The use of a mixed culture of Geotrichum candidum,Candida krusei and Hansenula anomala for microbial protein production from whiskey distillery spent wash

Summary The use of a mixed culture of Geotrichum candidum, Hansenula anomala and Candida krusei for microbial protein production and treatment of whiskey distillery spent wash was investigated in both batch and continuous culture. Although capable of assimilating the same substrate constituents in pure culture the organisms used different constituents for growth in mixed continuous culture, allowing a stable mixed population to be established. The relative proportions of the three organisms in the population was dependent on the dilution rate.The mixed culture did not give superior yields or productivities, but did give proportionally greater COD reduction and would be expected to bemore stable and resistant to contamination.At a dilution rate of 0.10 h^–1 the mixed culture gave a biomass yield of 4.8 g l^–1, containing 55% crude protein, with a COD reduction of 31.5%, while in mixed batch culture a biomass yield of 12.5 g l^–1, containing 48% crude protein, was obtained with a COD reduction of 54.9%.     

Optimization of a biological process for treating potato chips industry wastewater using a mixed culture of Aspergillus foetidus and Aspergillus niger

Potato chips industry wastewater was collected and analyzed for biochemical oxygen demand (BOD), chemical oxygen demand (COD), total suspended solids (TSS) and total carbohydrates. Two Aspergillus species, A. foetidus and A. niger, were evaluated for their ability to grow and produce biomass and reduce the organic load of the wastewater. A. foetidus MTCC 508 and A. niger ITCC 2012 were able to reduce COD by about 60% and produce biomass 2.4 and 2.85 gl(-1), respectively. Co-inoculation of both Aspergillus strains resulted in increased fungal biomass production and higher COD reduction than in individual culture at different culture pH. pH 6 was optimum for biomass production and COD reduction. Amendment of the wastewater with different N and P sources, increased the biomass production and COD reduction substantially. Under standardized conditions of pH 6 and amendment of wastewater with 0.1% KH2PO4 and 0.1% (NH4)2 SO4, a mixed culture gave 90% reduction in COD within 60 h of incubation.     

Growth of Trichoderma harzianum and Myceliophthora thermophila in palm oil sludge

Uncentrifuged palm oil mill sludge (POS) diluted to about 50% (v/v in tap water) supports good mycelial growth of Myceliophthora thermophila and Trichoderma harzianum. Both of the selected fungi are non-toxic to mice. After 24 h M. thermophila grown in batch culture in POS yielded 28.6 g/l of mycelial biomass with biological oxygen demand (BOD) and chemical oxidation demand (COD) reductions of 72% and 74% respectively. T. harzianum yielded 24.4 g/l of mycelial biomass with BOD and COD reductions of 67% and 68% respectively. The crude protein of the mycelial biomass of M. thermophila and T. harzianum was twice that of untreated POS. T. harzianum showed amylolytic activity while M. thermophila was cellulolytic and lipolytic.     

Protozoan biomass relation to nutrient and chemical oxygen demand removal in activated sludge mixed liquor

The relationship between biomass concentration to nutrient and chemical oxygen demand (COD) removal in mixed liquor supplemented with sodium acetate was investigated, using three protozoan isolates and three different initial biomass concentrations (10(1), 10(2) and 10(3) cells/mL). The study was carried out in a shaking flask environment at a shaking speed of 100 rpm for 96 h at 25 degrees C. Aliquot samples were taken periodically for the determination of phosphate, nitrate, COD and dissolved oxygen, using standard methods. The results revealed remarkable phosphate removal of 82-95% at biomass concentration of 10(3)cells/mL. A high nitrate removal of over 87% was observed at all initial biomass concentration in mixed liquor. There was an observed COD increase of over 50% in mixed liquor in at the end of 96-h incubation and this was irrespective of initial biomass concentration used for inoculation. The study shows the trend in nutrient and COD removal at different biomass concentrations of the test isolates in mixed liquor.     

The mixed culture of microalgae Chlorella pyrenoidosa and yeast Yarrowia lipolytica for microbial biomass production

Microbial biomass which mostly generated from the microbial processes of bacteria, yeasts, and microalgae is an important resource. Recent concerns in microbial biomass production field, especially microbial lipid production for biofuel, have been focused towards the mixed culture of microalgae and yeast. To more comprehensive understanding of the mixed culture for microbial biomass, mono Chlorella pyrenoidosa, mono Yarrowia lipolytica and the mixed culture were investigated in the present work. Results showed that the mixed culture achieved significantly faster cell propagation of microalga and yeast, smaller individual cell size of yeast and higher relative chlorophyll content of microalga. The mixed culture facilitated the assimilation of carbon and nitrogen and drove the carbon flow to carbohydrate. Besides higher lipid yield (0.77 g/L), higher yields of carbohydrates (1.82 g/L), protein (1.99 g/L) and heating value (114.64 kJ/L) indicated the microbial biomass harvested from the mixed culture have more potential utilization in renewable energy, feedstuff, and chemical industry.

