Saxitoxins (STXs) constitute a family of potent sodium channel blocking toxins, causative agents of paralytic shellfish poisoning (PSP), and are produced by several species of marine dinoflagellates and cyanobacteria. Two STX-core genes, sxtA and sxtG, have been well elucidated in Alexandrium but the expression of these genes under various nutritional modes in tropical species remains unclear. This study investigates the physiological responses of a tropical Pacific strain of Alexandrium minutum growing with nitrate or ammonium, and with various nitrogen to phosphorus (N:P) supply ratios. The transcriptional responses of the sxt genes were observed. Likewise, a putative sxtI encoding O-carbamoyltransferase (herein designated as AmsxtI) was recovered from the transcriptomic data, and its expression was investigated. The results revealed that the cellular toxin quota (Qt) was higher in P-depleted, nitrate-grown cultures. With cultures at similar N:P (<16), cells grown with excess ammonium showed a higher Qt than those grown with nitrate. sxtA1 was not expressed under any culture conditions, suggesting that this gene might not be involved in STX biosynthesis by this strain. Conversely, sxtA4 and sxtG showed positive correlations with Qt, and were up-regulated in P-depleted, nitrate-grown cultures and with excess ambient ammonium. On the other hand, AmsxtI was expressed only when induced by P-depletion, suggesting that this gene may play an important role in P-recycling metabolism, while simultaneously enhancing toxin production.
Natural and environmental-friendly disposal of wastewater sludge is a great concern. Recently, biological treatment has played prominent roles in bioremediation of complex hydrocarbon- rich contaminants. Composting is quite an old biological-based process that is being practiced but it could not create a great impact in the minds of concerned researchers. The present study was conducted to evaluate the feasibility of the solid-state bioconversion (SSB) processes in the biodegradation of wastewater sludge by exploiting this promising technique to rejuvenate the conventional process. The Indah Water Konsortium (IWK) domestic wastewater treatment plant (DWTP) sludge was considered for evaluation of SSB by monitoring the microbial growth and its subsequent roles in biodegradation under two conditions: (i) flask (F) and (ii) composting bin (CB) cultures. Sterile and semi-sterile environments were allowed in the F and the CB, respectively, using two mixed fungal cultures, Trichoderma harzianum with Phanerochaete chrysosporium 2094 (T/P) and T. harzianum with Mucor hiemalis (T/M) and two bulking materials, sawdust (SD) and rice straw (RS). The significant growth and multiplication of both the mixed fungal cultures were reflected in soluble protein, glucosamine and color intensity measurement of the water extract. The color intensity and pH of the water extract significantly increased and supported the higher growth of microbes and bioconversion. The most encouraging results of microbial growth and subsequent bioconversion were exhibited in the RS than the SD. A comparatively higher decrease of organic matter (OM) % and C/N ratio were attained in the CB than the F, which implied a higher bioconversion. But the measurement of soluble protein, glucosamine and color intensity exhibited higher values in the F than the CB. The final pH drop was higher in the CB than the F, which implied that a higher nitrification occurred in the CB associated with a higher release of H+ ions. Both the mixed cultures performed almost equal roles in all cases except the changes in moisture content.
