By Zaharaddeen, MA; Owuna, JE; Yahaya, I; Nuhu, KU; Abdumalik, AA; Adam, IM; Orole, OO (2023). Greener Journal of Epidemiology and Public Health, 11(1): 35-40.

Greener Journal of Epidemiology and Public Health ISSN: 2354-2381 Vol. 11(1), pp. 35-40, 2023 Copyright ©2023, Creative Commons Attribution 4.0 International. https://gjournals.org/GJEPH

Molecular Characterization of Bacillus Species Associated with Deterioration of Painted Surfaces in Nasarawa State University, Keffi

Zaharaddeen, MA^1*; Owuna, JE²; Yahaya, I²; Nuhu, KU³; Abdumalik, AA²; Adam, IM², and Orole, OO⁴

1. National Agency for Science and Engineering Infrastructure, NASENI, Garki, Idu Industrial Area, Abuja, Nigeria.

2. Department of Microbiology, Nasarawa state University Keffi, Nigeria.

3. Department of Animal Science, College of Agriculture, Science and Technology, Lafia.

4. Department of Microbiology, Federal University of Lafia, Nigeria.

ARTICLE INFO	ABSTRACT
Article No.: 072323068 Type: Research Full Text: PDF, HTML, PHP, EPUB, MP3	Biodegradation of painted walls involves the disintegration of painted layers by microorganisms. The aim of this study was to isolate and characterize Bacillus species associated with deterioration of paint surfaces. This study was carried out at the Nasarawa State University Keffi, Nasarawa State, Nigeria. Conventional methods based on biochemical and morphological characteristics were used to isolate Bacillus species and the confirmation was done employing the 16S rRNA gene sequencing. The total viable counts were lowest in the Faculty of Natural Sciences (1.04 x 105cfu/g) and highest in the Faculty of Social Sciences with a count of 2.0 x105cfu/g. The incidence of Bacillus species isolated showed that B. subtilis 7(28%) was the most predominant species, while B. pumilus with only one isolate (4%) was the least isolated. Painted walls from the Faculty of Social Science were the least colonized by the Bacillus species (12%) and walls painted from the Faculty of Natural Sciences and Administrative Block had higher Bacillus colonization rates of 24% and 16% respectively. The isolated bacterial species are capable of defacing painted walls; hence it is recommended that building owners should routinely repaint their wall surfaces.
Accepted: 23/07/2023 Published: 31/07/2023
*Corresponding Author Zaharaddeen, M.A. E-mail: zahramad001@ gmail.com
Keywords: Deterioration, Bacillus, Paints, Colonization, wall.

INTRODUCTION

Microbial interaction and association have the potential to cause various physical and chemical processes that result in destruction. Like any other material, buildings are susceptible to the colonization, deterioration, and degradation caused by microorganisms, also known as “weathering”. Architectural structures, such as buildings and bridges, that come into contact with water, soil, waste, sewage, plant materials, or any organic matter can experience deterioration. Despite their strong and solid nature, these structures are subject to the slow and inevitable process of corrosion after microbial colonization, given the right conditions (Braums, 2002). The primary purposes of paint are to enhance and protect the surface. Paint is applied to safeguard the surface from erosion, and oxidation, and for decorative purposes (Stefanie et al., 2009). The presence of utilizable substances in building materials makes buildings prone to microbial attack and colonization. Materials such as pigment, thinners, binders and driers which are the main components of paints are metabolized by microorganisms for the generation of energy (Pandey and Kiran, 2020; Parker, 2000). The painted surfaces contain ample nutrients and create a microenvironment for prior colonization before access to the building by the microorganisms. During this invasion, microorganisms synthesize corrosive acids which can solubilize the lattice structure of walls.

The biodegradation of painted walls involves the breakdown of paint layers by microorganisms into physical signs of decay and changes in such buildings. Heterotrophic bacteria metabolize organic compounds in the paints as growth substrates to produce acids that cause the paint to discolour or change its consistency, especially in tropical climates where environmental conditions are favourable for the growth and formation of spores. The colonization rate, microbial type and population are dependent on the substrate composition in the paint and environmental conditions (humidity, temperature, light and possibly pH) in which the building is situated (Ciferri, 1999). The deterioration of painted walls is a dissimilatory process encompassing the microbial release of wastes and/or other metabolites into the microenvironment and the subsequent reaction with the components of the painted wall, which ultimately alters its aesthetic value and reduces its shelf-life (Yamuna et al., 2021). In cases where the biodegradation process is not visible to the naked eye, the internal structure of the underlying substrate is still affected (Ranalli et al., 2009). Phototrophic microbes were previously reported as the main colonizers, and when they die off, the cell wall is lysed to provide nutrients that promote fungal growth (Gaylarde and Gaylarde, 2000). Microorganisms that form specific communities interact in different ways with mineral materials and their external environment. This complex phenomenon occurs simultaneously with many physical and chemical decomposition processes. The main groups of microorganisms involved in the deterioration of painted walls are bacteria, fungi, and lichens that can grow on applied paint films as well as in solvent and water-based coatings (Gaylarde and Gaylarde 2005; Yamuna et al., 2021).

