Aims: Mangroves of Tanjung Lumpur, Kuantan, Pahang is considered as a relatively underexplored resource of actinomycetes. Based on the above perspective, a study was conducted on mangrove sediments of Tanjung Lumpur, Kuantan to isolate potential actinomycetes using several pretreatments and various selective media. Methodology and results: Sediments from five different sites at Tanjung Lumpur mangrove were collected and selectively pre-treated. The pretreated sediments were diluted and plated onto eight different selective media. A total of 172 potential actinomycetes were isolated from all the media. Antimicrobial activities of 61 selected strains were checked against 8 test microorganisms using cross streak method. Pretreatment of wet heat with seawater was the most effective method for the isolation of actinomycetes as it yielded a maximum of 105 actinomycete isolates and IM7 was the most suitable medium for actinomycete isolation with highest percentage of recovery (31 %). Forty three isolates (70.5 %) showed antimicrobial activities against one or more test microorganisms. Isolates IIUM B21 and IIUM B31 showed antimicrobial activity against all test microorganisms. Seven isolates showed antifungal activity as they inhibited only C. albicans. Ten isolates were randomly selected for identification based on partial sequences of 16S rRNA gene. Six isolates were found belong to the genus Streptomyces, two isolates belong to the genus Micromonospora and two isolates were identified as Rhodococcus spp. Conclusion, significance and impact of study: These findings revealed the potential of mangrove sediment of Tanjung Lumpur as an important source of actinomycetes with biosynthetic capabilities which might be beneficial to pharmaceutical industries.
Rhizophoraceae ; Actinobacteria

Nine secondary metabolites(S)-5-hydroxy-4-methylchroman-2-one(1), 4-methoxynaphthalene-1,5-diol(2), 8-methoxynaphthalene-1,7-diol(3), 1,8-dimethoxynaphthalene(4),(2R,4S)-2,3-dihydro-2-methyl-benzopyran-4,5-diol(5),(2R,4R)-3,4-dihydro-4-methoxy-2-methyl-2H-1-benzopyran-5-ol(6), 7-O-α-D-ribosyl-2,3-dihydro-5-hydroxy-2-methyl-chromen-4-one(7),(R)-3-methoxyl-1-(2,6-dihydroxyphenyl)-butan-1-one(8) and helicascolide A(9) were isolated from endophytic fungus Cladosporium sp. JJM22 by using column chromatographies of silica gel and ODS, and semi-preparative HPLC. Their structures were analyzed on the basis of spectroscopic and chemical data, especially NMR and MS. All isolated compounds were evaluated for their anti-inflammatory activities by examining the inhibitory activities on nitric oxide(NO) production induced by lipopolysaccharide in mouse macrophage RAW264.7 cells in vitro. Compounds 2-4 showed inhibitory activities.
Animals ; Benzopyrans ; Cladosporium ; Fungi ; Mice ; Molecular Structure ; Rhizophoraceae

Phytochemical investigation of Kandelia candel resulted in the isolation of six triterpenes (1 - 5) and two glyceryl glycosides (6 and 7) and their structures were determined by comparing the spectroscopic data with those of reported values. In present study, we described the inhibitory effects of fractions and isolated compounds from K. candel on pro-inflammatory cytokines (IL-12 p40, IL-6, and TNF-alpha) production in lipopolysaccharide (LPS) stimulated bone marrow-derived dendritic cells (BMDCs). Results indicated that compounds 3, 6, and 7 showed potent inhibition on IL-6 production (IC50 values at less than 0.5 microM, respectively). Meanwhile, compounds 6 and 7 exhibited strong inhibitory effects on the production of TNF-alpha (IC50 values of 1.7 +/- 0.1 and 5.5 +/- 0.2 microM). Compounds 1 and 3 were also showed the inhibitory effects on IL-12 p40 production (IC50 values of 8.9 +/- 0.4 and 3.3 +/- 0.1 microM, respectively).
Cytokines ; Dendritic Cells ; Glycosides* ; Interleukin-12 ; Interleukin-6 ; Rhizophoraceae* ; Triterpenes* ; Tumor Necrosis Factor-alpha

AIMTo investigate the secondary metabolites of the mangrove plant Ceriops tagal.

METHODSColumn chromatography techniques including HPLC were used for the separation and purification, and extensive spectral analysis including various 2D NMR spectra were employed for structure elucidation.

