Nest predation is the main cause of hatching failure for many turtle populations. For green turtles (Chelonia mydas) nesting at Chagar Hutang in Redang Island, Malaysia, Asian water monitors (Varanus salvator) are a potential nest predator. However, no studies have documented the space use of this species in coastal habitat adjacent to a sea turtle nesting beach to assess its potential impact on turtle nests. Here, we used Global Positioning System (GPS) data loggers to quantify space use of Asian water monitors in order to establish the extent to which they use sea turtle nesting areas. Asian water monitors had a diurnal activity pattern and spent most of their time in rain forest habitat behind the sea turtle nesting beach. The home range occupied by Asian water monitors varied between 0.015 and 0.198 km² calculated by the Kernel Brownian Bridge method. The space use patterns of individual Asian water monitors varied between individuals. Two males had relatively small home ranges, whereas one male and the female had a relatively large home range. Because tracked Asian water monitors in this study rarely visited the sea turtle nesting areas, it is probable that only a few individuals are responsible for opening nests.

Foam nests of frogs are natural biosurfactants that contain potential compounds for biocompatible materials, Drug Delivery System (DDS), emulsifiers, and bioremediation. To elucidate the protein components in the foam nests of Rhacophorus arboreus, which is an endemic Japanese frog species commonly seen during the rainy season, we performed amino acid analysis, SDS-PAGE electrophoresis, and matrix-assisted laser desorption/ionization mass spectrometry using intact foam nests. Many proteins were detected in these foam nests, ranging from a few to several hundred kDa, with both essential and non-essential amino acids. Next, we performed transcriptome analysis using a next-generation sequencer on total RNAs extracted from oviducts before egg-laying. The soluble foam nests were purified by LC-MS and analyzed using Edman degradation, and the identified N-terminal sequences were matched to the transcriptome data. Four proteins that shared significant sequence homologies with extracellular superoxide dismutase of Nanorana parkeri, vitelline membrane outer layer protein 1 homolog of Xenopus tropicalis, ranasmurfin of Polypedates leucomystax, and alpha-1-antichymotrypsin of Sorex araneus were identified. Prior to purification of the foam nests, they were treated with both a reducing reagent and an alkylating agent, and LC-MS/ MS analyses were performed. We identified 22 proteins in the foam nests that were homologous with proteinase inhibitors, ribonuclease, glycoproteins, antimicrobial protein and barrier, immunoglobulin-binding proteins, glycoprotein binding protein, colored protein, and keratin-associated protein. The presence of these proteins in foam nests, along with small molecules, such as carbohydrates and sugars, would protect them against microbial and parasitic attack, oxidative stress, and a shortage of moisture.

Oviparous, ovoviviparous and viviparous reproduction are interesting subjects for understanding animals' evolutionary pathways and adaptation to their life history and habitat conditions. In this study, we examined the reproductive mode of the ovoviviparous mayfly Cloeon dipterum, particularly comparing embryogenesis between hand-pairing and unmated females' common oviduct. Our study suggested that the high developmental rate of C. dipterum observed in a recent study could be ascribed to their absorption of unfertilized eggs. The developmental rates of hand-paired females were almost 100%, while their egg-bearing numbers were lower than those of virgin females. Thus, such reduced egg numbers suggest the maternal absorption of unfertilized eggs. This trait is thought to have evolved with the ovoviviparous characteristics of C. dipterum. We identified the basis of the irregularity of this species exhibiting such a high (i.e., 100%) developmental rate in our previous recent study.

The spatiotemporal expression of zygotic genes is regulated by transcription factors, which mediate cell fate decision and morphogenesis. Investigation of the expression patterns and their transcriptional regulatory relationships is crucial to understand embryonic development. Staged RNA-seq of the ascidian Halocynthia roretzi has previously shown that nine genes encoding transcription factors are transiently expressed at the blastula stage, which is the stage at which cell fates are specified and differentiation starts. Six of these transcription factors have already been found to play important roles during early development. However, the functions of the other transcription factors (FoxJ-r, SoxF, and SP8/9) remain unknown. The study of the spatial and temporal expression patterns showed that all three genes were expressed in the animal hemisphere as early as the 16-cell stage. This is likely due to transcription factor genes that are expressed in the vegetal hemisphere, which have been extensively and comprehensively analyzed in previous studies of ascidians. Functional analyses using FoxJ-r morphants showed that they resulted in the disruption of laterality and the absence of epidermal mono-cilia, suggesting FoxJ-r functions in cilia formation and, consequently, in the generation of left-right asymmetry, as observed in vertebrates. SoxF knockdown resulted in incomplete epiboly by the ectoderm during gastrulation, while SP8/9 knockdown showed no phenotype until the tailbud stage in the present study, although it was expressed during blastula stages. Our results indicate that transcription factor genes expressed at the cleavage stages play roles in diverse functions, and are not limited to cell fate specification.