Paint contains harmful chemicals volatile organic compounds that can become airborne and cause serious environmental and health problems. Paints contain one or more pigments of different chemical composition, high levels of mercury (Yamuna et al., 2021; Sharma, 2006), which can enter the body through inhalation or ingestion. It can cause nerve, liver and kidney damage, along other symptoms. Many paint components with their additives (adhesives, surfactants, thickeners, etc.) are degradable despite ensuring elegant quality of the product (Ogbulie and Obiajuru, 2004). When colors start to fade, the most obvious cause is the low quality of such a paint that leads to premature failure of the color system. Mold and mildew are black, bluish, translucent structures whose growth is encouraged by excessive humidity. While these organisms are detrimental to health, they also compromise the basic structural integrity of buildings. Microorganisms damage coatings on painted surfaces, degrading them by causing discoloration, increasing porosity of the coating, reducing physical strength and shelf life, and making the surface more susceptible to moisture penetration (Okunye et al., 2013; Maduka, 2019). Common biological agents causing deterioration in construction works include Nitrifying bacteria, Cyanobacteria, Thiobacteria, and fungal genera Aspergillus, Fusarium, Penicillium, Alternaria, Trichophyton and Cladosporium (Videla and Herrerii, 2005). These agents cause damage when environmental parameters such as water, humidity, temperature, UV light and ventilation are optimal (Singh, 2004).

While painted walls with defaced discoloration are common in Keffi, Nasarawa State, Nigeria, contaminating and associated bacteria species has not been carefully studied in North central Nigeria where environmental factors favouring their optimal growth and development are prevalent. The study isolated and characterized Bacillus species associated with the deterioration of paint surfaces in Nasarawa State University, Keffi.

 

MATERIALS AND METHODS

Study Area and Sampling

The study was carried out at Nasarawa State University Keffi. Keffi is geographically located in the North-central zone of Nigeria between latitude 8^o 50^o North and longitude 7^o 52^o East. Five samples were collected each by swab sampling from the Faculty of Natural and Applied Sciences, Administration Block, Convocational Square, Faculty of Social Science and Faculty of Art on the University campus. Samples were collected by swabbing from deteriorating painted surfaces after scrapping to a depth of 3–10 mm from dried areas with extensive peelings and visible discolouration.

Isolation and Identification of Bacillus Species

The collected samples were processed according to the methods of Omar et al. (2013) by dissolving 1 g into 50 mL distilled water and shaking at 200 rpm at 37^oC for 30 min, after which approximately 0.1 mL of the suspension was inoculated and spread on a Nutrient agar plate using the spread plate technique. The cultured plates were incubated at 37^oC for 24 h after which discrete colonies were transferred onto Mineral Salt Medium (MSM) at 37^oC for 72 h for the isolation of Bacillus species. The morphological characteristics of the isolates were recorded, and the Gram staining reaction, spore staining, catalase, indole, Voges Proskauer, and Methyl Red tests were conducted to identify the bacteria.

Molecular Identification Using 16S rRNA

The molecular identification of the isolated Bacillus species was carried out by partial sequencing of the 16S rRNA gene. The molecular identification of the isolates was performed by amplification using universal primers, 27F (5′-AGAGTTTGATCMTGGCTCAG-3′) and 1492R (5′- TACGGYTACCTTGTTACGACTT-3′). The amplified gene products were purified and sequences were obtained through ABI PRISM Big DyeTM Terminator Cycle Sequencing Ready Reaction Kit (PE Biosystem, USA) by using universal sequencing primers, i.e, 518F (5′-CCAGCAGCCGCGGTA TACG-3′) and 800R(5′-TACCAGGGTATCTAATCC-3′).

 

RESULTS

The total bacterial counts from the deteriorated paint surfaces of Nasarawa State University, Keffi was 1.04 x 10⁵ cfu/g from the Faculty of Natural and Applied Science (FNAS) and Convocation Square (CONVOC) as the lowest respectively, while the Faculty of Social Sciences with 2.1 x 10⁵ cfu/g was the highest (Figure 1).

