art 7 81-92
art 7 81-92
Grzegorz Kurowski
Otmar Vogt ([email protected])
Jan Ogonowski
Faculty of Chemical Engineering and Technology, Cracow University of Technology
Paint-degrading microorganisms
Abstract
The paper presents a literature survey of research on the biodeterioration and biodegradation of paint
coatings, as well as study findings on the biodegradation of varnish coatings and agents used for their removal.
Keywords: biodeterioration, biodegradation, paints, microorganisms
Streszczenie
W artykule przedstawiono przegląd literaturowy dotyczący biodeterioracji i biodegradacji powłok
malarskich. Przedstawiono również badania dotyczące biodegradacji substancji używanych do usuwania
powłok lakierniczych.
Słowa kluczowe: kluczowe: biodeterioracja, biodegradacja, powłoka malarska, mikroorganizmy
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1. Introduction
Paint and varnish coatings are used for protective and/or decorative purposes, and their
properties largely depend on their ingredients, i.e. on the film-forming substances employed.
The latter primarily include macromolecular organic substances or substances that turn into
macromolecular compounds by way of reactions that accompany the formation of a varnish
layer. These substances also determine the possible uses and applications of paint coatings.
Film-forming substances consist of resins and other accessory substances. In order to invest
coatings with specific properties a mix of resins is also often used [1].
The literature distinguishes between the processes of biodeterioration and biodegradation.
Biodeterioration involves the destruction of an economically important substance by
microorganisms. The term is often used to refer to a deterioration in the properties of substances
that are normally immune to microbiological attack, such as metals, plastics, medicines,
cosmetics, paints and varnishes, fuels, oils, and other objects. Biodeterioration is thus an
adverse phenomenon. In contrast, biodegradation is viewed as beneficial. The biodeterioration
of varnish coatings caused by microorganisms is often accompanied by the deterioration of
their protective and decorative properties. The proliferation of microorganisms on and inside
paint may cause it to peel. The growth of fungi, in turn, may lead to discolorations [2, 3].
The microbiological resistance of resins used in varnishes can be measured on the
Wasserbauer scale, which assesses the proliferation of microorganisms on a scale from 0 to 3,
where 3 indicates robust growth. Varnishes with the index of 2 or more are described as non-
resistant to microorganisms. The values of microbiological resistance for selected synthetic
resins are as follows: acrylic resin 0.51; phenol formaldehyde resin 0.73; urea-formaldehyde
resin 0.17; polyester resin 1.20; epoxy resin 0.19; polyvinyl resin 0.51 [4].
2.1. Biodeterioration
Only water-based paints are susceptible to biodeterioration. The cellulose ethers they
contain as thickening agents are often attacked by enzymes produced by fungi and bacteria,
which enter the paint at the manufacturing stage together with contaminated ingredients [3].
Orehek et al. studied the biodeterioration of carboxymethyl cellulose by Bacillus subtilis subsp.
subtilis NCIB 3610. The bacteria turned the original pseudo-plastic non-Newtonian fluid that
contained carboxymethyl cellulose into a Newtonian fluid. In addition, the fluid’s viscosity
dropped from 10 to 1.4 mPas and the concentration of reducing sugars increased eighteen-
fold. Bacillus subtilis subsp. subtilis NCIB 3610 was also shown to synthesize cellulases that
effectively break down chemical bonds in carboxymethyl cellulose; however, it is not able to
reutilize the end products of hydrolysis [5].
Obidi et al. [2009] studied the microbiological contamination of painting products. Fresh
samples were taken to isolate the following bacterial strains: Bacillus brevis, Bacillus laterosporus,
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Bacillus polymyxa, Lactobacillus gasseri, Lactobacillus brevis, Proteus mirabilis, Escherichia coli and
fungi Aspergillus niger, Aspergillus flavus, and Penicillium citrinum. Monitored over a period of 10
months, the population of isolated microorganisms increased. For bacteria, the population grew
from 1.6∙101 cfu∙cm-3 in a fresh sample to 4.7∙105 cfu∙cm-3 10 months later. The corresponding
figures for fungi were 1.0∙101 and 5.5∙103 cfu∙ cm-3, respectively [6]. In addition, the researchers
took samples of water-based paints to isolate a strain of Pseudomonas aeruginosa [7].
