Effect of trace metals on halophilic and mixed cultures in anaerobic treatment

Trace metals are known to stimulate the activity of anaerobic methanogenic bacteria involved in the degradation of organic matter. The objective of this research was to investigate the effect of three trace metals, iron, nickel and cobalt on the activity of halophilic methanogens, digester sludge and a mixed culture of halophilic bacteria and digester sludge, at various salt concentrations. Iron slightly stimulated both initial and maximum specific methanogenic activity (SMA) of halophilic methanogens at all salt concentrations. Iron addition was also able to increase both initial and maximum SMA of the mixed culture. Iron could not stimulate either initial or maximum SMA of digester sludge at salt concentrations of more than 10 g/L. Adding cobalt showed no benefit on the initial and maximum SMA of halophilic methanogens. A slight increase in the initial SMA was observed in mixed culture when cobalt was injected to the system containing sodium concentrations of 10–30 g/L. At salt concentrations above 40 g/L, cobalt could stimulate the maximum SMA of mixed culture. No benefits were found when cobalt was added to the digester sludge at sodium concentrations higher than 20 g/L. Nickel was observed to increase the initial SMA of digester sludge at all salt concentrations, and the maximum SMA of mixed culture at salt concentrations of 30–50 g/L.     

Anaerobic digester sludge as nutrient source for culturing of microalgae for economic biodiesel production

After decades of ‘living dangerously’, the human kind has paused to think of Mother Nature. Alternate energy sources are being developed as a part of this realization. The use of indigenous sources of nutrients would considerably bring down the cost of production. A mixed consortium of Chlorella vulgaris, Arthrospira platensis, and tap water-originated Scenedesmus dimorphus was cultured using natural seawater, anaerobic digester sludge, and its growth was compared with synthetic commercial media like Zarrouk’s media, NaNO₃ media, and NH₄Cl media. A spectrophotometric method was standardized for regular biomass estimation. The dry biomass of a 15-day pure, batch culture of Arthrospira was found to yield around 600 mg/L in anaerobic digester sludge, whereas the pure batch culture of chlorella’s growth was hindered mainly due to the presence of bacteria and grazers. Regular microscopic observation and biomass monitoring revealed a drastic reduction in grazing activity, with the use of autoclaved AD sludge, resulting in a strong and stable microalgal mixed consortia. The consortia growth in AD sludge was found to be better than with the synthetic media with no cost of nutrient. The mixed consortia yielded a biomass up to 600 mg/L and lipid of 21.18%. The lipid generated from AD sludge had around 95% unsaturates and contained around 5% omega-3 fatty acids. The use of anaerobic digester sludge in a non-sterile condition reduces the total cost of the biodiesel production process as a whole and introduces a decentralized system for waste water treatment as well.     

Start-up of halophilic nitrogen removal via nitrite from hypersaline wastewater by estuarine sediments in sequencing batch reactor

Nitrogen removal from hypersaline wastewater was successfully started up by inoculating estuarine sediments for 140 days. Efficient ammonia and total nitrogen removal was sustained under specific ammonia loading of 0.016–0.139 kg N/[kg VSS day] in a sequencing batch reactor. Stable nitrite accumulation was observed during nitrification. The specific ammonia consumption rate was higher than the value of freshwater activated sludge and salt-acclimated freshwater activated sludge. With methanol as carbon source, specific nitrite reduction rate of halophilic denitrifiers was much less than the freshwater counterpart. Halophilic activated sludge was characterized as good settling and flocculation prosperity with small floc size and net-like sludge structure. The abundance of ammonia-oxidizing bacteria outnumbered ammonia-oxidizing archaeas in both estuarine sediments and the activated sludge. Nitrifier population was dominated by the halophilic members of genus Nitrosomonas. This study demonstrated the application of mixed halophilic consortia for efficient nitrogen removal, overcoming the limits and difficulties of applying freshwater bacteria for saline wastewater treatment.     

