The mechanisms for the coexistence of multiple species occupying the same ecological niche are often puzzling. Predator effects on competitively superior species is one possible mechanism. In this study, we tested whether the presence of size-selective predators (fishes) acts as a mechanism for the coexistence of two species of case-bearing caddisfly larvae, Perissoneura paradoxa and Psilotreta kisoensis (Odontoceridae, Trichoptera). The larvae of these two species have similar ecological and life history traits except their body size, and they have been found to coexist only in habitats shared with predatory fishes. Experiments on intra and interspecific competition revealed that the larger Pe. paradoxa always outcompeted the smaller Ps. kisoensis in the absence of predatory fishes, suggesting that Pe. paradoxa performed intra-guild predation on Ps. kisoensis. We also conducted experiments to examine how strongly each of these species responded in terms of case repair with/without a predator chemical cue after their cases were partly dismantled. Perissoneura paradoxa exhibited a stronger case repair response in the presence of a predator chemical cue than that exhibited by Ps. kisoensis, suggesting that Pe. paradoxa is more vulnerable to fish predation, probably because their body size is in the preferred prey range of fishes. We suggest that the presence of predators works in the favor of smaller, subordinate species through size-selective predator effects, enabling these two competitive species to coexist in the same habitat.

It is important to investigate visual orienting in reptiles to better understand the basic organization of the oculomotor system in vertebrates. However, quantitative analyses of visual orienting behavior in reptiles have rarely been conducted, except in chameleons. In the present study, we videorecorded the head and body movements of the lizard Takydromus tachydromoides during visual tracking of moving prey and analyzed them frame-by-frame. Before approaching prey, visual tracking mainly consisted of brief intermittent turns of the head (saccade). After the head saccades, the angular position of the prey relative to the lizard head was kept at 10–70° (laterally) in most cases, rather than at 0° (in front). In addition, the ratio of the amplitude of the head saccades to prey position was 0.2–0.3, which is much smaller than 1, suggesting that the head did not orient exactly toward the prey after most saccades. These results were observed under both white (homogeneous) and grating (structured) backgrounds. Possible functions of head saccades in the lizard are discussed.

Embryonic mouse submandibular epithelia initiate branching morphogenesis within two days when embedded in Matrigel and stimulated by members of the epidermal growth factor family. However, it is unknown whether the end buds further branch over longer culture periods, and whether saliva-producing cells differentiate there. In the present study, we cultivated three major (submandibular, sublingual and parotid) salivary epithelia from 13-day embryos for 14 days in mesenchyme-free cultures. All epithelia continued to grow and branch to form numerous acinus-like structures in medium supplemented with neuregulin 1, fibroblast growth factor 1, and insulintransferrin-sodium selenite. Alcian blue staining to detect mucous cells showed that each epithelium differentiated via three distinct modes, as seen in normal development, although the staining intensities were weaker than in normal development. RT-PCR analysis of the amylase gene showed that no epithelia expressed amylase after 14 days of culture, which is inconsistent with the fact that only parotid epithelium does so at postnatal day 7 during normal development. These results suggest that cytodifferentiation progresses to a lesser extent in mesenchyme-free cultures than in vivo.

The major histocompatibility complex (MHC) includes many genes that are essential for the adaptive immune system, and variation in the antigen binding site (ABS) is related to resistance against pathogens. In the present study, quantitative real-time PCR indicated a larger number of MHC gene copies in the endangered population of Blakiston's fish owl (Bubo blakistoni) than in five other owl species, and massively parallel pyrosequencing detected more MHC class IIβ per individual alleles in B. blakistoni than in the other species. A chromosomal fluorescence in situ hybridization (FISH) analysis showed that the MHC class I and class IIβ loci are closely linked on a single pair of microchromosomes, indicating that the MHC genes were tandemly duplicated in a limited chromosomal region. Because B. blakistoni has twice as many MHC genes as its sister species, the tawny fish owl (Bubo flavipes), the duplication of MHC genes occurred after these species diverged by speciation. A Bayesian molecular phylogenetic analysis showed that the DAB1 and DAB2 lineages of MHC class IIβ alleles from various strigid species each formed a separate clade, indicating that the two allelic lineages preceded the radiation of Strigidae and evolved as paralogs. By contrast, the ABS sequences did not form distinct clades between DAB1 and DAB2 alleles but were intermixed, presumably due to gene conversion. Despite the low diversity of alleles per locus, B. blakistoni had many lineages of MHC class IIβ alleles. Gene duplication increases variation in the MHC genes in this species, and could have facilitated adaptation in small populations.

