Genetic diversity of a relict plant species, Ligularia sibirica (L.) Cass. (Asteraceae)

Rare plant species can be divided into naturally, ‘old rare’ species and anthropogenically, ‘new rare’ species. Many recent studies explored genetic diversity of ‘new rare’ species. Less is, however, known about genetic diversity of ‘old rare’ species. We examined isozyme genetic variability of 20 populations of an ‘old rare’ plant species, Ligularia sibirica (Asteraceae) in the Czech and Slovak Republic. It is a long-lived perennial herb with mixed-mating breeding system, widely distributed from East Asia to European Russia, with few isolated relict populations in the remaining part of Europe.The results showed high genetic diversity within populations (80.8%) and a low level of genetic differentiation (F_ST = 0.179). Genetic distance between populations correlated significantly with geographic distance. There was also a significant positive correlation between genetic diversity and population size. This is probably caused by destruction of habitats in last centuries and subsequent decrease of population size. Patterns of genetic diversity suggest that the recent distribution is a result of stepwise postglacial migration of the species and subsequent natural fragmentation.We conclude that L. sibirica populations preserve high levels of genetic diversity and are not yet threatened by genetic factors. However, this may change if changes in habitat conditions continue.     

Chemotaxonomy of New Zealand red algae in the family Gigartinaceae (Rhodophyta) based on galactan structures from the tetrasporophyte life-stage

The identification of the polysaccharides from tetrasporophytic plants of nine endemic New Zealand species belonging to the Gigartinaceae, ‘Gigartina’ ancistroclada, ‘G.’ grandifida, Gigartina dilatata, G. divaricata, G. macrocarpa, G. marginifera, G. pachymenioides, G. sp. ‘Lindauer 164’ and Sarcothalia livida using infra-red spectroscopy in conjunction with constituent sugar and glycosyl linkage/substitution analysis is reported. All nine species contain galactans with structures consistent with λ-type carrageenans. Differences in the structures of the galactans in these and a further six previously studied species indicate chemotaxonomically distinct groupings that correspond to Sarcothalia, ‘Sarcothalia’ and Gigartina genera plus some outliers. These distinct, chemotaxonomic groupings are aligned to those determined by rbcL sequence analysis reported in the literature.     

Inter- and intraspecific variation in the genitalia of the ‘Melitaea phoebe group’ (Lepidoptera, Nymphalidae)

The ecology and phylogeny of the Melitaeini butterflies have received considerable attention in the last few years. Several publications have been based on research using molecular methods. Genital morphometry has already been published on two sibling species but without any accompanying taxonomic revision of related species or their intraspecific taxonomy. In this study the morphology of genitalia was analysed by landmark-based geometric morphometrics in males and by traditional morphometrics in females. Based on these morphological characters, the whole M. phoebe group (subg. Cinclidia) was revised. According to our results M. telona Fruhstorfer, 1908 is conspecific with M. ‘phoebe’ ornata Christoph, 1893. Thus, we consider M. telona a junior synonym of M. ornata, and we propose new combinations: M. ornata ornata Christoph, 1893, M. ornata telona Fruhstorfer, 1908, M. ornata amanica Rebel, 1917, M. ornata capreola Varga, 1967, M. ornata emipunica Verity, 1919, M. ornata enoch Higgins, 1941, M. ornata kovacsi Varga, 1967, M. ornata nigrogygia Verity, 1938, M. ornata ogygia Fruhstorfer, 1907 and M. ornata totila Stauder, 1914 based on the rule of priority. The validity of the species status of M. punica and M. scotosia was confirmed, whereas M. sibina and M. pseudosibina could not be separated from M. phoebe using morphometrics. A new species from Iran (Zagros Mts.) was described as M. zagrosi sp. n. In addition, the whole geographical range of M. ornata was reconsidered and some biogeographical connections were discussed. In M. ornata four morphotypes were found based on genitalia: 1. ‘Western type’: including the Italian, Hungarian and SW Ukrainian populations; 2. ‘Eastern type’: with a much extended distribution ranging from the Balkan Peninsula through Asia Minor to Kazakhstan; 3. ‘The Levant type’ (=M. ornata telona comb. nov.); 4. ‘Northern Iranian type’ (Kopeth Dagh and Elburz Mts.). The distribution of these morphotypes can be attributed to historic separation in distinct glacial refugia.     

