Marine Snail Research Articles

Mitogenomic phylogeny of Nassarius (Gastropoda: Neogastropoda)

AbstractNassariids (Family Nassariidae) are a group of marine snails that are distributed worldwide, with their maximum species diversity in tropical regions, particularly the Indo‐Pacific. However, the traditional taxonomy of Nassariidae defined by shell or radula characters is usually inconsistent with little phylogenetic signal. In the present study, the complete mitochondrial (mt) genomes of nine Nassarius species were sequenced and compared with other eight nassariid species previously reported. All nassariid mt genomes showed the same gene order as in most caenogastropods and shared a very similar pattern with respect to genome size, nucleotide composition and AT contents. A deletion of three nucleotides in nad6 gene was detected in Nassarius jacksonianus and Nassarius acuticostus, and this feature also provided implications for nassariid phylogeny. The genetic distance analysis and reconstructed phylogeny revealed a distant relationship between N. jacksonianus or N. acuticostus and other members in Nassarius. The mitogenomic phylogeny recovered the evolutionary relationships within Nassarius with high statistical support. In addition, a chronogram was reconstructed under an uncorrelated relaxed molecular clock, which dated the divergence among main lineages of Nassarius during ~31 MYA.

Sample checklist of Gastropoda and Bivalvia in Cham Islands, Vietnam.

BackgroundCham Islands (Cu Lao Cham) is a group of 8 small islands in the Quang Nam province, Central Vietnam. There is only one study that mentioned the diversity of marine molluscs in this area. However, the data on species composition have not been digitised and not stored or shared for other purposes. Our paper aims to share the checklist of marine mollusc species (Bivalvia and Gastropoda) species that we collected from the littoral zone of Cham Islands in May 2017. This is the first digitised and online data of marine molluscs in Vietnam. It is very important for researchers in various fields such as the structure and function of ecology and biodiversity monitoring.New informationThis study provides a checklist of the marine bivalves and snails in the Cham Islands of Vietnam. Moreover, this first widely shared data of biodiversity in Vietnam can trigger biodiversity data papers in this data-poor country.The data of this study will be important inputs for better understanding biodiversity on the Cham Islands and Vietnam as well as for forming the basis for monitoring, exploitation and conservation of biodiversity in this area.In total, 145 taxa were recorded, 46 bivalve taxa and 99 snail taxa, from which 128 were identified to the species level and 17 were identified to the genus level. There are 116 new species records for the Cham Islands.The specimens are currently deposited in the Institute of Ecology and Biological Resources (IEBR), Vietnam Academy of Science and Technology (VAST).

Mutagenesis of α-Conotoxins for Enhancing Activity and Selectivity for Nicotinic Acetylcholine Receptors.

Nicotinic acetylcholine receptors (nAChRs) are found throughout the mammalian body and have been studied extensively because of their implication in a myriad of diseases. α-Conotoxins (α-CTxs) are peptide neurotoxins found in the venom of marine snails of genus Conus. α-CTxs are potent and selective antagonists for a variety of nAChR isoforms. Over the past 40 years, α-CTxs have proven to be valuable molecular probes capable of differentiating between closely related nAChR subtypes and have contributed greatly to understanding the physiological role of nAChRs in the mammalian nervous system. Here, we review the amino acid composition and structure of several α-CTxs that selectively target nAChR isoforms and explore strategies and outcomes for introducing mutations in native α-CTxs to direct selectivity and enhance binding affinity for specific nAChRs. This review will focus on structure-activity relationship studies involving native α-CTxs that have been rationally mutated and molecular interactions that underlie binding between ligand and nAChR isoform.

