bacterial and fungal biodeterioration of discolored building

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World Journal of Microbiology andBiotechnology ISSN 0959-3993Volume 33Number 11 World J Microbiol Biotechnol (2017)33:1-9DOI 10.1007/s11274-017-2362-y

Bacterial and fungal biodeterioration ofdiscolored building paints in Lagos, Nigeria

Olayide Obidi & Foluke Okekunjo

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World J Microbiol Biotechnol (2017) 33:196 DOI 10.1007/s11274-017-2362-y

ORIGINAL PAPER

Bacterial and fungal biodeterioration of discolored building paints in Lagos, Nigeria

Olayide Obidi1 · Foluke Okekunjo1

Received: 1 June 2017 / Accepted: 27 September 2017 © Springer Science+Business Media B.V. 2017

Introduction

Discolorations on a wall painting typically destroys its aesthetics, stability and original intent of beautifying and decorating the environment. The constituents of wall paint-ings including the pigment, binder, thinner and drier are all susceptible to microbial attack (Pelczar et al. 2005). This is because they provide carbon sources and nutrients to facili-tate proliferation of practically all species of microorgan-isms on a painted wall. Although, discoloration has been implicated generally to the presence of moisture and other atmospheric pollution, it is important to note that the coloni-zation by microorganisms as well as their metabolic products form biofilms resulting in unacceptable appearance, aes-thetic biodeterioration and degradation of a painted layer. Biochemical deterioration on painted walls in most cases is a dissimilatory process whereby the live organisms excrete waste products or other substances that react chemically with the painted wall component, thus compromising its aesthetic value and deteriorating its properties (Hochmannova and Vytrasova 2010). In cases where biodeterioration is not vis-ible to the naked eye, it still affects the internal structure of the underlying substrate (Ranalli et al. 2009). Previous studies carried out in Latin America have reported phototro-phic microorganisms as the primary colonizers, whose death and lysed cell walls supply organic matter that promote the growth of fungi (Gaylarde and Gaylarde 2000). Microor-ganisms form specific communities which interact in many different ways with mineral materials and their external envi-ronment. This complex phenomenon occurs synergistically with many physical and chemical degradative processes. The major groups of microorganisms involved in painted wall deterioration are bacteria, fungi and lichens which can grow on applied paint films as well as in solvent and water-based coatings (Gaylarde and Gaylarde 2005; Stefanie et al. 2009).

Abstract Microbial induced discolorations are an unsightly feature occurring on painted walls in Lagos, the commercial hub of Nigeria. Very few studies have been carried out conventionally about the microbial community structure of discolored painted walls in Nigeria therefore, knowledge of the true microbial diversity is elusive. To fur-ther our understanding of the phylogenetic diversity of rep-resentative microbial community on 40 discolored and three clean-looking buildings, a comparative DNA sequence anal-ysis of 16S rDNA genes was undertaken. Following DNA extraction, portions of the rDNA genes were amplified by PCR and sequenced. Resulting sequences were compared with GenBank data base sequences. Fifteen unique fungal sequences and one bacterial sequence were obtained. Major-ity (37.50%) of rDNA sequences analyzed, represent the genus Meyerozyma of which the novel fungus Meyerozyma guilliermondii, which to our knowledge, has not yet been implicated in painted walls was detected. Clones from the discolored painted wall isolates also produced a data set in which 31.25% of sequences were related to Fusarium pro-liferatum and 6.25% were Pseudomonas aeruginosa. The remaining sequences clustered with members of the gen-era Candida (6.25%), Aspergillus (12.5%) and Cerrena (6.25%). The study provides reliable data on microbial com-munities on painted walls and information for paint biocide formulation.