Where are inion and endinion? Variations of the exo- and endocranial morphology of the occipital bone during hominin evolution

The occipital bone is frequently investigated in paleoanthropological studies because it has several features that help to differentiate various fossil hominin species. Among these features is the separation between inion and endinion, which has been proposed to be an autapomorphic trait in (Asian) Homo erectus. Methodologies are developed here to quantify for the first time the location of these anatomical points, and to interpret their variation due to the complex interactions between exocranial and endocranial size and shape of the occipital and nuchal planes, as well as the occipital lobes and cerebellum. On the basis of our analysis, neither ‘the separation between inion and endinion’ nor ‘endinion below inion’ can be considered as an autapomorphic trait in H. erectus, since this feature is a condition shared by extant African great apes and fossil hominins. Moreover, our results show that the exo- and endocranial anatomy of the occipital bone differs between hominins (except Paranthropus boisei specimens and KNM-ER 1805) and great apes. For example, chimpanzees and bonobos are characterized by a very high position of inion and their occipital bone shows an antero-posterior compression. However, these features are partly correlated with their small size when compared with hominins. Asian H. erectus specimens have a thick occipital torus, but do not differ from other robust specimens, neither in this feature nor in the analysed exo- and endocranial proportions of the occipital bone. Finally, the apparent brain size reduction during the Late Pleistocene and variation between the sexes in anatomically modern humans (AMH) reflect that specimens with smaller brains have a relatively larger posterior height of the cerebellum. However, this trend is not the sole explanation for the ‘vertical shift’ of endinion above inion that appears occasionally and exclusively in AMH.     

Evolution of arid zone birds in Australia: leapfrog distribution patterns and mesic-arid connections in quail-thrush (Cinclosoma, Cinclosomatidae)

The quail-thrush, Cinclosoma, include between five and seven species distributed broadly across arid and semi-arid inland Australia, mesic forests of south-eastern Australia and New Guinea. It has been suggested that the arid zone species of quail-thrush arose from forest ancestors as Australia changed from a warm wet climate to a cooler drier climate since the late-Miocene. We generated multilocus (mitochondrial ND2 and eight nuclear loci) gene and species trees with complete taxon sampling of Cinclosoma to investigate evolutionary relationships and species status of some taxa. Topologies reconstructed in congruent, highly-resolved gene trees and species trees that supported the recognition of seven species. Ancestral state reconstruction and divergence time estimates suggest that arid-adapted taxa radiated in parallel with a drying climate and changing habitat. A ‘leapfrog’ distribution in phenotypes of arid zone taxa was likely a result of ancestral retention of inconspicuous (or camouflaged) plumage patterns. A specimen-based report from 1968 of hybridization between non-sister taxa Cinclosoma castanotum and Cinclosoma marginatum was verified using molecular analysis on specimens collected at the same locality 40 years later. We discuss the implications of hybridization to the evolution of this species group.     

Predator-induced changes in fluted giant clam (Tridacna squamosa) shell morphology

Predator-induced defences have been demonstrated in numerous marine molluscs, but never in giant clams (Bivalvia: Tridacnidae). Water-borne cues from the predatory crab Myomenippe hardwickii were tested for their ability to induce changes in the shell morphology, strength and growth of juvenile fluted giant clams, Tridacna squamosa. Specimens were maintained in effluent from tanks holding ‘fed crabs’, ‘starved crabs’ and ‘no crabs’. To ensure changes were due to predator cues only, food for the crabs was provided ex situ and measures were taken to minimise the prey signal. After 182 days, MANOVA identified differences in various shell parameters relating to shape and strength. CDA showed a clear change in overall morphology, with strong separation among the three treatments. Effluent from ‘fed crabs’ had a greater influence than effluent from ‘starved crabs’, possibly because the starved crabs were perceived as weaker, and thus less risk. Significantly more clams died in ‘fed crabs’ treatments (25%) and ‘starved crabs’ treatments (30%) compared to the ‘no crabs’ control (0%). This degree of mortality has not been observed in similar experiments elsewhere and represents a new challenge to interpret. We suggest that the cause is unlikely to be related to the plastic responses, but rather a result of some form of crab-associated contaminant.     

