Cephalopod behaviour.

Underlying all animal behaviors, from the simplest reflexive reactions to the more complex cognitive reasoning and social interaction, are nervous systems uniquely adapted to bodies, environments, and challenges of different animal species. Coleoid cephalopods - octopuses, squid, and cuttlefish - are widely recognized as the most behaviorally complex invertebrates and provide exciting opportunities for studying the neural control of behaviour. These unusual molluscs evolved over 400 million years ago from slow-moving armored forms to active predators of coastal and open ocean ecosystems. In this primer we will discuss how, during cephalopod evolution, the relatively simple ganglion-based molluscan nervous system has been extensively transformed to control the complex bodies and process extensive visual, tactile, and chemical sensory inputs, and summarize some recent findings about their fascinating behaviors.

Molecular characterization of cell types in the squid Loligo vulgaris.

Cephalopods are set apart from other mollusks by their advanced behavioral abilities and the complexity of their nervous systems. Because of the great evolutionary distance that separates vertebrates from cephalopods, it is evident that higher cognitive features have evolved separately in these clades despite the similarities that they share. Alongside their complex behavioral abilities, cephalopods have evolved specialized cells and tissues, such as the chromatophores for camouflage or suckers to grasp prey. Despite significant progress in genome and transcriptome sequencing, the molecular identities of cell types in cephalopods remain largely unknown. We here combine single-cell transcriptomics with in situ gene expression analysis to uncover cell type diversity in the European squid Loligo vulgaris. We describe cell types that are conserved with other phyla such as neurons, muscles, or connective tissues but also cephalopod-specific cells, such as chromatophores or sucker cells. Moreover, we investigate major components of the squid nervous system including progenitor and developing cells, differentiated cells of the brain and optic lobes, as well as sensory systems of the head. Our study provides a molecular assessment for conserved and novel cell types in cephalopods and a framework for mapping the nervous system of L. vulgaris.

Evaluation of remodeling and geometry on the biomechanical properties of nacreous bivalve shells.

Mollusks have developed a broad diversity of shelled structures to protect against challenges imposed by biological interactions(e.g., predation) and constraints (e.g., [Formula: see text]-induced ocean acidification and wave-forces). Although the study of shell biomechanical properties with nacreous microstructure has provided understanding about the role of shell integrity and functionality on mollusk performance and survival, there are no studies, to our knowledge, that delve into the variability of these properties during the mollusk ontogeny, between both shells of bivalves or across the shell length. In this study, using as a model the intertidal mussel Perumytilus purpuratus to obtain, for the first time, the mechanical properties of its shells with nacreous microstructure; we perform uniaxial compression tests oriented in three orthogonal axes corresponding to the orthotropic directions of the shell material behavior (thickness, longitudinal, and transversal). Thus, we evaluated whether the shell material's stress and strain strength and elastic modulus showed differences in mechanical behavior in mussels of different sizes, between valves, and across the shell length. Our results showed that the biomechanical properties of the material building the P. purpuratus shells are symmetrical in both valves and homogeneous across the shell length. However, uniaxial compression tests performed across the shell thickness showed that biomechanical performance depends on the shell size (aging); and that mechanical properties such as the elastic modulus, maximum stress, and strain become degraded during ontogeny. SEM observations evidenced that compression induced a tortuous fracture with a delamination effect on the aragonite mineralogical structure of the shell. Findings suggest that P. purpuratus may become vulnerable to durophagous predators and wave forces in older stages, with implications in mussel beds ecology and biodiversity of intertidal habitats.

Thermal adaptation of mRNA secondary structure: stability versus lability.

