Ecotoxicological assessment of palm oil mill effluent final discharge by zebrafish (Danio rerio) embryonic assay.

Most palm oil mills adopted conventional ponding system, including anaerobic, aerobic, facultative and algae ponds, for the treatment of palm oil mill effluent (POME). Only a few mills installed a bio-polishing plant to treat POME further before its final discharge. The present study aims to determine the quality and toxicity levels of POME final discharge from three different mills by using conventional chemical analyses and fish (Danio rerio) embryo toxicity (FET) test. The effluent derived from mill A which installed with a bio-polishing plant had lower values of BOD, COD and TSS at 45 mg/L, 104 mg/L, and 27 mg/L, respectively. Only mill A nearly met the industrial effluent discharge standard for BOD. In FET test, effluent from mill A recorded low lethality and most of the embryos were malformed after hatching (half-maximal effective concentration (EC50) = 20%). The highest toxicity was observed from the effluent of mill B and all embryos were coagulated after 24 h in samples greater than 75% of effluent (38% of half-maximal lethal concentration (LC50) at 96 h). The embryos in the effluent from mill C recorded high mortality after hatching, and the survivors were malformed after 96 h exposure (LC50 = 26%). Elemental analysis of POME final discharge samples showed Cu, Zn, and Fe concentrations were in the range of 0.10-0.32 mg/L, 0.01-0.99 mg/L, and 0.94-4.54 mg/L, respectively and all values were below the effluent permissible discharge limits. However, the present study found these metals inhibited D. rerio embryonic development at 0.12 mg/L of Cu, and 4.9 mg/L of Fe for 96 h-EC50. The present study found that bio-polishing plant installed in mill A effectively removing pollutants especially BOD and the FET test was a useful method to monitor quality and toxicity of the POME final discharge samples.

Toxicity identification and evaluation of palm oil mill effluent and its effects on the planktonic crustacean Daphnia magna.

Palm oil mill effluent (POME) contains complex and highly biodegradable organic matters so discharging it without appropriate treatment may lead to environmental problems. POME final discharge quality is normally determined based on conventional chemical detection such as by biological oxygen demand (BOD) and chemical oxygen demand (COD). The novelty of the present study is that the toxicity effects of the POME final discharge samples were evaluated based on whole effluent toxicity (WET) and toxicity identification evaluation (TIE) tests using Daphnia magna. The toxicity unit (TU) values were recorded to be in the range from TU = 1.1-11 obtained from WET, and the TIE manipulation tests suggested that a substantial amount of toxic compounds was contained in the POME final discharge. Phenol, 2,6-bis (1,1-dimethylethyl)- and heavy metals such as Cu and Zn were detected in all the effluents and were recognized as being the main toxicants in the POME final discharge. GC/MS analyses also successfully identified cyclic volatile methyl siloxanes; cyclotetrasiloxane, octamethyl- (D4), cyclopentasiloxane, decamethyl- (D5), cyclohexasiloxane, dodecamethyl- (D6). D4 was detected at 0.0148-0.0357 mg/L, which could be potentially toxic. The palm oil industry used only water in the form of steam to process the fruits, and the presence of these compounds might be derived from the detergents and grease used in palm oil mill cleaning and maintenance operations. An appropriate treatment process is thus required to eliminate these toxicants from the POME final discharge. It is recommended that two approaches, chemical-based monitoring as well as biological toxicity-based monitoring, should be utilized for achieving an acceptable quality of POME final discharge in the future.

Four products from Escherichia coli pseudogenes increase hydrogen production.

Pseudogenes are considered to be nonfunctional genes that lack a physiological role. By screening 3985 Escherichia coli mutants using chemochromic membranes, we found four pseudogenes involved in hydrogen metabolism. Knockouts of pseudogenes ydfW and ypdJ had a defective hydrogen phenotype on glucose and formate, respectively. Also, the knockout of pseudogene yqiG formed hydrogen from formate but not from glucose. For the yqiG mutant, 100% hydrogen recovery was obtained by the complementation of YqiG via a plasmid. The knockout of pseudogene ylcE showed hydrogen deficiency in minimal media which suggested that the role of YlcE is associated with cell growth. Hence, the products of these four pseudogenes play an important physiological role in hydrogen production in E. coli.

HCHL expression in hairy roots of Beta vulgaris yields a high accumulation of p-hydroxybenzoic acid (pHBA) glucose ester, and linkage of pHBA into cell walls.

As part of a study to explore the potential for new or modified bio-product formation, Beta vulgaris (sugar beet) has been genetically modified to express in root-organ culture a bacterial gene of phenylpropanoid catabolism. The HCHL gene, encoding p-hydroxycinnamoyl-CoA hydratase/lyase, was introduced into B. vulgaris under the control of a CaMV 35S promoter, using Agrobacterium rhizogenes LBA 9402. Hairy root clones expressing the HCHL gene, together with non-expressing clones, were analysed and revealed that one expression-positive clone accumulated the glucose ester of p-hydroxybenzoic acid (pHBA) at about 14% on a dry weight basis. This is the best yield achieved in plant systems so far. Determination of cell-wall components liberated by alkaline hydrolysis confirmed that the ratio of pHBA to ferulic acid was considerably higher in the HCHL-expressing clones, whereas only ferulic acid was detected in a non-expressing clone. The change in cell-wall components also resulted in a decrease in tensile strength in the HCHL-expressing clones.