA), as well as instrumental texture, microstructure and colour of abalone samples were determined immediately after treatment and throughout subsequent storage at 4 °C. Results have shown that HHP-treated abalones have significantly (p ≤ 0.05) higher pH, moisture and ash content than untreated abalones. Protein and fat contents of treated abalones were significantly (p ≤ 0.05) lower compared to untreated sample (control). TVB-N and TMA levels for HHP-treated abalones rose over the storage period but did not exceed 28 mg TVB-N/100 g and 3 mg TMA/100 g, respectively at the end of 60 days. Instead, the untreated sample exceeded the allowed limit in a 30 day period for the TVB-N and TMA. However, all HHP treatments had less negative effects on tissue colour of abalone than untreated samples in the cold storage time; moreover, whiteness index was reduced to 8% at the end of day 60. A more compact structure was identified as high hydrostatic pressure was higher. Thus, it was concluded that holes in muscle fibres were often due to protein gelation, whenever pressure and protein concentration are high enough, confirming that the structure of abalone muscle treated with high hydrostatic pressure differed significantly from that of raw abalone meat. This paper provides information on microstructural and biochemical stability under prolonged storage time of abalone after pressure treatments which are quite scarce. In base of the data accumulated, criteria for commercial production of high quality abalone with safety requirements could be established.

Sperm chemotaxis, fluid shear, and the evolution of sexual reproduction

Chemical communication is fundamental to sexual reproduction, but how sperm search for and find an egg remains enigmatic. For red abalone (Haliotis rufescens), a large marine snail, the relationship between chemical signaling and fluid motion largely determines fertilization success. Egg-derived attractant plumes are dynamic, changing their size and shape in response to unique combinations of physical and chemical environmental features. Attractant plumes that promote sexual reproduction, however, are limited to a precise set of hydrodynamic conditions. Performance-maximizing shears are those that most closely match flows in native spawning habitats. Under conditions in which reproductive success is chronically limited by sperm availability, gametes are under selection for mechanisms that increase sperm-egg encounter. Here, chemoattraction is found to provide a cheap evolutionary alternative for enhancing egg target size without enlarging cytoplasmic and/or cell volume. Because egg signaling and sperm response may be tuned to meet specific fluid-dynamic constraints, shear could act as a critical selective pressure that drives gamete evolution and determines fitness.

Bioeconomic effect from the size selection in red abalone intensive culture Haliotis rufescens as a production strategy

The variability in growth is a common characteristic in mollusks breeding. Effects rising from the variability in the individual growth rate and the consequent dispersion of sizes in cultivation are important in financial terms. To manage this heterogeneity many firms use size selection, which can happen in two stages: toward the end of the stage of growing, or in the phase of growing of seeds. A bioeconomic model simulating the operation of a firm producing red abalone was implemented in spreadsheets. The firm produces 70 tons yearly. The model was structured in three sub-models. A biological sub-model detailed a batch’s dynamics, in terms of survival and growth, considering individual variation of size around a central value for each age. A technological sub-model described raw materials, the quantity of food and the energy required. Finally, the simulation model is completed with an economic integrated sub-model, where net present value is calculated considering income and costs over the time. Results of the alternative production strategies (with or without selection) are assessed according to: quantity of larvae and necessary spawners to reach the desired level of production; net present value (NPV) and necessary time to recover the investment. The number of larvae was approximately 17 millions larger for the case of the strategy of production with sizes selection and 73% more of available spawners is required for this larger amount of larvae. In the short term, the size-selection strategy increases the production costs at the initial time, compared with the strategy without selection. However, in the long term, this strategy generates greater NPV. The span for investment recovery was shorter for the case of the strategy with size selection and living product (nearly 2,140 days) than frozen (nearly 2,232 days); while without sizes selection a 15-year simulation showed the investment is not recovered. Finally, could be verified that size selection can be an interesting strategy to explore, since it improves the financial result, the same way other more expensive technological changes would.

Relationship between thermal conductivity and structure of nacre from Haliotis fulgens

Abstract

Health and Survival of Red Abalone Haliotis rufescens from San Miguel Island, California, USA, in a Laboratory Simulation of La Niña and El Niño Conditions

The variability in Southern California's marine climate is dominated by the El Niño-Southern Oscillation, with cycling between El Niño events (characterized by warm water, low productivity, and frequent storms) and La Niña events (which exhibit the opposite conditions). Relative to the mainland and other Channel Islands, San Miguel Island (SMI) consistently maintains cooler water temperatures and supports significant populations of red abalone Haliotis rufescens, presumably owing to increased food production and diminished expression of the bacterial disease known as withering syndrome. We conducted a laboratory experiment to examine the effects of La Niña and El Niño conditions on the health and survival of red abalone from SMI. Six replicate tanks per treatment, each containing six abalone, were subjected to one of the following three temperature regimes (treatments): Bodega Bay, California ambient (AMB; mean, 11.4 degrees C), SMI La Niña regime (LAN; mean, 13.8 degrees C), and SMI El Niño regime (ELN; mean, 16.5 degrees C). After 328 d, survival in the ELN treatment was significantly lower than in the AMB and LAN groups. A body condition index was significantly lower in the ELN group than in the AMB group, and the LAN group was in between. A visual score of body shrinkage was significantly higher in both the LAN and ELN groups than in the AMB group. Other clinical signs of withering syndrome and the prevalence and infection intensity of the causative agent, Candidatus Xenohaliotis californiensis, increased as temperature increased in the three treatment groups from AMB to LAN to ELN, although the differences were not statistically significant. Generally, we found that increased temperature resulted in elevated disease expression, although less than observed in previous studies that used farmed abalone and higher, less variable temperature regimes. Water temperature modulates the effect of withering syndrome in wild red abalone, and very strong El Niño events are predicted to result in significant mortality at San Miguel Island.