Many wastewater treatment plants (WWTPs) operating in biological nitrogen removal activated sludge process in the tropics are facing the pressure of increasingly stringent effluent standards while seeking solutions to reduce the plants' energy consumption and operating cost. This study investigated the feasibility of applying low-dissolved oxygen (low-DO) nitrification and utilizing slowly-biodegradable chemical oxygen demand (sbCOD) for denitrification, which helps to reduce energy usage and operating cost in treating low soluble COD-to-nitrogen tropical wastewater. The tropical wastewater was first characterized using wastewater fractionation and respirometry batch tests. Then, a lab-scale sequencing batch reactor (SBR) was operated to evaluate the long-term stability of low-DO nitrification and utilizing sbCOD for denitrification in an anoxic-oxic (AO) process treating tropical wastewater. The wastewater fractionation experiment revealed that particulate settleable solids (PSS) in the wastewater provided slowly-biodegradable COD (sbCOD), which made up the major part (51 ± 10%) of the total COD. The PSS hydrolysis rate constant at tropical temperature (30 °C) was 2.5 times higher than that at 20 °C, suggesting that sbCOD may be utilized for denitrification. During the SBR operation, high nitrification efficiency (93 ± 6%) was attained at low-DO condition (0.9 ± 0.1 mg O2/L). Utilizing sbCOD for post-anoxic denitrification in the SBR reduced the effluent nitrate concentration. Quantitative polymerase chain reaction, 16S rRNA amplicon sequencing and fluorescence in-situ hybridization revealed that the genus Nitrospira was a dominant nitrifier. 16S rRNA amplicon sequencing result suggested that 50% of the Nitrospira-related operational taxonomic units were affiliated with comammox, which may imply that the low-DO condition and the warm wastewater promoted their growth. The nitrogen removal in a tropical AO process was enhanced by incorporating low-DO nitrification and utilizing sbCOD for post-anoxic denitrification, which contributes to an improved energy sustainability of WWTPs.
Acetone-butanol-ethanol (ABE) production from renewable resources has been widely reported. In this study, Clostridium butyricum EB6 was employed for ABE fermentation using fermentable sugar derived from treated oil palm empty fruit bunch (OPEFB). A higher amount of ABE (2.61 g/l) was produced in a fermentation using treated OPEFB as the substrate when compared to a glucose based medium that produced 0.24 g/l at pH 5.5. ABE production was increased to 3.47 g/l with a yield of 0.24 g/g at pH 6.0. The fermentation using limited nitrogen concentration of 3 g/l improved the ABE yield by 64%. The study showed that OPEFB has the potential to be applied for renewable ABE production by C. butyricum EB6.
A split plot 3 by 4 experiment was designed to characterize the relationship between production of gluthatione (GSH), oxidized gluthatione (GSSG), total flavonoid, anthocyanin, ascorbic acid and antioxidant activities (FRAP and DPPH) in three varieties of Labisia pumila Blume, namely the varieties alata, pumila and lanceolata, under four levels of nitrogen fertilization (0, 90, 180 and 270 kg N/ha) for 15 weeks. The treatment effects were solely contributed by nitrogen application; there was neither varietal nor interaction effects observed. As the nitrogen levels decreased from 270 to 0 kg N/ha, the production of GSH and GSSG, anthocyanin, total flavonoid and ascorbic acid increased steadily. At the highest nitrogen treatment level, L. pumila exhibited significantly lower antioxidant activities (DPPH and FRAP) than those exposed to limited nitrogen growing conditions. Significant positive correlation was obtained between antioxidant activities (DPPH and FRAP), total flavonoid, GSH, GSSG, anthocyanin and ascorbic acid suggesting that an increase in the antioxidative activities in L. pumila under low nitrogen fertilization could be attributed to higher contents of these compounds. From this observation, it could be concluded that in order to avoid negative effects on the quality of L. pumila, it is advisable to avoid excessive application of nitrogen fertilizer when cultivating the herb for its medicinal use.