Macromolecular function commonly involves rapidly reversible alterations in three-dimensional structure (conformation). To allow these essential conformational changes, macromolecules must possess higher order structures that are appropriately balanced between rigidity and flexibility. Because of the low stabilization free energies (marginal stabilities) of macromolecule conformations, temperature changes have strong effects on conformation and, thereby, on function. As is well known for proteins, during evolution, temperature-adaptive changes in sequence foster retention of optimal marginal stability at a species' normal physiological temperatures. Here, we extend this type of analysis to messenger RNAs (mRNAs), a class of macromolecules for which the stability-lability balance has not been elucidated. We employ in silico methods to determine secondary structures and estimate changes in free energy of folding (ΔGfold) for 25 orthologous mRNAs that encode the enzyme cytosolic malate dehydrogenase in marine mollusks with adaptation temperatures spanning an almost 60 °C range. The change in free energy that occurs during formation of the ensemble of mRNA secondary structures is significantly correlated with adaptation temperature: ΔGfold values are all negative and their absolute values increase with adaptation temperature. A principal mechanism underlying these adaptations is a significant increase in synonymous guanine + cytosine substitutions with increasing temperature. These findings open up an avenue of exploration in molecular evolution and raise interesting questions about the interaction between temperature-adaptive changes in mRNA sequence and in the proteins they encode.

High biogeographic and latitudinal variability in gastropod drilling predation on molluscs along the eastern Indian coast: Implications on the history of fossil record of drillholes.

Studies on the large-scale latitudinal patterns of gastropod drilling predation reveal that predation pressure may decrease or increase with increasing latitude, or even show no trend, questioning the generality of any large-scale latitudinal or biogeographic pattern. Here, we analyze the nature of spatio-environmental and latitudinal variation in gastropod drilling along the Indian eastern coast by using 76 samples collected from 39 locations, covering ~2500 km, incorporating several ecoregions, and ~15° latitudinal extents. We find no environmental or latitudinal gradient. In fact, drilling intensity varies highly within the same latitudinal bin, or oceanic sub-basins, or even the same ecoregions. Moreover, different ecoregions with their distinctive biotic and abiotic environmental variables show similar predation intensities. However, one pattern is prevalent: some small infaunal prey taxa, living in the sandy-muddy substrate-which are preferred by the naticid gastropods-are always attacked more frequently over others, indicating taxon and size selectivity by the predators. The result suggests that the biotic and abiotic factors, known to influence drilling predation, determine only the local predation pattern. In the present case, the nature of substrate and prey composition determines the local predation intensity: soft substrate habitats host dominantly small, infaunal prey. Since the degree of spatial variability in drilling intensity within any time bin can be extremely high, sometimes greater than the variability across consecutive time bins, temporal patterns in drilling predation can never be interpreted without having detailed knowledge of the nature of this spatial variability within a time bin.

67,000 years of coastal engagement at Panga ya Saidi, eastern Africa.

The antiquity and nature of coastal resource procurement is central to understanding human evolution and adaptations to complex environments. It has become increasingly apparent in global archaeological studies that the timing, characteristics, and trajectories of coastal resource use are highly variable. Within Africa, discussions of these issues have largely been based on the archaeological record from the south and northeast of the continent, with little evidence from eastern coastal areas leaving significant spatial and temporal gaps in our knowledge. Here, we present data from Panga ya Saidi, a limestone cave complex located 15 km from the modern Kenyan coast, which represents the first long-term sequence of coastal engagement from eastern Africa. Rather than attempting to distinguish between coastal resource use and coastal adaptations, we focus on coastal engagement as a means of characterising human relationships with marine environments and resources from this inland location. We use aquatic mollusc data spanning the past 67,000 years to document shifts in the acquisition, transportation, and discard of these materials, to better understand long-term trends in coastal engagement. Our results show pulses of coastal engagement beginning with low-intensity symbolism, and culminating in the consistent low-level transport of marine and freshwater food resources, emphasising a diverse relationship through time. Panga ya Saidi has the oldest stratified evidence of marine engagement in eastern Africa, and is the only site in Africa which documents coastal resources from the Late Pleistocene through the Holocene, highlighting the potential archaeological importance of peri-coastal sites to debates about marine resource relationships.

Application of biological and fisheries attributes to assess the vulnerability and resilience of tropical marine fish species.