Reprint of: Growth of nacre in abalone: Seasonal and feeding effects

The processes of aggregation of mineral and organic materials to the growing surfaces in red abalone ( Haliotis rufescens) are analyzed. The flat pearl implantation method is used to observe the transient stages of calcium carbonate deposition, the structure of the organic interlayer, and the steady-state growth of aragonite tiles. The morphology of the organic interlayer is characterized by scanning electron microscopy. These results enable a realistic depiction of the formation of the terraced cones that comprise the principal biomineralization mechanism in this gastropod. In all cases, the growth initiated through spherulites, followed by tile formation. The transient stage with spherulitic formation was shorter at higher temperature; this is indicative of a greater activity of the animal at 21 °C. The growth rate in a normally fed gastropod was found to be higher compared with one provided with limited food. The effect of water temperature (seasonal) was also established, with growth proceeding faster in the summer (T ~ 21 °C) than in winter (15 °C). The structures of the organic interlayer and of the epithelium are revealed by scanning electron microscopy.

Sperm Chemotaxis as Revealed With Live and Synthetic Eggs

Fertilization is one of the least understood fundamental biological processes. How sperm search for and find an egg remains enigmatic. Sperm attraction to egg-derived chemical cues may be significant evolutionarily for maintaining species barriers and important ecologically for increasing gamete encounters. New tools are needed, however, to resolve the functional consequences of these dissolved signal molecules. Freshly spawned eggs from red abalone (Haliotis rufescens) naturally release l-tryptophan, which stimulates chemotactic responses by conspecific sperm. Here, microspheres were manufactured to the approximate size and the same shape as female gametes and formulated to emit controlled doses of chemoattractant, imitating natural l-tryptophan release rates. When experimentally tested for effectiveness, male gametes did not distinguish between chemically impregnated mimics and live eggs, demonstrating that l-tryptophan alone is both necessary and sufficient to promote chemotaxis, and confirming the identity of a native sperm attractant. The techniques that we describe can be used to create synthetic eggs for most animal and plant species, including humans. Egg mimics increase the capacity for experimental manipulation and enable realistic studies of sperm behavior even in the absence of female gametes.

Karyotype analysis of interspecific hybrids between Haliotis rufescens and Haliotis discus HANNAI

Evidence of hybridization in Haliotis has been mainly supported by hatchery experiences and collection of wild hybrid abalones among several species from natural populations worldwide. However, despite the importance to understand the role of the hybridization process through Haliotidae evolution, and also its impact on the abalone aquaculture, genetic studies in hybrid abalones have been poorly developed. Herein, cytogenetic approach allows studying the genetic conformation in hybrid organisms at the chromosome level. This paper reports a quantitative karyotype analysis in Haliotis rufescens, Haliotis discus hannai and their interspecific hybrid. Thus, to characterize chromosome pairs and establish cytogenetic comparisons, chromosome banding with distamycin-A/4,6-diamidino-2-phenylindole fluorochromes and morphologic measurements were performed. The results showed that the hybrids are successfully viable and their karyotypes evidenced a conservative chromosome number of 2n=36. The karyo-idiogram showed a high correspondence in chromosome pair morphology among the hybrids and their parental species, except for a single heteromorphic pair that corresponds to the chromosome 16 from H. rufescens andH. d. hannai respectively. The implications of the abalone hybrid viability derived from its chromosome composition are discussed.

Arrangement of intra-crystalline organic matrix in nacre from red abalone: A view by high-resolution synchrotron powder diffraction combined with SEM observation

In the present study, we combine high-resolution synchrotron powder diffraction with SEM to shed additional light on the arrangement of an intra-crystalline organic matrix in the nacre from red abalone. SEM observation and synchrotron diffraction analysis show that an intra-crystalline organic matrix exists inside nacre tablets and is adsorbed on the (002) atomic plane in an isolated form. Most probably, the intra-crystalline organic matrix is nacrein and its atomic groups are connected with the (012) atomic plane. The findings provide a new understanding of the arrangement of intra-crystalline organic matrices and may facilitate studies on the formation mechanism and the strengthening and toughening mechanism of the nacre. It is worth pointing out that the new experimental method in this present paper can be extended to nanostructural research of other biominerals.