Short-term exposure to tricyclic antidepressants delays righting time in marine and freshwater snails with evidence for low-dose stimulation of righting speed by imipramine

Active pharmaceutical ingredients such as tricyclic antidepressants (TCAs) are contaminants of emerging concern which are commonly detected in wastewater effluent and which can disrupt the behavior of non-target organisms. In aquatic snails, the righting response is a critical behavior that has been shown to be inhibited by exposure to SSRI-type antidepressants. We exposed marine and freshwater snails to three tricyclic antidepressants (clomipramine, amitriptyline, and imipramine) for 1h and measured righting response time. In the marine mud snail (Ilyanassa obsoleta), all three TCAs significantly increased righting time at concentrations as low as 156μg/L. Similarly, in the freshwater snail Leptoxis carinata, all three TCAs increased righting time at concentrations as low as 263μg/L. However, exposure to imipramine from 15.8 to 316μg/L resulted in significantly faster righting time. Such low-dose stimulation and high-dose inhibition are characteristics of a hormetic response. We discuss the possible physiological mechanism of action of TCAs and other antidepressants on snail behavior, and the occurrence of non-monotonic, hormetic dose responses to human pharmaceuticals in the aquatic environment.

First use of a non-invasive technique for determination of sex hormones in the queen conch Lobatus gigas, Mollusca Gastropoda

Profiles were generated of the sex hormones estrogen, progesterone, and testosterone in the marine snail Lobatus gigas every 2 months for 1 year. A non-invasive technique involving feces collection was used. Hormones were extracted from the feces with 80% methanol. After filtering, hormone concentrations were measured by high-performance liquid chromatography and a UV detector. All three sex hormones were present in L. gigas feces, and their concentrations increased contemporaneously with the conch reproductive period. Males and females showed the highest concentration of testosterone in May (1.8 ± 0.3 and 2.1 ± 0.8 ng/ml, respectively). Both sexes presented a maximum estrogen concentration in May (1.2 ± 0.7 and 1.0 ± 0.3 ng/ml). Progesterone in females remained constant from March to July. Pearson correlation between estrogen and spawning activity were r = 0.66 (p = 0.03) and for testosterone (r = 0.5216) and progesterone (r = 0.437). This study constitutes the first use of this technique in this species. Results show that sex hormones may be controlling this species’ reproductive events, as occurs in other gastropods. The understanding of the L. gigas reproductive process, this is one of several steps that will allow in the future improve aquaculture systems and supporting conservation of wild populations.

Conopeptides [V11L;V16D]ArIB and RgIA4: Powerful Tools for the Identification of Novel Nicotinic Acetylcholine Receptors in Monocytes.

Venomous marine snails of the genus Conus employ small peptides to capture prey, mainly osteichthyes, mollusks, and worms. A subset of these peptides known as α-conotoxins, are antagonists of nicotinic acetylcholine receptors (nAChRs). These disulfide-rich peptides provide a large number of evolutionarily refined templates that can be used to develop conopeptides that are highly selective for the various nAChR subtypes. Two such conopeptides, namely [V11L;V16D]ArIB and RgIA4, have been engineered to selectively target mammalian α7∗ and α9∗ nAChRs, respectively, and have been used to study the functional roles of these subtypes in immune cells. Unlike in neurons and cochlear hair cells, where α7∗ and α9∗ nAChRs, respectively, function as ligand-gated ion channels, in immune cells ligand-evoked ion currents have not been demonstrated. Instead, different metabotropic functions of α7∗ and α9∗ nAChRs have been described in monocytic cells including the inhibition of ATP-induced ion currents, inflammasome activation, and interleukin-1β (IL-1β) release. In addition to conventional nAChR agonists, diverse compounds containing a phosphocholine group inhibit monocytic IL-1β release and include dipalmitoyl phosphatidylcholine, palmitoyl lysophosphatidylcholine, glycerophosphocholine, phosphocholine, phosphocholine-decorated lipooligosaccharides from Haemophilus influenzae, synthetic phosphocholine-modified bovine serum albumin, and the phosphocholine-binding C-reactive protein. In monocytic cells, the effects of [V11L;V16D]ArIB and RgIA4 suggested that activation of nAChRs containing α9, α7, and/or α10 subunits inhibits ATP-induced IL-1β release. These results have been corroborated utilizing gene-deficient mice and small interfering RNA. Targeted re-engineering of native α-conotoxins has resulted in excellent tools for nAChR research as well as potential therapeutics. ∗indicates possible presence of additional subunits.

