Through population expansion and accidental or deliberate introduction, prey commonly encounter novel predators they had never seen before. Several studies have shown that animals can generalize their learned recognition of a familiar predator to novel ones according to predators' identical or similar features. This process in fish mainly depends on the visual and chemosensory cues they receive. However, there is a lack of understanding of the different effects of these two cues. Topmouth gudgeons (Pseudorasbora parva) that had never seen turtles were captured and used as the subjects, and three freshwater turtles of different genera were used as predators. Before and after using one turtle for predator training treatment of topmouth gudgeons, fish responses to visual and chemosensory cues of each turtle were tested and recorded, and it was found that predator training promoted topmouth gudgeons' recognition of the risks represented by visual cues of all three turtles and by chemosensory cues of the turtle that were used in training. These results further verify the generalization of predator recognition in fish and indicate that visual cues have a more extensive effect on fish than chemosensory cues in identifying novel predators, especially predators that are distantly related to the familiar threats.

A mucous secreting organ in ascidians, the endostyle, consists of several epithelial zones with different ciliary length, density, and beating direction. Here we found by transmission electron microscopy that long cilia in endostyle zone 1 showed 9 + 2 axonemal structures but completely lacked the outer arm dynein. In contrast, cilia in other zones bore both outer and inner dynein arms. Western blotting and immunofluorescence microscopy further revealed that zone 1 appeared to lack not only outer arm dynein but also two-headed inner arm dynein. These results suggest a mechanism for a region-specific gene suppression that causes the limited loss of two-headed axonemal dyneins in the endostyle epithelium. The loss of these dyneins in zone 1 is considered to contribute to the generation of undulating ciliary movement that is essential for a unique circuit of mucus flow in the endostyle.

Perinereis nuntia is a fully segmented worm with complete intersegmental septa. A previous study of females revealed that germ cells of this animal originate in the tail end segment, called the pygidium. Germ cells were duplicated in the pygidium, transferred to a newly generated segment, and then settled in the parapodia. Within each segment, the settled germ cells proliferated in the parapodia and then migrated into a body cavity area to begin meiotic development. Currently, there is not much information about differences between male and female germ cell development. Therefore, we conducted monthly in situ hybridization analyses using the germ cell marker Pn-piwi and histological examinations. Germ cells detected by Pn-piwi initially settled in the distal areas of the parapodia on both sides of each segment, then formed a large germ cell cluster in each parapodium, and finally, small germ cell clusters were formed by the separation of the large clusters. The small clusters migrated to the deeper body cavity area during growth by segment addition. Until the female germ cells began vitellogenesis, the sex of germ cells could not be identified by morphological observation. Thus, male and female P. nuntia may have the same mechanism of germ cell provision to all segments. At the time of spawning, sperm were released from nephridiopores at the 2^nd through 15^th segments from the pygidium, while eggs were released through ruptures in the skin of 2–3 segments between the 10^th and 30^th segments from the tail.

We surveyed the genetic structure of Hynobius nigrescens Stejneger, 1907, a lentic breeding salamander widespread throughout montane and lowland regions of northeastern Japan. We performed a mitochondrial DNA analysis to explore intraspecific genetic variation and infer the evolutionary population history of H. nigrescens. Complete 1141 bp sequences of the mitochondrial cytochrome b gene were studied for 134 adult and larval individuals collected from 62 localities, encompassing the known range of the species. Hynobius nigrescens proved to be monophyletic, including two major clades (Clade II from southwestern Hokuriku and Clade I from all other localities). The latter clade comprises four well-supported and geographically structured subclades, which show genetic distances smaller than those seen in the widely sympatric species Hynobius lichenatus. Results of population statistical analyses indicated that Clade II of the westernmost range of H. nigrescens seems to have maintained a constant population size, while Clade I from most of the northeastern species range shows a tendency of recent population expansion, which is evident in Subclades I-A from the northernmost range and I-B from southern Tohoku to northern Kanto and eastern Chubu. In contrast, Subclades I-C from northeastern Chubu and Sado Is. and I-D from northwestern Chubu to Hokuriku seem to have been relatively stable in population size. Hynobius nigrescens differs greatly from other salamander species from northeastern Japan in its much more recent periods of genetic differentiation and its pattern thereof, and is suggested to be a young faunal element in this region.

