Category Archives: 2013 Vol 25 Dec

Zero Discharge Treatment Technology of Palm Oil Mill Effluent

Palm oil processing operation is invariably accompanied by palm oil mill effluent (POME) considered to be an environmental pollutant. While anaerobic digestion and the present tertiary treatment technologies of POME are able to meet the current regulatory effluent discharge requirement of biological oxygen demand (BOD) 100 mg litre-1 – the current limit set by the Department of Environment (DOE) – the existing technologies are unable to consistently meet the proposed stringent BOD regulatory requirement of 20 mg litre-1 to be imposed by the DOE. This article investigates the possibility of integrating several bioprocesses for POME treatment at the Malaysian palm oil mills by transforming the POME into several high valueadded products; with BOD 20 mg litre-1 attainable at its final discharge. This integrated approach has the potential of achieving zero-effluent discharge along with the production of biogas, biofertiliser and recycled water, in treating industrial wastewater to reduce pollution.

Leaching and Persistence of Benomyl in an Oil Palm Nursery

A field trial on the leaching and persistence of benomyl in an oil palm nursery agro ecosystem was conducted at Labu, Negeri Sembilan, Malaysia. Benomyl was analysed as its degradation product carbendazims because benomyl is a very unstable compound and easily degrades in organic solvents and aqueous solutions. Therefore, the quantification of benomyl was expressed as carbendazim. Experimental plots in the nursery were sprayed with the fungicide benomyl, while the control plot was left untreated. The fungicide was sprayed using a knapsack sprayer at the recommended manufacturer’s dosage as well as at double the recommended dosage. Composite soil samples were collected from each replicate plot at depths of
0-10 cm, 10-20 cm, 20-30 cm, 30-40 cm and 40-50 cm. Sampling of soils were taken at intervals of -1, 0, 1, 2, 3, 5, 7, 14, 30, 60 and 90 day/days after fungicide treatment. Analytical methods for the determination of carbendazim in soils were developed and good recoveries of over 80% with small standard deviations were obtained from soil samples. Carbendazim residues were detected until Day 1 after treatment. The carbendazim residues were found at a depth of 0-50 cm for both normal as well as double recommended dosages. The objective of this study was to evaluate the mobility and persistence of the fungicide benomyl in an oil palm nursery.

Preparation of Polyglycerol from Palm-Biodiesel Crude Glycerin

The demand for green energy derived from plant feedstock has been the determining factor for the abundant supply of glycerol. Concerted efforts have been made to expand the current scope of glycerol application. This study describes the rapid polymerisation of crude glycerol directly obtained from the biodiesel process to produce polyglycerol via microwave heating technology. Crude glycerol used in this study was obtained from four different biodiesel pilot plants. Commercially pure glycerol was used as the control in all the parameters studied. The highest percent yield of polyglycerol obtained was 94.94% when heated at 250oC for 60 min under microwave irradiation. Crude glycerol that gave the highest polyglycerol percentage was found to contain the highest percent of soap (12.5%). It was anticipated that a high conversion of glycerol was due to the soap contained in the crude glycerol. This study showed that biodiesel-based crude glycerol with appropriate soap content could be used directly as a raw material in polyglycerol production.

Identification of Differentially Expressed Proteins in Oil Palm Seedlings Artificially Infected With Ganoderma: A Proteomics Approach

Basal stem rot disease in oil palm is caused by the fungus, Ganoderma boninense. Proteomics approaches using two-dimensional gel electrophoresis (2-DE) were adopted to identify proteins from oil palm that were differentially expressed in response to the fungus. Total proteins were extracted from oil palm roots after Day 3 and Day 7 post-infection and resolved in 2-DE gels. Differentially expressed protein spots between the infected and uninfected palms were determined using PD-Quest Software. Seven protein spots were differentially expressed in the infected root at Day 3 post-infection. At Day 7, the number of differentially expressed protein spots increased to 25, with a molecular weight less than 50 kDa. All protein spots were analysed by MALDI TOF/TOF mass spectrometry. Twelve candidate proteins were identified by a comparison of differentially expressed proteins at Day 7 with the database. One of the candidate proteins is glucanase, which is widely known to induce during fungal response in other plants. Other candidate proteins that were identified were also involved in defence and stress responses in other plants, suggesting that these proteins play a significant role in oil palm root pathogenicity response towards G. boninense infection

Representational difference analysis (RDA) for the identification of DNA markerS associated with tissue culture amenity in oil palm

Tissue culture is a promising technology for mass propagation of high yielding oil palm. However, the large scale production of clonal palm is still not optimal. The application of DNA marker for early diagnosis of high quality ortets (selected oil palm tree for cloning) would greatly help in reducing the production cost. Representational difference analysis (RDA) was used to identify genomic differences between the oil palm explant tissues of the same genotypes but producing different somatic embryogenesis rates in culture. The putative RDA products isolated were cloned, sequenced and verified. Among the products isolated, C1-20 and C14-19 from forward and reverse RDA respectively, possess a single nucleotide change when aligned with the equivalent DNA region of the subtracted tissues. However, further verification is needed to confirm the discriminating effects of these products on the fecundity of the selected ortets.