Keywords Discoloration · Building paints · Microbial community · Biocide

* Olayide Obidi [email protected]

1 Department of Microbiology, University of Lagos, Lagos, Nigeria

Author's personal copy

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World J Microbiol Biotechnol (2017) 33:196

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Biodegradation of painted walls involves the disintegration of painted layers by the microorganisms. This form of bio-deterioration is mostly visual and causes physical changes on the buildings such as discoloration through microbial pigments and excretion of other metabolic products. Het-erotrophic bacteria can use organic compounds from the paint layer as growth substrates, producing acids, which cause discoloration of the paint or change its consistency especially in tropical climates, where prevailing heat and humidity conditions favor growth and sporulation. Humidity levels in the range of 75–98% has been observed to result in the germination of spores to mycelium. Furthermore, time required for developing hyphae to produce further spores reduces from 11 to 8 h when such high humidity is maintained with aeration (Hassouni et al. 2007). Elevated airborne propagule concentrations thus, increase the risk of biodeterioration, particularly in tropical environments (Borrego et al. 2010). The rate of microbial colonization, types of microorganisms and population can be a function of the substrate composition and environmental conditions (humidity, temperature, light and probably pH etc.) (Ciferri 1999). Ascomycetes such as Chaetomium may degrade vari-ous polymers and have been found growing on cellulose-containing substrates (Abín et al. 2002; Portugal et al. 2009). Fungi can prompt physical biodeterioration by mycelial growth, hyphal penetration and chemical biodeterioration of the substrate through their metabolic products (includ-ing organic acids, mycotoxins and pigments) and enzymatic activities resulting in detachment of fragments (Ravikumar et al. 2012). Saprophytic fungi can colonize a wide range of substrates and ecological niches including painted walls. The extent of such colonization depends on the genetic poten-tial of the species involved and on prevailing environmental conditions (Nugari and Roccardi 2001). Pigment producing microorganisms release different colored stains and patches on painted walls as a consequence of their metabolic activi-ties which, once formed are almost impossible to remove (Ravikumar et al. 2012). Discoloration is usually the most common manifestation of fungal contamination and an indi-cation of eventual structural damage due to the consider-able capacity of many fungi to degrade a whole range of organic compounds (Florian 2003). Advancement in science has shown that conventional biochemical identification will not give a complete picture of total microorganisms present in environmental samples (Ward et al. 1990). Furthermore, microorganisms present on deteriorating painted buildings have not been fully characterized in Nigeria. It is in this con-text that the present study sets out to evaluate bacteria and fungi occurring on discolored painted buildings using DNA identification methods in order to capture important micro-organisms responsible for biodegradation and subsequent discoloration of painted buildings. This paper reports one of the very few studies focusing on the identity of culprits

in microbial-induced discolorations, thereby providing new and interesting information for biocide formulators to target specific organisms. It underscores the novelty and aim of the present work and its potential impact on paint formulation to prevent further discolorations and improve building aes-thetics especially in Lagos, the commercial hub of Nigeria.

Materials and methods

Site description and sampling

Samples for investigation of microbial growth were col-lected in duplicates by swab-sampling from 40 randomly selected discolored painted buildings from eight local gov-ernment areas (LGAs) of Lagos state, Nigeria. The LGAs include: Badagry, Shomolu, Epe, Amuwo Odofin, Mushin, Surulere, Ojo and Ifelodun LGAs. The buildings (Fig. 1, 2) were analyzed between June and August 2016 (a rainy season in Nigeria) by sampling the exterior, visually dis-colored areas exposed to minimal sunlight with moderate temperature of ~ 26 °C, average rainfall of ~ 186.8 mm and humidity of ~ 86%. The sterile swab sticks were applied on the external surface layers of the painted walls that contain discolorations and presumed biological growth as described by Gorbushina et al. (2004). The inoculated swab sticks were subsequently taken to the laboratory for culturing. Similarly, control samples were obtained from three randomly selected clean-looking painted walls show-ing no visible discolorations from Amuwo Odofin, Ojo and Badagry LGAs.

