养殖池塘底泥预处理方法对Biolog Eco板分析结果的影响

Effects of Preprocessing Methods of Sediment on Analysis Results of Biolog EcoPlates

为探讨不同预处理方法对养殖池塘底泥Biolog Eco板分析结果的影响,以南美白对虾养殖池塘周边底泥1#和中部底泥2#为研究对象,考察了文献中常用的5种预处理方法底泥微生物群落平均颜色变化率( Average well color development,AWCD)、碳源利用模式和多样性指数的差异。结果表明,培养3 d后周边底泥1#样品异养细菌总数(7.73×10^5 CFU·mL^-1)显著低于中部底泥2#样品(3.48×10^7 CFU·mL^-1)(P<0.05)。不同预处理方式底泥微生物群落的AWCD值、六类碳源利用情况、多样性指数及主成分分析差异较大。240 h时,1#底泥方法4和方法5除对胺类碳源的利用(方法5显著高于方法4)外,微生物群落的上述其他指标差异均不显著(P>0.05),但有多项指标显著优于方法1、2、3(P<0.05);2#底泥方法5处理微生物群落的多项指标显著优于方法1~4;且无论在1#底泥还是2#底泥,方法5主成分分析的平行点均较为集中,说明其平行性较好。因此建议采用方法5进行养殖池塘底泥的预处理,即10%底泥稀释液在250 r·min^-1转速下振荡30 min,经600 r·min^-1离心10 min,离心上清液直接接入Biolog Eco板。

In order to explore the effects of sediment pretreatment methods on the results of Biolog EcoPlate analysis in aquaculture ponds, the sediments around and in the middle of the ponds of Penaeus vannamel were taken as research objects. The differences of average well color development (AWCD), utilization of six grouped carbon source, diversity indices and the principal component analysis of sediment microbial communities were examined under five pretreatment methods commonly used in literature. The results showed that the total number of cultivable bacteria in 1# sediment (7.73×10^5 CFU·mL^-1) were significant less than in 2# sediment(3.48×10^7 CFU·mL^-1)after 3 day of incubation(P<0.05). And there were large differences in AWCD values, utilization of six grouped carbon sources, diversity indices and principal component analysis of sediment microbial communities under different pretreatment methods. At 240 h of incubation, there was no significant differences in above indexes of 1# sediment between methods 4 and 5 except for the utilization of amine carbon sources (method 5 was significant higher than method 4), but many indexes in the two methods were significant superior to method 1, 2 and 3 (P<0.05);multiple indexes of 2# sediment treated by method 5 were significantly higher than those of method 1, 2, 3 and 4(P<0.05);the parallel points of method 5 in PCA plots were aggregated indicating that the data was stable no matter in 1# sediment or 2# sediment. The results suggested that method 5 was the optimal method for pretreating aquaculture pond sediment, and the method was as follows: 10 g of sediment were transferred to a conical flask containing 90 mL 0.85% sterilized saline, shaking for 30 minutes at 250 r·min^-1, the suspension was centrifuged for 10 minutes at 600 r·min^-1, and the centrifugal supernatant was directly inoculated into the Biolog EcoPlates.