Effect of sodium chloride on the metabolic activity of halophilic bacteria isolated from the Lake Gardno estuary

The effect of different concentrations of sodium chloride on the metabolic activity of halophilic bacteria derived from the Lake Gardno estuary was studied using respirometric methods. The influence of salt was examined in two ways: the bacteria were preincubated in various concentrations of sodium chloride, and the cells used for respirometric measurements were supplied with substrates dissolved in water amended with various concentrations of the salt. Generally, an increase in the concentration of sodium chloride in the medium used for preincubation resulted in an increase in metabolic activity. Conversely, a decrease was noted when solutions of increasing salt content were employed to dissolve the substrate. The decrease in oxygen uptake noted when increased amounts of salt were added is proposed to be the result of the short period allowed for adaptation of cells to the higher salt concentrations.     

Study of syntrophic anaerobic digestion of volatile fatty acids using enriched cultures at mesophilic conditions

Volatile fatty acids are the most important intermediates in anaerobic digestion, and their degradations are extremely complicated thermodynamically. In this research, syntrophic anaerobic digestion of volatile fatty acids using enriched acetogenic and methanogenic cultures in a batch reactor at mesophilic conditions was investigated. Interactive effects of key microbiological and operating variables (propionic, butyric and acetic acids, retention time and methanogen to acetogen populations ratio) on the anaerobic degradation of volatile fatty acids were analyzed. Acetogenic and methanogenic anaerobes in the granular sludge from an up-flow anaerobic sludge blanket reactor were enriched at mesophilic conditions within a period of four weeks, separately. Enriched cultures were mixed with known proportions and then used in the bioreactor. Experiments were carried out based on central composite design and analyzed using response surface methodology. Four parameters (final concentrations of propionic, butyric and acetic acids and biogas production) were directly measured as response. Also, the optimum conditions for volatile fatty acid degradation were found to be 937.5 mg/L, 3275.5 mg/L, 2319.5 mg/L, 45 h and 2.2 proportions for propionic acid, butyric acid, acetic acid, retention time and methanogen to acetogen populations ratio, respectively (corresponding to maximum volatile fatty acid removal efficiencies and biogas production). The results of the verification experiment and the predicted values from the fitted correlations at the optimum conditions were in close agreement at a 95% confidence interval. The present study provides valuable information about the interrelations of quality and process parameters at different values of microbiological and operating variables.     

Preservation of thermophilic mixed culture for anaerobic palm oil mill effluent treatment by convective drying methods

The generation of huge amount of liquid waste known as palm oil mill effluent is a major problem in oil palm industry. Meanwhile, anaerobic biodegradation of such organic effluent at thermophilic condition is a promising treatment technology due to its high efficiency. However, storage and transportation of thermophilic mixed culture sludge are challenging due to constant biogas generation and heating requirement. Hence, drying of thermophilic sludge was conducted to obtain dormant thermophiles and thus enables easier handling. In this study, thermophilic sludge was dried using heat pump at 22 and 32 °C as well as hot air oven at 40, 50, 60, and 70 °C. Subsequently, quality of dried sludge was examined based on most probable number enumeration, chemical oxygen demand, and methane yield. Average drying rate was found to increase from 3.21 to 17.84 g H₂O/m² min as drying temperatures increases while average moisture diffusivity values ranges from 5.07 × 10^?9 to 4.34 × 10^?8 m²/s. Oven drying of thermophilic mixed culture resulted in highest chemical oxygen demand removal and lowest log reduction of anaerobes at 53.41% and 2.16, respectively, while heat pump drying resulted in the highest methane yield and lowest log reduction of methanogens at 53.4 ml CH₄/g COD and 2.09, respectively. To conclude, heat pump at 22 °C was most suitable drying technique for thermophilic mixed culture as the original methane-producing capability was largely retained after drying, at a slightly lower yet still comparable chemical oxygen demand removal when palm oil mill effluent was treated with the rehydrated culture.     

