Introgression and species demarcation in western European Leuctra fusca (Linnaeus, 1758) and L. digitata Kempny, 1899 (Plecoptera: Leuctridae)

The compilation of a DNA barcoding library of Norwegian stonefly (Plecoptera) species revealed that Leuctra fusca (Linnaeus, 1758 Linnaeus, C. (1758), Systema naturae Per Regna Tria Naturae: Secundum Classes, Ordines, Genera, Species, Cum Characteribus, Differentiis, Synonymis, Locis. Stockholm: Laurentius Salvius. [Google Scholar]) and Leuctra digitata Kempny, 1899 (Leuctridae) share haplotypes in northernmost Scandinavia. Phylogenetic analyses of the mitochondrial (mt) DNA barcode marker COI and the nuclear marker 28S show that the shared haplotypes must result from the introgression of a L. fusca mitochondrion into a L. digitata population on at least two occasions. Although mt introgression is widespread in animals, this represents the first documented case in Plecoptera. This study also included specimens of L. cf. fusca from the Sierra Nevada massif in Spain, a population previously known as L. carpentieri Despax, 1945. Their mt haplotypes are ca. 13% different from other European L. fusca. However, their 28S alleles are compatible with their morphological identification as L. fusca. In view of the possibility of mt introgression, the taxonomic status of this population remains undecided.     

Dispersal,feeding and parasitism of adult stoneflies (Plecoptera) at a New Zealand forest stream

The temporal occurrence and dispersal of stoneflies were investigated at a forest stream in the South Island, New Zealand. Although 12 species were taken in Malaise traps, only Spaniocerca zelandica and Cristaperla fimbria (Notonemouridae) were abundant. Adults of S. zelandica were present throughout the year and a wide range of different-sized nymphs, including very small individuals, was also found in all months. In contrast, the emergence period of C. fimbria was limited to about four months in summer and as nymphs are mainly hyporheic, few were taken in Surber samples. Malaise trapping on a 16 m long transect away from the stream and with more distantly located traps in forest and grassland resulted in a very high proportion of males and females of both species being caught within 1 m of the stream edge. Captured females of both species included immature, gravid, and spent individuals. Males and females of both species had almost identical diets dominated by sooty mould fungi, fungal spores, pollen and fine particulate organic matter. High proportions of them were also infected by the encysted larvae of a gordian worm (Nematomorpha) and may be important vectors in its life cycle.     

The adult diet of <Emphasis Type="Italic">Xanthoperla apicalis</Emphasis> and <Emphasis Type="Italic">Siphonoperla torrentium</Emphasis> (Plecoptera,Chloroperlidae) in the Danube basin (Slovakia)

The gut contents of adult Xanthoperla apicalis and Siphonoperla torrentium were analysed. X. apicalis females eat mainly pollen, while males feed principally on detritus and pollen. Adults of S. torrentium, regardless of sex, eat mainly pollen. The presence of animal remains (claws and leg and antennae segments of Arthropoda) in the gut of several individuals of both species was observed and discussed.     

Genetic and phenotypic variability in Stenoperla prasina (Newman, 1845) (Plecoptera: Eustheniidae) in relation to latitude and altitude in New Zealand

Genetic divergence, body size and variations in external genitalia of the stonefly Stenoperla prasina (Newman, 1845) (Plecoptera: Eustheniidae) were investigated using specimens from 46 locations throughout New Zealand. Sequencing of a 658-bp fragment of the mtCOI gene from 77 individuals collected at 26 locations identified 10 haplotypes, and three geographic haplotype networks. Maximum uncorrected genetic divergence found was 4.3%. Past isolation of populations in northern and southwestern New Zealand was suggested by variations in genitalia. No relationship was found between body size and latitude over the length of New Zealand. However, male or female size was correlated with either latitude or altitude in the North or South Islands, and with altitude on a North Island mountain. The relatively small number of haplotypes found could indicate that they are survivors of an event that caused a bottleneck.     