Southern Africa has a diverse endemic scorpion fauna, but a paucity of information currently confounds conservation of the group. Phylogeographic approaches represent a useful tool to identify the patterns and processes which underpin scorpion diversity, but these studies are lacking for southern African species. Among southern African scorpions, the semi-lithophilous Opistophthalmus pallipes has strict habitat requirements, and a distribution historically subjected to profound environmental turnover. As such, the species offers a model system to investigate the interplay between intrinsic and extrinsic factors as drivers of diversity and endemism. To investigate spatial genetic patterns within O. pallipes and the possible drivers thereof, the current study combines mitochondrial DNA and ecological information under a phylogeographic approach. The species is characterized by several genetically discrete and divergent populations. The factors which shape these genetic patterns appear to be both intrinsically (ecological specificity) and extrinsically (landscape structure and ecogeographic conditions) influenced, with major divergences corresponding to periods of profound environmental changes. Taken together, the findings of this study provide evidence of spatial genetic isolation and genetic diversity within a stenotopic southern African scorpion species. These findings partly explain the staggering diversity and endemism in southern African scorpions, but further phylogeographic studies are necessary to propose conservation scenarios for this group.

In medaka fishes, the family Adrianichthyidae, tropical species are known to be sexually more dimorphic than temperate species. If this reflects that tropical species are exposed to stronger sexual-selection pressures than temperate species, mating behaviors may also differ between tropical and temperate species. Our mating experiments revealed that males of Oryzias woworae, a tropical species of the family, perform “chasing” another male more frequently than males of O. sakaizumii, a temperate congener, and that male–male “combats” of O. woworae tended to be followed by chasing compared with combats of O. sakaizumii males, indicating that O. woworae males are more aggressive in male–male interactions than O. sakaizumii males. Males of O. woworae also performed “approaching” a female and “mating dance” more frequently than O. sakaizumii males, indicating that O. woworae males are also more active in courting females. Males of O. sakaizumii often omitted “mating dance” in their mating sequences, supporting this view. Moreover, O. woworae females tended to reject male “wrapping”, an attempt for fertilization, more frequently than O. sakaizumii females, suggesting that O. woworae females are choosier in mating than O. sakaizumii females. These findings are concordant with the view that O. woworae is exposed to stronger sexual-selection pressures than O. sakaizumii.

In vertebrates, gonadotropin-releasing hormone (GnRH) regulates gonadal maturation by stimulating the synthesis and release of pituitary gonadotropins. GnRH has also been identified in invertebrates. Crustacea consists of several classes including Cephalocarida, Remipedia, Branchiopoda (e.g., tadpole shrimp), Hexanauplia (e.g., barnacle) and Malacostraca (e.g., shrimp, crab). In the malacostracan crustaceans, the presence of GnRH has been detected in several species, mainly by immunohistochemistry. In the present study, we examined whether a GnRH-like peptide exists in the brain and/or nerve ganglion of three classes of crustaceans, the tadpole shrimp Triops longicaudatus (Branchiopoda), the barnacle Balanus crenatus (Hexanauplia), and the hermit crab Pagurus filholi (Malacostraca), by immunohistochemistry using a rabbit polyclonal antibody raised against chicken GnRH-II (GnRH2). This antibody was found to recognize the giant freshwater prawn Macrobrachium rosenbergii GnRH (MroGnRH). In the tadpole shrimp, GnRH-like-immunoreactive (ir) cell bodies were located in the circumesophageal connective of the deuterocerebrum, and GnRH-like-ir fibers were detected also in the ventral nerve cord. In the barnacle, GnRH-like-ir cell bodies and fibers were located in the supraesophageal ganglion (brain), the subesophageal ganglion, and the circumesophageal connective. In the hermit crab, GnRH-like-ir cell bodies were detected in the anterior-most part of the supraesophageal ganglion and the subesophageal ganglion. GnRH-like-ir fibers were observed also in the thoracic ganglion and the eyestalk. These results suggest that a GnRH-like peptide exists widely in crustacean species.

The taxonomic status of extinct Japanese or Honshu wolves (Canis lupus hodophilax) has been disputed since the name hodophilax was first proposed by Temminck in 1839 on the basis of specimens stored in Leiden, the Netherlands. Points of controversy include whether the type specimen of hodophilax (Jentink c: RMNH.MAM.39181) and the other two specimens from Leiden (Jentink a: RMNH.MAM.39182 and Jentink b: RMNH.MAM.39183) represent different varieties or subspecies of Japanese wolves or not. Two Japanese names, ookami and jamainu, used to describe wild Canis species, further complicate the issue. In this study, the taxonomic status of Japanese wolves was clarified using mitochondrial DNA of the three specimens stored at the Naturalis Biodiversity Center in Leiden, in addition to three Japanese wolf specimens stored at the Museum für Naturkunde in Berlin and five new samples from Japan. The mitochondrial genomes of the type specimen of hodophilax (Jentink c) and another sample from Leiden (Jentink b) as well as Berlin specimens were included in the cluster of Japanese wolves distinct from other grey wolves. However, the other sample from Leiden (Jentink a) was identified as a domestic dog. A mitochondrial genome analysis suggested that Japanese wolves could be categorized into two distinct clusters. Studies of nuclear genomes are needed to further clarify the taxonomic status, divergence time, and population genetic structure of Japanese wolves.