Investigation of Antibacterial Activity of Crude Extracts from Marine Snails and Bivalves in the Southern Coast of Vietnam

The primary antibacterial activity of methanol and chloroform crude extracts from marine snails and bivalves was assessed by using the agar diffusion technique against four bacterial strains. Active methanol extracts were then characterized using TLC, SDS-PAGE and FTIR. Methanol extracts from 5 snail species and 8 extracts from 12 bivalve species possessed the ability to inhibit <i>Bacillus subtilis</i>. Methanol extracts from 3 snail species <i>Tectus conus, Maninella alounia</i> and <i>Trochus maculatus</i> inhibited <i>Escheria coli</i> and those from 4 snail species <i>Cerithium chinatum, Maninella alounia, Tectus pyramis, Trochus maculatus</i> and the bivalve species <i>Pinna bicolor</i> exhibited activity against <i>Serratia marcescens</i>. Chloroform extracts from 7 snail species and those from 7 bivalve species showed inhibition on <i>Bacillus subtilis</i>. Only chloroform extract from the bivalve <i>Chama cf dunkeri</i> was active on <i>Salmonella typhimur</i> and that from the snail <i>Trochus maculatus</i> and bivalve <i>Lopha cristagali</i> inhibited <i>Escheria coli</i>. TLC and FTIR analysis of active methanol extracts showed the presence of amino acids, peptides and proteins. SDS-PAGE of those extracts also revealed proteins with a molecular weight range between 10 and 28 kDa. The obtained results indicate the potential antimicrobial compounds that could be explored in snail and bivalve in Vietnam.

Sex Determination, Sexual Development, and Sex Change in Slipper Snails.

Sex determination and sexual development are highly diverse and controlled by mechanisms that are extremely labile. While dioecy (separate male and female functions) is the norm for most animals, hermaphroditism (both male and female functions within a single body) is phylogenetically widespread. Much of our current understanding of sexual development comes from a small number of model systems, limiting our ability to make broader conclusions about the evolution of sexual diversity. We present the calyptraeid gastropods as a model for the study of the evolution of sex determination in a sequentially hermaphroditic system. Calyptraeid gastropods, a group of sedentary, filter-feeding marine snails, are sequential hermaphrodites that change sex from male to female during their life span (protandry). This transition includes resorption of the penis and the elaboration of female genitalia, in addition to shifting from production of spermatocytes to oocytes. This transition is typically under environmental control and frequently mediated by social interactions. Males in contact with females delay sex change to transition at larger sizes, while isolated males transition more rapidly andat smaller sizes. This phenomenon has been known for over a century; however, the mechanisms that control the switch from male to female are poorly understood. We review here our current understanding of sexual development and sex determination in the calyptraeid gastropods and other molluscs, highlighting our current understanding of factors implicated in the timing of sex change and the potential mechanisms. We also consider the embryonic origins and earliest expression of the germ line and the effects of environmental contaminants on sexual development.

Identification of Conotoxins with Novel Odd Number of Cysteine Residues from the Venom of a Marine Predatory Gastropod Conus leopardus Found in Andaman Sea

Conotoxins are neuro-pharmacologically active cysteine rich peptides isolated from the venom complex of marine cone snails. These are usually made of even number of cysteines. In this study we characterised six novel conotoxin sequences from the venom of Conus leopardus collected from the Andaman Sea, namely Le907 (C-C), Le868 (C-C), Le933 (-C-CC), Le949 (-C-CC), Le1988 (C-C-CC-C) and Le1642 (CC-C-C) using de novo mass spectrometrybased sequencing methods. Astonishingly 3 of these peptides possess novel arrangements of cysteine residues with odd number of cysteines (-C-CC; C-C-CC-C), namely Le933, Le949 and Le1988. Further, a post-translational variant of peptide Le933 was identified and experimentally determined to contain hydroxyproline. The unusual cysteine arrangements observed suggests novel class of conotoxins. These results expand our understanding of the diversity of odd cysteine arrangements in conotoxins.