Shirakawa et al. (2011) studied microorganisms living in four different acrylic paints
exposed to atmospheric conditions for seven years in Sao Paulo and the coastal town of
Ubatuba in Brazil. They identified bacteria from the cyanobacteria group. The main strains
present on the surface of all four types of paint belonged to Gloeocapsa and Scytonema.
In addition, the following genera were detected: Synechocystis, Synechococcus, Xenococcus,
Chroococcidiopsis, Myxosarcina, Oscillatoria, Leptolyngbya, Chlorella, Stichococcus, Coccoid
chlorophyte, Klebsormidium. The researchers also observed certain differences depending on
the location from which the samples were obtained. Paint degradation related to the presence
of biofilm was shown to be very low, but progressed faster in Ubatuba [8]. Other researchers
also isolated cyanobacteria from painted buildings in Latin America [9-11]. Stenotrophomonas
maltophilia were also detected in water-based acrylic paints [12].
De Souza and Gaylarde (2002) isolated microorganisms from varnished wood panels
exposed to atmospheric conditions for two years in South Brazil. The following bacteria
were isolated from the samples: Corynebacterium sp., Bacillus sp., as well as bacteria from the
Enterobacteriaceae family. Yeasts were also detected: Rhodotorula lactosa, Exophiala jeanselmei,
as well as fungi: Aureobasidium sp., Penicillium sp., Paecilomyces sp. [13].
Pangallo et al. (2015) investigated the biodeterioration of epoxy resins. The most frequently
isolated bacteria present on surfaces coated with epoxy resins belonged to the Firmicutes
phylum, especially the genus of Bacillus. γ-Proteobacteria, and particularly Pseudomonas, were
also frequently detected [14]. Similar results were obtained by Cappitelli et al. [15], who
studied surfaces coated with epoxy resins and isolated Gram-positive bacteria of the Bacillus,
Brevibacillus, Micrococcus, Staphylococcus and Kocuria genus, as well as Gram-negative bacteria
from the genus of Pseudomonas, Agrobacterium and Ochrobacter [15]. In addition, researchers
[14] identified several species of black yeast: Friedmanniomyces endolithicus, Pseudotaeniolina
globosa, Phaeococcomyces catenatus and Catenulostroma germanicum.
The authors of paper [16] listed the following species of microorganisms: Aspergillus
versicolor, Aspergillus niger, Cladosporium herbarum, Cladosporium sphaerospermum,
Engyodontium album, Penicillium aurantiogriseum, Penicillium chrysogenum, Trichoderma
longibrachiatum, Debaryomyces hansenii, Rhodotorula mucilaginosa, Bacillus subtilis, Bacillus
licheniformis and Rhodococcus fascians, all isolated from paints [16].
Bacteria present in paint on stone walls were studied by Gurtner et al. [17]. The researchers
analysed mural paintings in the chapel of an Austrian castle and a German church. The following
bacteria were isolated: bacteria from the phylum of Actinobacteria, such as Arthrobacter
sp., Actinobispora sp., Amycolata sp., Asiosporangium sp., Frankia sp., Geodermatophilus sp.,
Nocardioides sp., Promicromonospora sp., Pseudonocardia sp., Rubrobacter sp., Streptomonospora sp.,
Saccharopolyspora sp., Sphaerobacter sp. and Thermocrismum sp., bacteria from the Proteobacteria
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phylum, such as Aquaspirillum sp., Chromohalobacter sp., Deleya sp., Erythrobacter sp., Halomonas
sp., Porphyrobacter sp., Pseudomonas sp., Rhizobium sp., Salmonella sp. In addition, the researchers
isolated bacteria from the genera of Bacillus, Paenibacillus, Micrococcus, Staphylococcus,
Methylobacterium, and Halomonas [17]. Similar results were obtained by the authors of articles
[18, 19], who studied mural paintings in Spain and Italy. Apart from the phyla mentioned above,
researchers who wrote papers [20, 21] also isolated Cyanobacteria (Chlorogloea microcystoides,
Chroococcus lithophilus, Gloeocapsa spp., Gloeothece rupestris, Pseudocapsa dubia). Bacteria
from the Actinobacteria phylum of the Pseudonocardia, Streptomyces, Nocardia, Rhodococcus,
Nocardioides, Amycolatopsis, Saccharothrix, Brevibacterium, Microbacterium genus and Bacillus, B.
megaterium were also isolated in Paleolithic cave paintings in southern Spain [22–24].
Apart from bacteria, wall paintings also contained fungi from the genus of Cladosporium,
Penicillium, Nectria and yeast from the genus of Rhodotorula [25].