     

Yeast screening from avermectins wastewater and investigation on the ability of its fermentation

Wastewater from avermectin production is refractory with high output, high chemical oxygen demand (COD) concentration and high cost further. The current wastewater treatment technology, with little reuse of the organic material, needs high dilution ratio during treatment. Yeast single cell protein was produced from the wastewater of avermectins fermentation in this research. First, the yeast strain (H-1) which show enough tolerance to avermectins residue was screened from the wastewater, and it was identified as being most closely related to Candida tropicalis (100%, EF120592.1) using 18S rDNA gene sequence analysis. Second, its growth characteristics in the avermectins wastewater were studied. The dry biomass reached the maximal point of 10 g/L, the COD removal was up to 66.67%, and avermectins removal in the wastewater was 99.48% at the optimal condition that it was liquid volume 50 ml (250 ml flask), pH 4.0, temperature 30°C, inoculum volume 10% (V/V) and fermentation period 20 h. Third, the nutritional contents of dry yeast powder were determined, in which the water content was 8.12%, ash content was 5.18% and the crude protein was 40.02%. The dry yeast powder from avermectin waste liquor was promising to be used as a raw material or nitrogen source for commercial production of avermectins. The project was economically feasible by primary cost accounting.     

Influence of mixed culture conditions on yeast-wall hydrolytic activity of Bacillus circulans WL-12 and on extractability of astaxanthin from the yeast Phaffia rhodozyma

O kagbue , R.N. & L ewis , M.J. 1985. Influence of mixed culture conditions on yeast-wall hydrolytic activity of Bacillus circulans WL-12 and on extractability of astaxanthin from the yeast Phaffia rhodozyma. Journal of Applied Bacteriology 59 , 243–255.
In mixed culture Bacillus circulans WL-12 hydrolysed cell walls of Phaffia rhodozyma and rendered astaxanthin extractable from the yeast. pH control was critical to survival and lytic activity of the bacillus; the optimum range was 6.2–6.8. The optimum range of temperature was 20–24C. Glucose (1–2%) was efficient in minimizing catabolite repression of the lytic enzyme complex of the bacillus. Slow-feeding of glucose improved ultimate yields of lytic enzyme but did not acclerate yeast cell wall modification. A relatively high inoculum level of B. circulans accelerated modification of P. rhodozyma in the mixed culture: when the bacterial inoculum was four times that of the yeast, over 80% of total astaxanthin was extractable in 48 h. High bacterial inoculum size also stimulated yeast autolysis and necessitated early harvest of the mixed culture. Results obtained in shake flasks were duplicated in 5-litre fermentors and suggest that the mixed culture has potential industrial value for producing a biomass containing biologically-available astaxanthin. Extractability of astaxanthin was also achieved when mixed culture filtrate was incubated with pure cultured Phaffia cells. When suitably fortified with nutrients, the filtrate also supported simultaneous yeast growth and modification of the yeast cell walls. A scheme incorporating mixed culture with B. circulans WL-12 and re-use of culture filtrate has been proposed for enzymatic processing of Phaffia rhodozyma for inclusion in animal diets.     

Mixed culture of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris for lipid production from industrial wastes and its use as biodiesel feedstock

A mixed culture of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris was performed to enhance lipid production from industrial wastes. These included effluent from seafood processing plant and molasses from sugar cane plant. In the mixed culture, the yeast grew faster and the lipid production was higher than that in the pure cultures. This could be because microalga acted as an oxygen generator for yeast, while yeast provided CO(2) to microalga and both carried out the production of lipids. The optimal conditions for lipid production by the mixed culture were as follows: ratio of yeast to microalga at 1:1; initial pH at 5.0; molasses concentration at 1%; shaking speed at 200 rpm; and light intensity at 5.0 klux under 16:8 hours light and dark cycles. Under these conditions, the highest biomass of 4.63±0.15 g/L and lipid production of 2.88±0.16 g/L were obtained after five days of cultivation. In addition, the plant oil-like fatty acid composition of yeast and microalgal lipids suggested their high potential for use as biodiesel feedstock.     

