Development of a real-time TaqMan assay to detect mendocina sublineage Pseudomonas species in contaminated metalworking fluids

Saha, Ratul; Donofrio, Robert; Bagley, Susan T.

doi:10.1007/s10295-010-0731-8

Cited by 13 publications

(12 citation statements)

References 25 publications

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“…These bacteria seem to be remarkably well adapted to aqueous MWFs, which can be explained by (i) the ubiquity of Pseudomonas spp. and their ability to grow in a large variety of media, especially water, (ii) a tolerance to the generally elevated pH of MWFs (34), and (iii) their ability to degrade hydrocarbons. Indeed, although a wide range of microorganisms have been found to utilize hydrocarbons as a sole source of carbon, most of the strains isolated in such media were Pseudomonas (43).…”

Section: Discussionmentioning

confidence: 99%

Factors Influencing the Microbial Composition of Metalworking Fluids and Potential Implications for Machine Operator's Lung

Murat

Grenouillet

Reboux

et al. 2012

Appl Environ Microbiol

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Hypersensitivity pneumonitis, also known as "machine operator's lung" (MOL), has been related to microorganisms growing in metalworking fluids (MWFs), especially Mycobacterium immunogenum. We aimed to (i) describe the microbiological contamination of MWFs and (ii) look for chemical, physical, and environmental parameters associated with variations in microbiological profiles. We microbiologically analyzed 180 MWF samples from nonautomotive plants (e.g., screw-machining or metalcutting plants) in the Franche-Comté region in eastern France and 165 samples from three French automotive plants in which cases of MOL had been proven. Our results revealed two types of microbial biomes: the first was from the nonautomotive industry, showed predominantly Gram-negative rods (GNR), and was associated with a low risk of MOL, and the second came from the automotive industry that was affected by cases of MOL and showed predominantly Gram-positive rods (GPR). Traces of M. immunogenum were sporadically detected in the first type, while it was highly prevalent in the automotive sector, with up to 38% of samples testing positive. The use of chromium, nickel, or iron was associated with growth of Gram-negative rods; conversely, growth of Gram-positive rods was associated with the absence of these metals. Synthetic MWFs were more frequently sterile than emulsions. Vegetable oil-based emulsions were associated with GNR, while mineral ones were associated with GPR. Our results suggest that metal types and the nature of MWF play a part in MWF contamination, and this work shall be followed by further in vitro simulation experiments on the kinetics of microbial populations, focusing on the phenomena of inhibition and synergy.

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Section: Discussionmentioning

confidence: 99%

Factors Influencing the Microbial Composition of Metalworking Fluids and Potential Implications for Machine Operator's Lung

Murat

Grenouillet

Reboux

et al. 2012

Appl Environ Microbiol

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“…Khan and Yadav (2004) developed and optimized SYBR Green qPCR assays for genus-specific detection and quantification of culturable and non-culturable mycobacteria and pseudomonads in MWFs. Saha et al (2010a) developed a TaqMan assay to detect mendocina sublineage Pseudomonas species in contaminated MWFs. The rapidly growing Mycobacterium species, mainly M. immunogenum, have been detected and quantified using TaqMan probe by Rhodes et al (2008Rhodes et al ( , 2011.…”

Section: Real-time Pcrmentioning

confidence: 99%

The microbiology of metalworking fluids

Saha

Donofrio

2012

Appl Microbiol Biotechnol

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Metalworking fluids (MWFs) are complex mixtures of chemicals and are indispensable materials in industry. They are used as cooling and lubricating agents in different machining process such as grinding, milling, and cutting. The quality of MWFs is affected by physical, chemical, and microbial contaminates. In particular, MWFs are highly vulnerable to microbial contamination, which may act both as potential pathogens and deteriorgens. Microbial contamination is of major concern due to potential health hazards such as skin dermatitis and hypersensitivity pneumonitis. The contaminated MWFs can exhibit high degrees of microbial loading, ranging from 10(4) to 10(10) colony-forming units (CFU)/ml. Wide varieties of microorganisms are reported to colonize MWFs. Traditional culturing techniques are not only laborious and time consuming but also underestimate the actual distribution of the microorganisms present in the contaminated MWFs. Therefore, rapid molecular methods such as real-time PCR and fluorescent in situ hybridization are implemented to monitor the microbial load. In industry, biocides are presently used to control microbial contamination. However, it has its own disadvantages and therefore, in recent years, alternative methods such as UV irradiation were evaluated to reduce microbial contamination in MWFs. Microbes inhabiting the MWF are also capable of forming biofilm which is detrimental to the MWF system. Biofilm is resistant to common disinfectant methods, and thus further research and development is required to effectively control its formation within MWF systems. This review is intended to discuss the overall microbiological aspects of MWF.

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“…lubricantis (ATCC BAA 1494) and P. fluorescens MWF-1 recovered in an earlier study (Saha et al 2010a) were selected for the study as they belong to the Pseudomonas rRNA group I. Escherichia coli (ATCC 23848) was selected as the negative control. All strains were grown and maintained on Tryptic Soy Agar (TSA) and Tryptic Soy Broth (TSB) (BD, Franklin Lakes, NJ).…”

Section: Bacterial Culturesmentioning

confidence: 99%

Rapid detection of rRNA group I pseudomonads in contaminated metalworking fluids and biofilm formation by fluorescent in situ hybridization

Saha

Donofrio

Goeres

et al. 2011

Appl Microbiol Biotechnol

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Metalworking fluids (MWFs), used in different machining operations, are highly prone to microbial degradation. Microbial communities present in MWFs lead to biofilm formation in the MWF systems, which act as a continuous source of contamination. Species of rRNA group I Pseudomonas dominate in contaminated MWFs. However, their actual distribution is typically underestimated when using standard culturing techniques as most fail to grow on the commonly used Pseudomonas Isolation Agar. To overcome this, fluorescent in situ hybridization (FISH) was used to study their abundance along with biofilm formation by two species recovered from MWFs, Pseudomonas fluorescens MWF-1 and the newly described Pseudomonas oleovorans subsp. lubricantis. Based on 16S rRNA sequences, a unique fluorescent molecular probe (Pseudo120) was designed targeting a conserved signature sequence common to all rRNA group I Pseudomonas. The specificity of the probe was evaluated using hybridization experiments with whole cells of different Pseudomonas species. The probe's sensitivity was determined to be 10(3) cells/ml. It successfully detected and enumerated the abundance and distribution of Pseudomonas indicating levels between 3.2 (± 1.1) × 10(6) and 5.0 (± 2.3) × 10(6) cells/ml in four different industrial MWF samples collected from three different locations. Biofilm formation was visualized under stagnant conditions using high and low concentrations of cells for both P. fluorescens MWF-1 and P. oleovorans subsp. lubricantis stained with methylene blue and Pseudo120. On the basis of these observations, this molecular probe can be successfully be used in the management of MWF systems to monitor the levels and biofilm formation of rRNA group I pseudomonads.

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Development of a real-time TaqMan assay to detect mendocina sublineage Pseudomonas species in contaminated metalworking fluids

Cited by 13 publications

References 25 publications

Factors Influencing the Microbial Composition of Metalworking Fluids and Potential Implications for Machine Operator's Lung

Factors Influencing the Microbial Composition of Metalworking Fluids and Potential Implications for Machine Operator's Lung

The microbiology of metalworking fluids

Rapid detection of rRNA group I pseudomonads in contaminated metalworking fluids and biofilm formation by fluorescent in situ hybridization

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