Spermatozoa morphology and ultrastructure in Nile perch,Lates niloticus (Linnaeus, 1758)

Lates niloticus is a valuable commercial fish species with good potential for aquaculture. However, there is limited information on the type and structure of the Nile perch spermatozoon, which could potentially aid in culture of this species. Here, we describe the spermatozoon ultrastructure in L. niloticus using transmission and scanning electron microscopy. The spermatozoon had a round head-shape, medio-laterally flat, no acrosome, a short midpiece located laterally to the nucleus, uniflagella with one wing. The head of the spermatozoon contained the nucleus, centriolar system, proximal part of the flagellum, and cytoplasmic channel. Centrioles were arranged at an angle of 90° to each other, forming a T-shape, parallel to the nucleus. The midpiece was cylindrical, loaded with cytoplasm, five to seven spherical mitochondria; and the flagellum’s plasma membrane extended to form one lateral wing. The spermatozoa were classified as type II spermatozoa. L. niloticus spermatozoon differed from that of its Australian congener L. calcarifer, especially in the centriole arrangement and nuclear shape, length of the midpiece and the number of mitochondria and lateral wings.     

Genetic structure of house mouse (Mus musculus Linnaeus 1758) populations in the Atlantic archipelago of the Azores: colonization and dispersal

We analyzed the genetic structure and relationships of house mouse (Mus musculus) populations in the remote Atlantic archipelago of the Azores using nuclear sequences and microsatellites. We typed Btk and Zfy2 to confirm that the subspecies Mus musculus domesticus was the predominant genome in the archipelago. Nineteen microsatellite loci (one per autosome) were typed in a total of 380 individuals from all nine Azorean islands, the neighbouring Madeiran archipelago (Madeira and Porto Santo islands), and mainland Portugal. Levels of heterozygosity were high on the islands, arguing against population bottlenecking. The Azorean house mouse populations were differentiated from the Portuguese and Madeiran populations and no evidence of recent migration between the three was obtained. Within the Azores, the Eastern, Western, and Central island groups tended to act as separate genetic units for house mice, with some exceptions. In particular, there was evidence of recent migration events among islands of the Central island group, whose populations were relatively undifferentiated. Santa Maria had genetically distinctive mice, which may relate to its colonization history. © 2013 The Linnean Society of London     

Squamation and scale character stability in cichlids, examined in Sarotheroden galilaeus (Linnaeus, 1758) (Perciformes, Cichlidae)

Squamation and scale morphology provide interesting characters for phyletic analysis within the Cichlidae. It is not known, however, to what extent these characters are genetically fixed or determined by environmental parameters. To clarify this point, a study on Sarotherodon galilaeus was conducted. This species has an extensive distribution and lives in markedly different environments. Most scale characters were found to be remarkably stable within the species. The only large variations occur in scale counts, extent of caudal fin Squamation, number of circuli and radii, and the extent of scale granulation. The Squamation pattern, the form of the circuli and radii, the shape of interradial rostrad projecting tongues, the denticles on the circuli in the rostral field, and especially the kind of granulation in the caudal field is similar in all specimens examined and is also retained in fish bred in aquaria for several generations. Thus it seems justified to assume that these characters are strongly determined by genetic factors and that their analysis can be useful in the determination of phyletic relationships.     

Development and ultrastructure of the thickened serosa and serosal cuticle formed beneath the embryo in the stonefly Scopura montana Maruyama, 1987 (Insecta,Plecoptera, Scopuridae)

We aimed to describe the development and ultrastructure of the thickened serosa and serosal cuticle formed beneath the embryo of Plecoptera, using Scopura montana of Scopuridae as a euholognathan representative. Using transmission electron microscopy, we found that the egg membranes were composed of a thick exochorion, a thicker endochorion consisting of two sublayers, and an extremely thin vitelline membrane. The egg membrane construction represents a groundplan feature of the euholognathan egg membranes. The serosa converges beneath the embryo to form a thickened serosa, comprising cells in a radial arrangement, in association with the formation of the amnioserosal fold. The thickened serosa then deposits the thickened serosal cuticle, consisting of four layers differing in fine structure and electron density. After achieving its secretory function, the thickened serosa then disintegrates, and the liberated serosal cells float for a short period in the peripheral region of the egg inside. Collectively, our findings should provide the basis for further characterization of the serosal structures concerned, but we were unable to corroborate previous studies assigning the thickened serosa and serosal cuticle in Plecoptera to the water absorption function.     

