Impacts of Endemic Earthworms (Megascolecidae) in Biosolids-Amended Soil.

Biosolids can be a valuable fertilizer for agriculture and in ecological restoration, although there are concerns about contaminants. Earthworm activity, including vermicomposting of biosolids, may influence the efficacy of their use. We investigated how two New Zealand endemic anecic species of (cf. ) responded to biosolids amendment and affected the mobility of nutrients and trace elements as well as greenhouse gas emissions in biosolids-amended soil. Earthworms were incubated with mixtures of biosolids-amended soil (0, 6.25, 12.5, 25, and 50% biosolids by volume) for 21 d. All species survived in the soil-biosolids mixtures but not in 100% biosolids. The native earthworms, and sp.2, increased KCl-extractable NH and NO by up to 29%, substantially more than . All species significantly increased microbial biomass carbon and Ca(NO)-extractable Cu but significantly decreased dehydrogenase enzymes activity in biosolids-amended soil. Concentrations of Ca(NO)-extractable Mg, S, Fe, Mn, Cd, Co, and Zn varied between earthworm species and with biosolids addition rates. New Zealand native earthworms exacerbated NO emissions from soil, whereas did not. is clearly a preferred species for vermicomposting biosolids and is more tolerant of high concentrations of biosolids. However, New Zealand native earthworms may be more suitable for improving the fertility of soil amended with biosolids.

Potential Environmental Benefits from Blending Biosolids with Other Organic Amendments before Application to Land.

Biosolids disposal to landfill or through incineration is wasteful of a resource that is rich in organic matter and plant nutrients. Land application can improve soil fertility and enhance crop production but may result in excessive nitrate N (NO-N) leaching and residual contamination from pathogens, heavy metals, and xenobiotics. This paper evaluates evidence that these concerns can be reduced significantly by blending biosolids with organic materials to reduce the environmental impact of biosolids application to soils. It appears feasible to combine organic waste streams for use as a resource to build or amend degraded soils. Sawdust and partially pyrolyzed biochars provide an opportunity to reduce the environmental impact of biosolids application, with studies showing reductions of NO-N leaching of 40 to 80%. However, other organic amendments including lignite coal waste may result in excessive NO-N leaching. Field trials combining biosolids and biochars for rehabilitation of degraded forest and ecological restoration are recommended.

Potential Use of Biosolids to Reforest Degraded Areas with New Zealand Native Vegetation.

Biosolids could potentially be used for reforestation of degraded soils in New Zealand with native vegetation. Many native plant species of New Zealand thrive in low-fertility soils, and there is scant knowledge about their nutrient requirements. Therefore, it is unclear whether they will respond positively to the addition of biosolids. We used a pot trial to determine the responses of 11 native plant species to biosolids addition (10% w/w, ∼90 Mg hm) on two distinct degraded soils, Lismore stony silt loam and a Kaikoura sand. We also intended to prove that the soil microbial activity improves with the addition of biosolids, depending on the plant species. All species grew better in Lismore stony silt loam than the Kaikoura sand. All species in the Lismore stony silt loam responded positively to biosolids. The response to biosolids addition in the Kaikoura sand was variable, with four species showing no improvement in growth when biosolids were added. The nutrient status (N, P, S, Cu, and Zn) of all species improved when the two soils were amended with biosolids. However, some plant species, especially Sol. ex Gaertn. and Raoul, showed concerning concentrations of Cd (up to 2.4 mg kg). Dehydrogenase activity of soils (indicator of soil microbial activity) increased in biosolids-amended soils, with a strong species effect. Future work should involve field trials to determine the effect of biosolids addition on the establishment of native plant communities.

Effect of Pine Waste and Pine Biochar on Nitrogen Mobility in Biosolids.

