A new fast real‐time PCR method for the identification of three sibling Apodemus species (A. sylvaticus,A. flavicollis,and A. alpicola) in Italy

The identification of field mice Apodemus flavicollis, Apodemus sylvaticus, and Apodemus alpicola represents a challenge for field scientists due to their highly overlapping morphological traits and habitats. Here, we propose a new fast real‐time PCR method to discriminate the three species by species‐specific TaqMan assays. Primers and probes were designed based on the alignment of 54 cyt‐b partial sequences from 25 different European countries retrieved from GenBank. TaqMan assays were then tested on 133 samples from three different areas of Italy. Real‐time PCR analysis showed 92 samples classified as A. flavicollis, 13 as A. sylvaticus, and 28 as A. alpicola. We did not observe any double amplification and DNA sequencing confirmed species assignment obtained by the TaqMan assays. The method is implementable on different matrices (ear tissues, tail, and blood). It can be used on dead specimens or on alive animals with minimally invasive sampling, and given the high sensitivity, the assay may be also suitable for degraded or low‐DNA samples. The method proved to work well to discriminate between the species analyzed. Furthermore, it gives clear results (amplified or not) and it does not require any postamplification handling of PCR product, reducing the time needed for the analyses and the risk of carryover contamination. It therefore represents a valuable tool for field ecologists, conservationists, and epidemiologists.     

A novel approach to field identification of cryptic Apodemus wood mice: calls differ more than morphology

Field identification of European wood mice Apodemus spp. is challenging due to their morphological resemblance and frequent sympatry. We developed discriminant functions based on body mass and acoustic variables of distress calls to identify three cryptic species of wood mice (Apodemus alpicola, Apodemus flavicollis and Apodemus sylvaticus) in Italy. We achieved an overall correct classification rate of 86–98%; the best results (100% correct classification) were obtained for Apodemus sylvaticus calls. This minimally invasive, effective and low‐cost method highlights the potential role of bioacoustics as a powerful tool for field discrimination of cryptic species of terrestrial mammals.     

Changes in home range size during growth and maturation of the wood mouse (Apodemus sylvaticus) and the bank vole (Clethrionomys glareolus)

Summary A field study was carried out during winter and spring of 1982/83 to determine changes in home range size with increasing body weight and maturation in the wood mouse (Apodemus sylvaticus) and the bank vole (Clethrionomys glareolus). Individual home range sizes in males increased and in females decreased with increasing body weight. Relating these changes to changed energetic requirements failed. In these species behavioral aspects seem to be a more important determinant of home range size than body weight.     

Diversity of fungus‐growing termites (Macrotermes) and their fungal symbionts (Termitomyces) in the semiarid Tsavo Ecosystem,Kenya

Fungus‐growing termites of the subfamily Macrotermitinae together with their highly specialized fungal symbionts (Termitomyces) are primary decomposers of dead plant matter in many African savanna ecosystems. The termites provide crucial ecosystem services also by modifying soil properties, translocating nutrients, and as important drivers of plant succession. Despite their obvious ecological importance, many basic features in the biology of fungus‐growing termites and especially their fungal symbionts remain poorly known, and no studies have so far focused on possible habitat‐level differences in symbiont diversity across heterogeneous landscapes. We studied the species identities of Macrotermes termites and their Termitomyces symbionts by excavating 143 termite mounds at eight study sites in the semiarid Tsavo Ecosystem of southern Kenya. Reference specimens were identified by sequencing the COI region from termites and the ITS region from symbiotic fungi. The results demonstrate that the regional Macrotermes community in Tsavo includes two sympatric species (M. subhyalinus and M. michaelseni) which cultivate and largely share three species of Termitomyces symbionts. A single species of fungus is always found in each termite mound, but even closely adjacent colonies of the same termite species often house evolutionarily divergent fungi. The species identities of both partners vary markedly between sites, suggesting hitherto unknown differences in their ecological requirements. It is apparent that both habitat heterogeneity and disturbance history can influence the regional distribution patterns of both partners in symbiosis.     