A split plot 3 by 3 experiment was designed to investigate the relationships among production of primary metabolites (soluble sugar and starch), secondary metabolites (total flavonoids, TF; total phenolics, TP), phenylalanine lyase (PAL) activity (EC 4.3.1.5), protein and antioxidant activity (FRAP) of three progenies of oil palm seedlings, namely Deli AVROS, Deli Yangambi and Deli URT, under three levels of CO₂ enrichment (400, 800 and 1,200 μmol·mol⁻¹) for 15 weeks of exposure. During the study, the treatment effects were solely contributed by CO₂ enrichment levels; no progenies and interaction effects were observed. As CO₂ levels increased from 400 to 1,200 μmol·mol⁻¹, the production of carbohydrate increased steadily, especially for starch more than soluble sugar. The production of total flavonoids and phenolics contents, were the highest under 1,200 and lowest at 400 μmol·mol⁻¹. It was found that PAL activity was peaked under 1,200 μmol·mol⁻¹ followed by 800 μmol·mol⁻¹ and 400 μmol·mol⁻¹. However, soluble protein was highest under 400 μmol·mol⁻¹ and lowest under 1,200 μmol·mol⁻¹. The sucrose/starch ratio, i.e., the indication of sucrose phosphate synthase actvity (EC 2.4.1.14) was found to be lowest as CO₂ concentration increased from 400 > 800 > 1,200 μmol·mol⁻¹. The antioxidant activity, as determined by the ferric reducing/antioxidant potential (FRAP) activity, increased with increasing CO₂ levels, and was significantly lower than vitamin C and α-tocopherol but higher than butylated hydroxytoluene (BHT). Correlation analysis revealed that nitrogen has a significant negative correlation with carbohydrate, secondary metabolites and FRAP activity indicating up-regulation of production of carbohydrate, secondary metabolites and antioxidant activity of oil palm seedling under elevated CO₂ was due to reduction in nitrogen content in oil palm seedling expose to high CO₂ levels.
Kacip Fatimah (Labisia pumila Blume), one of the most famous and widely used herbs, especially in Southeast Asia, is found to have interesting bioactive compounds and displays health promoting properties. In this study, the antioxidant activities of the methanol extracts of leaves, stems and roots of three varieties of L. pumila (var. alata, pumila and lanceolata) were evaluated in an effort to compare and validate the medicinal potential of this indigenous Malaysian herb species. The antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, as well as the total amount of phenolics and flavonoids were the highest in the leaves, followed by the stems and roots in all the varieties. A similar trend was displayed by the ferric reducing antioxidant potential (FRAP) activity, suggesting that the L. pumila varieties possess high foliar antioxidant properties. At low FRAP activity concentrations, the values of the leaves' inhibition activity in the three varieties were significantly higher than those of the stems and roots, with var. alata exhibiting higher antioxidant activities and total contents of phenolics and flavonoids compared to the varieties pumila and lanceolata. The high production of secondary metabolites and antioxidant activities in var. alata were firmly related to low nitrogen content and high C/N ratio in plant parts. The study also demonstrated a positive correlation between secondary metabolite content and antioxidant activities, and revealed that the consumption of L. pumila could exert several beneficial effects by virtue of its antioxidant activity.
A split plot 3 by 4 experiment was designed to examine the impact of 15-week variable levels of nitrogen fertilization (0, 90, 180 and 270 kg N/ha) on the characteristics of total flavonoids (TF), total phenolics (TP), total non structurable carbohydrate (TNC), net assimilation rate, leaf chlorophyll content, carbon to nitrogen ratio (C/N), phenyl alanine lyase activity (PAL) and protein content, and their relationships, in three varieties of Labisia pumila Blume (alata, pumila and lanceolata). The treatment effects were solely contributed by nitrogen application; there was neither varietal nor interaction effect observed. As nitrogen levels increased from 0 to 270 kg N/ha, the production of TNC was found to decrease steadily. Production of TF and TP reached their peaks under 0 followed by 90, 180 and 270 kg N/ha treatment. However, net assimilation rate was enhanced as nitrogen fertilization increased from 0 to 270 kg N/ha. The increase in production of TP and TF under low nitrogen levels (0 and 90 kg N/ha) was found to be correlated with enhanced PAL activity. The enhancement in PAL activity was followed by reduction in production of soluble protein under low nitrogen fertilization indicating more availability of amino acid phenyl alanine (phe) under low nitrogen content that stimulate the production of carbon based secondary metabolites (CBSM). The latter was manifested by high C/N ratio in L. pumila plants.