Regions of Iran, Afghanistan, Pakistan and northwestern India have been proposed as the place of origin of Mus musculus castaneus. But despite the fact that Pakistan encompasses an important part of its range, M. m. castaneus populations in Pakistan have not been the subject of intensive genetic and biogeographic studies, except for a very small number of samples included in past studies. We studied genetic variation in M. m. castaneus (CAS) from northern Punjab Province, Pakistan, by using cytochrome b (Cytb) analysis in a sample of 98 individuals. Median-joining network revealed four well differentiated CAS sub-lineages coexisting within a small geographical region; these had previously been thought to have largely non-overlapping geographic distributions. Moreover, haplotypes from Pakistan occupied a central position in the network and all identified global haplotypes were also present in Pakistan. All identified CAS sub-lineages proved to be highly diverse on the basis of haplotype and nucleotide diversity indices. Tajima's D test and Fu's Fs tests of neutrality suggest recent population expansions in all sub-lineages. Expansion times were estimated as 21,760–134,930, 10,800–64,400 and 4950–30,665 ybp using substitution rates of 2.5%, 5% and 11%, respectively. Our results support the hypothesis that northern Punjab Province in Pakistan is the most likely source area for M. m. castaneus, and that the CAS sub-lineages in this region have undergone rapid population expansion events at different time periods, which appear to have benefitted from human-mediated transport, although one of them clearly predates the establishment of human settlements in this region.

The Alpheus brevicristatus De Haan, 1844 is one of the commonest shrimp species inhabiting the tidal flats in Japan. This species is sometimes accompanied by the facultative symbiotic goby, Acentrogobius spp. Here, we investigated the burrow morphology of A. brevicristatus in a tidal flat of Uranouchi Inlet, Kochi Prefecture, Japan. We also reviewed existing literature on alpheid burrow morphology using the resin casting technique, to determine how burrows vary in the presence and absence of gobies. Nine burrows were casted in situ using polyester resin. All burrows were long, but shallow in structure, with several funnel-shaped openings and short cul-de-sac branches. This species appears to use several burrow openings to access the sediment surface for feeding with high efficiency. Gobies were not associated with all burrows cast; however, 1–3 individuals of the small alpheid shrimp Athanas japonicus Kubo, 1936 were entombed in seven of the casts. A review of 12 studies on the burrow morphology of 16 Alpheus species based on resin casting techniques showed wide variation in burrow characteristics, such as burrow depth, length, and number of openings. Our findings suggest that burrow structure is influenced by species-specific characteristics and sediment type. The possibility that the presence of the symbiotic goby affects the burrow morphology of Alpheus shrimp is discussed.

In recent decades, variation in female preferences within a species has attracted considerable attention. However, how female individuals in most species change their mate preferences remain unclear. In this study, we examined the change in mate preferences in female individuals of the guppy, Poecilia reticulata. This was done by exposing naïve, virgin females to two male groups differing in the abundance of ornamented males over a period of seven days. The male-exposure treatment groups consisted of (1) primarily males with large orange spots and (2) primarily males with small orange spots. Following each treatment, we measured female preference for brightly colored males with large orange spots, using a pair of digitally modified male images differing in orange spot patterns. Our results indicated that the patterns of change in mate preferences differed between young and old females. After exposure to primarily males with large orange spots, young females were more attracted to the male images overall and showed a preference for the bright male image. Specifically, young females showed a stronger preference for the bright male image when they had been exposed to male groups in which the among-male difference in orange spot patterns was greater. However, in old females, both male-exposure treatments and order of the treatments affected their mate preferences. These results suggest that social components, i.e., both the abundance of ornamented males and the age structure of females in a population, influence the direction and intensity of sexual selection.