Humanity produces ∼27 kg of dry matter in biosolids per person per year. Land application of biosolids can improve crop production and remediate soils but may result in excessive nitrate N (NO-N) leaching. Carbonaceous materials can reduce the environmental impact of biosolids application. We aimed to ascertain and compare the potentials for Monterey pine ( D. Don)-sawdust-derived biochars and raw sawdust to reduce NO-N leaching from biosolids. We used batch sorption experiments 1:10 ratio of material to solution (100 mg kg of NH or NO) and column leaching experiments with columns containing biosolids (2.7% total N, 130 mg kg NH and 1350 mg kg NO) mixed with soil, biochar, or sawdust. One type of low-temperature (350°C) biochar sorbed 335 mg kg NH, while the other biochars and sawdust sorbed <200 mg kg NH. None of the materials sorbed NO. Biochar added at rates of 20 to 50% reduced NH-N (<1% of total N) leaching from columns by 40 to 80%. Nitrate leaching (<7% of total N) varied little with biochar form or rate but was reduced by sawdust. Incorporating dried sawdust with biosolids showed promise for mitigating NO-N leaching. This effect likely is due to sorption into the pores of the biochar combined with denitrification and immobilization of N rather than chemical sorption onto surfaces.

Biowaste Mixtures Affecting the Growth and Elemental Composition of Italian Ryegrass ().

Biosolids (sewage sludge) can be beneficially applied to degraded lands to improve soil quality. Plants grown on biosolids-amended soils have distinct concentrations of macronutrients and trace elements, which can be beneficial or present a risk to humans and ecosystems. Potentially, biosolids could be blended with other biowastes, such as sawdust, to reduce the risks posed by rebuilding soils using biosolids alone. We sought to determine the effect of mixing biosolids and sawdust on the macronutrient and trace element concentration of ryegrass over a 5-mo period. was grown in a low fertility soil, typical for marginal farm areas, that was amended with biosolids (1250 kg N ha), biosolids + sawdust (0.5:1) and urea (200 kg N ha), as well as a control. Biosolids increased the growth of from 2.93 to 4.14 t ha. This increase was offset by blending the biosolids with sawdust (3.00 t ha). Urea application increased growth to 4.93 t ha. The biowaste treatments increased N, P, Cu, Mn, and Zn relative to the control, which may be beneficial for grazing animals. Although biowaste application caused elevated Cd concentrations (0.15-0.24 mg kg) five- to eightfold higher than control and urea treatments, these were below levels that are likely to result in unacceptable concentrations in animal tissues. Mixing biosolids with sawdust reduced Cd uptake while still resulting in increased micronutrient concentrations (P, S, Mn, Zn, Cu) in plants. There were significant changes in the elemental uptake during the experiment, which was attributed to the decomposition of the sawdust.

Co-Selection of Bacterial Metal and Antibiotic Resistance in Soil Laboratory Microcosms.

Accumulation of heavy metals (HMs) in agricultural soil following the application of superphosphate fertilisers seems to induce resistance of soil bacteria to HMs and appears to co-select for resistance to antibiotics (Ab). This study aimed to investigate the selection of co-resistance of soil bacteria to HMs and Ab in uncontaminated soil incubated for 6 weeks at 25 °C in laboratory microcosms spiked with ranges of concentrations of cadmium (Cd), zinc (Zn) and mercury (Hg). Co-selection of HM and Ab resistance was assessed using plate culture on media with a range of HM and Ab concentrations, and pollution-induced community tolerance (PICT) assays. Bacterial diversity was profiled via terminal restriction fragment length polymorphism (TRFLP) assay and 16S rDNA sequencing of genomic DNA isolated from selected microcosms. Based on sequence data, the microbial communities exposed to HMs were found to differ significantly compared to control microcosms with no added HM across a range of taxonomic levels.

Amplification of oral streptococcal DNA from human incisors and bite marks.

Challenges to the evidentiary value of morphometric determinations have led to a requirement for scientifically substantiated approaches to the forensic analysis of bite marks. Human teeth support genotypically distinctive populations of bacteria that could be exploited for forensic purposes. This study explored the feasibility of directly amplifying bacterial DNA from bite marks for comparison with that from teeth. Samples from self-inflicted experimental bite marks (n = 24) and human incisors were amplified by PCR using primers specific for streptococcal 16S ribosomal DNA. Amplicon profiles (resolved by denaturing gradient gel electrophoresis) from bite mark samples aligned significantly more closely with profiles generated from the teeth responsible than with those from other teeth. Streptococcal amplicons were generated from dental samples applied to excised porcine skin for up to 48 h. These findings indicate that streptococcal DNA can be amplified directly from bite marks, and have potential application in bite mark analysis.

