Effects of the humic substances of de-inking paper sludge on the antagonism between two compost bacteria and Pythium ultimum

We investigated the in vitro influence of humic substances (HS) extracted from de-inking paper sludge compost on the inhibition of Pythium ultimum by two compost bacteria, Rhizobium radiobacter (Agrobacterium radiobacter) and Pseudomonas aeruginosa. When low concentrations (5 or 50 mg l(-1)) of HS were added to the culture medium, fungal inhibition by R. radiobacter significantly increased (P<0.01) by 2-3%. In contrast, these low levels of HS had no effect on P. ultimum inhibition by P. aeruginosa. The Fe, chelated by HS, was in part responsible for the decrease of P. ultimum inhibition by the bacteria when increasing amounts of HS were added in the culture medium. The addition of 500 mg l(-1) of humic acids isolated from de-inking paper sludge compost or from fossil origin completely eliminated the inhibition of P. ultimum by R. radiobacter. This Fe effect also stimulated growth of R. radiobacter and reduced its siderophore production in a minimal medium supplemented with HS as sole source of Fe. The results showed that HS influence microbial antagonism when added to a culture medium. However, this effect varies with different factors such as the type of bacteria, concentration of HS, molecular weight and Fe content.     

Composting of de-inking sludge from the recycled paper manufacturing industry

Composting of two different types of sludge from the recycled paper manufacturing industry was carried out at laboratory scale. Physico-chemical sludge (PCS) from the de-inking process and biological sludge (BS) from the wastewater treatment plant were composted and co-composted with and without addition of a bulking material. Despite its poor initial characteristics (relatively high C/N ratio, low organic content and moisture), PCS showed excellent behaviour in the composting process, reaching and maintaining thermophilic temperatures for more than 7 days at laboratory scale, and therefore complete hygienization. Pilot scale composting of PCS was also studied, and a respiratory quotient of 1.19 was obtained, indicating a full aerobic biological process. Respiration tests showed a complete stabilization of the material, with final values of the static respiration index in the range of 1.1 mg O2gTOM(-1)h(-1). Composting is proposed as a suitable technology for the effective recycling of this type of sludge from the recycled paper manufacturing industry.     

Dynamics of water-soluble carbon substances and microbial populations during the composting of de-inking paper sludge

Composting is an alternative method to dispose of de-inking paper sludge (DPS). Today, few studies have investigated the water-soluble carbon (WSC) substances as indicators of the decomposition process and the microbial changes taking place during the composting of DPS. Accordingly, the goal is to study their dynamics during the composting of DPS at three nitrogen levels, 0.6%, 0.7% or 0.9% total N, using mechanical turning. The changes in WSC substances, microbial biomass carbon (MBC) and, total and DPS microbial populations were monitored during 24 weeks. Also, microorganisms were identified and tested for the production of selected enzymes. Regardless of N treatments, the dynamic of WSC substances indicated that cellulose and hemicellulose fractions of DPS fibers were mainly biodegraded during the first 8 weeks while the more resistant carbon (C) fractions were biodegraded thereafter. MBC also evolved regardless of N treatments but was correlated to WSC substances. Its high values decline mostly after 12 weeks indicating the exhaustion of this source of C energy for microbial growth and the stabilisation of DPS organic matter. The dynamic and identified microorganisms were comparable to those observed in other composting processes. However, the results pointed out that those mostly implicated in the hydrolysis of DPS fibers were the thermophilic actinomycetes and fungi and, by comparison to the 0.6% or 0.7% N treatment, they decreased in presence of the 0.9% N treatment. Most microorganisms were hemicellulolytic bacteria, while actinomycetes and fungi were capable of degrading a wide variety of substrates. Overall, dynamics of WSC substances and microbial populations indicated that during composting, DPS decomposition obey a two phase decay while, contrary to the lowest N treatment, the 0.9% N treatment has slowed down this process by harming the important microbial populations implicated in the degradation of DPS fibers.     

Restoration of ecosystem function in an abandoned sandpit: plant and soil responses to paper de-inking sludge

