Endophytic fungi have been explored not just for their ecological functions but also for their secondary metabolites as a new source of these pharmacologically active natural products. Accordingly, many structurally unique and biologically active compounds have been obtained from the cultures of endophytic fungi. Fusarium sp. and Lasiodiplodia theobromae were isolated from the root and stem of the mangrove plant Avicennia lanata, respectively, collected from Terengganu, Malaysia. High-resolution mass spectrometry and NMR spectroscopy were used as metabolomics profiling tools to identify and optimize the production of bioactive secondary metabolites in both strains at different growth stages and culture media. The spectral data was processed by utilizing Mzmine 2, a quantitative expression analysis software and an in house MS-Excel macro coupled with the Dictionary of Natural Products databases for dereplication studies. The investigation for the potential bioactive metabolites from a 15-day rice culture of Fusarium sp. yielded four 1,4- naphthoquinone with naphthazarin structures (1-4). On the other hand, the endophytic fungus L. theobromae grown on the 15-day solid rice culture produced dihydroisocoumarins (5-8). All the isolated compounds (1-8) showed significant activity against Trypanosoma brucei brucei with MIC values of 0.32-12.5 µM. Preliminary cytotoxicity screening against normal prostate cells (PNT2A) was also performed. All compounds exhibited low cytotoxicity, with compounds 3 and 4 showing the lowest cytotoxicity of only 22.3% and 38.6% of the control values at 100 µg/mL, respectively. Structure elucidation of the isolated secondary metabolites was achieved using 2D-NMR and HRESI-MS as well as comparison with literature data.
Chilling restrains the distribution of mangroves. We tested whether foliar phosphorus (P) fractions and gene expression are associated with cold tolerance in mangrove species. We exposed seedlings of six mangrove populations from different latitudes to favorable, chilling and recovery treatments, and measured their foliar P concentrations and fractions, photochemistry, nighttime respiration, and gene expression. A Kandelia obovata (KO; 26.45°N) population completely and a Bruguiera gymnorhiza (Guangxi) (BGG; 21.50°N) population partially (30%) survived chilling. Avicennia marina (24.29°N), and other B. gymnorhiza (26.66°N, 24.40°N, and 19.62°N) populations died after chilling. Photosystems of KO and photosystem I of BGG were least injured. During chilling, leaf P fractions, except nucleic acid P in three populations, declined and photoinhibition and nighttime respiration increased in all populations, with the greatest impact in B. gymnorhiza. Leaf nucleic acid P was positively correlated with photochemical efficiency during recovery and nighttime respiration across populations for each treatment. Relatively high concentrations of nucleic acid P and metabolite P were associated with stronger chilling tolerance in KO. Bruguiera gymnorhiza exhibited relatively low concentrations of organic P in favorable and chilling conditions, but its partially survived population showed stronger compensation in nucleic acid P and Pi concentrations and gene expression during recovery.
Wave energy and storm surges threaten coastal ecology and nearshore infrastructures. Although coastal structures are conventionally constructed to dampen the wave energy, they introduce tremendous damage to the ecology of the coast. To minimize environmental impact, ecofriendly coastal protection schemes should be introduced. In this paper, we discuss an example of an innovative mangrove rehabilitation attempt to restore the endangered mangroves on Carey Island, Malaysia. A submerged detached breakwater system was constructed to dampen the energy of wave and trap the sediments behind the structure. Further, a large number of mangrove seedlings were planted using different techniques. Further, we assess the possibility of success for a future mangrove rehabilitation project at the site in the context of sedimentology, bathymetry, and hydrogeochemistry. The assessment showed an increase in the amount of silt and clay, and the seabed was noticeably elevated. The nutrient concentration, the pH value, and the salinity index demonstrate that the site is conducive in establishing mangrove seedlings. As a result, we conclude that the site is now ready for attempts to rehabilitate the lost mangrove forest.
Bioaccumulation of lead and Copper in Avicennia marina and Rhizophora apiculata was studied. Samples of leaves, barks and roots were collected from Balok mangrove forest, Pahang. Pb and Cu accumulation was higher in Avicennia marina root tissue compared to bark and leaf but lower than surrounding sediment level. The average concentration of Pb in A. marina leaf, bark, root and sediment was observed to be 5.39 ppm, 3.63 ppm, 18.21 ppm and 23.13 ppm, and average Cu concentration was 4.13 ppm, 4.27 ppm, 4.81 ppm and 12.33 ppm, respectively. R. apiculata also showed higher concentration of Pb and Cu in root tissue compared to bark and leaf tissues but lower than surrounding sediment. The average concentration of Pb in R. apiculata leaf, bark, root and sediment was observed to be 4.30 ppm, 2.97 ppm, 22.45 ppm and 31.23 ppm, respectively. The average Cu concentration was 2.93 ppm, 4.71 ppm, 4.81 ppm and 15.52 ppm, respectively. Results of concentration factors (CF) showed that the accumulation of Pb and Cu was higher in A. marina than in R. apiculata.