Sexual dimorphism and polymorphism in a Callovian Phlycticeras (Ammonoidea) assemblage of Kutch, India

An assemblage of Phlycticeras Hyatt from a precisely dated zone of Middle Callovian in Kutch, India has been analyzed. Systematic study reveals that this sample can be divided into two size groups. The larger set shows different adult modifications leading to ornamental polymorphism. Polymorphs are very similar, if not identical, to different chronospecies of Europe, which range between Middle to Upper Callovian. They are grouped under well-known Phlycticeras polygonium (Zieten). The group of smaller specimens on the other hand, strongly resembles a species, which has been previously described as Phlycticeras schaumburgi (Waagen) from the younger Upper Callovian horizons in Kutch. It has a peculiar ‘rooster’-like septicarinate keel in the venter near adult aperture. Phlycticeras-Oecoptychius have been considered as a possible dimorphic pair since long, but here shown to have many inconsistencies to support these views. Instead, sexual dimorphism is explored within Phlycticeras and P. polygonium-P. schaumburgi is proposed as possible antidimorphs. A similar association is found in different stratigraphic assemblages of Europe. Interestingly these two morphs show parallel evolutionary changes within the Phlycticeras genus.     

Himantura tutul sp. nov. (Myliobatoidei: Dasyatidae), a new ocellated whipray from the tropical Indo-West Pacific,described from its cytochrome-oxidase I gene sequence

It has been previously established that the Leopard Whipray, Himantura leoparda, consists of two genetically isolated, cryptic species, provisionally designated as ‘Cluster 1’ and ‘Cluster 4’ (Arlyza et al., Mol. Phylogenet. Evol. 65 (2013) [1]). Here, we show that the two cryptic species differ by the spotting patterns on the dorsal surface of adults: Cluster-4 individuals tend to have larger-ocellated spots, which also more often have a continuous contour than Cluster-1 individuals. We show that H. leoparda's holotype has the typical larger-ocellated spot pattern, designating Cluster 4 as the actual H. leoparda. The other species (Cluster 1) is described as Himantura tutul sp. nov. on the basis of the nucleotide sequence of a 655-base pair fragment of its cytochrome-oxidase I gene (GenBank accession No. JX263335). Nucleotide synapomorphies at this locus clearly distinguish H. tutul sp. nov. from all three other valid species in the H. uarnak species complex, namely H. leoparda, H. uarnak, and H. undulata. H. tutul sp. nov. has a wide distribution in the Indo-West Pacific, from the shores of eastern Africa to the Indo-Malay archipelago. H. leoparda under its new definition has a similarly wide Indo-West Pacific distribution.     

Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)

