Biotreatment of industrial olive washing water by synergetic association of microalgal-bacterial consortia in a photobioreactor.

This study presents an effective technology for the olive processing industry to remediate olive washing water. A 14.5-L enclosed tubular photobioreactor was inoculated with a stable microalgal-bacterial consortium obtained by screening strains well adapted to olive washing water. The capacity of an enclosed tubular photobioreactor to remove toxic compounds was evaluated under photosynthesis conditions and without any external supply of oxygen. The results showed that the dominant green microalgae Scenedesmus obliquus, Chlorella vulgaris and the cyanobacteria Anabaena sp. and bacteria present in olive washing water (i.e. Pantoea agglomerans and Raoultella terrigena) formed a synergistic association that was resistant to toxic pollutants present in the effluent and during the initial biodegradation process, which resulted in the breakdown of the pollutant. Total phenolic compounds, COD, BOD5, turbidity and colour removals of 90.3 ± 11.4, 80.7 ± 9.7, 97.8 ± 12.7, 82.9 ± 8.4 and 83.3 ± 10.4 %, respectively, were recorded in the photobioreactor at 3 days of hydraulic retention time. Graphical abstract Biotreatment of industrial olive washing water by synergetic association of microalgal-bacterial consortia in a photobioreactor.

The ratio of metabolically active versus total Mycolata populations triggers foaming in a membrane bioreactor.

The abundance of total and metabolically active populations of Mycolata was evaluated in a full-scale membrane bioreactor (MBR) experiencing seasonal foaming, using quantitative PCR (qPCR) and retrotranscribed qPCR (RT-qPCR) targeting the 16S rRNA gene sequence. While the abundance of total Mycolata remained stable (10(10) copies of 16S rRNA genes/L activated sludge) throughout four different experimental phases, significant variations (up to one order of magnitude) were observed when the 16S rRNA was targeted. The highest ratios of metabolically active versus total Mycolata populations were observed in samples of two experimental phases when foaming was experienced in the MBR. Non-metric multidimensional scaling and BIO-ENV analyses demonstrated that this ratio was positively correlated to the concentrations of substrates in the influent water, F/M ratio, and pH, and negatively correlated to temperature and solids retention time. It the first time that the ratio of metabolically active versus total Mycolata is found to be a key parameter triggering foaming in the MBR; thus, we propose it as a candidate predictive tool.

Isolation and metagenomic characterization of bacteria associated with calcium carbonate and struvite precipitation in a pure moving bed biofilm reactor-membrane bioreactor.

A bench-scale pure moving bed bioreactor-membrane bioreactor (MBBR-MBR) used for the treatment of urban wastewater was analyzed for the identification of bacterial strains with the potential capacity for calcium carbonate and struvite biomineral formation. Isolation of mineral-forming strains on calcium carbonate and struvite media revealed six major colonies with a carbonate or struvite precipitation capacity in the biofouling on the membrane surface and showed that heterotrophic bacteria with the ability to precipitate calcium carbonate and struvite constituted ~7.5% of the total platable bacteria. These belonged to the genera Lysinibacillus, Trichococcus, Comamomas and Bacillus. Pyrosequencing analysis of the microbial communities in the suspended cells and membrane biofouling showed a high degree of similarity in all the samples collected with respect to bacterial assemblage. The study of operational taxonomic units (OTUs) identified through pyrosequencing suggested that ~21% of the total bacterial community identified in the biofouling could potentially form calcium carbonate or struvite crystals in the pure MBBR-MBR system used for the treatment of urban wastewater.

Linking operation parameters and environmental variables to population dynamics of Mycolata in a membrane bioreactor.

The community structure and population dynamics of Mycolata were monitored in a full-scale membrane bioreactor during four experimental phases under changing operating and environmental conditions, by means of temperature-gradient gel electrophoresis of partial 16S-rRNA genes amplified from community DNA and RNA templates (total and active populations). Non-metric multidimensional scaling and BIO-ENV analyses demonstrated that population dynamics were mostly explained (30-32%) by changes in the input of nutrients in the influent water and the accumulation of biomass in the bioreactors, while the influence of hydraulic and solid retention times, temperature and F/M ratio was minor. Significant correlations were observed between particular Mycolata phylotypes and one or more variables, contributing information for the prediction of their abundance and activity under changing conditions. Fingerprinting and multivariate analyses demonstrated that two foaming episodes, recorded at temperatures <20°C, were connected to the increase of the relative abundance of Mycolata unrelated to Gordonia amarae.