A factorial split plot 4 × 3 experiment was designed to examine and characterize the relationship among production of secondary metabolites (total phenolics, TP; total flavonoids, TF), carbohydrate content and photosynthesis of three varieties of the Malaysian medicinal herb Labisia pumila Benth. namely the varieties alata, pumila and lanceolata under CO(2) enrichment (1,200 µmol mol(-1)) combined with four levels of nitrogen fertilization (0, 90, 180 and 270 kg N ha(-1)). No varietal differences were observed, however, as the levels of nitrogen increased from 0 to 270 kg N ha(-1), the production of TP and TF decreased in the order leaves>roots>stems. The production of TP and TF was related to increased total non structural carbohydrate (TNC), where the increase in starch content was larger than that in sugar concentration. Nevertheless, the regression analysis exhibited a higher influence of soluble sugar concentration (r(2) = 0.88) than starch on TP and TF biosynthesis. Photosynthesis, on the other hand, displayed a significant negative relationship with TP and TF production (r(2) = -0.87). A decrease in photosynthetic rate with increasing secondary metabolites might be due to an increase in the shikimic acid pathway that results in enhanced production of TP and TF. Chlorophyll content exhibited very significant negative relationships with total soluble sugar, starch and total non structural carbohydrate.
A two-staged engineered wetland-based system was designed and constructed to treat raw domestic septage. Hydraulic loading rates (HLRs) of 8.75 and 17.5 cm/d were studied with four and eight daily dosings at the second stage of the system to investigate the influence of the regimes on septage treatment. Removal of organic matter (OM) was found to be HLR dependent, where the results indicated that the increase of HLR from 8.75 to 17.5 cm/d impaired the overall level of treatment in the wetland units. Effluent of wetland fed at HLR 17.5 cm/d presented significantly lower oxygen reduction potential and dissolved oxygen values than wetland fed at 8.75 cm/d, indicative of the occurrence of less aerobic and reductive conditions in the bed. The reoxygenation capability of the wetland units was found to be heavily affected by the dosing frequency especially under high hydraulic load (17.5 cm/d). NH3-N degradation was found to decrease with statistical importance when the wetland was flushed two times more frequently with smaller batches of influent. The number of hydraulic load fractionings did not seem to affect the level of treatments of OM and ammonia for both the wetlands fed under the lower HLR of 8.75 cm/d. Prediction of hydraulic limits and management of the feeding strategies are important in the vertical type of engineered wetlands to guarantee the treatment performance and minimize the chances of filter clogging.
Variations in leaf photosynthetic, morphological and biochemical properties with increasing plant height from seedlings to emergent trees were investigated in five dipterocarp species in a Malaysian tropical rain forest. Canopy openness increased significantly with tree height. Photosynthetic properties, such as photosynthetic capacity at light saturation, light compensation point, maximum rate of carboxylation and maximum rate of photosynthetic electron transport, all increased significantly with tree height. Leaf morphological and biochemical traits, such as leaf mass per area, palisade layer thickness, nitrogen concentration per unit area, chlorophyll concentration per unit dry mass and chlorophyll to nitrogen ratio, also changed significantly with tree height. Leaf properties had simple and significant relationships with tree height, with few intra- and interspecies differences. Our results therefore suggest that the photosynthetic capacity of dipterocarp trees depends on tree height, and that the trees adapt to the light environment by adjusting their leaf morphological and biochemical properties. These results should aid in developing models that can accurately estimate carbon dioxide flux and biomass production in tropical rain forests.
Earthworms are commonly referred as environmental engineers and their guts are often compared with chemical reactors. However, modeling experiments to substantiate it are lacking. The aim of this study was to use established reactor models, particularly PFR, on the gut of the vermicomposting earthworm Eudrilus eugeniae to understand more on its digestion. To achieve the objective, a mathematical model based on first-order kinetics was framed and used to determine the pattern of digestion rates of nutrient indicators, namely total carbon (%), total nitrogen (%), C/N ratio, 13C (‰), and 15N (‰) at five intersections (pre-intestine, foregut, midgut A, midgut B, and hindgut) along the gut of E. eugeniae. The experimental results revealed that the concentrations of TC, TN, 13C, and 15N decreased during gut transit, whereas C/N ratio increased. The first-order model demonstrated that all the nutrients exhibit a linear pattern of digestion during gut transit, which supports the PFR model. On this basis, the present study concludes that the gut of E. eugeniae functions as PFR.