Cone snail prolyl-4-hydroxylase α-subunit sequences derived from transcriptomic data and mass spectrometric analysis of variable proline hydroxylation in C. amadis venom

Putative prolyl-4-hydroxylase (P4H) α-subunit sequences have been extracted by mining transcriptomic data obtained from seven cone snail species C. amadis, C. monile, C. araneosus, C. miles, C. litteratus, C. frigidus, and C. ebraeus. Sequences ranging from 518 to 559 residues have been compared with representative animal P4H sequences. The α-subunit consists of an N-terminus double domain, involved in dimerization and substrate binding, while the C-terminus contains the catalytic domain. Definitive functional annotation of the cone snail sequences has been achieved by an analysis of conserved residues responsible for catalytic function, specific conformational features, and subunit interactions, using two independent structures of the double domain, and the catalytic domain, previously reported in the literature. The variability of proline hydroxylation in conotoxins is illustrated by a mass spectrometric analysis of C. amadis venom. Site specific hydroxylation and the presence of peptides with multiple proline residues, resistant to modification, suggests that sequence and conformational effects may determine the substrate specificity of the Conus prolyl-4-hydroxylases. SignificanceProline hydroxylation is a widely observed post translational modification, with collagen being the pre-eminent example. Hydroxylation of proline is also widely observed in conotoxins, which are a major component of marine cone snail venom. This paper describes newly identified prolyl-4-hydroxylase sequences, using transcriptome data from seven Conus species. The predicted functional annotation of prolyl-4-hydroxylase sequences was carried out using two available crystal structures of independent domains. The mass spectrometric characterisation of proline/hydroxyproline containing peptides in C. amadis venom confirms sequence specific hydroxylation in Conus venom as shown previously by others.

Conus Envenomation of Humans: In Fact and Fiction.

Prominent hallmarks of the widely distributed, mainly tropical marine snail genus Conus are: (1) its unusually high species diversity; it is the largest genus of animals in the sea, with more than 800 recognized species; and (2) its specialized feeding behavior of overcoming prey by injection with potent neurotoxic, paralytic venoms, and swallowing the victim whole. Including the first report of a human fatality from a Conus sting nearly 350 years ago, at least 141 human envenomations have been recorded, of which 36 were fatal. Most Conus species are quite specialized predators that can be classified in one of three major feeding guilds: they prey exclusively or nearly so on worms, primarily polychaete annelids, other gastropods, sometimes including other Conus species, or fishes. These differences are shown to relate to the severity of human envenomations, with the danger increasing generally in the order listed above and a strong likelihood that all of the known human fatalities may be attributable solely to the single piscivorous species C. geographus.

Discovery of Novel Conotoxin Candidates Using Machine Learning.

Cone snails (genus Conus) are venomous marine snails that inject prey with a lethal cocktail of conotoxins, small, secreted, and cysteine-rich peptides. Given the diversity and often high affinity for their molecular targets, consisting of ion channels, receptors or transporters, many conotoxins have become invaluable pharmacological probes, drug leads, and therapeutics. Transcriptome sequencing of Conus venom glands followed by de novo assembly and homology-based toxin identification and annotation is currently the state-of-the-art for discovery of new conotoxins. However, homology-based search techniques, by definition, can only detect novel toxins that are homologous to previously reported conotoxins. To overcome these obstacles for discovery, we have created ConusPipe, a machine learning tool that utilizes prominent chemical characters of conotoxins to predict whether a certain transcript in a Conus transcriptome, which has no otherwise detectable homologs in current reference databases, is a putative conotoxin. By using ConusPipe on RNASeq data of 10 species, we report 5148 new putative conotoxin transcripts that have no homologues in current reference databases. 896 of these were identified by at least three out of four models used. These data significantly expand current publicly available conotoxin datasets and our approach provides a new computational avenue for the discovery of novel toxin families.

Two Novel Multi-Functional Peptides from Meat and Visceral Mass of Marine Snail Neptunea arthritica cumingii and Their Activities In Vitro and In Vivo.