Effect of ciprofloxacin antibiotic on the partial-nitritation process and bacterial community structure of a submerged biofilter.

A partial-nitritation bench-scale submerged biofilter was used for the treatment of synthetic wastewater containing a high concentration of ammonium in order to study the influence of the antibiotic ciprofloxacin on the partial-nitritation process and biodiversity of the bacterial community structure. The influence of ciprofloxacin was evaluated in four partial-nitritation bioreactors working in parallel, which received sterile synthetic wastewater amended with 350 ng/L of ciprofloxacin (Experiment 1), synthetic wastewater without ciprofloxacin (Experiment 2), synthetic wastewater amended with 100 ng/L of ciprofloxacin (Experiment 3) and synthetic wastewater amended with 350 ng/L of ciprofloxacin (Experiment 4). The concentration of 100 ng/L of antibiotics demonstrated that the partial-nitritation process, microbial biomass and bacterial structure generated by tag-pyrosequencing adapted progressively to the conditions in the bioreactor. However, high concentrations of ciprofloxacin (350 ng/L) induced a decay of the partial-nitritation process, while the total microbial biomass was increased. Within the same experiment, the bacterial community experienced sequential shifts with a clear reduction of the ammonium oxidation bacteria (AOB) and an evident increase of Commamonas sp., which have been previously reported to be ciprofloxacin-resistant. Our study suggests the need for careful monitoring of the concentration of antibiotics such as ciprofloxacin in partial-nitritation bioreactors, in order to choose and maintain the most appropriate conditions for the proper operation of the system.

Performance of bench-scale membrane bioreactor under real work conditions using pure oxygen: viscosity and oxygen transfer analysis.

Pure oxygen to supply the aerobic condition was used in the performance of a bench-scale submerged membrane bioreactor (MBR). The pilot plant was located in the wastewater treatment plant of the city of Granada (Spain) and the experimental work was divided into two stages (Unsteady state and steady state conditions). Operation parameters (MLSS, MLVSS and dissolved oxygen concentration) and physical characteristics (temperature, conductivity, pH, COD and BOD(5)) were daily monitored. The results showed the capacity of the MBR systems to remove organic material under a hydraulic retention time of 18.46 h and sludge retention time of 18.6 days. Therefore, Viscosity of the sludge and alphakLa-factor of the aeration, were determinate in the steady stage condition to understand the behavior of the system when pure oxygen has been used to supply the aerobic conditions of the MBR system showed an alpha-factor of 0.238 when the viscosity of the system was 4.04 Cp.

Identification of bacteria isolated from an oligotrophic lake with pesticide removal capacities.

We studied the growth and capacities for pesticides removal of bacterial strains isolated from the Laguna Grande, an oligotrophic lake at the South of Spain (Archidona, Málaga). Strains were isolated from water samples amended with 10 and 50 microg/ml of nine pesticides: organochlorinated insecticides (aldrin and lindane), organophosphorous insecticides (dimetoate, methyl-parathion and methidation), s-triazine herbicides (simazine and atrazine), fungicide (captan) and diflubenzuron (1-(-4-chlorophenyl)-3-(2,6-difluorobenzoyl urea), a chitinase inhibitor. The majority of the strains belonged to the genera Pseudomonas and Aeromonas and only 9% of the total of strains were Gram positive. From all the strains isolated, only 22 showed a wide growth range in all the pesticides tested and 4 of them were chosen for pesticide removal studies. The genetic identification of these strains showed their affiliation to Pseudomonas pseudoalcaligenes, Micrococcus luteus, Bacillus sp. and Exiguobacterium aurantiacum. These last two strains were those that showed the highest pesticide removal capacities and a high bacterial growth.

Growth of Azotobacter chroococcum in chemically defined media containing p-hydroxybenzoic acid and protocatechuic acid.

Growth and utilization of different phenolic acids present in olive mill wastewater (OMW) by Azotobacter chroococcum were studied in chemically defined media. Growth and utilization of phenolic acids were only detected when the microorganism was cultured on p-hydroxybenzoic acid at concentration from 0.01% to 0.5% (w/v) and protocatechuic acid at concentration from 0.01% to 0.3% (w/v) as sole carbon sources suggesting that only these phenolic compounds could be utilized as a carbon source by A. chroococcum. Moreover when culture media were added with a mixture of 0.3% of protocatechuic acid and 0.3% p-hydroxybenzoic acid, the microorganism degradated in first place protocatechuic acid and once the culture medium was depleted of this compound, the degradation of p-hydroxybenzoic acid commenced very fast.