Cultivation of bacteria and fungi

Each inoculated swab stick was suspended in labeled plain bottles containing 4 ml of peptone water (pH 7.0) to dislodge microbial cells from the swab sticks. Subsequently, aliquot (0.1 ml) of the suspension was inoculated on respective media. Heterotrophic bacteria were isolated on nutrient agar and moulds on potato dextrose agar. Sterile spreader was used to spread the inoculum entirely on the plates. The inoc-ulated plates were incubated aerobically at 37 °C for 24 h for bacteria and at room temperature (27 ± 2 °C) for 72 h for fungi. At the end of the incubation period, well defined, pigmented colonies that developed were counted, routinely sub-cultured on fresh plates to purify and identified. Total viable microbial cell number was calculated and expressed as colony forming units per gram (CFU g− 1) (Nwachukwu and Akpata 2003). The cultures were maintained on fresh agar medium and stored at 4 °C for further use. All tests were carried out in duplicates.



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Identification of bacteria and fungi

Morphological and microscopic observation

Morphological studies were carried out to determine col-ony size, shape, elevation and pigmentation (Cheesbrough 2008). Pigment production was established by observation of coloration on nutrient agar and potato dextrose agar after aerobic incubation at 37 °C for 24 h and at room temperature (27 ± 2 °C) for 72 h for bacteria and fungi respectively (Rojas et al. 2012). Microscopic observations to determine microbial viability were made using optical microscope.

Biochemical identification tests

Pure representative, morphologically different bacterial colonies obtained from the incubated plates were initially evaluated by conventional tests such as Gram stain, cata-lase, oxidase, motility, indole production, MRVP, oxi-dative fermentative, carbohydrate utilization and urease activity. Additional tests included nitrate reduction, citrate utilization, H2S production, hydrolysis of starch, and spore detection test (Cheesbrough 2008). Purified fungal iso-lates were subsequently examined by a light microscope as described by Harrigan and Mc Cance (1976). Thus, all the

Fig. 1 Representative dis-colored painted walls from a Amuwo Odofin, b Ojo, c Badagry, d Epe, e Ifelodun, f Shomolu, g Mushin and h Surulere LGAs

Fig. 2 Representative clean-looking painted walls from a Amuwo odofin, b Ojo and c Badagry LGAs



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isolates were identified to the genus level based on their morphology and sporing structures.

DNA extraction and 16S rDNA PCR

Representatives of the major clusters were analyzed by 16S rDNA sequence analysis as described below. The genomic DNA of the isolates was extracted using Zymo Research (ZR) Fungal/Bacterial DNA MINIPREP™ KIT (Zymo Research Corp, USA) according to the manufac-turer’s recommendations. The pure bacterial isolates were sub-cultured into nutrient broth while the fungi were sub-cultured into Sabouraud Dextrose Broth. 16S rDNA was amplified with primers 27F (5′-AGA GTT TGATCMTGG CTC AG-3′) and 1525R (5′-AAG GAG GTGWTCCARC CGCA-3′) (Ettenauer et al. 2010). The Internal Tran-scribed Spacer (ITS) primers used for fungal amplifica-tion consisted of ITS-1 (5′-TCC GTA GGT GAA CCT GCG G-3′) and ITS-4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) (White et al. 1990). PCR cocktail mixture (New Eng-land Biolabs™, UK) was prepared as previously reported (Blaiotta et al. 2002), while PCR conditions consisted of 36 cycles (5 min at 94 °C, 30 s at 94 °C, 3 min at 56 °C, 45 s at 72 °C, 7 min at 72 °C) plus one additional cycle at 10 °C as a final chain elongation. The amplicons from the reaction were loaded on 1.5% agarose gel to verify the presence of the PCR products using 1 kb plus lad-der (Thermo Scientific) with Gene Amp® PCR System 9700 (Applied Biosystems™, USA) thermocycler at 100 V for 40 min, purified by adding the Exo/SAP master mix involving 50.0 µl Exonuclease I (NEB M0293) 20U µl−1 and 200.0 µl Shrimp Alkaline Phosphatase (NEB M0371) 1U µl−1 to a 0.6 ml micro-centrifuge tube and sequenced.

