The impact of sludge amendment on gas dynamics in an upland soil: monitored by membrane inlet mass spectrometry

Studies of the land disposal of biosolids and municipal sewage have focused largely on the potential pollution of the soil with pathogens, toxic compounds or heavy metals. Little is known about the impact of sludge amendment on carbon source and sink concentrations in soils. In this study gas concentrations in Scottish soil cores (from limed and unlimed plots) were monitored continuously at 3 cm depth before, during and after sludge application using membrane inlet mass spectrometry (MIMS). Following sludge application to soil cores, significant and sustained increases in CH4 (for 8 days) and CO2 (for between 16 and 120 days) concentration were observed. This suggested short-term stimulation of indigenous methanogens, provision of a new methanogenic inoculum, or inhibition of methane oxidizers (for example by heavy metals or NH4 in sludge). Soil microbial fermentative activity was enhanced over periods of a few months as shown by elevated CO2 concentrations.     

Novel approach to control Salmonella enterica by modern biophotonic technology: photosensitization

Aims:  Salmonellosis is one of the most common foodborne diseases in the world. The aim of this study was to evaluate the antibacterial efficiency of 5-aminolevulinic acid (ALA) based photosensitization against one of food pathogens Salmonella enterica .
Methods and Results:  Salmonella enterica was incubated with ALA (7·5 mmol l⁻¹) for 1–4 h and afterwards illuminated with visible light. The light source used for illumination of S. enterica emitted light λ  = 400 nm with energy density 20 mW cm⁻². The illumination time varied from 0 to 20 min and subsequently a total energy dose reached 0–24 J cm⁻². The data obtained indicate that S. enterica is able to produce endogenous photosensitizer PpIX when incubated with ALA. Remarkable inactivation of micro-organisms can be achieved (6 log) after photosensitization. It is obvious that photosensitization-based inactivation of S. enterica depends on illumination as well as incubation with ALA time.
Conclusion:  ALA-based photosensitization can be an effective tool against multi-drug resistant Gram-negative bacteria S. enterica serovar Typhimurium.
Significance and Impact of the Study:  Experimental data and mathematical evaluations support the idea that ALA-based photosensitization can be a useful tool for the development of nonthermal food preservation technology in future.     

Listeria monocytogenes contamination of crops grown on soil treated with sewage sludge cake

A l -G hazali , M.R. & A l -A zawi , S.K. 1990. Listeria monocytogenes contamination of crops grown on soil treated with sewage sludge cake. Journal of Applied Bacteriology 69 , 642–647.
Listeria monocytogenes was found in the sewage sludge cake which is commonly used as an agricultural fertilizer in Iraq. Soils treated with this material were contaminated with the organism. Pot and field experiment showed that crops grown on treated soil became contaminated with L. monocytogenes and when alfalfa plant was grown on farmland soil treated with sewage sludge cake, listerias were found on 10% of 50 plants sampled at harvest, but the organism was detected only in low numbers on these crops (≤ 5 cells/g). This could add to the risk to animals and man.     

Contamination Levels Remaining in Aznalcóllar Spill-Affected Soils (Spain) Following Pyritic Sludge Removal

Potentially toxic elements (PTE) in aqua regia extracts were analyzed by ICP in order to determine the remaining degree of contamination of the selected points along Agrio and Guadiamar rivers. The studied area was affected in 1998 by the accidental spillage from a flotation plant for complex sulfides located at Aznalcóllar (Spain), it was covered by acidic waters and pyritic sludge. Emergency response for the remediation of the contaminated soils, for avoiding greater ecological impact, and waiting for possible ulterior remedial actions, was the removal of the superficial sludge. Sampling was done after the sludge removal operations 1 year after the spill, at different depths down to 300?cm. The results show that the soils affected are still contaminated by several PTE at different degrees and depths depending on their particular conditions. In general, superficial soil samples became highly contaminated. The contamination of soils is due, in general, to processes of sludge infiltration and PTE leaching. The sludge infiltration is due, in part, to the heavy machinery used to remove the sludge. A moderate degree of contamination remains in some soils down to 120 or 300?cm.     

