Thermophilic aerobic wastewater treatment,process performance,biomass characteristics,and effluent quality

Thermophilic aerobic wastewater treatment is reviewed. Thermophilic processes have been studied in laboratory and pilot-scale while full-scale applications are rare. The paper focuses on the microbiology of aerobic thermophiles, performance of the aerobic wastewater treatments, sludge yield, and alternatives to enhance performance of thethermophilic process. Thermophilic processes have been shown to operate under markedly high loading rates (30–180 kg COD m⁻³d⁻¹).Reported sludge production values under thermophilic conditions vary between 0.05 and0.3 kg SS kg COD_removed, which are about the same or lower than generally obtained in mesophilic processes. Compared to analogous mesophilic treatment, thermophilic treatment commonly suffers from poorer effluent quality, measured by lower total COD and filtrated (GF-A) COD removals. However, in the removal of soluble (bacterial membrane filtered) COD both mesophilic and thermophilic treatments have produced similar results. Sludge settle ability in thermophilic processes have been reported to be better or poorer than in analogous mesophilic processes, although cases with better settling properties are rare. Combining thermophilic with mesophilic treatment or ultrafiltration may in some cases markedly improve effluent quality. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparison of static,in-vessel composting of MSW with thermophilic anaerobic digestion and combinations of the two processes

The biological stabilisation of the organic fraction of municipal solid waste (OFMSW) into a form stable enough for land application can be achieved via aerobic or anaerobic treatments. To investigate the rates of degradation (e.g. via electron equivalents removed, or via carbon emitted) of aerobic and anaerobic treatment, OFMSW samples were exposed to computer controlled laboratory-scale aerobic (static in-vessel composting), and anaerobic (thermophilic anaerobic digestion with liquor recycle) treatment individually and in combination. A comparison of the degradation rates, based on electron flow revealed that provided a suitable inoculum was used, anaerobic digestion was the faster of the two waste conversion process. In addition to faster maximum substrate oxidation rates, anaerobic digestion (followed by post-treatment aerobic maturation), when compared to static composting alone, converted a larger fraction of the organics to gaseous end-products (CO₂ and CH₄), leading to improved end-product stability and maturity, as measured by compost self-heating and root elongation tests, respectively. While not comparable to windrow and other mixed, highly aerated compost systems, our results show that in the thermophilic, in-vessel treatment investigated here, the inclusion of a anaerobic phase, rather than using composting alone, improved hydrolysis rates as well as oxidation rates and product stability. The combination of the two methods, as used in the DiCOM^® process, was also tested allowing heat generation to thermophilic operating temperature, biogas recovery and a low odour stable end-product within 19 days of operation.     

Bacterial community analysis of swine manure treated with autothermal thermophilic aerobic digestion

Due to the enviornmental problems associated with disposal of livestock sludge, many stabilization studies emphasizing on the sludge volume reduction were performed. However, little is known about the microbial risk present in sludge and its stabilized products. This study microbiologically explored the effects of anaerobic lagoon fermentation (ALF) and autothermal thermophilic aerobic digestion (ATAD) on pathogen-related risk of raw swine manure by using culture-independent 16S rDNA cloning and sequencing methods. In raw swine manure, clones closely related to pathogens such as Dialister pneumosintes, Erysipelothrix rhusiopathiae, Succinivibrioan dextrinosolvens, and Schineria sp. were detected. Meanwhile, in the mesophilic ALF-treated swine manure, bacterial community clones closely related to pathogens such as Schineria sp. and Succinivibrio dextrinosolvens were still detected. Interestingly, the ATAD treatment resulted in no detection of clones closely related to pathogens in the stabilized thermophilic bacterial community, with the predominance of novel Clostridia class populations. These findings support the superiority of ATAD in selectively reducing potential human and animal pathogens compared to ALF, which is a typical manure stabilization method used in livestock farms.     

