The Mekong giant catfish Pangasianodon gigas is endemic to the Mekong River basin, and is recognized as endangered species, largely due to overfishing and development of the river basin. We monitored food intake of P. gigas in a stable environment in an aquarium over a 6-year period and analyzed their feeding rhythm and fasting periods. The daily food intake for each fish was recorded from 18 June 2004 to 17 June 2010. The feeding rhythm or pattern was determined by the fast Fourier transform (FFT) analysis. The FFT analysis revealed that different cycles of feeding rhythm (168.8, 313.1, and 365.3 days) in three catfishes and no observable cycles in two catfishes. However, three catfishes showed subordinate peaks with approximately 365 days (365.3 days for all). These suggest that, at least, four of five catfish had have approximately 365-days feeding cycle. We also showed that all catfish undergo long-term fasting periods (> 20 days). Of note, the feeding/fasting pattern coincides with the wet/dry seasons in Thailand, which also corresponds to the abundance of the catfish food resource (Cladophora spp.). We found that P. gigas exhibit a seasonal feeding rhythm that is synchronized by food availability. Furthermore, we found that the seasonal feeding rhythm was gradually dampened over time, suggesting that the observed seasonal feeding rhythm with long-term fasting of the catfish is likely controlled by an endogenous clock system. To our knowledge, this is the first case of quantification of the seasonal feeding rhythm with fasting periods in teleost fish.

A tyrosine-phosphorylated protein of 33 kDa is shown to be present in the solubilized yolk fraction of Xenopus laevis oocytes, eggs, and early embryos. The phosphoprotein is lipovitellin 2 as demonstrated by immunoprecipitation and mass spectrometry studies, and is termed pp33/LV2. Sub-cellular fractionation and immunoblotting studies demonstrate that pp33/LV2 is stably present in the Triton X-100-resistant and SDS-soluble yolk fractions during oogenesis, oocyte maturation, and early embryogenesis. From after the swimming tadpole stages (stage 39∼), however, it becomes partly soluble to Triton X-100-containing buffer and all disappear thereafter (stage 48∼49). In vitro enzyme assays with the use of the tyrosine phosphatase LAR and the tyrosine kinase Src demonstrate the reversible nature of the tyrosine phosphorylation of pp33/LV2. Microinjection studies demonstrate that the solubilized yolk fractions, but not those immunodepleted of pp33/LV2 or those pretreated with LAR, inhibit progesterone- or insulin-induced oocyte maturation. A pp33/LV2-like protein seems to present in two Xenopus subspecies, one other frog species, and two fish species, but not in other amphibian species, such as newt and salamander. These results suggest that LV2, in its tyrosine-phosphorylated form, serves in a cellular function in a species-specific manner, but the mechanism is still unknown.

We examined nucleotide changes that underlie coat color variation in Black Rats (the Rattus rattus species complex), which show polymorphism in dorsal fur color, including either grayish brown (agouti) or black (melanistic) forms. We examined the full coding sequence of a gene known to produce melanism in other vertebrates—melanocortin-1-receptor gene Mc1r (954 bp) —using samples of both R. rattus (with 2n = 38) and its close relative Asian Black Rat (R. tanezumi; 2n = 42). We used 61 specimens from Japan with karyotype-known individuals and four samples from Pakistan. We found 11 allele sequences and constructed a network tree that shows two distinct clusters, with allelic segregation according to karyotype and by inference, representing the two species. We found that a nucleotide substitution from G to A at site 280, producing an amino acid change from glutamic acid to lysine, was associated with the dominant trait of the melanistic form of the coat color in R. rattus. Notably, the derived SNP 280A was found in a single allele, with the ancestral SNP 280G present in seven alleles. By contrast, all three alleles for R. tanezumi retain the ancestral SNP 280G. These results suggest a possible recent origin of melanism in R. rattus.