A new set of PCR assays for the identification of multiple human adenovirus species in environmental samples

Aims:  To assess human adenovirus (HAdV) diversity in environmental samples based on sequence comparisons of hexon gene fragments amplified using newly designed HAdV-specific polymerase chain reaction (PCR) assays.
Methods and Results:  Six PCR primer sets were designed based on 56 aligned hexon sequences from NCBI GenBank to amplify different hexon gene sections (241–349 bp) of the six HAdV species. The amplified hexon genes from wastewater samples were cloned, sequenced, and compared with those in publicly accessible databases (i.e. NCBI GenBank) by using the B last program. A total of 46 analysed positive clones were affiliated to five HAdV serotypes, i.e. 1, 2, 12, 31 and 41. Similarities between the cloned and database hexon sequences ranged from 95·9 to 100% (with an average of 98·1 ± 1·0%).
Conclusion:  The designed primers showed higher amplification efficiencies when compared with the existing assays. Using the new assays, HAdV species A, C, and F (serotypes 1, 2, 12, 31 and 41 in particular) were identified in the studied municipal wastewater.
Significance and Impact of the Study:  The six PCR primer sets developed in this study can be used to efficiently amplify hexon gene fragments in HAdV. Multiple HAdV serotypes identified in the municipal wastewater provide new information about HAdV diversity in environmental samples.     

Mobility and Distribution of Zinc,Cadmium and Lead in Calcareous Soils Receiving Spiked Sewage Sludge

Leaching column experiments were conducted to determine the degree of mobility and the distribution of zinc (Zn), cadmium (Cd), and lead (Pb) because of an application of spiked sewage sludge in calcareous soils. A total of 20 leaching columns were set up for four calcareous soils. Each column was leached with one of these inflows: sewage sludge (only for two soils), spiked sewage sludge, or artificial well water (control). The columns were irrigated with spiked sewage sludge containing 8.5 mg Zn l^?1, 8.5 mg Cd l^?1, and 170 mg Pb l^?1 and then allowed to equilibrate for 30 days. At the end of leaching experiments, soil samples from each column were divided into 18 layers, each being 1 cm down to 6 cm and 2 cm below that, and analyzed for total and extractable Zn, Cd and Pb. The fractionation of the heavy metals in the top three layers of the surface soil samples was investigated by the sequential extraction method. Spiked sewage sludge had little effect on metal mobility. In all soils, the surface soil layers (0-1 cm) of the columns receiving spiked sewage sludge had significantly higher concentrations of total Zn, Cd and Pb than control soils. Concentration of the heavy metals declined significantly with depth. The mobility of Zn was usually greater than Cd and Pb. The proportion of exchangeable heavy metals in soils receiving spiked sewage sludge was significantly higher than that found in the control columns. Sequential extraction results showed that in native soils the major proportion of Zn and Pb was associated with residual (RES) and organic matter (OM) fractions and major proportion of Cd was associated with carbonate (CARB) fraction, whereas after leaching with spiked sewage sludge, the major proportion of Zn and Pb was associated with Fe-oxcide (FEO), RES, and CARB fractions and major proportion of Cd was associated with CARB, RES and exchangeable (EXCH) fractions. Based on relative percent, Cd in the EXCH fraction was higher than Zn and Pb in soils leached with spiked sewage sludge.     

Effects of soil amendment on gas depth profiles in soil monoliths using direct mass spectrometric measurement

Land use and agricultural practices are known to influence the source and sink concentrations of various gases, including greenhouse gases (NOx CH4 and CO2). in soils. With everincreasing production of domestic sewage sludge and the prohibition of disposal at sea, pressure on waste disposal increases. Anaerobically digested domestic sewage sludge and/or lime were applied to an upland. Scottish soil and their effects on gas depth profiles monitored as indicators of microbial processes of the soil ecosystem. The concentrations of various gases (Ar, O2. CO2, CH4, N2, NOx) were measured simultaneously at each depth using membrane inlet mass spectrometry (MIMS). This technique enables the direct measurement of multiple gas species throughout soil cores with minimal disturbance. Intact soil monoliths were collected from the sample site, following amendment, and maintained in a constant temperature, environmental growth chambers. Statistical analyses (one-way ANOVA and LSD tests) were conducted to identify the depths at which gas concentrations in amended cores were significantly different from those in control (un-amended) cores. Significant effects were observed on the concentration of CO2, CH4, NOx and N2 at certain depths. Average CH4 concentration was consistently higher (>1 microM) in the upper horizon following application of sludge and sludge and lime together. N2 and NOx concentrations were elevated in cores treated with lime by approximately 100 and 32 microM. respectively, in much of the upper horizon. CO2 concentration increased above control mean values, at certain depths, following application of either sludge or lime. Some explanation for the changes in soil gas concentration was provided by reference to the microorganism assemblages and the gases associated with biochemistry of nitrification, denitrification, methane oxidation and methanogenesis.     

