Coagulation/flocculation-based removal of algal-bacterial biomass from piggery wastewater treatment.

Two conventional chemical coagulants (FeCl3 and Fe2(SO4)3) and five commercial polymeric flocculants (Drewfloc 447, Flocudex CS/5000, Flocusol CM/78, Chemifloc CV/300 and Chitosan) were comparatively evaluated for their ability to remove algal-bacterial biomass from the effluent of a photosynthetically oxygenated piggery wastewater biodegradation process. Chlorella sorokiniana, Scenedesmus obliquus, Chlorococcum sp. and a wild type Chlorella, in symbiosis with a bacterial consortium, were used as model algal-bacterial consortia. While the highest biomass removals (66-98%) for the ferric salts were achieved at concentrations of 150-250 mg L(-1), dosages of 25-50 mg L(-1) were required for the polymer flocculants to support comparable removal efficiencies. Process efficiency declined when the polymer flocculant was overdosed. Biomass concentration did not show a significant impact on flocculation within the concentration range tested. The high flocculant requirements herein recorded might be due to the competition of colloidal organic for the flocculants and the stationary phase conditions of biomass.

A comparative evaluation of microalgae for the degradation of piggery wastewater under photosynthetic oxygenation.

Two green microalgae (Scenedesmus obliquus and Chlorella sorokiniana), one cyanobacterium (Spirulina platensis), one euglenophyt (Euglena viridis) and two microalgae consortia were evaluated for their ability to support carbon, nitrogen and phosphorous removal in symbiosis with activated sludge bacteria during the biodegradation of four and eight times diluted piggery wastewater in batch tests. C. sorokiniana and E. viridis were capable of supporting the biodegradation of four and eight times diluted wastewater. On the other hand, while S. obliquus and the consortia isolated from a swine manure stabilization pond were only able to grow in eight times diluted wastewater, S. platensis and the consortium isolated from a high rate algal pond treating swine manure were totally inhibited regardless of the dilution applied. TOC removal efficiencies (RE) ranging from 42% to 55% and NH(4)(+)-RE from 21% to 39% were recorded in the tests exhibiting photosynthetic oxygenation. The similar oxygen production rates exhibited by the tested microalgae under autotrophic conditions (from 116 to 133mgO(2)L(-1)d(-1)) suggested that factors other than the photosynthetic oxygenation potential governed piggery wastewater biodegradation. Microalgal tolerance towards NH(3) was hypothesized as the key selection criterion. Further studies in a continuous algal-bacterial photobioreactor inoculated with C. sorokiniana, S. obliquus and S. platensis showed that C. sorokiniana, the species showing the highest NH(3)-tolerance, rapidly outcompeted the rest of the microalgae during the biodegradation of eight times diluted wastewater, achieving TOC and NH(4)(+)-RE comparable to those recorded in the batch biodegradation tests.

Production of a lipolytic enzyme originating from Bacillus halodurans LBB2 in the methylotrophic yeast Pichia pastoris.

A gene encoding a lipolytic enzyme amplified from the alkaliphilic bacterium Bacillus halodurans LBB2 was cloned into the pPICZalphaB vector and integrated into the genome of the protease deficient yeast strain Pichia pastoris SMD1168H. This previously undescribed enzyme was produced in active form, and cloning in frame with the Saccharomyces cerevisiae secretion signal (alpha-factor) enabled extracellular accumulation of correctly processed enzyme, with an apparent molecular mass of 30 kDa. In shake-flask cultivations, very low production levels were obtained, but these were significantly improved by use of a "batch-induced" cultivation technique which allowed a maximum enzyme activity of 14,000 U/l using p-nitrophenyl butyrate (C-4) as a substrate and a final extracellular lipolytic enzyme concentration of approximately 0.2 g/l. Partial characterization of the produced enzyme (at pH 9) revealed a preference for the short-chain ester (C-4) and significant but lower activity towards medium (C5-C6) and long (C16 and C18) fatty acid chain-length esters. In addition, the enzyme exhibited true lipase activity (7,300 U/l) using olive oil as substrate and significant levels of phospholipase activity (6,400 U/l) by use of a phosphatidylcholine substrate, but no lysophospholipase activity was detected using a lysophosphatidylcholine substrate.

Bacillus bogoriensis sp. nov., a novel alkaliphilic, halotolerant bacterium isolated from a Kenyan soda lake.

Strain LBB3(T) isolated from Bogoria soda lake in Kenya is an alkaliphilic, Gram-positive, strictly aerobic, non-motile, spore-forming bacterium. It was identified as a member of the genus Bacillus on the basis of phenotypic and phylogenetic analyses. The organism grows optimally at 37 degrees C and pH 10. The G+C content of the genomic DNA is 37.5 mol%. 16S rRNA gene sequence analysis showed 95 and 96 % sequence similarity with Bacillus pseudofirmus (DSM 8715(T)) and Bacillus alcalophilus (DSM 485(T)), respectively. Furthermore, DNA-DNA hybridization against these two Bacillus species showed 39.0 and 55.5 % similarity, respectively. Based on our observations, strain LBB3(T) is proposed to represent a novel species of the genus Bacillus, for which the name Bacillus bogoriensis sp. nov. is proposed. The type strain of B. bogoriensis is LBB3(T) (=ATCC BAA-922(T)=LMG 22234(T)).

Lipase-producing microorganisms from a Kenyan alkaline soda lake.

Lipolytic enzyme production of 150 isolated strains from samples of Lake Bogoria (Kenya) was examined. Among these, fifteen isolates were selected on the basis of their lipolytic activities and subjected to morphological and 16S rRNA gene sequencing analyses for their identification. All the microorganisms have been selected under culture conditions with pH ranges between 7-10 and temperatures of 37-55 degrees C. Most of them showed optimal growth at 37 degrees C and tolerated salinity up to 10% (w/v). Ten of the isolates were Gram-negative, nine of which were closely related to the Pseudomonas cluster and one to the Halomonas cluster sharing high similarity profile with Halomonas desiderata. The remaining Gram-positive isolates were closely related to the Bacillus cluster, and were grouped with Bacillus halodurans, Bacillus alcalophilus and Bacillus licheniformis. Four members of the Bacillus cluster and the Halomonas sp. produced lipolytic activity under alkaline conditions, while others did so at neutral pH values.