5种因子对赤红球菌HDRR2Y去除氨氮和亚硝酸盐效应的影响

徐煜; 胡晓娟; 张淞; 徐武杰; 苏浩昌; 文国樑; 曹煜成

doi:10.12131/20220044

5种因子对赤红球菌HDRR2Y去除氨氮和亚硝酸盐效应的影响

doi: 10.12131/20220044

徐煜^{1, 2,},
胡晓娟^{1, 2},
张淞¹,
徐武杰^{1, 2},
苏浩昌^{1, 2},
文国樑^{1, 2},
曹煜成^{1, 2, ,}

1.
中国水产科学研究院南海水产研究所/农业农村部南海渔业资源开发利用重点实验室/广东省渔业生态环境重点实验室，广东广州 510300
2.
中国水产科学研究院南海水产研究所深圳试验基地，广东深圳 518121

基金项目: 国家重点研发计划项目 (2020YFD0900401)；广东省重点领域研发计划项目 (2021B0202040001)；海南省自然科学基金项目 (2022CXTD265)；中国水产科学研究院基本科研业务费项目 (2020TD54)；中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金 (2021SD08)；国家现代农业产业技术体系 (CARS-48)；广东省现代农业产业技术体系创新团队建设专项资金 (2019KJ149)

详细信息

作者简介:
徐煜：徐　煜 (1987—)，男，助理研究员，硕士，研究方向为对虾养殖技术与水质调控。E-mail: xuyublq@163.com

通讯作者:
曹煜成 (1979—)，男，研究员，博士，研究方向为水产健康养殖及养殖生态环境调控与修复。E-mail: cyc_715@163.com

中图分类号: S 917.1
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- 文章访问数: 310
- HTML全文浏览量: 53
- PDF下载量: 14
- 被引次数: 0
出版历程
- 收稿日期: 2022-02-24
- 修回日期: 2022-06-13
- 录用日期: 2022-07-19
- 网络出版日期: 2022-07-27
- 刊出日期: 2023-02-03

Effect of five factors on removing ammonia nitrogen and nitrite by Rhodococcus ruber HDRR2Y fermentation

XU Yu^{1, 2
,},
HU Xiaojuan^{1, 2},
ZHANG Song¹,
XU Wujie^{1, 2},
SU Haochang^{1, 2},
WEN Guoliang^{1, 2},
CAO Yucheng^{1, 2
, ,}

1.
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences/Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs/Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, Guangzhou, 510300, China
2.
Shenzhen Base of South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shenzhen, 518121, China