RESULTSNine compounds, namely, tagalsins A (1), ent-5alpha-dolabr-4 (18) -ene-15S,16-diol (2), squalene (3), betulinic acid (4), lup-20 (29) -en-3-on-28-oic acid (5), betulin (6), lup-20 (29) -en-3-on-28-ol (7), beta-sitosterol (8), n-hexacosanylferulate (9) were obtained. Of which 1 and 2 belong to dolabrane diterpene.

CONCLUSIONCompound 1 is a new compound, and 2 to 9 are isolated from this species for the first time.

Diterpenes ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Rhizophoraceae ; chemistry ; Squalene ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo investigate the chemical constituents in Bruguiera sexangula var. rhynchopetala.

METHODSilica gel flash chromatography together with Sephadex LH - 20 were performed for the isolation and purification of the petrol ether fraction of this plant, and the structures were elucidated by spectral analysis as well as the comparison of the spectral data with those reported in the literatures.

RESULTNine compounds were obtained and identified as lupeol (1), lupeone (2), trans-hydroxy-cinnamoyl ester of lupeol (3), taraxerone (4), beta-amyril-palmitate (5), squalene (6),beta-sitosterol (7), daucosterol (8) and 7alpha-hydroxy-sitosterol (9).

CONCLUSIONAll the compounds were first isolated from B. sexangula var. rhynchopetala.

Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Pentacyclic Triterpenes ; Plants, Medicinal ; chemistry ; Rhizophoraceae ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

Six hundred and eight fungi strains were isolated from seventy-eight samples of mangrove plants and soil that collected from Qinglan harbor. Cyctotoxic activity was detected by observing the growth inhibition or killing of the tumor cells under microscope. The result showed that 81 strains (about 13.32% of the total strains isolated) displayed cytotoxic activity against B16 tumor cell. The most fungi strains were isolated from mangrove plant Sonneratia alba, and most of cytotoxic active fungi strains were isolated from mangrove plant Heritiera littoralis.
Animals ; China ; Cytotoxicity Tests, Immunologic ; Fungi ; isolation & purification ; physiology ; Melanoma, Experimental ; pathology ; Mice ; Plant Roots ; microbiology ; Rhizophoraceae ; microbiology ; Soil Microbiology ; Tumor Cells, Cultured

In the search for novel potent fungi-derived bioactive compounds for bioinsecticide applications, crude ethyl acetate culture filtrate extracts from 110 mangrove fungal endophytes were screened for their toxicity. Toxicity tests of all extracts against brine shrimp (Artemia salina) larvae were performed. The extracts with the highest toxicity were further examined for insecticidal activity against Spodoptera litura larvae and acetylcholinesterase (AChE) inhibition activity. The results showed that the extracts of five isolates exhibited the highest toxicity to brine shrimp at 50% lethal concentration (LC50) values of 7.45 to 10.24 ppm. These five fungal isolates that obtained from Rhizophora mucronata were identified based on sequence data analysis of the internal transcribed spacer region of rDNA as Aspergillus oryzae (strain BPPTCC 6036), Emericella nidulans (strains BPPTCC 6035 and BPPTCC 6038), A. tamarii (strain BPPTCC 6037), and A. versicolor (strain BPPTCC 6039). The mean percentage of S. litura larval mortality following topical application of the five extracts ranged from 16.7% to 43.3%. In the AChE inhibition assay, the inhibition rates of the five extracts ranged from 40.7% to 48.9%, while eserine (positive control) had an inhibition rate of 96.8%, at a concentration of 100 ppm. The extracts used were crude extracts, so their potential as sources of AChE inhibition compounds makes them likely candidates as neurotoxins. The high-performance liquid chromatography profiles of the five extracts differed, indicating variations in their chemical constituents. This study highlights the potential of culture filtrate ethyl acetate extracts of mangrove fungal endophytes as a source of new potential bioactive compounds for bioinsecticide applications.
Acetylcholinesterase ; Artemia ; Aspergillus oryzae ; Chromatography, Liquid ; Complex Mixtures ; DNA, Ribosomal ; Emericella ; Endophytes* ; Larva ; Mortality ; Neurotoxins ; Physostigmine ; Rhizophoraceae ; Spodoptera ; Statistics as Topic ; Toxicity Tests