Despite routine screening requirements for the notifiable fish pathogen Gyrodactylus salaris, no standard operating procedure exists for its rapid identification and discrimination from other species of Gyrodactylus. This study assessed screening and identification efficiencies under real-world conditions for the most commonly employed identification methodologies: visual, morphometric and molecular analyses. Obtained data were used to design a best-practice processing and decision-making protocol allowing rapid specimen throughput and maximal classification accuracy. True specimen identities were established using a consensus from all three identification methods, coupled with the use of host and location information. The most experienced salmonid gyrodactylid expert correctly identified 95.1% of G. salaris specimens. Statistical methods of classification identified 66.7% of the G. salaris, demonstrating the need for much wider training. Molecular techniques (internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP)/cytochrome c oxidase I (COI) sequencing) conducted in the diagnostic laboratory most experienced in the analysis of gyrodactylid material, identified 100% of the true G. salaris specimens. Taking into account causes of potential specimen loss, the probabilities of a specimen being accurately identified were 95%, 87% and 92% for visual, morphometric and molecular techniques, respectively, and the probabilities of correctly identifying a specimen of G. salaris by each method were 81%, 58% and 92%. Inter-analyst agreement for 189 gyrodactylids assessed by all three methods using Fleiss’ Kappa suggested substantial agreement in identification between the methods. During routine surveillance periods when low numbers of specimens are analysed, we recommend that specimens be analysed using the ITS-RFLP approach followed by sequencing of specimens with a “G. salaris-like” (i.e. G. salaris, Gyrodactylus thymalli) banding pattern. During periods of suspected outbreaks, where a high volume of specimens is expected, we recommended that specimens be identified using visual identification, as the fastest processing method, to select “G. salaris-like” specimens, which are subsequently identified by molecular-based techniques.     

Stature estimation from complete long bones in the Middle Pleistocene humans from the Sima de los Huesos, Sierra de Atapuerca (Spain)

Systematic excavations at the site of the Sima de los Huesos (SH) in the Sierra de Atapuerca (Burgos, Spain) have allowed us to reconstruct 27 complete long bones of the human species Homo heidelbergensis. The SH sample is used here, together with a sample of 39 complete Homo neanderthalensis long bones and 17 complete early Homo sapiens (Skhul/Qafzeh) long bones, to compare the stature of these three different human species. Stature is estimated for each bone using race- and sex-independent regression formulae, yielding an average stature for each bone within each taxon. The mean length of each long bone from SH is significantly greater (p < 0.05) than the corresponding mean values in the Neandertal sample. The stature has been calculated for male and female specimens separately, averaging both means to calculate a general mean. This general mean stature for the entire sample of long bones is 163.6 cm for the SH hominins, 160.6 cm for Neandertals and 177.4 cm for early modern humans. Despite some overlap in the ranges of variation, all mean values in the SH sample (whether considering isolated bones, the upper or lower limb, males or females or more complete individuals) are larger than those of Neandertals. Given the strong relationship between long bone length and stature, we conclude that SH hominins represent a slightly taller population or species than the Neandertals. However, compared with living European Mediterranean populations, neither the Sima de los Huesos hominins nor the Neandertals should be considered ‘short’ people. In fact, the average stature within the genus Homo seems to have changed little over the course of the last two million years, since the appearance of Homo ergaster in East Africa. It is only with the emergence of H. sapiens, whose earliest representatives were ‘very tall’, that a significant increase in stature can be documented.     

First record of L-quebrachitol in Allophylus edulis (Sapindaceae)

Allophylus edulis, commonly called ‘Chal chal’, is a member of the Sapindaceae occurring in the Uruguayan and Brazilian native flora. During the phytochemical analysis of two Chal chal specimens from two well-differentiated geographical zones (Assis, São Paulo, Brazil, and Santa Lucía, Canelones, Uruguay), considerable amounts of l-quebrachitol were isolated from both samples. The isolation was carried out from the ethanolic twig extracts obtained by maceration of both vegetal samples. White easily distinguishable crystals were mechanically separated, washed, and characterized by 1D and 2D NMR experiments and by MS data. Such techniques confirmed that the crystals isolated from sources collected in both countries resulted in the same compound, l-quebrachitol, a natural product not previously reported for this species and one that has been investigated as a sugar substitute for diabetics. Worthy of note, the content of l-quebrachitol in A. edulis may be the chemical basis to explain its ethnobotanical uses, since infusions of this plant are used to treat diabetes in the practice of local traditional medicine.     