Diphytanyl glycerol ether distributions in sediments of the Orca Basin

Archaebacterially produced diphytanyl glycerol ether (DPGE) was examined in core sediments from the Orca Basin, an anoxic hypersaline basin in the northwestern Gulf of Mexico, to observe its spatial variability and potential origin. A differential extraction protocol was employed to quantify the isopranyl glycerol ethers associated with unbound, intermediate-bound, and kerogen-bound lipid fractions. Archaebacterial lipids were evident at all depths for the unbound and intermediate-bound fractions. Concentrations of DPGE ranged from 0.51 to 2.91 micrograms/g dry sediment at the surface and showed secondary maxima deeper in basin sediments. Intermediate-bound DPGE concentrations exhibited an inverse relationship to unbound DPGE concentrations. Kerogen-bound DPGE concentrations were normally below detection limits. Earlier studies describing the general homogeneity of lipid components within the overlying brine and at the brine/seawater interface suggest that the large-scale sedimentary DPGE variations observed in this study result from spatial and temporal variations in in situ production by methanogenic or extremely halophilic archaebacteria.     

Effect of different growth parameters on endoglucanase enzyme activity by bacteria isolated from coir retting effluents of estuarine environment

The cellulolytic enzyme-endoglucanase activity against coir fibre, a major biowaste by bacteria such as Cellulomonas, Bacillus and Micrococcus spp. isolated from coir retting effluents of estuarine environment was studied. The enzyme assay was carried out by using various concentrations (0.5 ? 2%) of substrate of coir powder as a carbohydrate in different pH (5–9) and temperature (20–50 °C). The enzyme activity was minimum in 0.5% substrate concentration at lower pH 5 (0.0087, 0.0143 and 0.0071 U/mL) and at 20 °C temperature (0.0151, 0.0154 and 0.0122 U/mL) by the bacterial strains such as Cellulomonas, Bacillus and Micrococcus spp respectively. Then this level was increased and reached maximum at the neutral pH (0.0172, 0.0165 and 0.0121 U/mL) and at 40 °C (0.0336, 0.0196 and 0.0152 U/mL) by the selected bacterial species. Further increase of pH and temperature, the enzyme activity reduced considerably to 0.0083, 0.0143 and 0.0037 U/mL at pH 9 and 0.0154, 0.0197 and 0.0121 U/mL at 50 °C by the tested bacterial strains. The same trend was also obtained in oth er substrate concentrations such as 1.0, 1.5 and 2.0 %. With in the four substrate concentrations, the endoglucanase enzyme activity was more in 1.5% concentration at the tested pH and temperatures. From the over all result, it was observed that, among the three bacterial strains, the enzyme activity was more in Cellulomonas sp, followed by Bacillus and Micrococcus spp. in varying pH and temperature.     

地下水中三氯乙烯-苯酚的好氧共代谢的实验研究

以苯酚作为三氯乙烯（TCE）降解的共代谢基质,用瓦勃氏微量呼吸测压仪（简称瓦呼仪）作为测试手段,分析了经苯酚驯化后的混合微生物对苯酚、TCE的降解特性;并讨论了以苯酚作为共代谢基质时TCE降解的可能性。实验结果表明：未驯化的活性污泥不能降解TCE; 经苯酚驯化后的活性污泥,当TCE的质量浓度为50 μg/L时其降解效果较好,TCE的氧化率达3369%;TCE的质量浓度为100 μg/L时其降解效果较差,其氧化率仅为3.2%;苯酚和TCE共代谢降解时,苯酚的存在促进了TCE的降解,当苯酚质量浓度为40 mg/L、TCE质量浓度为50 μg/L时共代谢降解效果最好,TCE的氧化率为79.11%。     

Effects of aeration rate and detention time on thermophilic aerobic digestion of mixed sludge and its dewaterability

Thermophilic aerobic digestion (TAD) is a possible alternative for rapid sludge degradation and producing Class A biosolids. Aeration rate and detention time are two of the important parameters in TAD processes due to rapidly growing thermophilic bacteria population, limited solubility of oxygen at high temperatures and the need to prevent cooling of TAD process. Also the current knowledge and understanding of dewatering TAD biosolids are limited and incompelet. The objectives of this study were to investigate the effects of various aeration rates and detention time on some characteristics of mixed sewage sludge. Four glass-cylindrical digesters with 7 liters of sludge in each, placed in a water bath and were operated at 55 °C with the aeration rates of 2.14, 3.00,3.86 and 4.71 volume of air per volume of sludge per hours. It was found the increase in aeration rate decreased the required detention time in order to meet the Class A sludge regulations to reduce vector attraction. The values of oxidation-reduction potential (ORP) were changed from negative values to positiv ones and the values of pH were increased from around neutral to slightly basic, but this increases were occured at different detention times. TAD affected dewaterability of mixed sewage sludge and produced biosolids with higher specific resistance to filtration (lower dewaterability) than undigested sludge. Thermophically digested sludge has a good setteling behavior and air drying on sand bed.     

