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Nocardiosis caused by Nocardia seriolae is one of the major threats in the aquaculture of Seriola species (yellowtail; S. quinqueradiata, amberjack; S. dumerili and kingfish; S. lalandi) in Japan. Here, we report the complete nucleotide genome sequence of N. seriolae UTF1, isolated from a cultured yellowtail. The genome is a circular chromosome of 8,121,733 bp with a G+C content of 68.1% that encodes 7,697 predicted proteins. In the N. seriolae UTF1 predicted genes, we found orthologs of virulence factors of pathogenic mycobacteria and human clinical Nocardia isolates involved in host cell invasion, modulation of phagocyte function and survival inside the macrophages. The virulence factor candidates provide an essential basis for understanding their pathogenic mechanisms at the molecular level by the fish nocardiosis research community in future studies. We also found many potential antibiotic resistance genes on the N. seriolae UTF1 chromosome. Comparative analysis with the four existing complete genomes, N. farcinica IFM 10152, N. brasiliensis HUJEG-1 and N. cyriacigeorgica GUH-2 and N. nova SH22a, revealed that 2,745 orthologous genes were present in all five Nocardia genomes (core genes) and 1,982 genes were unique to N. seriolae UTF1. In particular, the N. seriolae UTF1 genome contains a greater number of mobile elements and genes of unknown function that comprise the differences in structure and gene content from the other Nocardia genomes. In addition, a lot of the N. seriolae UTF1-specific genes were assigned to the ABC transport system. Because of limited resources in ocean environments, these N. seriolae UTF1 specific ABC transporters might facilitate adaptation strategies essential for marine environment survival. Thus, the availability of the complete N. seriolae UTF1 genome sequence will provide a valuable resource for comparative genomic studies of N. seriolae isolates, as well as provide new insights into the ecological and functional diversity of the genus Nocardia.

Using a metagenome library constructed from a bacterial associated with a marine sponge Hyrtios erecta, we identified a novel esterase that belongs to the SGNH hydrolase superfamily of esterases. The substrate specificity of EstHE1 was determined using p-nitrophenyl (pNP) ester (C2: acetate, C4: butylate, C6: caproate, C12: laurate, C16: palmitate). EstHE1 exhibited activity against C2 (5.6 U/mg), C4 (5.1 U/mg), and C6 (2.8 U/mg) substrates. The optimal temperature for EstHE1 esterase activity of the pNP acetate substrate was 40°C, and EstHE1 retained 60% of its enzymatic activity in the 30-50°C range. This esterase showed moderate thermostability, retaining 58% of its activity even after preincubation for 12 h at 40°C. EstHE1 also maintained activity in high concentrations of NaCl, indicating that this esterase is salt-tolerant. Thus, EstHE1 has the thermal stability and salt tolerance necessary for use as an industrial enzyme.

Nonagglutinating Lactococcus garvieae has been isolated from diseased farmed yellowtail in Japan since 2012. In this study, the complete genome and plasmid sequence of nonagglutinating L. garvieae strain 122061 was determined, to our knowledge, for the first time.

Giant clams are simultaneous hermaphrodites and are assumed to ejaculate first and, after completely stopping ejaculation, release eggs. In the seed production method aimed at preventing self-fertilization, each adult clam is induced to ejaculate in a tank and then release eggs in another tank. Giant clams, however, have recently been suggested to continue ejaculation for a period after the beginning of egg release. The overlap between ejaculation and egg release might lead to self-fertilization in the tank used for egg release, especially for the eggs released just at the beginning of spawning. We examined the possibility of such self-fertilization for the giant clam Tridacna crocea and obtained three results. (1) In observations with the naked eye in a laboratory, 2 of 38 T. crocea simultaneously ejaculated and released eggs. (2) In a laboratory experiment, 1.5 to 80.0% of eggs released from each adult clam developed into D-shaped larvae without artificial cross-fertilization. Such development occurred more frequently for the eggs released earlier from each adult clam than for the eggs released later from the clam. (3) In observations at a hatchery, 2 to 94% of the eggs released from 4 of 5 adults were found to develop into D-shaped larvae without artificial cross-fertilization. The three results suggest that at least some T. crocea adults continue ejaculation for a period after starting spawning eggs, which causes self-fertilization.

