Dynamic of functional microbial groups during mesophilic composting of agro-industrial wastes and free-living (N2)-fixing bacteria application

Although several reports are available concerning the composition and dynamics of the microflora during the composting of municipal solid wastes, little is known about the microbial diversity during the composting of agro-industrial refuse. For this reason, the first parts of this study included the quantification of microbial generic groups and of the main functional groups of C and N cycle during composting of agro-industrial refuse. After a generalized decrease observed during the initial phases, a new bacterial growth was observed in the final phase of the process. Ammonifiers and (N₂)-fixing aerobic groups predominated outside of the piles whereas, nitrate-reducing group increased inside the piles during the first 23 days of composting. Ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), showed an opposite trend of growth since ammonia oxidation decreased with the increase of the nitrite oxidation activity. Pectinolytics, amylolytics and aerobic cellulolytic were present in greater quantities and showed an upward trend in both the internal and external part of the heaps.Several free-living (N₂)-fixing bacteria were molecularly identify as belonging especially to uncommon genera of nitrogen-fixing bacteria as Stenotrophomonas, Xanthomonas, Pseudomonas, Klebsiella, Alcaligenes, Achromobacter and Caulobacter. They were investigated for their ability to fix atmospheric nitrogen to employ as improvers of quality of compost. Some strains of Azotobacter chrococcum and Azotobacter salinestris were also tested. When different diazotrophic bacterial species were added in compost, the increase of total N ranged from 16% to 27% depending on the selected microbial strain being used. Such microorganisms may be used alone or in mixtures to provide an allocation of plant growth promoting rhizobacteria in soil.     

Microbial communities and greenhouse gas emissions associated with the biodegradation of specified risk material in compost

Provided that infectious prions (PrP^Sc) are inactivated, composting of specified risk material (SRM) may be a viable alternative to rendering and landfilling. In this study, bacterial and fungal communities as well as greenhouse gas emissions associated with the degradation of SRM were examined in laboratory composters over two 14 day composting cycles. Chicken feathers were mixed into compost to enrich for microbial communities involved in the degradation of keratin and other recalcitrant proteins such as prions. Feathers altered the composition of bacterial and fungal communities primarily during the first cycle. The bacterial genera Saccharomonospora, Thermobifida, Thermoactinomycetaceae, Thiohalospira, Pseudomonas, Actinomadura, and Enterobacter, and the fungal genera Dothideomycetes, Cladosporium, Chaetomium, and Trichaptum were identified as candidates involved in SRM degradation. Feathers increased (P < 0.05) headspace concentrations of CH₄ primarily during the early stages of the first cycle and N₂O during the second. Although inclusion of feathers in compost increases greenhouse gas emissions, it may promote the establishment of microbial communities that are more adept at degrading SRM and recalcitrant proteins such as keratin and PrP^Sc.     

Analysis of bacterial activity, biomass and diversity during windrow composting

Two contrasting compost windrows were monitored for various physical, chemical and microbiological parameters for a period of 106 days. The different input materials and management practises gave rise to different temperature, moisture, and oxygen consumption profiles as composting proceeded. However, despite the different composting conditions, the specific respiratory activity, as determined by oxygen consumption per bacterial cell, was remarkably similar for both windrows. Further investigations into diversity dynamics were done through DGGE and cloning and sequencing of bacterial 16S rDNA PCR products. Although sequence analysis showed differing bacterial communities across time and between the different windrows, similarities in the progression were noted. The majority of sequences recovered from the first sampling period (day 1) were highly similar to previously isolated organisms. The clone libraries from the last sampling period (day 106) contained organisms that showed lower homology to their closest relatives, often with other uncultured organisms, and in phyla that contain few cultured representatives. These data suggest that specific respiratory activity may be an important driver of bacterial diversity in composting environments.     