DNA sequencing and phylogenetic analyses

The DNA sequences were determined by the termination method using the BigDye™ Terminator v3.1 Cycle Sequenc-ing Kit (Applied Biosystems™) according to the manufac-turer’s instructions. The sequences were analyzed by the ABI3500XL genetic analyzers (Applied biosystems™) with a 50 cm array, using POP7. Comparison for DNA similar-ity was carried out using the GenBank and EMBL database (http://www.ncbi.nlm.nih.gov/Blast.cgi) (Altschul et al. 1997). Subsequently, phylogenetic analysis was performed using Molecular Evolutionary Genetics Analysis version 7.0 (MEGA 7.0) for bigger datasets (Tamura et al. 2004; Kumar et al. 2016) after multiple alignment of data by Clusta l W1.8 (Thompson et al. 1994). Distance matrix and neighbor-join-ing methods (Saitou and Nei 1987) were applied for tree construction.

Results

Culture-dependent methods as well as conventional and molecular technique of microbial identification, were used to obtain bacterial and fungal strains for laboratory experi-ments (Ranalli et al. 2005; Alakomi et al. 2006). In this study, a bacterial strain belonging to the genera Pseudono-monas and seven fungal strains belonging to the genera: Meyerozyma, Aspergillus, Fusarium, Cerrena and Candida were found to occur repeatedly in the discolored painted buildings while only Fusarium and Aspergillus niger at minimal population density of 0.2–0.4 × 102 CFU g− 1 were found in the clean-looking painted buildings which served as controls. The bacterial and fungal species composition from different LGAs are presented in Table 1. All iso-lated strains produced distinct pigmentation with reason-able color yields such as cream, white, bluish-black, light brownish, grayish-brown and brownish-black which were visually observed (Table 1). The results obtained in the pre-sent study showed that pigmented heterotrophic fungi were prevalent in all the discolored painted walls monitored hav-ing a population density ranging from 0.02 × 102 to 3.0 × 102 CFU g− 1. Pseudomonas aeruginosa was also detected in four of the discolored painted walls but only in Epe LGA with population density ranging from 0.2 × 103 to 3.3 × 103 CFU g− 1. In this study, fungi belonging to the genera Asper-gillus were isolated from 20 painted walls with population density ranging from 0.02 to 3.0 × 102 CFU g− 1. Other iso-lated strains on the discolored painted walls included Cer-rena sp. (NIOCC 2a) which occurred at 0.4 × 102 CFU g− 1 and Candida tropicalis which was observed in two of the painted walls samples at a population density of 1. 2 × 102 CFU g− 1. Meyerozyma guilliermondii was isolated in three of the discolored painted walls in this study at population density ranging from 0.4 × 102 to 0.6 × 102 CFU g− 1. Fusar-ium proliferatum was observed both in discolored and clean looking painted walls at population density of 0.2–0.6 × 102 and 0.2 × 102 CFU g− 1 respectively.

Discussion

This paper describes a novel study into the identification of the diversity of microbes in microbial-induced discolored buildings in Lagos, Nigeria using 16S rDNA analysis. Such studies have not been reported thus far in Nigeria, and this may be the first one that reports such investigations. In this study, the mechanisms and biochemical processes involved in the biodeterioration include biological, mechanical and chemical interactions between isolated microbes and the paint components. It is concluded that biodeterioration was triggered when microorganisms colonize clean-look-ing building paints, remain viable and proliferate. This


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proliferation leads to obvious deterioration over time as observed in this study. Evaluation of the strains isolated from various LGAs confirmed the considerable viability of the strains and their potential for spore production as a survival strategy (Olutiola and Nwaogwugwu 1982; Shi-rakawa et al. 2002; Al-Juboory and Juber 2013). The gen-era detected (Pseudonomonas, Meyerozyma, Aspergillus,

Fusarium, Cerrena and Candida) all possess capacity for spore production (Shirakawa et al. 2002). This spore produc-tion mechanism has been implicated in biodeterioration of substrates such as painted surfaces (McGrath et al. 1999). Pseudomonas aeruginosa detected on discolored building paints have been reported to have other mechanisms to resist desiccation, such as the production of osmolytes which help

