The effects of algal inoculation on chemical oxygen demand (COD) and total nitrogen (TN) removal, and indigenous bacterial dynamics were investigated in municipal wastewater. Experiments were conducted with municipal wastewater inoculated with either Chlorella vulgaris AG10032, Selenastrum gracile UTEX 325, or Scenedesmus quadricauda AG 10308. C. vulgaris and S. gracile as fast growing algae in municipal wastewater, performed high COD and TN removal in contrast to Sc. quadricauda. The indigenous bacterial dynamics revealed by 16S rRNA gene amplification showed different bacterial shifts in response to different algal inoculations. The dominant bacterial genera of either algal case were characterized as heterotrophic nitrifying bacteria. Our results suggest that selection of indigenous bacteria that symbiotically interact with algal species is important for better performance of wastewater treatment.
Effects of phosphorous (P) and methylglyoxal (MG) on the cell number, dry weight, chlorophyll content, photosynthetic and respiratory rate, phosphate uptake and protein content of green algae (Scenedesrnus obliquus) were studied. The algal cell number from the medium treated with 0.5-1.0 mM of MG at 1/2 P or 1/4 P concentration was significantly lower than those of algae treated :with full strength of phosphrous in medium. The inhibitory effect of MG on algal cell division was enhenced at low concentration of phosphorous in medium. At the beginning of logrithmic phase of algal growth, the mean dry weight of algae from the medium without MG-treatment in 1/2 P media was significantly higher than that of algae treated with MG. After logrithmic phase of growth cycle, the mean dry weight of algae from the medium with 1.0 mM of MG-treatment in 1/4 P media was significantly lower than that of algae treated with or without MG. At logrithmic phase of algal growth, there were significant differences in the chlorophyll content among all groups of tested algae with various concentrations of P and MG. At 15 days after inoculation, the mean chlorophyll content per algal cell from the media without MG-treatment in 1/2P was significantly higher than that of other cells from MG-treated media. The adverse effect of MG at concentration of 0.5-1.0mM in 1/2 and 1/4 P media on photosynthetic rate was observed. The mean photosynthetic rate of algal cell without P and MG treatment at 15 days after inoculation was significantly higher than that of MGtreated algae. After logarithmic phase, the algal cell treated with 0.5mM of MG with full strength of phosphorous showed significantly high respiratory rate than that of other cell groups. There were significant differences in mean phosphate uptake rate among all groups of Scenedesmus obliquus at logarithmic phase. At 12 days after inoculation, phosphate uptake rate per each algal cell from the basic media without MG and P treatment was rapidly reduced which shows early introduction to stationary phase.
Although chemical oxygen demand (COD) is an important issue for wastewater treatment, COD reduction with microalgae has been less studied compared to nitrogen or phosphorus removal. COD removal is not efficient in conventional wastewater treatment using microalgae, because the algae release organic compounds, thereby finally increasing the COD level. This study focused on enhancing COD removal and meeting the effluent standard for discharge by optimizing sludge inoculation timing, which was an important factor in forming a desirable algae/bacteria consortium for more efficient COD removal and higher biomass productivity. Activated sludge has been added to reduce COD in many studies, but its inoculation was done at the start of cultivation. However, when the sludge was added after 3 days of cultivation, at which point the COD concentration started to increase again, the algal growth and biomass productivity were higher than those of the initial sludge inoculation and control (without sludge). Algal and bacterial cell numbers measured by qPCR were also higher with sludge inoculation at 3 days later. In a semi-continuous cultivation system, a hydraulic retention time of 5 days with sludge inoculation resulted in the highest biomass productivity and N/P removal. This study achieved a further improved COD removal than the conventional microalgal wastewater treatment, by introducing bacteria in activated sludge at optimized timing.
The growth of Scenedesmus quadricauda (Trup.) Breb. is enhanced by methylyoxal (MG), a general inhibitor of cell division, at threshold concentration in conjunction with reatment timing relative to growth stage. The stimulatory effect of MG on algal cell growth was most significant with 2.27-fold of untreated algal culture in cell number when 0.5 mM of MG was added to the algal culture at the beginning of logarithmic phase with an initial MG concentration of 0.535 mg $MG/10^6cell$. A Specific growth rates (SGRs) of MG-treated cultures were rapidly increased at the beginning of logarithmic phase with 1.89-fold of untreated algal culture. Cultures inoculated with high cell numbers of 2.4 to 4.8 X $10^4$ cells/ml were less sensitive to 0.5 mM of MG treatment. The algal cell division was ranged from 0.392 to 0.924 mg MG/106 cell. If the cell number of an algal culture at the time of inoculation was low (0.6 X $10^4$ cells/ml) and MG was added before logarithmic phase, the cell number of 0.5 mM of MG-treated cultures were lower than those of controls. In algal cultures treated with high concentrations of MG (1.0 mM and 2.0 mM), the algal growth was inhibited. Photosynthetic rate of growth-enhanced algal by 0.5 mM of MG was significantly higher than that of untreated or 1.0 mM of MG-treated algal cell, while there was no significant difference among those groups in respiratory rate. Pyruvate concentration in 0.5 mM of MG-treated culture was incrcased agter methylglyoxal trcatment.
