Pesticide Residue was Affect Bacterial and Fungal Population in the Greenhouse Soil Condition. Bacterial and fungal inhabitants were examined through agriculture soil samples. Survival of microorganism in soil was important to evaluate the mineralize process. In the greenhouse experiment, bacterial and fungal population noticed as essential assessment in soil healing with long lasting pesticide (A-soil) after the soil treated with powdered rice straw as organic matter amendment, toxic degrading bacterial as inoculants, and the both of those mixed treatments. That residue implication also assessed to B-soil (as free pesticide soil) then revised with pesticides, and also treated as followed for A-soil. Potential degradation of indigenous microorganism examined all through as setting of control (B-soil) to recognize of its original population.Those of treatments evaluated through microbial survival along 12 weeks incubation in green house environment. Bacterial and fungal populations was affect to become fluctuated along with incubation period as due to the treatments. Bacterial inhabitant was considerably higher compared to fungal population. Correlations were significantly difference with bacterial (p = 0.6654) as well as fungal (p = 0.9029) population in A-soil evaluated to B-soil habitats since the mixed treatments present to both of the soil (p0.005 = 0.6310). Organic matter alteration and certain microbe input was needed in soil contain pesticide, because at the same time the survival of microbes possibly mineralized organic matter, as well as the pesticide residue.
<p>The objective of this study was to compare the efficiency of three methods of nucleic acids extraction from gram-positive bacteria by evaluating the quantity and purity of DNA extracts. Nucleic acids extraction of gram-positive bacteria is normally hampered by a thick and resistant cell wall. Gram positive bacteria usually have a thick cell wall consisting mainly of many layers of peptidoglycan, which is not easily destroyed. This paper compares the different procedures based on mechanical and enzymatic cell breakage to extract DNA from Rhodoccocus pyridinivorans using GES method, Ultraclean Microbial DNA isolation Kit, and Prepman Microbial DNA isolation kit. DNA extracts were analyzed by agarose gel electrophoresis and UV spectroscopy. Yield and quality of DNA obtained by the GES method were higher than the other methods. Nucleic acids extracts with the highest yield and purity were amplified by Polymerase Chain Reaction (PCR) using various primers targeted on gene encoding nitrilase gene such as BLITF and PNITR, a NH1 and a NH2, ß NH1 and ß NH2, Amd1 and Amd 2. The gene encoding for nitrilase were amplified which was confirmed by sequencing analyses. However, the targeted gene length from the primes was not obtained. Therefore further amplification optimization may be needed.</p><p><br /><strong>Keywords</strong>: extraction, DNA isolation Rhodococcus pyridinovorans, nitrilase gene</p>
Acrylonitrile is a toxic organo-cyanide compound extensively used as solvents and in the manufacture of plastics, polymers, synthetic fibers, resin, dyestuffs, pharmaceuticals and vitamins. Because of its acute neurotoxicity, mutagenicity, carcinogenicity and teratogenicity, discharge of acrylonitrile contained wastewater can lead to serious environmental pollution if is not controlled. Microbial degradation has been considered as a way of removing highly toxic nitriles from industrial waste. In our studies, the biodegradation of acrylonitrile was demonstrated by using whole cells of Corynebacterum sp. D5 isolated from polluted industrial wastewater. Although the bacterium could not utilized the compound as a source of carbon, energy and nitrogen for its growth, Corynebacterium sp. D5 was capable to degrade acrylonitrile (CH2=CH-CN) into acrylamide (CH2=CH-CONH2) and acrylic acid (CH2=CH-OOH). The acrylonitrile degradation took place via a two-steps reaction invoving nitrile-hydratase and amidase. During the degradation, the highest nitril hydratase activity was 4.894 nmol.(min.mg) −1 with the optimum temperature and pH was 25°C and pH 7.0, while the highest amidase acivity was 1,315 nmol.(min.mg) −1, with the optimum temparature and pH was 50°C and pH 6.0, respectively. Besides on acetonitrile as inducer, Corynebacterium sp. D5 was also able to grow on various saturated low molecular weight of nitrile and amide compounds.
This paper describes a efficient screening gene nitrilase from contaminated soil from Lombok gold mine industry. DNA was extracted directly from soil using the soil DNA isolation kit based on enzymatic, chemical and mechanical lysis. The existence of nitrilase gene in soil sample can be identified by nitrilase gene amplification using H1F-H1R primer. BLASTN analysis result revealed that the nitrilase gene fragment which was amplified by H1F-H1R primer has a high homology with Rhodococcus rhodochrous strain tg1-A6 nitrilase gene. These amplification and DNA fragment sequencing results indicated that nitrilase gene existence on soil sample can be identified by metagenomic approach
Bacterial consortium capable of growing and utilizing cyanide as a source of nitrogen were isolated from effluent of gold mining industry. The isolation was conducted using liquid enrichment medium with potassium cyanide and glucose as nitrogen and carbon source, respectively. These consortium could tolerate and were able to grow on KCN at concentration of up to 1000 ppm. Bacterial consortium LP3 were also able to degrade potassium cyanide and ammonium as product of the degradation. The degradation rate was 9,0μM per minute. The cyanide-degrading bacteria found in this consortium were identified as Bacillus, Corynebacterium, and Serratia.Keywords: potassium cyanide, nitrogen source, bacterial consortium LP3,degradation
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