Complex mutation at a microsatellite locus in sturgeons: Acipenser sinensis, A. schrenckii, A. gueldenstaedtii and A. baerii

Cross‐species amplifications of microsatellite locus Spl‐106, which was originally screened from the genome of shovelnose sturgeon (Scaphirhynchus platorynchus) with a perfect TAGA repeat motif, were carried out in four other species of the genera Acipenser. A total of 34 polymerase chain reaction (PCR) products representing 16 different alleles of this locus was sequenced. Sequence analysis results showed that besides the number changes of repeat units, many mutational events, such as single‐base substitutions and various insertion/deletion (indels) occurred not only at species level but also at individual level, even among the different alleles within the same individual. The repeat motifs varied from perfect (TAGA)n array to perfect compound (TAAA)m (GAAA)n and perfect or imperfect compound (TAAA)m (TAGA)n (TAAA)x arrays in different species and different individuals. The evolution dynamics of this locus in sturgeons was inferred in that it may evolve from a single perfect to different perfect or imperfect compounds.     

A NEW AGAROPHYTE SPECIES,GELIDIUM EUCORNEUM SP. NOV. (GELIDIALES,RHODOPHYTA), BASED ON MOLECULAR AND MORPHOLOGICAL DATA1

Gelidium is an economically and ecologically important agar‐producing genus. Although the taxonomy of Gelidium has been the focus of many published studies, there is still a need to reevaluate species‐level diversity. Herein, we describe Gelidium eucorneum sp. nov. based on specimens collected off Geojedo on the southern coast of Korea. G. eucorneum is distinguished by cartilaginous thalli with brush‐like haptera, rhizoidal filaments concentrated in the medulla, and globose cytocarps that are horned with multiple determinate branchlets. The species occurs in wave‐exposed intertidal sites, sometimes in association with other mat‐forming algae. Phylogenetic analyses (rbcL, psaA, and cox1) reveal that G. eucorneum is unique and clearly distinct from other species of the genus. The clade containing Gelidium vagum and Acanthopeltis longiramulosa was resolved as a sister group to G. eucorneum. We suggest that the diverse morphologies of G. eucorneum, G. vagum, and Acanthopeltis developed from a common ancestor in East Asian waters. This study shows that even in well‐studied areas, more agarophyte species are to be added to the world inventory of red algae.     

A removal experiment with sympatric populations of Apodemus sylvaticus (L.) and A. flavicollis (Melchior) (Rodentia: Muridae)

Summary This study examines the role of competition in the relationship between sympatric populations of Apodemus sylvaticus and A. flavicollis at Woodchester Park, Gloucestershire, England. Three grids were trapped for twelve months to monitor changes in population size, survival, reproduction, body weight, spatial segregation and microhabitat associations. The A. flavicollis was removed from one grid, A. sylvaticus from another while the rodent community on the third was left intact as a control. Subsequent changes in ecological parameters were observed over a further period of 7 months. Isolated populations of both species entered reproductive condition earlier than on the control grid. This resulted in a rapid increase of the A. flavicollis population but the dynamics of A. sylvaticus were unaffected. In the absence of their congener both species made greater use of areas of woodland once frequented by the absent species. However specific microhabitat associations observed in two species communities remained after experimental manipulation. It is concluded that although A. sylvaticus and A. flavicollis do compete for space, this is a comparatively weak interaction which does not lead to habitat exclusion. Competition is probably ameliorated by intrinsic differences in the biology of these Apodemus species.     

Ecology of Hantaan virus at Twin Bridges Training Area,Gyeonggi Province,Republic of Korea, 2005–2007

The Twin Bridges Training Area (TBTA) in the Republic of Korea consists of dirt roads, barren training areas, and forested hillsides adjacent to linear and broad expanses of tall grasses, herbaceous, and scrub vegetation. Of the six species of small mammals, the striped field mouse, Apodemus agrarius, was the most frequently captured (96.1%). Apodemus agrarius capture rates varied from 17.7 to 33.2% during three trapping periods. Gravid females were observed during November‐December 2006 (8.4%) and March 2007 (5.1%). In 2005, the overall seroprevalence of Hantaan virus (HTNV) was high (34.4%) and lower during surveys in 2006 (14.2%) and 2007 (13.8%). Seroprevalence was directly correlated with weight increase of A. agrarius.     

Geographic distribution and abundance of woolly (Lagothrix cana) and spider (Ateles chamek) monkeys in southwestern Brazilian Amazonia.

