Table 1-1: List of chemical substance families and their main function in metalworking fluids, adapted from Müller (2000) ... 15 Table 1-2: Cost calculation for MWF related expenses (adapted from Betrieblicher Umweltschutz in BW, 2010) ... 18 Table 2-1: Protocol for 16SrDNA PCR of whole bacterial DNA ... 31 Table 2-2: Species used for inoculating MWF and in vitro biofilm development in MWF with characteristics: Gram staining (positive [+], negative [-], variable [+/-]), motility (motile [+], immobile [-]), spore formation (able [+], unable [-]) ... 35 Table 2-3: Influence of different incubation conditions on the in vitro biofilm development, the matrix shows the different variation using the example of stainless steel;
the same design of experiments was carried out with the aluminum alloy, copper and the glass slides. 37
Table 2-4: Selection of species along with a short phenotype description, used for MWF degradation experiments, the bold printed species were used for single culture testing, two different co-cultures were mixed: mix of five with the bold printed species and a mix of ten with all listed species ... 39 Table 2-5: MWF compounds with effects and concentration used in degradation experiments with bacteria isolated from industrially used MWF, stock concentration states the percentage of compound in MWF concentrates. The final concentration used for experiments is stated in brackets. ... 39 Table 3-1: Properties of biofilms from five different machines; the characterization includes the biofilm density and the water content, the biodiversity of the biofilm associated bacteria along with the corresponding risk group classification stating the risk group and the number of isolated bacteria in brackets. The microbial burden of the MWF in question is given and the biodiversity of it with the risk group classification of the isolates. ... 58 Table 3-2: Evaluation of the in vitro biofilm system by comparison with the real system and a known biofilm builder (Pseudomonas pseudoalcaligenes), the biofilm formation was carried out on glass surfaces, so as to have an inert surface. ... 61 Table 3-3: Average microbial burden in the MWF used in the in vitro system after seven days, in relation to the different incubation conditions and the metal substratum ... 67 Table 3-4: Arithmetic surface roughness Sa of the metal substrata used for biofilm formation before and after biofilm formation, measured area: 10µm x 10µm. The surface was cleaned prior to measurement. ... 70 Table 3-5: Comparison of degradation of MWF compounds by planktonic and biofilm associated bacteria. (-) indicates no detectable growth; (+) abundance smaller than |0,5|;
(++) abundance between |0,5| and |1,5|; (+++) abundance higher than |1,5|. The most efficiently degrading culture is stated in brackets. ... 76 Table 10-1: Published bioburden in CFU/ml or cells/ml detected in different MWFs ... 120 Table 10-2: Summary of microorganisms, isolated from liquid MWF, published in peer reviewed journals ... 121
Table 10-3: Summary of all identified isolates from the five MWFs in examination.
Species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in %. If not stated, the identification was based on macroscopic and microscopic characterization and staining
behavior. 126
Table 10-4: Isolated and identified airborne microorganisms, isolated from the area around machine SS-M1 and SS-M2, species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in %. If not stated, the identification was based on macroscopic and microscopic characterization and staining behavior. ... 127 Table 10-5: Isolated and identified airborne microorganisms, isolated from the area around machines PS-M1, PS-M2 and PS-M3, the species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in %. If not stated, the identification was based on macroscopic and microscopic characterization and staining behavior. ... 128 Table 10-6: Identified isolates from the preparation water of MWF used in small work hall. The species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in % ... 129 Table 10-7: Microorganisms isolated from contact plates taken from unprocessed and processed metals, the species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in %. If not stated the identification was based on macroscopic and microscopic characterization and staining behavior. “Alu” is short for aluminum alloy; ... 130 Table 10-8: Description and identification of bacterial colonies after direct culturing on TSA for ten days at RT, the species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in %. If not stated, the identification was based on macroscopic and microscopic characterization and staining behavior. ... 131 Table 10-9: Summary of all biofilm isolates from the 1st set of coupons depending on the sort of plastic used ... 132 Table 10-10: Identified isolates from cleaning the MWF tank of machine SS-M1 during MWF exchange, the species identification is based on the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in %. If not stated the identification was based on macroscopic and microscopic characterization and staining behavior. ... 132
Table 10-11: Results from the second set of biofilm experiment with short sampling intervals. Both the biofilm and the MWF were sampled on the listed sampling dates. Species identification is based on the macroscopic and the microscopic phenotype, Gram characteristics and the sequence of the 16S rDNA PCR Fragment. The length of the fragment is given in base pairs and the similarity to 16S rDNA genes found in NCBI collections in Ident %. If not stated the identification was based on macroscopic and microscopic characterization and staining behavior. ... 133 Table 10-12: List of isolated biofilm associated microorganisms from the MWF systems of industrial site 1. Species are characterized by microscopic and macroscopic properties and by their 16S rDNA sequence. Ident states the percentage of similarity with a given species in a 16S rDNA library, bp states the fragment length used for alignment. RG states the risk group for microorgansims classification in accordance to the WHO and the German
“technische Regeln für biologische Arbeitstoffe TRBA 466” ... 135 Table 10-13: List of isolated biofilm associated microorganisms from the MWF systems of industrial site 2. Species are characterized by microscopic and macroscopic properties and by their 16S rDNA sequence. Ident states the percentage of similarity with a given species in a 16S rDNA library, bp states the fragment length used for alignment. RG states the risk group for microorgansims classification in accordance to the WHO and the German
“technische Regeln für biologische Arbeitstoffe TRBA 466” ... 137 Table 10-14: List of isolated and identified microorganisms from MWFs of five different machines, Part A industrial site 1. Species are characterized by microscopic and macroscopic properties and by their 16S rDNA sequence. Ident states the percentage of similarity with a given species in a 16S rDNA library, bp states the fragment length used for alignment. RG states the risk group for microorgansims classification in accordance to the WHO and the German “technische Regeln für biologische Arbeitstoffe TRBA 466” ... 138 Table 10-15: Isolated and identified microorgansims from MWF of five different machines, Part B industrial site 2. Species are characterized by microscopic and macroscopic properties and by their 16S rDNA sequence. Ident states the percentage of similarity with a given species in a 16S rDNA library, bp states the fragment length used for alignment. RG states the risk group for microorgansims classification in accordance to the WHO and the German “technische Regeln für biologische Arbeitstoffe TRBA 466” ... 139 Table 10-16: Results of the biofilm characterization of the biofilm development in incubation variations after seven days incubation ... 140