Microgeographical variation in shell strength in the flat periwinkles Littorina obtusata and Littorina mariae

The strength of molluscan shells has been shown to vary in adaptive ways in a number of species and one of the main factors thought to be involved is shell-crushing by predators. A recent study found that the sibling species of flat periwinkle Littorina obtusata and Littorina mariae showed significant differences in the rates at which shell strength increased with shell length in specimens which had been collected from the same location, where the species were sympatric. This paper describes differences between the shells of the two species from a number of localities around Milford Haven in Dyfed, Wales, and local geographical variation in the shells. Littorina mariae, which is normally found at lower tidal levels than L. obtusata, matures at a smaller shell length. Both species reinforce the shell as they grow since shell strength, determined as the maximum force applied by a hydraulic tensile testing machine before the shell cracked, is strongly positively allometric; it increases at a rate close to the cube of shell length whilst isometric growth would result in strength increasing in proportion to the square of shell length. Because L. mariae matures earlier and reinforces the shell at a smaller size, the mature shell of L. mariae is substantially stronger on average than that of a similar sized but immature L. obtusata. At maturity the shell strengths of the two species are not very different despite the substantial difference in mean shell length. Strength varies significantly from shore to shore, and with the level of the shore from which the animals were collected. Strength increases down the shore in both species. Shell strength decreases with exposure to wave action in L. mariae but increases with exposure in L. obtusata; there is also substantial shore-to-shore variation which is not explained by exposure. Path analysis was used to explore the relationship between shell strength and other measured shell parameters (mass, length, height, thickness). The best predictor of shell strength in both species is a parameter which is heavily positively loaded on LN (shell mass) and strongly offset by negative loadings on LN (shell length) and LN (shell height). This is logical because for a given shell length a heavier shell will be thicker and stronger, whilst for a given shell mass a bigger shell will be thinner and therefore weaker. Such differential variation of shell mass and shell length explains most of the geographical variation observed in shell strength; shells are stronger in snails collected from one place than from another because, for the same shell length they are heavier or, to put it the other way, because at the same shell mass, they are smaller.     

The relationship between flat periwinkle life histories and digenean infections

Larval digenean parasites were studied in Littorina obtusata (L.) and L. mariae Sacchi & Rastelli at Sawdern Point in West Wales. Shell parameters, ovipositor and shell colour, penis morphology and sex ratios were scored, and the influence of parasitism studied. A total of 7 species of parasites were found, although the prevalence was very low in L. mariae, especially in the females. The parasitic gigantism that has been described in other species of gastropod was not found in this study. Parasitic castration does occur in some infected male L. obtusata, resulting in severe stunting of the penis. However, this phenomena was never observed in L. mariae. The winkle species are congeneric and inhabit broadly similar niches on the shore; their life histories are however quite different — one being annual and the other perennial. This probably affects exposure to infection, and might explain the differing prevalence of the parasites.     

Complete lack of mitochondrial divergence between two species of NE Atlantic marine intertidal gastropods

Some mitochondrial introgression is common between closely related species, but distinct species rarely show substantial introgression in their entire distribution range. In this study, however, we report a complete lack of mitochondrial divergence between two sympatric species of flat periwinkles (Littorina fabalis and Littorina obtusata) which, based on previous allozyme studies, diverged approximately 1 Ma. We re‐examined their species status using both morphology (morphometric analysis) and neutral genetic markers (microsatellites) and our results confirmed that these species are well separated. Despite this, the two species shared all common cytochrome‐b haplotypes throughout their NE Atlantic distribution and no deep split between typical L. fabalis and L. obtusata haplotypes could be found. We suggest that incomplete lineage sorting explains most of the lack of mitochondrial divergence between these species. However, coalescent‐based analyses and the sympatric sharing of unique haplotypes suggest that introgressive hybridization also has occurred.     