粗甘油优化污泥脱水液培养小球藻过程的研究

污泥脱水液为污泥压缩过程产生的污水,因其含有N、P等营养物,可用于微藻的培养。但污泥脱水液碳氮比低,可利用碳源有限,影响微藻生长。本研究考察外加不同浓度(1 g/L,2 g/L,4 g/L,6 g/L)生物柴油副产物-粗甘油对污泥脱水液培养小球藻过程的影响。结果表明:1 g/L、2 g/L粗甘油浓度能促进小球藻生长,藻生物量为1.29 g/L、1.45 g/L;2 g/L粗甘油浓度下氨氮、总氮去除率达99.32%和97.52%。粗甘油被分解后易使培养体系pH降至7以下,使总磷去除率比对照组略低。外加1 g/L、2 g/L粗甘油组的COD去除量分别为553.00 mg/L和405.00 mg/L。藻细胞元素和傅里叶红外光谱分析表明补加粗甘油后藻细胞中C元素和H元素相对含量均明显增加,C元素含量约为对照组的1.5倍;2 g/L粗甘油组的蛋白质与脂类物质含量均高于对照组。MPBR半连续培养小球藻过程中,HRT为5 d时藻生物量维持在1.99~2.21 g/L,大约为批次生物量的1.50倍;氨氮、总氮、总磷、COD的去除率分别在96.26%~99.20%、92.44%~94.04%、53.63%~58.58%、59.44%~65.57%。     

Effectiveness of algae in the treatment of a wood-based pulp and paper industry wastewater

In this study, the ability of algae to treat a wood-based pulp and paper industry wastewater was investigated. Tests were performed in batch reactors seeded with a mixed culture of algae. Under different lighting and initial wastewater strength conditions, changes in COD, AOX and color contents of reactors were followed with time. Algae were found to remove up to 58% of COD, 84% of color and 80% of AOX from pulp and paper industry wastewaters. No remarkable differences were observed in COD and color when light intensity and wastewater strength were changed, while AOX removals were strongly affected. Algal species identification studies revealed that some green algae (Chlorella) and diatom species were dominant in the treatment. The study also showed that algae grew mixotrophically, while the main mechanism of color and organics removal from pulping effluents was partly metabolism and partly metabolic conversion of colored and chlorinated molecules to non-colored and non-chlorinated molecules. Adsorption onto algal biomass was not so effective.     

Industrial wastewater treatment in internal circulation bioreactor followed by wetlands containing emergent plants and algae

Wastewater treatment based on ecological principles is a low cost and highly desirable solution for the developing countries like Pakistan. The present study evaluated the effectiveness of biological treatment systems including Internal Circulation (IC) anaerobic bioreactor and constructed wetlands (CWs) containing macrophytes and mixed algal cultures for industrial wastewater treatment. The IC bioreactor reduced COD (52%), turbidity (89%), EC (24%) of the industrial wastewater. However, the effluents of IC bioreactor did not comply with National Environmental Quality Standards (NEQS) of Pakistan. Post-treatment of IC bioreactor effluents was accomplished in CW containing macrophytes (Arundo donax and Eichhornia crassipes) and mixed algal culture. The CWs planted with macrophytes lowered the concentrations of COD (89%) and turbidity (99%). CWs with algal biomass were not effective in further polishing the effluent. Inhibition of algal biomass growth was observed due to physicochemical characteristics of wastewater. The integrated treatment system consisting of IC bioreactor and macrophytes was found more suitable option for industrial wastewater treatment.     

Growth and production of biomass of Rhodovulum sulfidophilum in sardine processing wastewater

AIMS: Rhodovulum sulfidophilum was grown in sardine processing wastewater to assess growth characteristics for the production of bacterial biomass with simultaneous reduction of chemical oxygen demand. METHODS AND RESULTS: Growth characteristics were compared in diluted and undiluted, settled and non-settled wastewater growing in anaerobic light and aerobic dark conditions; and also at different agitation speeds. The highest biomass (8.75 g l(-1)) and a reduction in chemical oxygen demand of 71% were obtained in unsettled, undiluted wastewater after 120 h culture with 15% inoculum. In settled wastewater, highest biomass (7.64 g l(-1)) and a COD reduction of 77% was also obtained after 120 h. Total biomass was higher (4.34 g l(-1)) after 120 h culture in anaerobic light compared to (3.23 g l(-1)) in aerobic dark growth. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Better performance, mean of total biomass (6.97 g l(-1) after 96 h), total carotenoids (4.24 mg g(-1) dry cell from 24 h) and soluble protein (431 microg ml(-1) after 96 h) were obtained from aerobic dark culture at 300 rev min(-1). The COD reduction, however, was lower (69%) after 96 h culture. Thus, the benefits in the production of bacterial biomass in non-sterilized sardine processing wastewater with the reduction of chemical oxygen demand could be achieved.     