The occurrence of black fur, or melanism, in many mammalian species is known to be linked to DNA sequence variation in the agouti signaling protein (Asip) gene, which is a major determinant of eumelanin and pheomelanin pigments in coat color. We investigated 38 agouti (i.e., banded wildtype) and four melanistic Rattus rattus species complex (RrC) lineage II specimens from Okinawa Island, Ryukyu Islands, Japan, for genetic variation in three exons and associated flanking regions in the Asip gene. On Okinawa, a predicted loss-of-function mutation caused by a cysteine to serine amino acid change at p.124C>S (c.370T>A) in the highly conserved functional domain of Asip was found in melanistic rats, but was absent in agouti specimens, suggesting that the p.124C>S mutation is responsible for the observed melanism. Phylogeographic analysis found that Asip sequences from Okinawan RrC lineage II, including both agouti and melanistic specimens, differed from: 1) both agouti and melanistic RrC lineage I from Otaru, Hokkaido, Japan, and 2) agouti RrC lineages I and II from South Australia. This suggests the possibility of in-situ mutation of the Asip gene, either within the RrC lineage II population on Okinawa or in an unsampled RrC lineage II population with biogeographic links to Okinawa, although incomplete lineage sorting could not be ruled out.

Antimicrobial peptides (AMPs) were previously isolated from the skin of the Ryukyu brown frog Rana okinavana. However, this species has recently been reclassified as two species, i.e., Rana kobai and Rana ulma. As a result, it was determined that AMPs isolated from R. okinavana were in fact products of R. kobai, but not of R. ulma. In the present study, we collected skin samples from the species R. ulma and cloned twelve cDNAs encoding AMP precursors for the acyclic brevinin-1ULa--1ULf, the temporin-ULa-ULc, ranatuerin-2ULa, japonicin-1ULa, and a novel peptide using reverse-transcription polymerase chain reaction techniques. The deduced amino acid sequence of the novel peptide had a high similarity to those of Rana chensinensis chensinin-1CEa--1CEc, which were cloned by Zhao et al. (2011), but had a low similarity with R. chensinensis chensinin-1, which was cloned by Shang et al. (2009). To avoid confusion with these two different chensinin-1 families, we termed our peptide ulmin-1. Among these peptides, we focused on two peptides, brevinin-1ULf and ulmin-1ULa, and examined the antimicrobial and cytotoxic activity of their synthetic replicates. In broth microdilution assays, growth inhibitory activities against Staphylococcus aureus, Bacillus cereus, and Candida albicans were detected for brevinin-1ULf but not for ulmin-1ULa, whereas scanning electron microscopic observations revealed that both peptides induce morphological abnormalities in these microbes. In addition, binding activity of ulmin-1ULa to the bacterial cell wall component lipoteichoic acid was higher than that of brevinin-1ULf. In contrast, hemolytic and cytotoxic activities of brevinin-1ULf were stronger than those of ulmin-1ULa.

The terrestrial slug Limax has been used as a model animal for studying the neural mechanisms underlying associative olfactory learning. The slug also innately exhibits negative phototactic behavior using its eyes. In the present study, we developed an experimental paradigm for quantification of slug's negative phototaxis behavior, and investigated whether the nature of the negative phototaxis can be modified by learning experience. The experimental set-up consists of light and dark compartments, between which the slug can move freely. During conditioning, the slug was placed in the light compartment, and an aversive stimulus (quinidine sulfate solution) was applied when it reached the dark compartment. After a single conditioning session, the time to reach the dark compartment significantly increased when it was tested following 24 hr or one week. Protein synthesis inhibition immediately following the conditioning impaired the memory retention at one week but not at 24 hr. The retrieval of the memory was context-dependent, as the time to reach the dark compartment did not significantly increase if the slug was placed on a floor with a different texture in the memory retention test. If the aversive stimulus was applied when the slug was in the light compartment, the time to reach the dark compartment did not increase after 24 hr. This is the first report demonstrating the capability of the slug to form context-dependent passive avoidance memory that can be established in a single conditioning session.

Examination of the holotype of Hynobius stejnegeri Dunn, 1923 revealed that the species is identical with H. yatsui Oyama, 1947, recently resurrected from H. naevius (Temminck and Schlegel, 1838). The name H. yatsui is therefore relegated to a subjective junior synonym of H. stejnegeri. The latter name, however, has been widely applied to an amber-colored or tortoise-shell salamander from Kyushu, southwestern Japan, which now requires a new name. We, thus, describe the ambercolored salamander as a new species Hynobius ikioi. The new species is phylogenetically very close to H. amakusaensis Nishikawa and Matsui, 2014 and H. osumiensis Nishikawa and Matsui, 2014, both also from Kyushu, but is easily distinguished from them by its uniquely bi-colored dorsum.