Ammonia emissions during the initial phase of microbial degradation of solid and liquid cattle manure

The maximum specific ammonia emissions from liquid manure (LM) and solid manure containing 2.5 kg straw/livestock unit (LU)/day (SM 2.5) or 15 kg straw/LU/day (SM 15) increased in the sequence LM < SM 2.5 < SM 15 (662.6 < 3163.7 < 6299.8 μg NH₃–N/h/kg). These emission levels were attained soon after the maximum temperatures (22.9°C < 34.3°C < 69.5°C) induced by microbial self-heating had been reached. After that, NH₄⁺ was microbially re-bound in amounts that increased with a higher C content and a widening C:N ratio, i.e. also in the sequence LM < SM 2.5 < SM 15. Over a period of 15 to 16 days, 6.0% (LM), 10.8% (SM 2.5) and 5.9% (SM 15) of the N_total was emitted. When the accumulated ammonia emissions were extrapolated beyond this period of investigation, it was concluded that, over longer storage periods, solid manure offers better biological conditions for low ammonia emissions than liquid manure.     

Fixed-bed fermentation of rice straw and chicken manure using a mixed culture of marine mesophilic microorganisms

A mixture of rice straw (80%) and chicken manure (20%) was pretreated and fermented to carboxylic acids by using a mixed culture of marine mesophilic microorganisms. Two sets of four fermentors, built from PVC pipes, were used for both biomass pretreatment and fermentation. Four 1L fermentors (F1-F4) were arranged in series, where liquid fermentation products were transferred from one fermentor to the other, to form a train. A liquid volume of 10mL and 15mL were transferred every four days for Trains A and B, respectively. The maximum total acid concentration for F1 in Train A was 34.2g/L and the maximum acid concentration in F2-F4 was approximately 44g/L. The maximum total acid concentration in F1 in Train B was 30.5g/L and the maximum acid concentration in F2-F4 was approximately 48g/L. The conversion in each of the fermentors in Train A varied from 0.821 to 0.879g VS digested/g VS fed and the yield was in the range 0.489-0.609g total acids/g VS fed. The conversion and yield in Train B were 0.741-0.914g VS digested/g VS fed and 0.563-0.669g total acids/g VS fed, respectively. The continuum particle distribution model (CPDM) predicted acid concentrations and retention times in the fixed-bed fermentation system with R(2) of 0.67-0.84 in Trains A and B.     

Three-step biological process for the treatment of the liquid fraction of cattle manure

The liquid fraction of cattle manure was subjected to a biological treatment combining anoxic-anaerobic and oxic processes in order to stabilize the organic matter and reduce nitrogen and phosphorus so as to avoid problems of pollution when applying it to the land. The anoxic process was carried out at 30 degrees C in a CSTR reactor, the anaerobic process in a UASB reactor at 37 degrees C and the oxic treatment in another CSTR at 20 degrees C. The following results were obtained when working under optimum conditions (removal efficiencies in brackets): COD was reduced from 42 to 3.8 g/L (>90%); total solids from 41 to 14 (67%); total volatile solids from 22 to 7.0 (68%); total Kjeldahl nitrogen from 2.2 to 0.1 g/L (95%); NH4(-)-N from 1.10 to 0.02 g/L (98%) and Total-Phosphorus from 0.696 to 0.058 g/L (92%). Nitrates, undetected in the liquid fraction of cattle manure, were present in the final effluent as a result of nitrification. To reduce the amount of nitrates, different recirculation rates were tested. The minimum nitrate concentration achieved was 127 mg/L using a recirculation ratio of 4.     

Inhibition of methanogenesis and its reversal during biogas formation from cattle manure

The composition of volatile fatty acids in the biogas digester based on cattle manure as substrate and stabilised at 25°C showed that it contained 87–88% branched chain fatty acids, comprising of isobutyric and isovaleric acids, in comparison to 38 % observed in the digester operating at 35°C. Mixed cellulolytic cultures equilibrated at 25°C (C-25) and 35‡C (C-35) showed similar properties, but rates of hydrolysis were three times higher than that observed in a standard biogas digester. The proportion of isobutyric and isovaleric were drastically reduced when C-25 was grown with glucose or filter paper as substrates. The volatile fatty acids recovered from C-25 (at 25°C) inhibited growth of methanogens on acetate, whereas that from C-35 was not inhibitory. The inhibitory effects were due to the branched chain fatty acids and were observed with isobutyric acid at concentrations as low as 50 ppm. Addition of another micro-organismRhodotorula selected for growth on isobutyric completely reversed this inhibition. Results indicate that the aceticlastic methanogens are very sensitive to inhibition by branched chain fatty acids and reduction in methane formation in biogas digester at lower temperature may be due to this effect.     

