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Most canopy-forming macroalgae have disappeared from temperate reefs in southern Japan, one of the ocean warming hotspots, but Sargassum nipponicum is surviving in this region. As this species’ annual shoots emerge from holdfasts during summer, both plant components may be highly tolerant to warm and nutrient-poor conditions in this season. The present study examined the effects of temperature and nutrient conditions on holdfast growth, shoot emergence from holdfasts, and shoot growth in S. nipponicum samples collected in Tanegashima Island, southern Japan. The summer temperature in this region (30 °C) allowed holdfast growth and shoot emergence but inhibited shoot growth. Nutrient-poor conditions had limited effects on the first two parameters but suppressed shoot growth. These results suggested that during warm summers and under nutrient-poor conditions in southern Japan, shoots can emerge from S. nipponicum holdfasts but cannot further grow. Additionally, nutrient loading from a nearby river was higher at the only site dominated by S. nipponicum, than at the other sites where this species was absent on Tanegashima Island. This was observed especially between autumn and winter, implying that such a nutrient-rich environment may contribute to shoot growth in S. nipponicum and to the persistence of its population in the area.

The quantification of histidine-containing dipeptides (anserine, carnosine, and balenine) in serum might be a diagnostic tool to assess the health condition of animals. In this study, an existing reversed-phase ion-pair high-performance liquid chromatography (HPLC)–ultraviolet detection method was improved and validated to quantify serum anserine, carnosine, and balenine levels in the dolphin. The serum was deproteinized with trichloroacetic acid and directly injected into the HPLC system. Chromatographic separation of the three histidine-containing dipeptides was achieved on a TSK–gel ODS-80Ts (4.6 mm × 150 mm, 5 µm) analytical column using a mobile phase of 50 mmol/L potassium dihydrogen phosphate (pH 3.4) containing 6 mmol/L 1-heptanesulfonic acid and acetonitrile (96:4). The standard curve ranged from 0.1 µmol/L to 250 µmol/L. The average accuracy of the intra- and inter-analysis of anserine, carnosine, and balenine was 97–106%. The relative standard deviations of total precision (RSDr) of anserine, carnosine, and balenine in dolphin serum were 5.9%, 4.1%, and 2.6%, respectively. The lower limit of quantification of these compounds was 0.11–0.21 µmol/L. These results indicate that the improved method is reliable and concise for the simultaneous determination of anserine, carnosine, and balenine in dolphin serum, and may be useful for evaluation of health conditions in dolphins. Furthermore, this method can also be applied to other biological samples.