Coffee husk composting: An investigation of the process using molecular and non-molecular tools

Various parameters were measured during a 90-day composting process of coffee husk with cow dung (Pile 1), with fruit/vegetable wastes (Pile 2) and coffee husk alone (Pile 3). Samples were collected on days 0, 32 and 90 for chemical and microbiological analyses. C/N ratios of Piles 1 and 2 decreased significantly over the 90 days. The highest bacterial counts at the start of the process and highest actinobacterial counts at the end of the process (Piles 1 and 2) indicated microbial succession with concomitant production of compost relevant enzymes. Denaturing gradient gel electrophoresis of rDNA and COMPOCHIP microarray analysis indicated distinctive community shifts during the composting process, with day 0 samples clustering separately from the 32 and 90-day samples. This study, using a multi-parameter approach, has revealed differences in quality and species diversity of the three composts.     

Effect of temperature on bacterial emissions in composting of swine manure

Swine manure was subjected to laboratory scale composting in order to quantify bioaerosols, i.e., airborne culturable bacteria and endotoxin, in the exhaust gas, which provided details on the effect of temperature on bacterial emissions. The concentration of airborne bacteria reached 31,250 colony-forming units (CFU)/m³ during the thermophilic stage of composting, and positively correlated with the temperature profile of the compost pile. Initially, the endotoxin concentration was 1820 endotoxin units (EU)/m³, but it decreased exponentially as the composting process proceeded. The temperature can be an excellent indicator of bacterial emissions during the composting process, indicating that the composting process requires a consistently high temperature to ensure sanitization of both compost and bacterial emissions. The cumulative emission data showed that emission factors was 11.2?13.5 CFU/g dry swine manure and that of endotoxin was 0.5?0.9 EU/g dry swine manure. The bacterial diversity in the bioaerosol was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis, revealing the presence of various gram-negative bacterial consortia.     

Humification index of composts originating from three types of woody biomass

Composting is a good method for recycling surplus manure and stabilizing organic matter from biowastes. Compost is used as a soil amendment and recently, for restoration of vegetation in barren areas. We investigated the relationship between the type of woody biomass (using Robinia pseudoacacia, Japanese larch and apple) and the humification index (HI) of the resulting compost. This study evaluated the difference in HI between the three compost types, and the structural features of composts and extracted humic acids (HAs). The HIs for R. pseudoacacia and apple were larger than that for Japanese larch after composting for 11 months. The structural features of the Japanese larch compost were also different from the apple and R. pseudoacacia, with a very high carbon/nitrogen ratio. The average molecular weights and ultraviolet–visible spectra (A600/C) of HAs extracted from composting samples at 0 and 11 months indicate that the humification rate of Japanese larch was slower than that of R. pseudoacacia and apple. During composting, the average molecular weights of apple and R. pseudoacacia decreased, while their A600/C values increased, but the reverse was observed for Japanese larch. The humification rate was found to depend on the type of woody biomass being composted.     

Bacterial composting of animal fleshing generated from tannery industries

Animal fleshing (ANFL) is the major proteinaceous solid waste generated during the manufacture of leather, which requires to be disposed of by environmentally sound manner. This study reports about the treatment of ANFL into an organic compost and its effects on physiological parameters of different crops in a laboratory study. The ANFL was hydrolysed using Selenomonas ruminantium HM000123 and then the hydrolysed ANFL was mixed with cow dung and leaf litter for producing composted organic fertilizer (COF). The COF was characterized for pH, electrical conductivity (EC), total Kjeldhal nitrogen (TKN), and total organic carbon (TOC). The composting resulted in a significant reduction in pH, TOC and C:N ratio and an increase in TKN after 49 days in a compost reactor. Scanning electron microscope and FT-IR were used to analyse the hydrolysis of intra structural ANFL matrix and changes in the functional groups, respectively, in initial and final day COF. Thermogravimetry (TG) analysis was carried out for the raw mixture and COF samples to identify the weight loss under the nitrogen environment. The relative seed germination was found to be 94% in tomato (Lycopersicon esculentum), 92% in green gram (Vigna radiata), 86% in bottle gourd (Lagenaria siceraria (Mol.) Standl.) and 84% in cucumber (Cucumis sativus L.) using the extracts of COF. The results indicate that the combination of both hydrolysis and bacterial composting reduced the overall time required for composting and producing a nutrient-enriched compost product.     