The Effects Of Trichoderma In Surface Mulches Supplemented With Conidial Drenches In The Disease Development Of Ganoderma Basal Stem Rot In Oil Palm

Biocontrol efficiency of Trichoderma in surface mulches together with conidial drenches on disease suppression of basal stem rot (BSR) caused by Ganoderma boninense in oil palm seedlings was investigated. Surface mulches used were from palm press fibre (PPF). Two isolates of Trichoderma (T. asperellum T9 and T. virens T29) were selected based on the in vitro antagonistic assessment via dual culture followed by poison food agar test. In vivo results indicated disease development was lowest in plants treated with Trichoderma T29 and Trichoderma T9 compared to plants treated with mixed inocula of Trichoderma (T9 and T29), PPF alone and water. Disease development in the plants was assessed using disease severity (DS) and subjected to further analysis with area under the disease progression curve (AUDPC) and epidemic rate (RL). Disease development was delayed by eight weeks in plants treated with Trichoderma T29 and Trichoderma T9 compared to control treatment. The study also suggests that the potential use of PPF, a biowaste product of palm oil mill as surface mulch, enriched with single Trichoderma species (T9/T29) effectively delayed disease onset in oil palm seedlings.

Reversed-Phase High-Performance Liquid Chromatography Analysis of 1,3- and 1,2(2,3)-Positional Isomers of Palm-Based Diacylglycerols

Reversed-phase high-performance liquid chromatography method using charged aerosol detector was developed for separation of 1,3- and 1,2(2,3) positional isomers of palm oil- and palm kernel oil-based diacylglycerols (PO-DAG and PKO-DAG, respectively) with different equivalent carbon numbers
(ECN) and without the need of sample derivatisation. In this method, step-wise gradient of acetone and acetonitrile was used and a total retention time of 28 min was attained. Identification of PKO- and PODAG molecular species was accomplished using synthetic DAG standards. Completeness of separation as well as identification of PKO- and PO-DAG molecular species including positional isomers with different ECN values were verified where similar elution patterns as well as the same number of identified peaks were observed for the chromatograms of PKO-DAG and PKO-DAG standards, as well as the PO-DAG and PO-DAG standards. Among the 1,3-DAG species and/or 1,2-DAG species, as ECN value of their fatty acid constituents increased, their corresponding retention time always increased. However, among the PKO- as well as PO-based synthetic DAG with the same ECN values, 1,3-DAG were always found to elute earlier than the respective 1,2-DAG. Furthermore, some exceptional examples were observed among the PKO- as well as PO-based synthetic DAG standards with the different ECN values where a few of 1,3-DAG with the relatively higher ECN values eluted earlier than 1,2-DAG with relatively lower ECN values. The last two observations evidence the defect of ECN-based prediction for identification of DAG positional isomers with the same ECN as well as for different ECN values.

Ammonium Salts of Palm Oil Fatty Acids

Palm oil acid fraction-based monoethanolamine (MEA), diethanolamine (DEA), triethanolamine (TEA), methylaminoethanol (MAE), piperidine (PP), morpholine (Mr), diethylamine (DEtA), triethylamine (TEtA), ethylenediamine (EDA), diethylenetriamine (DETA) and polyethylenepolyamine (PEPA) salts were synthesised. By stalagmometric method it was established that the synthesised salts are highly surfaceactive. DEA and TEA-based salts reduce the interface tension at the air-water surface from 72.0 mN m-1 down to 25.7 and 24.2 mN m-1 (25°C). Using surface tension isotherms of the salts their colloidal-chemical parameters were calculated. An increase in the number of ethylol groups in the surfactant composition decreases critical micelle concentration (CMC) and area per molecule at the interface (Amin) and increases effectiveness of surface tension reduction (pCMC) and surface excess concentration (Gmax). It was revealed that DETA-based salt has a higher petroleum-collecting ability than the other salts.