Biodegradation of lignocellulosic waste by Aspergillus terreus

Biodegradation of lignocellulosic waste by Aspergillus terreus is reported for the first time. This isolate produced 250 CMCase (carboxymethyl cellulase or endoglucanase) U.ml(-1) and biodegraded hay and straw during 3 days and the biomass production on straw was 5g.L(-1) dry weight from 0.25 cm2 inoculated mycellium. This strain secreted endocellulases and exocellulases in the culture medium, but some of the enzymes produced, remained cell membrane bound. Cell bound enzymes were released by various treatments. The highest amount of endoglucanase and exoglucanase was released when the cells were treated with sonication. Aspergillus terreus was added to two tanks containing sugar wastewater and pulp manufacturing waste, as a seed for COD removal. This fungus reduced the COD by 40-80 percent, also, ammonia was reduced from 14.5 mM to 5.6 mM in sugar beet wastewater. The effects of crude enzyme of this fungus for COD removal was studied.     

Rhodovulum sulfidophilum in the treatment and utilization of sardine processing wastewater

AIMS: Rhodovulum sulfidophilum was grown in settled undiluted and nonsterilized sardine processing wastewater (SPW). The aims were to evaluate the effects of inoculum size and media on the biomass production with simultaneous reduction of chemical oxygen demand (COD). METHODS AND RESULTS: Three levels of inoculum size (10, 20 and 30% v/v) developed in glutamate-malate media (GMM) or settled and undiluted SPW were compared. The highest biomass (4.8 g l-1) was obtained after 96-h culture with 20% (v/v) inoculum size, but the reduction in COD of SPW was the highest (85%) after 120-h culture with a 30% (v/v) inoculum developed in GMM. In cultures with inoculum developed in SPW the COD reduction in SPW was 79-83%. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Inoculum developed in GMM supported good growth of Rv. sulfidophilum in settled undiluted SPW and subsequent reduction in COD. A conceptual model was proposed for the treatment and utilization of SPW.     

Biomass production of yeast isolate from salad oil manufacturing wastewater

Conversion of oil-rich salad oil manufacturing wastewater (SOMW) into protein source for animal feed through biomass production of yeast isolate was investigated in this study. Five species of yeasts, including Rhodotorula rubra, Candida tropicalis, C. utilis, C. boidinii, Trichosporon cutaneum, were isolated from SOMW following enrichment culture. Of them, C. utilis was chosen as the sole biomass producer in the study due to its greatest oil uptake rate, 0.96 kg oil kg(-1) biomass d(-1), and highest specific growth rate, 0.25 h(-1). The cells of C. utilis contained 26% protein, 9% crude lipid, 55% carbohydrate and balanced amino acid compositions. The initial N:C ratio in SOMW drastically influenced oil reduction efficiency, biomass production and protein content of C. utilis, and therefore a range between 1:6 and 1:8 was recommended in consideration of these three factors simultaneously.     

Responses of pea and wheat to textile wastewater reclaimed by suspended sequencing batch bioreactors

Availability of good quality water for crop irrigation is a big challenge in developing countries due to limited resources of clean water. Textile industry consumes a huge amount of water during dyeing process and consequently it releases high strength wastewater into wastewater streams. The present study was designed with the objective to use textile wastewater treated in sequencing batch bioreactor for irrigation purpose. Wastewater containing 100 mg/L reactive black-5 azo dye amended with different co-substrates was treated using mixed liquor suspended solids (MLSS) and two previously isolated dye-degrading bacterial strains (Psychrobacter alimentarius KS23 and Staphylococcus equorum KS26). About 90% color and COD removal in case of dye-containing wastewater amended either with mineral salts + yeast extract or only yeast extract was achieved in 24 h after treatment with mixed culture (MLSS + KS23 + KS26). The treated wastewater was applied for irrigation of pea and wheat plants under controlled conditions. Untreated dye-contaminated wastewater was used as a control for comparison. A significant positive effect of treated dye wastewater amended with different co-substrates on the seed germination index, root and shoot length and biomass was observed in response to application of dye-containing wastewater treated with MLSS and dye-degrading bacterial strains compared to untreated control. Results of this study reveal that the dye-degrading microbial cultures could be used to enhance the treatment efficiency of dye-contaminated wastewater that can be utilized for irrigation of crops and biomass production.     