1. In minesoil reclamation, the establishment of a sustainable plant cover requires the improvement of limiting conditions and the re-initiation of carbon (C) and nutrient cycling.
2. The approach used in this study for reclaiming an abandoned sandpit in Quebec, Canada, was based on a heavy organic amendment as an attempt to accelerate the reconstruction of a functional ecosystem.
3. The one-time intervention consisted of incorporating paper de-inking sludge into soil at two rates (0 and 105 dry t ha^–1), supplemented with nitrogen (N) at three rates (3, 6 and 9 g kg^–1 sludge) and phosphorus (P) at two rates (0·5 and 1·0 g kg^–1 sludge) followed by seeding (mid-summer) of Agropyron elongatum (Host) Beauv. (tall wheatgrass).
4. Standing biomass increased in the presence of sludge after both the first and second full growing seasons. High N application rates further increased yield, more importantly in the second season. The high P rate improved grass establishment in all cases. Ground cover increased with time and doubled in the presence of sludge whereas it declined in the absence of sludge. Phosphorus and N uptake was improved consistently in the presence of sludge.
5. Sludge application resulted in improved water retention and cation exchange capacities, and an increase in pH and bulk density of sandpit minesoil, all of which may have accounted for the significant improvement in plant responses. Levels of soil C and N suggest that this reconstructed system is approaching sustainability.
6. Adequate N and P supplements will accentuate the positive influence of sludge on revegetation.     

Isolation and characterization of thermophilic methanogenic bacteria from mushroom compost

Abstract During the first stage of the preparation of mushroom compost oxygen is believed to be readily available. However we measured methane in the evoking air above the compost piles and were able to isolate thermophilic methanogenic bacteria from this compost. The isolates grow only on H₂ and CO₂ as energy and carbon source and do not require complex factors for growth. On the basis of nutritional and morphological characteristics these methanogens were identified as strains of Methanobacterium thermoautotrophicum .     

Isolation of rhodoquinone-containing chemoorganotrophic bacteria from activated sludge

Abstract Several strains of Gram-negative aerobic chemoheterotrophic bacteria which produced rhodoquinone were isolated from activated sludge. All isolates contained a rhodoquinone with eight isoprene units as a major component in addition to the corresponding homologue of ubiquinone. The isolates seemed to make up a single taxon because of their homogeneity in phenotypic and chemotaxonomic properties, but could not be allocated to any known taxa of Gram-negative bacteria.     

Isolation and characterization of filamentous bacteria present in bulking activated sludge

Summary Several types of filamentous microorganisms were observed and identified in samples of poorly settling (bulking) activated sludge. The major types encountered and the frequency (percentage) of appearance in the total of all treatment plants sampled were: Eikelboom type 0041 (60), type 1701 (45), Haliscomenobacter hydrossis (35), type 021N (30), Thiothrix spp. (20), and Sphaerotilus natans (20). Isolation techniques and culture media were developed and used to recover 42 axenic strains of filamentous bacteria from sludge samples collected. The isolates were identified as strains of Thiothrix, Beggiatoa, S. natans, and Eikelboom types 021N, 1701, 0041, and 0803. Nutritional and differential characterization of the bacteria was important to the differentiation of groups which could not be easily distinguished on the basis of morphology. Although certain treatment plant operating parameters (organic loading) seemed associated with the presence of specific filamentous organism types, possible interaction among factors precluded definite establishment of a cause and effect relationship for most of the treatment plant characteristics and organisms observed.     

Diversity and activity of free-living nitrogen-fixing bacteria and total bacteria in organic and conventionally managed soils

Agricultural soils are heterogeneous environments in which conditions affecting microbial growth and diversity fluctuate widely in space and time. In this study, the molecular ecology of the total bacterial and free-living nitrogen-fixing communities in soils from the Nafferton Factorial Systems Comparison (NFSC) study in northeast England were examined. The field experiment was factorial in design, with organic versus conventional crop rotation, crop protection, and fertility management factors. Soils were sampled on three dates (March, June, and September) in 2007. Total RNA was extracted from all soil samples and reverse transcribed. Denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) were used to analyze nifH and 16S rRNA genes in order to study free-living diazotrophs and the total bacterial community, respectively. Crop rotation was shown to have a significant effect on total bacterial diversity (and that of free-living N fixers) (P ≤ 0.001). On all three dates, nifH activity was higher in the conventional crop rotation. In contrast, qPCR analysis of free-living N fixers indicated significantly higher levels of activity in conventionally fertilized plots in June (P = 0.0324) and in plots with organic crop protection in September (P = 0.0143). To our knowledge, the effects of organic and conventional farming systems on free-living diazotrophs have never been studied. An increased understanding of the impacts of management practices on free-living N fixers could allow modifications in soil management practices to optimize the activity of these organisms.     

