Theca layer and granulosa layer from six healthy Shanma duck at their peak production were collected. Real-time RT-PCR was used to detect the expression levels of AMH, AMHRⅡ, SF-1 and WT1. Our results showed that AMH was highly expressed in both SY(Small yellow follicle) and BW(Big white follicle), the expression level of AMH in theca layer was extremely significantly higher than that in the granulosa layer(P< 0.01); the expression pattern of AMHRⅡ in the granulosa layer was similar to that of AMH; SF-1 has a higher expression in the granulosa layer of preovulatoryfollicles, while showed a lower expression level in the theca layer. The expression levels of WT1 in theca layer of F1(The largest of preovulatory follicles), F3(Third largest preovulatory follicles), F6(the Smallest of preovulatory follicles) and SY follicular were extremely significantly higher than those in the granulosa layer(P< 0.01), while the expression levels in the granulosa layer of BW were the highest and extremely significantly higher than those in the BW theca(P< 0.01). In summary, the results suggested that AMH and its related genes were involved in the follicle development in duck, and showed various expression level in both granulosa and theca.

The aim of this study was to establish a Real-time quantitative PCR assay for detecting TLR4(Toll like receptor 4) gene in Sanhuang chicken, which provides a basis for the quantitative analysis of the expression level of TLR4 gene in Sanhuang chicken. According to the conserved region of the TLR4 gene of Sanhuang chicken, the primers were designed to establish Real-time PCR method using SYBR Green I dye. The standard curve was established by using the positive recombinant plasmid as the standard, and the dissolution curve was analyzed. The results showed that the established TLR4 gene Real-time PCR method had advantages of rapid high-throughput, wide linear range and high reproducibility. This study provided basis for further study on TLR4 gene of Sanhuang chicken.

Comb height was one of the important indirect selection index in breeding against sexual maturity, it was great significance to identify candidate genes that were highly correlated with comb height for quality chicken production. PCR-RFLP method had been utilized for genotyping in 1 301 individuals of NDH female populations, association analysis between the polymorphisms of the candidate genes GARNL1 with comb height was performed. The results showed that multiple loci of GARNL1 gene were significantly associated with comb height, and GARNL1 was a candidate gene for chicken comb height. Highly significantly associated(P<0.01) were found between rs14532831 and comb height at 77 d age, as well as rs15701119 was significantly associated with comb height at 77 d age(P<0.05); rs15701119, rs14532824 and rs14532831 could be used as effective molecular markers for comb height at 84 d age(P<0.01 or P<0.05); Moreover, 4 SNPs rs14532824, rs14532831, rs14532779 and rs15701119 were significantly associated with comb height at 91 d age(P<0.01 or P<0.05). The results demonstrated that GARNL1 gene had genetic effect on comb height.

Hypoxia inducible factor-1(HIF-1) is a transcription factor found in hypoxic cells, which plays a key role in the regulation of oxygen balance in vivo. Hypoxia inducing factor 1a(HIF-1α) is a key regulator of HIF-1 activation, which participates in the regulation of cell growth, differentiation and apoptosis. In order to further understand the hypoxia response system of Magang goose, this study designed primers by referring to the sequence of HIF-1α gene published on NCBI, and successfully cloned the HIF-1α gene mRNA, obtained the complete ORF sequence of HIF-1α, with a total length of 2 403 bp, which can encode 800 amino acids. The results showed that the structure of HIF-1α gene in Magang goose was conservative, and the homology of HIF-1α gene in poultry was high, and the highest homology with chicken was 94.1%. The protein structure showed that the main body contained helix loop helix, HLH, PAs(per Arnt SIM), PAC(motif C-terminal to PAS motifs) and other domains are consistent with the structural characteristics of HIF-1α protein. The results of tissue expression difference analysis show that HIF-1α gene is mainly expressed high in lung and pancreas, low in muscle and almost no expression in trachea.