Gymnocranius superciliosus and Gymnocranius satoi, two new large-eye breams (Sparoidea: Lethrinidae) from the Coral Sea and adjacent regions

Two related perciform fish species of the subfamily Monotaxinae (Sparoidea: Lethrinidae) Gymnocranius superciliosus sp. nov. and Gymnocranius satoi sp. nov. are described from specimens and tissue samples from the Coral Sea and adjacent regions. G. superciliosus sp. nov. is distinct from all other known Gymnocranius spp. by the following combination of characters: body elongated (depth 2.7–3.1 in standard length), caudal fin moderately forked with a subtle middle notch, its lobes slightly convex inside, distinctive blackish eyebrow, snout and cheek with blue speckles, and dorsal, pectoral, anal and caudal fins reddish. G. satoi sp. nov. is the red-finned ‘Gymnocranius sp.’ depicted in previous taxonomic revisions. While colour patterns are similar between the two species, G. satoi sp. nov. is distinct from G. superciliosus sp. nov. by the ratio of standard length to body depth (2.4–2.5 vs. 2.7–3.1) and by the shape of the caudal fin, which is more shallowly forked, its lobes convex inside and their extremities rounded. The two species are genetically distinct from each other and they are genetically distinct from G. elongatus, G. euanus, G. grandoculis, and G. oblongus sampled from the Coral Sea and adjacent regions.     

Tritrichomonas foetus from domestic cats and cattle are genetically distinct

The genetic relationship amongst Tritrichomonas foetus isolated from domestic cats and cattle was investigated by DNA sequencing of the internal transcribed region of the ribosomal DNA unit and the TR7/TR8 variable-length repeat. The results reject the hypothesis that T. foetus from domestic cats is genetically identical to T. foetus in cattle. We suggest recognition of a ‘cat genotype’ and a ‘cattle genotype’ of T. foetus. Review of public nucleotide repositories revealed that the ‘cat genotype’ has not been isolated from cattle nor the ‘cattle genotype’ recovered from cats. However, at least one cat isolate has been shown to induce disease in experimentally infected cattle. We conclude that these genotypes fall within the single species T. foetus.     

Reproductive behaviour of female Chorthippus biguttulus grasshoppers

Female grasshoppers of acoustically communicating species assume series of reproductive states that are associated with particular behaviours. Studies on laboratory populations of Chorthippus biguttulus (L.) revealed that females of this species lack the period of ‘passive copulatory readiness’, increase their attractiveness to males by sound production and mate multiple times before their first oviposition. In particular, female Ch. biguttulus display a period of ‘primary rejection’ after their imaginal moult during which they reject male mating attempts followed by a period of ‘active copulatory readiness’ in which they produce acoustic signals and may copulate with courting males. Female stridulation generally stimulated male mating activity and stridulating females attracted more male mating attempts than mute females in the same cage, indicating that males preferentially court females that signal ‘active copulatory readiness’. After receipt of a spermatophore, Ch. biguttulus females displayed periods of ‘secondary rejection’ followed by re-establishment of ‘active copulatory readiness’. Acoustic responses of females to male songs, an indicator of reproductive readiness, were significantly reduced until 2 days after mating and remained slightly reduced in comparison to pre-mating levels. Some females mated multiple times before their first oviposition and cycled between ‘secondary rejection’ and ‘active copulatory readiness’.     

Sunosuchus (Archosauria, Crocodyliformes) from the Toutunhe Formation (Middle Jurassic) of the Southern Junggar Basin (Xinjiang, NW-China)

A new specimen of the widespread Middle Jurassic to Lower Cretaceous Asian gonipholidid crocodilian genus Sunosuchus is described on the basis of a partial skeleton from the Upper Toutunhe Formation (Middle Jurassic, ?Bathonian-Callovian) of Liuhonggou, SW of Urumqi, Xinjiang Uygur Autonomous Region, People’s Republic of China. The specimen is represented by a partial mandible, teeth, vertebrae, limb and girdle bones and osteoderms. It can be distinguished from other nominal species of the genus by a unique combination of characters: slightly heterodontous dentition, strongly sculptured posteroventral part of the mandible, short fenestra mandibularis, convex dorsal surface of the retroarticular process, keeled cervical vertebral centra and ventral osteoderms with a distinctive sculpture of wide pits and narrow ridges. The heterodontous dentition is a potential autapomorphy of this form. The new specimen is closest in morphology to material described recently from the Callovian of Kirghisia as Sunosuchus sp. It represents the second Middle Jurassic record of the genus, the first crocodile from the Toutunhe Formation, the first substantial crocodile find from the Mesozoic of the Southern Junggar Basin, and the first Middle Jurassic record of Sunosuchus from China. This extends both the paleobiogeographical distribution of the genus in Asia and its stratigraphic distribution in China considerably.     