A model for continuous fermentations with amylolytic yeasts

A two-stage, associative fermentation process is more effective for continuous yeast biomass production from starch than a single-stage mixed culture fermentation process. By operating two stages, competition for the same growth limiting substrate is reduced leading to efficient starch utilization. In this article, a mathematical model has been proposed for continuous, two-stage fermentation with a pure culture, amylolytic yeast in the first stage and a mixed culture second stage with a faster growing, nonamylolytic yeast. The model parameters were determined for Saccharomycopsis fibuligera and Candida utilis in continuous, single-stage, pure cultures. In the two-stage model, the effects of changes in dilution rate on biomass, amylase, reducing sugar, and starch concentration, and ratio of stage volumes on microbial composition are discussed and compared with experimental data.     

Biodegradable polymers from organic acids by using activated sludge enriched by aerobic periodic feeding

This article describes a new process for the production of biopolymers (polyhydroxyalkanoates, PHAs) based on the aerobic enrichment of activated sludge to obtain mixed cultures able to store PHAs at high rates and yields. Enrichment was obtained through the selective pressure established by feeding the carbon source in a periodic mode (feast and famine regime) in a sequencing batch reactor. A concentrated mixture of acetic, lactic, and propionic acids (overall concentration of 8.5 gCOD L(-1)) was fed every 2 h at 1 day(-1) overall dilution rate. Even at such high organic load (8.5 gCOD L(-1) day(-1)), the selective pressure due to periodic feeding was effective in obtaining a biomass with a storage ability much higher than activated sludges. The immediate biomass response to substrate excess (as determined thorough short-term batch tests) was characterized by a storage rate and yield of 649 mgPHA (as COD) g biomass (as COD)(-1) h(-1) and 0.45 mgPHA (as COD) mg removed substrates (as COD(-1)), respectively. When the substrate excess was present for more than 2 h (long-term batch tests), the storage rate and yield decreased, whereas growth rate and yield significantly increased due to biomass adaptation. A maximum polymer fraction in the biomass was therefore obtained at about 50% (on COD basis). As for the PHA composition, the copolymer poly(beta-hydroxybutyrate/beta-hydroxyvalerate) with 31% of hydroxyvalerate monomer was produced from the substrate mixture. Comparison of the tests with individual and mixed substrates seemed to indicate that, on removing the substrate mixture for copolymer production, propionic acid was fully utilized to produce propionylCoA, whereas the acetylCoA was fully provided by acetic and lactic acid.     

Detoxification of olive mill wastewaters by Moroccan yeast isolates

A total of 105 yeast strains were isolated from Moroccan olive oil production plants and evaluated for their ability to grow in olive oil mill wastewaters (OMW). The 9 isolates that grew best on OMW were selected for further study to evaluate their effect on removal of organic pollutants and OMW phytotoxicity (barley seed germination test). The results showed that at least four yeast isolates effectively lowered the toxicity of this effluent in addition to providing very useful materials in terms of both yeast biomass (6 g/l DW) and an irrigation fluid. This group of yeast isolates significantly reduced the concentration of total phenols (44% removal) and Chemical Oxygen Demand, COD (63% removal). The best germination rate of 80% for undiluted OMW was obtained for strain Candida holstii that also increased the pH from 4.76 to 6.75. Principal component analysis of the results obtained for the best yeast strains confirmed the importance of COD and total phenol reduction along with increase of organic nitrogen and final pH for the improvement of germination rates and phytotoxic reduction. This study has highlighted the potential of indigenous yeasts in detoxification of olive mill wastewaters.     

Performance of the anaerobic baffled reactor under steady-state and shock loading conditions

The anaerobic baffled reactor (ABR) contains a granulated, mixed anaerobic culture segregated into compartments. Operation of four reactors under a range of hydraulic retention times showed that this novel reactor design offers highly efficient performance in the conversion of carbon in the feed stream to methane and carbon dioxide. The design parameter varied was the number of compartments. COD removal at 20 h retention time was routinely over 95% in all reactors, with low washout of biomass. Very high specific reaction rates were achievable (although with a loss of efficiency) at low biomass concentrations and high loading rates. In order to optimize volumetric reaction rates, a tradeoff has to be made between high biomass concentration, granule size, and the resulting mass transfer limitations. Formate is shown to be an important intermediate in the process under conditions of high loading.