Skeletal muscle development is strongly correlated with growth traits and establishing methods of goose myoblast isolation in vitro is vital for goose muscle development studies. Chest/leg muscle from Magang goose E15 was isolated by trypsin digestion and purified by differential attachment. Myogenic cells were identified by myogenic differentiation marker Myog and mature myofiber marker Myh1, together with immunofluorescence staining of myofiber marker MyHC. It was found that leg muscle oriented myogenic cells showed significantly higher proliferation and differentiation ability when compared to chest muscle, and myogenic cells under 39 ℃ culture showed higher proliferation and differentiation ability compared to that under 37 ℃. The method of goose embryonic myoblast isolation in vitro, which would benefit goose muscle development studies was successfully established.

Fibroblasts isolated from three well-developed 9, 11, 13 and 15 embryonic age embryos of Magang geese were cultured for 24, 48, 72, 96, 120 and 144 h, and the cell was counted, different age of embryo fibroblast growth curve was drawn, cell viability and cell proliferation rate was detected. The results showed that 9 and 11-day-old goose embryo fibroblasts were in better condition with fewer dead cells and hetero cells; the growth curve of 11-day-old goose embryo fibroblast cells was the best with the highest proportion of living cells; the proliferation rate of 9, 11 and 13-day-old geese embryo fibroblasts maintained higher levels. Eleven-days-old geese embryo was more suitable for isolation and culture of fibroblasts.

The period from embryonic stages to hatching day is the most critical period for goose muscle development, during which the development of myofiber and intramuscular fat play a vial role in goose muscle growth and meat quality traits. This study used chest and leg muscle of embryo 15, 23 d and hatching day from indigenous goose breeds-Shitou goose and Wuzong goose in Guangdong province. Through hematoxylin-eosin(HE), myosin ATPase and Oil red(ORO) staining, it was found that the leg muscle development of Shitou goose started earlier than that of Wuzong goose, but chest muscle of them had no significant difference; type Ⅰ myofibers were major myofiber type in leg muscle of two goose breeds, but the myofiber type was unclear in chest muscle; intramuscular fat in the chest and leg muscle of Shitou goose was later than that of Wuzong goose on embryo 23 d, while it was found significantly enriched than that of Wuzong goose on hatching day.

To investigate the effect of new large-scale water-saving breeding mode of breeding geese, taking two large-scale breeding goose farms in Guangdong Province as the research object, to detect the water quality changes of bath pond and water trough under different breeding modes of the two goose farms, and the effect of such changes on the productivity of breeding geese. The results showed that the total bacterial density, Escherichia coli density and endotoxin level in the bath water were significantly higher than those in the water trough under the semi-arid breeding model(P<0.05); the total bacterial density of open-pond breeding bath water was higher than that of semi-arid breeding bath water(P<0.01), but there was no significant difference in E.coli density(P>0.05); the endotoxin content in water bath in the open-pond breeding goose farm was significantly lower than that of the semi-arid goose farm(P<0.01), and the serum endotoxin level of the former was also low. Comparing the productivity of the two goose farms, it was found that the average daily laying rate in open-pond breeding goose farm was higher than that in semi-arid goose farm(P<0.01); while no significant difference was found in fertilizing rate, dead embryo rate and hatching rate between the two goose farms(P>0.05). The results showed that goose breeding enterprises could try to use semi-arid breeding model instead of the open-pond breeding.

In order to study the feasibility of DNA bar-coding technology in fish identification from the South China Sea, 65 species belonging to 43 genera, 28 families and 6 orders in the South China sea were collected and the COⅠ gene sequence of the species were amplified and determined, all sequences were aligned using BLAST from NCBI database for the species identification combining with the morphological information to identify and classify fishes. The sequence information, genetic distance and molecular phylogenetic tree of each species were calculated uing MEGA5.0 software. The results showed that the homologous sequence of COⅠ gene of 65 species were 633 bp. Comparing with the NCBI database, BLAST results with similarity rates of more than 99% can be obtained, which could be identified to species level. The average composition of A, T, G, C were 23.8%, 28.6%, 18.8% and 28.8%. The genetic distances among species, genera, families and orders were 14.88%, 19.09%, 23.14% and 24.68%, respectively. With the increasing of taxonomic category, the difference of average genetic distance became larger. In the phylogenetic tree, the species taxonomic relationships below family level were consistent with the morphological classification, while the relationships above family level were quite different from the morphological classification, indicated that the COⅠ gene is an effective bar code gene in fish molecular identification, but it is not necessarily suitable for the higher taxonomic level analysis for the molecular phylogenetic studies.