Soil salinity is an important issue, as most crop plants are low in salt tolerance. Salt tolerance, a complex, multifactorial, and multigenic process, has been known to be a quantitative trait. The identification of the salt stress responsive genes or salt tolerance genes is essential for the breeding programs. Most recent efforts have been focused on the products of structural genes (transport proteins, ion channels, enzymes of solute synthesis) while little attention were paid to the regulatory aspects of these proteins. Since the first aquaporin gene from plants was cloned and functionally expressed in 1993, there has been a growing interest in the molecular biology of MIPs (membrane intrinsic proteins) and their bearing on the biophysics of water flow across plant membranes. In the last decades, studies on Mangroves, a special kind of wood plants, grow in high-salt and flooding conditions have been concentrated almost exclusively on their physiological and ecological characteristics. Kandelia candel, one of the dominant species of mangroves along the Chinese coast, lacks salt glands or salt hairs used for removal of excess salt in other mangroves. This makes K. candel a perfect model to study the molecular mechanism of salt tolerance in mangrove plants. Using cDNA RDA, a cDNA-specific modification of genomic representational difference analysis, a series of salt responsive genes of Kandelia candel were cloned. Among these gene fragments, a 183 bp fragment (termed as SRGKC1) encoding a tonoplast intrinsic protein (TIP) in Kandelia candel (KCTIP1) was identified. Based on the sequence of SRGKC1, two gene specific primers were designed, and the 3' and 5' end of the KCTIP1 gene were obtained using the SMART RACE cDNA Amplification Kit. RACE products were purified from low-melting agarose, and sequenced directly with GSPs as the sequencing primers. A 500-bp fragment corresponding to the 3'end of this gene was obtained using the GSP1 primer, and a 690 bp fragment corresponding to the 5' end of this gene was obtained using the GSP2 primer. Two primers that flank the putative open reading frame (ORF) were designed to obtain the cDNA containing the complete ORF by RACE PCR reaction. The full-length cDNA of KCTIP1, containing a 756 bp open reading frame (ORF), was approximately 1.1 kb; the start codon was located at the nucleotides of 99-101 and stop codon at the nucleotides of 855-857 followed by a poly (A) tail. The KCTIP1 cDNA sequence in this research was released in GenBank with accession number AF521135. Using ExPASy Proteomics tools provided by EMBL, the isoelectric point and MWt of KCTIP1 are estimated as 5.77 and 26.3 kD respectively. Transmembrane prediction analysis revealed the deduced KCTIP1 protein sequence contains six transmembrane regions at amino acid residues of 20 - 42, 57 - 79, 86 - 108, 113 - 135, 142 - 164 and 217 - 239. Two highly conserved asparagine-proline-alanine (NPA) motifs were located at 85 - 87 and 199 - 201 amino acid residues respectively. KCTIP1 is also predicted to contain the Cys residue (Cys 118) that are shown to confer Hg-sensitivity in Arabidopsis gamma-TIP and delta-TIP. Similarity analysis showed that KCTIP1 shared 77% - 79% amino acid sequence identity with the TIPs from Vitis berlandieri, Brassica oleracea and Arabidopsis thaliana. Expression analyses indicated that KCTIP1 had different expression among species of Mangroves. Expressions of KCTIP1 in Kandelia candel, Rhizophora apoculata and Ceriops tagal were suppressed by salt, and were insensitive to salt stress in unknown species of Mangroves. Previous studied showed that salt conditions might result in large and rapid changes in extracellular water potential and serious disturbance to the cytoplasm. In order to compensate for this imbalance, the relative contribution of water channels to flow across the root could thus vary. K. candel is a species that is native to intertial zone of tropical and subtropical coast and is well-adapted to salt conditions. The coordinated down-regulation of aquaporins in this plant may decrease membrane water permeability and thus increase the cellular water conserva- tion during periods of salt stress. The results reported here are consistent with the postulated roles for tonoplast water channels in regulating the hydraulic permeability of the vacuolar membranes and in adjusting the water homeostasis of the protoplasm under various physiological conditions. The identification of KCTIP1 as one of salt-responsive genes implies that intracellular osmotic equilibration is a part of salt-tolerant mechanisms in Mangroves.
Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; DNA, Complementary ; genetics ; Membrane Proteins ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; Plant Proteins ; chemistry ; genetics ; Rhizophoraceae ; genetics ; Sequence Alignment

OBJECTIVETo identify the hepatoprotective and in vitro antioxidant activity of Lumnitzera racemosa (L. racemosa) leaf extract.