Two species of frugivorous atelids, Ateles chamek and Lagothrix cana, occur in southwestern Brazilian Amazonia. Populations were surveyed at 36 sites in the state of Rondônia. Ateles chamek is widespread, but the distribution of L. cana is limited by a combination of riverine barriers and ecological factors, possibly including competition with A. chamek. Groups of L. cana were generally larger and more abundant than those of A. chamek, even in syntopy. The transitional forest that predominates in the extreme south of Rondônia (Hylea‐cerrado) is not a barrier to either species, with both species being tolerant of habitat disturbance when hunting pressure is low. Am. J. Primatol. 56:57–64, 2002. © 2002 Wiley‐Liss, Inc.     

Effects of acorn masting on population dynamics of three forest-dwelling rodent species in Hokkaido,Japan

The effects of the abundance of acorns of the oak, Quercus crispula, on the population dynamics of three rodent species (Apodemus speciosus, A. argenteus, and Clethrionomys rufocanus) were analyzed using time series data (1992–2006). The data were obtained in a forest in northern Hokkaido, Japan, by live trapping rodents and directly counting acorns on the ground. Apodemus speciosus generally increased in abundance following acorn masting. However, the clear effect of acorn abundance was not detected for the other two rodent species. Acorns of Q. crispula contain tannins, which potentially have detrimental effects on herbivores. Apodemus speciosus may reduce the damage caused by acorn tannins with tannin-binding salivary proteins and tannase-producing bacteria, whereas such physiological tolerance to tannins is not known in the other two rodent species. The differences in the effects of acorns between the three species may be due to differences in their physiological tolerance to tannins.     

Field trials of slow‐release synthetic lures for stoats (Mustela erminea) and ferrets (M. furo)

Abstract

Experimental field trials were used to compare the attractiveness to stoats (Mustela erminea) and ferrets (M. furo) of traps set with either a synthetic scent lure or with fresh food bait. One lure, containing 2‐n‐propylthietane, effectively attracted as many ferrets as did rabbit meat bait. Fewer stoats were caught with this lure than with egg bait. Made up in a slow‐release formulation in “plastic rope”, the lure remained attractive to ferrets for at least 2 weeks under sunny field conditions. Traps containing a second lure, 3‐n‐propyl‐l,2‐dithiolane, were less successful at catching both mustelid species than those containing fresh food baits. The scent lures were not attractive to non‐target species, but neither did they deter some species.     

Patterns of intraspecific variation inAntennaria alborosea,A. corymbosa,A. marginata,A. microphylla,A. parvifolia,andA. umbrinella (Asteraceae:Inuleae)

Patterns of intraspecific variation were examined inAntennaria alborosea A. E. Porsild,A. corymbosa E. Nels,A. marginata Greene,A. microphylla Rydb.,A. parvifolia Nutt., andA. umbrinella Rydb. AlthoughA. alborosea was initially considered arctic in distribution, it became apparent that a southern montane element also exists. Our results suggest that morphological differences between arctic and southern montane specimens represent clinal variation. The additional morphological data for specimens that occur more than 1,500 km south of the species' range as it was initially described result in a better understanding of this once presumed arctic taxon. Morphological variation in the dioecious speciesA. corymbosa, A. marginata, A. microphylla, A. parvifolia, andA. umbrinella was greater between the genders than was geographic variation within each gender. These results demonstrate that both pistillate and staminate specimens must be examined in dioecious species ofAntennaria if morphological variation in the respective species is to be fully understood. Character size or number of broadly distributed species (A. microphylla andA. parvifolia) generally decreased with increasing longitude, whereas characters of species with more restricted distributions (A. alborosea, A. corymbosa, andA. marginata) generally increased in size or number with increasing latitude or longitude.Antennaria umbrinella was an exception in this respect.     

A taxonomic revision of a Hypnea charoides-valentiae complex (Rhodophyta, Gigartinales) in Japan, with a description of Hypnea flexicaulis sp. nov.

Morphology of a Hypnea charoides‐valentiae complex (Rhodophyta, Gigartinales) in Japan is investigated on the basis of field‐collected and cultured plants. We can recognize two species, Hypnea charoides Lamouroux and H. flexicaulis Yamagishi et Masuda, sp. nov., which are distinguished by: (i) conspicuous percurrent main axes growing straight in the former versus less conspicuous percurrent main axes growing flexuosely in the latter; (ii) narrow branching angles (less than 90°) in the former versus wide branching angles (up to 150°) in the latter; (iii) the presence of branches and branchlets showing abrupt adaxial bending in the former versus those showing pronounced abaxial bending, some growing into hooked branchlets in the latter; and (iv) the absence (in the former) or presence (in the latter) of antler‐like branches. Nucleotide sequences of the plastid‐encoded large subunit of the ribulose‐1,5‐bisphosphate carboxylase/oxygenase gene (rbcL) support the differentiation of these two entities: the pairwise distance between them was 83 b.p. (6.2%).     