Poliovirus retention in 75-cm soil cores after sewage and rainwater application.

The adsorption rate of a guanidine-resistant strain of poliovirus LSc 2ab was measured in Long Island soils with in situ field cores (10.1 by 75 cm). The test virus was chosen because it exhibited soil adsorption and elution characteristics of a number of non-polioviruses. After the inoculation of cores with seeded sewage effluent at a 1-cm/h infiltration rate, cores were extracted, fractionated, and analyzed for total plaque-forming units per each 5-cm fraction. The results showed that 77% of the viruses were adsorbed in the first 5 cm of soil. An additional 11% were found in the 5- to 10-cm fraction, and a total of 96% of the viruses were adsorbed by 25 cm. The remaining 4% were uniformly distributed over the next 50 cm of soil, with a minimum of 0.23% in each soil section. Few viruses (< 0.22%) were observed in core filtrates. Analysis of the viral distribution pattern in seeded cores, after an application of a single rinse of either sewage effluent or rainwater, indicated that large-scale viral mobilization was absent. However, localized areas of viral movement were noted in both of the rinsed cores, with the rainwater-rinsed cores exhibiting more expensive movement. All mobilized viruses were resorbed at lower core depths.     

Quantification and stability of human adenoviruses and polyomavirus JCPyV in wastewater matrices

Human adenoviruses (HAdV) and human polyomavirus JCPyV have been previously proposed as indicators of fecal viral contamination in the environment. Different wastewater matrices have been analyzed by applying real-time quantitative PCR procedures for the presence, quantity, and stability of a wide diversity of excreted HAdV and JCPyV. High quantities of HAdV and JCPyV were detected in sewage, effluent wastewater, sludge, and biosolid samples. Both viruses showed high stability in urban sewage. These results confirm the suitability of both viruses as indicators of human fecal viral pollution.     

Quantification and Stability of Human Adenoviruses and Polyomavirus JCPyV in Wastewater Matrices

Human adenoviruses (HAdV) and human polyomavirus JCPyV have been previously proposed as indicators of fecal viral contamination in the environment. Different wastewater matrices have been analyzed by applying real-time quantitative PCR procedures for the presence, quantity, and stability of a wide diversity of excreted HAdV and JCPyV. High quantities of HAdV and JCPyV were detected in sewage, effluent wastewater, sludge, and biosolid samples. Both viruses showed high stability in urban sewage. These results confirm the suitability of both viruses as indicators of human fecal viral pollution.     

Poliovirus retention in 75-cm soil cores after sewage and rainwater application

The adsorption rate of a guanidine-resistant strain of poliovirus LSc 2ab was measured in Long Island soils with in situ field cores (10.1 by 75 cm). The test virus was chosen because it exhibited soil adsorption and elution characteristics of a number of non-polioviruses. After the inoculation of cores with seeded sewage effluent at a 1-cm/h infiltration rate, cores were extracted, fractionated, and analyzed for total plaque-forming units per each 5-cm fraction. The results showed that 77% of the viruses were adsorbed in the first 5 cm of soil. An additional 11% were found in the 5- to 10-cm fraction, and a total of 96% of the viruses were adsorbed by 25 cm. The remaining 4% were uniformly distributed over the next 50 cm of soil, with a minimum of 0.23% in each soil section. Few viruses (< 0.22%) were observed in core filtrates. Analysis of the viral distribution pattern in seeded cores, after an application of a single rinse of either sewage effluent or rainwater, indicated that large-scale viral mobilization was absent. However, localized areas of viral movement were noted in both of the rinsed cores, with the rainwater-rinsed cores exhibiting more expensive movement. All mobilized viruses were resorbed at lower core depths.     