Taking advantage of published data on life-history traits and short-term information on fishery parameters from 3132 records for 644 fish stocks along the coast of India, we calculated resilience (R) and vulnerability (V). Further, we developed an Index of Resilience and Vulnerability (IRV) for 133 species of tropical finfishes, crustaceans, and molluscs. Using 7 resilience and 6 vulnerability attributes, two-dimensional scatter plots of the resilience and vulnerability scores were generated and the Euclidean distance and angle from the origin to each point were calculated to determine IRV and the effect of fishing on fish species. By ranking the species, the top 10 highly resilient, highly vulnerable, and high-risk species (low IRV) were identified. While small-sized species with fast growth rate and low trophic level were among the highly resilient species, large predatory species such as sharks and barracudas were among the highly vulnerable and high-risk species. More than 100 of the 133 species were resilient-yet-vulnerable, and most crustaceans showed high resilience. Differences in IRV scores among species within the same family were discernible, indicating the differences in the biological characteristics and response to fishing. Sensitivity analysis indicated that an abridged IRV with 6 attributes works similar to 13 attributes and can be used in data-deficient situations. Comparison of R and V of IRV with other assessments showed different results because of divergences in the objectives, number and types of attributes, and thresholds used. These assessments do not convey the same information and therefore great care must be taken for reproducing these frameworks to other fisheries. The results of IRV analysis can be useful for stock assessments and in developing effective management measures in combination with other complementary information.

Specialized predation by deep-sea Solenogastres revealed by sequencing of gut contents.

'Who eats whom?' is a fundamental question about the ecological roles and interactions of deep-sea organisms. However, the tools needed to analyze trophic relationships remain limited, especially with regard to studying meiofauna and small macrofauna in abyssal and hadal depths. We present results from indirect molecular analyses of the gut contents of abyssal and hadal Solenogastres (Mollusca, Aplacophora) of the Northwest Pacific. Our data revealed a high food specialization and a surprising diversity of food sources among these inconspicuous worm-shaped predators. We hypothesize that Hydrozoa forms the ancestral food source of Solenogastres, and that specialization on non-cnidarian prey (such as annelids, nemerteans, and bivalves) evolved independently along with modifications in the digestive tract. Despite being intuitively advantageous in the nutrient-limited deep sea, we found only one widespread generalist feeder (potentially associated with scavenging).

Non-collinear Hox gene expression in bivalves and the evolution of morphological novelties in mollusks.

Hox genes are key developmental regulators that are involved in establishing morphological features during animal ontogeny. They are commonly expressed along the anterior-posterior axis in a staggered, or collinear, fashion. In mollusks, the repertoire of body plans is widely diverse and current data suggest their involvement during development of landmark morphological traits in Conchifera, one of the two major lineages that comprises those taxa that originated from a uni-shelled ancestor (Monoplacophora, Gastropoda, Cephalopoda, Scaphopoda, Bivalvia). For most clades, and bivalves in particular, data on Hox gene expression throughout ontogeny are scarce. We thus investigated Hox expression during development of the quagga mussel, Dreissena rostriformis, to elucidate to which degree they might contribute to specific phenotypic traits as in other conchiferans. The Hox/ParaHox complement of Mollusca typically comprises 14 genes, 13 of which are present in bivalve genomes including Dreissena. We describe here expression of 9 Hox genes and the ParaHox gene Xlox during Dreissena development. Hox expression in Dreissena is first detected in the gastrula stage with widely overlapping expression domains of most genes. In the trochophore stage, Hox gene expression shifts towards more compact, largely mesodermal domains. Only few of these domains can be assigned to specific developing morphological structures such as Hox1 in the shell field and Xlox in the hindgut. We did not find traces of spatial or temporal staggered expression of Hox genes in Dreissena. Our data support the notion that Hox gene expression has been coopted independently, and to varying degrees, into lineage-specific structures in the respective conchiferan clades. The non-collinear mode of Hox expression in Dreissena might be a result of the low degree of body plan regionalization along the bivalve anterior-posterior axis as exemplified by the lack of key morphological traits such as a distinct head, cephalic tentacles, radula apparatus, and a simplified central nervous system.