A new potentially toxic Azadinium species (Dinophyceae) from the Mediterranean Sea,A. dexteroporum sp. nov.

A new photosynthetic planktonic marine dinoflagellate, Azadinium dexteroporum sp. nov., is described from the Gulf of Naples (South Tyrrhenian Sea, Mediterranean Sea). The plate formula of the species, Po, cp, X, 4′, 3a, 6″, 6C, 5?S, 6? and 2″″, is typical for this recently described genus. Azadinium dexteroporum is the smallest rep‐resentative of the genus (8.5 μm average length, 6.2 μm average width) and shares the presence of a small antapical spine with the type species A. spinosum and with A. polongum. However, it differs from all other Azadinium species for the markedly asymmetrical Po plate and the position of the ventral pore, which is located at the right posterior end of the Po plate. Another peculiarity of A. dexteroporum is the pronounced concavity of the second intercalary plate (2a), which appears collapsed with respect to the other plates. Phylogenetic analyses based on the large subunit 28S rDNA (D1/D2) and the internal transcribed spacer (ITS rDNA) support the attribution of A. dexteroporum to the genus Azadinium and its separation from the other known species. LC/MS‐TOF analysis shows that Azadinium dex‐teroporum produces azaspiracids in low amounts. Some of them have the same molecular weight as known compounds such as azaspiracid‐3 and ‐7 and Compound 3 from Amphidoma languida, as well as similar fragmentation patterns in some cases. This is the first finding of a species producing azapiracids in the Mediterranean Sea.     

Molecular systematics and Holarctic phylogeography of cestodes of the genus Anoplocephaloides Baer, 1923 s. s. (Cyclophyllidea,Anoplocephalidae) in lemmings (Lemmus,Synaptomys)

The present molecular systematic and phylogeographic analysis is based on sequences of cytochrome c oxidase subunit 1 (cox1) (mtDNA) and 28S ribosomal DNA and includes 59 isolates of cestodes of the genus Anoplocephaloides Baer, 1923 s. s. (Cyclophyllidea, Anoplocephalidae) from arvicoline rodents (lemmings and voles) in the Holarctic region. The emphasis is on Anoplocephaloides lemmi (Rausch 1952) parasitizing Lemmus trimucronatus and Lemmus sibiricus in the northern parts of North America and Arctic coast of Siberia, and Anoplocephaloides kontrimavichusi (Rausch 1976) parasitizing Synaptomys borealis in Alaska and British Columbia. The cox1 data, 28S data and their concatenated data all suggest that A. lemmi and A. kontrimavichusi are both non‐monophyletic, each consisting of two separate, well‐defined clades, that is independent species. As an example, the sister group of the clade 1 of A. lemmi, evidently representing the ‘type clade’ of this species, is the clade 1 of A. kontrimavichusi. For A. kontrimavichusi, it is not known which one is the type clade. There is also fairly strong evidence for the non‐monophyly of Anoplocephaloides dentata (Galli‐Valerio, 1905)‐like species, although an earlier phylogeny suggested that this multispecies assemblage may be monophyletic. The results suggest a deep phylogenetic codivergence of Lemmus spp. and A. lemmi, primarily separating the two largely allopatric host and parasite species at the Kolyma River in east Siberia. There are also two allopatric sublineages within each main clade/species of A. lemmi and Lemmus, but the present distributions of the sublineages within the eastern L. trimucronatus and clade 1 of A. lemmi are not concordant. This discrepancy may be most parsimoniously explained by an extensive westward distributional shift of the easternmost parasite subclade. The results further suggest that the clade 1 of A. kontrimavichusi has diverged through a host shift from the precursor of L. trimucronatus to S. borealis.     

