Synergy of two thermophiles enables decomposition of poly-epsilon-caprolactone under composting conditions

The ultimate degradation (i.e. complete mineralization) of biodegradable polymers proceeds through hydrolysis to the production of degradation intermediates (primary degradation) that are then taken into the microbial cell and further degraded to CO2 and water. We first isolated thermophilic actinomycete (Streptomyces thermonitrificans PDS-1), which has the activity of ultimate degradability, from compost in which poly-epsilon-caprolactone (PCL) degraded vigorously. We next tried to investigate the detailed mechanisms of degradation of the PCL in compost by developing a new experimental method in which isolated microorganisms are used to inoculate sterilized compost raw materials containing PCL. It was confirmed that the ultimate degradation of PCL could not be achieved by the action of the strain PDS-1 alone, and that a supplementary microorganism (Bacillus licheniformis HA1) isolated from compost utilizes the degradation intermediates and also increases the activity of the other primary microorganism (PDS-1) by adjusting the pH. We could thus show experimental proof of synergy between two thermophiles in the ultimate degradation of a biodegradable polymer in compost.     

Technological and economic potential of poly(lactic acid) and lactic acid derivatives

Abstract: Lactic acid has been an intermediate-volume specialty chemical (world production ∼ 40,000 tons/yr) used in a wide range of food processing and industrial applications. Lactic acid has the potential of becoming a very large volume, commodity-chemical intermediate produced from renewable carbohydrates for use as feedstocks for biodegradable polymers, oxygenated chemicals, plant growth regulators, environmentally friendly 'green' solvents, and specialty chemical intermediates. The recent announcements of new development-scale plants for producing lactic acid and polymer intermediates by major U.S. companies, such as Cargill, Ecochem (DuPont/ConAgra), and Archer Daniels Midland, attest to this potential.
In the past, efficient and economical technologies for the recovery and purification of lactic acid from crude fermentation broths and the conversion of lactic acid to the chemical or polymer intermediates had been the key technology impediments and main process cost centers. The development and deployment of novel separations technologies, such as electrodialysis (ED) with bipolar membranes, extractive distillations integrated with fermentation, and chemical conversion, can enable low-cost production with continuous processes in large-scale operations. The use of bipolar ED can virtually eliminate the salt or gypsum waste produced in the current lactic acid processes. Thus, the emerging technologies can use environmentally sound processes to produce environmentally useful products from lactic acid. The process economics of some of these processes and products can also be quite attractive. In this paper, the recent technical advances in lactic and polyactic acid processes are discussed. The economic potential and manufacturing cost estimates of several products and process options are presented. The technical accomplishments at Argonne National Laboratory (ANL) and the future directions of this program at ANL are discussed.     

THE EFFECTS OF MECHANICAL STIMULATION AND ETIOLATION ON THE COLLENCHYMA OF DATURA STRAMONIUM

Walker , Waldo S. (Grinnell College, Grinnell, Iowa.) The effects of mechanical stimulation and etiolation on the collenchyma of Datura stramonium. Amer. Jour. Bot. 47(9) : 717–724. Illus. 1960.–In an effort to determine the effect of mechanical stimulation on collenchyma tissue, plants of Datura stramonium L. were placed on a mechanical agitator and subjected to intensive shaking for 9 hr. per day for 40 days. Measurements indicated that such stimulation greatly increased the amount of wall thickening per cell, as observed in transverse section. Measurements also indicated that such stimulation may inhibit collenchyma cell elongation. A second group of Datura stramonium plants was placed in total darkness to determine the effect of such treatment on the quantity of wall thickening in the collenchyma tissue. Measurements indicated that when plants were placed in the dark for extended periods a great reduction of wall thickening resulted. It is suggested that reduction of wall material was due to its utilization as substrate for respiratory processes which occur in the plant under such extreme conditions. The composition and structure of the collenchyma cell walls are discussed.     

