UPM常熟纸厂PM2网下白水中微生物的分离和鉴定
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摘要
当微生物生长时,常常趋向于附着于物体表面从而形成生物膜,也就是造纸上说的沉积物。沉积物的形成是造成纸机生物膜污染问题的重要原因,这一问题也越来越引起人们的重视。然而在纸机的白水系统中,引起生物膜污染的微生物种类和特性并未得到很好的研究。
为了确定引起纸机生物膜污染的主要微生物,从而揭示引起生物膜污染的根本原因。以UPM常熟纸厂PM2网下白水为试样,采取常规纯培养的方法,获得优势菌株20株,根据细菌的菌落形态、菌落颜色、革兰氏染色等常见特征,从中选取4株差别比较明显的菌株(编号分别为N1、N2、N4、N16)进行进一步研究。通过16S rDNA序列进行分类研究,确定该细菌的分类地位,并结合个体形态与培养特征、常规生理生化特性进行鉴定,确定各细菌所属种。实验表明:N1、N2为肺炎克雷伯氏菌(Klebsiella pneumoniae),其中N2为肺炎克雷伯氏菌鼻硬结亚种(Klebsiella pneumoniae Rhinoscleroma);N4为鞘氨醇单胞菌属中的一种Sphingomonas azotifigens;N16为肠杆菌属中的一种Enterobacter radicincitans。鉴于菌株N1、N2相对N4、N16有较厚的荚膜,由此推断N1、N2,即肺炎克雷伯氏菌种,可能是造成高速纸机运转过程中产生生物膜污染的主要菌种。这一结果对研究造纸过程中生物膜污染问题提供了积极的指导意义。
肺炎克雷伯氏菌作为一种条件致病菌,其分布较广,且由于产酶和生物被膜等特性以及抗生素的大量使用,导致其抗药性较强。如何抑制其生长,亟待更深一步的研究。以上结果对研究和解决造纸过程中生物膜污染问题提供了积极的指导意义。
When microbial growth, they often tend to attach to the surface to form a biofilm, which is called the sediment in paper-making industry. The formation of sediment is the main reason that results in microbial contamination of the paper machine, and this problem has drawn increasing attention by more and more people. However, which kind of microbial specie(s) and how they work in the white water system has not been well known.
In order to find the main microorganism which produce the microbial biofilm and then find the root causes of microbial contamination. White water of the wire pit was sampled from the PM2 of UPM(Changshu) Mill. By the conventional pure culture method, 20 dominant microbial strains were selected. Thereafter, 4 strains of these microorganisms, which were obviously different in shape, color and some other common characteristics, were used to conduct further analyses (Serial No.: N1、N2、N4、N16). The phylogenetic relationship of the 4 strains was determined by sequencing the 16S rDNA. Taxonomy of these strains was also studied through physiological and chemical tests, and morphorlogical observation. The experiments indicated: N1 and N2 were Klebsiella pneumonia, N2 was Klebsiella pneumoniae Rhinosclerom, additionally; N4 was Sphingomonas azotifigens; N16 was Enterobacter radicincitans. The thicker capsule of strain N1 and N2 comparing with strain N4 and N16 implied that N1, N2 were Klebsiella pneumonia, which likely to be the main disease bacterium that brought about pollution to biofilm of the paper machine. This study provided useful guidance for analyzing the causes for biofilm-contamination in papermaking process.
As a kind of opportunistic pathogen, Klebsiella pneumonia has a widely distribution and a strong resistance, which because of its producing-enzyme and biofilms, as well as the extensive use of antibiotics These results provides a positive guide for analyze the reason of biofilm-contamination in papermaking process.
为了确定引起纸机生物膜污染的主要微生物,从而揭示引起生物膜污染的根本原因。以UPM常熟纸厂PM2网下白水为试样,采取常规纯培养的方法,获得优势菌株20株,根据细菌的菌落形态、菌落颜色、革兰氏染色等常见特征,从中选取4株差别比较明显的菌株(编号分别为N1、N2、N4、N16)进行进一步研究。通过16S rDNA序列进行分类研究,确定该细菌的分类地位,并结合个体形态与培养特征、常规生理生化特性进行鉴定,确定各细菌所属种。实验表明:N1、N2为肺炎克雷伯氏菌(Klebsiella pneumoniae),其中N2为肺炎克雷伯氏菌鼻硬结亚种(Klebsiella pneumoniae Rhinoscleroma);N4为鞘氨醇单胞菌属中的一种Sphingomonas azotifigens;N16为肠杆菌属中的一种Enterobacter radicincitans。鉴于菌株N1、N2相对N4、N16有较厚的荚膜,由此推断N1、N2,即肺炎克雷伯氏菌种,可能是造成高速纸机运转过程中产生生物膜污染的主要菌种。这一结果对研究造纸过程中生物膜污染问题提供了积极的指导意义。
肺炎克雷伯氏菌作为一种条件致病菌,其分布较广,且由于产酶和生物被膜等特性以及抗生素的大量使用,导致其抗药性较强。如何抑制其生长,亟待更深一步的研究。以上结果对研究和解决造纸过程中生物膜污染问题提供了积极的指导意义。
When microbial growth, they often tend to attach to the surface to form a biofilm, which is called the sediment in paper-making industry. The formation of sediment is the main reason that results in microbial contamination of the paper machine, and this problem has drawn increasing attention by more and more people. However, which kind of microbial specie(s) and how they work in the white water system has not been well known.
In order to find the main microorganism which produce the microbial biofilm and then find the root causes of microbial contamination. White water of the wire pit was sampled from the PM2 of UPM(Changshu) Mill. By the conventional pure culture method, 20 dominant microbial strains were selected. Thereafter, 4 strains of these microorganisms, which were obviously different in shape, color and some other common characteristics, were used to conduct further analyses (Serial No.: N1、N2、N4、N16). The phylogenetic relationship of the 4 strains was determined by sequencing the 16S rDNA. Taxonomy of these strains was also studied through physiological and chemical tests, and morphorlogical observation. The experiments indicated: N1 and N2 were Klebsiella pneumonia, N2 was Klebsiella pneumoniae Rhinosclerom, additionally; N4 was Sphingomonas azotifigens; N16 was Enterobacter radicincitans. The thicker capsule of strain N1 and N2 comparing with strain N4 and N16 implied that N1, N2 were Klebsiella pneumonia, which likely to be the main disease bacterium that brought about pollution to biofilm of the paper machine. This study provided useful guidance for analyzing the causes for biofilm-contamination in papermaking process.
As a kind of opportunistic pathogen, Klebsiella pneumonia has a widely distribution and a strong resistance, which because of its producing-enzyme and biofilms, as well as the extensive use of antibiotics These results provides a positive guide for analyze the reason of biofilm-contamination in papermaking process.
引文
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