Maintenance of zonation patterns in two species of flat periwinkle,Littorina obtusata and L. mariae

The zonation patterns of Littorina obtusata (L.) and Littorina mariae Sacchi et Rastelli were shown to be quite distinct on a sheltered rocky shore. L. obtusata was found at all the heights sampled; it reached peak numbers at mid shore on the alga Ascophyllum nodosum L. (Le Jol). There was no difference in the tidal height occupied by adults or juveniles; or in the mean size of L. obtusata along the vertical gradient of the shore. In contrast L. mariae occurred exclusively low on the shore, on Fucus serratus L. Translocation of the two species within their respective levels resulted in random movement after 4 days, although initial movements after 1 and 2 days were sometimes directional. Animals transplanted to the normal level of the other species showed directional movement towards their home zone; this was most pronounced after 4 days. There was no difference in the distance moved by the two species, although the distance moved did vary with tidal height, both species moving further at mid shore than low shore. Distances moved by littorinids at replicate areas in the low shore were similar but those at mid shore did vary. There was an interaction between the species and the different tidal heights which revealed that transplanted species moved further than translocated species at the same tidal level. However, this was only significant in the case of L. mariae. It is suggested that the close relationship between the winkles and their host algae may direct the homing behaviour of displaced individuals.     

Lack of metabolic temperature compensation in the intertidal gastropods,Littorina saxatilis (Olivi) and L. obtusata (L.)

Two intertidal snails, Littorina saxatilis (Olivi, 1972) (upper eulittoral fringe/maritime zone) and Littorina obtusata (Linnaeus, 1758) (lower eulittoral) were collected from a boulder shore on Nobska Point, Cape Cod, Massachusetts, in July and acclimated for 15–20 days at 4 ° or 21 °C. Oxygen consumption rate (Vo₂) was determined for 11–15 subsamples of individuals at 4 °, 11 ° and 21 °C with silver/platinum oxygen electrodes. Multiple factor analysis of variance (MFANOVA) of lo₁₀ transformed values of whole animal Vo₂ with log₁₀ dry tissue weight (DTW) as a covariant revealed that increased test temperature induced a significant increase in Vo₂ in both species (P<0.00001). In contrast, MFANOVA revealed that temperature acclimation did not affect Vo₂ in either L. saxatilis (P= 0.35) or L. obtusata (P= 0.095). Thus, neither species displayed a capacity for the typical metabolic temperature compensation marked by an increase in Vo₂ at any one test temperature in individuals acclimated to a lower temperature that is characteristic of most ectothermic animals. Lack of capacity for metabolic temperature acclimation has also been reported in other littorinid snail species, and may be characteristic of the group as a whole. Lack of capacity for respiratory temperature acclimation in these two species and other littorinids may reflect the extensive semi-diurnal temperature variation that they are exposed to in their eulittoral and eulittoral fringe/maritime zone habitats. In these habitats, any metabolic benefits derived from longer-term temperature compensation of metabolic rates are negated by extreme daily temperature fluctuations. Instead, littorinid species appear to have evolved mechanisms for immediate metabolic regulation which, in L. saxatilis and L. obtusata and other littorinids, appear to centre on a unique ability for near instantaneous suppression of metabolic rate and entrance into short-term metabolic diapause at temperatures above 20–35 °C, making typical seasonal respiratory compensation mechanisms characteristic of most ectotherms of little adaptive value to littorinid species.     

Resistance to freshwater exposure in White Sea Littorina spp. I: Anaerobic metabolism and energetics

