| Prussian carp(Carassius gibelio)is a gynogenetic fish(Carassius auratus gibelio ? × Cyprinus carpio ?),and because of its delicious meat and fast growth,it had become one of the important farmed fish and was extensively breeded throughout China.However,the outbreak of Cyprinid herpes virus ? infection in 2009,had caused huge economic loss of more than 50 million RMB and there had not been any effective solution up to now,which had very negative impact on the development of aquaculture of Prussian carp.In this paper,we analyzed the impact from environmental bacterial communities and Cyprinid herpes virus ? infection on intestinal bacterial community of Prussian carp by high-throughput sequencing and metabolomics to find out the correlation between Cyprinid herpes virus ? infection and enviromental bacterial communities.38 Water and sediment samples were collected seperately from 5 ponds in Da feng,Jiangsu province in May and June,2015 for normal bacterial community study of aquaculture water and sediment;135 gut samples of healthy fish from the same 5 ponds on the same time points were collected for normal intestinal bacterial community research;92 gut samples of healthy and diseased fish from another two ponds were collected to study the impact of Cyprinid herpes virus ? infection on intestinal bacterial community of Prussian carp.Results showed that: 1.Variation analysis of bacterial communities in normal aquaculture waters from different ponds and different time pointsBacterial communities in normal aquaculture waters from different ponds and different time points were analyzed by high-throughput sequencingand statistic analysis.Results showed that:(1)Variation fromdifferent time points: there was no significant difference in ?-diversity of bacterial community between the two different time points.In the 8 dominant phylum,there was 2 dominant phylum with abundances declined significantly in June;In the 7 dominant genuses,there were 4 dominant genuses with abundances declined significantly in June.(2)Variation fromdifferent ponds: there was significant difference in ?-diversity of bacterial community among the 5 ponds.Significant differences were also found in the abundances of 7 out of the 8 dominant phylum and all of the 7 dominant genuses amongdifferent ponds.2.Variation analysis of bacterial communities in normal sediments from different ponds and different time pointsBacterial communities in normal sediments from different ponds and different time points were analyzed by high-throughput sequencingand statistic analysis.Results showed that:(1)Variation fromdifferent time points:there was no significant difference in ?-diversity of bacterial community between the two different time points.In the 7 dominant phylum,there was 1 dominant phylum with abundance declined significantly and 2 dominant phylum with abundances rose significantly in June.In the 13 dominant genuses,there was 1 dominant genus with abundance declined significantly and 1 dominant genus with abundance rose significantly in June.(2)Variation fromdifferent ponds: there was significant difference in ?-diversity of bacterial community among the 5 ponds.Significant differences were also found in the abundances of 3 out of the 7 dominant phylum and 11 out of the 13 dominant genuses amongdifferent ponds.3.Variation analysis of intestinal bacterial communities in normal Prussian carp from different ponds and different time pointsIntestinal bacterial communities in normal Prussian carp from different ponds and different time points were analyzed by high-throughput sequencingand statistic analysis.Results showed that:(1)Variation fromdifferent time points:?-diversity of intestinal bacterial community of Prussian carp was declined significantly in June.In the 10 dominant phylum,there was 6 dominant phylum with abundances declined and 1 dominant phylum with abundance rose in June.In the 8 dominant genuses,there were 4 dominant genuses with abundances declined and 3 dominant genuses with abundances rose in June.(2)Variation fromdifferent ponds: there was significant difference in ?-diversity of bacterial community among the 5 ponds.Significant differences were also found in the abundances of all of the 10 dominant phylum and the 8 dominant genuses amongdifferent ponds.4.Variation analysis between environmental and intestinal bacterial communities of Prussian carpDifference among bacterial communities of aquatic water,sediment and intestine of Prussian carp were analyzed,results showed that: Bacterial community diversity of sediment samples was higher than that of water samples,bacterial community diversity of water samples was higher than that ofintestinal bacterial communities.On phylum level,there was 7 same dominant phylum between the bacterial community of gut and water samples;there was 6 same dominant phylum between the bacterial community of gut and sediment samples;there was 6 same dominant phylum between the bacterial community of sediment and water samples.On genus level,there were 2 same dominant genuses in the bacterial community of gut and water samples;there was' t same dominant genus between the bacterial community of gut and sediment or water and sediment samples.On phylum level,the abundances of Fusobacteria in gut and water samples rose significantly in June,but did not change in sediment;the abundances of Actinobacteria in water and sediment samples rose significantly in June,but declined significantly in gut samples.The abundances of Bacteroidetes in water and sediment samples declinded significantly in June,but rose significantly in gut samples.On genus level,the abundances of Synechococcus in sediment samples declined significantly in June,but rose significantly in gut and water samples.The abundance of Anoxybacillus rose significantly in sediment samples,but declined significantly in gue and water samples.5.Impact of Cyprinid herpes virus ? infection on intestinal bacterial microbiota of Prussian carpIntestinal bacterial communities in healthy and diseased Prussian carp were analyzed by high-throughput sequencingand statistic analysis.Results showed that: there was significant difference in ?-diversity of bacterial community between healthy and diseased fish.In the 11 dominant phylum,there was 6 dominant phylum with abundances declined and 3 dominant phylum with abundances rose in diseased fish.In the 11 dominant genuses,there were 4 dominant genuses with abundances declined and 4 dominant genuses with abundances rose in diseased fish.Plesiomonas could be used as a baterial biomarker for CyHV-2 infection.6.Impact of Cyprinid herpes virus ? infection on intestinal metabolites of Prussian carpIntestinal metabolites in healthy and diseased Prussian carp were analyzed by metabolomics and statistic analysis.Results showed that: intestinal metabolites of Prussian carp were dominated by amino acid,fatty acid,choline,purine,et al.Intestinal metabolites were changed obviously after CyHV-2 infection: Underpositive scan mode,abundancess of 5073 metabolites rose and 2850 metabolites declined;under negative scan mode,abundances of 1597 metabolites rose and 1556 metabolites declined.Compared with online database,20 metabolites with abundances declined and 4 metabolites with abundances increased were identified.The abundances of Immunoenhancers such as n-Octyl-beta-D-thioglucopyranoside,Daidzein,Genistein declined significantly.In conclusion,by high-throughput sequencing and metabolomics,impact from environmental bacterail communities and Cyprinid Herpesvirus 2(CyHV-2)infection on intestinal bacterial community of Prussian carp was studied.Results showed that: intestinal microbiota of Prussian carp was influenced by environmental bacterial communities and CyHV-2 infection.Plesiomonas can be used as bacterial biomarker for CyHV-2 infection.Through metabolomics analysis,CyHV-2 infection changed intestinal metabolomics of Prussia carp and a variety of immunoenhancerswere found to decline after CyHV-2 infection.All these results can provide basic reference for disease prevention and treatment in aquaculture of Prussian carp. |
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