Vitro culture of axe-head glochidia in pink heelsplitter Potamilus alatus and mechanism of its high host specialists.

The basal media M199 or MEM was utilized in the classical method of vitro culture of glochidia where 1-5% CO2 was required to maintain stable physiological pH for completion of non-parasitic metamorphosis. The classical method encounters a great challenge to those glochidia which undergo development of visceral tissue but significantly increase in size during metamorphosis. The improved in vitro culture techniques and classical methods were firstly compared for non-parasitic metamorphosis and development of glochidia in pink heelsplitter. Based on the improved method, the optimal vitro culture media was further selected from 14 plasmas or sera, realizing the non-parasitic metamorphosis of axe-head glochidia for the first time. The results showed that addition of different plasma (serum) had significant effect on glochidial metamorphosis in pink heelsplitter. Only glochidia in the skewband grunt and red drum groups could complete metamorphosis, the metamorphosis rate in skewband grunt was 93.3±3.1% at 24±0.5°C, significantly higher than in marine and desalinated red drum. Heat-inactivated treatment on the plasma of yellow catfish and Barbus capito had significant effect on glochidia survival and shell growth. The metamorphosis rate also varied among different gravid period, and generally decreased with gravid time. Further comparison of free amino acid and fatty acid indicated that the taurine of high concentration was the only amino acid that might promote the rapid growth of glochidial shell, and the lack of adequate DPA and DHA might be an important reason leading to the abnormal foot and visceral development. Combined with our results of artificial selection of host fish, we tentatively established the mechanism of its host specialists in pink heelsplitter for the first time. This is the first report on non-parasite metamorphosis of axe-head glochidia based on our improved vitro culture method, which should provide important reference to fundamental theory research of glochidia metamorphosis and also benefit for better understand of mechanism of host specialists and generalists of Unionidae species.

Monogonont Rotifer, Brachionus calyciflorus, Possesses Exceptionally Large, Fragmented Mitogenome.

In contrast to the highly conserved mitogenomic structure and organisation in most animals (including rotifers), the two previously sequenced monogonont rotifer mitogenomes were fragmented into two chromosomes similar in size, each of which possessed one major non-coding region (mNCR) of about 4-5 Kbp. To further explore this phenomenon, we have sequenced and analysed the mitogenome of one of the most studied monogonont rotifers, Brachionus calyciflorus. It is also composed of two circular chromosomes, but the chromosome-I is extremely large (27 535 bp; 3 mNCRs), whereas the chromosome-II is relatively small (9 833 bp; 1 mNCR). With the total size of 37 368 bp, it is one of the largest metazoan mitogenomes ever reported. In comparison to other monogononts, gene distribution between the two chromosomes and gene order are different and the number of mNCRs is doubled. Atp8 was not found (common in rotifers), and Cytb was present in two copies (the first report in rotifers). A high number (99) of SNPs indicates fast evolution of the Cytb-1 copy. The four mNCRs (5.3-5.5 Kb) were relatively similar. Publication of this sequence shall contribute to the understanding of the evolutionary history of the unique mitogenomic organisation in this group of rotifers.

[Effects of diets on growth, serum biochemical indices and lipid metabolism in Coilia nasus].

Effects of diets on growth, serum biochemical indices, and enzyme activities related to lipid metabolism in fingerlings Coilia nasus at age of 6 months were investigated during 60-day experiment in this study. Fingerlings with similar body length and mass were fed with one of 3 types of diets (diet 1: soft pellet; diet 2: soft pellet mixed with fish oil; diet 3: slow-sinking hard pellet). Fish fed with diets 2 or 3 had significantly higher total body mass, rate of mass gain, specific growth rate, and fullness coefficient than those fed with diet 1. Fish fed with diet 3 exhibited the lower food coefficient compared to those fed with diets 1 or 2. Growth traits (length, length to mass ratio, length to width ratio, hepatopancreas somatic indices and viscera somatic index) and serum biochemical indices (total protein, albumin, blood glucose, cholesterol and triglycerides) in all three treatments were not significantly different. Fish fed with diet 1 exhibited significantly higher carnitine palmitoyltransferase-I than those fed with diets 2 or 3, while fish fed with diet 2 exhibited significantly lower carnitine palmitoyltransferase-II. However, amylase, pepsin, lipase activity, lipoprotein lipase and acetyl-coa carboxylase had no significant difference in fish body among all groups. Results suggested that fish oil as a diet supplement highly facilitated fish growing. The slow-sinking pellet had the highest utilization efficiency and was suitable to feed fish fingerlings of C. nasus.