Gel Microbead Cultivation with a Subenrichment Procedure Can Yield Better Bacterial Cultivability from a Seawater Sample than Standard Plating Method

A gel microbead (GMD) cultivation method was employed to cultivate microorganisms from an amphioxus breeding zone in Qingdao, P. R. China. The culture results were compared with those by standard plating method. In the GMD-based method, the microcolony-forming GMDs were sorted by fluorescence-activated cell sorting (FACS). To further get pure cultures, a subsequent enrichment culture and a streaking purification procedure were conducted on marine R2A medium. Eighty bacterial strains isolated by the GMD-based method were randomly selected for sequencing. These isolates belonged to Alphaproteobacteria (33%), Gammaproteobacteria (44%), Bacteroidetes (11%), Actinobacteria (5%), Firmicutes (5%), Epsilonproteobacteria (1%), and Verrucomicrobia (1%), the last two groups being usually difficult to culture. The 16S rRNA gene sequences revealed a diverse community with 91.1%-100% of the bacterial rRNAs similarities. Thirteen strains were sharing 16S rRNA gene sequence which was less than 97% similar to any other rRNA genes currently deposited in TYP16S database. Seventy isolates derived from the standard plating method fell into 4 different taxonomic groups: Alphaproteobacteria (9%), Gammaproteobacteria (81%), Bacteroidetes (7%) and Firmicutes (3%) with a 16S rRNA gene sequence similarities between 95.8%-100%, in which only 3 strains were sharing 16S rRNA gene sequence of less than 97%. The results indicated that the GMD-based method with subenrichment culture yielded more taxonomic groups and more novel microbial strains, including members of previously rarely cultured groups, when compared with the standard plating method, and that this method markedly improved the bacterial cultivability.

Accelerating column leaching trial on copper sulfide ore

The main measures to accelerate leaching sulfide ore are large spraying intensity,manual oxygen supply,temperature control,and acclimated bacteria.The indoor experiment accelerating sulfide ore leaching detected the temperature during leaching process,dissolvability of oxygen,bacterial concentration,Cu concentration and slag grade.At the same time,this paper also analyzed the effect of four factors,which are bacterial diversity cultivation stage,spraying intensity,air supply,and whether to control temperature,on the leaching efficiency of copper.The results indicate that the oxygen content of leach solution has a close relationship with temperature but it is rarely affected by air supply.The bacterial concentration preserves from 106 to 107 mL^-1,and temperature has a great effect on the bacterial activity under the condition of proper temperature and oxygen supply,and the lack of nutrition prevents the bacterial concentration from rising in the late stage.The relationships of the copper leaching efficiency to temperature,air feed,and spraying intensity are directly proportional.The leaching efficiencies of the cultivated bacteria and acclimation bacteria are 1.2 and 1.4 times as large as that of the original bacteria.