Workers' exposure to dust, endotoxin and β-(1-3) glucan at four large-scale composting facilities

Objectives

To characterise compost workers’ exposure to dust, endotoxin and β-(1-3) glucan during various operational practices and investigate whether dust concentrations are a useful indicator of endotoxin exposure in compost workers.

Methods

This study assessed inhalable dust fractions, bacterial endotoxin and β-(1-3) glucan in 117 personal samples and 88 ambient samples from four large-scale composting facilities.

Results

Employees’ exposures to inhalable dust, endotoxin and β-(1-3) glucan exhibited a large range. Inhalable dust was found to be generally low (GM 0.99 mg/m³, GSD 2.99 mg/m³). Analysis of the biological component of the dust showed that employees’ exposures to endotoxin were elevated (GM 35.10 EU/m³, GSD 9.97 EU/m³). Employees’ exposure to β-(1-3) glucan was low (GM 0.98 ng/m³, GSD 13.39 ng/m³). Dust levels were elevated during manual sorting and screening of waste and high levels of endotoxin and β-(1-3) glucan were observed during all practices involving the movement of waste. A significant correlation was observed between the personal dust levels and personal endotoxin concentrations (r = 0.783, p < 0.05) and that personal inhalable dust concentration may be a valuable predictor for personal endotoxin concentration in the sites studied.

Conclusions

Workers at composting sites are exposed to high levels of bacterial endotoxin consistent with adverse respiratory outcomes even though in most cases, their personal dust exposure is below the suggested regulatory levels. Dose-response data for the biological components present in the dust encountered at composting sites are not well established at this time and site operators should adopt precautionary measures when assessing and managing these potential risks.     

Chemical,physical and microbial properties and microbial diversity in manufactured soils produced from co-composting green waste and biosolids

The effects of adding biosolids to a green waste feedstock (100% green waste, 25% v/v biosolids or 50% biosolids) on the properties of composted products were investigated. Following initial composting, 20% soil or 20% fly ash/river sand mix was added to the composts as would be carried out commercially to produce manufactured soil. Temperatures during composting reached 50 °C, or above, for 23 days when biosolids were included as a composting feedstock but temperatures barely reached 40 °C when green waste alone was composted. Addition of biosolids to the feedstock increased total N, EC, extractable NH₄, NO₃ and P but lowered pH, macroporosity, water holding capacity, microbial biomass C and basal respiration in composts. Additions of soil or ash/sand to the composts greatly increased the available water holding capacity of the materials. Principal component analysis (PCA) of PCR-DGGE 16S rDNA amplicons separated bacterial communities according to addition of soil to the compost. For fungal ITS-RNA amplicons, PCA separated communities based on the addition of biosolids. Bacterial species richness and Shannon’s diversity index were greatest for composts where soil had been added but for fungal communities these parameters were greatest in the treatments where 50% biosolids had been included. These results were interpreted in relation to soil having an inoculation effect and biosolids having an acidifying effect thereby favouring a fungal community.     

CIELAB color variables as indicators of compost stability

The composting process of different organic wastes both in laboratory and on a large-scale was characterized using CIELAB color variables to evaluate compost stability for the better application in agriculture. The time courses of the CIELAB variables of composting materials were determined directly from the bottom of a glass petri dish filled with dried and ground samples using a Minolta Color Reader (CR-13) calibrated with clean empty petri dishes placed on a white tile. To compare the proposed method with conventional methods, the same materials were also evaluated using commonly used compost stability evaluation indices. Most of the CIELAB variables of a compost made from a mixture of green tea waste and rice bran reached a plateau after 84 days of composting and showed strong relationships with the commonly used compost stability evaluation indices. The time needed for CIELAB variables, especially the L*and b* values, to stabilize at large-scale composting plants of cattle litter, farmyard manure, kitchen garbage and bark compost, were more or less similar to the times of maturation evaluated by the respective compost producers. The CIELAB color variable offers a new, simple, rapid and inexpensive means of evaluating compost stability and its quality prior to agricultural use.     