Distribution and genotypic characterization of Salmonella serovars isolated from tropical seafood of Cochin, India

Aims:  To determine the distribution of Salmonella serovars in seafood and to examine the intraserovar genetic variations in Salmonella enterica subsp. enterica serovar Rissen and Salmonella Weltevreden by polymerase chain reaction (PCR)-ribotyping and enterobacterial repetitive intergenic consensus (ERIC)-PCR methods.
Method and Results:  A total of 417 seafood samples collected over 2003–2006 from fishing harbours and fish markets of Cochin (India) was studied for presence of Salmonella serovars. Seafood samples were analysed for the presence of Salmonella by Bacteriological Analytical Manual (BAM), U.S. Food & Drug Administration (USFDA) method. The study indicated that 23·2% of the seafood samples were positive for Salmonella and a total of 241 Salmonella isolates comprising of 27 different serovars were isolated from seafood. S. Weltevreden, Salmonella Rissen, Salmonella Typhimurium and Salmonella Derby were found to be the most predominant serovars in seafood. PCR-ribotypes and ERIC-PCR profiles showed multiple genotypic profiles for S. Rissen and S. Weltevreden in seafood and the level of discrimination indices obtained was at 0·974 for S. Rissen and 0·988 for S. Weltevreden, respectively.
Conclusion:  The study highlighted the major Salmonella serovars in the seafood of Cochin (India) and molecular fingerprinting pattern revealed genetic variation among S. Rissen and S. Weltevreden.
Significance and Impact of the Study:  Widespread occurrence of Salmonella contamination in seafood and multiple clones of S. Rissen and S. Weltevreden detected in seafood, thus, indicated the diverse routes of Salmonella contamination in seafood.     

Application of microbial source tracking methods in a Gulf of Mexico field setting

Aims:  Microbial water quality and possible human sources of faecal pollution were assessed in a Florida estuary that serves shellfishing and recreational activities.
Methods and Results:  Indicator organisms (IO), including faecal coliforms, Escherichia coli and enterococci, were quantified from marine and river waters, sediments and oysters. Florida recreational water standards were infrequently exceeded (6–10% of samples); however, shellfishing standards were more frequently exceeded (28%). IO concentrations in oysters and overlaying waters were significantly correlated, but oyster and sediment IO concentrations were uncorrelated. The human-associated esp gene of Enterococcus faecium was detected in marine and fresh waters at sites with suspected human sewage contamination. Lagrangian drifters, used to determine the pathways of bacterial transport and deposition, suggested that sediment deposition from the Ochlockonee River contributes to frequent detection of esp at a Gulf of Mexico beach.
Conclusions:  These data indicate that human faecal pollution affects water quality in Wakulla County and that local topography and hydrology play a role in bacterial transport and deposition.
Significance and Impact of the Study:  A combination of IO enumeration, microbial source tracking methods and regional hydrological study can reliably inform regulatory agencies of IO sources, improving risk assessment and pollution mitigation in impaired waters.     

Evaluation of PCR assays for quantifying seed-borne infection by Fusarium and Microdochium seedling blight pathogens

Aims:  To evaluate competitive PCR assays for quantifying seed-borne Microdochium and Fusarium seedling blight pathogen DNA and to determine test and year repeatability and sources of variability.
Methods and Results:  Relationships between DNA and plate counts were significant for Fusarium and Microdochium seedling blight pathogens in 152 seed batches from 3 years. Coefficient of determinations, however, differed greatly ( Fusarium ; R ² = 0·25, P  =   0·029, Microdochium ; R ² = 0·73, P  <   0·001). Significant differences between years were observed in the regression slopes for Microdochium . Pathogen DNA quantified in 16 extractions after sampling was highly correlated to results following storage for 1–2 years ( R ² > 0·90). Residual maximum likelihood analysis showed that the least and greatest variance components of the testing procedure were DNA extraction subsampling and PCR assay respectively.
Conclusions:  Amount of pathogen DNA is a useful estimator of seed batch contamination for Microdochium but not Fusarium seedling blight pathogens. Although reproducible over time, improvements to the testing procedure should focus on repeated PCR amplifications to reduce assay variability.
Significance and Impact of the Study:  Replacing plate counts with competitive PCR for determining the severity of seed batch contamination is feasible in areas where Microdochium seedling blight pathogens predominate.     