Table 1 Bacterial and fungal species composition and groupings from different LGAs in Lagos, Nigeria based on 16S rDNA sequence analysis

a Identification of isolates from different LGAs based on 16SrDNA sequence analysisb Colonial morphology of identified strainsc Microscopic characteristics of isolated strainsd Microbial population density obtained for each isolatee Closest relative based on GenBank comparisonf Accession number (submitted to genbank) and length of base pair of sequence analyzed

Identificationa Colonial morphologyb Microscopic characteristicsc

Population density (CFU g− 1)d

Closest relative by data base entrye

Sequencef

LGA 1 (Epe) P. aeruginosa Cream, circular, entire,

mucoidGram-negative 0.2–3.3 × 103 P. putida ATCC12633 KY511067 (1492)

LGA 2 (Mushin) M. guilliermondii Bluish black with yellow

undersideSeptate hyphae and

conidia0.4–0.6 × 102 M. caribbica CBS5674 LT615287.1 (561)

LGA 3 (Surulere) A. aculeatus Light brownish at center

and whitish at edgesSeptate hyphae, short

conidia in chain on sterigmata

0.02–2.0 × 102 Aspergillus sp. SL2 EU833205.1 (600)

LGA 4(AmuwoOdofin) F. proliferatum 2705 Fluffy, filamentous,

whiteAerial mycelium, micro

and macro conidia Roundish conidi-ophores

0.2–0.8 × 102 F. proliferatum 2487 EU272509.1 (563) F. proliferatum 2705 0.2–0.8 × 102 F. proliferatum 2487 EU272509.1 (563)

Aspergillus sp. SL2 Brownish black with yel-low underside

Primary and secondary sterigmata

0.2–3.0 × 102 A. aculeatus A1.9 KC178662.1 (586)

LGA 5 (Badagry) F. proliferatum 2487 Fluffy, filamentous,


and macro conidia0.2–0.6 × 102 F. proliferatum 2705 EU821492.1 (575)

LGA 6 (Ojo) F. proliferatum 2487 Fluffy, filamentous,


and macro conidia0.2–1.8 × 102 F. proliferatum 2705 EU821492.1 (575)

F. proliferatum 2487 0.2–0.8 × 102 F. proliferatum 2705 EU821492.1 (575)LGA 7 (Ifelodun) Cerrena sp. NIOCC 2a White to grayish brown Thin and thick-walled

hyphae0.4 × 103 F. proliferatum 2705 FJ010208.1 (695)

LGA 8 (Shomolu) M. carribica CBS:5674 Bluish black with yellow

undersideSeptate hyphae with

branched conidi-ophores

0.4 × 102 M. carribica CBS:9966 KY104219.1 (693) M. carribica CBS:9966 0.4 × 102 M. carribica CBS:5674 KY104222.1 (695)

0.4 × 102 M. carribica CBS:5674 KY104222.1 (695) M. carribica CBS:9966 0.4 × 102 M. carribica CBS:5674 KY104222.1 (695) M. carribica CBS:9966 0.4 × 102 M. carribica CBS:5674 KY104222.1 (695) M. carribica CBS:9966 C. tropicalis UZ31_13 Whitish and spreading Thick non-septate

hyphae with dark sporangiospores

1.2 × 102 M. carribica CBS:9966 KM361510.1 (553)