Odontamblyopus lacepedii inhabits burrows in mudflats and breathes air at the surface opening. Investigations of the intertidal burrows using resin casting demonstrated a highly branched burrow system. The burrows are composed primarily of branching patterns of interconnected tunnels and shafts that communicate into two to seven surface openings. Bulbous chambers (i.e., dilated portions of the burrow) at branching sections of the tunnels or shafts are common features of the burrow. The presence of these chambers accords the fish adequate space to maneuver inside the burrow, and thus constant access to the surface. The combination of all burrow characteristics and previously reported variability in air breathing patterns are ostensibly of selective value for aerial predator avoidance during air breathing in O. lacepedii.

Larval settlement rates, genetic structure, and gene flow of broadcast-spawning (Acropora tenuis) and planula-brooding (Stylophora pistillata) corals (Scleractinia) were compared within a 500 km range in the Ryukyu Archipelago. We conducted a laboratory experiment to investigate planula settlement rates, and a broad sampling survey to determine genetic variation in both species in the Archipelago. In the laboratory experiment, the planulae ofS. pistillatasettled a few hours after release, while those ofA. tenuisstarted to settle at least 4 d after the release of gametes. The survival rates and competency periods of larvae were higher and longer forA. tenuisthan forS. pistillata. These results suggest that broader dispersal is more likely forA. tenuisthan forS. pistillata. In the population genetic analysis, we measured local (2 stations in a region) and regional (Okinawa, Kerama and Yaeyama) patterns of genetic variation with allozyme electrophoresis. We also inferred the levels of gene flow in the 2 species. In the study area, gene flow (Nₑm) and genetic distance (D) were, respectively, higher and smaller for the spawnerA. tenuis(Nₑm= 3.5 to 16.4,D= 0.028 to 0.187) than for the brooderS. pistillata(Nₑm= 0.9 to 1.5,D= 0.026 to 0.309). Therefore, the planulae settlement rates were well in agreement with gene flow. In addition, for both species,Nₑmbetween the Okinawa and Kerama regions (30 to 150 km apart;Nₑm= 9.4 to 22.5 inA. tenuisand 1.4 to 3.3 inS. pistillata) was higher than that between the Okinawa-Kerama and Yaeyama regions (up to 500 km apart;Nₑm= 3.1 to 9.4 inA. tenuisand 0.5 to 1.4 inS. pistillata). The results suggest that coral populations in the Kerama Island are a major source of the coral planulae needed for the recovery of both brooding and spawning coral communities around the Okinawa Islands, after the mass-bleaching event in 1998.

The allozymes and morphology of 110 specimens of three Sebastiscus species (S. marmoratus, S. tertius, and S. albofasciatus) in the East China Sea and near Japan were compared. Results of 20 allozyme loci studied showed that all three species were closely related (Nei's unbiased genetic distances, 0.057–0.133) but could be identified on the basis of informative loci with a few exceptions. Initial identification based on color patterns agreed with allozyme identification in more than 98% of Sebastiscus specimens and agreed completely in S. albofasciatus. One specimen that was initially identified as S. marmoratus because of the dark body color was actually S. tertius according to two informative allozyme loci. Number of pectoral fin rays differed between S. marmoratus (18 or fewer, 98%) and S. tertius (19 or more, 85%) in this study. The previously mentioned dark specimen had 19 pectoral fin rays, which are characteristic in S. tertius. Using seven morphological measurements, canonical discriminant analysis between S. marmoratus and S. tertius classified less than 90% of specimens into the original groups (species). Some specimens of S. tertius resembled S. marmoratus in body shape and vice versa. A combination of genetic characterization and morphological examination is necessary to identify S. marmoratus and S. tertius accurately. Distinction based on allozymes and color patterns with numbers of pectoral fin rays should provide satisfactory identification.