Assessing the effect of biodegradable and degradable plastics on the composting of green wastes and compost quality

An assessment of the effect of the composting potential of Mater-Bi biodegradable plastic with green wastes, noted by GBIO, and degradable plastic (PDQ-H additive) with green wastes, noted by GDEG, was carried out in a lagged two-compartment compost reactor. The composting time was determined until constant mass of the composting substrates was reached. The green wastes composting process was used as control (G). After one week of composting, the biodegradable plastics disappeared completely, while 2% of the original degradable plastic still remained after about 8 weeks of composting. A net reduction in volatile solids contents of 61.8%, 56.5% and 53.2% were obtained for G, GBIO and GDEG, respectively. Compost quality was assessed in terms of nitrogen, potassium and phosphorus contents, which were found to be highest for GBIO compost. From the phytotoxicity test, it has been observed that a diluted extract of GBIO compost has produced the longest length of radicle. From the respiration test, no significant difference in the amount of carbon dioxide released by the composting of GDEG and G was observed. This study showed that the quality of the compost is not affected by the presence of the biodegradable and degradable plastics in the raw materials.     

Bioremediation of 2,4,6-trinitrotoluene-contaminated soils by two different aerated compost systems

Two composting systems were compared on a laboratory scale as a bioredediation technology for degradation or immobilization of 2,4,6-trinitrotoluene (TNT) in contaminated soils. The first compost was aerated from the beginning whereas the second compost was only aerated after an anaerobic prephase of 65 days. In the first compost system the TNT concentration declined rapidly by 92% but, at the end, TNT could be partially recovered. During the anaerobic prephase of the second compost system, TNT was almost completely converted to aminodinitrotoluenes, which during the subsequent aeration almost entirely disappeared. In addition, the second compost generated less toxic material than the first one as confirmed by inhibition of bioluminescence ofVibrio fischeri. These data show that microbiological TNT-degradation systems can be successfully designed which are prerequisite for an efficient bioremediation of contaminated soils.     

Enhancement of the biological degradation of soils contaminated with oil by the addition of compost

The fundamentals of the biological treatment of contaminated soils were investigated in bioreactors with the aim to optimize the processes of biological soil treatment in order to achieve the highest possible degree of degradation within the shortest period of time. Preinvestigations using test systems at different scales have provided information about the possibilities of enhancing the decomposition processes which are dependent on various factors, such as milieu conditions, additives, etc., that must be known before remedial actions are taken. The investigations made so far have shown that compost is a favourable additive when oil-contaminated soils are biologically treated. The degradation of contaminants can be enhanced by the addition of compost. This positive effect is attributed to various mechanisms. In this paper, results of a variety of test systems at different scales are presented. In test series, different amounts of biocompost were added to investigate the influence on the degradation of diesel fuel. It was demonstrated that by increasing the compost content – the cumulative O₂ consumption caused by the degradation of the diesel fuel contaminants increased. It could be shown that the reduction of the diesel fuel contaminants in the soil was independent of the compost age and amounted to approximately 94% of the initial quantity. The addition of biocompost could also enhance the degradation of real contaminants. After a test period of 162 days in set-ups with compost addition, more than 75% of the lubricating oil contaminants disappeared, while less than 37% of the contaminants disappeared in set-ups without compost addition. Moreover, by the addition of compost, the formation of pellets during the dynamic treatment of soil materials could be reduced.     

Cutinolytic esterase activity of bacteria isolated from mixed-plant compost and characterization of a cutinase gene from Pseudomonas pseudoalcaligenes

The objective of the current study was to examine cutinolytic esterase (i.e., cutinase) activity by pseudomonads and bacteria isolated from mixed-plant compost. Approximately 400 isolates representing 52 taxa recovered from mixed-plant compost using cuticle baits, along with 117 pseudomonad isolates obtained from a culture collection (i.e., non-compost habitats), were evaluated. The ability of isolates to degrade the synthetic cutin polycaprolactone (PCL) was initially measured. Isolates from 23 taxa recovered from the compost degraded PCL. As well, isolates from 13 taxa of pseudomonads cleared PCL. Secondary screening measured esterase activity induced by the presence of apple cuticle using the chromogenic substrate p-nitrophenyl butyrate. Eighteen isolates representing four taxa (Alcaligenes faecalis , Bacillus licheniformis , Bacillus pumilus , and Pseudomonas pseudoalcaligenes) recovered from compost exhibited substantial esterase activity when grown with cuticle. In contrast, none of the pseudomonad isolates from the culture collection produced appreciable esterase activity. Although degradation of PCL was not correlated with esterase activity, isolates that were unable to degrade PCL failed to produce measureable esterase activities. Zymogram analysis indicated that the esterases produced by bacteria from compost ranged in size from 29 to 47 kDa. A gene from P. pseudoalcaligenes (cutA) was found to code for a cutin-induced esterase consisting of 302 amino acids and a theoretical protein size of 32 kDa. The enzyme was unique and was most closely related to other bacterial lipases (≤48% similarity).     