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prevent damage to cells and aid the uptake of water from the air by the organism in humid climates (Cetiner et al. 2017) like Nigeria. The ability to survive osmotic stress and resist desiccation provides them with a clear advantage on outdoor building paint, which is frequently dry. Studies have shown that microbes on building paints may have the ability to survive under harsh climate conditions using the limited organic matter available on the paint films due to special-ized biochemical functions and capabilities (Ma et al. 2015). Some of these biochemical mechanisms involve the produc-tion of some complex enzymes (Rosado et al. 2013) not tested in this study. Strains isolated in this study have been confirmed to have considerable enzyme production potential (Rojas et al. 2012). Another mechanism of survival on out-door building paints is the thick, melanized fungal cell walls which is used to resist the effect of chemical attack such as biocides (Saarela et al. 2004) applied on paint films. Fun-gal hyphae penetrate deeply into building paints and release extracellular enzymes, resulting in aesthetic deterioration and mechanical disintegration due to material loss, pigment contamination, acid corrosion, and enzymatic degradation (Ettenauer et al. 2010; Sterflinger 2010). The high micro-bial population density (0.02 × 102–3.3 × 103 CFU g− 1) and heavy pigmentation explains the reasons for visual discol-orations observed on the painted walls. This is in line with the work of Tomaselli (2003) who reported that most fungi are pigmented and therefore, mostly involved in discolora-tions. The concentrations of microbes in this study estimated to range from 0.02 × 102 to 3.3 × 103 CFU g− 1 was however much lower than that obtained by Abdel-Haliem et al. (2013) who obtained microbes on discolored walls in Egypt at lev-els up to 105–106 CFU g− 1.

The isolation of Pseudomonas aeruginosa in this study corroborates earlier works where species belonging to the genus Pseudomonas have been reported as the first predomi-nant colonizers in a succession of growth on painted surfaces where the polysaccharides of their capsules provide nutrients for growth of other fungi (Ciferri 1999; Morton and Surman 1994). Other studies in agreement also reported that bacteria are generally, the first colonizers on most painted surfaces and provide initial supply of organic matter (Garg et al. 1995; Karpovich-Tate and Rebrikova 1991). Our results agree with earlier works that showed that a large variety of heterotrophic bacteria including Pseudomonas aeruginosa are a common feature on inorganic substrata such as painted walls containing traces of organic material (Bassi et al. 1986; Sorlini et al. 1987; Ciferri 1999; Heyrman et al. 1999; Palla et al. 2002; Tomaselli 2003; De Leo et al. 2012). Members of the Pseudomonas genera have been reported to possess interesting and varied abilities to hydrolyze polymers such as paints and degrade plastics (Cappitelli et al. 2007). Their occurrence as primary colonizers in this study necessitates the development of biocides targeting Pseudomonas species.

Most biocides are generally broad spectrum or multi-tar-geted and not specifically targeting a particular organism. Therefore, in-can biocides and dry film biocides should be formulated to specifically target Pseudomonas species. Recently, a polymer functional silver nanocomposites bio-cide was developed which presented effective antimicro-bial activity against Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli) and Gram-positive bacteria (Staphylococcus aureus and Bacillus amyloliquefaciens) (Guo et al. 2017). Surprisingly, biocides incorporated into paints have proved ineffective as many more microbes develop resistance to antimicrobial agents. Microorganisms have the capacity to adapt rapidly to new environmental conditions and can survive exposure to antimicrobials by using a battery of resistance mechanisms. Microbial resist-ance against different types of biocides has been reported (Morente et al. 2013). While some microorganisms acquire resistance traits from other organisms, many become resist-ant following mutations in the chromosomal gene coding for the target of the growth inhibitory compound (Levy 2002). The pyrithiones are marketed as broad spectrum biocides and are also used in plastics, textiles and dry paint (Arch Chemicals 2013). TPBP (borocide) is an organoborane com-pound mainly used in Japan (Thomas and Langford 2009) where it has been a common antifouling biocide since 1995 (Mochida et al. 2012).

However, Molino and Wetherbee (2008) pointed out that many of today’s antifouling coatings fail to inhibit settling and growth of microalgal biofilms. Therefore, to have a bet-ter understanding and prediction of biocide for optimal paint performance in the real environment, new realistic assays are required to evaluate the efficacy of such biocides. This will aid the environmental risk assessment as well as the development of new biocides.