A survey on coastal vegetation of Pahang in some localities from Sg. Balok in Kuantan southwards to Kuala Endau in Rompin was conducted from middle 2004 to early 2005. A total of 88 species of plants were recorded and identified mostly to the generic or species level. The usual vegetation types were sand and mudflat beach vegetations like Pandanus odoratissimus, Ipomoea pes capre and Casuarina equisetifolia and mangrove vegetation like Sonneratia caseolaris, Rhizophora mucronata and Avicennia marina. This paper also reports some unique heath-type forest vegetation like Hopea spp. in Menchali area.
Marine fungi on the whole remain understudied, especially in the highly diverse Southeast Asian region. We investigated the fungal communities associated with the mangrove tree Avicennia alba throughout Singapore and Peninsular Malaysia. At each sampling location, we examined ten individual trees, collecting leaves, fruits, pneumatophores, and an adjacent sediment sample from each plant. Amplicon sequencing of the fungal internal transcribed spacer 1 and subsequent analyses reveal significant differences in fungal communities collected from different locations and host structures. Mantel tests and multiple regression on distance matrices show a significant pattern of distance decay with samples collected close to one another having more similar fungal communities than those farther away. Submergence appears to drive part of the variation as host structures that are never submerged (leaves and fruits) have more similar fungal communities relative to those that are covered by water during high tide (pneumatophores and sediment). We suggest that fungi of terrestrial origins dominate structures that are not inundated by tidal regimes, while marine fungi dominate mangrove parts and sediments that are submerged by the incoming tide. Given the critical functions fungi play in all plants, and the important role they can have in determining the success of restoration schemes, we advocate that fungal community composition should be a key consideration in any mangrove restoration or rehabilitation project.
Horseshoe crabs are said to be declining worldwide. However, there is still no published report on the status of horseshoe crabs in Malaysia. Thus, we report here eight informative microsatellite markers that were developed using the 5'-anchored ISSR-PCR enrichment procedure to diagnose the population genetic structure of the mangrove horseshoe crab, Carcinoscorpius rotundicauda from Peninsular Malaysia. This set of markers was tested on 127 samples and showed polymorphism in this species. Hence they should be useful in future essential population genetic studies of these living fossils in the Southeast Asian region.
A study on the importance of Xylocopa varipuncta (Hymenoptera: Apidae) as pollination agent was conducted at the Setiu Mangrove Forest, Terengganu from September to December 2010. The objective of this study was to identify the pollens collected by carpenter bees (X. varipuncta) in the mangrove community of Setiu Wetlands. A total of 35 types of pollens were collected from the body of X. varipuncta and only 10 types of the pollens were successfully identified. The identified pollens were of Avicennia alba, Lumnitzera racemosa, Sonneratia caseolaris, S. ovata and Rhizophora apiculata from exclusive mangroves, while Suregada multiflora, Melaleuca cajuputi, Derris trifoliata, Acacia auriculiformis and Hibiscus tiliaceus were from non-exclusive mangroves. Melaleuca cajuputi was the highest number of pollen carried by X. varipuncta. This study showed that X. varipuncta is an important pollen carrier in the mangrove community of Setiu Wetlands, Terengganu.
Mangrove plants are endowed with various biologically active compounds which have potent antibacterial and antioxidant properties. In present study, a bioactivity-guided fractionation for antibacterial and antioxidant active metabolites from the twigs of Avicennia officinalis collected from Kuala Selangor Nature Park, Selangor, Malaysia gave 13 major fractions. The antibacterial activity of A. officinalis fractions using well-diffusion showed strong selectivity on the Gram-positive bacteria (Staphylococcus epidermidis, S. aureus and Bacillus subtilis) with minimum inhibition concentration (MIC) values of 0.156-5.00 mg/mL. However, no antibacterial activities were observed on the Gram-negative bacteria (Vibrio cholera, Enterobacter cloacae and Escherichia coli). The active antibacterial fractions were further isolated using several chromatographic techniques to give two naphthofuranquinones, namely, avicenol C (1) and stenocarpoquinone B (2). Meanwhile, the antioxidant activity of A. officinalis fractions were evaluated using DPPH radical scavenging assay exhibited low antioxidant activities. Molecular structure of the naphthofuranquinones was elucidated using 1 D and 2 D NMR spectroscopy.