摘要

摘要: 赤红球菌 (Rhodococcus ruber) 是常见的污水、废水处理微生物，为探究环境因子对赤红球菌HDRR2Y去除氨氮 (${\rm{NH}}_4^{^{ +}} $-N)和亚硝酸盐 (${\rm{NO}}_2^{^{ -}} $-N) 效应的影响，将HDRR2Y菌株发酵液添加至含有高浓度${\rm{NH}}_4^{^{ +}} $-N或${\rm{NO}}_2^{^{ -}} $-N的养殖水体中，通过国标法检测氨氮和亚硝酸盐浓度的变化，检验菌株培养液去除氨氮及亚硝酸盐的效果。按照温度、转速、盐度、接种菌量、底物浓度 (氨氮和亚硝酸盐) 5个因素进行Plackett-Burman实验设计，探讨这些因子对赤红球菌去除氨氮和亚硝酸盐效应的影响。结果显示，发酵过程中，菌株HDRR2Y浓度在36 h内从初始的5×10⁴ CFU·mL⁻¹增至4.08×10⁹ CFU·mL⁻¹。添加菌株培养液后，养殖水体氨氮质量浓度从初始的15 mg·L⁻¹降至5.56 mg·L⁻¹，去除率为62.96%；亚硝酸盐质量浓度从15 mg·L⁻¹降至6.95 mg·L⁻¹，去除率为59.37%。5种因子中，温度和氨氮浓度对菌株HDRR2Y去除氨氮影响最显著 (P<0.05)，影响权重程度依次为：温度>氨氮浓度>转速>菌量>盐度；温度和转速是影响去除亚硝酸盐最显著的两个因子 (P<0.05)，权重程度依次为：温度>转速>盐度>亚硝酸盐浓度>菌量。可见，菌株HDRR2Y培养液具有良好的去除氨氮、亚硝酸盐的作用，且温度是影响HDRR2Y去除氨氮、亚硝酸盐效率的最显著因子。
- 赤红球菌 /
- 氨氮 /
- 亚硝酸盐 /
- 去除率 /
- 环境因子
Abstract: The bacteria Rhodococcus ruber is usually used in waste water and sewage treatment. In order to discuss the effect of environmental factors on R. ruber HDRR2Y's removing ammonia nitrogen and nitrite, we added the culture medium of R. ruber strain HDRR2Y to the aquaculture water containing high concentration of ammonia nitrogen (NH₄ ⁺-N) and nitrite (NO₂ ⁻-N), and observed the changes of ammonia nitrogen and nitrite concentrations by the national standard method, so as to investigate the effect of the culture medium on the removal of ammonia nitrogen and nitrite. In addition, we carried out the Plackett-Burman experimental design according to five factors: temperature, rotating speed, salinity, inoculum amount, substrate (Ammonia nitrogen and nitrite) concentration, to explore the effect of these factors on the removal of ammonia nitrogen and nitrite by R. ruber. The results show that during the fermentation process, the mass concentration of strain HDRR2Y increased from 5×10⁴ CFU·mL⁻¹ to 4.08×10⁹ CFU·mL⁻¹ in the initial 36 h. After adding the culture medium, the mass concentration of ammonia nitrogen decreased from 15 mg·L⁻¹ to 5.56 mg·L⁻¹, with a removal rate of 62.96%, and that of nitrite decreased from 15 mg·L⁻¹ to 6.95 mg·L⁻¹, with a removal rate of 59.37%. Among the five factors, temperature and ammonia nitrogen concentration had the most significant effects on the removal of ammonia nitrogen by strain HDRR2Y (P<0.05) (Temperature>ammonia nitrogen concentration>rotation speed>biomass>salinity). Temperature and rotating speed were the two most significant factors affecting the removal of nitrite (P<0.05) (Temperature>rotating speed>salinity>nitrite concentration>bacterial count). It is showed that the culture medium of strain HDRR2Y is good for removing ammonia nitrogen and nitrite, and temperature is the most significant factor affecting the efficiency of HDRR2Y in removing ammonia nitrogen and nitrite.
- Rhodococcus ruber /
- Ammonia nitrogen /
- Nitrite /
- Removal rate /
- Environment factors

HTML全文

图 1 菌株HDRR2Y菌浓度变化

Figure 1. Bacterial concentration change of strain HDRR2Y

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图 2 菌株HDRR2Y在发酵过程中的形态

Figure 2. Morphology of strain HDRR2Y during fermentation

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图 3 赤红球菌HDRR2Y对养殖水体氨氮和亚硝酸盐作用效果

Figure 3. Effect of HDRR2Y on ammonia and nitrite nitrogen in aquaculture water

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表 1 Plackett-Burman实验因素及与水平设计

Table 1. Factors and levels design of Plackett-Burman

因素 Factor	水平 Level
因素 Factor	−1	1
A：菌浓度 Bacterial concentration/(CFU·mL⁻¹)	10⁴	10⁷
B：底物 (氨氮/亚硝酸盐) 质量浓度 Substrate concentration (Ammonia nitrogen/Nitrite)/(mg·L⁻¹)	15	30
C：盐度 Salinity/‰	15	30
D：转速 Rotating speed/(r·min⁻¹)	150	225
E：温度 Temperature/℃	20	30

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表 2 Plackett-Burman设计

Table 2. Plackett-Burman experimental design

序号 No.	A：菌浓度 Bacterial concentration	B：底物 (氨氮/ 亚硝酸盐) 浓度 Substrate concentration (Ammonia nitrogen/Nitrite)	C：盐度 Salinity	D：转速 Rotating speed	E：温度Temperature
1	1	−1	−1	−1	1
2	1	−1	1	1	1
3	1	−1	1	1	−1
4	−1	1	−1	1	1
5	−1	1	1	−1	1
6	−1	−1	−1	1	−1
7	−1	−1	1	−1	1
8	1	1	−1	−1	−1
9	−1	−1	−1	−1	−1
10	−1	1	1	1	−1
11	1	1	−1	1	1
12	1	1	1	−1	−1