Production of well-matured compost from night-soil sludge by an extremely short period of thermophilic composting

The effect of various operational conditions on the decomposition of organic material during the composting of night-soil treatment sludge was quantitatively examined. The optimum composting conditions were found to be a temperature of ca. 60 °C and an initial pH value of 8. Rapid decomposition of organic matter ceased by the sixth day of composting under these optimum conditions, and the final value of the cumulative emission of carbon (E_C), which represents the degree of organic matter decomposition, was less than 40%, indicating that the sludge contained only a small amount of easily degradable organic material. A plant growth assay using Komatsuna (Brassica campestris L. var. rapiferafroug) in a 1/5000a standard cultivation pot was then conducted for the compost at various degrees of organic matter decomposition: the raw composting material, the final compost obtained on day 6, and the 2 intermediate compost products (i.e., E_C = 10% and 20%). It was found that the larger the E_C, the greater the yield of Komatsuna growth. It was also found that 6 days of composting is sufficient to promote Komatsuna growth at the standard loading level, which is equivalent to a 1.5 g N/pot, since the promotion effect was as high as that obtained using chemical fertilizer. It can therefore be concluded that well-matured compost could be obtained in a short period of time (i.e., as early as 6 days), when night-soil sludge is composted under optimum conditions.     

Vermistabilization and nutrient enhancement of anaerobic digestate through earthworm species Perionyx excavatus and Perionyx sansibaricus

The aim of the study was to examine safe reuse and recycling of organic waste digestate obtained from a biogas plant (5 % total solid) with enhanced nutrient value through vermitechnology. Two indigenous epigeic earthworm species Perionyx excavates and Perionyx sansibaricus were tried individually for this purpose. The results clearly show a significant increase in pH values from 6.5–7.4, electrical conductivity (21.3–21.7 %), total N (84.5–94.6 %), total P (35.9–47.1 %), total K (49.8–52.6 %), Ca (41.9–41.9 %) and a significant decrease in total organic C (17.1–22.4 %), C/N ratio (7.2–6.9), C/P ratio (20.3–20.6), COD (51.9–55.7 %), BOD (85.5–91.2 %). Similarly, indicator organisms (fecal coliforms and fecal streptococci) showed decrement at the end of the composting period (60 days). Fecal Coliforms reduce to nil, while in fecal streptococci a 6 log reduction was observed. Oxygen uptake rate dropped to 67.4–70 % for vermireactors. Overall, the aforementioned findings highlighted that the indigenous earthworm species could enhance the nutrient value of the anaerobic digestate, which could be utilized as an efficient soil conditioner.     

Effects of pH and microbial composition on odour in food waste composting

A major problem for composting plants is odour emission. Slow decomposition during prolonged low-pH conditions is a frequent process problem in food waste composting. The aim was to investigate correlations between low pH, odour and microbial composition during food waste composting. Samples from laboratory composting experiments and two large scale composting plants were analysed for odour by olfactometry, as well as physico-chemical and microbial composition. There was large variation in odour, and samples clustered in two groups, one with low odour and high pH (above 6.5), the other with high odour and low pH (below 6.0). The low-odour samples were significantly drier, had lower nitrate and TVOC concentrations and no detectable organic acids. Samples of both groups were dominated by Bacillales or Actinobacteria, organisms which are often indicative of well-functioning composting processes, but the high-odour group DNA sequences were similar to those of anaerobic or facultatively anaerobic species, not to typical thermophilic composting species. High-odour samples also contained Lactobacteria and Clostridia, known to produce odorous substances. A proposed odour reduction strategy is to rapidly overcome the low pH phase, through high initial aeration rates and the use of additives such as recycled compost.     

