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Psychophysiological studies in vertebrates have focused on taxes as indicators of behavioral change. Actually, a considerable number of studies about anxiety-like and anti-anxiety-like behaviors involving geotaxis, scototaxis, and thigmotaxis have been conducted on fish. However, few analyses considering these behaviors based on taxes in fish have been conducted. Here, using goldfish, we measured the time spent in the bright or dark area of a horizontally long rectangular tank (HLRT), in the upper or lower area of a vertically long rectangular tank (VLRT), and in the central or edge area of a circular tank (CT), respectively, for the first 30 min and the last 30 min in a 3-h period after fish had been introduced to tanks. Dark, lower, and edge preference behaviors were observed for the first 30 min in all tanks. While dark and edge preference behaviors were maintained even for the last 30 min, the lower preference was lost. Swimming distance and the number of area crossings in each tank were also compared between the first 30 min and the last 30 min. Both decreased significantly or tended to decrease in the last 30 min in the HLRT and the CT, but no change was observed in the VLRT. These results suggest that, in goldfish, preference behavior is stable for a short time, and that environmental habituation may depend on the shape of the tank and the elapsed time.
Relaxin-like gonad-stimulating peptide (RGP) is a hormone with gonadotropin-like activity in starfish. This study revealed that spawning inducing activity was detected in an extract of brachiolaria larvae of Patiria pectinifera. Spawning inducing activity in the extract was due to P. pectinifera RGP (PpeRGP), not 1-methyladenine. The expression of PpeRGP mRNA was also found in brachiolaria. Immunohistochemical observation with specific antibodies for PpeRGP showed that PpeRGP was distributed in the peripheral adhesive papilla of the brachiolaria arms. In contrast, PpeRGP was not detected in the adult rudiment or ciliary band regions, which are present in the neural system. These findings strongly suggest that RGP exists in the larvae before metamorphosis. Because gonads are not developed in starfish larvae, it seems likely that RGP plays another role other than gonadotropic action in the early development of starfish.
It has been suggested that aquatic vertebrates may be able to meet their energy requirements by using the amino acids dissolved in environmental water. If this ability can be applied to aquatic organisms generally, then conventional ecological theories related to food web interactions should be revisited, as this would likely bring about significant advances in applications. Here, we prepared two 1 mM amino acid (phenylalanine and glycine) solutions in environmental water and conducted laboratory experiments to demonstrate the utilization of dissolved amino acids by hatchlings of three salamander species (Ezo: Hynobius retardatus, Tohoku: Hynobius lichenatus, and Japanese black: Hynobius nigrescens). Compared to controls (no amino acids in environmental water), the growth rate for Ezo salamanders was higher when larvae were reared in phenylalanine solution, while that for Japanese black salamanders was higher in glycine and phenylalanine solutions. Amino acids in environmental water had no effect on the growth of Tohoku salamanders. However, when growth was divided into early (days 1 to 5) and late (days 5 to 7) developmental stages, growth in early-developmental stage individuals was improved by phenylalanine treatment, even in Tohoku salamanders. The results showed that the growth of salamander larvae was improved when salamanders were reared in environmental water with high amino acid concentrations. Although aquatic bacteria may not have been removed completely from the environmental water, no other eukaryotes were present. Our results suggest an overlooked nutrient pathway in which aquatic vertebrates take up dissolved amino acids without mediation by other eukaryotes.
Marine diatoms are an important food resource for bivalves, but few experimental studies have evaluated diatom assimilation by bivalves. We conducted a laboratory experiment to investigate the ability of the suspension-feeding bivalve Nuttallia olivacea to utilize three common diatom species (planktonic diatoms Thalassiosira pseudonana and Skeletonema dohrnii and the benthic diatom Entomoneis paludosa) as food labeled with heavy nitrogen stable isotope (15N) by incubation in medium containing Na15NO3. The percentage of food-derived nitrogen in the organs of the bivalves increased over time, confirming that the bivalves were taking up dietary nitrogen from diatoms. The proportion of food-derived nitrogen from diatoms to bivalves appeared to be higher in planktonic species than in benthic species. However, it is possible that the benthic diatom intake by the bivalves in this study was underestimated because the substrate was not disturbed as would occur under field conditions. The percentage of food-derived nitrogen in bivalve organs tended to be highest in the digestive diverticula, followed by the foot, mantle, and siphon, regardless of diatom type. These findings suggest that N. olivacea may preferentially distribute nitrogen to organs other than the siphon, which is prone to continuous loss by fish predation.
DNA markers that detect differences in the number of microsatellite repeats can be highly effective for genotyping individuals that lack differences in external morphology. However, isolation of sequences with different microsatellite repeat numbers between individuals has been a time-consuming process in the development of DNA markers. Individual identification of Japanese giant flying squirrels (Petaurista leucogenys) has been challenging because this species is arboreal and nocturnal and exhibits little to no morphological variation between individuals. In this study, we developed DNA markers for sex and individual identification of this species by an efficient method using high-throughput DNA sequence data. Paired-end 5 Gb (2 × 250 bp) and 15 Gb (2 × 150 bp) genome sequences were determined from a female and a male Japanese giant flying squirrel, respectively. We searched SRY and XIST genes located on Y and X chromosomes, respectively, from high-throughput sequence data and designed primers to amplify these genes. Using these primer sets, we succeeded to identify the sex of individuals. In addition, we selected 12 loci containing microsatellites with different numbers of repeats between two individuals from the same data set, and designed primers to amplify these sequences. Twenty individuals from nine different locations were discriminated using these primer sets. Furthermore, both sex and microsatellite markers were amplified from DNA extracted non-invasively from single fecal pellet samples. Based on our results for flying squirrels, we expect our efficient method for developing non-invasive high-resolution individual- and sex-specific genotyping to be applicable to a diversity of mammalian species.