Determination of Life Cycle Inventory and Greenhouse Gas Emissions for a Selected Oil Palm Nursery in Malaysia: A Case Study

A case study on a gate-to-gate life cycle assessment (LCA) starting from the delivery of germinated seeds to the production of oil palm seedling in an oil palm nursery was carried out. The oil palm nursery, located in Selangor and licensed by the Malaysian Palm Oil Board (MPOB) was selected for this study. One oil palm seedling was the defined functional unit. The environmental impact from activities in the nursery were determined from the input and output into and from the processing steps in the nursery. The LCA study was conducted in accordance with the procedural framework ISO 14040 – 14044 series of the International Organisation of Standardisation (ISO). The purpose of the study was to obtain a life cycle inventory (LCI) of input and output associated with the production of one oil palm seedling and to provide a comprehensive picture of the environmental impact associated with production of one oil palm seedling in the nursery, by applying the LCA methodology. Inventory data collection consists of the input of raw materials and energy such as polybags, fertilisers, pesticides, diesel, pipes and petrol for transportation and irrigation. Real time measurement of gas and greenhouse gas (GHG) emissions from the pump for irrigation in the nursery was also carried out. The results show that the LCA methodology is suitable for assessing the environmental impact associated with oil palm seedling production.

Optimisation of Enzymatic Sludge Palm Oil Recovery from Palm Oil Mill Effluent using Response Surface Methodology

Various operational variables affecting the extraction of sludge palm oil (SPO) from palm oil mill effluent (POME) by enzymatic treatment were investigated. The enzyme used in the study was cellulase from Trichoderma reseei, commercially known as Celluclast 1.5L FG (Novozymes). A software application of the Central Composite Face-centred Design (CCFD) known as Response Surface Methodology (RSM) was used to study the effects of independent variables generated by the Design Expert Software, namely incubation time, X1 (12.73-147.27 hr), enzyme concentration, X2 (0.08%-0.92%) and rotation speed, X3 (65.91-234.09 rpm) on the oil recovered from POME. In this study, a second-order polynomial regression model was used to interpret the experimental data with a coefficient of determination (R2) value of 0.852. From the RSM-generated model, the optimum conditions for extraction of oil from POME were identified to be at an enzyme concentration value of 0.25% in 81.46 hr reaction time, with a rotation speed of 111.05 rpm. Predicted oil yield was 93.56% while the experimental oil yield was 81.95% as revealed by the One Sample T-Test of confirmatory studies.

Analysis of Oil Palm Clones, their Suspension Calli and Regenerants via Flow Cytometry (FCM) and rDNA-fluorescence in situ Hybridisation (rDNA-FISH)

Clonal propagation of palms with good traits is desirable for the oil palm industry. In this study, flow cytometry (FCM) and 18S-25S ribosomal DNA-fluorescence in situ hybridisation (rDNA-FISH) were utilised to analyse genetic variation in adult clonal palms, their respective suspension cultures and regenerant plantlets. The genome sizes estimated by FCM for the four adult clonal palms (using leaf samples from Frond-1) varied from 2C=2.59±0.19 pg to 2.91±0.14 pg and for their respective regenerants, the genome size varied from 2C=2.14±0.21 pg to 3.05±0.11 pg. The genome size of oil palm suspension cultures could not be analysed by FCM due to the low nuclei population, which was less than 1000. The rDNA-FISH analysis showed two hybridisation signals on interphase nuclei of suspension culture calli and regenerant plantlets, hence indicating the diploid ploidy level. Adult clonal palms with their suspension culture calli and regenerants therefore showed a similar ploidy level. However, the measurement of genome size was found to vary between the adult clonal palms and their regenerants

First Report: Isolation of Endophytic Trichoderma from Oil Palm (Elaeis guineensis Jacq.) and Their in vitro Antagonistic Assessment on Ganoderma boninense

The isolation of endophytic Trichoderma from different tissues viz. leaf, rachis, stem and root of oil palm (Elaeis guineensis Jacq.) was investigated in this study. A total of six palms were sampled from two plantation plots and it was found that Trichoderma isolates were specifically colonising the stems and roots of oil palm and none were found in the leaves and rachis. Trichoderma selective media E (TME) was used for isolation. Identification of the isolates was further established using slide cultures. A total of 40 isolates were obtained with 10 from stem and 30 from root samples of oil palm. All isolates were subjected to dual culture assay; a preliminary assay to screen mycoparatisms potential through antagonistic activity against Ganoderma boninense. Subsequently, seven potential Trichoderma isolates were tested for their antibiosis properties through poison food agar assay. This is the first report of endophytic Trichoderma isolated from oil palm with biocontrol potential against G. boninense.