Effects of farmyard manure and phosphorus on zinc transformations and phyto-availability in two alfisols of India

Laboratory pot experiments were run to study the effects of added zinc (Zn) with and without farmyard manure (FYM) and phosphorus (P) on Zn transformations in two Alfisols, together with Zn uptake by wheat plants grown up to 60 days. In the first experiment the treatments included four levels of Zn (0, 3.75, 7.5 and 15 mgkg(-1) soil) and two levels of FYM (0 and 10 tha(-1)), and in the second experiment five levels of P (0, 20, 40, 80 and 160 mgkg(-1) soil) and one level of Zn (7.5 mgkg(-1) soil). The soils were sequentially fractionated into water-soluble plus exchangeable (CA-Zn), inorganically bound (AAC-Zn), organically bound (PYR-Zn), oxide bound (OX-Zn) and residual (RES-Zn) forms. The effect of added FYM was more evident on the OX-Zn fraction and the percentage utilization of Zn by wheat was the greatest with the addition of FYM alone at the rate of 10 tha(-1) (1.95-2.38%) in comparison to other treatment combinations. Among the levels, application of 7.5 mg Zn kg(-1) soil showed the maximum increase in different fractions of soil Zn and significantly increased the Zn utilization by wheat (0.87-1.17%) as compared to other Zn levels (0.58-0.88%). On an average, about 85% of the added Zn was recovered in different fractions in Zn treated pots. However, the recovery per cent of the added Zn was significantly higher at Zn level 7.5 (95%) mgkg(-1) soil than at 3.75 (87%) and 15 (73%) mg Zn kg(-1) soil levels. Phosphorus additions up to 40 mgkg(-1) soil increased the plant-available Zn in soils whereas at higher P levels plant-available forms decreased with a concominant increase in the inert forms. At 160 mg P kg(-1) soil, the P effect was more pronounced in the shoot than in the root, suggesting that a higher P level inhibits Zn translocation from root to upper plant parts. Path analysis showed that the organically (PYR-Zn) and inorganically bound (AAC-Zn) Zn fractions were the predominant fractions that influenced the Zn availability to plants.     

Factors controlling nitrification and denitrification: A laboratory study with gel-stabilized liquid cattle manure

Nitrification and denitrification were studied in a millimeterscale microenvironment using a two-phase system with a liquid manure-saturated layer. Samples consisted of liquid cattle manure and air-dried soil stabilized with silica gel, placed between two aerobic soil phases with a water content near field capacity. A high potential for NH₄ ⁺ oxidation developed within 0–2 mm distance from the interface, and NH₄ ⁺ diffused only 10–20 mm into the soil. Some NH₄ ⁺ was probably immobilized by microorganisms in the soil between 0 and 4 days, after which nitrification was the only sink for NH₄ ⁺. A potential for denitrification developed within the manure-saturated zone. Maximum rates of both potential and actual denitrification were recorded by Day 4, but denitrification continued for at least 2–3 weeks. The potential for nitrification peaked after 14 days. When the pH of the manure was adjusted to 5.5, nitrification was reduced close to the interface, and NH₄ ⁺ penetrated further into the soil before it was oxidized. The pH adjustment had an inhibitory effect on denitrification: Both potential and actual rates of denitrification were almost eliminated for several days. The size of the manure-saturated layer strongly affected denitrification losses. With layers of 8 and 16 mm thickness, losses equivalent to 33 and 40% of the original NH₄ ⁺ pool, respectively, were estimated. When manure corresponding to a 12 mm layer was homogeneously mixed with the soil, only 0.3% was lost.Offprint requests to: S. O. Petersen.     

Effect of microorganisms on rate of liquid extraction of ethanol from fermentation broths

Liquid extraction is one means of removing metabolic products continuously during a fermentation and so reducing product inhibition. It is known that microbial organisms are attracted to liquid-liquid interfaces, and it is important for the design of extraction systems to establish if this has a detrimental effect on the rate of extraction. The extraction of ethanol from aqueous suspensions of yeast (Saccharomyces cerevisiae) using n- decanol is described in this paper. It was found that the presence of the yeast cells severely reduced the rate of ethanol extraction. The overall mass transfer coefficient was reduced from 5.0 x 10(-6) to 0.7 x 10(-6) m/s. This reduced overall mass transfer coefficient was unaffected by yeast concentration in the range 0.1-20 kg/m(3). The results are consistent with the yeast cells adsorbing to the interface in closely packed layers and preventing mass transfer by simply reducing the available interfacial area. Optical microscope observations confirmed that a yeast layer several cell diameters thick rapidly built up at the interface when a small decanol droplet was added to a yeast suspension.     