Time dependent differential regulation of a novel long non-coding natural antisense RNA during long-term memory formation.

Long natural antisense transcripts (NATs) have been demonstrated in significant numbers in a variety of eukaryotic organisms. They are particularly prevalent in the nervous system suggesting their importance in neural functions. However, the precise physiological roles of the overwhelming majority of long NATs remain unclear. Here we report on the characterization of a novel molluscan nitric oxide synthase (NOS)-related long non-coding NAT (Lym-NOS1AS). This NAT is spliced and polyadenylated and is transcribed from the non-template strand of the Lym-NOS1 gene. We demonstrate that the Lym-NOS1AS is co-expressed with the sense Lym-NOS1 mRNA in a key neuron of memory network. Also, we report that the Lym-NOS1AS is temporally and spatially regulated by one-trial conditioning leading to long term memory (LTM) formation. Specifically, in the cerebral, but not in the buccal ganglia, the temporal pattern of changes in Lym-NOS1AS expression after training correlates with the alteration of memory lapse and non-lapse periods. Our data suggest that the Lym-NOS1AS plays a role in the consolidation of nitric oxide-dependent LTM.

Use of a gene encoding zona pellucida 4 as a female-specific marker for early stage sexual differentiation and size dimorphism in the pacific abalone Haliotis discus hannai.

Growth rates of Pacific abalone Haliotis discus hannai are an important trait affecting the economic value in the abalone aquaculture industry. A reverse-transcription polymerase chain reaction (RT-PCR) analyses of tissues from H. discus hannai was conducted for sexually mature gonads to determine male- and female-specific target gene expression, including genes encoding zona pellucida domain 4 (zp4), sperm protein (sp) and lysin (lys), respectively. Sex-specific expression patterns of these gene expression, even in sexually immature abalone, indicate these genes can be used as sensitive and robust sex-specific molecular markers. The RT-PCR procedure was also performed to analyze tissues collected at various developmental stages (50-day intervals) beginning at fertilization to determine when sex differentiation and expression of sex-specific genes was initiated. Detection of zp4 transcript in tissues collected at 200 days post-fertilization (dpf) indicated egg-specific development starts at 150-200 dpf. To evaluate possible sex-specific differences in growth rate, there was conducting of a molecular marker-based sex identification of abalone from a population selected for rapid growth rate. In a group of large H. discus hannai, females were more prevalent than males. To assess the correlation between growth and sex, there was comparison of weights of 3-year-old Pacific abalone in specimens where there had been sex determinations by visual examination and molecular methods. The results indicated females weighed more (55.92 ± 9.38 g, n = 15) than males (43.64 ± 15.55 g, n = 6, P = 0.037), indicating females had a more rapid growth rate than males.

Advancing d-amino acid-containing peptide discovery in the metazoan.

The discovery of enzyme-derived d-amino acid-containing peptides (DAACPs) that have physiological importance in the metazoan challenges previous assumptions about the homochirality of animal proteins while simultaneously revealing new analytical challenges in the structural and functional characterization of peptides. Most known DAACPs have been identified though laborious activity-guided purification studies or by homology to previously identified DAACPs. Peptide characterization experiments are increasingly dominated by high throughput mass spectrometry-based peptidomics, with stereochemistry rarely considered due to the technical challenges of identifying l/d isomerization. This review discusses the prevalence of enzyme-derived DAACPs among animals and the physiological consequences of peptide isomerization. Also highlighted are the analytical methods that have been applied for structural characterization/discovery of DAACPs, including results of several recent studies using non-targeted discovery methods for revealing novel DAACPs, strongly suggesting that more DAACPs remain to be uncovered.

A Review of Pedal Peptide/Orcokinin-type Neuropeptides.