Neptunea arthritica cumingii (Nac) is a marine snail with high nutritional and commercial value; however, little is known about its active peptides. In this study, two multi-functional peptides, YSQLENEFDR (Tyr-Ser-Gln-Leu-Glu-Asn-Glu-Phe-Asp-Arg) and YIAEDAER (Tyr-Ile-Ala-Glu-Asp-Ala-Glu-Arg), were isolated and purified from meat and visceral mass extracts of Nac using a multi-bioassay-guided method and were characterized by using liquid chromatography-tandem mass spectrometry. Both peptides showed high antioxidant, angiotensin-converting enzyme (ACE)-inhibitory, and anti-diabetic activities, with half-maximal effective concentrations values less than 1 mM. Antioxidant and ACE-inhibitory activities were significantly higher for YSQLENEFDR than for YIAEDAER. In a zebrafish model, the two peptides exhibited strong scavenging ability for reactive oxygen species and effectively protected skin cells against oxidative damage without toxicity. Molecular docking simulation further predicted the interactions of the two peptides and ACE. Stability analysis study indicated that the two synthetic peptides maintained their activities under thermal stress and simulated gastrointestinal digestion conditions. The low molecular weight, high proportion of hydrophobic and negatively-charged amino acids, and specific C-terminal and N-terminal amino acids may contribute to the observed bio-activities of these two peptides with potential application for the prevention of chronic noncommunicable diseases.

Using Drosophila behavioral assays to characterize terebrid venom-peptide bioactivity

The number of newly discovered peptides from the transcriptomes and proteomes of animal venom arsenals is rapidly increasing, resulting in an abundance of uncharacterized peptides. There is a pressing need for a systematic, cost effective, and scalable approach to identify physiological effects of venom peptides. To address this discovery-to-function gap, we developed a sequence driven:activity-based hybrid approach for screening venom peptides that is amenable to large-venom peptide libraries with minimal amounts of peptide. Using this approach, we characterized the physiological and behavioral phenotypes of two peptides from the venom of predatory terebrid marine snails, teretoxins Tv1 from Terebra variegata and Tsu1.1 from Terebra subulata. Our results indicate that Tv1 and Tsu1.1 have distinct bioactivity. Tv1 (100 µM) had an antinociceptive effect in adult Drosophila using a thermal nociception assay to measure heat avoidance. Alternatively, Tsu1.1 (100 µM) increased food intake. These findings describe the first functional bioactivity of terebrid venom peptides in relation to pain and diet and indicate that Tv1 and Tsu1.1 may, respectively, act as antinociceptive and orexigenic agents. Tv1 and Tsu1.1 are distinct from previously identified venom peptides, expanding the toolkit of peptides that can potentially be used to investigate the physiological mechanisms of pain and diet.

Identifying novel conopepetides from the venom ducts of Conus litteratus through integrating transcriptomics and proteomics

The venom ducts of marine cone snails secrete highly complex mixtures of cysteine-rich active peptides, which are generally known as conotoxins or conopeptides and provide a potential fertile resource for pharmacological neuroscience research and the discovery of new drugs. Previous studies have devoted substantial effort to the identification of novel conopeptides, and the 109 cone snail species have yielded 7000 known conopeptides to date. Here, we used de novo deep transcriptome sequencing analyses combined with traditional Sanger sequencing and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) to identify 30 distinct conopeptide precursors. Twenty of these were previously reported and the other 10 were novel conopeptide precursors. The study provides the first identification of the Con-ikot-ikot, NSF-bt05, O3 and I1 gene superfamilies in C. litteratus. A new putative superfamily was identified. In addition, the following cysteine frameworks were first identified in this study: CC-C-C-C-C-C-C-C-C-C-C-C-CC-C-C-C-C-C and C-C-C-C-C-CC-C. Several isomerases involved in post-translational modification of conopeptides were identified as well. The discovery of new conopeptides in C. litteratus will enhance our understanding of the conopeptide diversity in this particular clade of cone snails. We also found the existence of intraspecific variations in vermivorous species. Finally, the analysis strategy offers a relatively reliable workflow for screening for peptide drug candidates. SignificanceThese novel conopeptides provide a potential resource for the development of new channel-targeting drugs.The intraspecific variation in C. litteratus enhance our understanding of the conopeptide diversity in this particular clade of cone snails.The identified three cysteine residues, which might participate in the formation of disulfide bonds, provide a clue to get the connectivity of cysteine frameworks.Finally, the analysis strategy offers a relatively reliable workflow for screening for peptide drug candidates.