Okunye et al. [26] studied species of fungi capable of causing the degradation of paint. Thirty
six samples obtained from various locations in Southeastern Nigeria helped isolate Rhizopus
and Aspergillus spp., which were the most frequently present. Other genuses of fungi as: Absidia,
Monilia, Alternaria, Fusarium and Penicillium were also detected. [26]. Aspergillus, Penicillium,
Botrytis, Acrodictys, Mucor, Absidia, Atternaria, Cladosporium, Rhizopus, Cephalosporium, Fusarium,
Helminthosporium, Trichocterma, Acremonium, Pullularia, Monilia, Epicoccum and Udeniomyces
were likewise isolated genuses from the painted surfaces investigated in studies [27–29]. Likewise,
species of fungi Aspergillus niger and Cladosporium sp. were isolated from paint coatings [30, 31].
Fungi such as Aureobasidium sp. and Cladosporium sp. [9, 11, 14, 15, 32–36] were also frequently
found. In addition, Lugauskas et al. (2003) identified fungi from the genera of Aureobasidium
pullulans, Aspergillus niger, Aspergillus versicolor, Cladosporium cladosporioides, Paecilomyces
sulphurellus, Trichoderma viride, Ulocladium atrum as capable of degrading polymer materials [37].
The following fungi species: Acremonium charticola, A. strictum, A. kiliense, Acremonium
spp., Aspergillus sydowii, Aureobasidium pullulans, Beauveria sp., Cladosporium sp., Cladosporium
sphaerospermum, Chrysosporium sp., Engyodontium album, Mycelia sterilia, Scopulariopsis
brevicaulus, Verticillium lecanii, Verticillium suchlasporium, Verticillium sp. were isolated from
walls painted in antiquity [38]. The same species were isolated and described in study [39],
which also isolated species from the genera of Arthrobacter and Bacillus [39]. Researchers
studying similar subjects isolated the following genera of fungi and bacteria: Pseudomonas
alcaligenes, Nocardia asteroides, Arthrobacter spp., Bacillus subtilis, Pseudomonas aeruginosa,
Streptococcus spp., Pseudomonas fluorescens, Micrococcus roseus, Bacillus pumilus, Streptomyces
spp., Micrococcus luteus., Alternaria alternata, Aspergillus versicolor, Chaetomium globosum,
Cladosporium cladosporoides, Aspergillus terreus, Aspergillus oryzae, Aureobasidium spp.,
Penicillium spp. and Penicillium stoloniferum [40].
Studies carried out by Romero-Noguera et al [41, 42] looked into the biodeterioration
of turpentine varnishes. Based on their findings, researchers identified the following
microorganisms capable of degrading Venetian turpentine coatings: Chrysonilia sitophila,
Streptomyces celluloflavus, Bacillus amyloliquefaciens, and Arthrobacter oxydans [41]. Moreover
they identified microorganisms capable of degrading Sandarac coatings (Chrysonilia
sitophila, Penicillium chrysogenum, Rhizopus oryzae, Mucor indicus) and Manila Copal coatings
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(Aspergillus niger, Aureobasidium pullulans, Chrysonilia sitophila, Penicillium chrysogenum,
Pleospora herbarum, Rhizopus oryzae) [42].
Polish studies also looked into the presence of microorganisms in paint coatings. Scientists
isolated fungi present in paint inside residential buildings and public utility buildings. The following
genera of fungi were isolated in residential buildings: Acremonium stictum, Alternaria alternata,
Alternaria tenussima, Aspergillus niger, Aspergillus versicolor, Penicillium cyclopium, Penicillium
janthinellum, Penicillium thomii, Penicillium viridicatum, Rhizopus nigricans, Trichoderma viride.
Researchers also studied fungi in the Registry Office, an office building, a university, a polytechnic,
a primary school, a hospital, and a hotel in the Silesian Voivodship. 20 species of fungi were
isolated, including: Acremonium stictum, Alternaria alternata, Aspergillus niger, Aspergillus ustus,
Aspergillus versicolor, Cladosporium cladosporides, Chateomium globosum, Gliocladium catenulatum,
Gliocladium roseum, Memnoniella echinata, Paecilomyces variotii, Penicillium albidum, Penicillium
chrysogenum, Penicillium citrinum, Penicillium rugulosum, Penicillium terrestre, Penicillium variabile,
Scopulariopsis brevicaulis, Stachybotrys chartarum, Trichoderma viride [4].