Park, Eun-Ju;Kim, Gwang-Hoon;Cho, Yong-Chul;Cho, Eun-Seob
Journal of the korean society of oceanography
/
v.34
no.4
/
pp.214-219
/
1999
Cells of the dinoflagellate Gyrodinium impudicum form characteristic chains, which are associated with sugar accumulated on the cell surface. To resolve the relationship between chain-formation and cell surface sugar accumulation, confocal microscopy was used to observe sugar accumulation points in the vegetative cells and long chain-forming cells of G. impudicum cells treated with fluorescent-tagged ConA. In the stationary and exponential phases, both vegetative cells and chain-forming cells were similar to each other in fluorescent intensity. There was no evidence that chain-forming cells had a specific location for sugar accumulation on the cell surface. Most of the cells formed 2-cell chains one day after inoculation, but longer chains consisting of 4-8 cells increased markedly in 4day and 8 day cultures. Gyrodinium impudicum chains usually consist of more than four cells, and chains of 8 or even 16 cells can be observed in mature cultures. Temperature played an importantrole in chain-formation of the cells, threshold temperature for the development of long chain-formation was 19 $^{\circ}$C.
Lee, Soon Jeong;Jee, Bo Young;Son, Maeng-Hyun;Lee, Sang-Rae
ALGAE
/
v.32
no.2
/
pp.155-160
/
2017
Red rot disease has caused a major decline in Pyropia (Nori) crop production in Korea, Japan, and China. To date, only Pythium porphyrae (Pythiales, Oomycetes) has been reported as the pathogen causing red rot disease in Pyropia yezoensis (Rhodophyta, Bangiales). Recently, Pythium chondricola was isolated from the infected blades of Py. yezoensis during molecular analyses using the mitochondrial cox1 region. In this study, we evaluated the pathogenicity of P. chondricola as an algal pathogen of Py. yezoensis. Moreover, a new cox2 marker was developed with high specificity for Pythium species. Subsequent to re-inoculation, P. chondricola successfully infected Py. yezoensis blades, with the infected regions containing symptoms of red rot disease. A novel cox2 marker successfully isolated the cox2 region of Pythium species from the infected blades of Py. yezoensis collected from Pyropia aquaculture farms. cox2 sequences showed 100% identity with that of P. chondricola (KJ595354) and 98% similarity with that of P. porphyrae (KJ595377). The results of the pathogenicity test and molecular analysis confirm that P. chondricola is a new algal pathogen causing red rot disease in Pyropia species. Moreover, it could also suggest the presence of cryptic biodiversity among Korean Pythium species.
Journal of Korean Society of Environmental Engineers
/
v.32
no.4
/
pp.333-340
/
2010
Wastewater treatment using algal communities and biodiesel production from wastewater-cultivated algal biomass is a promising green growth technology. In literature, there are many studies providing information on algal species producing high content of lipid. However, very little is known about adaptability and wastewater treatability of such high-lipid algal species. In this study, we attempted to characterize algal biomass production and wastewater treatability of high-lipid algal species under municipal wastewater condition. For this, four known high-lipid algal strains including Chlorella vulgaris AG 10032, Ankistrodesmus gracilis SAG 278-2, Scenedesmus quadricauda, and Botryococcus braunii UTEX 572 were individually inoculated into municipal wastewater where its indigenuous algal populations were removed prior to the inoculation, and the algae-inoculated wastewater was incubated in the presence of light source (80${\mu}E$) for 9 days in laboratory batch reactors. During the incubations, algal biomass production (dry weight) and the removals of dissolved organics (COD), nitrogen and phosphorous were measured in laboratory batch reactors. According to algal growth results, C. vulgaris, A. gracilis and S. quadricauda exhibited faster growth than indigenuous wastewater algal populations while B. braunii did not. The wastewater-growing strains exhibited efficient removals of total-N, ${NH_4}^+$-N, Total-P and ${PO_4}^{3-}$-P which satisfy the Korea water quality standards for effluent from municipal wastewater treatment plants. A. gracilis and S. quadricauda exhibited efficient and stable treatability of COD but C. vulgaris showed unstable treatability. Taken together with the results, A. gracilis and S. quadricauda were found to be suitable species for biomass production and wastewater treatment under municipal wastewater condition.