Liberation of amino acids by heterotrophic nitrogen fixing bacteria.

Large amounts of amino acids are produced by nitrogen-fixing bacteria such as Azotobacter, Azospirillum, Rhizobium, Mesorhizobium and Sinorhizobium when growing in culture media amended with different carbon and nitrogen sources. This kind of bacteria live in close association with plant roots enhanced plant growth mainly as a result of their ability to fix nitrogen, improving shoot and root development suppression of pathogenic bacteria and fungi, and increase of available P concentration. Also, it has been strongly evidenced that production of biologically substances such as amino acids by these rhizobacteria are involved in many of the processes that explain plant-grown promotion. This paper reviews literature concerning amino acids production by nitrogen-fixing bacteria. The role of amino acids in microbial interactions in the rhizosphere and establishment of plant bacterial association is also discussed.

Production of amino acids by Azotobacter vinelandii and Azotobacter chroococcum with phenolic compounds as sole carbon source under diazotrophic and adiazotrophic conditions.

Azotobacter vinelandii strain ATCC 12837 and Azotobacter chroococcum strain H23 (CECT4435) were tested to grow in N-free or NH(4)Cl amended chemically defined media, with protocatechuic acid or sodium p-hydroxybenzoate as sole carbon (C) sources at a concentration of 2 mmol/L. Both substrates supported grow at similar rates than bacteria grown in control media amended with 2 mmol/L sodium succinate as C source. The two strains produced aspartic acid, serine, glutamic acid, glycine, hystidine, threonine, arginine, alanine, proline, cysteine, tyrosine, valine, methionine, lysine, isoleucine, leucine and phenylalanine after 72 h of growth in chemically defined media with 2 mmol/L of phenolic compounds or sodium succinate as sole C source amended or unamended with 0.1% (w/v) NH(4)Cl. Qualitative and quantitative production of all amino acids was not affected by the use of different C and N substrates.

Production of amino acids by Rhizobium, Mesorhizobium and Sinorhizobium strains in chemically defined media.

Five strains of Rhizobium spp, one strain of Mesorhizobium loti and two strains of Sinorhizobium meliloti were tested for their ability to grow in chemically-defined medium lacking growth factor. Qualitative and quantitative production of aspartic acid, serine, glutamic acid, glycine, histidine, threonine, arginine, alanine, proline, cysteine, tyrosine, valine, methionine, lysine, isoleucine, leucine, and phenylalanine was determined by the use of mannitol as sole carbon source. Strains of Rhizobium spp. and Sinorhizobium sp. produced all the amino acids analysed with the exception of cysteine and high biological levels of serine, glycine and alanine were detected after 2 days of culture in mineral medium. Strain U226 of M. loti only produced small amounts of amino acids and glutamic acid, histidine, arginine, cysteine, methionine, lysine and phenylalanine was not liberated into the media.

Effects of culture conditions on the production of polyhydroxyalkanoates by Azotobacter chroococcum H23 in media containing a high concentration of alpechín (wastewater from olive oil mills) as primary carbon source.

Large amounts of homopolymers containing beta-hydroxybutyrate (PHB) and copolymers containing beta-hydroxyvalerate (P[HB-co-HV]) are produced by Azotobacter chroococcum strain H23 when growing in culture media amended with alpechín (wastewater from olive oil mills) as the sole carbon source. Copolymer was formed when valerate (pentanoate) was added as a precursor to the alpechín medium, but it was not formed with the addition of propionate as a precursor. A. chroococcum formed homo- and copolymers of polyhydroxyalkanoates (PHAs) up to 80% of the cell dry weight, when grown on NH(4)(+)-medium supplemented with 60% (v/v) alpechín, after 48 h of incubation at 100 rev min(-1) and 30 degrees C. Production of PHAs by strain H23 using alpechín looks promising, as the use of a cheap substrate for the production of these materials is essential if bioplastics are to become competitive products.

Growth of moderately halophilic bacteria isolated from sea water using phenol as the sole carbon source.