Research was also conducted on fungi present in paint coatings in food industry plants,
breweries, dairies, and fruit preserve factories.
Alternaria sp., Cladosporium herbarum, Leptographium sp,. Phoma violacea, Trichothecium
sp., Trichoderma koningii were isolated in food industry plants, Aspergillus flavus, Botrytis
cinerea, Acremonium sp., Chalaropsis sp. in breweries, and Geotrichum sp. and Fusarium sp. in
dairies. Paint in fruit preserve factories was shown to contain the following species of fungi:
Aspergillus nidulans, Aspergillus flavus, Aspergillus oryzae, Aspergillus versicolor, Penicillium
glabrum, Penicillium italicum, Penicillium spinulosum, Botrytis cinerea, Aureobasidium pullulans,
Cladosporium elatum, Cladosporium herbarum and Alternaria tenuis, as well as various
species of Fusarium sp. In chocolate-processing factories, the following species of fungi were
detected: Mucor hiemalis, M. racemosus, M. spinosus, Rhizopus nigricans, Aspergillus ochraceus,
Aspergillus versicolor, Penicillium cyclopium, Penicillium expansum,, Penicillium globrum and
Penicillium brevicompactum. The following fungi were isolated in a juice factory: Cladosporium
sphaerospermum, Chaetophoma sp. and Acermonium strictum [4].
Table 1. Microorganisms isolated from paint exposed to atmospheric conditions [2, 43]
Oil Emulsion Oil Emulsion
Microorganism Microorganism
paint paint paint paint
Fungi Bacteria
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Oil Emulsion Oil Emulsion
Microorganism Microorganism
paint paint paint paint
Flavobacterium
Cladosporium sp. + + + +
marinum
Pseudomonas
Stemphylium consortiale + + +
maltophilia
Torula nigra +
2.2. Biodegradation
86
In addition, the authors of the study determined bacterial generation time under growth
conditions. These were, respectively, 44.67 minutes for Pseudomonas flourescens; 45.04
minutes for Bacillus subtilis; 35 minutes for Bacillus licheniformis, 18 minutes for Bacillus cereus,
19 minutes for Bacillus megaterium, and 53 minutes for mixed bacterial cultures.
Studies were also conducted to investigate the biodegradation of waste produced in the
process of varnish removal. Arquiaga et al. [52] studied the decomposition of paint stripping
wastewater, which contained methylene chloride at the concentration of 5000 mg∙dm-3,
phenol at the concentration of 1800 mg/dm3, and other organic ingredients, such as paraffin
waxes, cellulose derivatives, petroleum sulfonates, and naphthalene at the total concentration
of 2200 mg∙dm-3. Active sludge was used. The study identified bacteria present in active
sludge and their ability to grow in the presence of phenol and varnish stripping wastewater.
In addition, researchers investigated whether these substances can serve as a source of
carbon for the bacteria [52]. They isolated such bacteria as Enterobacter sp., Klebsiella sp.,
Corynebacterium sp., Pseudomonas sp., Bacillus sp., Acinetobacter sp.. The bacteria most
frequently isolated from the wastewater were Gram-negative, with the majority belonging to
the Pseudomonas genus. Gram-positive bacteria, represented by the genus Bacillus, were less
87
numerous. Studies showed that most of the isolated bacteria can biodegrade phenol and paint
stripping wastewater. However, the most important role in the process is played by bacteria
from the Pseudomonas and Bacillus genus [52].
Vanderberg-Twary et al. [53] researched the biodegradation of solvents used in varnish
removal agents, focusing on the biodegradation of typical agents of this kind. They prepared
two kinds of agents: one contained toluene, acetone, and dichloromethane, the other
– dichloromethane, isopropyl alcohol, and methanol. The study used two types of bacteria:
Hyphomicrobium sp. and Rhodococcus rhodochrous sp. [53].
The study showed that the selected strains of bacteria are able to degrade solvents present
in paint stripping agents [53].
In patent US8202424, Almadidy et al. [54] described the biodegradation process of paint
stripping waste containing high levels of organic substances. Their study used two kinds of
bacteria: Pseudomonas fluorescens and Bacillus subtilis and one type of fungus, Cunningham
elegans. They tested the changes in the COD of waste samples subjected to microbial activity.
The study showed that it is possible to achieve as much as 47% decrease in COD with the use
of the selected bacterial strains [54].
4. Conclusions
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