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表 3 养殖水体中氨氮和亚硝酸盐去除率

Table 3. Removal rate of ammonia nitrogen and nitrite in aquaculture water

被测指标 Measured index	时间 Time	去除率 Removal rate/%
被测指标 Measured index	时间 Time	对照组 GC	加菌组 GH
氨氮 NH₄ ⁺-N	第3天	−5.16±6.87^a	62.92±1.53^b
氨氮 NH₄ ⁺-N	第7天	−4.06±7.31^a	57.50±2.16^b
亚硝酸盐 NO₂ ⁻-N	第3天	−1.43±2.10^a	59.37±2.49^b
亚硝酸盐 NO₂ ⁻-N	第7天	−4.92±1.41^a	56.36±1.67^b
注：同行数据的不同字母代表对照组和加菌组差异显著 (P<0.05)。 Note: Values with different letters within the same column indicate significant difference (P<0.05).

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表 4 Plackett-Burman实验底物去除率

Table 4. Removal rate of substrate in Plackett-Burman experiment

序号 No.	Y₁：氨氮去除率 Removal rate of ammonia nitrogen/%	Y₂：亚硝酸盐去除率 Removal rate of nitrite/%
1	48.4	42.5
2	51.3	36.6
3	36.2	40.2
4	52.4	47.1
5	53.6	36.8
6	42.5	42.9
7	43.9	39.9
8	40.7	32.5
9	31.6	43.7
10	43.8	38.7
11	59.3	53.4
12	45.2	30.7

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表 5 Plackett-Burman实验因素水平与回归方程方差分析 (底物为氨氮)

Table 5. Plackett-Burman's experimental factor level and ANOVA of regression equation (Substrate is ammonia nitrogen)

因素 Factor	平方和 Sum of squares	自由度 df	均方 Mean square	F	P>F	显著性 Significance	重要性排序 Importance ranking
模型 Model	591.88	5	118.38	10.09	0.007 0	**
A：菌浓度 Bacterial concentration/(CFU·mL⁻¹)	14.74	1	14.74	1.26	0.305 2		4
B：氨氮质量浓度 Ammonia nitrogen mass concentration/ (mg·L⁻¹)	140.77	1	140.77	12.00	0.013 4	*	2
C：盐度 Salinity/‰	0.0675	1	0.067 5	0.005 8	0.942 0		5
D：转速 Rotating speed/(r·min⁻¹)	40.70	1	40.70	3.47	0.111 9		3
E：温度 Temperature/℃	395.60	1	395.60	33.71	0.001 1	*	1
残留误差 Residual error	70.41	6	11.74
总差 Total difference	662.29	11
注：判定系数R²=0.893 7；. 差异显著 (P<0.05)；. 差异极显著( P<0.001)。 Note: Determination coefficient (R²)= 0.893 7; . Significant difference (P<0.05); **. Very significant difference (P<0.001).

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表 6 Plackett-Burman实验因素水平与回归方程方差分析 (底物为亚硝酸盐)

Table 6. Plackett-Burman's experimental factor level and ANOVA of regression equation (Substrate is nitrite)

因素 Factor	平方和 Sum of squares	自由度 df	均方 Mean square	F	P>F	显著性 Significance	重要性排序 Importance ranking
模型 Model	340.26	5	68.05	4.96	0.038 2	*
A：菌浓度 Bacterial concentration/(CFU·mL⁻¹)	7.05	1	7.05	0.514 5	0.500 2		5
B：氨氮质量浓度 Ammonia nitrogen mass concentration/(mg·L⁻¹)	8.00	1	8.00	0.583 8	0.473 8		4
C：盐度 Salinity/‰	11.21	1	11.21	0.817 9	0.400 6		3
D：转速 Rotating speed/(r·min⁻¹)	152.65	1	152.65	11.13	0.015 7	*	2
E：温度 Temperature/℃	161.33	1	161.33	11.77	0.014 0	*	1
残留误差 Residual error	82.26	6	13.71
总差 Total difference	422.52	11
注：判定系数R²=0.8053；. 差异显著 (P<0.05)。 Note: Determination coefficient (R²)= 0.8053; . Significant difference (P<0.05).

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