Research On The Effects Of Yong Cheng Gangue Pollution On Carassius Auratus Muscle And Spleen

Objective： Heavy metal content of soil and water samples were detected in Yongcheng contaminatedareas of coal gangue, and selecting carp samples to study the index changes of spleen and muscle. Thepurpose was to investigate the effects of gangue pollution on aquatic organisms, and provide experimentalevidence for monitoring and controlling the coal gangue contamination.Methods： The samples of soil, water and fish, which were collected from Yongcheng coalcontaminated areas, acted as the experimental group, and Tianmu Lake District was control group. Soil andwater pH was measured at room temperature using a pH meter, collecting soil and water samples weredigested by Automatic Graphite Digestion System, the obtained test preparations were used to detect heavymetals content (Cu, Cd, Pb, Cr, Zn) by atomic absorption spectrometry, combining with The SoilEnvironment Degree of Quality Standard (GB15618-1995) and Surface Water Quality Standard(GB3838-2002) to evaluate heavy metal pollution.Both groups of ten crucian carp samples wereanatomized and isolated the spleen and muscle tissue, then we obtained tissue homogenate and meauredNa+K+-ATPase, Superoxide Dismutase(SOD), Catalase(CAT), Glutathione Peroxidase(GSH-Px)activity;Separation of the spleen and muscle cells can use the enzyme digestion method, with the Single CellGelelectrophoresis (SCGE) discussed and analyzed whether the DNA of tissue cells were damaged.Spleenand muscle tissue with a neutral formaldehyde fixed, the conventional dehydration, embedding madeparaffin section, using histological staining party (Haemato-xylin and Eosin, HE), to observe the spleen andmuscle morphological changes. There are three methods such as drip loss method, freeze drying andcentrifugation for detection of the cpacity of muscle tissue water-holding changes; Muscle samples aftervacuum freeze drying, dry, with a constant element analyzer (CNS) determination of contentof C,N and S. Results：①The detection to heavy metal content in soil and gangue contaminated water：Bothcontaminated soil and water pH were slightly acidic. Determination of heavy metals showed that soil Cu,Cd, Pb, Cr, Zn contents were757.38mg/kg,1.31mg/kg,762.54mg/kg,502.23mg/kg,753.87mg/kg,higher than the11.61mg/kg,0.15mg/kg,23.28mg/kg,13.51mg/kg,27.84mg/kg, five heavy metals hadbeen more than the reference amount of soil environmental quality standards (GB15618-1995), in whichCu, Cd, Pb polluted farther. The content of Cu, Cd, Pb, Cr, Zn in water was1.33mg/L,0.03mg/L,0.24mg/L,0.39mg/L,2.58mg/L, higher than the0.37mg/L,0.01mg/L,0.05mg/L,0.04mg/L,0.81mg/L,which Cu, Pb, Cr content exceeded Surface Water Quality Standard (GB3838-2002) reference values andthe water were moderately polluted.②Impact of pollution on fish gangue spleen：it can be seen from theenzyme activity assay, ten samples of contaminated fish, the ATPase and GSH-Px activity of spleencompared with the control group were significantly lower (P<0.05), SOD and CAT activity decreasereached extremely significant level (P<0.01). According to the single cell gel electrophoresis test results,the carp spleen cells of experimental group were mixed with varying degrees of tailing and DNA chainbroke, while the control group did not have this phenomenon, CASP analysis showed that the total lengthof the comet (CL), tail length (TL), tail moment (TM) and Olive tail moment (OTM) are increased, which,TL and TM parameters were increased5.3μm,1.39μm and reached a significant level (P<0.05). HEstaining showed that organizations had more vacuoles, spleen blood sinus cavity became larger, red pulpand white pulp were not easy to distinguish, the regions of reticular cells and lymphocyte appeared necrosis,there were a large number of nodules stained by HE.③Coal pollution impact on fish muscle：muscletissue SOD, CAT, were significantly lower ATPase, GSH-Px activity compared with the control group (P0.05). It can be seen from single cell gel electrophoresis images, the mixed muscle cells of experimentalgroup carp, compared with the control group, happened to tail and DNA without breakage. CASP analysis showed that the comet total length (CL), tail length (TL), tail moment (TM) and Olive tail moment (OTM)did not change significantly (P>0.05). The method to drip loss, lyophilization, centrifugation weredeterminated muscle loss rate, and results showed the test group was significantly lower than the controlgroup (P0.05), the cooked meat was also significantly lower than the control group (P0.05). The testgroup N, C percentages of Muscle tissue are lower than in the control group, while the S content wassignificantly higher (P0.05), the C/N (mol ratio) in the experimental group is slightly higher than thecontrol group.Conclusions：①Coal contaminated soil and water were tested slightly acidic, the content of soilheavy metals Cu, Cd, Pb, Cr, Zn had been more than "Soil Environmental Quality Standard(GB15618-1995)", Order of degree of pollution was Cu> Cd> Pb> Cr> Zn, which, Cu, Cd, Pb pollution isheavier. Content of contaminated water Cu, Pb, Cr exceeded "Surface Water Environmental QualityStandard (GB3838-2002)", excessive pollution is moderate, suggesting that heavy metal pollution causedby the coal gangue and severely damaged the ecological balance of the surrounding soil and water.②Heavy metal pollution made carp spleen, muscle ATPase, SOD, CAT, GSH-Px activity significantlyreduced,and antioxidant defense system had been severely destroyed; spleen cell DNA damage was heavy,there was no significant change in muscle cell DNA damage; spleen organization structure changed, muscletissue had no pathological features spleen.③Heavy metal pollution made the water holding capacity tocarp muscle significantly reduced; C and N content reduced but S content increased, which led to thesynthesis of protein and fat slow. Dressing percentage of fish is also lower.