To investigate the phylogenetic relationships in the Lutjanus species, the partial sequences of 16 S rRNA and COⅠ genes of 12 Lutjanus species distributed in the South China Sea were amplified and sequenced. Combining the sequences of Lutjanus carponotatus, Lutjanus kasmira, Lutjanus gibbus and Lutjanus monostigma obtained from GenBank, the genetic distances were calculated, and the molecular phylogenetic trees were constructed using MEGA5.0 software. The results showed that the length of 16 S rRNA gene was 560 bp, and COⅠ gene was 639 bp. The sequence of 16 S rRNA gene was relatively conservative compared with COⅠ gene. The average inter-species genetic distances of 16 S rRNA gene and COⅠ gene were 0.039 and 0.146, respectively. Sixteen Lutjanus species were mainly divided into 4 branches in the molecular phylogenetic tree; moreover, the phylogenetic relationships in each branch showed a certain correlation with their morphological characteristics. There were several black spots on the upper back of Lutjanus russellii, Lutjanus fulviflamma and Lutjanus monostigma in the branch Ⅰ. In the branch Ⅱ, several longitudinal stripes can be observed in Lutjanus fulvus and Lutjanus kasmira, and some white spots exhibited can be observed on the upper back of Lutjanus rivalatus and Lutjanus stellatus. A large size was observed in Lutjanus sebae, Lutjanus erythropterus, Lutjanus malabaricus and Lutjanus gibbus in the branch IV; moreover the adult body colors of these fishes were bright red. In addition, Lutjanus species in branch Ⅰ exhibited a longer body size; however, the fishes showed a higher size in the branch Ⅳ. The body sizes in the branch Ⅱ and Ⅲ were between these observed in the branch Ⅰ and Ⅳ. Our results were consistent with previous studies, which suggested that the body size, coloration and band could be used as the basic classification characteristics of Lutjanus species.

To investigate the availability of using COⅠsequences as DNA barcodes to identify Gymnothorax species, partial sequences of mitochondrial cytochrome oxidase subunitⅠ(COⅠ) gene of 10 Gymnothorax speices from China sea were obtained by PCR amplification and sequencing. Combining with COⅠ gene sequence of 3 Gymnothorax species from GenBank, a total of 31 individual COⅠ sequences of 13 Gymnothorax species were obtained. Genetic distances of inter-and intra-species were calculated using MEGA 5.0 and the molecular phylogenetic trees were constructed using neighbour-joining and maximum likelihood methods. The results showed that intra-species genetic distance within the 13 Gymnothorax species were between 0.000-0.007 and the average genetic distance was 0.003; the inter-genetic distances among species were 0.005-0.243. Except for the distance between Gymnothorax pesudothyrsoideus and Gymnothorax reevesii was 0.005, inter-genetic distances among other Gymnothorax species were more than 2% and the average genetic distance was 0.187, indicating that the COⅠ gene could be used as the barcode gene for the Gymnothorax identification, but also had certain limitations. In the molecular phylogenetic tree, 13 Gymnothorax species mainly formed four branches, species Gymnothorax thyrsoideus was first separated and located at the base of the tree, indicating its lowest evolutionary status. Individuals of species G. pesudothyrsoideus and G. reevesii were crossed clustering together and could not formed each species monophyletic branch. However, there were significant differences in morphological character, excluding the possibility of synonyms, gene penetration caused by natural hybridization might exist between the two species. In addition, the COⅠ sequence of our sample Gymnothorax formosus was 5.5% different from the G. formosus obtained from GenBank, and their evolutionary status could also be clearly distinguished from the phylogenetic tree, indicating that species G. formosus sampled in this study may possibly be a new cryptic species.

Fucoidan is a water-soluble heteropolysaccharide mainly composed of L-fucose and sulfate groups, which has biological functions such as anti-oxidation, immune regulation, anti-inflammatory and anti-tumor, et al. In order to provide references for its application in the healthy breeding and safe production of animals, the physicochemical properties, extraction process, biological functions, as well as the actual application status in animal production were reviewed.