 Anaerobic metabolism and changes in the osmotic concentration of extravisceral fluid were studied in the White Sea periwinkles (Littorina littorea, Littorina saxatilis and Littorina obtusata) during freshwater exposure. Resistance to hypoosmotic stress increased in the order: L. obtusata < L. saxatilis < L. littorea. Our data suggest that osmotic shock is not a primary reason for mortality of the periwinkles under these conditions. During environmental anaerobiosis, considerable succinate accumulation (up to 10–19 μmol g⁻¹ wet weight), and depletion of phosphagen and ATP pools were found in the studied species. Other metabolic end products (alanopine, strombine, lactate, acetate or propionate) were not detected. Succinate accumulation and net ATP breakdown were the fastest in the least resistant species, L. obtusata, and slowest in the most resistant, L. littorea. Rate of ATP turnover decreased during freshwater exposure in L. littorea and L. saxatilis, but not in L. obtusata. Our data suggest that differential resistance of three studied Littorina spp. to extreme hypoosmotic stress may be related to their different abilities to reduce metabolic rate and ATP turnover during sustained anoxia. Species-specific variations in anaerobic capacity of Littorina spp. are discussed in relation to their vertical distribution, size and ecology. Accepted: 4 October 1999     

Selective predation favouring cryptic individuals of marine snails (Littorina)

Many gastropods have inherited conspicuous shell colour polymorphisms. A challenging question is, are colour frequencies under selection or is polymorphism owing to random evolutionary processes? The intertidal species Littorina saxatilis (a rock‐dweller) and L. obtusata (confined to macroalgae) both have genetically determined shell colour variation. In Iceland, Littorina obtusata are mostly cryptic on brown macroalgae by having brown or yellow shells (~95% of the snails), while Littorina saxatilis often appears conspicuous to the background of dark rocks owing to non‐cryptic colours (15–20%). This difference may be due to selective elimination of conspicuously coloured L. obtusata by visual predators, while L. saxatilis, largely living in another habitat, is not under a similarly intense colour selection. To test this hypothesis we increased the frequencies of conspicuous L. saxatilis in experimental populations (from <12 to 55%) and placed these in the seaweed zone, the main habitat of L. obtusata. Fifteen populations were released on isolated spots of seaweed and three of these were covered by net cages to exclude bird predators. One month later, yellow snails had increased in frequency within the patches, and to our surprise the result did not differ between bare and caged patches. This suggests selection favouring a colour that matches the background of fucoid seaweeds by visual predators able to enter the cages. Birds acted as important predators by picking 16% of the experimental snails in the uncaged spots, but were unable to enter the caged spots. However, the bird predation was non‐selective with respect to snail colour. For various reasons the most likely predators able to enter the cages were intertidal fish, these were thus responsible for the selection of non‐cryptic snails. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society, 2002, 76 , 137–144.     

The habitat and feeding preferences of Littorina obtusata (L.) and L. mariae sacchi et rastelli

The vertical zonation of the flat periwinkles Littorina obtusata (L.) and L. mariae Sacchi et Rastelli is different, but overlaps. At a sheltered location on the Isle of Cumbrae, Scotland, L. obtusata, although most abundant on Fucus vesiculosus L. and Ascophyllum nodosum (L.) Le Jol. on the midshore, ranged over the entire shore from the Pelvetia zone down to the upper limits of Laminaria. Littorina mariae exhibited a much more restricted distribution being associated predominantly with Fucus serratus L. on the lower shore. In laboratory trials with adult plants, flat periwinkles found fucoid algae both more attractive and more palatable than other seaweeds. Littorina obtusata also preferred the reproductive receptacles of fucoid algae to their vegetative tissue. Tiny germlings of a diversity of seaweeds were readily consumed by L. obtusata. In choice experiments germling Ulva lactuca L. was preferred to adult U. lactuca, whereas adult Fucus serratus was preferred to germling F. serratus. Contrasting foraging behaviours between different species of flat periwinkle are discussed in relation to differences in their radular dentition.     

Characterization of polymorphic microsatellites for the periwinkle gastropod, Littorina littorea (Linnaeus, 1758) and their cross-amplification in four congeners

Eight polymorphic microsatellite loci are described for Littorina littorea (Linnaeus, 1758). Data on allelic variation in Irish and Celtic Sea samples are reported. The average number of alleles per locus was 11 (range 4–29), and observed and expected heterozygosities ranged from 6.9 to 84.3% and from 9.4 to 95.2%, respectively. Loci did not deviate from Hardy–Weinberg equilibrium and no linkage disequilibrium between loci pairs was detected. Microsatellites were not highly conserved in the congeners, L. fabalis, L. saxatilis, L. compressa and L. obtusata as evidenced by a low rate of cross-amplification. These microsatellites should prove useful in population genetic studies.     