Distribution and occurrence of aliphatic acid anions in deep subsurface waters

This study reports 144 new analyses of short-chain aliphatic acid anions (acetate, propionate, butyrate, and valerate) in formation waters from eight localities: Eastern Venezuelan Basin, Denver Basin, Eastern Green River Basin (Red Desert and Washakie Basins, Wamsutter Arch), San Juan Basin, Piceance Basin, Raton Basin, Gulf Coast Basin, and the Western Overthrust. Reservoir temperature does not predict total or relative abundance of aliphatic acid anions, but does predict maximum total concentrations of these species. Maximum concentrations increase to ~90°C. Above ~90°C, maximum concentrations decrease. Above ~250°C, maximum concentrations should not exceed ~1 mg/1. The general order of dominance is acetate > propionate > butyrate > valerate, but for coal-associated waters is propionate > acetate > butyrate > valerate. Lack of longer-chain aliphatic acid anion dominance over acetate at low reservoir temperatures may suggest hydrologie communication with deeper reservoirs.     

Fate and biodegradability potential of an emerging micropollutant diclofenac in subsurface environment

The present study deals with the assessment of the fate of an emerging contaminant (diclofenac), present in the subsurface environment at specific concentration levels, and its biodegradation potential under different environmental configurations. Diclofenac is a widely consumed nonsteroidal anti-inflammatory drug used as a painkiller prescribed as pills or ointments and among the most frequently detected pharmaceuticals in wastewater treatment plants’ effluents. Fermentative/methanogenic cultures were enriched from previously contaminated sediment samples. Although partial biodegradation was achieved in all enrichment cultures, complete degradation was not observed. The results reflected that less than 6% of the diclofenac sorbed on the solid phase and the rest was either biotransformed (25–40%) or remained in the liquid phase (55–70%) at the end of 45-day incubation period. Although partial cometabolic degradation of diclofenac occurred in the presence of glucose, biodegradation was not observed in the presence of readily biodegradable carbon source (i.e., acetate). Diclofenac concentration up to 1000 µg/L did not affect the methanogenic activity of the enriched culture. The results of this study will have a significant impact in the designation of the permitted concentration limits of diclofenac before the discharge through wastewater treatment plants.     

Analysis of the influence of free ammonia on the accumulation of nitrite in the nitrite nitrification system

Low- and high-concentration nitric wastewater can induce stable nitrite accumulation and realize the nitrification system, with the nitrite accumulation rate between 50% and 90%. In the low-concentration nitrite nitrification system, the average FA concentrations during the period of stable nitrite accumulation remained at 7 to 10 mg/L. In the high-concentration nitrite nitrification system, the nitrogen oxidation activity will not be affected by biomass, the nitrite oxidation activity is related to the system biomass, it is recommended to be measured by FA/MLSS. Keeping the FA sludge load below 0.1 is the precondition of making good nitrite accumulation and nitrogen degradation rate. Nitrite oxidation bacteria cannot be totally eliminated or washed out from the system.     

Ether lipids from the Lower and Middle Triassic at Qingyan,Guizhou Province,Southern China

Ether lipids such as non-isoprenoid mono and dialkyl glycerol ethers (MAGEs and DAGEs) and archaeol have been found in carbonate rocks and mud rocks deposited during the Early and Middle Triassic (250–240 Ma). The oldest previous discovery of ether lipids is from Cretaceous black shale deposited during the Albian (112 Ma). Paleoenvironmental reconstruction using ether lipids has therefore not been conducted on pre-Cretaceous sediments. Archaeol derives from archaea (e.g. methanogenic, methanotrophic, halophilic and thermophilic archaea). The non-isoprenoid DAGEs and MAGEs likely derive from sulfate-reducing bacteria (SRB), which are restricted to anaerobic environments. The presence of ether lipids thus indicates that anoxic conditions expanded in the depositional and/or water column environment in the Qingyan area during the Early–Middle Triassic. These ether lipids may be useful for the reconstruction of paleoredox conditions and paleoecosystems in the area. Furthermore, their occurrence suggests that paleoenvironmental reconstruction using ether lipids may be possible throughout the Mesozoic and Cenozoic.     