Persistence of Mycobacterium avium subsp. paratuberculosis and other zoonotic pathogens during simulated composting, manure packing, and liquid storage of dairy manure

Livestock manures contain numerous microorganisms which can infect humans and/or animals, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis). The effects of commonly used manure treatments on the persistence of these pathogens have rarely been compared. The objective of this study was to compare the persistence of artificially inoculated M. paratuberculosis, as well as other naturally occurring pathogens, during the treatment of dairy manure under conditions that simulate three commonly used manure management methods: thermophilic composting at 55 degrees C, manure packing at 25 degrees C (or low-temperature composting), and liquid lagoon storage. Straw and sawdust amendments used for composting and packing were also compared. Manure was obtained from a large Ohio free-stall dairy herd and was inoculated with M. paratuberculosis at 10(6) CFU/g in the final mixes. For compost and pack treatments, this manure was amended with sawdust or straw to provide an optimal moisture content (60%) for composting for 56 days. To simulate liquid storage, water was added to the manure (to simulate liquid flushing and storage) and the slurry was placed in triplicate covered 4-liter Erlenmeyer flasks, incubated under ambient conditions for 175 days. The treatments were sampled on days 0, 3, 7, 14, 28, and 56 for the detection of pathogens. The persistence of M. paratuberculosis was also assessed by a PCR hybridization assay. After 56 days of composting, from 45 to 60% of the carbon in the compost treatments was converted to CO2, while no significant change in carbon content was observed in the liquid slurry. Escherichia coli, Salmonella, and Listeria were all detected in the manure and all of the treatments on day 0. After 3 days of composting at 55 degrees C, none of these organisms were detectable. In liquid manure and pack treatments, some of these microorganisms were detectable up to 28 days. M. paratuberculosis was detected by standard culture only on day 0 in all the treatments, but was undetectable in any treatment at 3 and 7 days. On days 14, 28, and 56, M. paratuberculosis was detected in the liquid storage treatment but remained undetectable in the compost and pack treatments. However, M. paratuberculosis DNA was detectable through day 56 in all treatments and up to day 175 in liquid storage treatments. Taken together, the results indicate that high-temperature composting is more effective than pack storage or liquid storage of manure in reducing these pathogens in dairy manure. Therefore, thermophilic composting is recommended for treatment of manures destined for pathogen-sensitive environments such as those for vegetable production, residential gardening, or application to rapidly draining fields.     

The Happy Factor treatment threshold,used to determine Targeted Selective Treatment decisions for lambs,is transferable between farms

Weight gain-based treatment decision-making has been shown to successfully reduce the number of anthelmintic treatments without compromising production as part of a Targeted Selective Treatment (TST)-based worm control strategy in sheep. The effects of using an efficiency threshold (Standard Threshold (ST)) developed on one farm were examined to establish whether there was a need to tailor the threshold for individual farm conditions. The Standard Threshold had been used on a number of farms, and data from these trials were used here. The ideal threshold (Estimated Treatment Threshold) for each farm was calculated using the same method as the original threshold, and the effect on the number of treatments given and subsequent productivity was estimated. Estimated treatment thresholds were calculated to be higher on all farms including the original, resulting in increased numbers of treatments due. The effect of the increased number of treatments was calculated to have no effect on productivity however, and it was concluded that the ST was sufficient, at least initially, for successful implementation of TST and that further refinement could be made using locally derived data if required.     

Discrimination of mesophilic and thermophilic proteins using machine learning algorithms

Discriminating thermophilic proteins from their mesophilic counterparts is a challenging task and it would help to design stable proteins. In this work, we have systematically analyzed the amino acid compositions of 3075 mesophilic and 1609 thermophilic proteins belonging to 9 and 15 families, respectively. We found that the charged residues Lys, Arg, and Glu as well as the hydrophobic residues, Val and Ile have higher occurrence in thermophiles than mesophiles. Further, we have analyzed the performance of different methods, based on Bayes rules, logistic functions, neural networks, support vector machines, decision trees and so forth for discriminating mesophilic and thermophilic proteins. We found that most of the machine learning techniques discriminate these classes of proteins with similar accuracy. The neural network-based method could discriminate the thermophiles from mesophiles at the five-fold cross-validation accuracy of 89% in a dataset of 4684 proteins. Moreover, this method is tested with 325 mesophiles in Xylella fastidosa and 382 thermophiles in Aquifex aeolicus and it could successfully discriminate them with the accuracy of 91%. These accuracy levels are better than other methods in the literature and we suggest that this method could be effectively used to discriminate mesophilic and thermophilic proteins.     