Decomposition of nucleic acids and some of their degradation products by microorganisms

A study was made of the decomposition of nucleic acids, uric acid and urea by different groups of soil microorganisms including bacilli, non-coryneform rods, corynebacteria (arthrobacters and non-arthrobacters), streptomycetes, fungi and yeasts. Hydrolysis of nucleic acids was found to be a common phenomenon. The decomposition of uric acid was readily carried out by arthrobacters and streptomycetes. Most bacilli, however, lacked this capacity. Few soil microorganisms degraded urea. Additional investigations were made of the breakdown of nucleic acids and their degradation products by coryneform bacteria from soil, cheese, sea-fish, sewage, the phyllosphere and poultry litter. Generally, coryneforms from soil could utilize allantoin as the sole source of nitrogen, carbon and energy in contrast to most of those from cheese which could not. Breakdown of uric acid and allantoin by washed cells of a coryneform strain from soil resulted in formation of ammonia; breakdown of these compounds by washed cells of a strain from cheese resulted in accumulation of urea.     

Prevalence of verotoxin-producing Escherichia coli (VTEC) in slurry,farmyard manure and sewage sludge in France

AIMS: The aims of the present study were to determine VTEC prevalence in manure, slurry and sewage sludge in France and to characterize the VTEC strains isolated (virulence genes and serotype). METHODS AND RESULTS: Seven hundred and fifty-two samples from 55 farmyard manures, 136 bovine and porcine faeces, 114 slurries, 10 composts, and 437 samples from outflows of sewage wastewater treatment plants were analysed. Twenty-four percent contained isolates which were PCR positive for stx gene. Twenty-one VTEC strains were recovered from positive samples by colony hybridization: 76% of them were positive for stx(2) gene, 33% for stx(1) gene,and 19% for eae gene. One strain belonged to serotype O157:H7 and two others to serogroups O26 and O55, respectively. CONCLUSIONS: Some of the VTEC strains isolated from environments in France should be considered as potentially pathogenic for humans. SIGNIFICANCE AND IMPACT OF THE STUDY: Appropriate handling or use of manure, slurry and sewage sludge is necessary so that contamination of the environment and food by VTEC can be prevented.     

High-pressure liquid chromatography of polycyclic aromatic hydrocarbons and some of their derivatives.

The separation of polycyclic aromatic hydrocarbons and their derivatives by means of high-pressure liquid chromatography on Permaphase ODS is described. The method consists of the (isocratic) elution of compounds from the column with a methanol-water mixture of constant composition and is particularly suited to the identification of metabolic products of polycyclic hydrocarbons.     

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 influence of some herbicides on selected groups of microorganisms in loess and sandy soils. II. The effect of nitrification.

Studies on the effect of herbicides on nitrification in field and laboratory conditions have shown that steady use of these substances in the same field does not exert a significant effect on this process in the soil. The herbicides caused only shortlasting disturbances. Nitrification usually returned to a relative equilibrium in several days after the herbicide application.     

Uncoupling proteins in mitochondria of plants and some microorganisms.

Uncoupling proteins, members of the mitochondrial carrier family, are present in mitochondrial inner membrane and mediate free fatty acid-activated, purine-nucleotide-inhibited H+ re-uptake. Since 1995, it has been shown that the uncoupling protein is present in many higher plants and some microorganisms like non-photosynthetic amoeboid protozoon, Acanthamoeba castellanii and non-fermentative yeast Candida parapsilosis. In mitochondria of these organisms, uncoupling protein activity is revealed not only by stimulation of state 4 respiration by free fatty acids accompanied by decrease in membrane potential (these effects being partially released by ATP and GTP) but mainly by lowering ADP/O ratio during state 3 respiration. Plant and microorganism uncoupling proteins are able to divert very efficiently energy from oxidative phosphorylation, competing for deltamicroH+ with ATP synthase. Functional connection and physiological role of uncoupling protein and alternative oxidase, two main energy-dissipating systems in plant-type mitochondria, are discussed.     

Successive rapid reductive dehalogenation and mineralization of pentachlorophenol by the indigenous microflora of farmyard manure compost

AIMS: To determine whether composting with animal manure can be used to effectively remediate soil from a pentachlorophenol (PCP)-contaminated site, and to establish the fate of the degraded xenobiotic. METHODS AND RESULTS: Contaminated soil from a sawmill site was mixed with farm animal manure and composted in a 0.5 m3 silo under fully aerobic conditions. The disappearance and fate of PCP was monitored by gas chromatography (GC-ECD) and extensive mineralization confirmed in experiments with 14C-radiolabelled PCP. The disappearance of PCP was rapid and virtually complete within 6 days, prior to the onset of thermophilic conditions. Dechlorination of the PCP was found to be both reductive and sequential. CONCLUSIONS: PCP removal from contaminated soil by aerobic composting with animal manure is efficient and proceeds via reductive dechlorination to virtually complete mineralization. This contrasts with other chlorophenol composting regimes in which mineralization is achieved but dechlorination intermediates do not accumulate to detectable levels. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study demonstrate that anaerobic reductive dechlorination can proceed in an aerobic composting environment and contribute to efficient pentachlorophenol removal. Farmyard manure composts may represent a rapid, low-cost, low-technology option for treatment of chlorophenol-contaminated soils.