The growth and lectin production of Ganoderma applanatum, a white rot fungus, was optimized in broth cultures. The fungus was found to have a higher growth rate and higher lectin activity when grown in a medium adjusted to pH 6.5 at 26°C under stationary conditions. Expression of lectin activity started in 5-day-old mycelial culture; maximum activity was expressed after the 15th day of incubation. Among the various carbon and nitrogen sources tested, the carbon source sucrose and the nitrogen source yeast extract support maximum growth and lectin production. Lectin from G. applanatum was purified by ammonium sulfate precipitation and ion exchange chromatography. The purified fraction revealed a single band with a molecular weight of 35.0 kDa. Moreover, carbohydrates such as mannitol, glucose, sucrose, maltose, mannose, galactose, sorbose, and fructose were found to inhibit the hemagglutinating activity of the lectin. The purified lectins from G. applanatum contain cytotoxic and proapoptotic activities against HT-29 colon adenocarcinoma cells.
Present study aims to optimize the production of starch and total carbohydrates from Arthrospira platensis. Growing concerns toward unprecedented environmental issues associated with plastic pollution has created a tremendous impetus to develop new biomaterials for the production of bioplastic. Starch-based biopolymers from algae serve as sustainable feedstock for thermoplastic starch production due to their abundant availability and low cost. A. platensis was cultivated in Zarrouk's medium at 32 ± 1°C and exposed to red light with a photoperiod of 12:12 hr light/dark. Growth kinetics studies showed that the maximum specific growth rate (μmax ) obtained was 0.059 day-1 with the doubling time (td ) of 11.748 days. Subsequently, Zarrouk's medium with different concentrations of sulfur, phosphorus and nitrogen was prepared to establish the nutrient-limiting conditions to enhance the accumulation of starch and total carbohydrates. In this study, the highest starch accumulated was 6.406 ± 0.622 mg L-1 under optimized phosphorus limitation (0.025 g L-1 ) conditions. Nitrogen limitation (0.250 g L-1 ) results demonstrated significant influenced (p
The effects of different carbon and nitrogen sources on phytase production by Mitsuokella jalaludinii were evaluated and the optimization of rice bran (RB) and soybean milk (SM) concentrations in the medium for phytase production was also determined.
The availability of paddy husk from rice processing plants remains high owing to increase in the worldwide rice consumption. Increasing demand for chicken products leads to poultry wastes production. Co-composting of the aforementioned wastes could solve the indiscriminate disposal of these wastes. Thus, co-composting of paddy husk and chicken slurry with clinoptilolite zeolite and urea as additive was carried out. Clinoptilolite zeolite was used to enhance ammonium and nitrate retention in the compost. Temperature of the compost was monitored three times daily for 55 days. Cation exchange capacity, organic matter, ash, humic acids, pH, total C, N, C/N ratio; total P, exchangeable Ca, Mg, K, NH4+, NO3-, and heavy metals contents were determined using standard procedures. pH, total N, humic acids, ash, NH4+, NO3-, P, Ca, Mg, and K contents increased but the salinity, heavy metals contents, and microbial population were low after the co-composting process. Zea mays L. (test crop) seed germination rate in distilled water and the compost were not significantly different. Growth of Spinach oleracea (test crop) on a peat-based growing medium and the compost was also not significantly different. These findings were possible because the clinoptilolite zeolite used in co-composting reduced accumulation of heavy metals that may have damage effects on the test crops. Mature compost with good agronomic properties can be produced by co-composting chicken slurry and paddy husk using clinoptilolite zeolite and urea as additives.