Method of color measurements of the elements of color in gastropods shell

The method of the accurate determination of the shell color of gastropods has been studied with the example of the gastropod Littorina obtusata. The suggested method is based on using the system of MKO RGB color coordinates and makes it possible to conduct measurements of both color as a whole and the color of the individual elements of the shell picture.     

Crystalline calcium in littorinid mucus trails

Previous work has shown that the feet of terrestrial and freshwater snails are important in calcium regulation, often secreting granules of CaCO₃. This phenomenon has not, until now, been observed in marine snails. Here we report the presence of CaCO₃ granules in the trail mucus of Littorina littorea (L.), L. saxatilis (Olivi) and L. obtusata (L.) Fixed mucus trails on plastic coverslips were examined by X-ray microanalysis under the SEM. Of the single-metal granules observed in the mucus trails the most abundant were of calcium (means: L. littorea, 440 mm⁻²; L. saxatilis, 401 mm⁻²; L. obtusata, 348 mm⁻²) followed for each species by silicon (maximum mean density: L. saxatilis, 120 mm⁻²) and iron (maximum mean density: L. saxatilis, 65 mm⁻²) granules. Single-metal granules of Al, Ti, Mg and P were also found but only in the mucus trails of L. obtusata, perhaps reflecting its different collection site from the other two species. The mean size of the calcium granules showed significant interspecific variation (L. littorea, 1.32 μm diameter±0 08 μm, n = 143; L. saxatilis, 1.80 μm±0.12, n = 113; L. obtusata, 2.14 μm±0.09, n = 167). Most calcium granules (L. littorea, 80%, n = 35; L. saxatilis, 57%, n = 113; L. obtusata, 69%, n = 167) were attached to, or embedded within, microthreads of mucus which tended to run parallel to the direction of locomotion. The significance of this is unknown although it may imply that the CaCO₃ granules are secreted with the mucus. It is concluded that calcium losses via this route are too small for pedal mucus to function significantly in ionoregulation of calcium. The calcium in the trail may therefore perform other functions, for example indicating trail polarity.     

THE EGG MASS OF LITTORINA OBTUSATA AND LACUNA PALLIDULA (GASTROPODA : PROSOBRANCHIA)

The egg masses of Littorina obtusata and of Lacuna pallidulahave been compared with regard to size, shape, egg number, eggsize and substratum. The structure and histochemistry of thespawn of the two species has been investigated and observationson the laying process in L. obtusata are also recorded. ^* Present address: Portway School, Penpole Lane, Shirehampion,Bristol, BS11 0EB. (Received 12 October 1977;     

Effects of ambient temperature and of temperature acclimation on nitrogen excretion and differential catabolism of protein and nonprotein resources in the intertidal snails,Littorina saxatilis (Olivi) and L. obtusata (L.)