The Full Hierarchical Structuration of Species Abundances Reliably Inferred from the Numerical Extrapolation of Still Partial Samplings: A Case Study with Marine Snail Communities in Mannar Gulf (India)

The detailed analysis of Species Abundance Distributions (S.A.D.s) can shed light on how member-species organize themselves within communities, provided that the complete distribution of species abundances is made available first. In this perspective, the numerical extrapolation applied to incomplete S.A.D. can effectively compensate for incompleteness, when having to deal with substantially incomplete samplings. Indeed, almost as much information can be released from extrapolated S.A.D.s as would be obtained from truly complete S.A.D.s, although the taxonomic identities of unrecorded species remain of course ignored by numerical extrapolation. To take full advantage of this new approach, a recently developed procedure allowing the least-biased numerical extrapolation of S.A.D.s has been applied to three partially sampled gastropods communities associated to coral-reef in Mannar Gulf (S-E India). The following main results were derived from the three numerically completed S.A.D.s: (i) once completed, all three numerically S.A.D.s fairly well comply with the log-normal model, suggesting that, in all three communities, the process of hierarchical structuration of species abundances likely involves the combined contributions of many independent factors; (ii) moreover, the same holds true for each of the two coexisting feeding guilds (primary and secondary consumers) considered separately; (iii) when compared to secondary consumers, the guild of primary consumers has a much lower species richness but, on the other hand, exhibits a more strongly structured distribution of species abundances, which materializes not only in term of the apparent unevenness of species abundances but also after having disentangled the genuine intensity of the underlying process driving the hierarchical structuration. All the preceding features are shared in common by the three communities studied, in line with the similarity of their environmental contexts. Only minor features can actually slightly distinguish between communities, reflecting the difference in distances between them.

Ocean warming and acidification affect the nutritional quality of the commercially-harvested turbinid snail Turbo militaris

Rising levels of atmospheric carbon dioxide are driving ocean warming and acidification. This could cause stress resulting in decreases in nutritional quality of marine species for human consumption, if environmental changes go beyond the optimal range for harvested species. To evaluate this, we used ambient and near-future elevated temperatures and pCO2 to assess impacts on the proximate nutritional composition (moisture, ash, protein, and lipids), fatty acids and trace elements of the foot tissue of Turbo militaris, a commercially harvested marine snail from south-eastern Australia. In a fully orthogonal design, the snails were exposed to ambient seawater conditions (22 ± 0.2 °C, pH 8.13 ± 0.01–450 μatm pCO2), ocean warming (25 ± 0.05 °C), pCO2 ocean acidification (pH 7.85 ± 0.02, ∼880 μatm pCO2) or a combination of both in controlled flow-through seawater mesocosms for 38 days. Moisture, ash, protein and total lipid content of the foot tissue in the turban snails was unaffected by ocean warming or acidification. However, ocean warming caused a reduction in healthful polyunsaturated fatty acids (PUFA) relative to saturated fatty acids (SFA). Under future warming and acidification conditions, there was a significant 3–5% decrease in n–3 fatty acids, which contributed to a decrease in the n–3/n–6 fatty acid ratio. The decrease in n–3 PUFAs, particularly Eicopentanoic acid (EPA), is a major negative outcome from ocean warming, because higher n–3/n–6 ratios in seafood are desirable for human health. Furthermore, ocean warming was found to increase levels of zinc in the tissues. Calcium, iron, macroelements, microelements and the composition of toxic elements did not appear to be affected by ocean climate change. Overall, the major impact from ocean climate change on seafood quality is likely to be a decrease in healthy polyunsaturated fatty acids at higher temperatures.

Differential response to Cadmium exposure by expression of a two and a three-domain metallothionein isoform in the land winkle Pomatias elegans: Valuating the marine heritage of a land snail