THE LEWIS CENTRAL SCHOOl SITE (13PW5):

Abstract

Due to strong Indian opposition and ambiguous state laws, Iowa archaeologists have found it impossible to investigate aboriginal cemetery areas in recent years. The accidental discovery of an Archaic ossuary at a construction site on the grounds of the Lewis Central School in southwestern Iowa provided a turning point in Indian-archaeologist relations which ultimately led to a resolution of the problem through the enactment of new sections in the Iowa legal code. This article traces the history of events leading up to the discovery of the Lewis Central School site (13PW5), and provides substantive data concerning materials recovered from that site. This is done to demonstrate that a considerable amount of information is obtainable through such studies and to provide archaeologists and physical anthropologists with a frame of reference to evaluate the results obtained. The paper concludes with a presentation of additions and changes in the Iowa legal code and a discussior, of the effects the Iowa precedent may have on anthropologists, Indians, and state officials elsewhere.     

Limitations of Thioglycolate Broth as a Sterility Test Medium for Materials Exposed to Gaseous Ethylene Oxide

Although ethylene oxide is a reliable sterilizer, the process may be limited by diffusion. Thus, situations may exist where microorganisms are protected from the sterilizing gas. It is possible that the exterior of a substance may be sterilized, whereas the interior is not. We investigated three general types of materials in which this limitation of diffusion could occur: the bore of glass and plastic tubing, the center of cotton balls, and plastic adhesive film/paper backing interface. These materials were contaminated as close to their geometric center as possible with Bacillus subtilis var. niger spores occluded in crystals of sodium chloride. After exposure of the contaminated materials (except aluminum foil) to ethylene oxide, thioglycolate broth (a standard sterility-test medium) indicated sterility, whereas Trypticase Soy Broth indicated nonsterility. It is likewise possible that aerobic microorganisms, surviving in or on material after exposure to dry heat or steam sterilization processes, would not be recovered by thioglycollate broth. Entrapped aerobic organisms will probably not grow out in the low oxygen tension zone of an anaerobic medium such as thioglycollate broth. It is recommended than an aerobic medium such as Trypticase Soy Broth be used concurrently with thioglycolate broth for sterility testing.     

Intrinsic Bacterial Contamination of a Commercial Iodophor Solution: Investigation of the Implicated Manufacturing Plant

After an outbreak of peritoneal infections attributed to intrinsic contamination of a poloxamer-iodine solution with Pseudomonas aeruginosa, the manufacturer of the contaminated solution permitted investigation and sampling of materials within the plant. Pseudomonas spp. were recovered from two different unopened lots of solution and from numerous water samples obtained at the plant. The isolates from water identical to those of an isolate recovered from Prepodyne solution (West-Agro Chemical Co., Inc., Westwood, Kans., manufactured for AMSCO Medical Products Div., Erie, Pa.) manufactured 1 month earlier at the same plant. P. aeruginosa was not recovered from incoming city water. P. aeruginosa was recovered from sterile water and poloxamer-iodine after 48 h of incubation in a plant polyvinyl chloride pipe. Scanning electron micrographs of polyvinyl chloride pipe used in the plant showed massive concentrations of rod-shaped and coccobacillary cells apparently embedded in interior deposits of the pipe. Manufacturers of iodophors should be aware that pipes or other surfaces colonized with bacteria may be a source of contamination of their products.     

