The early embryonic development of the green crucian carp ( Carassius auratus indigentiaus subsp. nov.) was observed to study its timing and characteristics. The fertilized eggs are round, slightly yellow, and viscous demersal. The egg diameter after water swelling was 1.47 ± 0.04 mm. Embryonic development can be divided into eight stages according to its major characteristics: blastoderm formation, mitotic, blastula, gastrula, neurula, blastopore closure, organogenetic and hatching stages. Under a water temperature of 24 ± 1 °C, salinity of 35 ± 1, and pH of 7.4 ± 0.5, the blastoderm began to form 35 min after fertilization. It entered the mitotic stage at 55 min, blastula stage at 220 min, gastrula stage at 460 min, neurula stage at 675 min, blastopore closure stage at 700 min, organogenetic stage at 900 min and hatching stage at 3390 min. The total length of newly hatched larvae was 4.07 ± 0.35 mm. Regression models of growth characteristics were obtained. The full-length growth rate was fastest from 15 to 26 days, with an average of 0.396 mm/day. Compared with other cyprinid fishes, green crucian carp exhibited some distinct characteristics in certain stages of embryonic development. The eye primordium developed before the sarcomere, and the heart rate was relatively high before the member stage. Yolk fluctuation was observed during the multi-cell phase of embryonic development. The sarcomere formed after the eye primordium. The heart rate in the hatching phase was 136 beats/min. This study provides a reference for embryonic development in green crucian carp, which will assist its large-scale cultivation.

The estrogen receptor signaling pathway plays an important role in vertebrate embryonic development and sexual differentiation. There are four major estrogen receptors in zebrafish: esr1, esr2a, esr2b and gper. However, the specific role of different estrogen receptors in zebrafish is not clear. To investigate the role of esr2b in zebrafish development and reproduction, this study utilized TALENs technology to generate an esr2b knockout homozygous zebrafish line. The number of eggs laid by esr2b knockout female zebrafish did not differ significantly from that of wild zebrafish. The embryonic development process of wild-type and esr2b knockout zebrafish was observed, revealing a significant developmental delay in the esr2b knockout zebrafish. Additionally, mortality rates were significantly higher in esr2b knockout zebrafish than in their wild-type counterparts at 24 hpf. The reciprocal cross experiment between esr2b knockout zebrafish and wild-type zebrafish revealed that the absence of esr2b resulted in a decline in the quality of zebrafish oocytes, while having no impact on sperm cells. The knockout of esr2b also led to an abnormal sex ratio in the adult zebrafish population, with a female-to-male ratio of approximately 1:7. The quantitative PCR (qPCR) and in situ hybridization results demonstrated a significant downregulation of cyp19ab1b expression in esr2b knockout embryos compared to wild-type embryos throughout development (at 2 dpf, 3 dpf and 4 dpf). Additionally, the estrogen-mediated induction expression of cyp19ab1b was attenuated, while the estradiol-induced upregulated expression of vtg1 was disrupted. These results suggest that esr2b is involved in regulating zebrafish oocyte development and sex differentiation.

Chinese herbal medicine is rich in active ingredients that can promote growth and enhance immune function. In this study, Lycium barbarum, Panax ginseng, Astragalus membranaceus and Phragmitis rhizoma were crushed and mixed to prepare a compound Chinese herbal medicine. The basic feed of Megalobrama amblycephala was supplemented with 0 (control group), 1% (T1), 2% (T2) and 4% (T3) of this compound medicine. After raising for 90 days, in the T1 and T2 experimental groups, the length and width of intestinal villi and the activities of amylase, trypsin and lipase in the intestine were significantly higher than those in the control group. The weight gain rate and specific growth rates were highest and the feed coefficient was lowest in the T2 experimental group. In the control group, a large number of dilated hepatic sinusoids were detected, while this number significantly decreased in the T1 experimental group and they were not detected at all in the T2 and T3 experimental groups. The spleen and liver body indices were highest in the T2 experimental group. In all experimental groups, the Lys content and the activities of T-SOD, CAT, ACP, AKP and GSH-PX in serum were significantly higher than those of the control group. The expression of IgM, C3, TNF-ɑ and IL-1β in the head kidney; C3, TNF-ɑ and IL-1β in the spleen; C3 and IL-1β in the gills; IgM, C3 and IL-1β in liver; and IL-1β in the intestine was highest in the T2 experimental group. After challenge with Aeromonas hydrophila, the cumulative mortality rate of M. amblycephala was lowest in the T2 experimental group. The results of this study indicated that this compound Chinese herbal medicine could significantly enhance immunity, increase the activity of intestinal digestion-related enzymes and promote the growth of M. amblycephala. The appropriate addition amount of this compound Chinese herbal medicine in the basic feed of M. amblycephala was 2%.