Figure 1: Total Viable Counts of Bacteria isolated from deteriorated paint surfaces of Nasarawa State University, Keffi

Key: FNAS – Faculty of Natural and Applied Science, ADMIN – Administration Block, CONVOC – Convocational Square, F.SOS – Faculty of Social Science, F.ART – Faculty of Art

Based on 16S rRNA sequence homology, all the four isolates showed more than 99 % similarity with Bacillus species isolates available in National Centre for Biotechnology Information (NCBI) GenBank library. Thus, the isolates were confirmed as the strains of B. thuringiensis B. subtilis, B. cereus and B. pumilus as shown in Fig. 2. The nucleotide sequences of each isolate was submitted to NCBI GenBank.

Figure 2: Agarose gel electrophoresis of the 16SrRNA gene of bacteria isolates. L represents the molecular ladder, Lane 1 = Bacillus thuringiensis strain, Lane 2 = Bacillus subtillis strain, Lane 3 = Bacillus cereus, Lane 4 = Bacillus pumilus strain.

The occurrence of Bacillus species isolated from deteriorated paint surfaces of Nasarawa State University, Keffi in Table 1 showed that all the Bacillus species (Bacillus thuringiensis, Bacillus subtillis, Bacillus cereus and Bacillus pumilus were isolated from the FNAS, while Bacillus pumilus was not isolated from any other location where samples were collected. The Convocational Square and Faculty of Art both had Bacillus subtillis (5.9%) and Bacillus cereus (5.9%) each.

Table 1: Occurrence of bacteria isolated from deteriorated paint surfaces of Nasarawa State University Keffi

Sampling Location	Sample size	Bacillus thuringiensis	Bacillus subtillis	Bacillus cereus	Bacillus pumilus
FNAS	5	2 (8%)	1 (4%)	2 (8%)	1 (4%)
ADMIN	5	1 (4%)	2 (8%)	1 (4%)	0
CONVOC	5	0	1 (4%)	1 (4%)	0
F.SOS	5	1 (4%)	2	0	0
F.ART	5	0	1 (4%)	1 (4%)	0
Total	25	4 (16%)	7 (28%)	5 (20%)	1 (4%)

Key: FNAS – Faculty of Natural and Applied Science, ADMIN – Administration Block, CONVOC – Convocational Square, F.SOS – Faculty of Social Science, F.ART – Faculty of Art

 

DISCUSSION

Metabolism by microorganisms and the release of secondary metabolites is a major route for the degradation of paint components. Deteriorating painted building surfaces is home to a number of bacterial species. Four different species of Bacillus strains were isolated from the painted walls in the Nasarawa State University Keffi in the study. The present study agrees with the finding by Poyatos-Jiménez et al. (2021) who isolated Bacillus species from coated Outdoor Canvas Paintings. The authors isolated all the Bacillus species obtained in this study except Bacillus licheniformis. Other authors like Shinkafi and Haruna (2013) in Sokoto State, Nigeria and Ogbulie and Obiajuru (2004) in Owerri, Imo State identified Bacillus strains different from the species isolated in the present study.

Bacillus are spores’ formers, an advantage that allow them survive unfavorable environmental conditions prevailing on the walls from one season to another. High amount of substrate in the paints and water activity within the wall encourages bacterial growth and biofilm formation which promotes further colonization by other bacterial and fungal genera. Spore formers like Bacillus species maintain viability employing β-glucosidase enzymatic activity. The enzyme is involved in the hydrolysis of cellulose, a component present in the paints. The isolated bacteria in the study employed such mechanisms as osmolytes production, which maintains cell integrity and water uptake from the air (Cetiner et al. 2017), and ability to survive limited organic matter stress prevalent in paint (Ma et al. 2015).

Colouration of the walls was due to fungal hyphae penetrations and enzymes release into building paints. High microbial population and pigments release into the surrounding environment as a result of metabolic activities of colonizers has also been attributed to cause discolorations on the painted walls (Abdel-Haliem et al., 2013). The discolouration produced is also dependent on bacteria and type of paint involved (Ashwini and Anchana, 2018). The presence of Bacillus species probably originated from the environment (Anele et al., 2019). Though Bacillus species are not considered human pathogens, their presence is a sign of the unhygienic nature of the paints that encourages colonization and contamination.

 

CONCLUSION

The four Bacillus species isolated from deteriorating painted wall surfaces of Nasarawa State University Keffi are known to originate from the environment. Though these bacterial species are not recognized as pathogens, they are spore formers that can survive extended time and adverse environmental conditions. They produce enzymes that can hydrolyse and destroy the matrix structure of paints and walls. Further research is proposed to determine the effects of produced metabolites and the waste products formed.

 

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Cite this Article: Zaharaddeen, MA; Owuna, JE; Yahaya, I; Nuhu, KU; Abdumalik, AA; Adam, IM; Orole, OO (2023). Molecular Characterization of Bacillus Species Associated with Deterioration of Painted Surfaces in Nasarawa State University, Keffi. Greener Journal of Epidemiology and Public Health, 11(1): 35-40.

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