Marine lakes in the Palau Islands are known to harbor unique marine fauna that have remained isolated since the formation of the lakes after the Last Glacial Maximum. We analyzed mussels from marine lakes located on different islands and conducted morphological, phylogenetic and population genetic characterization to clarify their evolutionary history. The mussels were morphologically classified into three differentiated morphs: NS, ON, and MC. Their common characteristics were consistent with the Brachidontes—Hormomya complex of the Mytilidae family. Phylogenetic analysis based on the nuclear 18S ribosomal RNA gene supported the taxonomic position of the mussels among the Mytilidae. In the mitochondrial cytochrome c oxidase subunit I (COI) gene lineage, NS-and MC-morphs were highly diverged from each other; their estimated time of divergence dates back to the mid-Pleistocene. ON-morph was more closely related to MC-morph, although the shell morphologies of ON- and MC-morphs were easily distinguishable. Population genetic analysis revealed the coexistence of highly diverged haplotypes within a population of ON-morph, indicating introgression of mtDNA among the morphs. Our data suggest that morphological differentiation of marine lake mussels can occur in a relatively short period under different environmental conditions. Thus, the marine lakes provide a unique site for the study of diversification in mussels.

The pine marten, Martes martes, is a medium-sized terrestrial carnivore associated with woodland habitats of the western Palearctic region. The present distribution area of the species also includes six islands of the western Mediterranean basin. The origin of these insular populations and their taxonomic status are still debated; their molecular characterization appears relevant for conservation purposes. To describe the genetic variability of the pine martens from Sardinia we characterized 40 insular and 14 Italian individuals at seven nuclear microsatellite loci. The identification of private alleles and the calculated F_ST value of 0.074 revealed some genetic differentiation between the two populations, which accounts for the high percentages of correct allocation (96.39–98.80%) scored by the genotype assignment test. The presence of two distinct clusters corresponding to Sardinia and mainland Italy was further confirmed by the multivariate Factorial Correspondence Analysis of individual genotypes. Moreover, the genome of the Sardinian individuals bore signs of past demographic fluctuations, i.e. the presence of the monomorphic locus Ma-4, a lower allelic richness and a lower number of private alleles, which may derive from the combination of drift, founder effects, and human overexploitation. Anyway, if such events ever affected the Sardinian population, this is likely to have happened in the past since, according to our microsatellite data, the present-day population does not show evidence of recent bottlenecks or inbreeding, the Wilcoxon sign-rank test and the F_IS index being not statistically significant (both P > 0.05). Based on this genetic evidence, we advance hypotheses about the distinctiveness of the Sardinian population and its significance for taxonomy and conservation.

Previously unreported males of a gnathiid isopod were found in reproductive aggregations of the harem-forming gnathiid Elaphognathia discolor. Although the male gnathiids were small in size and morphologically different from E. discolor males, the male sexual organ, appendix masculina, was similar to that of E. discolor males, and possible conspecific larvae and females of the small male gnathiid were never found. In the laboratory, the small male gnathiids as well as male E. discolor successfully copulated with female E. discolor, and the development of embryos in female brood pouches was observed. Offspring of small male gnathiids develop to adults of E. discolor after molting three times, or small male gnathiids after molting two times. Thus, the small male gnathiid was concluded to be an alternative male form compared to the regular large male form of E. discolor. This male polymorphism was thought to have a genetic basis, since no small male specimens appeared in offspring of regular E. discolor males. Field sampling showed that a regular large male formed a harem composed of one large male and several females and never coexisted with other large males as previously reported. However, small males were often found together with large males. Therefore, small males are thought to be sneakers intruding into harems dominated by large males.

To determine the habitat usage and movement patterns of the viviparous aquatic snake Oocatochus rufodorsatus (formerly Elaphe rufodorsata), we radio-tracked 21 snakes on agricultural lands during two active seasons in 2007 and 2008. Male and female snakes stayed close to aquatic habitats such as paddy fields and agricultural ponds during both breeding and non-breeding periods, except when the snakes moved to dry terrestrial areas to hibernate in late fall. The use of different structural features in the habitat, such as ground, tree, underground, and water, varied depending on the air and water temperatures, female's reproductive conditions, and the time of day. Male and female snakes moved about 17 m daily and postpartum females moved farther than antepartum females. The home ranges of males and females were 0.45 ha and 0.47 ha, respectively, and the year-round home range of this species was approximately 1.54 ha (95% fixed kernel). Thus, to conserve a population of O. rufodorsatus in our study area, areas including both aquatic and terrestrial habitats within a radius of 150 m from a core pond habitat must be preserved.