Microbial degradation of polycyclic aromatic hydrocarbons in soils affected by the organic matrix of compost

This paper describes the degradation of naphthalene, phenanthrene, anthracene, fluoranthene, and pyrene in soil and soil/compost mixtures. Compost addition facilitated the degradation of 500 mg naphthalene/kg soil and 100 mg/kg each of other polycyclic aromatic hydrocarbons (PAH) within 25 days in soil systems with water contents below the water-holding capacity. By means of a humic acid extraction, it was demonstrated that the decrease of PAH concentrations after compost addition was not caused by a sorption to organic matter preventing PAH analysis. The enhanced PAH degradation was examined in a series of batch experiments with contaminated soil to evaluate whether the effect of compost addition is caused by the microorganisms of the compost itself, by the properties of the organic matrix of the compost material, or by water-soluble fertilising substances. The experiments revealed that the release of fertilising substances from the compost and the shift of soil pH brought about by the compost did not cause the stimulatory effect. The microorganisms inherent to the compost were also not necessary for the enhanced degradation. Sterilised compost was recolonised by soil microorganisms after a lagphase yielding a degradation activity similar to that of the non-sterilised compost. The presence of the solid organic matrix of the compost seemed to be essential for the enhanced degradation. The soil/compost microflora, which was separated from the organic matrix in liquid cultures, exhibited a much lower degrading activity than in the presence of the solid organic material.     

Variation in reproductive investment among and within populations of the scorpion Centruroides vittatus

Although information concerning variation among and within populations is essential to understanding an organism's life history, little is known of such variation in any species of scorpion. We show that reproductive investment by the scorpion Centruroides vittatus varied among three Texas populations during one reproductive season. Females from the Kickapoo population produced smaller offspring and larger litters than females from the Independence Creek or Decatur populations; this pattern remained when adjusting for among population variation in either female mass or total litter mass. Relative clutch mass (RCM) and within-litter variability in offspring mass (V*) did not differ among populations. Among-population variation may result from genetic differences or from phenotypically plastic responses to differing environments. Within populations, the interrelationships among reproductive variables were similar for Decatur and Independence Creek: females investing more in reproduction (measured by total litter mass, TLM) produced larger litters and larger offspring, and V* decreased with increased mean offspring mass (and with decreased litter size at Decatur). At Kickapoo, larger females produced larger litters and had larger TLM; females investing more in reproduction produced larger litters but not larger offspring. Within litter variability in offspring mass was not correlated with any reproductive variables in this latter population. These patterns may be explained by the fractional clutch hypothesis, the inability of females precisely to control investment among offspring or morphological constraints on reproduction.     

Effect of Medicago sativa L. and compost on organic and inorganic pollutant removal from a mixed contaminated soil and risk assessment using ecotoxicological tests

Several Gentle Remediation Options (GRO), e.g., plant-based options (phytoremediation), singly and combined with soil amendments, can be simultaneously efficient for degrading organic pollutants and either stabilizing or extracting trace elements (TEs). Here, a 5-month greenhouse trial was performed to test the efficiency of Medicago sativa L., singly and combined with a compost addition (30% w/w), to treat soils contaminated by petroleum hydrocarbons (PHC), Co and Pb collected at an auto scrap yard. After 5 months, total soil Pb significantly decreased in the compost-amended soil planted with M. sativa, but not total soil Co. Compost incorporation into the soil promoted PHC degradation, M. sativa growth and survival, and shoot Pb concentrations [3.8 mg kg^?1 dry weight (DW)]. Residual risk assessment after the phytoremediation trial showed a positive effect of compost amendment on plant growth and earthworm development. The O₂ uptake by soil microorganisms was lower in the compost-amended soil, suggesting a decrease in microbial activity. This study underlined the benefits of the phytoremediation option based on M. sativa cultivation and compost amendment for remediating PHC- and Pb-contaminated soils.     

Structural changes of plant residues during decomposition in a compost environment

The degradation of plant material during composting was investigated qualitatively by scanning electron microscopy (SEM) and quantitatively by chemical methods. Decomposition of Miscanthus (Miscanthus oogiformis L.), hemp (Cannabis sativa L.) and wheat (Triticum aestivum L.) straw was observed by placing litterbags containing these materials in compost piles. Hemp and Miscanthus straw were more stable than wheat straw, but the two materials differed in the way they were degraded despite similar chemical compositions. Hemp straw was broken down in more flexible structures compared to the rigid breakdown of Miscanthus straw. It was concluded that the anatomical arrangement of the tissue is just as important as the content of recalcitrant compounds in determining decomposition rate. Thus, when using composted plant materials as growing medium, the choice of material must depend not only on nutritional quality but also on structural quality. This study indicated that hemp material might be a good structural component in a compost to be used as a growing medium.     