Recently, two mitochondrial haplotypes, H4 and H8, of Manis sp. were found in two seizures in Hong Kong that do not correspond to Manis javanica, Manis pentadactyla or Manis crassicaudata of Asian pangolin species or any African pangolin species. It was proposed that both haplotypes derived from Manis culionensis, an unknown lineage of M. javanica, or a thus far unidentified Asian pangolin species (Manis sp.). To further investigate these three hypotheses, we used two mitochondrial genes of all eight known extant pangolin species and conducted phylogenetic tree reconstructions, divergence time estimation, and species delimitation analyses. All analyses consistently confirmed that these two haplotypes of Manis sp. constitute a distinct lineage, potentially representing a fifth Asian pangolin species, which originated around the Late Miocene to Early Pliocene (6.95 [4.64–9.85] million years ago). Our study provides genetic support for a potential fifth Asian pangolin species and helps to better understand species diversity of Asian pangolins, which is urgently needed for effective conservation work.

In Hiroshima Bay, parasitic isopods of the genus Mothocya infest the black sea bream Acanthopagrus schlegelii (Bleeker, 1854) and the Japanese halfbeak Hyporhamphus sajori (Temminck and Schlegel, 1846), two fish species that are abundant and commercially important in the Seto Inland Sea of Japan. Immature and mature Mothocya individuals can infect both juveniles and adults of H. sajori, while immature Mothocya are known to parasitize juveniles of A. schlegelii; i.e., no Mothocya parasites are found in adult A. schlegelii. The identification of the immature Mothocya parasitizing juveniles of A. schlegelii remains uncertain, because Mothocya species are morphologically identifiable only based on adult females. Also, the biological/ecological relationship between the hosts and parasites has not been studied. Here, we identified the parasites on A. schlegelii as Mothocya parvostisBruce, 1986 by molecular sequence analyses along with other parasites obtained from H. sajori, the latter being morphologically confirmed by comparison with paratype materials of M. parvostis as well as the similar congener Mothocya sajoriBruce, 1986. The growth rates of the infected A. schlegelii juveniles from June to September in the years 2013–2015 and 2018 were significantly lower than those of the uninfected ones, suggesting a negative effect of the infection on the hosts. Our data on the prevalence and duration of the infection, as well as the body size gain of the hosts and parasites, corroborate a hypothesis that M. parvostis would utilize A. schlegelii as an optional intermediate host before it reaches the final host, H. sajori.

To satisfy their requirements for food and safety, animals need certain habitats to live. Marmots generally select habitats with certain elevation, land surface temperature, soil and vegetation type, and certain mountain slope and aspect; however, what habitats are needed at relatively smaller scales are poorly known. The Himalayan marmot (Marmota himalayana) is distributed mainly on the Qinghai-Tibet Plateau, a region exhibiting diversified topographic features, and the Zoige wetland in the northeast part of the plateau is also the home of the Himalayan marmot. The region is famous for its plateau peat bog, and the suitable habitats for Himalayan marmots are patchily distributed in the wetland. To investigate what kinds of patches are preferred by the marmot in this wetland ecosystem, we measured and compared the soil and vegetation characteristics of used and unused patches. We found that unlike factors governing the habitat selection at macroscales, patches characterized by flat ground and low soil moisture content, with medium vegetation standing height and low vegetation density, are selected in the Zoige wetland. Patches of this kind are selected to meet the marmots' requirements for burrow construction and predator avoidance in such a wetland ecosystem. Together with previous studies on habitat selection of the marmot species at macroscales, we showed that to explore how the animals survive in an environment, it is important to conduct the analysis at multiple scales.

