Assessing the potential chronic, sublethal and lethal ecotoxicity of land-applying biosolids on Folsomia candida and Lumbricus terrestris.

The ecotoxicity of biosolids has been studied extensively using single-compound toxicity testing and 'spiking' studies; however, little knowledge exists on the ecotoxicity of biosolids as they are land-applied in the Canadian context. The purpose of this study is to elucidate the chronic, sub-lethal (i.e., behavioural), and lethal impacts of land- applying biosolids on the environmentally relevant Folsomia candida (springtails) and Lumbricus terrestris (earthworms) and concomitantly ascertain whether the use of biosolids for nutrient amendment is a sustainable practice. This study is part of a larger multi-compartment programme which includes terrestrial plants and aquatic arthropods. After a review of existing government protocols and research, the current study suggests new environmentally relevant bioassays as to elucidate the true nature of the potential ecotoxicity of land-applying biosolids, within a laboratory context. Specifically, protocols were developed (e.g., shoebox bioassays for L. terrestris sub-lethal testing) or modified (e.g., using Evans' boxes (Evans 1947) for chronic and sub-lethal testing on L. terrestris). Subsequently, two biosolids were tested on springtails and earthworms using avoidance and reproductive bioassay endpoints, at application rates that represent standard (8 tonnes ha-1) and worst-case scenarios (22 tonnes ha-1). Results indicated no effect of biosolids at the environmentally relevant concentration; the worst-case scenario exhibited a positive significantly significant relationship (indicating preference for treatment conditions). We suggest that further assessment of the potential ecotoxicological impact of biosolids employ (i) environmentally relevant organisms, (ii) appropriate bioassays including the use of whole-organism endpoints, and (iii) multi-kingdom testing (e.g., Kingdom Plantae, Animalia) to comprehensively elucidate answers. Lastly, in situ (field assays) are strongly encouraged for future studies.

Nitrogen and Phosphorus Loss Potential from Biosolids-Amended Soils and Biotic Response in the Receiving Water.

Application of municipal biosolids to agricultural soil can improve soil quality and improve crop yields. However, runoff or tile leachate from biosolids-applied fields may contribute to localized eutrophication of surface water. A laboratory experiment was conducted to determine loss potential of nutrients from soils amended with two different biosolids (anaerobically digested and chemically stabilized) relative to loss from a reference soil and to determine response in freshwater microcosms to nutrients lost from soils. Total phosphorus (TP) and total nitrogen (TN) were measured in runoff, and equivalent amounts were added to reference microcosms to determine if aquatic systems would respond similarly to TN and TP loading in bioavailable forms (PO, NH, NO) simulating loading related to inorganic fertilizer application. Nutrient concentrations (TP, TN, PO, NH, NO, and organic P and N) were similar in the runoff from the two biosolids-amended soils and higher than those in the runoff from the reference soil. Runoff from biosolids-amended soils stimulated algal growth and production (chlorophyll a and dissolved oxygen) relative to runoff from reference soil, but the response was weaker than in microcosms receiving equivalent amounts of inorganic N and P. Nutrient runoff from land-applied biosolids does have potential to increase algal production in receiving waters; however, this experiment suggests receiving waters may absorb a single large nutrient loading event associated with runoff from biosolids-amended soil without substantial impact. Moreover, the response to N and P in biosolids versus inorganic nutrient additions suggests biosolids may contribute relatively less to eutrophication than inorganic fertilizers, assuming equivalent TN and TP loading to aquatic systems.

Utilization of multiple organisms in a proposed early-warning biomonitoring system for real-time detection of contaminants: preliminary results and modeling.

