Molybdenum is very toxic to agricultural animals. Mo-reducing bacterium can be used to immobilize soluble molybdenum to insoluble forms, reducing its toxicity in the process. In this work the isolation of a novel molybdate-reducing Gram positive bacterium tentatively identified as Bacillus sp. strain A.rzi from a metal-contaminated soil is reported. The cellular reduction of molybdate to molybdenum blue occurred optimally at 4 mM phosphate, using 1% (w/v) glucose, 50 mM molybdate, between 28 and 30°C and at pH 7.3. The spectrum of the Mo-blue product showed a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of bacterial electron transport system (ETS) such as rotenone, sodium azide, antimycin A, and potassium cyanide could not inhibit the molybdenum-reducing activity. At 0.1 mM, mercury, copper, cadmium, arsenic, lead, chromium, cobalt, and zinc showed strong inhibition on molybdate reduction by crude enzyme. The best model that fitted the experimental data well was Luong followed by Haldane and Monod. The calculated value for Luong's constants p max, K s, S m, and n was 5.88 μmole Mo-blue hr−1, 70.36 mM, 108.22 mM, and 0.74, respectively. The characteristics of this bacterium make it an ideal tool for bioremediation of molybdenum pollution.
Phenol is an important pollutant widely discharged as a component of hydrocarbon fuels, but its degradation in cold regions is a great challenge due to the harsh environmental conditions. To date, there is little information available concerning the biodegradation of phenol by indigenous Antarctic bacteria. This study addresses the isolation of three phenol-degrading bacterial strains from King George Island, Antarctica. Based on preliminary screening, three isolates (AQ5-05, AQ5-06 and AQ5-07) capable of completely degrading 0.5 g/L phenol within 120 h at 10 °C were selected for detailed study. Two were identified as Arthrobacter spp., and one Rhodococcus sp., based on 16S rRNA sequences. All strains were non-motile, Gram positive, oxidase negative and catalase positive. A study on the effects of parameters including temperature, pH, salinity and nitrogen source was conducted to optimise the conditions for phenol degradation. This revealed that the three isolates were psychrotolerant with the optimum temperature for phenol degradation between 10 and 15 °C. This study suggests the potential use of cold-adapted bacteria in the bioremediation of phenol over a wide range of low temperatures.
Acetylcholinesterase (AChE) generally known to be sensitive toward insecticides but its sensitivity toward heavy metals was least reported. Herein, a sensitive assay for heavy metals has been pursued using AChE in a rapid and economic manner. The AChE from a mudskipper, Periophthalmodon schlosseri has been found to be sensitive toward copper [ mercury [ chromium [ and arsenic ions at the sub parts per million levels. Field trial works showed that the assay was applicable in detecting heavy metals pollution from effluents of industrial sites at near real time and verified using ICP-OES and Flow Injection Mercury System (FIMS 400). Furthermore, hierarchical cluster analyses of inhibition profiles were performed, revealing a comparable capability of the AChE compared to the gold standard of Microtox™ method.
BackgroundBiodegradation of hydrocarbons in Antarctic soil has been reported to be achieved through the utilisation of indigenous cold-adapted microorganisms. Although numerous bacteria isolated from hydrocarbon-contaminated sites in Antarctica were able to demonstrate promising outcomes in utilising hydrocarbon components as their energy source, reports on the utilisation of hydrocarbons by strains isolated from pristine Antarctic soil are scarce. In the present work, two psychrotolerant strains isolated from Antarctic pristine soil with the competency to utilise diesel fuel as the sole carbon source were identified and optimised through conventional and response surface method.ResultsTwo potent hydrocarbon-degraders (ADL15 and ADL36) were identified via partial 16S rRNA gene sequence analysis, and revealed to be closely related to the genus Pseudomonas and Rhodococcus sp., respectively. Factors affecting diesel degradation such as temperature, hydrocarbon concentration, pH and salt tolerance were studied. Although strain ADL36 was able to withstand a higher concentration of diesel than strain ADL15, both strains showed similar optimal condition for the cell’s growth at pH 7.0 and 1.0% (w/v) NaCl at the conventional ‘one-factor-at-a-time’ level. Both strains were observed to be psychrotrophs with optimal temperatures of 20 °C. Qualitative and quantitative analysis were performed with a gas chromatograph equipped with a flame ionisation detector to measure the reduction of n-alkane components in diesel. In the pre-screening medium, strain ADL36 showed 83.75% of n-dodecane mineralisation while the reduction of n-dodecane by strain ADL15 was merely at 22.39%. The optimised condition for n-dodecane mineralisation predicted through response surface methodology enhanced the reduction of n-dodecane to 99.89 and 38.32% for strain ADL36 and strain ADL15, respectively.ConclusionsStrain ADL36 proves to be a better candidate for bioaugmentation operations on sites contaminated with aliphatic hydrocarbons especially in the Antarctic and other cold regions. The results obtained throughout strongly supports the use of RSM for medium optimisation.