Indicator organisms for assessing sanitization during composting of plant wastes

The potential for using plant pathogens and seeds as indicator organisms for assessing sanitization of plant wastes during composting was tested in bench-scale flask and large-scale systems. Plasmodiophora brassicae was unsuitable due to high temperature tolerance in dry to moist composts, and detection of viable inoculum post-composting using bioassay plants not corresponding with that using TaqMan® PCR, possibly due to preservation of nucleic acids at elevated temperatures. Several other plant pathogens (Sclerotinia sclerotiorum, Microdochium nivale, Phytophthora cinnamomi and Phytophthora nicotianae) were unsuitable due their low temperature tolerance. Fusarium oxysporum f.sp. cepae and f.sp. radicis-lycopersici chlamydospores and tomato seeds were suitable indicators due to their moderate temperature tolerance and ease of viability testing post-composting. Abutilon seeds were more tolerant than tomato seeds of compost temperatures ?52 °C but more prone to degradation at lower temperatures and therefore less suitable as indicators. Relationships between compost temperature during exposures of 2-10 days and subsequent viability of the above chlamydospores or seeds enabled the sanitizing effect of composting processes to be predicted within 2-6 days. Plant waste type (woody or vegetable) had a small but significant effect on the relationship for tomato seeds but not for F. oxysporum chlamydospores.     

PLA and Organoclays Nanocomposites: Degradation Process and Evaluation of ecotoxicity Using Allium cepa as Test Organism

In this study, nanocomposites of PLA and organoclays Cloisite 20A and Cloisite 30B were prepared by the melt intercalation method and the obtained samples were characterized by transmission electron microscopy (TEM). Since composting is an important proposal to the final disposal of biopolymers, the influence of clays on the hydrolytic degradation process of PLA was evaluated by visual analysis and monitoring of molecular weight after periods of 15 and 30 days of degradation in compost. After degradation of the materials in composting environment, the evaluation of cytotoxic, genotoxic and mutagenic effects of compost aqueous extract was carried out using a bioassay with Allium cepa as test organism. The TEM micrographs permitted the observation of different levels of dispersion, including exfoliated regions. In the evaluation of hydrolytic degradation it was noted that the presence of organoclays can decrease the rate of degradation possibly due to the barrier effect of clay layers and/or the higher degree of crystallinity in the nanocomposite samples. Nevertheless, even in the case of nanocomposites, the molecular weight reduction was significant, indicating that the composting process is favorable to the chain scission of PLA in studied materials. In the analysis performed by the bioassay using A. cepa as test organism, it was found that after degradation of the PLA and its nanocomposites the aqueous extract of compost samples induced a decreasing in the mitotic index and an increasing in the induction of chromosomal abnormalities. These results were statistically significant in relation to the negative control (distilled water). By comparing the results obtained for the nanocomposites in relative to pure polymer, there were no statistically significant differences. The types of the observed chromosomal aberrations indicated a possible genotoxic effect of the materials, which may be related to an aneugenic action of PLA degradation products.     

Chemical characterization of emissions from a municipal solid waste treatment plant

Gaseous emissions are an important problem in municipal solid waste (MSW) treatment plants. The sources points of emissions considered in the present work are: fresh compost, mature compost, landfill leaks and leachate ponds. Hydrogen sulphide, ammonia and volatile organic compounds (VOCs) were analysed in the emissions from these sources. Hydrogen sulphide and ammonia were important contributors to the total emission volume. Landfill leaks are significant source points of emissions of H₂S; the average concentration of H₂S in biogas from the landfill leaks is around 1700 ppmv. The fresh composting site was also an important contributor of H₂S to the total emission volume; its concentration varied between 3.2 and 1.7 ppmv and a decrease with time was observed. The mature composting site showed a reduction of H₂S concentration (<0.1 ppmv). Leachate pond showed a low concentration of H₂S (in order of ppbv). Regarding NH₃, composting sites and landfill leaks are notable source points of emissions (composting sites varied around 30–600 ppmv; biogas from landfill leaks varied from 160 to 640 ppmv).Regarding VOCs, the main compounds were: limonene, p-cymene, pinene, cyclohexane, reaching concentrations around 0.2–4.3 ppmv.H₂S/NH₃, limonene/p-cymene, limonene/cyclohexane ratios can be useful for analysing and identifying the emission sources.     