Neuropeptides are endogenous active substances that play important roles in a number of physiological processes and are ubiquitous in the nervous tissue in vivo. The gene encoding pedal peptide/orcokinin-type (PP/OK-type) neuropeptide is an important member of the neuropeptide gene family and is ubiquitous in invertebrates of Bilateria; orcokinin (OK) is mainly found in Arthropoda, while pedal peptide (PP) is mainly found in Mollusca. OK and PP are also present in other animals. PP/OK-type neuropeptides are a kind of multifunctional neuropeptides predominantly expressed in the nervous tissue and play important roles in the nerve regulation of movement. Moreover, OK has a number of other physiological functions. This review describes the distribution, expression, function and maturation of PP/OK-type neuropeptides to facilitate investigations of new functions and receptors of PP/OK-type neuropeptides, providing the theoretical foundation for the potential use of PP/OK-type neuropeptides in the prevention and control of agricultural and forestry pests, as an additive for skin care products and in the screening of drugs for the treatment of diabetes.

Mechanism and Function of the Catch State in Molluscan Smooth Muscle: A Historical Perspective.

Molluscan smooth muscles exhibit the catch state, in which both tension and resistance to stretch are maintained with very low rates of energy consumption. The catch state is studied mainly on the anterior byssus retractor muscle (ABRM) of a bivalve molluscan animal, Mytilus, which can easily be split into small bundles consisting of parallel fibers. The ABRM contracts actively with an increase in the intracellular free Ca ion concentration, [Ca2+]i, as with all other types of muscle. Meanwhile, the catch state is established after the reduction of [Ca2+]i to the resting level. Despite extensive studies, the mechanism underlying the catch state is not yet fully understood. This article briefly deals with (1) anatomical and ultrastructural aspects of the ABRM, (2) mechanical studies on the transition from the active to the catch state in the isotonic condition, (3) electron microscopic and histochemical studies on the intracellular translocation of Ca ions during the transition from the active to the catch state, and (4) biochemical studies on the catch state, with special reference to a high molecular mass protein, twitchin, which is known to occur in molluscan catch muscles.

The Pleistocene-Holocene aquatic molluscs as indicators of the past ecosystem changes in Transbaikalia (Eastern Siberia, Russia).

Data on the historical change of the Transbaikalian malacofauna in the Neopleistocene and Holocene is presented. Aquatic mollusc shells from archaeological excavations of the ancient settlements dating from the Neolithic period to Medieval and also from a drill hole of the Neopleistocene alluvial deposits were collected. In total eight species of bivalve molluscs from the families Margaritiferidae, Unionidae, Lymnocardiidae, Glycymerididae [marine], and two gastropod species from families Viviparidae and Planorbidae were identified. These species were aged using radiocarbon dating. It was found that the species ranged in age from more than 50.000 to 2.080-1.210 years BP. Five species inhabited the Transbaikal region which are locally extirpated today. Their disjunctive ranges in the past included southern Europe and Western and Eastern Siberia to Transbaikalia and in the east to Far East and Primorye Territory of Russia. A remarkable finding is that of the bivalve genus Monodacna, which was found very far from its native range, the Ponto-Caspian region. The time of existence and extirpation of the thermophilic species of genera Monodacna, Planorbis, Lanceolaria and Amuropaludina corresponds to cycles of the warming and cooling in Pleistocene and Holocene according to regional climate chronological scales. These species can be used as palaeoclimate indicators. Change of the regional malacofaunal species composition is connected with the natural climatochron cycles in the Pleistocene and Holocene resulting in evidence for succession. In the course of this succession, these stenothermal species became extirpated on a regional level, decreasing their global ranges.

Effects of UV-B radiation on secondary metabolite production, antioxidant activity, photosynthesis and herbivory interactions in Nymphoides humboldtiana (Menyanthaceae).

