MORPHOLOGICAL, MOLECULAR CHARACTERIZATION AND CONTROL OF CYANOBACTERIA FROM WALL PAINTINGS AT NIAH, SARAWAK, TEMBURONG AND PERAK CAVES IN MALAYSIA
DOI:
https://doi.org/10.53555/eijas.v8i2.172Keywords:
Malaysian caves, cyanobacteria, NaOCl, H₂O₂, Ca(OCl)2Abstract
Malaysia is a country of numerous natural wonders, which include a collection of impressive cave formations. Found
all over the country, these caves contain rock art and wall paintings. The samples were collected from wall surfaces
painted caves of Niah, Sarawak, and Tempurung, Perak. Three cyanobacterial genera, have been characterized using a
polyphasic approach by comparing phenotypic characteristics, using a light microscopy and scanning electron
microscopy (SEM). Molecular characteristics, by confirmed their identification based on 16S rRNA gene sequences. The
microflora from both sampling locations was represented by three cyanobacteria morphospecies, Synechococcus sp.
(18.78%), Gloeocapsa sp. (12.20%), Chroococcus sp. (4.94%). The photomicrographs of all the dominant microalgae
recorded. Synechococcus sp. a cylindrical oblong elliptical unicellular or sometimes 2 to 4 cells as a result of cell
division. Under SEM Synechococcus sp. cells were cylindrical to oblong 2.6 μm in diameter and 3.8μm long. The cells
were enclosed in hyaline mucilage and with cells usually aggregating in large numbers to form colonies. Gloeocapsa
sp. cells were spherical, blue- green homogeneous contents, enclosed by wide colorless sheaths, of diameter 2-4μm, with
sheaths 4-8μm. Chloroococcus sp. cells were spherical, sometimes hemispherical after division, united cell in colonies
of two sheath, sheath is colorless and the individual cells were large; 20-28μm in diameter. According to DNA sequence
analysis, cyanobacteria l was closely related to Synechococcus sp, (99% similarity), cyanobacteria ll was found to be
similar to Gloeocapsa sp. (97% similarity) and cyanobacteria lll was Chroococcus sp. (95% similarity). This study,
used chemicals agents to control cyanobacteria from both sampling locations, by (NaOCl), (Ca(OCl)2) and (H₂O₂). The
result showed that, 5% H₂O₂, Ca(OCl)2 and NaOCl gave a significant difference (p˂0.05) on the growth of
cyanobacteria. However, the effect of 15% H₂O₂ was not significantly different (p˃0.05) with Ca(OCl)2 and NaOCl at
10, 15, and 30 minutes exposure, only 5 minutes exposure was significant (p˂0.05)
References
Price, L. (2006). Caves and caving in Malaysia. Caves and limestone hills of
malaysiahttp://www.cavesofmalaysia.com/
Barker, G., Piper, P. J., & Rabett, R. J. (2009). Zooarchaeology at the Niah Caves, Sarawak: Context and Research
Issues. International Journal of Osteoarchaeology, 19, 447–463.
Rajendran, R., & Prasad, N. (2012). A study on the indoor and outdoor microflora associated with the
biodeterioration of mural paintings at Sekharaipuram Vishnu temple, Adakkaputhur, Palakkad, Kerala, India
International Journal of Environmental Sciences and Research,1(4), 104-108 .
Sigee, D. C., Glenn, R., Andrews, M. J., E.G.Bellinger, R.D.Butler, H.A.S.Epton, et al. (1999). Biological control
of cyanobacteria: principles and possibilities. Hydrobiologia 161–172.
Muthukumar, C., Muralitharan, G., Vijayakumar, R., Panneerselvam, A., & Thajuddin, N. (2007). cyanobacterial
biodiversity from different freshwater ponds of thanjavur, tamilnadu (india). Acta botanica malacitana 17, 17-25.
Prescott, G. W. (1962). Algae of the western great lakes area with an lllustrated key to the genera of desmids and
freshwater diatoms U. S. A.: WM. C. Brown Company Publishers pp 932-977.
Fay, P., & Baalen, C. V. (1987). The cyanobacteria New York, Oxford: Elsevier 87-608.
Sincy, J. (2005). ecological and biochemical studies on cyanobacteria of cochin estuary and their application as
source of antioxidants. india. pp 159-218.
Garbacki, N., Ector, L., Kostikov, I., & Hoffmann, L. (1999). Contribution a` l‟e´tude de la flore des grottes de
Belgique Belgian Journal of Botany, 132(1), 43-76.
Smith, T., & Olson, R. (2007). A Taxonomic Survey of Lamp Flora (Algae and Cyanobacteria) in Electrically Lit
Passages within Mammoth Cave National Park, Kentucky. International Journal of Speleology, 36(2), 105-114.
Mulec, J., Kosi, G., & Vrhovsek, D. (2008). characterization of cave aerophytic algal communities and effects of
irradiance levels on production of pigments. Journal of cave and karst studies, 70 (1), 3–12.
Cahn, A. (Ed.). (1994). Proceedings of the 3rd World Conference on Detergents on Detetgents:Global
perspectives. World Conference on Detergents: Global Perspectives (3rd: 1993: Montreux, Switzerland, 279.
Al-Fawwaz, A. T. S. (2010). Biosorption of heavy metals from aqueous solutions by green algae isolated from
rivers in Penang, Malaysia. PhD Thesis, Universiti Sains Malaysia, Penang, Malaysia. 183pp.