The discovery of new secondary metabolites from natural origins has become more challenging in natural products research. Different approaches have been applied to target the isolation of new bioactive metabolites from plant extracts. In this study, bioactive natural products were isolated from the crude organic extract of the mangrove plant Avicennia lanata collected from the east coast of Peninsular Malaysia in the Setiu Wetlands, Terengganu, using HRESI-LCMS-based metabolomics-guided isolation and fractionation. Isolation work on the crude extract A. lanata used high-throughput chromatographic techniques to give two new naphthofuranquinone derivatives, hydroxyavicenol C (1) and glycosemiquinone (2), along with the known compounds avicenol C (3), avicequinone C (4), glycoquinone (5), taraxerone (6), taraxerol (7), β-sitosterol (8) and stigmasterol (9). The elucidation and identification of the targeted bioactive compounds used 1D and 2D-NMR and mass spectrometry. Except for 6-9, all isolated naphthoquinone compounds (1-5) from the mangrove plant A. lanata showed significant anti-trypanosomal activity on Trypanosoma brucei brucei with MIC values of 3.12-12.5 μM. Preliminary cytotoxicity screening against normal prostate cells (PNT2A) was also performed. All compounds exhibited low cytotoxicity, with compounds 3 and 4 showing moderate cytotoxicity of 78.3% and 68.6% of the control values at 100 μg/mL, respectively.
Strain MUSC 117(T) was isolated from mangrove soil of the Tanjung Lumpur forest in Pahang, Malaysia. This bacterium was yellowish-white pigmented, Gram-staining-positive, rod-coccus shaped and non-motile. On the basis of 16S rRNA gene sequence, strain MUSC 117(T) exhibited highest sequence similarity to Sinomonas atrocyanea DSM 20127(T) (98.0 %), Sinomonas albida LC13(T) (97.9 %) and Sinomonas soli CW 59(T) (97.8 %), and lower (<97.6 %) sequence similarity to other species of the genus Sinomonas. DNA-DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 27 %) between strain MUSC 117(T) and closely related species. Chemotaxonomically, the peptidoglycan type was A3α, containing the amino acids lysine, serine, glycine, alanine, glutamic acid and muramic acid. The whole-cell sugars detected were rhamnose, ribose, glucose, galactose and a smaller amount of mannose. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and five unidentified glycolipids. The major fatty acids (>10.0 %) of the cell membrane were anteiso-C15 : 0 (39.4 %), C18 : 1ω7c (17.7 %), anteiso-C17 : 0 (17.2 %) and iso-C16 : 0 (11.4 %). The predominant respiratory quinones detected were MK-9(H2) and MK-9. The DNA G+C content was 67.3 mol%. A comparison of BOX-PCR fingerprints indicated that strain MUSC 117(T) represented a unique DNA profile. Results based on a polyphasic approach showed that strain MUSC 117(T) represents a novel species of the genus Sinomonas, for which the name Sinomonas humi sp. nov. is proposed. The type strain of Sinomonas humi sp. nov. is MUSC 117(T) ( = DSM 29362(T) = MCCC 1K00410(T) = NBRC 110653(T)).
Strain MUSC 115(T) was isolated from mangrove soil of the Tanjung Lumpur river in the state of Pahang, Peninsular Malaysia. Cells of this strain stained Gram-positive and were non-spore-forming, short rods that formed yellowish-white colonies on different agar media. The taxonomy of strain MUSC 115(T) was studied by a polyphasic approach, and the organism showed a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Microbacterium. The cell-wall peptidoglycan was of type B2β, containing the amino acids ornithine, alanine, glycine, glutamic acid and homoserine. The muramic acid was of the N-glycolyl form. The predominant menaquinones detected were MK-12, MK-13 and MK-11. The polar lipids consisted of phosphatidylglycerol, phosphoglycolipid, diphosphatidylglycerol, two unidentified lipids, three unidentified phospholipids and four unidentified glycolipids. The major fatty acids of the cell membrane were anteiso-C15:0 and anteiso-C17:0. The whole-cell sugars detected were ribose, glucose, mannose and galactose. Based on the 16S rRNA gene sequence, strain MUSC 115(T) showed the highest sequence similarity to Microbacterium immunditiarum SK 18(T) (98.1%), M. ulmi XIL02(T) (97.8%) and M. arborescens DSM 20754(T) (97.5%) and lower sequence similarity to strains of other species of the genus Microbacterium. DNA-DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 24%) between strain MUSC 115(T) and the type strains of closely related species. Furthermore, BOX-PCR fingerprint comparison also indicated that strain MUSC 115(T) represented a unique DNA profile. The DNA G+C content determined was 70.9 ± 0.7 mol%, which is lower than that of M. immunditiarum SK 18(T). Based on the combination of genotypic and phenotypic data, it is proposed that strain MUSC 115(T) represents a novel species of the genus Microbacterium, for which the name Microbacterium mangrovi sp. nov. is proposed. The type strain is MUSC 115(T) ( = MCCC 1K00251(T) = DSM 28240(T) = NBRC 110089(T)).