Chemical Elements and the Quality of Manuka (Leptospermum scoparium) Honey.

Soil properties in the foraging range of honeybees influence honey composition. We aimed to determine relationships between the antimicrobial properties of New Zealand manuka (Leptospermum scoparium) honey and elemental concentrations in the honey, plants, and soils. We analyzed soils, plants, and fresh manuka honey samples from the Wairarapa region of New Zealand for the chemical elements and the antimicrobial activity of the honey as indicated by methylglyoxal (MGO) and dihydroxyacetone (DHA). There were significant negative correlations between honey MGO and the concentrations of Mn, Cu, Mg, S, Na, Ba, K, Zn, and Al. These elements may provide a low-cost means of assessing manuka honey quality. For individual elements, except for K, there were no correlations between the honeys, plants, and soils. Soil nitrate concentrations were negatively correlated with concentrations of MGO and DHA in the honey, which implies that soil fertility may be a determiner of manuka honey quality.

PCR-based detection of salivary bacteria as a marker of expirated blood.

Distinguishing between bloodstains caused by a spatter pattern or by expirated blood may be crucial to a forensic investigation. Expirated blood is likely to be contaminated with saliva but current techniques have limited sensitivity, especially with small bloodstains. We report that a PCR assay, designed to detect salivary bacteria, can amplify streptococcal DNA from saliva stains applied to fabrics for at least 62 days after seeding. Bacterial DNA was detected when 0.01 microl of saliva was present in the stain and the amplification was not affected by contamination with blood. These findings indicate that PCR amplification of salivary microbial DNA may have application in the identification of expirated bloodstains in forensic case-work.

Response of a Pioneering Species (Leptospermum scoparium J.R.Forst. & G.Forst.) to Heterogeneity in a Low-Fertility Soil.

Root foraging may increase plant nutrient acquisition at the cost of reducing the total volume of soil explored, thereby reducing the chance of the roots encountering additional patches. Patches in soil seldom contain just one nutrient: the patch may also have distinct textural, hydrological, and toxicological characteristics. We sought to determine the characteristics of root foraging by a pioneering species, Leptospermum scoparium, using pot trials and rhizobox experiments with patches of biosolids. The growth of L. scoparium was increased by <50 t/ha equiv. of biosolids but higher doses were inhibitory. Roots foraged patches of biosolids in a low-fertility soil. There was no evidence of chemotaxis, rather, the roots proliferated toward the patch of biosolids, following chemical gradients of nitrate. While the biosolids also contained high concentrations of other nutrients (P, K, and S), only significant chemical gradients of nitrate were found. Once the roots encountered a patch of biosolids, the growth of the plant increased to a level similar to plants growing in soil homogeneously mixed with biosolids or surface-applied biosolids. Our results indicate that roots forage nitrate, which is mobile in soil, and that gradients of nitrate may lead to patches containing other less mobile nutrients, such as phosphate or potassium.

Investigation of triclosan contamination on microbial biomass and other soil health indicators.

Triclosan (TCS) is an antimicrobial compound found in personal care products, and consequently in greywater. After its release to the environment, it continues its antimicrobial action on indigenous microbial communities. Little is known about the environmental impacts of high levels of TCS, which may occur due to accumulation following long-term greywater application to soil. Soil microcosms were established using a silty clay loam and augmented with a range of TCS concentrations ranging from 500 to 7500 mg kg-1. Samples were analysed for substrate-induced respiration, microbial biomass and sulphatase activity. The soil augmented with the lowest concentration of TCS (500 mg kg-1) significantly decreased microbial biomass, with a calculated EC20 of 195 mg kg-1. Substrate-induced respiration indicated that the soil microbial community was impacted for all TCS concentrations; however, the community showed potential to recover over time. Sulphatase activity was less sensitive to TCS and was significantly impacted at high concentrations of TCS (>2500 mg kg-1). It is likely that TCS has selective toxicity for more susceptible microbes when introduced into the soil environment. At high levels, TCS could overwhelm TCS-degrading soil microbes.