Description of a new bat species of the tribe Scotonycterini (Chiroptera,Pteropodidae) from southwestern Cameroon

The tribe Scotonycterini is currently composed of three fruit bat species of the family Pteropodidae (Mammalia, Chiroptera) characterized by white fur patches on the head, specifically around the nose and behind the eyes: Scotonycteris zenkeri, S. ophiodon and Casinycteris argynnis. Herein a new species is described, Casinycteris campomaanensis sp. nov., based on female specimen collected in 2007 near the village Nkoélon-Mvini close to the Campo-Ma’an National Park, southwestern Cameroon. It is readily distinguished from the three other species of Scotonycterini by its body size and craniodental characteristics. Molecular analyses based on the complete mitochondrial cytochrome b gene indicate that the new species is the sister-group to C. argynnis and that the holotype of S. ophiodon is more closely related to Casinycteris than to S. zenkeri, rendering the genus Scotonycteris paraphyletic. Based on these results, morphological characters within the tribe Scotonycterini were reassessed and a new classification is proposed, in which the new species and S. ophiodon are placed in the genus Casinycteris.     

Insight into the role of cetaceans in the life cycle of the tetraphyllideans (Platyhelminthes: Cestoda)

Four types of tetraphyllidean larvae infect cetaceans worldwide: two plerocercoids differing in size, ‘small’ (SP) and ‘large’ (LP), and two merocercoids referred to as Phyllobothrium delphini and Monorygma grimaldii. The latter merocercoid larvae parasitize marine mammals exclusively and exhibit a specialised cystic structure. Adult stages are unknown for any of the larvae and thus the role of cetaceans in the life cycle of these species has been a long-standing problem. The SP and LP forms are thought to be earlier stages of P. delphini and M. grimaldii that are presumed to infect large pelagic sharks that feed on cetaceans. A molecular analysis of the D2 variable region of the large subunit ribosomal DNA gene based on several individuals of each larval type collected from three Mediterranean species of cetaceans showed consistent and unique molecular signatures for each type regardless of host species or site of infection. The degree of divergence suggested that LP, P. delphini and M. grimaldii larvae may represent separate species, whereas SP may be conspecific with M. grimaldii. In all host species, individuals of SP accumulated in the gut areas in which the lymphoid tissue was especially developed. We suggest therefore that these larvae use the lymphatic system to migrate to the abdominal peritoneum and mesenteries where they develop into forms recognizable as M. grimaldii. The plerocercoid stage of P. delphini remains unknown. In a partial phylogenetic tree of the Tetraphyllidea, all larvae formed a clade that included a representative of the genus Clistobothrium, some species of which parasitize sharks such as the great white which is known to feed on cetaceans. A bibliographic examination of tetraphyllidean infections in marine mammals indicated that these larvae are acquired mostly offshore. In summary, the evidence suggests that cetaceans play a significant role in the life cycle of these larvae. In addition, it seems clear that cetaceans act as natural intermediate hosts for P. delphini and M. grimaldii, as within these hosts they undergo development from the plerocercoid stage to the merocercoid stage. Because tetraphyllidean species use fish, cephalopods and other marine invertebrates as intermediate hosts, the inclusion of cetaceans in the life cycle would have facilitated their transmission to apex predators such as the large, lamnid sharks. The biological significance of infections of LP in cetaceans is unclear, but infections do not seem to be accidental as such larvae show high prevalence and abundance as well as a high degree of site specificity, particularly in the anal crypts and bile ducts.