We determined the complete nucleotide sequences of the mitochondrial genomes for the three currently recognized species of ocean sunfish: Mola mola, Masturus lanceolatus, and Ranzania laevis (Tetraodontiformes: Molidae). Each genome contained the 37 genes as found in teleosts, with the typical gene order in teleosts. Bayesian. maximum-likelihood, and maximum-parsimony analyses were conducted with the data set comprising concatenated nucleotide sequences from 36 genes (excluding the ND6 gene) of three molids and four outgroups (three tetraodontiforms plus a caproid). The resultant trees supported monophyly of the Molidae and its intrarelationships ((Mola, Masturus), Ranzania), which were congruent with previous morphology-based hypotheses.

Lacking a propensity to emerge over the mud surface, the eel goby, Odontamblyopus lacepedii, survives low tide periods by continuously breathing air in burrows filled with hypoxic water. As with most marine air-breathing fishes, O. lacepedii does not possess an accessory air-breathing organ, but holds air in the buccal-opercular cavity. The present study aimed to clarify how the respiratory vasculature has been modified in this facultative air-breathing fish. Results showed that the gills apparently lacked structural modifications for air breathing, whereas the inner epithelia of the opercula were richly vascularized. Comparison with two sympatric gobies revealed that the density of blood capillaries within 10?m from the inner opercular epithelial surface in O. lacepedii (14.5 ± 3.0 capillaries mm-1; mean ± s.d., n = 3) was significantly higher than in the aquatic non-air-breathing Acanthogobius hasta (0.0 ± 0.0) but significantly lower than in the amphibious air-breathing mudskipper, Periophthalmus modestus (59.1 ± 8.5). The opercular capillary bed was supplied predominantly by the 1st efferent branchial arteries (EBA1) and drained by the opercular veins, which open into the anterior cardinal vein. Deep invaginations at the distal end of the EBA1 and the junction with EBA2 are suggestive of blood flow regulatory sites during breath-holding and apnoeic periods. It remains to be investigated how blood flow through the gills is maintained during breath holding when the buccal–opercular cavity is filled with air.

Genetic and morphological variation were studied in a brooding (ovoviviparous) and morphologically variable freshwater snail (Viviparus georgianus (Lea)) in the southeastern United States. Eleven populations were clustered into three genetically isolated, allopatric species characterized by 7 to 15 diagnostic loci out of the 38 loci examined. These allopatric species were an eastern species (in eastern and southern Florida), a western species (in the Florida panhandle), and a central species in the Ochlockonee River. Nei's standard genetic distances between species were large (0.23-0.52) compared to within-species distances (0.00-0.06). Moreover, genetic distances between the Ochlockonee River species and other species were larger than the distance between the eastern and western species. Hierarchical F-statistics for differentiation among sites within drainage systems (F $\\sb{\\rm SD}$ ) of the western and eastern species were large (0.519 and 0.387, respectively). The F $\\sb{\\rm DT}$values (differentiation among drainage systems within the total area sampled) were negative, so most of the intraspecific genetic differentiation was due to differences among populations within drainage systems, rather than to differences among systems. Canonical discriminant analysis of nine shell measurements separated all three species with little overlap. The type specimens of the Viviparus georgianus complex and type locality specimens were compared to the discriminant function and canonical discriminant analyses of shell characters of the studied samples to assign correct species names. The western species and Ochlockonee River species appear to be Viviparus goodrichi Archer and Viviparus limi Pilsbry, respectively, which were originally described as subspecies of Viviparus contectoides (= Viviparus georgianus). The eastern species is Viviparus georgianus (Lea). The three species can be distinguished by the following morphological characteristics: V. goodrichi has a more globose shell with a larger aperture than V. limi; V. georgianus has shorter aperture height than the other species. Also, these three species can be identified reliably using allozyme characters.