The status report on metal pollution in tropical estuaries and coastal waters is important to understand potential environmental health hazards. Detailed baseline measurements were made on physicochemical parameters (pH, temperature, redox potential, electrical conductivity, salinity, dissolved oxygen, total dissolved solid), major ions (Na, Ca, Mg, K, HCO3, Cl, SO4 and NO3) and metals concentrations ((27)Al, (75)As, (138)Ba, (9)Be, (111)Cd, (59)Co, (63)Cu, (52)Cr, (57)Fe, (55)Mn, (60)Ni, (208)Pb, (80)Se, (66)Zn) at estuaries and coastal waters along the Straits of Malacca. Principal component analysis (PCA) was employed to reveal potential pollution sources. Seven principal components were extracted with relation to pollution contribution from minerals-related parameters, natural and anthropogenic sources. The output from this study will generate a profound understanding on the metal pollution status and pollution risk of the estuaries and coastal system.
Luminescence-based assays for toxicants such as Microtox, ToxAlert, and Biotox have been used extensively worldwide. However, the use of these assays in near real time conditions is limited due to nonoptimal assay temperature for the tropical climate. An isolate that exhibits a high luminescence activity in a broad range of temperatures was successfully isolated from the mackerel, Rastrelliger kanagurta. This isolate was tentatively identified as Photobacterium sp. strain MIE, based on partial 16S rDNA molecular phylogeny. Optimum conditions that support high bioluminescence activity occurred between 24 and 30°C, with pH 5.5 to 7.5, 10 to 20 g/L of sodium chloride, 30 to 50 g/L of tryptone, and 4 g/L of glycerol as the carbon source. Assessment of near real time capability of this bacterial system, Xenoassay light to monitor heavy metals from a contaminated river running through the Juru River Basin shows near real time capability with assaying time of less than 30 minutes per samples. Samples returned to the lab were tested with a standard Microtox assay using Vibrio fishceri. Similar results were obtained to Xenoassay light that show temporal variation of copper concentration. Thus, this strain is suitable for near real time river monitoring of toxicants especially in the tropics.
Caffeine is an important naturally occurring compound that can be degraded by bacteria. Excessive caffeine consumption is known to have some adverse effects. We isolated a new bacterium from agriculture soil. The bacterium was tested for its ability to utilise caffeine as the sole carbon and nitrogen source. The isolate was Gram-negative and was identified as Leifsonia sp. strain SIU based on 16S rRNA gene sequencing. It showed 97.16 % of 0.3 g/L caffeine degradation in 48 h when caffeine was used as a sole carbon and nitrogen source. The bacterial growth and degradation at 0.3 g/L caffeine concentration occurred optimally, using 5 g/L sucrose, 0.4 g/L ammonium chloride, at a temperature between 25 and 30°C and pH of 6.0-7.0. The Luong model best describes the kinetics of the strain growth. The values for the maximum specific growth rate (μ max), the Monod half saturation constant (K S), the maximum substrate inhibitory concentration and n are 0.049 h−1, 0.0021 mg/L, 25.0 g/L and 1.562, respectively. These bacterial features make it an ultimate means for caffeine bioremediation. This is the first report of caffeine degradation by Leifsonia sp. strain SIU.
Rhodococci are renowned for their great metabolic repertoire partly because of their numerous putative pathways for large number of specialized metabolites such as biosurfactant. Screening and genome-based assessment for the capacity to produce surface-active molecules was conducted on Rhodococcus sp. ADL36, a diesel-degrading Antarctic bacterium. The strain showed a positive bacterial adhesion to hydrocarbon (BATH) assay, drop collapse test, oil displacement activity, microplate assay, maximal emulsification index at 45% and ability to reduce water surface tension to < 30 mN/m. The evaluation of the cell-free supernatant demonstrated its high stability across the temperature, pH and salinity gradient although no correlation was found between the surface and emulsification activity. Based on the positive relationship between the assessment of macromolecules content and infrared analysis, the extracted biosurfactant synthesized was classified as a lipopeptide. Prediction of the secondary metabolites in the non-ribosomal peptide synthetase (NRPS) clusters suggested the likelihood of the surface-active lipopeptide production in the strain’s genomic data. This is the third report of surface-active lipopeptide producers from this phylotype and the first from the polar region. The lipopeptide synthesized by ADL36 has the prospect to be an Antarctic remediation tool while furnishing a distinctive natural product for biotechnological application and research.
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