The Pacific bluefin tuna, Thunnus orientalis, is a highly migratory species that is widely distributed in the North Pacific Ocean. Like other marine species, T. orientalis has no external sexual dimorphism; thus, identifying sex-specific variants from whole genome sequence data is a useful approach to develop an effective sex identification method. Here, we report an improved draft genome of T. orientalis and male-specific DNA markers. Combining PacBio long reads and Illumina short reads sufficiently improved genome assembly, with a 38-fold increase in scaffold contiguity (to 444 scaffolds) compared to the first published draft genome. Through analysing re-sequence data of 15 males and 16 females, 250 male-specific SNPs were identified from more than 30 million polymorphisms. All male-specific variants were male-heterozygous, suggesting that T. orientalis has a male heterogametic sex-determination system. The largest linkage disequilibrium block (3,174 bp on scaffold_064) contained 51 male-specific variants. PCR primers and a PCR-based sex identification assay were developed using these male-specific variants. The sex of 115 individuals (56 males and 59 females; sex was diagnosed by visual examination of the gonads) was identified with high accuracy using the assay. This easy, accurate, and practical technique facilitates the control of sex ratios in tuna farms. Furthermore, this method could be used to estimate the sex ratio and/or the sex-specific growth rate of natural populations.
Background Previously, we conducted an epidemiological study screening for Helicobacter pylori antibody positivity among Japanese junior high school students. In this study, we updated the epidemiological data and assessed the clinical features of H pylori antibody‐positive junior high school students. Materials and methods We assessed H pylori antibody‐positive subjects who were identified between 2012 and 2015 at four junior high schools in Nagano Prefecture, Japan. H pylori infection was confirmed by urea breath test (UBT) or endoscopic examination. Endoscopy was performed after obtaining consent from the subject and their guardians. Eradication therapy consisted of triple therapy with proton pump inhibitor (PPI), amoxicillin (AMPC), and clarithromycin (CAM) or metronidazole (MNZ) for seven days. Eradication of H pylori was confirmed by UBT. We reviewed subjects’ characteristics, endoscopic findings, histological findings, eradication regimes, outcomes, and adverse effects. Results The overall prevalence of H pylori antibody positivity was 3.2% (42/1298). We assessed thirteen H pylori antibody‐positive subjects. Eight subjects had a family history of H pylori infection. Six subjects had abdominal pain, and two subjects had iron deficiency anemia (IDA). Twelve subjects underwent endoscopy; one subject had duodenal ulcer, eleven subjects had antral nodular gastritis, and six subjects showed grade 2 closed type atrophic border according to the Kimura‐Takemoto classification. All subjects received eradication therapy; CAM was used in five subjects with CAM susceptibility as well as in three subjects with unknown information on CAM susceptibility, and MNZ was used in five subjects with CAM resistance. Eradication was successful in twelve subjects (one unconfirmed). There were three mild adverse effects (abdominal pain or diarrhea). Conclusions Helicobacter pylori test for junior high school students represents an opportunity to diagnose the peptic ulcer, iron deficiency anemia, and gastric atrophy.
Sex-identification DNA markers are useful tools for sexing organisms that lack externally visible sexual dimorphism, and thus, they provide biological information for ecological and evolutionary studies. Tunas of the genus Thunnus (Scombridae), which comprises 8 species, lack sexual dimorphism of external morphology or coloration. In this study, we applied recently developed genotypic sex-identification markers for Pacific bluefin tuna to other tuna species to evaluate their effectiveness in sex identification. A sex-identification marker named ‘primer pair II’ demonstrated relatively high effectiveness in all tuna species, except southern bluefin tuna. Primer pair II was further tested in 209 albacore individuals collected during the scientific observer program onboard Japanese commercial long-line vessels, and it demonstrated robust performance for genotypic sex identification. The sex ratio of this albacore sample (1:1.4) significantly deviated from the expected 1:1 with the dominance of males, and the mean body size of males was higher than that of females. As all cross-species amplifications of the male-specific markers, except those for the southern bluefin tuna, were male-heterozygous polymorphisms, it is likely that a male-heterozygous sex-associated region exists in the Thunnus genome. The evolution of sex-determination systems in tunas was analyzed by ancestral state reconstruction, which showed that a common ancestor, before the evolution of the genus, possessed the male-heterozygous sex-associated genome region.
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