Identification and field and laboratory tests of the sex pheromone of Cerconota anonella Sepp. (Lepidoptera: Oecophoridae)

The Annona fruit borer, Cerconota anonella Sepp., is a serious agricultural pest in many tropical areas of the world. The identification of an attractant for male C. anonella could offer new methods for pest detection and control. A mixture of compounds extracted from female C. anonella elicited antennal depolarization in the male borer. These compounds were identified as octadecanal, 1‐octadecanol, octadecyl acetate, (Z)‐octadec‐9‐enal (Z9‐18:Ald), (Z)‐octadec‐9‐en‐1‐ol (Z9‐18:OH) and [(Z)‐octadec‐9‐enyl] acetate (Z9‐18:Ac) by one‐ and two‐dimensional gas chromatography–mass spectrometry. In laboratory bioassays, synthetic individual compounds as well as synthetic mixtures were found to be attractive to males. In addition, field tests using Delta traps with 1 mg of the ternary mixture composed of Z9‐18:Ac, Z9‐18:Ald and Z9‐18:OH in the ratio of 1 : 3 : 5 caught as many males as traps containing virgin females. The ternary mixture of Z9‐18:Ac, Z9‐18:Ald and Z9‐18:OH was identified as attractant to C. anonella males and can be used to detect and control populations of this insect in Annonaceae plantations.     

Evolution of MHC-<Emphasis Type="Italic">DRB</Emphasis> class II polymorphism in the genus <Emphasis Type="Italic">Apodemus</Emphasis> and a comparison of <Emphasis Type="Italic">DRB</Emphasis> sequences within the family Muridae (Mammalia: Rodentia)

Allelic diversity at major histocompatibility complex (MHC) genes is thought to be maintained by balancing selection over long periods of time, even across multiple speciation events. Trans-species sharing of MHC alleles among genera has been supported by many studies on mammals and fish, but in rodents, the results are ambiguous. We investigated natural levels of MHC-DRB variability and evolutionary processes in the wood mouse (Apodemus sylvaticus) and the yellow-necked mouse (Apodemus flavicollis), which are common, sympatric murid rodents in European forests. Using single-strand conformation polymorphism analysis and DNA sequencing, 38 DRB exon 2 alleles were detected among 162 A. sylvaticus from nine different locations in Germany and Switzerland, and 15 DRB exon 2 alleles were detected among 60 A. flavicollis from three different locations in northern Germany. There was evidence for balancing selection in both species. Phylogenetic analysis, including additional murid taxa, showed that the DRB exon 2 sequences did not separate according to species, consistent with trans-species evolution of the MHC in these taxa.     

A morphological and molecular assessment of the genus Prionitis J. Agardh (Halymeniaceae, Rhodophyta)

A critical reassessment of the morphological features of two closely related red algal genera, Grateloupia C. Agardh and Prionitis J. Agardh (Halymeniaceae), shows that members of the two genera share very similar reproductive (including the Grateloupia‐type auxiliary‐cell ampullae) and vegetative characters. Diagnostic features hitherto used for distinguishing these two genera, the texture of blades (lubricous to leathery in Grateloupia vs cartilaginous in Prionitis) and the position of reproductive structures (scattered over the entire blade in Grateloupia vs confined to particular portions of the blade in Prionitis), are continuous across some 75 species of both genera, thus making it difficult to draw a clear‐cut distinction between the two genera. In ribulose‐1,5‐bisphosphate carboxylase/oxygenase gene (rbcL) sequence analyses, the species of Grateloupia and Prionitis, including the two generitypes, constitute a large monophyletic clade in the Halymeniaceae. It is therefore proposed that Prionitis be included in the synonymy under Grateloupia and the appropriate combinations are proposed.     

Comparative functional morphology of mandibular forward movement during mastication of two murid rodents,Apodemus speciosus (Murinae) and Clethrionomys rufocanus (Arvicolinae)

The anatomy of the masticatory apparatus, the direction in which masticatory muscles act during mastication, and jaw muscle forces as estimated by muscle dry weight are compared between two murid rodents, the Japanese field mouse (Apodemus speciosus; subfamily Murinae) and the gray red-backed vole (Clethrionomys rufocanus; subfamily Arvicolinae). The occlusal forces exerted by the deep masseter and the anterior temporalis are large in C. rufocanus. Furthermore, in this species, the angle between the sagittal plane and the occlusal plane of the cheek teeth is larger than in A. speciosus. Therefore, a relatively large occlusal force can be generated in C. rufocanus. The estimated line of action of the anterior temporalis differs markedly between these two species. The functional significance of this difference is discussed relative to the adaptive dental characteristics for food processing, the forces required to masticate different types of food, and the forces that control mandibular forward movement. J Morphol 231:131–141, 1997. © 1997 Wiley-Liss, Inc.