Bacterial and algal hydrocarbons in sediments from a saline Antarctic lake, Ace Lake

Capillary gas chromatography-mass spectrometry (C-GC-MS) and Iatroscan thin layer chromatography-flame ionisation detection (TLC-FID) were used to study hydrocarbon distributions in a sediment core from Ace Lake, a saline, meromictic lake in the Vestfold Hills of Antarctica. Hydrocarbons were abundant in most core sections (up to 125/μg/g dry wt), particularly in near-surface samples, and the distributions were very complex. Major constituents were identified as phytane, 2,6,10,15,19-pentamethyleicosane, tetrahydrosqualene, a mixture of phytenes, cholesta-3,5-diene and fern-7-ene. Smaller amounts of sterenes and hopenes were also present. The predominance of the first 3 acyclic isoprenoids in sediments buried less than 30 cm is consistent with high populations of methanogenic bacteria known to be present.Phytenes were abundant in all core sections, and there was no relationship between their abundance and that of phytane which suggests that they were not derived from methanogenic bacteria. Phytadienes were minor constituents at all depths studied. An unusual feature of some distributions was the high concentrations of fern-7-ene which was the major hydrocarbon in the 20–25 cm core section. This alkene was only abundant in sediments which contained high concentrations of methanogen markers suggesting that it may also be indicative of anoxic depositional environments. A possible source might be from purple non-sulphur bacteria. High concentrations of straight-chain C₂₉ and C₃₄ alkenes were also found in these sediments but their origin has not been determined. Major changes in the hydrocarbon distributions with depth indicate that the depositional environment in the lake has altered dramatically since the lake was formed less than 8000 years ago. The present condition of permanent anoxic bottom waters probably developed only in the last 1000 years.     

Plagioclase-aqueous solution equilibrium: Concentration dependence

The plagioclase-(NaCl + CaCl₂) exchange equilibrium was examined experimentally at 700°C, 0.5 GPa in aqueous solutions with salt concentrations from 1 to 64 m. The Ca/(Ca + Na) distribution between plagioclase and solution (salt melt) is illustrated in five diagrams constructed for concentrations of 1, 4, 8, 16, and 64 m. The elevated bulk salinity of the fluid at a constant Ca/(Ca + Na) ratio results in plagioclase albitization, with this effect reaching a maximum in relatively dilute solutions (1–4 m). In concentrated solutions (salt melts), the shift in the plagioclase composition with variations in the salinity is relatively insignificant. The simple hydration of basic rocks (purely metamorphic reaction) is associated with the albitization of plagioclase, and calculations suggest a possible shift from anorthite to oligoclase. This is also applicable to chemically more complex mineral associations: an increase in the overall salinity of the fluid should result in an increase in the activity of monovalent cations relative to that of bivalent ones and, correspondingly, stimulate reactions in which alkali earth cations (Ca + Mg + Fe) are substituted for alkalis (Na + K + Li). Although our experiments were carried out at temperatures 50°C lower than the melting point of albite under a pure water pressure (0.5 GPa), the addition of CaCl₂ solution to albite (i.e., plagioclase anorthitization and a decrease in the water activity in the salt solutions) induced the appearance of melt because of quartz formation by the reaction 2Ab + CaCl₂ → An + 2NaCl + 4Qtz and the eutectic phase proportions in the Ab + Qtz system.     