Live cycles,drift, and standing stocks of some stoneflies (Insecta: Plecoptera) from streams in Minnesota,U.S.A.

From analyses of monthly length frequency distributions of stonefly (Plecoptera) nymphs from stream riffles, two types of univoltine life cycles (Hynes 1961, classification) were observed: slow seasonal forIsoperla transmarina, Isoperla signata, Isogenoides olivaceus, andAllocapnia rickerii, and fast seasonal forProstoia similis. The Perlodidae stoneflies (Isoperla andIsogenoides) showed greatest propensities to drift near the end of their life cycles. Drift rates of the other two species of stoneflies were correlated to standing stock estimates.     

Morphology and chemical composition of metathoracic scent glands in Dolycoris baccarum (Linnaeus, 1758) (Heteroptera: Pentatomidae)

One of the general defining characters of the Heteroptera is the presence of metathoracic scent glands (MTG). Using scanning electron microscopy, the morphology of the MTG of Dolycoris baccarum (Linnaeus 1758) (Heteroptera: Pentatomidae) was studied. The MTG belong to the diastomian type. The two glandular pores located between the mesothoracic and metathoracic coxae are associated with 'mushroom-like' structures. The MTG are composed of a reservoir and a pair of lateral glands is connected to the reservoir by a duct. A groove-like structure extends downwards from the ostiole. While this structure is long and wide, its ostiole is oval. Extracts of the volatile fractions from male and female MTG secretions were analysed by capillary gas chromatography–mass spectrometry (GC-MS) and exhibited a typical pentatomid composition. Seventeen chemical compounds were detected in female secretions, whereas 13 chemical compounds were detected in the male secretions. Most chemical compounds were similar between the sexes but were different in their quantities. In this regard, the compounds identified were investigated and the biological functions of the glandular secretions were discussed. In the analyses of the MTG of females of D. baccarum , tridecane (50.97%) was a major odour component and (Z,Z)-4,16-octadecadien-1-ol acetate (0.02%) was a minor odour component. In males, tridecane (50.80%) was a major odour component and 1,2-benzenedicarboxylic acid (0.02%) was a minor odour component.     

Microsatellite polymorphism and population subdivision in natural populations of European sea bass Dicentrarchus labrax (Linnaeus, 1758)

Polymorphism of microsatellite markers was used to study the genetic variability and structure in natural populations of European sea bass Dicentrarchus labrax. The data consisted of six microsatellite loci analysed for 172 individuals from three samples collected in the Golfe-du-Lion (France) and one sample collected in the Golfo-de-Valencia (Spain). Our goals were (i) to assess the level of genetic variability as revealed by these markers, (ii) to estimate the genetic differentiation among natural populations within a restricted area, and (iii) to evaluate how microsatellite loci fit the predictions of the two most widely used mutation models (the infinite allele model and the stepwise mutation model). As expected, our results indicate that the genetic polymorphism is very high when compared with previously used genetic markers, the mean expected heterozygosity per locus ranging between 0.69 and 0.93. We also found that all loci but one fitted the infinite allele model better. Using this model as a lower limit, we could extrapolate from the observed diversity effective population sizes on the order of 35 000 individuals. Our results also suggest that there may be a slight genetic differentiation between the two gulfs (F_ST= 0.007, P < 0.05), indicating that the corresponding populations are likely to be dynamically independent. This finding for a species with high dispersal abilities, if confirmed, has important beatings on fish-stock assessment.     

Temporal scales of variation in settlement and recruitment of the mussel Perna perna (Linnaeus, 1758)