Additional taxonomic and ecological data for the lernaeid copepod Lamproglena hemprichiiNordmann, 1832 infecting the African tigerfish, Hydrocynus vittatus Castelnau, 1861, are presented with scanning electron micrographs, molecular characterization, and selected ecological parameters. Eighty fish were collected from Lake Kariba between October 2014 and July 2015. Scanning electron microscopy provided additional data for the morphology, including structures on the antennulae, antennae, and legs. The 18S and 28S rDNA fragments of this species were distinct from those of other Lamproglena taxa but placed this species within this genus. Phylogenetically, L. hemprichii appears closest to L. monodi Capart, 1944, the only other African species for which molecular data is available. The anterior part of the second gill filament was the preferred attachment site. There was a positive correlation (Pearson correlation coefficient r2 = 0.64; P = 0.77) between the length of the parasite and the length of the fish. A positive correlation (Pearson correlation coefficient r2 = 0.61; P = 0.03) between the length of the parasite and the length of the gill filament was also observed. Although the specimens studied here are morphologically highly similar to previous reports of L. hemprichii, some morphological variation was observed, and a revision (morphometric and genetic) of the taxon is suggested. This study provides the first detailed genetic characterization and phylogenetic information for the species.
Vertical migration as well as horizontal dispersion is important in the ecological strategy of planktonic larvae of sedentary corals. We report in this paper unique vertical swimming behavior of planulae of the reef-building coral Acropora tenuis. Several days after fertilization, most of the planulae stayed exclusively at either the top or the bottom of the rearing tank. A good proportion of the planulae migrated almost vertically between top and bottom with fairly straight trajectories. Planulae sometimes switched their swimming direction via a sharp turn between the opposite directions. Quantitative analyses demonstrated that planulae kept constant speed while swimming either upward or downward, in contrast to frequent changes of direction and speed in horizontal swimming. Statistical comparison of propulsive speeds, estimated from swimming speeds and passive sedimentation, revealed gravikinesis of planulae, where the propulsive speed was significantly greater in downward swimming than upward swimming. The larval density hydrodynamically estimated was 0.25% lower than sea water density, which might be explained by the large quantity of lipids in planulae. Also, the deciliated larvae tended to orient oral end-up during floatation, presumably due to asymmetrical distribution of the endogenous light lipids. Plasticity of the larval tissue geometry could easily cause relocation of the center of forces which work together to generate gravitactic-orientation torque and, therefore, abrupt changing of the gravitactic swimming direction. The bimodal gravitactic behavior may give a new insight into dispersal and recruitment of coral larvae.
Photic entrainment is an essential property of the circadian clock that sets the appropriate timing of daily behavioral and physiological events. However, the molecular mechanisms underlying the entrainment remain largely unknown. In the cricket Gryllus bimaculatus, the immediate early gene c-fosB plays an important role in photic entrainment, followed by a mechanism involving cryptochromes (crys). However, the association between c-fosB expression and crys remains unclear. In the present study, using RNA-sequencing analysis, we found that five Fbxl family genes (Fbxl4, Fbxl5, Fbxl16, Fbxl-like1, and Fbxl-like2) encoding F-box and leucine-rich repeat proteins are likely involved in the mechanism following light-dependent c-fosB induction. RNA interference (RNAi) of c-fosA/B significantly downregulated Fbxls expression, whereas RNAi of the Fbxl genes exerted no effect on c-fosB expression. The Fbxl genes showed rhythmic expression under light-dark cycles (LDs) with higher expression levels in early day (Fbxl16), whole day (Fbxl-like1), or day-to-early night (Fbxl4, Fbxl5, and Fbxl-like2), whereas their expression was reduced in the dark. We then examined the effect of their RNAi on the photic entrainment of the locomotor rhythm and found that RNAi of Fbxl4 either disrupted or significantly delayed the re-entrainment of the locomotor rhythm to shifted LDs. These results suggest that light-induced c-fosB expression stimulates Fbxl4 expression to reset the circadian clock.
Gastropods of over a dozen genera in the family Eulimidae have been identified as parasites of brittle stars, and many more remain to be discovered and described for a comprehensive understanding of the evolutionary history of their host–parasite relationships. In this study, we describe Fusceulimoides kohtsukai gen. et sp. nov., parasitic on the little brittle star, Ophiactis savignyi (Ophiactidae), in Kanagawa, central Japan. The new genus is distinguished from other eulimid genera by the combination of following seven conchological characters: (1) a very small size of up to 1.7 mm high, (2) a colorless translucent appearance, (3) a conical pupiform shape with a paucispiral protoconch, (4) slightly convex teleoconch whorls, (5) a remarkably large body whorl occupying 65–70% of the total shell height, (6) a broad, somewhat squarish and laterally expanded aperture with a strongly curved outer lip, and (7) a developed parietal callus without an indentation or depression in the umbilical area. A multi-locus molecular phylogeny revealed its distant relationship to Hemiliostraca + Pyramidelloides, a previously known clade of ophiuroid parasites, thereby suggesting multiple origins of this host–parasite association in Eulimidae.
Paraedwardsia, a genus of sea anemones within the family Edwardsiidae, is characterized by tenaculi on the surface of the column, often covered by sand grains. Most species of this genus are known from areas deeper than 200 m. From Japanese waters, three specimens were recently collected, and are formally described here as Paraedwardsia rinkaimaruae sp. nov. and Paraedwardsia moroisoensis sp. nov. The latter species was collected from 6 m depth, and this is the shallowest record of this genus. In addition, P. rinkaimaruae sp. nov. has a few sack-like structures in its mesoglea, which might be the key to the evolution of nemathybomes.
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