METHODSAnimals in Group 1 served as vehicle control, Group 2 served as hepatotoxin (CCL4 treated) group, Group 3 served as positive control (Silymarin) group, and Group 4, 5 and 6 served as (75, 150 and 300 mg/kg bw p.o.) L. racemosa leaf extract treated groups. Moreover, in vitro antioxidant DPPH, hydroxyl radical scavenging activity (HRSA), NO, ferric reducing antioxidant power (FRAP), lipid hydroperoxide (LPO) and super oxide dismutase (SOD) were also analyzed for the leaf extract.

RESULTSThe levels of the serum parameters such as serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP), bilirubin, cholesterol (CHL), sugar and lactate dehydrogenase (LDH) were significantly increased in CCL4 treated rats when compared with the control group (P<0.05). But the L. racemosa leaf extract treated rats showed maximum reduction of SGOT [(210.36±19.63) IU/L], SGPT [(82.37±13.87) IU/L], ALP [(197.63±23.43) IU/L], bilurubin [(2.15±0.84) mg/dL], cholesterol [(163.83±15.63) mg/dL], sugar [(93.00±7.65) mg/dL] and LDH [(1134.00±285.00) IU/L] were observed with the high dose (300 mg/kg bw) of leaf extract treated rats. Histopathological scores showed that, no visible changes were observed with high dose (300 mg/kg bw) of leaf extract treated rats except few mild necrosis. The IC50 values were observed as (56.37±4.87) µg/mL, (57.68±1.98) µg/mL, (64.15±2.90) µg/mL, (61.94±3.98) µg/mL, (94.53±1.68) µg/mL and (69.7±2.65) µg/mL for DPPH, HRSA, NO, FRAP, LPO and SOD radical scavenging activities, respectively.

CONCLUSIONSIn conclusion, the hepatoprotective effect of the L. racemosa leaf extract might be due to the presence of phenolic groups, terpenoids and alkaloids and in vitro antioxidant properties.

Animals ; Antioxidants ; pharmacology ; therapeutic use ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Liver ; chemistry ; drug effects ; pathology ; Male ; Plant Extracts ; chemistry ; Plant Leaves ; chemistry ; Protective Agents ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar ; Rhizophoraceae ; chemistry

OBJECTIVEThe aim of the present study was to isolate the anti-MRSA (Methicillin Resistant Staphylococcus aureus) molecule from the Mangrove symbiont Streptomyces and its biomedical studies in Zebrafish embryos.

METHODSMRSA was isolated from the pus samples of Colachal hospitals and confirmed by amplification of mecA gene. Anti-MRSA molecule producing strain was identified by 16s rRNA gene sequencing. Anti-MRSA compound production was optimized by Solid State Fermentation (SSF) and the purification of the active molecule was carried out by TLC and RP-HPLC. The inhibitory concentration and LC50 were calculated using Statistical software SPSS. The Biomedical studies including the cardiac assay and organ toxicity assessment were carried out in Zebrafish.

RESULTSThe bioactive anti-MRSA small molecule A2 was purified by TLC with Rf value of 0.37 with 1.389 retention time at RP-HPLC. The Inhibitory Concentration of the purified molecule A2 was 30 µg/mL but, the inhibitory concentration of the MRSA in the infected embryo was 32-34 µg/mL for TLC purified molecule A2 with LC50 mean value was 61.504 µg/mL. Zebrafish toxicity was assessed in 48-60 µg/mL by observing the physiological deformities and the heart beat rates (HBR) of embryos for anti MRSA molecule showed the mean of 41.33-41.67 HBR/15 seconds for 40 µg/mL and control was 42.33-42.67 for 15 seconds which significantly showed that the anti-MRSA molecule A2 did not affected the HBR.

CONCLUSIONSAnti-MRSA molecule from Streptomyces sp PVRK-1 was isolated and biomedical studies in Zebrafish model assessed that the molecule was non toxic at the minimal inhibitory concentration of MRSA.

Animals ; Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; toxicity ; Embryo, Nonmammalian ; drug effects ; pathology ; Heart Rate ; drug effects ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Microbial Sensitivity Tests ; Rhizophoraceae ; microbiology ; Streptomyces ; chemistry ; Zebrafish