Effects of long-term greywater disposal on soil: A case study.

This study investigated the environmental health risks to soil and potential risks to groundwater associated with long term (8-18years) greywater disposal practices. Land application of greywater is likely to have environmental impacts, which may be positive or negative. Greywater can contain plant macronutrients that may benefit plant growth. Conversely, high levels of surfactants, oils, grease, sodium and potentially pathogenic organisms may negatively impact environmental and human health. In this study, land disposal of untreated greywater was practiced at five coastal domestic properties. At each property, soil samples were collected at two depths from areas used for greywater disposal and from control areas that were not exposed to greywater. Soils were analysed for chemical and biological responses to greywater exposure. Generally, greywater irrigated soils had higher pH, Olsen P, base saturation, and increased soil microbial activity (as measured by biomass carbon, basal respiration and dehydrogenase activity). A pH of >9 was recorded for some greywater treated soil samples. Escherichia coli (E. coli) were detected at up to 10(3)MPN/g in the greywater exposed surface soils at some sites. Terminal Restriction Fragment Length Polymorphism (TRFLP) analysis revealed that greywater affected the soil microbial community structure, which may have implications for soil health and fertility. Overall, this study shows that the long-term application of greywater at the investigated sites had a moderate impact on the soil environment. This may have been due to the sandy soils and high rainfall that would flush the soil. Increases in microbial biomass and dehydrogenase indicate that greywater application may be beneficial for plant growth. However, high levels of E. coli in some soils may be a risk to human health and sub-surface irrigation should be the recommended application method.

The effect of lignite on nitrogen mobility in a low-fertility soil amended with biosolids and urea.

Lignite has been proposed as a soil amendment that reduces nitrate (NO3(-)) leaching from soil. Our objective was to determine the effect of lignite on nitrogen (N) fluxes from soil amended with biosolids or urea. The effect of lignite on plant yield and elemental composition was also determined. Batch sorption and column leaching experiments were followed by a lysimeter trial where a low fertility soil was amended with biosolids (400 kg N/ha equivalent) and urea (200 kg N/ha equivalent). Treatments were replicated three times, with and without lignite addition (20 t/ha equivalent). Lignite did not reduce NO3(-) leaching from soils amended with either biosolids or urea. While lignite decreased NO3(-) leaching from an unamended soil, the magnitude of this effect was not significant in an agricultural context. Furthermore, lignite increased cumulative N2O production from soils receiving urea by 90%. Lignite lessened the beneficial growth effects of adding biosolids or urea to soil. Further work could investigate whether coating urea granules with lignite may produce meaningful environmental benefits.

Ecological impacts of long-term application of biosolids to a radiata pine plantation.

Assessment of the ecological impact of applying biosolids is important for determining both the risks and benefits. This study investigated the impact on soil physical, chemical and biological properties, tree nutrition and growth of long-term biosolids applications to a radiata pine (Pinus radiata D. Don) plantation growing on a Sandy Raw Soil in New Zealand. Biosolids were applied to the trial site every 3 years from tree age 6 to 19 years at three application rates: 0 (Control), 300 (Standard) and 600 (High) kg nitrogen (N) ha(-1), equivalent to 0, 3 and 6 Mg ha(-1) of dry biosolids, respectively. Tree nutrition status and growth have been monitored annually. Soil samples were collected 13 years after the first biosolids application to assess the soil properties and functioning. Both the Standard and High biosolids treatments significantly increased soil (0-50 cm depth) total carbon (C), N, and phosphorus (P), Olsen P and cation exchange capacity (CEC), reduced soil pH, but had no significant effects on soil (0-20 cm depth) physical properties including bulk density, total porosity and unsaturated hydraulic conductivity. The High biosolids treatment also increased concentrations of soil total cadmium (Cd), chromium (Cr), copper (Cu) and lead (Pb) at 25-50 cm depth, but these concentrations were still considered very low for a soil. Ecotoxicological assessment showed no significant adverse effects of biosolids application on either the reproduction of springtails (Folsomia candida) or substrate utilisation ability of the soil microbial community, indicating no negative ecological impact of bisolids-derived heavy metals or triclosan. This study demonstrated that repeated application of biosolids to a plantation forest on a poor sandy soil could significantly improve soil fertility, tree nutrition and pine productivity. However, the long-term fate of biosolids-derived N, P and litter-retained heavy metals needs to be further monitored in the receiving environment.