Eliopoulos, G. M., & Eliopoulos, C. T. (1988). Antibiotic combinations: should they be tested. Clinical
microbiology 1(2)139–156.
Michael E. Klepser, Erika J. Ernst, Russell E. Lewis, Michael E. Ernst, & Pfaller, M. A. (1998). Influence of test
conditions on antifungal time-kill curve results: proposal for standardized methods. Antimicrob agents chemother. ,
(5), 1207–1212.
Iliopoulou-Georgoudaki, J., Pantazidou, A., & Theoulakis, P. (1993). an assessment of cleaning photoautotropic
micro lora: the case of "perama" cave, ioannina greece. Memoires e biospeologie, tome xx, 117-120.
Grobbelar, J. U. (2000). Lithophytic algae: A major threat to the karst formation of show caves. Journal of Applied
Phycology 12, 309–315.
Smith, G. M. (1950). The fresh water algae of the United States. New York Toronto London McGraw-Hill book
company. pp 537-725.
Prescott, G. W. (1962). Algae of the western great lakes area with an lllustrated key to the genera of desmids and
freshwater diatoms U. S. A.: WM. C. Brown Company Publishers pp 932-977.
Albertano, C. (1999). Structural interactions among epilithic cyanobacteria and heterotrophic microorganisms in
Roman hypogea Microbial Ecology, 38, 244-252.
Cennamo, P., Marzano, C., Ciniglia, C., Pinto, G., Cappelletti, P., Caputo, P., et al. (2011). A survey of the algal
flora of anthropogenic caves of campi flegrei (naples, italy) archeological district. Journal of cave and karst
studies, 74 (3), 243–250.
Lamprinou, V., Danielidis, D. B., Economou-Amilli, A., & Pantazidou, A. (2012). Distribution survey of
Cyanobacteria in three Greek caves of Peloponnese. International Journal of Speleology, 41(2), 267-272.
Hoffmann, L. (1989). Algae of terrestrial habitats. Botanical Review, 55, 77–105.
Scheerer, S., Ortega-Morales, O., Gaylarde, C., & Ahmad, M. (2009). Microbial deterioration of stone monuments-
-an updated overview. Advances in Applied Microbiology, 66, 97-139.
Marti´Nez, A., & Asencio, A. D. (2010). Distribution of cyanobacteria at the gelada cave(spain) by physical
parameters. Journal of cave and karst studies, 72(1), 11–20.
Czerwik-Marcinkowska, J., & Mrozińska, T. (2011). Algae and cyanobacteria in caves of the polish jura.
Polish Botanical Journal, 56(2), 203–243.
Cennamo, P., Marzano, C., Ciniglia, C., Pinto, G., Cappelletti, P., Caputo, P., et al. (2012). A survey of the algal
flora of anthropogenic caves of campi flegrei (naples, italy) archeological district.Journal of Cave and Karst
Studies 74(3), 243–250.
Rajendran, R., & Prasad, N. (2012). A study on the indoor and outdoor microflora associated with the
biodeterioration of mural paintings at Sekharaipuram Vishnu temple, Adakkaputhur, Palakkad, Kerala, India
International Journal of Environmental Sciences and Research,1(4), 104-108 .
Mulec, J. A., & Kosi, G. (2009). lampenflora algae and methods of growth control. Journal of cave and karst
studies, 71 (2), 109–115.
Cigna, A. A. (2011). Show cave development with special references to active caves. Sociedade Brasileira de
Espeleologia, 4(1) 16.
Faimon, J., Sctelcl, J., Kubesova´, S., & Zimak, J. (2003). Environmentally acceptable effect of hydrogen peroxide
on cave “lamp-flora”, calcite speleothems and limestones. Environmental Pollution, 122(3), 417- 422.
Goldstein JI, Newbury DE, Echlin P, Joy DC, Fiori C, Lifshin E. (1981). Scanning Electron Microscopy and XRay Microanalysis. Plenum Press New York and London.
Hayat MA. (1989). Principles and techniques of electron microscopy.Biological applications. Macmillan Press
LTD Houndmills and London..
Hernández Mariné M, Clavero E, Roldán M. (2004). Microscopy methods applied to research on cyanobacteria.
Asociación Ibérica de Limnología, Madrid Spain. (0213-8409).
Neilan BA, Jacobs D, Del Dot T, Blackall LL, Hawkins PR, Cox PT, et al. (1997). rRNA sequences and
evolutionary relationships among toxic and nontoxic cyanobacteria of the genus Microcystis. International Journal
of Systematic Bacteriology;47(693-697).
Nubel U, Garcia-Pichel F, Muyzer G. (1997). pcr primers to amplify 16s rrna genes from cyanobacteria. applied
and environmental microbiology. 63(8):3327-32.
Bosshard PP, Abels S, Zbinden R, Bo¨ttger EC, Altwegg M. (2003). Ribosomal DNA Sequencing for
Identification of Aerobic Gram-Positive Rods in the Clinical Laboratory (an 18-Month Evaluation). Journal Of
Clinical Microbiology. 41(9):4134-40.
Hebelka J. (2014). Methodology of Lampenflora removal in cave acceible for tourit. The cave adminitration of the
Czech Republic. Pruhonice:15.
Bertolani M, Cigna A, Maccio S, Morbidelli L, Sighinolfi G. (1991). The karst system “Grotta grande del vento –
Grotta del Fiume” and the conservation of its environment. In: Sauro U. (Ed.). In: Proceedings of the International
Conference on Environmental changes in karst areas. Dipartimento di Geografia UoP, editor.; p. 289-98