Species-specific chemical signals released through urine, sweat, saliva and feces are involved in communication between animals. Urinary biochemical constituents along with pheromones may contribute to variation across reproductive cycles and facilitate to estrus detection. Hence, the present study was designed to analyze such biochemical profiles, such as proteins, carbohydrates, lipids, fatty acids, in response with steroid hormones such as estradiol and progesterone. The experimental groups were normal, prepubertal, ovariectomized, and ovariectomized with estrogentreated female mice. In normal mice, the protein and lipid concentrations in urine were significantly higher in proestrus and estrus phases and the quantity of fatty acids was also comparatively higher in estrus. Furthermore, certain fatty acids, namely tridecanoic, palmitic and oleic acids, were present during proestrus and estrus phases, but were exclusively absent in ovariectomized mice. However, the carbohydrate level was equally maintained throughout the four phases of estrous cycle. For successful communication, higher concentrations of protein and specific fatty acids in estrus are directly involved. The significant increase in estradiol at estrus and progesterone at metestrus seems to be of greater importance in the expression pattern of biochemical constituents and may play a notable role in estrous cycle regulation. Thus, we conclude that the variations observed in the concentration of the biochemical constituents depend on the phase of the reproductive cycle as well as hormonal status of animals. The appearance of protein and specific fatty acids during estrus phase raises the possibility to use these as a urinary indicators for estrus detection.

Following FIASCO protocol and across-amplification approach, we present eleven microsatellite primers of the Oriental White stork, Ciconia boyciana in this article. All loci were polymorphic, except for locus Cbo235, which possessed two alleles but was homozygous in all 23 individuals of C. boyciana used in this study. The number of the alleles per locus ranged from two to eight, and the observed heterozygosity (H_o) and expected heterozygosity (H_E) ranged from 0 to 0.857 and 0.222 to 0.851, respectively. These markers are proved useful in genetic study of C. boyciana.

Insulin family peptide members play key roles in regulating growth, metabolism, and reproduction. Bombyxin is an insulin-related peptide of the silkmoth Bombyx mori. We analyzed the full genome of B. mori and identified five novel bombyxin families, V to Z. We characterized the genomic organization and chromosomal location of the novel bombyxin family genes. In contrast to previously identified bombyxin genes, bombyxin-V and -Z genes had intervening introns at almost the same positions as vertebrate insulin genes. We performed reverse transcription-polymerase chain reaction and in situ hybridization in different tissues and developmental stages to observe their temporal and spatial expression patterns. The newly identified bombyxin genes were expressed in diverse tissues: bombyxin-V, -W, and -Y mRNAs were expressed in the brain and bombyxin-X mRNA in fat bodies. Bombyxin-Y gene was expressed in both brain and ovary of larval stages. High level of bombyxin-Z gene expression in the follicular cells may suggest its function in reproduction. The presence of a short C-peptide domain and an extended A chain domain, and high expression of bombyxin-X gene in the fat body cells during non-feeding stages suggest its insulin-like growth factor-like function. These results suggest that the bombyxin genes originated from a common ancestral gene, similar to the vertebrate insulin gene, and evolved into a diverse gene family with multiple functions.

The importance of the tongue during feeding, and the limited information on the tongue of most aquatic mammals led us to investigate its morphological aspects in sexually immature and mature Sotalia guianensis. Six tongues were measured and photo-documented after their removal from the oral cavity. The samples were divided into rostral, middle, and caudal regions, and examined using light microscopy and scanning electron microscopy (S.E.M.). Sotalia guianensis tongue presented lateral grooves from the apex to the middle portion, while the anterolateral region presented marginal papillae. Histological characteristics revealed the presence of a keratinized stratified epithelium, salivary glands in the middle and caudal portions of the tongue, and filiform papillae in the caudal region. S.E.M. images revealed the presence of filiform papillae and ducts of salivary glands in the middle and caudal portions of the tongue. We can conclude that the characteristics found in this study may reflect an adaptation to changes in diet after weaning.