Use of biosensors to screen urine samples for potentially toxic chemicals.

Forensic toxicology laboratories are often required to implicate or exclude poisoning as a factor in a death or unexplained illness. An analytical tool which enables toxicologists to screen a wide variety of common poisons would be extremely useful. In this paper, we describe the use of a bacterial biosensor for detecting the presence of commonly encountered potentially toxic chemicals in urine. The biosensor responds to any chemical that causes metabolic stress to the bacterial cell and the response is in direct proportion to the concentration of the stressor. This allows a measure of the concentration of a toxicant in urine, without knowing exactly what the toxic compound(s) may be. This affords a distinct advantage over conventional analytical techniques, which require an extensive screening program before it is even known that a toxic compound is present. This preliminary investigation has shown that this biosensor can indicate the presence, in urine, of herbicides such as glyphosate, 2,4-dichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxyacetic acid; the biocide pentachlorophenol; or inorganic poisons such as arsenic, mercury, and cyanide. The biosensor was also shown to be sensitive to a concentration range of these toxicants likely to be found in samples submitted for toxicological analysis.

Forensic comparison of soils by bacterial community DNA profiling.

This preliminary investigation has shown that a soil microbial community DNA profile can be obtained from the small sample of soil recovered from the sole of a shoe, and from soil stains on clothing. We have also shown that these profiles are representative of the site of collection and therefore could potentially be used as associative evidence to prove a link between suspects and crime scenes. Soil community profiles were obtained using the T-RFLP fingerprinting method that uses fluorescent primer technology and semi-automated analysis techniques similar to those used in human DNA profiling in forensic laboratories.

Discrimination of soils at regional and local levels using bacterial and fungal T-RFLP profiling.

DNA profiling of microbial communities has been proposed as a tool for forensic comparison of soils, but its potential to discriminate between soils from similar land use and/or geographic location has been largely unexplored. We tested the ability of terminal restriction fragment length polymorphism (T-RFLP) to discriminate between soils from 10 sites within the Greater Wellington region, New Zealand, based on their bacterial and fungal DNA profiles. Significant differences in bacterial and fungal communities between soils collected from all but one pair of sites were demonstrated. In some instances, specific terminal restriction fragments were associated with particular sites. Patch discrimination was evident within several sites, which could prove useful for site-specific matching (e.g., matching shoe/car tire print to an object). These results support the need for further understanding of the spatial distribution of soil microbial communities before DNA profiling of soil microbial communities can be applied to the forensic context.

A preliminary investigation into the use of biosensors to screen stomach contents for selected poisons and drugs.

The bioluminescence response of two genetically modified (lux-marked) bacteria to potentially toxic compounds (PTCs) in stomach contents was monitored using an in vitro assay. Cells of Escherichia coli HB101 and Salmonella typhimurium both carrying the lux light producing gene on a plasmid (pUDC607) were added to stomach contents containing various concentrations of organic and inorganic compounds. There was some variability in the response of the two biosensors, but both were sensitive to the herbicides glyphosate, 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T); pentachlorophenol (PCP), and inorganic poisons arsenic and mercury at a concentration range likely to be found in stomach contents samples submitted for toxicological analysis. This study demonstrates that biosensor bioassays could be a useful preliminary screening tool in forensic toxicology and that such a toxicological screening should include more than one test organism to maximise the number of PTC's detected. The probability of false positive results from samples containing compounds that may interfere with the assay such as over-the-counter (OTC) drugs and caffeine in tea and coffee was also investigated. Of the substances tested only coffee has the potential to cause false positive results.