Amphibians exhibit phenotypic plasticity, which allows flexible adaptation to fluctuating environments. Although genes involved in expression of plastic phenotypes have been identified, the endocrine bases of plastic responses are largely unknown. Larvae of the Hokkaido salamander (Hynobius retardatus) plastically display distinct phenotypes, an “offensive phenotype” characterized as larger body with broadened gape and a “defensive phenotype” characterized as enlarged gills and tail and less active behavior, in the presence of prey larval amphibians and predatory larval dragonfly, respectively. In the presence of both prey and predators, the degree of induction of both phenotypes is reduced, suggesting cross-talk between the molecular signaling pathways of these phenotypes. We conducted a transcriptomic analysis to examine how endocrine regulation affects the phenotypic expression by focusing on the pituitary gland. We found that five endocrine genes, i.e., calcitonin related polypeptide alpha (CALCA), growth hormone (GH), neuropeptide B (NPB), parathyroid hormone 2 (PTH2), and prolactin 1 (PRL1), were involved in the expression of both phenotypes. However, we conducted only RNA-seq analysis, and no confirmation of significant up-regulation or down-regulation has been conducted. These results suggest that these genes were up-regulated for induction of the offensive phenotype and down-regulated for induction of the defensive phenotype. Phylogenetic analysis indicated that possible gene duplications of PRL and CALCA have occurred during amphibian evolution. Based on these findings, it is suggested that a trade-off of molecular signaling pathways exists between the two distinct phenotypic expressions. The results also suggest that hormonal-gene duplications might have contributed to the acquisition of phenotypic plasticity in amphibians.

The morphology and morphometry of the olfactory organ of Korean torrent catfishes, genus Liobagrus, consisting of only five endemic species, were investigated by stereo microscopy, scanning electron microscopy, and statistical analysis. They showed the same morphological structure, externally and internally, including a tubular anterior nostril, a slit posterior nostril, and a rosette structure with several linguiform lamellae. Interestingly, however, the lamellar number (LN) revealed specific characteristics useful to identify the five species anatomically: 16–19 in Liobagrus andersoni (with standard length, SL, 96.8 ± 5.5 mm, mean ± SD), 14–16 in Liobagrus obesus (86.9 ± 13.4 mm), 22–27 in Liobagrus mediadiposalis (99.8 ± 14.7 mm), 19–24 in Liobagrus somjinensis (90.1 ± 6.7 mm), and 14–18 in Liobagrus hyeongsanensis (74.0 ± 6.7 mm). Regarding SL, that of L. andersoni was longer than those of L. somjinensis and L. hyeongsanensis. As opposed to the SL, the LN to SL ratios of L. somjinensis (24.1 ± 2.1%) and L. hyeongsanensis (21.1 ± 1.4%) were greater than that of L. andersoni (18 ± 1.2%). These differences might be considered to reflect an interspecific morphological adaptation to micro-habitat according to olfactory importance and can be used as a taxonomic characteristic for this genus.

Nest predation risk is an important factor that may promote the evolution of adaptive parental reproductive strategies in animals. In altricial birds, where parents feed their offspring at the nest for a period of time, parent birds must balance the benefit from increased nest visits and the cost from increased nest predation resulting from their nest visits being detected by predators. Empirical evidence has shown a relationship between parental nest visits and nest predation risks, and parents have been found to be able to reduce the chance of being detected by predators by synchronizing their feeding visits while maintaining feeding rate unchanged. However, it remains poorly understood whether the relationships are universal to all species. We investigated whether nest survival is correlated with parental feeding rate and feeding synchrony in the silver-throated tit (Aegithalos glaucogularis), a songbird endemic to China. We found that parental feeding rate varied by brood size, nestling age and the time during the day, but feeding synchrony was unrelated to the above factors. Moreover, we showed no effect of parental feeding rate or synchrony on nest survival. The failure to find a relationship between parental feeding synchrony and nest survival seems exceptional because it is contrary to findings of previous studies. While there was likely a publication bias regarding previous studies, it is also possible that the evolution of feeding synchrony in different species has different explanations, or the complexity of the predator community and other factors affecting nest survival need to be considered in the analyses.

The red-bellied form of Calliophis intestinalis (Laurenti, 1768) sensu lato was originally reported from Pahang, west Malaysia. To determine the taxonomic status of this form, we examined the type specimens of Elaps sumatranusLidth De Jeude, 1890, Calliophis intestinalis everetti (Boulenger, 1896), and Callophis furcatus var. nigrotaeniatusPeters, 1863. The results indicated that the red-bellied form of C. intestinalis should be named as Calliophis nigrotaeniatus comb. nov., whose valid species status was based on morphological and molecular analyses. We designate a lectotype and redescribe the species, which is genetically close to Calliophis bilineatus (Peters, 1881) from the Philippines, and is clearly distinguishable from other congeners by possessing a pair of gray or dark blue lateral stripes and by being bright red on the ventrum. Elaps sumatranus and C. i. everetti are relegated to subjective junior synonyms of C. nigrotaeniatus.