Temperature and deactivation of microbial faecal indicators during small scale co-composting of faecal matter

Small scale co-composting of faecal matter from dry toilet systems with shredded plant material and food waste was investigated in respect to heat development and deactivation of faecal indicators under tropical semiarid conditions. Open (uncovered) co-composting of faecal matter with shredded plant material alone did not generate temperatures high enough (<55 °C) to reduce the indicators sufficiently. The addition of food waste and confinement in chambers, built of concrete bricks and wooden boards, improved the composting process significantly. Under these conditions peak temperatures of up to 70 °C were achieved and temperatures above 55 °C were maintained over 2 weeks. This temperature and time is sufficient to comply with international composting regulations. The reduction of Escherichia coli, Enterococcus faecalis and Salmonella senftenberg in test containment systems placed in the core of the compost piles was very efficient, exceeding 5 log₁₀-units in all cases, but recolonisation from the cooler outer layers appeared to interfere with the sanitisation efficiency of the substrate itself. The addition of a stabilisation period by extending the composting process to over 4 months ensured that the load of E. coli was reduced to less than 10³ cfu^?g and salmonella were undetectable.     

Fish Stomachs as a Biomonitoring Tool in Studies of Invertebrate Recovery

High mortality rates due to predation from fish may reduce densities of preferred prey animals. Predation may also depress the rate of recovery from environmental stress. In an alpine ecosystem damaged by acidification, we compared three different techniques of monitoring the recovery of two large species of crustaceans, the amphipod Gammarus lacustris and the notostrachan Lepidurus arcticus. The methods used were: (1): benthic littoral kick samples, (2): artificial substrate in the form of jute bags, (3): examination of brown trout stomachs. The monitoring took place in two limed lakes at the Hardangervidda mountain plateau in Central Norway, L. Svartavatn and L. Svartavasstjørni. Brown trout, Salmo trutta, is the only fish species in the lakes. Liming as a water quality improvement measure was started in 1994. All stomach samples were negative with respect to Gammarus and Lepidurus during the period 1987–1998. In 1999, the first records of both species were done in trout stomachs collected from Lake Svartavatnet. In Lake Svartavasstjørni, Lepidurus and Gammarus reappeared in fish stomachs in 2001 and 2002, respectively. During the period of monitoring, no records of these crustaceans were done in benthic samples and on artificial substrate in any of the two lakes. In an unacidified reference site, Lake Skiftesjøen, both benthic samples and the jute bags indicated a dense population of Gammarus. Our results strongly indicate that studies of fish stomachs are the best method for monitoring low-density populations of attractive fish food animals.     

Microbial diversity and dynamics during methane production from municipal solid waste

The objectives of this study were to characterize development of bacterial and archaeal populations during biodegradation of municipal solid waste (MSW) and to link specific methanogens to methane generation. Experiments were conducted in three 0.61-m-diameter by 0.90-m-tall laboratory reactors to simulate MSW bioreactor landfills. Pyrosequencing of 16S rRNA genes was used to characterize microbial communities in both leachate and solid waste. Microbial assemblages in effluent leachate were similar between reactors during peak methane generation. Specific groups within the Bacteroidetes and Thermatogae phyla were present in all samples and were particularly abundant during peak methane generation. Microbial communities were not similar in leachate and solid fractions assayed at the end of reactor operation; solid waste contained a more abundant bacterial community of cellulose-degrading organisms (e.g., Firmicutes). Specific methanogen populations were assessed using quantitative polymerase chain reaction. Methanomicrobiales, Methanosarcinaceae, and Methanobacteriales were the predominant methanogens in all reactors, with Methanomicrobiales consistently the most abundant. Methanogen growth phases coincided with accelerated methane production, and cumulative methane yield increased with increasing total methanogen abundance. The difference in methanogen populations and corresponding methane yield is attributed to different initial cellulose and hemicellulose contents of the MSW. Higher initial cellulose and hemicellulose contents supported growth of larger methanogen populations that resulted in higher methane yield.