Moderately halophilic bacteria utilizing phenol as the sole carbon source were isolated by selective enrichment from sea water. The isolate (Gram-negative motile rods) was identified as Deleya venusta. It grew well in the presence of up to 1600 mg/L of phenol and 8% NaCl under aerobic conditions. When the cells were treated with chloramphenicol prior to the addition of phenol they did not utilize added phenol, even after prolonged incubation. Thus, the enzymes necessary for phenol metabolism appeared to be inducible.

Production of B-group vitamins by two Azotobacter strains with phenolic compounds as sole carbon source under diazotrophic and adiazotrophic conditions.

Azotobacter vinelandii strain ATCC 12837 and A. chroococcum strain H23 (CECT 4435) were able to grow on N-free or NH4Cl-amended chemically-defined (Burk's) media, with protocatechuic acid (1-2 mmol 1(-1)) or sodium p-hydroxybenzoate (1-10 mmol 1(-1)) as sole carbon (C) sources. At a concentration of 2 mmol 1(-1), both substrates supported nitrogen fixation (acetylene reduction assay) at similar or higher rates than bacteria grown in control media amended with 2 mmol 1(-1) sodium succinate as C source. The two strains produced the B-group vitamins niacin, pantothenic acid, thiamine, riboflavin and biotin after 72 h of growth in chemically-defined media with 2 mmol 1(-1) protocatechuic acid, sodium phydroxybenzoate or sodium succinate as sole C source, either in N-free media or in media amended with 0.1% NH4Cl. Quantitative production of all vitamins was affected by the use of the different C and N substrates.

Effect of some herbicides on the production of lysine by Azotobacter chroococcum.

Production of lysine by Azotobacter chroococcum strain H23 was studied in chemically-defined media amended with different concentrations of alachlor, metolachlor, 2,4-D, 2,4,5-T and 2,3,6-TBA. The presence of 5, 10, and 50 micrograms/ml of alachlor or 2,3,6-TBA significantly decreased quantitative production of lysine. However, the presence 2,4-D or 2,4,5-T at concentrations of 10 and 50 micrograms/ml enhanced the production of lysine. Quantitative production of lysine was not affected as consequence of the addition of metolachlor to the culture medium, showing that the release lysine to the culture media by A. chroococcum was not affected by that herbicide.

Production of amino acids by free-living heterotrophic nitrogen-fixing bacteria.

Interest of microbial production of amino acids has been increased greatly since development of biotechnological methods. These methods represent a perspective way applied in a future large-scale manufacture of inexpensive amino acids. In this context, the isolation of producing organisms that may be exploited in the desing of alternative methods for the production of amino acids could be of primary importance.In this review we will describe the liberation of amino acids (methionine, lysine, arginine, tryptophane and glutamic acid) byAzotobacter andAzospirillum during growth in culture media with different carbon sources under diazotrophic and adiazotrophic conditions. These organisms may be useful in developing new methods for the industrial production of amino acids.

Effects of the herbicide alachlor on soil microbial activities.

: A study was made of the effects of one selected acetanilide herbicide, alachlor, at concentrations of 2.0-10.0 kg ha(-1) on bacterial populations, fungi, dinitrogen fixation bacteria, denitrifying bacteria, nitrifying bacteria, nitrogenase activity, acid and alkaline phosphatases, arylsulfatase and deshydrogenase. The presence of 2.0-10.0 kg ha(-1) of alachlor in the soil increased the total number of bacteria and fungi. The population of denitrifying bacteria increased significantly at concentrations of 5.0-10.0 kg ha(-1). However, aerobic dinitrogen fixing bacteria and nitrogenase activity decreased at alachlor concentrations of 3.5-10.0 kg ha(-1). Acid and alkaline phosphatases, arylsulfatase and dehydrogenase activity decreased significantly initially at concentrations of 5.0-10.0 kg ha(-1), but recovered to levels similar to those in the control. Nitrifying bacteria were not affected as a consequence of the addition of the herbicide to agricultural soil.

Toxicity of dicamba (2-methoxy-3,6-dichlorobenzoic acid) to Azotobacter vinelandii.

The effect of dicamba was studied in N-free medium inoculated with Azotobacter vinelandii ATCC 12837. Nitrogen fixation was determined by acetylene reduction. Dicamba at a concentration of 500 micrograms/mL had a strong inhibitory effect on nitrogenase activity. However, no inhibitory effect on microbial respiration was detected.