Research On Characteristics Of Bacterial Community In Ballast Water Of Typical Routes In Shanghai

With the development of the world economy and the globalization of trade,maritime transportation has accounted for more than 80% of global trade and transportation.Species invasion caused by ship's ballast water has become a hot spot of concern.The ship's ballast water is used to control its trim,heel,draught and stabilize the water and suspended solids contained in the ballast tank during the ship's navigation.The research on microbial in ship ballast water is a research hotspot of port intrusion ecology and ship ballast water management and control.This paper investigates the ballast water bacteria in the ship by selecting 32 ships that are docked on three typical routes?China-America route,China-Europe route,China-Australia route?on Shanghai Yangshan Port.Enrichment culture,combined with molecular biology method 16 SrRNA first generation sequencing and second generation high-throughput sequencing technology,studied the diversity of bacteria in ship ballast water,and studied the phylogenetic relationship of culturable bacteria.The bacterial composition of the ship's ballast water varies in different routes.In this study,the 16 SrRNA database of the ballast water bacteria in Shanghai Yangshan Port was enriched by the investigation of the bacterial community composition of different routes of ship ballast water,which provided the basic data of molecular biology for understanding the bacterial diversity of ship ballast water.The alien species invasive ecology provides theoretical basis and support.The specific research results are as follows: 1.A total of 16 samples of ship ballast water were collected through sample collection of 14 vessels on the China-America route.The ballast areas are Pacific Ocean,Atlantic Ocean,East China Sea,China Yellow Sea,Philippine Sea,Colombo Port,etc.The sample distribution includes four seasons of spring,summer,autumn and winter.The ballast water age ranges from 3 to 145 days,including the ship COSCO ISTANBUL water.The age of the ship is 145 days,the ship EVEN UNIQUE is 103 days old,and the other 14 ships are under 68 days.The total number of bacterial OTUs is 3799,and the number of OTUs between different ships ranges from 167 to 1069,with a wide range of differences.High-throughput sequencing analysis included 41,93,209,401,870,and 1625.The dominant group is ?-proteobacteria?Gammaproteobacteria 45.97%?,?-proteobacteria?Alphaproteobacteria 25.24%?,Flavobacteriia?6.27%?,Actinobacteria?3.68%?,?-Proteobacteria?Betaproteobacteria 3.29%?.The culturable bacteria were isolated from HG,HP,MA and YM,and 26 strains of bacteria were isolated and their sequences were compared with 16 SrRNA genes,belonging to 3 gates,5 classes,8 orders,13 families.,14 genera,24 species.At the gate level,the dominant group is Proteobacteria?84.6%?and Bacteroidetes?11%?.At the genus level,the dominant genus is Pseudoalteromonas.2.A total of 15 samples of ship ballast water were collected through sample collection of 14 vessels on the China-Europe route.The ballast sites are the East China Sea,the South China Sea,the Yellow Sea in China,the high seas near Taipei,the Indian Ocean,etc.The sample distribution includes the four seasons of spring,summer,autumn and winter.The ballast water age ranges from 0 to 439 days,of which the ship COSCO The water age of FRANCE is 439 days,and the remaining 13 ships are all in the range of 0 to 55 days.The total number of bacterial OTUs is 3,396,and the number of OTUs between different ships ranges from 238 to 923.High-throughput sequencing analysis included 39,86,197,380,817,and 1495.The dominant group is ?-proteobacteria?Gammaproteobacteria 36.09%?,?-proteobacteria?Alphaproteobacteria 30.15%?,Flavobacteriia?8.94%?,and ?-Proteobacteria?7.23%?.The CM,CS,EM,EV,and ME were isolated for culturable bacteria.A total of 39 strains of bacteria were isolated and their sequences were sequenced with 16 SrRNA genes,belonging to 4 gates,6 classes,12 orders,13 Division,17 genera,30 species.The dominant group is Proteobacteria?69%?and Bacteroidetes?23%?.At the genus level,the dominant genus is Erythrobacter.3.Through the sample collection of 4 ships on the China-Australia route,a total of 4 ship ballast water samples were collected.The ballast sites are in the Pacific Ocean and Osaka Port.The sample distribution includes the spring,summer and autumn seasons.The ballast water age ranges from 35 to 156 days.The total number of bacterial OTUs is 1238,and the number of OTUs between different ships ranges from 355 to 523,with a small difference.High-throughput sequencing analysis included 29,67,144,258,461,and 692.?-Proteobacteria?Gammaproteobacteria 48.0%?,?-Proteobacteria?Alphaproteobacteria 19.66%?,Pltb_vmat₈0 12.39%,norank_pMarinimicrobia_SAR406_clade 4.80%.The GM and NF of the ship were separated by culturable bacteria,and 12 strains of bacteria were isolated and their sequences were compared with 16 SrRNA genes,belonging to 2 gates,3 classes,6 orders,8 families and 9 genera.,12 species.The dominant group is Gammaproteobacteria and Alphaproteobacteria,which account for 50% and 42%,respectively.At the genus level,the dominant genus is Erythrobacter.