Uptake and persistence of human associated Enterococcus in the mussel Mytilus edulis: relevance for faecal pollution source tracking

Aims:  Micro-organisms and molecular markers for microbial source tracking (MST) in coastal waters are often present at low numbers, and often exhibit significant variability in time and space. In this study, we investigated the uptake, accumulation, and persistence of human associated Enterococcus in the mussel Mytilus edulis .
Methods and Results:  The human associated molecular markers esp in Enterococcus faecium , and M66 in Enterococcus faecalis were targetted by PCR in seawater and mussel samples from coastal sites affected by sewage contamination. Both native mussels and mussels transplanted from pristine to polluted sites were included. The results showed that the esp and M66 markers were often not detectable in seawater whereas mussels were enriched in the markers. Human associated E. faecalis accumulated rapidly in M. edulis , and reached maximum levels after 4–6 h with concentration 30–300 times greater than in the surrounding seawater. Enterococcus faecalis retained in M. edulis showed a survival comparable to planktonic E. faecalis in seawater with half lives of 30 and 22 h, respectively. Human associated markers remained detectable for 120 h in M. edulis after faecal contamination.
Conclusions:  The study demonstrated that native and transplanted M. edulis can accumulate and retain human associated molecular markers relevant for MST.
Significance and Impact of the Study:  Mussels should be considered as additional targets in MST studies in coastal waters.     

25S rDNA-based molecular monitoring of glomalean fungi in sewage sludge-treated field plots

Recycling of sewage wastes in agriculture is likely to affect the biological activity of soils through contamination of ecosystems by pathogens and metallic or organic micropollutants. The impact of sewage sludge spreading under field conditions on arbuscular mycorrhiza (AM) formation by a community of glomalean fungi was evaluated using a nested polymerase chain reaction (PCR) and discriminating primers based on 25S rDNA polymorphisms to detect different fungal species within root systems. Medicago truncatula was grown in soil of field plots amended or not with a composted sewage sludge, spiked or not with organic or metallic micropollutants. Overall AM development in roots decreased with sewage sludge application, and the relative abundance of five AM fungal morphotypes in root fragments was modified by the input of composted sludges. Sewage sludge spiked or not with organic pollutants had a generally positive effect on the relative diversity of AM fungal populations in planta , whereas after spreading of the sludge spiked with metallic pollutants, no variation was observed in the abundance of different species.     

Application of luminescent biosensors for monitoring the degradation and toxicity of BTEX compounds in soils

Aims:  To assess the changes in acute toxicity and biodegradation of benzene, toluene, ethylbenzene and xylene (collectively referred to as BTEX) compounds in soil over time and compare the performances of biological and chemical techniques.
Methods and Results:  Biological methods ( lux -based bacterial biosensors, basal respiration and dehydrogenase activity) were related to changes in the concentration of the target compounds. There was an initial increase in toxicity determined by the constitutively expressed biosensor, followed by a continual reduction as degradation proceeded. The biosensor with the BTEX-specific promoter was most induced when BTEX concentrations were highest. The treatment with nutrient amendment had a significant increase in microbial activity, while the sterile control produced the lowest level of degradation.
Significance and Impact of the Study:  Luminescent biosensors were able to monitor changes in contaminant toxicity and bioavailability in aqueous extracts from BTEX-impacted soils as degradation proceeded. The integration of biological tests with chemical analysis enables a fuller understanding of the biodegradation processes occurring at their relative rates.
Conclusions:  The biological methods were successfully used in assessing the performance of different treatments for enhancing natural attenuation of BTEX from contaminated soils. While, chemical analysis showed biodegradation of parent BTEX compounds in biologically active soils, the biosensor assays reported on changes in bioavailability and potentially toxic intermediate fractions as they estimated the integrative effect of contaminants.     