沼液的定价方法及其应用效果研究

为了探索沼液的商品化,采用养分含量定价和应用效果定价2种方法确定了沼液的价格,并分析了沼液施用后对土壤基本理化性状的影响作用。结果发现:采用养分含量定价法得到的沼液价格为78.12元/m³,采用沼液应用效果定价法得到的沼液的价格为111.4元/ m³;沼液的实际应用价值要明显高于其养分含量的市场价;沼液施用后可降低土壤容重,增加土壤孔隙度,增加土壤有机质、土壤全氮、土壤有效磷及有效钾含量,有利于土壤肥力的保持。     

Evaluation of methods for preparing hydrogen-producing seed inocula under thermophilic condition by process performance and microbial community analysis

Five methods for preparation of hydrogen-producing seeds (base, acid, 2-bromoethanesulfonic acid (BESA), load-shock and heat shock treatments) as well as an untreated anaerobic digested sludge were compared for their hydrogen production performance and responsible microbial community structures under thermophilic condition (60 degrees C). The results showed that the load-shock treatment method was the best for enriching thermophilic hydrogen-producing seeds from mixed anaerobic cultures as it completely repressed methanogenic activity and gave the a maximum hydrogen production yield of 1.96 mol H(2) mol(-1) hexose with an hydrogen production rate of 11.2 mmol H(2) l(-1)h(-1). Load-shock and heat-shock treatments resulted in a dominance of Thermoanaerobacterium thermosaccharolyticum with acetic acid and butyric acid type of fermentation while base- and acid-treated seeds were dominated by Clostridium sp. and BESA-treated seeds were dominated by Bacillus sp. The comparative experimental results from hydrogen production performance and microbial community analysis showed that the load-shock treatment method was better than the other four methods for enriching thermophilic hydrogen-producing seeds from anaerobic digested sludge. Load-shock treated sludge was implemented in palm oil mill effluent (POME) fermentation and was found to give maximum hydrogen production rates of 13.34 mmol H(2) l(-1)h(-1) and resulted in a dominance of Thermoanaerobacterium spp. Load-shock treatment is an easy and practical method for enriching thermophilic hydrogen-producing bacteria from anaerobic digested sludge.     

Persistence of Mycobacterium avium subsp. paratuberculosis and Other Zoonotic Pathogens during Simulated Composting, Manure Packing, and Liquid Storage of Dairy Manure

Livestock manures contain numerous microorganisms which can infect humans and/or animals, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis). The effects of commonly used manure treatments on the persistence of these pathogens have rarely been compared. The objective of this study was to compare the persistence of artificially inoculated M. paratuberculosis, as well as other naturally occurring pathogens, during the treatment of dairy manure under conditions that simulate three commonly used manure management methods: thermophilic composting at 55°C, manure packing at 25°C (or low-temperature composting), and liquid lagoon storage. Straw and sawdust amendments used for composting and packing were also compared. Manure was obtained from a large Ohio free-stall dairy herd and was inoculated with M. paratuberculosis at 10⁶ CFU/g in the final mixes. For compost and pack treatments, this manure was amended with sawdust or straw to provide an optimal moisture content (60%) for composting for 56 days. To simulate liquid storage, water was added to the manure (to simulate liquid flushing and storage) and the slurry was placed in triplicate covered 4-liter Erlenmeyer flasks, incubated under ambient conditions for 175 days. The treatments were sampled on days 0, 3, 7, 14, 28, and 56 for the detection of pathogens. The persistence of M. paratuberculosis was also assessed by a PCR hybridization assay. After 56 days of composting, from 45 to 60% of the carbon in the compost treatments was converted to CO₂, while no significant change in carbon content was observed in the liquid slurry. Escherichia coli, Salmonella, and Listeria were all detected in the manure and all of the treatments on day 0. After 3 days of composting at 55°C, none of these organisms were detectable. In liquid manure and pack treatments, some of these microorganisms were detectable up to 28 days. M. paratuberculosis was detected by standard culture only on day 0 in all the treatments, but was undetectable in any treatment at 3 and 7 days. On days 14, 28, and 56, M. paratuberculosis was detected in the liquid storage treatment but remained undetectable in the compost and pack treatments. However, M. paratuberculosis DNA was detectable through day 56 in all treatments and up to day 175 in liquid storage treatments. Taken together, the results indicate that high-temperature composting is more effective than pack storage or liquid storage of manure in reducing these pathogens in dairy manure. Therefore, thermophilic composting is recommended for treatment of manures destined for pathogen-sensitive environments such as those for vegetable production, residential gardening, or application to rapidly draining fields.     