The impact of floating net cages culturing the seabass, Lates calcarifer, on planktonic processes and water chemistry in two heavily used mangrove estuaries in Malaysia was examined. Concentrations of dissolved inorganic and particulate nutrients were usually greater in cage vs. adjacent (approximately 100 m) non-cage waters, although most variability in water-column chemistry related to water depth and tides. There were few consistent differences in plankton abundance, production or respiration between cage and non-cage sites. Rates of primary production were low compared with rates of pelagic mineralization reflecting high suspended loads coupled with large inputs of organic matter from mangrove forests, fishing villages, fish cages, pig farms and other industries within the catchment. Our preliminary sampling did not reveal any large-scale eutrophication due to the cages. A crude estimate of the contribution of fish cage inputs to the estuaries shows that fish cages contribute only approximately 2% of C but greater percentages of N (32-36%) and P (83-99%) to these waters relative to phytoplankton and mangrove inputs. Isolating and detecting impacts of cage culture in such heavily used waterways--a situation typical of most mangrove estuaries in Southeast Asia--are constrained by a background of large, highly variable fluxes of organic material derived from extensive mangrove forests and other human activities.
A novel, rod-shaped, Gram-stain-negative, halophilic and non-motile bacterium, designated CCB-MM1T, was isolated from a sample of estuarine sediment collected from Matang Mangrove Forest, Malaysia. The cells possessed a rod-coccus cell cycle in association with growth phase and formed aggregates. Strain CCB-MM1T was both catalase and oxidase positive, and able to degrade starch. Optimum growth occurred at 30 °C and pH 7.0 in the presence of 2-3 % (w/v) NaCl. The 16S rRNA gene sequence of strain CCB-MM1T showed 98.12, 97.46 and 97.33 % sequence similarity with Microbulbifer rhizosphaerae Cs16bT, Microbulbifer maritimus TF-17T and Microbulbifergwangyangensis GY2T respectively. Strain CCB-MM1T and M. rhizosphaerae Cs16bT formed a cluster in the phylogenetic tree. The major cellular fatty acids were iso-C17 : 1 ω9c and iso-C15 : 0, and the total polar lipid profile consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphoaminolipid, two unidentified lipids, an unidentified glycolipid and an unidentified aminolipid. The major respiratory quinone was ubiquinone Q-8 and the genomic DNA G+C content of the strain was 58.9 mol%. On the basis of the phylogenetic, phenotypic and genotypic data presented here, strain CCB-MM1T represents a novel species of the genus Microbulbifer, for which the name Microbulbiferaggregans sp. nov. is proposed. The type strain is CCB-MM1T (=LMG 29920T=JCM 31875T).
Two novel actinobacteria, strains MUSC 135(T) and MUSC 137, were isolated from mangrove soil at Tanjung Lumpur, Malaysia. The 16S rRNA gene sequence similarity and DNA-DNA relatedness between strains MUSC 135(T) and MUSC 137 were 100 % and 83±3.2 %, confirming that these two strains should be classified in the same species. Strain MUSC 135(T) exhibited a broad-spectrum bacteriocin against the pathogens meticillin-resistant Staphylococcus aureus (MRSA) strain ATCC BAA-44, Salmonella typhi ATCC 19430(T) and Aeromonas hydrophila ATCC 7966(T). A polyphasic approach was used to study the taxonomy of MUSC 135(T), and it showed a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Streptomyces. The diamino acid of the cell-wall peptidoglycan was ll-diaminopimelic acid. The predominant menaquinones were MK-9(H6), MK-9(H4) and MK-9(H8). Polar lipids detected were a lipid, an aminolipid, a phospholipid, phosphatidylinositol, phosphatidylethanolamine and two glycolipids. The predominant cellular fatty acids (>10.0 %) were anteiso-C15 : 0 (20.8 %), iso-C16 : 0 (18.0 %), iso-C15 : 0 (12.2 %) and anteiso-C17 : 0 (11.6 %). The whole-cell sugars were ribose, glucose and mannose. These results suggested that MUSC 135(T) should be placed within the genus Streptomyces. Phylogenetic analysis based on the 16S rRNA gene sequence exhibited that the most closely related strains were Streptomyces cinereospinus NBRC 15397(T) (99.18 % similarity), Streptomyces mexicanus NBRC 100915(T) (99.17 %) and Streptomyces coeruleofuscus NBRC 12757(T) (98.97 %). DNA-DNA relatedness between MUSC 135(T) and closely related type strains ranged from 26.3±2.1 to 49.6±2.5 %. BOX-PCR fingerprint comparisons showed that MUSC 135(T) exhibited a unique DNA profile. The DNA G+C content determined was 70.7±0.3 mol%. Based on our polyphasic study of MUSC 135(T), the strain merits assignment to a novel species, for which the name Streptomyces pluripotens sp. nov. is proposed. The type strain is MUSC 135(T) ( = MCCC 1K00252(T) = DSM 42140(T)).