One-egg twins, in general, initiate embryonic development at the same time, and their developmental stages proceed in parallel. Here we report a rare case of the embryonic development of the red-eared slider turtle, Trachemys scripta, in which twins at conspicuously different developmental stages developed on a single yolk. One of the twins appeared to have developed at the normal developmental rate, whereas the development of the other was markedly delayed, despite the absence of any overt anomalies. This observation suggests uncoupled or fully independent differential regulation of embryonic development from either a single or, more likely, two distinct pluripotent blastoderms sharing the same yolk and same environmental conditions.

The grey-faced buzzard (Butastur indicus) is a raptor that inhabits East Asia, including Japan. Because the number of individuals has decreased by 75% over the last 40 years, this species is classified as vulnerable (VU) in Japan. In the present study, wesought to reveal the genetic structure of the Japanese grey-faced buzzard population at several breeding sites, and to assess the levels of genetic diversity within the Japanese population. We sequenced 555 bp of the mitochondrial DNA of 96 individuals sampled during the breeding season at 18 sites, and 11 individuals sampled during the winter season at one site. In total, 21 variable sites were found in the control region, and we detected 26 haplotypes among the 107 individuals. Fukuoka represented the core breeding area for grey-faced buzzards, as half of all haplotypes were detected there. Four unique haplotypes were detected in the overwintering area. The results of the network and mismatch distribution analyses indicated that the grey-faced buzzard has not experienced a genetic bottleneck in the past, but did experience recent population expansion. In addition, comparisons with other raptors revealed rich genetic diversity in the grey-faced buzzard population. Our results indicate that conservation of both breeding and wintering areas is important for the protection of the grey-faced buzzard.

In the agouti signaling gene protein (Asip) of the house mouse (Mus musculus), inverted repeat (IR) arrays are known to exist in a non-coding region adjacent to the ventral-specific promoter region and the accompanying two exons (exons 1A and 1A′), which are around 100 kb upstream from the amino acid coding regions of exons 2, 3, and 4. To determine the gene structure of mammalian Asip and to elucidate trends in its evolution, non-coding sequences of six rodent (mouse, rat, Chinese hamster, squirrel, guinea pig, and naked mole rat) and three non-rodent (rabbit, human, and cow) species were retrieved from databases and compared. Our homology search analyses revealed the presence of three to five highly conserved non-coding elements (CNE). These CNEs were found to form IRs in rodents and lagomorphs. Combinations of IRs were further shown to build symmetric, long IR arrays. Intra- and inter-specific comparisons of the sequences of three universal CNEs showed homogeneity between CNE pairs within species. This implies that certain evolutionary constraints maintained the IR structure in the rodent and rabbit species.

The use of physical barriers is a common defensive strategy in small-sized endophagous arthropods, but this feeding mode often results in tracks being left on host organisms, thus increasing predation risk. Mechanisms of escape from tracking predators are thus particularly important for endophagous arthropods. Leaf miners are herbivorous insects that inhabit the interiors of leaves and produce various forms of tracks on their host plants. Such tracks are called “mines,” and parasitoid wasps, which are the primary enemy of leaf miners, use mines as cues to find host larvae. In the present study, we use the leaf-mining moth Acrocercops transecta (Insecta: Lepidoptera: Gracillariidae), which changes mine forms during larval growth, and its primary parasitoid Aneurobracon philippinensis (Hymenoptera: Braconidae). Larvae of A. transecta make narrow linear mines in the first and second instars, the third instars expand the mines to flat blotch mines, and the fourth and fifth instars construct three-dimensional tentiform blotch mines. A laboratory parasitization experiment showed that successful oviposition rates were significantly lower on tentiform blotch mines than on other mine types. In contrast, all fifth instars that were transplanted into flat blotch mines were oviposited, suggesting that older instars did not deter ovipositing parasitoids and that the lower rates of successful oviposition on tentiform blotch mines were attributable to refuges inside such mines provided by their three-dimensional structure. Field data demonstrated a plateau in parasitism rates in fourth instar larvae, confirming the results of the laboratory experiment. These results indicate that different mine forms affect the viability of endophagous larvae.