Park , Jong-Gyu;Hur, Hyun-Jung;Coats, D.Wayne;Yih, Won-Ho
ALGAE
/
v.22
no.4
/
pp.287-295
/
2007
Infection of free-living dinoflagellates by endoparasitic dinoflagellates of the genus Amoebophrya are thought to have significant impacts on host population dynamics and have long been proposed to be a potential biological agent for controlling harmful algal bloom (HAB). To understand the impact of Amoebophrya on particular host species, however, it is necessary to quantify aspects the parasites life cycle. Here we used cultures of Amoebophryahost systems from Jinhae Bay, Korea to determine, parasite generation time, and dinospore survival and infectivity. The proportion of host cells infected by Amoebophrya sp. changed sharply from 5% to 87% with increasing dinospore:host inoculation ratios. In the absence of H. triquetra, most free-living dinospores died within 72 hours and their ability to infect host cells decreased remarkably in a day. The relatively short free-living phase of Amoebophrya suggests that the spread of infections is most likely to occur during seasons of high host abundance, as that is when dinospores have the greatest chance of encountering host cells. Infection of host cells inoculated with dinospores during the day was higher than when inoculated during the night, suggesting that infection rates might be related to environmental light conditions and/or diurnal biological rhythm of host species. Total generation times of parasite strains from a thecate dinoflagellate Heterocapsa triquetra were nearly the same regardless of dinospore:host inoculation ratios, representing 54 ± 0.5 h in a 1:1 ratio and 55 ± 1.2 h in a 20:1 ratio. Dinospore production of Amoebophrya sp. infecting Heterocapsa triquetra was estimated to be 125 dinospores per a strain of Amoebophrya sp. There is a growing need to maintain a variety of host-parasite systems in culture and to examine their autecology under various environmental conditions. Such studies would be very helpful in understanding ecological role of these parasites, their overlooked importance in the flow of material and energy in marine ecosystem, and their practical use as biological control agents applied directly to areas affected by HAB.
In the process of the incorporation of orthophsphate into protein and other cell constituents, the role of inorganic polyphosphate and RNA-polyphosphate complex and the correlation between them were pursued by analyzing the contents of $^{32}P$ and total P in various fractions of Chlorella cells, which had been uniformly labeled with $^{32}P$ before the inoculation in a normal "cold" medium or P-free medium during the culture. The effects of ionizing radiation and various micronutritional-element deficiencies on the phosphate incorporation into, and biosynthesis of, protein and other introgenus compounds in the cells were also observed. When the uniformly $^{32}P$-labeled algae were grown in a normal "cold" medium the contents of $^{32}$ P in the fractions of protein, DNA and RNA-polyphosphate complex increased, but those in the fraction of acid-insoluble polyphosphate decreased. On the other hand, amount of $^{32}P$in the fraction of RNA was almost unchanged in spite of rapid increase of the total P. In the growing period of $^{32}P$-labeled algae in a P-free medium, amounts of $^{32}P$ in the fractions of DNA, protein and lipid increased, while those in the fractions of RNA-polyphosphate and inorganic polyphosphates decreased. When the algal cells were irradiated with about 70, 000r of gamma-rays before the inoculation in the medium, amounts of phosphate in the fractions of DNA, RNA, nucleotides and protein decreased during the culture, compared with those of the control. However, the phosphate content in the fraction of acid-insoluble polyphosphate of the irradiated cells increased than those of the control. In the growing period of the algae in a Mo-free, medium, amounts of acid-soluble total phosphate and nucleotides of the cells increased, while the amounts of residual protein and RNA decresed compared with those of the normal cells. Amounts of alkali-labile protein and phospholipid of the cells grown in a B-free medium decreased, whereas amount of phosphate in acid-soluble fraction increased compared with the control. In general, the contents of protein and RNA in each microelement deficient cells decreased more or less, compared with those in the normal cells.in the normal cells.
To isolate alga-lytic bacteria, a number of samples were collected from Lake of Sukchon and Pal'tang reservoir where cyanobacteria blooming occurred. HY0210-AK1, which exhibited high alga-lytic activity, was isolated using Anabaena cylindrica lawn. The morphological and biochemical characteristics of the isolate HY0210-AK1 were very similar to that of the genus Rhizobium. Taxonomic identification including 16S rDNA base sequencing and phylogenetic analysis indicated that the isolate Hy0210-AK1 had a 99.1% homology in its 16S rDNA babe sequence with Sphingobium herbicidovorans. A. cylindrica NIES-19 was susceptible to the alga-lytic bacterial attack. The growth-inhibiting offset of the bacterium was not different on A. cylindrica NIES-19 when Sphingobium herbicidovorans HY0210-AK1 was in the lag, exponential, and stationary growth phase, although the alga-Iytic effect of S. herbici-dovorans HY0210-AK1 that in stationary growth phase was somewhat pronounced at the first time of inoculation. When S. herbicidovorans HY0210-AK1 was inoculated was inoculated with $1\times 10^{8}$ CFU $ml^{-1}$ together with A cylindrica NIES-19, the bacterium proliferated and caused algal lysis. A. cylindrica NIES-19 died when S. herbicidovorans HY0210 AKl was added to the algal culture but not when duly the filtrates from the bacterial culture was added. This suggests that extracellular substances are not responsible for inhibition of A. cylindrica NIES-19 and that algal Iysis largely attributed to direct interaction between S. herbicidovorans HY0210-AK1 and A. cylindrica NIES-19. The alga-lytic bacterium HY0210-AK1 caused cell lysis and death of three strain of Micro-cystis aeruginosa, but revealed no alga-Iytic effects on the Stephanodiscus hantzschii.
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