Biodiversity of denitrifying and dinitrogen-fixing bacteria in an acid forest soil

Isolated soil DNA from an oak-hornbeam forest close to Cologne, Germany, was suitable for PCR amplification of gene segments coding for the 16S rRNA and nitrogenase reductase (NifH), nitrous oxide reductase (NosZ), cytochrome cd(1)-containing nitrite reductase (NirS), and Cu-containing nitrite reductase (NirK) of denitrification. For each gene segment, diverse PCR products were characterized by cloning and sequencing. None of the 16S rRNA gene sequences was identical to any deposited in the data banks, and therefore each of them belonged to a noncharacterized bacterium. In contrast, the analyzed clones of nifH gave only a few different sequences, which occurred many times, indicating a low level of species richness in the N2-fixing bacterial population in this soil. Identical nifH sequences were also detected in PCR amplification products of DNA of a soil approximately 600 km distant from the Cologne area. Whereas biodiversity was high in the case of nosZ, only a few different sequences were obtained with nirK. With respect to nirS, cloning and sequencing of the PCR products revealed that many false gene segments had been amplified with DNA from soil but not from cultured bacteria. With the 16S rRNA gene data, many sequences of uncultured bacteria belonging to the Acidobacterium phylum and actinomycetes showed up in the PCR products when isolated DNA was used as the template, whereas sequences obtained for nifH and for the denitrification genes were closely related to those of the proteobacteria. Although in such an experimental approach one has to cope with the enormous biodiversity in soils and only a few PCR products can be selected at random, the data suggest that denitrification and N2 fixation are not genetic traits of most of the uncultured bacteria.     

Effect of high compost temperature on enzymatic activity and species diversity of culturable bacteria in cattle manure compost

To clarify the characteristics of thermophilic bacteria in cattle manure compost, enzymatic activity and species diversity of cultivated bacteria were investigated at 54, 60, 63, 66 and 70 degrees C, which were dependent on composting temperature. The highest level of thermophilic bacterial activity was observed at 54 degrees C. Following an increase in temperature to 63 degrees C, a reduction in bacterial diversity was observed. At 66 degrees C, bacterial diversity increased again, and diverse bacteria including Thermus spp. and thermophilic Bacillus spp. appeared to adapt to the higher temperature. At 70 degrees C, bacterial activity measured as superoxide dismutase and catalase activity was significantly higher than at 66 degrees C. However, the decomposition rate of protein in the compost was lower than the rate at 66 degrees C due to the higher compost temperature.     

A microautoradiographic study of the activity of attached and free-living bacteria

Microautoradiography, combined with epifluorescent microscopy, was used to evaluate the uptake of tritiated amino acids by a marine Pseudomonas sp. A comparison was made between the activity of bacteria free-living in the medium and bacteria which were attached to glass, polyethylene or polystyrene substrata. The proportion of active bacteria was lower for free-living cells (53–82%) and those attached to polystyrene (53–76%) than for those attached to glass (77–99%) or polyethylene (73–96%). For bacteria attached to glass, assimilated labelled substrate was retained within the cell over 3 h, whereas with polyethylene, labelled material was released from the cells and adsorbed on the surrounding substratum. Hence the physiological activity of attached bacteria depended on the chemical composition of the substratum.     

Isolation of some new sulphur bacteria from activated sludge

E mtiazi , G., H abibi , M.H. & S etareh , M. 1990. Isolation of some new sulphur bacteria from activated sludge. Journal of Applied Bacteriology 69 , 864–870.
During studies on bulking of activated sludge some new sulphur micro-organisms, which were able to grow aerobically and anaerobically on reduced sulphur compounds, were isolated on thiosulphate agar. These were capable of autotrophic and heterotrophic growth on a wide range of substrates. In view of their ability to oxidize reduced sulphur compounds, and because one of them was an oxidase- and catalase-positive, Gram-negative, motile coccus (0.36–0.48 μm) it was named Thiosphaera persica . The second one was an oxidase- and catalase-positive, Gram-negative, motile rod (1.32–1.80 μm) and was named Thiobacillus persica . The third one was oxidase-negative, catalase-positive, motile, Gram-negative and polymorphic and was named Sulphobacter polymorpha .     

Composting of de-inking paper sludge with poultry manure at three nitrogen levels using mechanical turning: behavior of physico-chemical parameters.

De-inking paper sludge (DPS) is rich in carbon (C) but poor in nitrogen (N). Thus, it has a high C:N ratio which limits the composting process. Accordingly, the goal of this study was to investigate the effect of three N treatments on DPS composting. Compost piles of 100 m3 were formed by mixing raw DPS with poultry manure and chicken broiler floor litter, giving on average 0.6%, 0.7% and 0.9% total N. The changes in physico-chemical parameters, total weight and fiber losses, and maturity of composting piles of DPS were monitored during 24 weeks. The compost piles had a neutral to alkaline pH throughout the study. Inorganic N decreased whereas organic N increased over time for all treatments. These changes in magnitude were different among N treatments resulting in a final total N content of 0.9% for the 0.6% N treatment whereas final total N contents of 0.7% and 0.9% N were measured for the 0.7% and 0.9% N treatments. The total weight, cellulose and hemicellulose losses were higher in 0.6% N treatment giving the lowest C:N ratio after 24 weeks of composting. However, none of the 24 week-old composts of DPS were mature based on their final C:N ratio and colorimetric test of maturity. Except for copper, their final total trace element contents meet most known standards or guidelines for organic soil conditioners. Overall, 0.6% N treatment was the best to enhance DPS composting using mechanical turning, but a period of more than 24 weeks was required to reach compost maturity.     

