Molecular basis of the unique osmoregulatory strategy in the inshore hagfish, Eptatretus burgeri.

Hagfishes are characterized by omo- and iono-conforming nature similar to marine invertebrates. Conventionally, hagfishes had been recognized as the most primitive living vertebrate that retains plesiomorphic features. However, some of the "ancestral" features of hagfishes, such as rudimentary eyes and the lack of vertebrae, have been proven to be deceptive. Similarly, by the principle of maximum parsimony, the unique body fluid regulatory strategy of hagfishes seems to be apomorphic, since the lamprey, another cyclostome, adopts osmo- and iono-regulatory mechanisms as in jawed vertebrates. Although hagfishes are unequivocally important in discussing the origin and evolution of the vertebrate osmoregulatory system, the molecular basis for the body fluid homeostasis in hagfishes has been poorly understood. In the present study, we explored this matter in the inshore hagfish, Eptatretus burgeri, by analyzing the transcriptomes obtained from the gill, kidney, and muscle of the animals acclimated to distinct environmental salinities. Together with the measurement of parameters in the muscular fluid compartment, our data indicate that the hagfish possesses an ability to conduct free amino acid (FAA)-based osmoregulation at a cellular level, which is in coordination with the renal and branchial FAA absorption. We also revealed that the hagfish does possess the orthologs of the known osmoregulatory genes and that the transepithelial movement of inorganic ions in the hagfish gill and kidney is more complex than previously thought. These observations pose a challenge to the conventional view that the physiological features of hagfishes have been inherited from the last common ancestor of the extant vertebrates.

Long-term monitoring of egg-laying cycle using ultrasonography reveals the reproductive dynamics of circulating sex steroids in an oviparous catshark, Scyliorhinus torazame.

The many diverse reproductive strategies of elasmobranchs (sharks, skates and rays) from lecithotrophic oviparity to matrotrophic viviparity have attracted significant research attention. However, the endocrine control of elasmobranch reproduction is less well-documented largely due to their reproductive characteristics, such as a long reproductive cycle, and/or repeated internal fertilization using stored sperm in oviparous species. In the present study, for the first time, we succeeded in non-invasive monitoring of the continuing egg-laying cycle of the cloudy catshark Scyliorhinus torazame using portable ultrasound devices. Furthermore, long-term simultaneous monitoring of the egg-laying cycle and measurement of plasma sex steroids revealed cycling patterns of estradiol-17ß (E2), testosterone (T) and progesterone (P4). In particular, a decline in T followed by a reciprocal surge in plasma P4 were consistently observed prior to the appearance of the capsulated eggs, implying that P4 is likely associated with the ovulation and/or egg-case formation. While the cycling pattern of E2 was not as apparent as those of T and P4, threshold levels of E2 (>5 ng/mL) and T (>1 ng/mL) appeared to be crucial in the continuation of egg-laying cycle. The possibility to trace the dynamics of plasma sex steroids in a single individual throughout the reproductive cycles makes the catshark a useful model for regulatory and mechanistic studies of elasmobranch reproduction.

Measurement of 1α hydroxycorticosterone in the Japanese banded houndshark, Triakis scyllium, following exposure to a series of stressors.

An endocrine glucocorticoid response following exposure to a stressor has been well described for many vertebrates. However, despite demonstration of secondary stress responses in a number of elasmobranchs, our understanding of the endocrine control of these responses is lacking. This is largely due to the unusual structure of the dominant corticosteroid in elasmobranch fish, 1α-hydroxycorticosterone (1α-OH-B). Here we describe plasma extraction and HPLC separation procedures that allowed for the measurement of 1α-OH-B and corticosterone from plasma samples in the cannulated, conscious free-swimming Japanese banded houndshark, Triakis scyllium. While patterns of concentration in the plasma for 1α-OH-B and corticosterone were found to be similar in all experiments conducted, circulating levels of 1α-OH-B were consistently 100-fold greater than circulating levels of corticosterone. Immediately following cannulation surgery, circulating levels of 1α-OH-B increased 7-fold compared to pre-surgery levels, while the levels were 11-fold higher than pre-stress levels 30 min post a repeated handling/air-exposure stress. A three week period of fasting resulted in a 22-fold increase in circulating levels of 1α-OH-B in the banded houndshark. This is the first report of direct measurement of changes in circulating levels of the primary corticosteroid in elasmobranch fish, 1α-OH-B, following exposure to a stressor such as handling/air-exposure. Data indicate the steroid may respond similarly to the classic glucocorticoid response, such as cortisol in teleosts.

Evaluation of the aflatoxin biosynthetic genes for identification of the Aspergillus section Flavi.

Several fungi in the Aspergillus section Flavi have been widely used for fermentative food production, while some related species in the section are known to produce mycotoxin(s) that causes mycotic diseases. Common evolutionary markers, such as rRNA gene sequences and their internal transcribed spacers, cannot differentiate these non-aflatoxin-producing species from aflatoxin producers. Multilocus sequence analysis (MLSA) based on four aflatoxin biosynthetic genes encoding aflR, aflT, norA, and vbs, which are more variable nucleotide sequences than rRNA genes, can distinguish safe koji molds, A. oryzae and A. sojae, from aflatoxin-producing strains, A. flavus, A. toxicarius and A. parasiticus.