Ultraviolet B-light (UV-B) can exert indirect effects on plant-herbivore interactions by inducing changes in constitutive and induced chemical defenses, since it modulates physiological aspects of plants. This study evaluated the action of UV-B radiation on photosynthesis and production of secondary metabolites in Nymphoides humboldtiana and the cascade effects on the relationship of this macrophyte with a generalist herbivore, the gastropod mollusk Biomphalaria glabrata. After 13 days of UV-B exposition under laboratory conditions, the floating macrophyte N. humboldtiana responded increasing its photosynthetic potential and the production of flavonoids with a correlated enhance in antioxidant activity. However, these changes observed in its metabolism were not enough to alter their palatability to consumption by B. glabrata verified through laboratory feeding choice experiments. Despite the known deleterious effects of exposure to UV-B on terrestrial plants, we found that N. humboldtiana does have physiological/biochemical mechanisms as a strategy or restorative response to this potencially adverse or impacting agent without changing its relationships with herbivores.

Not just scratching the surface: distinct radular motion patterns in Mollusca.

The radula is the organ for mechanical food processing and an important autapomorphy of Mollusca. Its chitinous membrane, embedding small radular teeth, is moved by the set of muscles resulting in an interaction with the ingesta, tearing it and collecting loosened particles. Radulae and their teeth can be quite distinct in their morphology and had been of high research interest, but only a few studies have examined the basic functional principles of this organ, the movement and motion during feeding action. Here, the radular motion of 20 representative species, belonging to four major gastropod lineages (Vetigastropoda, Neritimorpha, Caenogastropoda and Heterobranchia) and Polyplacophora, were recorded and classified. Comparisons of the video footage with the scanning electron microscope (SEM) images of the radula resulted in the recognition of functional tooth rows and the correct position of the teeth during feeding. We identified six different types of radular movements, including rotations and bending of the radula itself. In each movement type, different structures act as counter bearings enabling the animals to grab and tear food.

Genes with spiralian-specific protein motifs are expressed in spiralian ciliary bands.

Spiralia is a large, ancient and diverse clade of animals, with a conserved early developmental program but diverse larval and adult morphologies. One trait shared by many spiralians is the presence of ciliary bands used for locomotion and feeding. To learn more about spiralian-specific traits we have examined the expression of 20 genes with protein motifs that are strongly conserved within the Spiralia, but not detectable outside of it. Here, we show that two of these are specifically expressed in the main ciliary band of the mollusc Tritia (also known as Ilyanassa). Their expression patterns in representative species from five more spiralian phyla-the annelids, nemerteans, phoronids, brachiopods and rotifers-show that at least one of these, lophotrochin, has a conserved and specific role in particular ciliated structures, most consistently in ciliary bands. These results highlight the potential importance of lineage-specific genes or protein motifs for understanding traits shared across ancient lineages.

Biological responses to extreme weather events are detectable but difficult to formally attribute to anthropogenic climate change.

As the frequency and intensity of extreme events such as droughts, heatwaves and floods have increased over recent decades, more extreme biological responses are being reported, and there is widespread interest in attributing such responses to anthropogenic climate change. However, the formal detection and attribution of biological responses to climate change is associated with many challenges. We illustrate these challenges with data from the Elbe River floodplain, Germany. Using community turnover and stability indices, we show that responses in plant, carabid and mollusc communities are detectable following extreme events. Community composition and species dominance changed following the extreme flood and summer heatwave of 2002/2003 (all taxa); the 2006 flood and heatwave (molluscs); and after the recurring floods and heatwave of 2010 and the 2013 flood (plants). Nevertheless, our ability to attribute these responses to anthropogenic climate change is limited by high natural variability in climate and biological data; lack of long-term data and replication, and the effects of multiple events. Without better understanding of the mechanisms behind change and the interactions, feedbacks and potentially lagged responses, multiple-driver attribution is unlikely. We discuss whether formal detection and/or attribution is necessary and suggest ways in which understanding of biological responses to extreme events could progress.

Geriatric Invertebrates.

Invertebrates are becoming more popular and, as collections age, clients may seek veterinary intervention where the welfare of the animal must be considered. This article covers aging in many invertebrate species but with a focus on species likely to be seen in general practice. Supportive care may be an option to prolong life, but euthanasia must be considered for invertebrates with age-related unmanageable conditions.