Bioremediation of tetryl-contaminated soil using sequencing batch soil slurry reactor

A laboratory study was conducted to determine whether tetryl (2,4,6-trinitrophenlymethylnitramine) contaminated soil could be bioremediated using a sequencing batch soil slurry reactor (SBR) operated under anoxic–aerobic sequence. The results indicated that tetryl was co-metabolically converted to aniline under anoxic conditions with molasses as the growth substrate. The gas chromatographic/mass spectrometric analysis of the soil slurry showed various metabolites, identified as trinitrobenzeneamine, dintrobenzenediamine, nitroaniline and aniline. Aniline was not metabolized further under anoxic conditions. When the soil slurry reactor was operated under aerobic conditions, the aniline concentration was reduced to below the detection limit (0.05 ppm). This metabolic conversion of tetryl is probably of value in the treatment of tetryl-contaminated soil and ground water, such as those found at the Joliet army ammunition plant site in Illinois and the Iowa army ammunition plant site in Burlington, Iowa.     

The biosecurity of on-farm mortality composting

Composting is a natural biological decomposition process that takes place under aerobic and thermophilic conditions. It can be used for the day-to-day management of mortalities on farms and for carcass disposal in emergency animal disease (EAD) outbreaks. In mortality composting, carcasses are placed in piles or bins together with supplemental carbon sources such as sawdust, litter, straw or wood shavings. Composting is particularly suitable for broiler-farm mortalities and litter. In the case of emergency disease outbreaks, composting can be conducted either inside or outside the poultry house following killing. Composting has been successfully employed for emergency disposal of carcasses in a few cases in North America, but research is lacking on the biosecurity of the process. Composting is a well-established pathogen reduction technology, but process management and heterogenous pile conditions pose particular challenges for validating the microbiological safety of mortality composting. This paper reviews the available information on the biosecurity of mortality composting, identifies potential sources of risk, and highlights emerging research needs. Reports to date of the use of composting in EAD outbreaks are also discussed.     

Physiology and enzymology of thermophilic anaerobic bacteria degrading starch

Abstract The capability of secreting thermoactive enzymes exhibiting α-amylase and pullulanase with debraching activity, seems to be widely distributed amongst anaerobic thermophilic bacteria. Interestingly, pullulanase formed by these bacteria displays dual specificity by attacking α-1,6- as well as α-1,4-glycosidic linkages in branched glucose polymers. Unlike the enzyme system of aerobic microorganisms the majority of starch hydrolysing enzymes of anaerobic bacteria is metal indepedent and is extremely thermostable. This enzyme system is controlled by substrate induction and catabolite repression; enzyme expression is accomplished when maltose or maltose-containing carbohydrates are used as substrates. By developing a process in continuous culture we were able to greatly enhance enzyme synthesis and release by anaerobic thermophilic bacteria. An elevation in the specific activities of cell-free amylases and pullulanases could also be achieved by entrapping of bacteria in calcium alginate beads. The unique properties of extracellular enzymes of thermophilic anaerobic bacteria makes this group of organisms suitable candidates for inductrial application.     

Analysis of the bacterial community inhabiting an aerobic thermophilic sequencing batch reactor (AT-SBR) treating swine waste

The microflora of a self-heating aerobic thermophilic sequencing batch reactor (AT-SBR) treating swine waste was investigated by a combination of culture and culture-independent techniques. The temperature increased quickly in the first hours of the treatment cycles and values up to 72°C were reached. Denaturing gradient gel electrophoresis of the PCR-amplified V3 region of 16S rDNA (PCR-DGGE) revealed important changes in the bacterial community during 3-day cycles. A clone library was constructed with the near-full-length 16S rDNA amplified from a mixed-liquor sample taken at 60°C. Among the 78 non-chimeric clones analysed, 20 species (here defined as clones showing more than 97% sequence homology) were found. In contrast to other culture-independent bacterial analyses of aerobic thermophilic wastewater treatments, species belonging to the Bacilli class were dominant (64%) with Bacillus thermocloacae being the most abundant species (38%). The other Bacilli could not be assigned to a known species. Schineria larvae was the second most abundant species (14%) in the clone library. Four species were also found among the 19 strains isolated, cultivated and identified from samples taken at 40°C and 60°C. Ten isolates showed high 16S rDNA sequence homology with the dominant bacterium of a composting process that had not been previously isolated.An erratum to this article can be found at     