Through evolution, marine snails have adapted several times independently to terrestrial life. A prime example for such transitions is the adaptation to terrestrial conditions in members of the gastropod clade of Littorinoidea (Caenogastropoda). Some species of this lineage like the periwinkle (Littorina littorea), live in intertidal habitats, where they are intermittently exposed to semi-terrestrial conditions. Pomatias elegans is a close relative of Littorina littorea that has successfully colonized terrestrial habitats. Evolutionary transitions from marine to terrestrial conditions have often been fostered in marine ancestors by acquisition of physiological pre-adaptations to terrestrial life. Such pre-adaptations are based, among others, on the optimization of a wide repertoire of stress resistance mechanisms, such as the expression of metal inactivating metallothioneins (MTs).The objective of our study was to explore the Cd handling strategy in the terrestrial snail Pomatias elegans in comparison to that observed previously in Littorina littorea. After Cd exposure, the metal is accumulated mainly in the midgut gland of Pomatias elegans, in a similar way as in its marine relative. Upon Cd exposure, Pomatias elegans expresses Cd-specific MTs, as also described from Littorina littorea. In contrast to the latter species, however, the detoxification of Cd in Pomatias elegans is mediated by two different MT isoforms, one two-domain and one three-domain MT. Although the MT proteins of both species are homologous and clearly originate from one common ancestor, the three-domain MT isoform of Pomatias elegans has evolved independently from the three-domain MT of its marine counterpart, probably by addition of a third domain to the pre-existing two-domain MT. Obviously, the occurrence of homologous MT structures in both species is a hereditary character, whereas the differentiation into two distinct MT isoforms with different upregulation capacities in Pomatias elegans is an adaptive feature that probably emerged upon transition to life on land.

IDENTIFICATION OF SOME TREMATODE CERCARIAE COLLECTED FROM SIX MARINE SNAIL SPECIES OF THE GENUS NERITA.

Transmission of trematode cercariae depends mainly on the presence of marine snails as an intermediate host for zoonotic diseases. Parasitic infections in marine snails are poorly discussed. The current study was aimed to investigate the collected snails and determine the prevalence of larval trematode infections as well as identifying the cercarial types from January to December 2016. A total of 549 marine snails belonging to the genus, Nerita were randomly collected from Obhor bay, Red Sea at Jeddah city, Saudi Arabia. To harvest cercariae, snails were exposed to artificial light and the non-shedding snails were crushed. Eight distinct cercariae were identified as well as their sporocysts. Seven of them were found belonging to non-virgulate xiphidiocercaria: Ascorhytis charadriformis, cercariae, Haematoloechus similis, Litorina saxatilis V, Litorina saxatilis VII, Maritrema lingulla, Microphallus similis, Microphalliae and one ocellate furcocercous. The most prevalent type of cercariae was Maritrema lingulla (16.96%). The described cercariae are part of a diverse group of about eight trematode species inhabiting marine Nerita snails as first intermediate hosts. Such studies can assistin collecting data on disease distribution in the sympatric fish and the configuration of trematodes transmission by snails and eventually, in the avoidance and control of the subsequent fish and human diseases.

Habitual stone-tool-aided extractive foraging in white-faced capuchins, Cebus capucinus.

Habitual reliance on tool use is a marked behavioural difference between wild robust (genus Sapajus) and gracile (genus Cebus) capuchin monkeys. Despite being well studied and having a rich repertoire of social and extractive foraging traditions, Cebus sp. rarely use tools and have never been observed using stone tools. By contrast, habitual tool use by Sapajus is widespread. We review theory and discuss factors which might explain these differences in patterns of tool use between Cebus and Sapajus. We then report the first case of habitual stone tool use in a gracile capuchin: a population of white-faced capuchins (Cebus capucinus imitator) in Coiba National Park, Panama who habitually rely on hammerstone and anvil tool use to access structurally protected food items in coastal areas including Terminalia catappa seeds, hermit crabs, marine snails, terrestrial crabs and other items. This behaviour has persisted on one island in Coiba National Park since at least 2004. From 1 year of camera trapping, we found that stone tool use is strongly male-biased. Of the 205 camera trap days where tool use was recorded, adult females were never observed to use stone tools, although they were frequently recorded at the sites and engaged in scrounging behaviour. Stone tool use occurs year-round in this population; over half of all identifiable individuals were observed participating. At the most active tool use site, 83.2% of days where capuchins were sighted corresponded with tool use. Capuchins inhabiting the Coiba archipelago are highly terrestrial, under decreased predation pressure and potentially experience resource limitation compared to mainland populations—three conditions considered important for the evolution of stone tool use. White-faced capuchin tool use in Coiba National Park thus offers unique opportunities to explore the ecological drivers and evolutionary underpinnings of stone tool use in a comparative within- and between-species context.