Nitrogenous excretion in two snails, Littorina saxatilis (high intertidal) and L. obtusata (low intertidal) was studied in relation to temperature acclimation (at 4° and 21°C), including total N excretion rates, the fraction of urea in N excretion, corresponding O:N ratios and the partitioning of deaminated protein between catabolic and anabolic processes at 4°, 11° and 21°C. Aggregate N excretion rates in both species showed no significant compensatory adjustments following acclimation. Total weight specific N excretion rates at 21°C were higher in standard 3 mg L. saxatilis (739 ng N mg⁻¹ h⁻¹) than standard 5 mg L. obtusata (257 ng N mg⁻¹ h⁻¹) for snails acclimated to 21°C. Comparisons of Q₁₀ values of total weight specific N excretion to Q₁₀ values for weight specific oxygen consumption ({xxV}O₂) between 4° to 11 °C and 11° to 21°C indicated that, while total rates of catabolic metabolism ({xxV}O₂) and protein deamination in L. obtusata were essentially parallel, the relationship between N excretion and {xxV}O₂ in L. saxatilis revealed the partitioning of a larger share of deaminated protein carbon into anabolism at 4° and 21°C than at 11°C. Urea N accounted for a larger share of aggregate N excreted in L. saxatilis than in L. obtusata, but in both species urea N is a greater proportion of total N excreted when acclimated at 4°C (urea N: ammonia N ratio range: 1 to 2.15) than in snails acclimated to 21°C (urea N: ammonia N ratio range: 0.46 to 1.39). Molar O:N ratios indicate that the proportion of metabolism supported by protein catabolism is greater in L. saxatilis (O:N range: 2.5–8.4) than in L. obtusata (O:N range: 7.3–13.0). In both species, regardless of acclimation temperature, the O:N ratios are generally lowest (high protein catabolism) at 4°C and highest at 21°C.     

Inheritance of longitudinal shell bands in the snails <Emphasis Type="Italic">Littorina obtusata</Emphasis> and <Emphasis Type="Italic">L. saxatilis</Emphasis> (Gastropoda,Prosobranchia)

The hypothesis of a monogenic inheritance of dark longitudinal bands on the shell in the gastropods Littorina obtusata and L. saxatilis was checked. One gene having two alleles proved to be responsible for the shell banding pattern in both of the species. The presence of bands was a dominant character in either case.     

Temporal dynamics of inducible anti‐herbivory defenses in the brown seaweed Ascophyllum nodosum (Phaeophyceae)1

Anti‐herbivory defenses support persistence of seaweeds. Little is known, however, about temporal dynamics in the induction of grazer‐deterrent seaweed traits. In two induction experiments, consumption rates of the periwinkle Littorina obtusata (L.) on the brown seaweed Ascophyllum nodosum (L.) Le Jolis were measured in 3‐d intervals. Changes in palatability of directly grazed A. nodosum were tested every 3 d with feeding assays using fresh and reconstituted seaweed pieces. Likewise, assays with fresh A. nodosum assessed changes in seaweed palatability in response to water‐borne cues from nearby grazed conspecifics. Consumption rates of L. obtusata varied significantly during the 27‐d induction phase of each experiment. Direct grazing by L. obtusata lowered palatability of fresh and reconstituted A. nodosum pieces to conspecific grazers after 15 d as well as after 6 and 12 d, respectively. After 12, 18, and 24 d, fresh A. nodosum located downstream of L. obtusata‐grazed conspecifics was significantly less palatable than A. nodosum located downstream of ungrazed conspecifics. Changes in L. obtusata consumption rates and A. nodosum palatability during both induction experiments suggest temporal variation of grazer‐deterrent responses, which may complicate experimental detection of inducible anti‐herbivory defenses.     

ON THE OCCURRENCE OF THE PROSOBRANCHS LITTORINA NERITOIDES, L. MARIAE AND L. OBTUSATA IN DENMARK

Littorina neritoides occurs on piers and large stones on thebeaches in northern Jutland, but may not be able to toleratecold winters. L. obtusata and L. mariae are common on Fucus,but only L. mariae penetrates deeply into the fjords and sounds. (Received 10 December 1979;     

Predatory behaviour of juvenile shore crab Carcinus maenas (L.)

Juvenile shore crabs Carcinus maenas (L.) were observed feeding on rock barnacles Semibalanus balanoides (L.) on a Bay of Fundy rocky shore. This previously unreported predatory behaviour was further investigated in the laboratory. When given a choice of three common and abundant gastropods, Nucella lapillus (L.), Littorina littorea (L.), and Littorina obtusata (L.), and the rock barnacle Semibalanus balanoides, juvenile shore crabs of both sexes ate mainly barnacles and consumed proportionately more barnacles than gastropods compared with adults, which ate mainly gastropods. The rock barnacle is an abundant and readily available food source which may be important in sustaining the juvenile crab through periods of moults and rapid growth. As the shore crab attains a certain age (size), it must forage lower on the shore as gastropods become more important in its diet.     