The growth rate, reproduction, recruitment and feeding of four spatangoid species in the Okiislands in the Japan Sea were investigated over five years. Nacospatangus alta, which inhabits unstable surface sediments, grows rapidly, reaches sexual maturity early, and has a short life span, indicating that it should be a ruderal, whereas Metalia spatagus and Brissus agassizii, which inhabit relatively stable deep sediment, grow slowly, reach sexual maturity late, and have a long life span, suggesting that they are stress-tolerators. Lovenia elongata, however, inhabits unstable surface sediment but has an exceptional life history; it grows rapidly, but does not reach sexual maturity early and has a long life span, likely because the specific morphology of its spines and tubercles allow it to cope with surface disturbances caused by storms. Lovenia elongata seems to be a competitive ruderal. A trade-off between test formation and gonad development may occur; N. alta constructs a fragile test with very thin plates, allowing the echinoid to allocate energy to increasing test size and developing the gonad to sexual maturity within a year. Lovenia elongata, with thick plates supporting the specific stout spines and tubercles, needs 2 years to reach sexual maturity with a similar rate of test growth to that of N. alta; M. spatagus and B. agassizii construct robust tests with thick plates, presumably necessary for these species, which burrow and live deep in sand under high pressure from surrounding sand. These echinoids do not reach sexual maturity until over 2 years of age. The flexible trade-off related to stress and disturbance associated with burrowing depth in different habitats allows the spatangoids to have different life-history strategies.

Spiral cleavage is a mode of embryonic cell division found in species from several Phyla, including molluscs, annelids and flatworms. It reflects a tilting in the direction of spindle orientation and cell division at the 4 to 8-cell stage, which may be dextral or sinistral, and propagates into later organismal asymmetry. Genetic analysis in a small number of gastropod molluscs shows the direction of spiral cleavage is determined by maternal genotype, though whether this is also the case more generally for spiralians, and whether spiral cleavage at the 4–8 cell stage is preceded by earlier internal chirality in any spiralian species, is unknown. Here we study the early cleavage stages of two equal-cleaving spiralians, the dextral annelid Spirobranchus lamarcki and the sinistral mollusc Biomphalaria glabrata, using light sheet microscopy to image subcellular vesicles in live embryos and asking if chirality of movement is identifiable. We observe variability in the early cleavage of S. lamarcki, including a viable 3-cell stage. Image data are analysed by both particle tracking and particle image velocimetry. Neither finds evidence for chiral movement in 1-, 2-, 3-, or 4-cell embryos, nor do we detect consistent differences between the embryos of the dextral and sinistrai species. The methodological and evolutionary implications of this are discussed.

The cosmopolitan littoral rotifer Euchlanis dilatata is a potential model species in eco-toxicological and ecological studies, owing to its unique characteristics, such as periphytic benthic living, sensitivity to toxicity, and laying eggs on substrates instead of carrying them on the mother's body. However, studies about the life history strategy of this species, in contrast to the intensely studied planktonic Brachionus species, are relatively scarce. Temperature is an important factor that affects the rotifer's life history traits, and is well known to induce phenotypic plasticity in planktonic rotifers. The present study measured life table parameters of the rotifer E. dilatata originating from one clonal population local to Beijing under three temperature treatments: 14°C, 20°C and 26°C (10 females/replicate × 4 replicates for each treatment). We also examined trade-offs between egg size and egg number, as well as current reproduction and future survival to evaluate the life history strategy of the rotifer. In contrast to cold conditions, in which mothers produced a few large eggs with similar egg sizes, in warm environments, mothers produced many smaller eggs and a few larger eggs, thus resulting in a clear variation in egg sizes. With increasing temperature, the pre-mature period, reproductive period, life span and generation time shortened, while the net reproductive rate and population growth rate increased. Temperature also affected the trade-off between current reproduction and future survival. This work provides basic data for the potential model species E. dilatata.