Thermophilic anaerobic digestion of Lurgi coal gasification wastewater in a UASB reactor

Lurgi coal gasification wastewater (LCGW) is a refractory wastewater, whose anaerobic treatment has been a severe problem due to its toxicity and poor biodegradability. Using a mesophilic (35 ± 2 °C) reactor as a control, thermophilic anaerobic digestion (55 ± 2 °C) of LCGW was investigated in a UASB reactor. After 120 days of operation, the removal of COD and total phenols by the thermophilic reactor could reach 50-55% and 50-60% respectively, at an organic loading rate of 2.5 kg COD/(m³ d) and HRT of 24 h; the corresponding efficiencies were both only 20-30% in the mesophilic reactor. After thermophilic digestion, the wastewater concentrations of the aerobic effluent COD could reach below 200 mg/L compared with around 294 mg/L if mesophilic digestion was done and around 375 mg/L if sole aerobic pretreatment was done. The results suggested that thermophilic anaerobic digestion improved significantly both anaerobic and aerobic biodegradation of LCGW.     

Temporal change of composition and potential activity of the thermophilic archaeal community during the composting of organic material

To date, composting has been regarded as an aerobic process but it has been shown that composting piles are often sources of atmospheric methane. In order to gain a more comprehensive view on the diversity of methanogenic Archaea in compost, gas chromatographical methods and molecular cloning were used to study relationships of thermophilic archaeal communities and changes in methane production potential during compost maturation. According to the thermophilic methane production potential, wide differences could be detected between differently aged compost materials. In material derived from 3- and 4-week-old piles, low and no thermophilic methane production potential, respectively, was observed at 50 degrees C. Material from a 6-week-old pile showed the maximum methane production. With compost maturation, the production slowly decreased again with 6 weeks, 8 weeks, and mature compost showing an optimum methane production potential at 60 degrees C. At 70 degrees C, only 6-week-old material showed a comparable high production of methane. The 16S rRNA-based phylogenetic surveys revealed an increase of archaeal diversity with compost maturation. In the 6-week-old material, 86% of the sequences in the archaeal 16S rRNA library had the highest sequence similarities to Methanothermobacter spp. and the remaining 14% of the clones were related to Methanosarcina thermophila. Quantification of methanogens in 6-week-old material, on the basis of the methane production rate, resulted in values of about 2x10(7) cells per gram fresh weight. In 8-week-old and mature compost material, the proportion of sequences similar to Methanothermobacter spp. decreased to 34% and 0%, respectively. The mature compost material showed the highest variation in identified sequences, although 33% could be assigned to as yet uncultured Archaea (e.g. Rice cluster I, III, and IV). Our results indicate that compost harbours a diverse community of thermophilic methanogens, with changing composition during the maturation process, presumably due to altered pile conditions. Likewise, compost may act as a potential carrier for thermophilic methanogens in temperate soils because it is widely used as a soil amendment.     

Process engineering in biological aerobic waste-water treatment

A non-comprehensive review of several technical developments in the field of aerobic biological waste-water treatment engineering is carried out, considering the active role the engineers have to play in this field. This paper brings together conventional and advanced problems in the field of aerobic biological waste-water treatment. Such an overview of biological waste-water treatment also precedes comments on some important aspects concerning the microorganisms responsible for waste-water treatment as well as considerations of the application of fundamentals and kinetics to the analysis of the biological processes used most commonly for aerobic biological waste-water treatment. A survey of the development of the biological activated-sludge process and some modifications are given. Some problems implied in the conventional activated-sludge waste-water treatment are analyzed, considering conventional processes and bioreactor models (the continuous stirred-tank reactor model and the plug-flow reactor models of the activated-sludge process) as well as aerated lagoons. Further, modifications of the activated-sludge process are presented. These include additional details on the bioreactor progress and applications, with emphasis on aspects concerning airlift bioreactors and their variants, deep-shaft bioreactors and reciprocating jet bioreactors which are considered as the third generation of bioreactors owing to their important advantages in design, operation and performance in waste-water treatment. Sequencing-batch reactors and aerobic digestion processes, including conventional aerobic digestion, high-purity oxygen digestion, thermophilic aerobic digestion and cryophylic aerobic digestion are also reviewed. Finally, some aspects regarding the operational factors that are involved in the selection of the reactor type are included.     

