Study On Genetic Diversity And Skin Adjunct’s Characteristics Of Chinese Giant Salamander

The Chinese Salamander Andrias davidianus is a specific endangered animal as the second-class protected species. We had established16giant salamander protected areas and some nature reserves (giant salamander as a protected directory). The research on the suitable environment and artificial propagation of the giant salamander is conducted. It has gradually established the artificial breeding system, which can protect the species effectively.There are some deficiencies in current protection of giant salamander:(1) The confusion of germ plasm resources and inbreeding leads to the degradation of giant salamander, which is not conducive to long-term survival of the species. So it’s necessary to analyze genetic diversity and phylogenetic relationships among the parents of the artificial breeding of Giant Salamander to provide reasonable protection strategy.(2)There’s lack of sustainable protection approach. It is a natural physiological phenomenon that giant salamander produces the skin appendages such as shedding and secretion many times in life, belonging to the renewable materials for rational development and utilization. The rational use of these is more suitable for the long-term protection of giant salamander resources. This study establishes a non-injury sampling technique to extract genomic DNA using the skin appendages like sheding and secretion of giant salamander. And then we screen marker-site of SSR and analyze the genetic diversities and phylogenetic relationships of giant salamander parents, which provide the basis for the long-term protection strategy of the giant salamander. It provides us the support for the full and rational use of the giant salamander skin appendages with the analysis of their composition and physicochemical properties, combing the animal model to evaluate the specific effect:(1) The genetic diversity analysis:Skin secretion and shedding is a good source for non-destructive genetic sampling in the giant salamande. The amplification of the12S sequences confirmed that the DNA obtained can be used for the analysis of genetic structure.34microsatellite sequences was amplified from tissue samples of genomic DNA, and10microsatellite sequence with polymorphism which can be used in the analysis of genetic diversity of giant salamander resources. The artificial breeding of giant salamander parents in Zhangjiajie giant salamander core protected areas have a wealth of genetic diversity, whose cluster analysis shows it’s fall into four populations.(2) Physical and chemical properties of the secretion and activity analysis:highly hydrated gel material, the main component is protein, soluble in only5%(V/V) acetic acid and protease solution. It has a strong viscosity; average molecular weight is3.61×106. The secretions of the soluble fraction have the protease hydrolysis activity and phospholipase A2activity. It does not have significant antibacterial activity and only weak inhibition of microbial growth activity at an early stage.The oxidation resistance characteristics clearly significant effect on DPPH and ABTS scavenging activity,90.83%and93.12%at0.6mg/ml. The peptides by thermal denaturation at0.05mg/ml are the ABTS scavenging activity with70.90%.(3) The anti-tumor activity of the secretion:The giant salamander skin secretions of soluble fraction have lethal activity on mouse; LD50(intraperitoneal) value is32.95mg/kg. The active group is heat-labile protein. Anatomy of the death individual found that lung tissue changes significantly and have the phenomenon of congestion, convulsions which may cause the death of mouse. The secretion produce local effects on animals including the activity of significant pain, increase vascular permeability, persistent swelling, allow muscle tissue swelling and necrosis, but no significant bleeding activity. In vivo, it can Inhibit the growth of ascites tumor cells, reduce the number of ascites tumor cells and prolong survival, it could significantly improve blood parameters in mice, did not bring about significant changes in liver structure. In vitro, inhibiting the growth of both cancer cells and normal cells are different in different cells, and the mechanism is not to promote the apoptosis of tumor cells.(4) The giant salamander shedding composition and treatment on burns:giant salamander shedding contains rich amino acids, mineral elements, and eight kinds of fatty acids. Large dose of the giant salamander shedding ointment can be significantly improved wound healing rate, shortened wound healing time and accelerate wound healing. The inhibition of inflammation cytokines may be one of the mechanisms.