具有ACC脱氨酶活性的细菌的分离和鉴定

在以ＡＣＣ（α－氨基环丙烷梭酸）为唯一氮源的选择性培养基上,对土壤来源的细菌菌株进行了筛选。通过生长测定、对ＡＣＣ降解的分析,确定了菌株ＡＣＣ脱氨酶的活性,并对菌株进行了系统鉴定。     

Isolation of hydrogen-producing bacteria from granular sludge of an upflow anaerobic sludge blanket reactor

H₂-producing bacteria were isolated from anaerobic granular sludge. Out of 72 colonies (36 grown under aerobic conditions and 36 under anaerobic conditions) arbitrarily chosen from the agar plate cultures of a suspended sludge, 34 colonies (15 under aerobic conditions and 19 under anaerobic conditions) produced H₂ under anaerobic conditions. Based on various biochemical tests and microscopic observations, they were classified into 13 groups and tentatively identified as follows: From aerobic isolates,Aeromonas spp. (7 strains),Pseudomonas spp. (3 strains), andVibrio spp. (5 strains); from anaerobic isolates,Actinomyces spp. (11 strains),Clostridium spp. (7 strains), andPorphyromonas sp. When glucose was used as the carbon substrate, all isolates showed a similar cell density and a H₂ production yield in the batch cultivations after 12h (2.24–2.74 OD at 600 nm and 1.02–1.22 mol H₂/mol glucose, respectively). The major fermentation by-products were ethanol and acetate for the aerobic isolates, and ethanol, acetate and propionate for the anaerobic isolates. This study demonstrated that several H₂ producers in an anaerobic granular sludge exist in large proportions and their performance in terms of H₂ production is quite similar.     

Isolation and characterization of two novel ethanol-tolerant facultative-anaerobic thermophilic bacteria strains from waste compost

In a search for potential ethanologens, waste compost was screened for ethanol-tolerant thermophilic microorganisms. Two thermophilic bacterial strains, M5EXG and M10EXG, with tolerance of 5 and 10% (v/v) ethanol, respectively, were isolated. Both isolates are facultative anaerobic, non-spore forming, non-motile, catalase-positive, oxidase-negative, Gram-negative rods that are capable of utilizing a range of carbon sources including arabinose, galactose, mannose, glucose and xylose and produce low amounts of ethanol, acetate and lactate. Growth of both isolates was observed in fully defined minimal media within the temperature range 50–80°C and pH 6.0–8.0. Phylogenetic analysis of the 16S rDNA sequences revealed that both isolates clustered with members of subgroup 5 of the genus Bacillus. G+C contents and DNA–DNA relatedness of M5EXG and M10EXG revealed that they are strains belonging to Geobacillus thermoglucosidasius. However, physiological and biochemical differences were evident when isolates M5EXG and M10EXG were compared with G. thermoglucosidasius type strain (DSM 2542^T). The new thermophilic, ethanol-tolerant strains of G. thermoglucosidasius may be candidates for ethanol production at elevated temperatures.     

Relative abundance of denitrifying and dinitrogen-fixing bacteria in layers of a forest soil

The populations of N(2)-fixing and denitrifying bacteria in an acid forest soil near Cologne were characterized by gene probing. The DNA isolated from the soil for this purpose was suitable for DNA-DNA hybridization using 0.4-0.7-kb probes targeting denitrification enzymes, dinitrogenase reductase (nifH) and eubacterial 16S rRNA. The densitometrical comparison of band intensities obtained in these Southern hybridizations indicated that the highest number of total bacteria, of denitrifying and N(2)-fixing microorganisms always occurred in the upper ( approximately 5 cm) soil layer. The concentration of all these organisms decreased in parallel with the soil depth. The soil investigated was rich in nitrate in all layers, and the availability of nitrate apparently did not govern the distribution of denitrifying and N(2)-fixing bacteria in this soil. Soil cores investigated in the laboratory formed N(2)O on addition of nitrate irrespective of the presence of C(2)H(2). Hybridization intensities, with a gene probe for the 16S rRNA, and MPN numbers were generally higher in soil samples taken from the roots of plants than in the bulk soil. There was no selective enrichment of denitrifying or N(2)-fixing bacteria at the roots. The data obtained by hybridizing isolated soil DNA generally matched previous results obtained with culturable bacteria.