There has been a long-standing controversy about whether vertebrates emerged in the Paleozoic from marine or freshwater environments. Several hypotheses have proposed coastal, estuarine and riparian areas as sites of the transition. Here, we report the ecology of an amphibious fish Periophthalmodon septemradiatus, which we presume is in the process of niche expansion into terrestrial habitats from estuarine to freshwater environments along the Mekong River, Vietnam. Adult fish are highly terrestrial and have not been observed to venture into water during our survey. Courtship behaviour was observed, and fertilised eggs were recovered from burrows in both brackish and freshwater environments. The smallest fish collected at 12, 96, and 148 km from the river mouth were juveniles shortly after starting an amphibious life. These findings suggest reproduction in both brackish and freshwater environments. In contrast, otolith Sr:Ca ratio indicates larval hatching only in brackish water. Analysis of a 940-base pair (bp) segment of the mitochondrial cytochrome c oxidase subunit II and a 934-bp segment of the mitochondrial D-loop demonstrated no genetic segregation between populations. The fish may provide a unique opportunity to study how ambient salinity affects the biology and ecology of a living vertebrate during transition from water to land.

The species classification of Cambodian freshwater pufferfish is incomplete and confusing, and scientific information on their toxicity and toxin profile is limited. In the present study, to accumulate information on the phylogeny and toxin profile of freshwater pufferfish, and to contribute to food safety in Cambodia, we conducted simultaneous genetic-based phylogenetic and toxin analyses using freshwater pufferfish individuals collected from Phnom Penh and Kratie (designated PNH and KTI, respectively). Phylogenetic analysis of partial sequences of three mitochondrial genes (cytochrome b, 16S rRNA, and cytochrome c oxidase subunit I) determined for each fish revealed that PNH and KTI are different species in the genus Pao (designated Pao sp. A and Pao sp. B, respectively). A partial sequence of the nuclear tributyltin-binding protein type 2 (TBT-bp2) gene differentiated the species at the amino acid level. Instrumental analysis of the toxin profile revealed that both Pao sp. A and Pao sp. B possess saxitoxins (STXs), comprising STX as the main component. In Pao sp. A, the toxin concentration in each tissue was extremely high, far exceeding the regulatory limit for STXs set by the Codex Committee, whereas in Pao sp. B, only the skin contained high toxin concentrations. The difference in the STX accumulation ability between the two species with different TBT-bp2 sequences suggests that TBT-bp2 is involved in STX accumulation in freshwater pufferfish.

The complete mitochondrial genome of Anisakis pegreffii (former A. simplex A) was determined using the Illumina HiSeq platform. The genome was 14,002 bp in length made up of 36 mitochondrial genes (12 CDSs, 22 tRNAs, and 2 rRNAs). Phylogenetic analysis clarified the mitogenome sequences of the three sibling species of the A. simplex species complex, A. pegreffii, A. simplex sensu stricto and A. berlandi (former A. simplex C).

Marine Takifugu pufferfish, which naturally possess tetrodotoxins (TTXs), selectively take up and accumulate TTXs, whereas freshwater Pao pufferfish, which naturally possess saxitoxins (STXs), selectively take up and accumulate STXs. To further clarify the TTXs/STXs selectivity in pufferfish, we conducted a TTX/STX administration experiment using Chelonodontops patoca, a euryhaline marine pufferfish possessing both TTXs and STXs. Forty nontoxic cultured individuals of C. patoca were divided into a seawater group (SW, acclimated/reared at 33‰ salinity; n = 20) and a brackish water group (BW, acclimated/reared at 8‰ salinity; n = 20). An aqueous TTX/STX mixture was intrarectally administered (both at 7.5 nmol/fish), and five individuals/group were analyzed after 1–48 h. Instrumental toxin analyses revealed that both TTX and STX were taken up, transferred, and retained, but more STX than TTX was retained in both groups. TTX gradually decreased and eventually became almost undetectable in the intestinal tissue, while STX was retained at ~5–10% of the dose level, and only STX showed transient transfer in the liver. The BW group showed a faster decrease/disappearance of TTX, greater STX retention in the intestine, and greater STX transient transfer to the liver. Thus, C. patoca appears to more easily accumulate STXs than TTXs, especially under hypoosmotic conditions.

The complete mitochondrial genome of the burrowing ghost shrimp Nihonotrypaea harmandi was reconstructed using the Illumina HiSeq platform. The genome was 15,272 bp in length made up of 37 mitochondrial genes (13 CDSs, 22 tRNAs, and 2 rRNAs) in the same order as other Nihonotrypaea species. Phylogenetic analyses suggested that Nihonotrypaea is a valid genus, and that N. harmandi can be phylogenetically marginally separated from N. japonica, though some authors considered the former as a synonym of the latter.