Population dynamics of many intertidal organisms are strongly affected by the abundance and distribution of larvae arriving on the shore. In particular, not only absolute numbers of settlers but also the degree of synchronisation of settlement can have a strong influence on whether density-dependent or density-independent processes shape adult shape populations. Temporal variation in rates of settlement and recruitment of the mussel Perna perna on the south coast of South Africa was investigated using a nested spatial design at different temporal scales. Variability in settlement at spring tides was examined at two temporal scales: lunar (to investigate the effect of state of the moon on settlement) and tidal (to investigate the influence of state of the tide on mussel settlement). Recruitment over neap tides was examined at one temporal scale, fortnight (to investigate the effect of date on mussel recruitment).Strong temporal variation was evident for both settlement and recruitment, but not at all time scales. Distinct peaks of settler/recruit abundance were observed during the lunar and neap tide studies. Recruitment intensity differed over the course of the year, and pulsing of recruitment was generally synchronised among locations. However, the strength of pulsing differed dramatically among locations, giving a significant interaction between fortnight and location. The finest temporal scale, investigated in the tidal study, did not reveal a significant effect of the state of the tide on settlement. The state of the moon (new or full) was not significant as a main factor (p = 0.052), although generally more settlers arrived on the shore during new moon. Phase of the moon appeared to have an effect on settler abundances, but only when and where densities were high.     

Feeding ecology and trophic level of Chimaera monstrosa Linnaeus, 1758 (Holocephali: Chimaeridae) in the Eastern Mediterranean

The feeding ecology of Chimaera monstrosa at a depth of 550?m was studied off S??ac?k Bay, Eastern Mediterranean, in 2008. Forty males and 57 females with a length between 7.80 and 45.5?cm were caught. 33 stomachs contained food items. The diet was composed of 11 prey items belonging to 6 groups: Bryozoa, Cnidaria, Crustacea, Mollusca, Tunicata, and Teleostei, of which Crustacea (IRI% [Index of Relative Importance]=80) and Mollusca (IR%=16) were the most abundant prey groups. The trophic level was estimated at 3.50, which is similar to the results from other studies in the western and northern part of the Mediterranean and the Atlantic Ocean but is higher than in the North Sea.     

Genetic divergence in the Atlantic-Mediterranean Montagu's blenny, Coryphoblennius galerita (Linnaeus 1758) revealed by molecular and morphological characters

Coryphoblennius galerita is a small intertidal fish with a wide distribution and limited dispersal ability, occurring in the northeastern Atlantic and Mediterranean. In this study, we examined Atlantic and Mediterranean populations of C. galerita to assess levels of genetic divergence across populations and to elucidate historical and contemporary factors underlying the distribution of the genetic variability. We analyse three mitochondrial and one nuclear marker and 18 morphological measurements. The combined dataset clearly supports the existence of two groups of C. galerita: one in the Mediterranean and another in the northeastern Atlantic. The latter group is subdivided in two subgroups: Azores and the remaining northeastern Atlantic locations. Divergence between the Atlantic and the Mediterranean can be the result of historical isolation between the populations of the two basins during the Pleistocene glaciations. Present-day barriers such as the Gibraltar Strait or the 'Almeria-Oran jet' are also suggested as responsible for this isolation. Our results show no signs of local extinctions during the Pleistocene glaciations, namely at the Azores, and contrast with the biogeographical pattern that has been observed for Atlantic-Mediterranean warm-water species, in which two groups of populations exist, one including the Mediterranean and the Atlantic coast of western Europe, and another encompassing the western tropical coast of Africa and the Atlantic islands of the Azores, Madeira and Canaries. Species like C. galerita that tolerate cooler waters, may have persisted during the Pleistocene glaciations in moderately affected locations, thus being able to accumulate genetic differences in the more isolated locations such as the Azores and the Mediterranean. This study is one of the first to combine morphological and molecular markers (mitochondrial and nuclear) with variable rates of molecular evolution to the study of the relationships of the Atlantic and Mediterranean populations of a cool-water species.     

The combined effect of temperature and salinity changes on osmoregulation and haemocyanin concentration in Saduria entomon (Linnaeus, 1758)

Global warming is having an impact on the temperature and salinity of Baltic Sea waters. Therefore, it is important to determine the conditions in which animals can exist and how these changes may influence their functioning. Hence, the purpose of this research was to determine the broad tolerance limits of temperature and salinity of the glacial relict Saduria entomon by studying its behaviour, osmoregulatory ability and haemocyanin concentration. This effect of temperature was confirmed in the laboratory for individuals acclimated to different salinity and temperature regimes. Changes in the physiological parameters of S. entomon at various temperatures (5.5–21.5°C) and salinity levels (1–15 PSU) were recorded. There were statistically significant differences in haemolymph osmotic pressure under the influence of salinity and temperature. The mean haemolymph osmotic pressures were the lowest at 1 PSU at all the temperatures examined and the highest at 15 PSU and high temperatures 16.5 and 21.5°C. The haemocyanin concentration decreased significantly with increasing temperature at 1 PSU. There was a significant difference in haemocyanin concentration due to salinity at temperatures of 5.5 and 10.0°C (the haemocyanin concentration decreased with increasing salinity). The results showed that, although S. entomon is classified as a cold-water animal, it can survive at high temperatures above 16.5°C at least for a short time, as it is capable of osmoregulation. The tolerance to temperature changes was better than expected.     