Distribution of the species of rough periwinkle (Littorina) in Great Britain

The status of the four currently recognised species in the Littorina saxatilis species-complex, i.e. Littorina nigrolineata Gray, L. arcana Hannaford Ellis, L. saxatilis (Olivi) and L. neglecta Bean is reviewed briefly, with notes on their characteristic features and location on the shore. Since the taxonomy of these rough periwinkles has only become stable relatively recently much of the previously published information on distribution is of little or no use. In this paper their distribution around the coastline of England and Wales (with some notes on Scotland) is described and discussed. L. saxatilis is found in a wide range of habitats from exposed peninsulas to estuaries on all shores where there is a suitable rocky or stony substrate; also in salt marsh pools. L. arcana has a more restricted distribution and is notably absent along much of the south English coast, central Cardigan Bay and possibly northern Scotland; it is not found in estuaries. L. nigrolineata has an even more restricted distribution, although it occurs both on exposed coasts and in estuaries; it has only been found by us in one locality on the east coast. L. neglecta is probably fairly widely distributed but we have few details so far. The implications of the different patterns of distribution are discussed.     

Hybridization patterns between two marine snails,Littorina fabalis and L. obtusata

Characterizing the patterns of hybridization between closely related species is crucial to understand the role of gene flow in speciation. In particular, systems comprising multiple contacts between sister species offer an outstanding opportunity to investigate how reproductive isolation varies with environmental conditions, demography and geographic contexts of divergence. The flat periwinkles, Littorina obtusata and L. fabalis (Gastropoda), are two intertidal sister species with marked ecological differences compatible with late stages of speciation. Although hybridization between the two was previously suggested, its extent across the Atlantic shores of Europe remained largely unknown. Here, we combined genetic (microsatellites and mtDNA) and morphological data (shell and male genital morphology) from multiple populations of flat periwinkles in north‐western Iberia to assess the extent of current and past hybridization between L. obtusata and L. fabalis under two contrasting geographic settings of divergence (sympatry and allopatry). Hybridization signatures based on both mtDNA and microsatellites were stronger in sympatric sites, although evidence for recent extensive admixture was found in a single location. Misidentification of individuals into species based on shell morphology was higher in sympatric than in allopatric sites. However, despite hybridization, species distinctiveness based on this phenotypic trait together with male genital morphology remained relatively high. The observed variation in the extent of hybridization among locations provides a rare opportunity for future studies on the consequences of different levels of gene flow for reinforcement, thus informing about the mechanisms underlying the completion of speciation.     

Molecular evidence for the taxonomic status of three species of the <Emphasis Type="Italic">Sphyraena obtusata</Emphasis> group (Perciformes: Sphyraenidae) from East Asia

Genetic differences among the three species of Sphyraena with two gill rakers from East Asia (S. iburiensis, S. obtusata, and S. pinguis, defined recently as the S. obtusata group), were investigated using 799-bp sequences of the mitochondrial cytochrome b gene. Pairwise sequence differences within each of the three species were 0.0–0.4% (S. iburiensis), 0.0–0.4% (S. obtusata), and 0.0–0.6% (S. pinguis), respectively, pairwise sequence differences among the three species being 18.3–18.6% (S. iburiensis vs. S. obtusata), 14.9–15.4% (S. iburiensis vs. S. pinguis), and 17.6–18.3% (S. obtusata vs. S. pinguis), respectively. The extent of the latter were comparable to those among four other congeneric species, S. africana, S. forsteri, S. helleri, and S. japonica (16.0–24.5%). Phylogenetic analysis using the maximum-likelihood method indicated reciprocal monophyly of the three species, each clade being supported by a bootstrap value of 100%. These findings fully supported the taxonomic status of these species, recently elucidated by detailed morphological comparisons. In addition, the S. obtusata group formed a strongly supported clade against the four other congeneric species.