Almost all viviparous species possess male external genitalia; for example, the mammalian penis is an intromittent organ. Some live-bearing bony fish use their anal fins to assist in mating and internal fertilization. We previously reported a male-specific asymmetric curvature at the posterior end of the anal fin in Xenotoca eiseni, a viviparous fish of the family Goodeidae. However, three other goodeid species, Xenotoca melanosoma, Zoogoneticus quitzeoensis, and Chapalichthys pardalis, examined in that study possessed lesser anal fin curvature modifications as compared to those in the anal fin of X. eiseni. Here, we report the second case of acute-angled curvature modification of the male anal fin in the family Goodeidae. We obtained a dead specimen of the goodeid species Xenotoca variata from a city zoo in Japan, and the morphological and histological analyses indicated an acute-angled asymmetric curvature of the posterior end of the anal fin in X. variata, similar to that observed in X. eiseni in the previous study. However, in our previous report, obtuse-angled modification was only observed in one other Xenotoca species, X. melanosoma, and two species belonging to other genera, Z. quitzeoensis and C. pardalis. Therefore, our findings suggest that the acute-angled curvature in the male anal fin has been developed in the genus Xenotoca.

Eastern broad-toothed field mouse, Apodemus mystacinus, is a rocky habitat dwelling rodent distributed in Asia Minor, the Levant, the Caucasus, and the Zagros Mountains. In this study, we investigated the phylogenetic relationship between different populations of A. mystacinus throughout its range, based on the mitochondrial cytb marker. Phylogenetic analyses revealed the existence of five separately evolving lineages within A. mystacinus, of which two previously unrecognized lineages were identified in the Zagros Mountains and the Levant. Divergence between two major clades of the subgenus Karstomys, corresponding to A. mystacinus and Apodemus epimelas, is inferred to coincide with the Messinian Salinity Crisis (Late Miocene), whereas the splits between major lineages of A. mystacinus are inferred to have occurred during the Pleistocene. Colonization of the Zagros may have occurred from different refugia via eastward migration of the Turkish population and then again by a more recent colonization from the Caucasus, after reopening of the land corridor between the Caucasus and the Zagros Mountains during the Holocene drought.

Many insects show daily and circadian changes in morphology and physiology in their compound eye. In this study, we investigated whether the compound eye had an intrinsic circadian rhythm in the cricket Gryllus bimaculatus. We found that clock genes period (per), timeless (tim), cryptochrome 2 (cry2), and cycle (cyc) were rhythmically expressed in the compound eye under 12-h light/12-h dark cycles (LD 12:12) and constant darkness (DD) at a constant temperature. After the optic nerves were severed (ONX), a weak but significant rhythmic expression persisted for per and tim under LD 12:12, while under DD, tim and cyc showed rhythmic expression. We also found that more than half of the ONX compound eyes exhibited weak but significant circadian electroretinographic rhythms. These results clearly demonstrate that the cricket compound eye possesses an intrinsic circadian oscillator which can drive the circadian light sensitivity rhythm in the eye, and that the circadian clock in the optic lobe exerts its influence on the oscillator in the eye.

Two new species of the cockroach genus EucorydiaHebard, 1929 from the Nansei Islands in Southwest Japan were compared to two closely related congeners, Eucorydia yasumatsuiAsahina, 1971 and Eucorydia dasytoides (Walker, 1868). Eucorydia donanensis Yanagisawa, Sakamaki, and Shimano sp. nov. from Yonaguni-jima Island was characterized by an overall length of 12.5–14.5 mm in males. The dorsal side of the male abdomen was entirely dark purple and there was an obscure orange band running down the middle of the tegmen. Eucorydia tokaraensis Yanagisawa, Sakamaki, and Shimano sp. nov. was characterized by an overall length of 12.0–13.0 mm in males and a distinct orange band running down the middle of the tegmen. Eucorydia yasumatsui, E. donanensis, E. tokaraensis and the zonata population of E. dasytoides were divided into four lineages in a maximum-likelihood tree generated from a dataset concatenated from five (two nuclear, 28S rRNA, histone H3, and three mitochondrial, COII, 12S rRNA, 16S rRNA) genes. We recognized the three Japanese lineages E. yasumatsui, E. donanensis, and E. tokaraensis as distinct species, which were also supported by the pairwise genetic distances (5.4–7.8%, K2P) of the COI sequences. Morphometric analysis was performed on the genitalia. A principal component analysis plot revealed that the sizes of the genitalia in the three Japanese species were similar to each other and smaller than that of the zonata population of E. dasytoides. The analysis also revealed that the three Japanese species were distinguished from each other by combinations of the sizes of L3 and L7 sclerites and the shape of R2 sclerite, with some overlapping exceptions.