Diversity Research of Microbial in Water-based Metalworking Fluids
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摘要: 微生物污染会缩减金属加工液使用寿命，细菌比真菌更容易在金属加工液中滋生。在所采集的6组金属加工液样本中，共检测出细菌2门、2纲、5目、6科、10属和14种，而真菌为4门、8纲、10目、14科、15属和17种，真菌多样性更丰富，但真菌只出现在细菌污染严重的样本中。种水平下，弗氏柠檬酸杆菌(Citrobacter_freundii)、Unclassified_g_Citrobacter、Unclassified_f_Enterobacteriaceae被确定为优势细菌，且检出细菌以革兰氏阴性菌为主；真菌以未知真菌(Unclassified_k_Fungi)和Fusarium_petroliphilum为主。Abstract: The service life of metalworking fluids can be shortened by existence of microorganism, and it is necessary to explore the composition of microbial communities in metalworking fluids. Three different methods were measured to determine the best one to separate microorganisms from metal working fluids. The concentration of microorganisms can be increased by the mikrocount combi method which has the optimal separation result. Under the help of Illumina MiSeq high-throughput sequencing, the composition of the microbial diversity of metalworking fluid samples at the 6 levels of phylum, class, order, family, genus, and species were completed respectively. Moreover, bacteria were detected in six groups of samples, while fungus were discovered in only two groups. Meanwhile, it was easier for bacteria to thrive in metalworking fluids than fungi, and fungi was only existed in samples with high bacterial contamination. A total of 2 phyla, 2 classes, 5 orders, 6 families, 10 genera and 14 species of bacteria were detected in all samples, while 4 phyla, 8 classes, 10 orders, 14 families, 15 genera and 17 species of fungi were also detected, which means that the fungal diversity is more abundant. Citrobacter_freundii_g_Citrobacter, unclassified_g_Citrobacter, unclassified_f_Enterobacteriaceae were identified as the dominant bacteria, and most of bacteria detected were Gram-negative. The composition of the metalworking fluid will affect the type of bacteria. All detected bacteria can destroy the stability of the metal working fluid through different ways, which shorten its service life. The dominant fungi were unclassified_k_Fungi and Fusarium_petroliphilum. The health of operators will be harmed by metalworking fluids with microbial contamination.
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Key words:
- metalworking fluids /
- microorganism /
- diversity /
- bacteria /
- fungi
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图 1 不同水平下金属加工液细菌群落组成
Figure 1. Bacterial composition in metalworking fluid at different level
图 2 属水平下金属加工液细菌群落组成
Figure 2. Bacterial composition in metalworking fluid at genus level
图 3 种水平下金属加工液细菌群落组成
Figure 3. Bacterial composition in metalworking fluid at species level
图 4 不同水平下金属加工液真菌群落组成
Figure 4. Fungal composition in metalworking fluid at different level
图 6 种水平下金属加工液真菌群落组成
Figure 6. Fungal composition in metalworking fluid at species level
表 1 样本采集信息
Table 1. Details of sample collection
Sample Collection date Collection location Temperature/ ℃ Usage time/ d pH COD/ (mg·L−1) QX1 2020/01 Minhang, Shanghai 3~9 90 7.88 60429 QX2 2020/06 Hefei, Anhui 23~31 300 6.34 81417 QX3 2020/05 Weifang, Shandong 12~26 150 6.59 78816 CJ 2021/03 Wuhu, Anhui 8~17 180 6.95 66242 JX 2021/03 Lishui, Zhejiang 10~20 450 5.72 80538 WF 2020/08 Weifang, Shandong 23~32 150 7.42 113574 
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表 2 分离前后不同样品细菌浓度
Table 2. Bacterial concentration of different samples before and after separation
Sample concentration / (CFU·mL−1) Original emulsion 104 Sample after PAC demulsification 10 Sample after centrifugation 104 Sample after mikrocount combi method 106
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表 3 金属加工液微生物多样性统计
Table 3. Diversity of microbial in water-based metal working fluids
Sample Microorganism Phylum Class Order Family Genus Species OTU QX1 Bacteria 1 1 1 2 2 2 3 Fungi / / / / / / / QX2 Bacteria 1 1 2 3 5 5 5 Fungi 4 7 9 12 13 14 23 QX3 Bacteria 1 1 2 3 4 4 5 Fungi / / / / / / / CJ Bacteria 1 1 2 2 3 3 3 Fungi / / / / / / / JX Bacteria 2 2 4 4 4 4 5 Fungi 4 6 6 6 7 7 8 WF Bacteria 2 2 4 5 5 7 9 Fungi / / / / / / /
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