以生活垃圾堆肥为基质的废弃物铺网草皮建植研究

以生活垃圾堆肥为基质,5种生活废弃物为铺网,对高羊茅、草地早熟禾及本特3种草坪植物的草皮培植与铺坪应用进行了研究.结果表明:(1)在草皮培植中,同种草坪植物在不同铺网草皮中的生长效应差异较大,其中早熟禾在草席与塑料纱网草皮中的综合性能表现较好;本特和高羊茅在竹席和草席中的表现较好.(2)草皮的刈割再生生长能力总体上与草皮培植时的生长状况密切相关,草种与基质、铺网间的协同性存在差异,生长协同性较好的草皮组合为早熟禾的草席草皮、本特的草席与塑料纱网草皮以及高羊茅的草席、纤维网袋与塑料纱网草皮.(3)5种生活废弃物铺网中,塑料薄膜在草皮培植中与各草种间的协同性均最差,其铺网草皮在铺坪应用中的再生生长能力也较差,可见塑料薄膜不适宜作为草皮铺网.     

Chemical Studies on Heated Starch/Glycine Model Systems

Sixty-three compounds were isolated and identified from heated starch and from heated starch with glycine. Starch or starch with glycine was heated in an Erlenmeyer flask at 290°C under a nitrogen stream. The volatiles formed during heating were entrained on a porous polymer (Porapack Q). Residual charred materials in the flasks were extracted with methylene chloride. The oily dark brown materials recovered from both the Porapack Q traps and the methylene chloride extracts were subjected to gas chromatography and gas chromatography/mass spectrometry. The compounds identified include 3 hydrocarbons, 11 ketones, 3 aldehydes, 24 aromatic compounds, 10 furans and 12 nitrogen-containing compounds. Formation mechanisms of some major products are proposed.     

Fate of 14C-labeled anthracene and hexadecane in compost-manured soil

Experiments were carried out to evaluate the impact of the addition of ripe compost on the degradation of two ¹⁴C-labeled hydrocarbon model compounds (anthracene and hexadecane) in soil. The addition of mature compost (20 % dry wt./dry wt.) stimulated significantly the disappearance of the extractable fraction of both compounds. With compost, 23 % of the labeled anthracene was transformed into ¹⁴CO₂ and 42 % was fixed to the soil matrix irreversibly. In the unsupplemented control reactor more than 88 % of the original anthracene could be recovered by either of two applied organic extraction procedures. The formation of non-extractable bound residues was less significant with [¹⁴C] hexadecane since only 21 % of the labeled carbon had become non-extractable after 103 days. The results presented show that compost could stimulate the depletion of hydrocarbons by either mineralization or the formation of unextractable bound residues (humification). The latter process might be a significant route of depletion in soil especially, for those hydrocarbons that are mineralized only slowly. The meaning of this finding for the assessment of soil bioremediation is discussed.These authors contributed equally to the presented work and should therefore both be considered as first authors     

Biocontained Carcass Composting for Control of Infectious Disease Outbreak in Livestock