Here, we demonstrated an antagonistic effect of short neuropeptide F (sNPF) in modulating feeding motivation in the silkworm Bombyx mori; sNPF reduced the feeding-delaying effects caused by administration of an inhibitory peptide, allatotropin (AT). In situ hybridization and MALDI-TOF MS analysis revealed the presence of three subtypes of sNPFs (sNPF-1, -2, and -3) in the midgut enteroendocrine cells. Ca²⁺-imaging analyses revealed that three subtypes of sNPF receptors (sNPFRs) (BNGR-A7, -A10, and -A11) showed different affinities with the three subtypes of sNPFs. In addition, sNPF activated its signaling via ERK phosphorylation in the midgut, while mixture of sNPF and AT reduced the phosphorylation level, agreeing with the results of behavioral assay. Together, our current findings suggest that intestinal sNPF positively modulates the feeding motivation by reducing the inhibitory effects by AT within the midgut.

The cathelicidins represent an important family of host defense peptides (HDPs) found exclusively in vertebrates, which serve as a critical effector in host immune response against microbial infections. To date, a large number of cathelicidins has been identified from diverse vertebrates, such as mammals, birds, reptiles, amphibians and fishes. Sixteen cathelicidins have been identified from snakes in the Elapidae, Viperidae, and Biodae families. However, no cathelicidin has been discovered from a snake of the Colubrinae family. In the present study, we report the identification and characterization of a novel cathelicidin, SA-CATH, from the Colubrinae family snake, Sinonatrix annularis. The cDNA sequence encoding SA-CATH precursor is 735 bp in length, and the mature peptide (SA-CATH) is composed of 30 amino acid residues. Sequence alignment result indicated that SA-CATH precursor possesses relatively high sequence similarity with the cathelicidins from Elapidae and Viperidae family snakes. Similar as the cathelicidins from Elapidae, Viperidae, and Biodae family snakes, SA-CATH mainly assumes an amphipathic alpha-helical conformation, and possesses potent antimicrobial, biofilm inhibitory and anti-inflammatory activities. The results in the present study imply that cathelicidins serve as a kind of conserved effectors with similar structures in the immune systems of Colubrinae, Boidae, Elapidae and Viperidae family snakes. The identification of SA-CATH provides novel clues for the understanding of function and evolution of snake immune systems. The potent antimicrobial, biofilm inhibitory, anti-inflammatory, and slight cytotoxic activities of SA-CATH imply that it is a potential drug candidate in novel antimicrobial agent development.

The intestinal microbiome is known to affect host health through various effects on nutrition and immunity. The oriental honey buzzard (OHB) is a raptor that feeds on bees and wasps. Due to its restricted diet, we reasoned that the OHB may have a unique microbiome. The aim of this study was to characterize the structure of the intestinal flora of oriental honey buzzards and to investigate the difference of intestinal bacterial flora between individuals in the wild and those reared in captivity. We investigated the intestinal microbiome of seven wild buzzards (Wild), one zoo-reared (Zoo), and one individual reared in captivity for one month (Rearing). Average operational taxonomic units in Wild and Rearing were 69.4 and 113, respectively. Diversity indices such as ACE, Chao 1, Shannon, and Alpha were significantly lower in the Wild than in the Rearing samples. These results suggest that the variety of Wild microbiome is remarkably low. At the phylum level, the composition of the microbiome was similar in all three groups, with firmicutes and bacteroidetes predominating. The third most abundant bacterium in Wild was Proteobacteria, whereas it was Actinobacteria in Rearing and unclassified bacteria in Zoo. Thus, microbiome composition is affected even with just one month of human rearing.

To elucidate the genetic population structure of two coastal weevils, Scepticus griseus and S. tigrinus, we conducted molecular phylogenetic analyses of the mitochondrial DNA cytochrome c oxidase subunit I (COI) region (1308 bp) and cytochrome c oxidase subunit II (COII) region (584 bp). A total of 650 individuals (S. griseus, 444 individuals; S. tigrinus, 206 individuals) were obtained from 64 sites. The haplotype networks of both species showed three major lineages with roughly regional distribution. However, the two species show quite different genetic structures; S. griseus has a complicated structure while that of S. tigrinus is simple. We hypothesize that the genetic structure of each of these two weevil species reflects climatic oscillations during the Pleistocene, and the differences in genetic structure between S. griseus and S. tigrinus may represent a unique evolutionary history scenario in each species.