Condition and size of the non‐native pikeperch Sander lucioperca (Linnaeus, 1758) in Portuguese river basins

We studied life‐history traits focusing on the growth and condition of the pikeperch Sander lucioperca to evaluate its phenotypic plasticity when introduced to new environments. Pikeperch is a non‐native fish introduced to Iberian freshwater fauna in 1998 that quickly spread to other river basins through human‐mediated activities, occupying now a wide variety of habitats along mainland Portugal. Condition (K and SMI), fork length at age, and length–weight relationships were studied for Portuguese populations. Pikeperch fork length for ages 1, 2, 3, and 4 was different between several populations. We applied generalized linear models (GLM) to study the influence of habitat type, latitude, altitude, time after first detection, and fish prey richness on pikeperch populations size at age 4 and condition. We observed higher condition values on populations from lower altitudes at lentic systems more recently introduced. But higher fork length at age 4 was found in populations from higher altitudes, on older populations with higher prey richness. Habitat type, time since first detection, and fish fauna composition are discussed as the main environmental factors explaining the observed phenotypic plasticity with concerns on predatory impact on native fauna.     

Anatomy of the tongue and microstructure of the lingual papillae in the fallow deer Dama dama (Linnaeus, 1758)

The function of mouth organs in ruminants is connected with the process of rumination. To study morphofunctional relations, microstructures in tongues of 4- to 5-year old adult fallow deer were examined using scanning electron microscopy.When analyzing the tongue of the fallow deer, i.e. a ruminant classified as an intermediate mixed feeder between grass and roughage eaters, two processes were taken into account: (i) foraging and forage selecting, and (ii) chewing the cud during rumination to reduce particle size and improve digestibility.Microstructural results show that the above mentioned processes in fallow deer are important selection factors, which in the anterior part of tongue led to the development of clusters of fungiform papillae connected with preselection of food as well as a specific pattern of filiform papillae promoting increased adhesion of transported food. Massive and flattened conical papillae on the torus are arranged according to sideways jaw movements and are co-localized with flattened fungiform papillae and two rows of vallate papillae. Such an arrangement of papillae on the lingual torus presumably facilities distribution of ruminated food, with simultaneous transferring of taste signals about masticated food particles.     

Evidence for cryptic speciation in Carchesium polypinum Linnaeus, 1758 (Ciliophora: Peritrichia) inferred from mitochondrial, nuclear, and morphological markers

Protist diversity is currently a much debated issue in eukaryotic microbiology. Recent evidence suggests that morphological and genetic diversity might be decoupled in some groups of protists, including ciliates, and that these organisms might be much more diverse than their morphology implies. We sought to assess the genetic and morphological diversity of Carchesium polypinum, a widely distributed peritrich ciliate. The mitochondrial marker cytochrome c oxidase subunit I and the nuclear small subunit ribosomal RNA were used to examine genetic diversity. For the morphological assessment, live microscopy and Protargol staining were used. The mitochondrial marker revealed six robust, deeply diverging, and strongly supported clades, while the nuclear gene was congruent for three of these clades. There were no major differences among individuals from the different clades in any of the morphological features examined. Thus, the underlying genetic diversity in C. polypinum is greater than what its morphology suggests, indicating that morphology and genetics are not congruent in this organism. Furthermore, because the clades identified by the mitochondrial marker are so genetically diverse and are confirmed by a conserved nuclear marker in at least three cases, we propose that C. polypinum be designated as a "cryptic species complex." Our results provide another example where species diversity can be underestimated in microbial eukaryotes when using only morphological criteria to estimate species richness.