Intensive livestock production systems are particularly vulnerable to natural or intentional (bioterrorist) infectious disease outbreaks. Large numbers of animals housed within a confined area enables rapid dissemination of most infectious agents throughout a herd. Rapid containment is key to controlling any infectious disease outbreak, thus depopulation is often undertaken to prevent spread of a pathogen to the larger livestock population. In that circumstance, a large number of livestock carcasses and contaminated manure are generated that require rapid disposal.Composting lends itself as a rapid-response disposal method for infected carcasses as well as manure and soil that may harbor infectious agents. We designed a bio-contained mortality composting procedure and tested its efficacy for bovine tissue degradation and microbial deactivation. We used materials available on-farm or purchasable from local farm supply stores in order that the system can be implemented at the site of a disease outbreak. In this study, temperatures exceeded 55°C for more than one month and infectious agents implanted in beef cattle carcasses and manure were inactivated within 14 days of composting. After 147 days, carcasses were almost completely degraded. The few long bones remaining were further degraded with an additional composting cycle in open windrows and the final mature compost was suitable for land application. Duplicate compost structures (final dimensions 25 m x 5 m x 2.4 m; L x W x H) were constructed using barley straw bales and lined with heavy black silage plastic sheeting. Each was loaded with loose straw, carcasses and manure totaling ~95,000 kg. A 40-cm base layer of loose barley straw was placed in each bunker, onto which were placed 16 feedlot cattle mortalities (average weight 343 kg) aligned transversely at a spacing of approximately 0.5 m. For passive aeration, lengths of flexible, perforated plastic drainage tubing (15 cm diameter) were placed between adjacent carcasses, extending vertically along both inside walls, and with the ends passed though the plastic to the exterior. The carcasses were overlaid with moist aerated feedlot manure (~1.6 m deep) to the top of the bunker. Plastic was folded over the top and sealed with tape to establish a containment barrier and eight aeration vents (50 x 50 x 15 cm) were placed on the top of each structure to promote passive aeration. After 147 days, losses of volume and mass of composted materials averaged 39.8% and 23.7%, respectively, in each structure.     

Characterization of different compost extracts using Fourier-transform infrared spectroscopy (FTIR) and thermal analysis

Compost extract or “compost tea” is a liquid extract of compost obtained by mixing compost and water for a defined period of time. Compost tea contains nutrients and a range of different organisms and is applied to the soil or directly to plants with the principal aim of suppressing certain plant diseases. In addition, the application of compost tea supplies nutrients and organic matter to the soil. Thermal analysis and Fourier transform infrared spectroscopy (FTIR) are two widely applied analytical techniques for establishing the stability of compost, and although numerous studies have evaluated the capacity of compost tea to suppress plant diseases, there are no studies employing these techniques to characterize compost-tea. For the present study, 12 compost extracts were produced under varying conditions in a purpose-built reactor. Two different composts, an stable compost produced from manure and an unstable compost produced from municipal solid waste, respectively, two aeration systems (aerated and non-aerated extracts) and three temperatures (10, 20 and 30°C) were used in these experiments. The extracts were freeze-dried and subsequently analysed, together with the two composts, by means of FTIR and thermal analysis. Extracts produced from high stability compost, independently of the conditions of aeration and temperature, showed very similar results. In contrast, differences among extracts produced from the unstable compost were more noticeable. However, the different conditions of aeration and temperature during the production of the extracts only explained partially these differences, since the transformations undergone by compost over the 3 months that the experiments lasted were also reflected in the composition of the extracts. In spite of everything, extraction process favoured the degradation of easily oxidizable organic matter, which was more abundant in unstable compost. This degradation was more intense for non-aerated processes, probably due to the longer duration of these (10 days) with respect to aerated extractions (2 days). The effect of temperature was not clear in these experiments, although high temperatures could increase micro organism activity and consequently favour the degradation of easily oxidizable organic matter.     

Microbes and microplastics: Community shifts along an urban coastal contaminant gradient

Plastic substrates introduced to the environment during the Anthropocene have introduced new pathways for microbial selection and dispersal. Some plastic-colonising microorganisms have adapted phenotypes for plastic degradation (selection), while the spatial transport (dispersal) potential of plastic colonisers remains controlled by polymer-specific density, hydrography and currents. Plastic-degrading enzyme abundances have recently been correlated with concentrations of plastic debris in open ocean environments, making it critical to better understand colonisation of hydrocarbon degraders with plastic degradation potential in urbanised watersheds where plastic pollution often originates. We found that microbial colonisation by reputed hydrocarbon degraders on microplastics (MPs) correlated with a spatial contaminant gradient (New York City/Long Island waterways), polymer types, temporal scales, microbial domains and putative cell activity (DNA vs. RNA). Hydrocarbon-degrading taxa enriched on polyethylene and polyvinyl chloride substrates relative to other polymers and were more commonly recovered in samples proximal to New York City. These differences in MP colonisation could indicate phenotypic adaptation processes resulting from increased exposure to urban plastic runoff as well as differences in carbon bioavailability across polymer types. Shifts in MP community potential across urban coastal contaminant gradients and polymer types improve our understanding of environmental plastic discharge impacts toward biogeochemical cycling across the global ocean.