The occurrence of intestinal parasites in swine slurry and their removal in activated sludge plants

Thirteen intensive pig farms and two activated sludge treatment plants for pig slurry in north-western Spain were studied from April 2005 to June 2006 in order to evaluate the presence of enteric pathogens (Cryptosporidium, Giardia and helminths) and the efficiency with which they were removed. These parasites were present on 53%, 7% and 38% of the farms studied, respectively, with concentrations of 10(4)-10(5) oocysts per litre (/L) for Cryptosporidium, 10(3)cysts/L for Giardia and 10(2)-10(3) eggs/L for helminths. The overall removal of parasites in the pig slurry treatment plants ranged from 86.7% to over 99.99%. The results revealed a constant reduction at each stage of the treatment system, with activated sludge processes being the most effective treatment in reducing pathogens in pig slurry, 78-81% for Cryptosporidium oocysts and over 99.9% for helminth eggs. A heat drying procedure for sludge removed 4.3 log units of Cryptosporidium oocysts, demonstrating the excellent effectiveness of this treatment for reducing pathogens in sludge intended to be applied to land.     

Development of anaerobic digestion methods for palm oil mill effluent (POME) treatment

Palm oil mill effluent (POME) is a highly polluting wastewater that pollutes the environment if discharged directly due to its high chemical oxygen demand (COD) and biochemical oxygen demand (BOD) concentration. Anaerobic digestion has been widely used for POME treatment with large emphasis placed on capturing the methane gas released as a product of this biodegradation treatment method. The anaerobic digestion method is recognized as a clean development mechanism (CDM) under the Kyoto protocol. Certified emission reduction (CER) can be obtained by using methane gas as a renewable energy. This review aims to discuss the various anaerobic treatments of POME and factors that influence the operation of anaerobic treatment. The POME treatment at both mesophilic and thermophilic temperature ranges are also analyzed.     

Farm disinfectants select for cyclohexane resistance, a marker of multiple antibiotic resistance, in Escherichia coli

AIMS: The aim of this study was to determine if three classes of farm disinfectants were able to select for ciprofloxacin or cyclohexane tolerant [indicative of a multiple antibiotic resistance (MAR) phenotype] Escherichia coli and if cyclohexane-tolerant E. coli could be isolated from farms. METHODS AND RESULTS: Chicken slurry containing ca 1 : 99 ratio ciprofloxacin resistant : susceptible E. coli (10 different resistant strains examined) was treated for 24 h with each of the disinfectants and examined for survival of resistant : susceptible strains. Ciprofloxacin-sensitive (n=5) and -resistant (n=5) E. coli were grown with sublethal concentrations of the disinfectants and then plated to agar containing ciprofloxacin or overlaid with cyclohexane. Escherichia coli (n=389) isolated from farms were tested for cyclohexane tolerance. Minimum inhibitory concentrations (MIC) were determined against representative isolates and mutants. The disinfectants did not select for the ciprofloxacin-resistant E. coli in poultry slurry but following growth with each of the three disinfectants, higher numbers (P < or = 0.023) of cyclohexane-tolerant E. coli were isolated and these had a MAR phenotype. Of the 389 farm E. coli tested, only one was cyclohexane tolerant. CONCLUSIONS: It is possible that in a farm environment, E. coli could be exposed to similar concentrations of the disinfectants that are selected for MAR type organisms under these laboratory conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Data from this study suggest that cyclohexane-resistant E. coli are not common on farms, but in view of the ease of isolating them in the laboratory with farm disinfectants, further investigations on farms are warranted.