Impact of application of organic waste materials on microbial and enzyme activities of mine soils in the Lusatian coal mining region

The objective of the present work was to study the short-term stimulation of microbial and enzyme activity in mine soils by application of organic waste materials in lysimeter and mesocosm studies. The mine soils derived from tertiary and quaternary deposits were ameliorated with brown coal filter ash (tertiary deposits) and lime (quaternary deposits). At the beginning of recultivation the soils were treated with varying amounts of sewage sludge, coal sludge, composted sewage sludge and compost to a depth of 30 cm. In the first 2 years after application of organic waste materials we found a very low level of microbial properties especially in the sandy materials from quaternary deposits but a significant increase in microbial respiration, substrate induced respiration and enzyme activities like invertase and alkaline phosphatase with increasing application rates of sewage sludge, compost and sewage sludge mixed with coal sludge. This can be explained by an increase in organic matter and nutrient content of the soils and an improvement of soil physical properties such as water and nutrient retention capacity. Additionally it can be assumed, that constituents of the coal admixtures of tertiary deposits can be mineralised or converted by the soil microorganisms. In the tertiary materials ameliorated with brown coal ash the highest amounts of microbial and enzyme activities were measured after application of nitrogen-rich sewage sludge or very high amounts of mature compost mainly consisting of green waste. Compared with sewage sludge the stimulating effects of composted sewage sludge were quite lower because of organic matter fragmentation and a reduced energy and nutrient supply to soil microorganisms. This revised version was published online in June 2006 with corrections to the Cover Date.     

Using Maize (Zea mays L.) and Sewage Sludge to Remediate a Petroleum-Contaminated Calcareous Soil

Phyto-stimulation, the use of plants to stimulate activity of microorganisms in a root zone, has been proposed as an approach to promote the degradation of petroleum hydrocarbons and thus the remediation of petroleum-polluted soils. In this study, we investigated the potential use of sewage sludge to enhance phyto-stimulating effects of maize (Zea mays L.) on the elimination of an aged petroleum contamination in a calcareous soil. In a pot experiment, maize was grown on the experimental soil for two months at three levels of sewage sludge application (0, 20, and 50 g dry matter of sludge per kg soil). The amendments increased root and shoot growth of the experimental plants approximately by a factor of two at the lower sludge treatment level and by a factor of five at the higher sludge treatment level. In a separate incubation experiment, sludge application also led to an immediate stimulation of soil respiration, which then further increased over time. The initial stimulation was three times larger at the higher than at the lower treatment level, but the rate of subsequent increase was similar in both treatments. The two sludge treatments also accelerated TPH elimination in the contaminated soil, and again the effect was approximately three times stronger at the higher than at the lower treatment level. The sludge effect on TPH elimination was much stronger than the effect of the plants. More than half of the initial contamination was reduced in combined treatment with maize and sludge application at the highest rate. The results show that sewage sludge can substantially enhance the remediation of petroleum-contaminated soil, especially when applied in conjunction with a suitable plant such as maize.     

Utilization of a sewage sludge for rehabilitating the soils degraded by the metallurgical industry and a possible environmental risk involved

Soils collected in the proximity of a steel plant in Krakow were analyzed for their pH, the content of total organic carbon, and the total contents of metals and metalloids. Applying the Commission of European Communities Bureau of Reference (BCR) extraction of selected metals: Cd, Fe, Pb, and Zn, their forms of binding in soils have been determined. The results of the soil analyses of the samples taken in 1996 and 2016 were compared, and then the soil contamination changes were assessed using the GI, contamination factor (CF), and PLI indicators. A rehabilitation of the degraded soils was tested using the communal sewage sludge, which was added into the soil at four various ratios. The sewage sludge applied meets the environmental standards as a material for rehabilitating soils for non-agricultural needs but cannot be used for upgrading strongly contaminated soils. The reason is that the sludge addition increases the soil As, Ca, Cr, Mn, and Zn quantities as well as the easily mobilized fractions of the sludge/soil mixtures that contain Cd, Cr, Mg, Mn, Pb, and Tl. The addition of the communal sludge into soils increases the value of the environmental risk in the case of Pb, Mn, Mg, Fe, Cr, Zn, and Ca, but within the same risk classification ranges.