Currently, the use of nanoparticles is being explored to combat microbes that induce discolorations and aesthetic damage in paints (Obidi et al. 2014). Therefore, emerg-ing nano-structured paints are promising solutions for the final consumer. Mural paintings have been reported to har-bor a wide range of organic and inorganic constituents and thus, provide different ecological niches for a large variety of microbial flora that can exploit and colonize these sur-faces resulting in aesthetic and structural damage (Ciferri 1999). Phototrophs which have bright prospects in areas with abundant sunshine have been isolated from other envi-ronments such as Latin America (Isichei 2009). However, they were not isolated in this study probably because the period of isolation during this study (June–August) portends rainy season in Nigeria which allows some shady scenarios around painted buildings. It is therefore understandable that phototrophs were not isolated due to obvious differences in microclimatic parameters. Claims have been made over many years for a relationship between weather or climate



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and solar variations (Pittock 1978) which contribute to the types of organisms inhabiting different environments. Previ-ous studies have also reported the occurrence of Aspergillus species on painted canvass (Ciferri 1999). The occurrence of Aspergillus sp. could facilitate the discoloration and sub-sequent structural damage of the painted walls by virtue of their mycelial production. Rolleke et al. (1996) and Ciferri (1999) suggested that the hyphae produced by Aspergillus and Penicillium penetrate painted layers resulting in degra-dation of the components such as glues and binders which eventually leads to reduction in its cohesion and subsequent exfoliations, cracking and loss of the paint.

Furthermore, the occurrence of Candida tropicalis in two of the painted walls samples is rather noteworthy. Candida tropicalis has been reported as the most prevalent pathogenic yeast species of the Candida-non-albicans group and has not been implicated in biodegradation of painted walls but rather, in clinical infections (Kothavade et al. 2010). Mey-erozyma guilliermondii, a novel fungus which, to our knowl-edge, has not been implicated in discolored painted walls was isolated in three of the discolored painted walls in this study. Previous studies have reported its occurrence in clini-cal and environmental samples (Savini et al. 2011) as well as in healthy humans (Kam and Xu 2002). In addition, a closely related species Meyerozyma caribbica (anamorph Candida fermentati) (Kurtzman et al. 2011) was also isolated in two of the discolored painted walls in this study. The occur-rence of Cerrena sp. is not unusual being a saprophyte and therefore having the possibility of existing on biodegraded painted buildings. In line with our study, Fusarium prolifera-tum has been previously reported in stone monuments as a major pigment producer involved in discoloration (Savkovic et al. 2016). Generally, rapid progressive discolorations have been observed to modify painted surfaces in places exposed to direct sunlight as well as shady environments. However, color changes in painted surfaces in humid environment (90% RH) has been observed to be more predominant in sun-lit locations than in shady environments (Terrapon and Bearat 2010). The analysis of the 16S rDNA sequences con-firmed that P. aeruginosa CH01 (KY511067.1) was most similar to P. putida ATCC 12633 (AF094736.1) (data not shown) while M. guilliermondii MB14B1 (LT615287.1) was most similar to M. caribbica CBS: 5674 (KY104219.1) and shares closest homology with C. tropicalis UZ31_13 (KM361510.1) (data not shown). The phylogenetic tree (data not shown) also revealed that Meyerozyma, Aspergil-lus, Fusarium have similar intra-species genetic distance of 0.02 between individual isolates as Pseudomonas species indicating similar rate of evolution and the genetic varia-tion between the isolates. The production of pigments by the isolated strains elucidate their direct involvement and roles in discolorations rather than an accidental occurrence as a result of air and wind currents or gravitational settling.

Biocides used in paint industries therefore, should be spe-cific and formulated to target the commonly found organisms on tropical, humid environments.

Conclusion

In this work, we have reported the various microbial strains that contribute markedly to the discoloration and biodeterio-ration of selected painted walls in Lagos, Nigeria. The role of the isolated strains in the observed discoloration which was clearly visible to the naked eye has been facilitated by virtue of their ability to produce pigments. This study identified the microbial communities present on discolored painted buildings and provide data for more detailed study of the ecology and physiology of these group of organisms on painted walls in Lagos, Nigeria.

Acknowledgements This research did not receive any specific Grant from funding agencies in the public, commercial or not-for-profit sectors.

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