Although freshwater biomes cover less than 1% of the Earth's surface, they have disproportionate ecological significances. Attempts to study the taxonomy and function of freshwater microbiota are currently limited to samples collected from temperate lakes. In this study, we investigated samples from the photic and aphotic of an aquaculture site (disturbed) of Temengor Lake, a tropical lake in comparison with the undisturbed site of the lake using 16S rRNA amplicon and shotgun metagenomic approaches. Vertical changes in bacterial community composition and function of the Temengor Lake metagenomes were observed. The photic water layer of Temengor Lake was dominated by typical freshwater assemblages consisting of Proteobacteria, Actinobacteria, Bacteroidetes, Verrucomicrobia, and Cyanobacteria lineages. On the other hand, the aphotic water featured in addition to Proteobacteria, Bacteroidetes, Verrucomicrobia, and two more abundant bacterial phyla that are typically ubiquitous in anoxic habitats (Chloroflexi and Firmicutes). The aphotic zone of Temengor Lake exhibited genetic potential for nitrogen and sulfur metabolisms for which terminal electron acceptors other than oxygen are used in the reactions. The aphotic water of the disturbed site also showed an overrepresentation of genes associated with the metabolism of carbohydrates, likely driven by the enrichment of nutrient resulting from aquaculture activities at the site. The results presented in this study can serve as a basis for understanding the structure and functional capacity of the microbial communities in the photic and aphotic zones/water layers of tropical man-made lakes.
This study aimed to determine effects of 6-day progressive increase in salinity from 1 per thousand to 15 per thousand on nitrogen metabolism and excretion in the soft-shelled turtle, Pelodiscus sinensis. For turtles exposed to 15 per thousand water on day 6, the plasma osmolality and concentrations of Na+, Cl- and urea increased significantly, which presumably decreased the osmotic loss of water. Simultaneously, there were significant increases in contents of urea, certain free amino acids (FAAs) and water-soluble proteins that were involved in cell volume regulation in various tissues. There was an apparent increase in proteolysis, releasing FAAs as osmolytes. In addition, there might be an increase in catabolism of certain amino acids, producing more ammonia. The excess ammonia was retained as indicated by a significant decrease in the rate of ammonia excretion on day 4 in 15 per thousand water, and a major portion of it was converted to urea. The rate of urea synthesis increased 1.4-fold during the 6-day period, although the capacity of the hepatic ornithine urea cycle remained unchanged. Urea was retained for osmoregulation because there was a significant decrease in urea excretion on day 4. Increased protein degradation and urea synthesis implies greater metabolic demands, and indeed turtles exposed to 15 per thousand water had significantly higher O2 consumption rate than the freshwater (FW) control. When turtles were returned from 15 per thousand water to FW on day 7, there were significant increases in ammonia (probably released through increased amino acid catabolism) and urea excretion, confirming that FAAs and urea were retained for osmoregulatory purposes in brackish water.
Helicobacter pylori colonizes almost half of the human population worldwide. H. pylori strains are genetically diverse, and the specific genotypes are associated with various clinical manifestations including gastric adenocarcinoma, peptic ulcer disease (PUD), and nonulcer dyspepsia (NUD). However, our current knowledge of the H. pylori metabolism is limited. To understand the metabolic differences among H. pylori strains, we investigated four Malaysian H. pylori clinical strains, which had been previously sequenced, and a standard strain, H. pylori J99, at the phenotypic level.
The aeration strategy ranging from intermittent to continuous aeration in the REACT period of moving bed sequencing batch reactor (MBSBR) was evaluated for simultaneous removal of 4-chlorophenol (4-CP) and nitrogen. The results show that the removal rates of 4-CP and ammonium nitrogen (NH(4)(+)-N) increased with the increase of continuous aeration period. In the presence of 4-CP, NH(4)(+)-N removal was mainly by the assimilation process. The removal of NH(4)(+)-N to oxidized nitrogen via oxidation was only observed after 4-CP was completely degraded with sufficient aeration period provided indicating the inhibitory effect of 4-CP on nitrification. As the intermittent aeration strategy would lead to slower 4-CP degradation resulting in the delay of nitrification process, continuous aeration would be the preferred strategy in the simultaneous removal of 4-CP and nitrogen in the MBSBR system.