A geoenvironmental application of burned wastewater sludge ash in soil stabilization

This paper studied the use of burned sludge ash as a soil stabilizing agent. The sludge ash was obtained from a public wastewater treatment plant, and it was burned at 550 °C. Different percentages of burned sludge ash were mixed with three different types of clayey soil. A laboratory study consisting of Atterberg’s limits test, unconfined compressive strength test, standard proctor density test, and swelling pressure test were carried out on samples treated with burned sludge at different percentages by dry weight of the clayey soils. The results show that the addition of 7.5 % of the burned sludge ash by the dry weight of the soil will increase the unconfined compressive strength and maximum dry density and also decrease the swelling pressure and the swell potential of the soil. The addition of percentage higher than 7.5 % by dry weight of the soil decreases both the maximum dry density and the unconfined compressive strength; as a result it showed less effectiveness in stabilizing the soil. The conclusion of this research revealed that the burned sludge ash can be used as a promising material for soil stabilization.     

Impact of recharge water temperature on bioclogging during managed aquifer recharge: a laboratory study

To investigate the effect of recharge water temperature on bioclogging processes and mechanisms during seasonal managed aquifer recharge (MAR), two groups of laboratory percolation experiments were conducted: a winter test and a summer test. The temperatures were controlled at ~5±2 and ~15±3 °C, and the tests involved bacterial inoculums acquired from well water during March 2014 and August 2015, for the winter and summer tests, respectively. The results indicated that the sand columns clogged ~10 times faster in the summer test due to a 10-fold larger bacterial growth rate. The maximum concentrations of total extracellular polymeric substances (EPS) in the winter test were approximately twice those in the summer test, primarily caused by a ~200 μg/g sand increase of both loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS). In the first half of the experimental period, the accumulation of bacteria cells and EPS production induced rapid bioclogging in both the winter and summer tests. Afterward, increasing bacterial growth dominated the bioclogging in the summer test, while the accumulation of LB-EPS led to further bioclogging in the winter test. The biological analysis determined that the dominant bacteria in experiments for both seasons were different and the bacterial community diversity was ~50% higher in the winter test than that for summer. The seasonal inoculums could lead to differences in the bacterial community structure and diversity, while recharge water temperature was considered to be a major factor influencing the bacterial growth rate and metabolism behavior during the seasonal bioclogging process.     

The dominant acetate degradation pathway/methanogenic composition in full-scale anaerobic digesters operating under different ammonia levels

Ammonia is a major environmental factor influencing biomethanation in full-scale anaerobic digesters. In this study, the effect of different ammonia levels on methanogenic pathways and methanogenic community composition of full-scale biogas plants was investigated. Eight full-scale digesters operating under different ammonia levels were sampled, and the residual biogas production was followed in fed-batch reactors. Acetate, labelled in the methyl group, was used to determine the methanogenic pathway by following the ¹⁴CH₄ and ¹⁴CO₂ production. Fluorescence in situ hybridisation was used to determine the methanogenic communities’ composition. Results obtained clearly demonstrated that syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis was the dominant pathway in all digesters with high ammonia levels (2.8–4.57 g NH₄ ⁺-N L^?1), while acetoclastic methanogenic pathway dominated at low ammonia (<1.21 g NH₄ ⁺-N L^?1). Thermophilic Methanomicrobiales spp. and mesophilic Methanobacteriales spp. were the most abundant methanogens at free ammonia concentrations above 0.44 g NH₃-N L^?1 and total ammonia concentrations above 2.8 g NH₄ ⁺-N L^?1, respectively. Meanwhile, in anaerobic digesters with low ammonia (<1.21 g NH₄ ⁺-N L^?1) and free ammonia (<0.07 g NH₃-N L^?1) levels, mesophilic and thermophilic Methanosaetaceae spp. were the most abundant methanogens.     

微生物白云岩形成机理、识别标志及存在的问题

关于白云岩成因,前人提出了多种机理,但许多机理并没有过硬的证据,难以令人信服.近些年的研究表明,白云石借助微生物的作用在常温下可以沉淀出来,形成微生物白云岩.模拟实验证明了这种机理的正确性.微生物白云岩的发现表明,自然界中可以存在原生白云岩,因此具有重要科学意义.实验表明,多种微生物可形成白云岩.文章主要描述了其中4类...