During past decades, biomonitors were deployed in lakes and rivers to rapidly detect hazardous chemicals by measuring the endpoints of a single aquatic species at defined short intervals. Most biomonitors, however, are only capable of indicating a departure from baseline water conditions without identifying the cause. In order to provide a more comprehensive assessment, a biomonitoring system which features a library of stereotyped responses of multiple aquatic species in various water conditions is proposed. A preliminary library was constructed by characterizing the behavioural and physiological responses of Daphnia magna, Hyalella azteca, Lumbriculus variegatus, and Pseudokirchneriella subcapitata to various concentrations of atrazine and tributyltin. By employing multivariate statistical tools such as principal component analysis (PCA) and discriminant analysis, this library (which contained responses after 6h of exposure to contaminants) was used as a template to classify and to model other sets of earlier measurements at 2 and 4h, resulting in an accuracy of 73 and 97%, respectively. These findings demonstrated the potential capability of the proposed early-warning biomonitoring system to provide real-time water quality assessment and early-warning contaminant detection.

Evaluation of low-copy genetic targets for waterborne bacterial pathogen detection via qPCR.

Recent developments in water quality research have highlighted difficulties in accurately predicting the incidence of pathogens within freshwater based on the viability, culturability and metabolic activity of indicator organisms. QPCR-driven assays are candidates to replace standard culture-based methods, however, protocols suitable for routine use have yet to be sufficiently validated. The objective of this study was to evaluate five oligonucleotide primers sets (ETIR, SINV, exoT, VS1 and ipaH2) for their potential applicability in qPCR assays to detect contamination from five waterborne bacterial pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Campylobacter jejuni, Pseudomonas aeruginosa, and Shigella flexneri). An enrichment-free qPCR protocol was also tested using S. Typhimurium-seeded source water, combining membrane filtration and mechanical, chemical and enzymatic lysis techniques to recover the bacterial cells. All five primer sets were found to have high specificity and sensitivity for the tested organisms. Four of the primers were able to detect pathogen loads as low as 10 cells/mL while 200 cells/mL of C. jejuni were detectable in pure culture. Although sensitivity decreased in an artificially contaminated environmental matrix, it was still possible to detect as few as 10 S. Typhimurium cells without enrichment. The primers and protocols evaluated in this study have demonstrated potential for further validation for possible application alongside traditional indicator techniques.

Assessing the impact of land-applied biosolids from a thermomechanical (TMP) pulp mill to a suite of terrestrial and aquatic bioassay organisms under laboratory conditions.

The potential impact on a variety of bioassay organisms when pulp-mill biosolids from a thermomechanical pulp mill (western Canada) were applied to a reference soil has been investigated in a laboratory setup. The current research assessed acute, chronic, and reproductive impacts using a battery of terrestrial and aquatic organisms. Terrestrial organisms were exposed to soil amended with different concentrations of biosolids, while aquatic organisms were used to assess the impact of biosolids' runoff into receiving waters. The former bioassays showed that an application rate of 20 tonneshectare(-1) (tha(-1)) "bone-dry" biosolids applied to reference soil produced no observable adverse impact on the terrestrial organisms. In the latter assays, undiluted (100%) and 50% diluted biosolids' runoff into receiving water had a detrimental impact on the aquatic organisms. However, concentrations not exceeding 25% (environmentally relevant concentrations) had neither an acute nor chronic impact compared to reference populations. The organisms' abilities to reproduce were also unaltered. While this study only examined the biosolids from one mill, there is the potential that land-application of characteristically well-defined pulp mill biosolids may constitute an acceptable way of disposing of pulp and paper mill biosolid residues. However, the biosolids coming from different mills, with differing processes, must be dealt with on a case-by-case situation. Each series of biosolids must be rigorously tested for toxicological impact in the laboratory under tightly controlled conditions. Subsequently, field experimentation must be conducted before definitive conclusions can be made.

In vivo and in vitro assessment of the androgenic potential of a pulp and paper mill effluent.

The androgenic potential of a New Zealand pulp and paper mill effluent was measured by applying a combination of in vitro and in vivo bioassays with mosquitofish (Gambusia affinis) and goldfish (Carassius auratus). The in vivo method assessed the rate of gonopodial development (masculinization) and alterations from normal reproductive behavior in adult female mosquitofish exposed for 21 d to untreated or secondary-treated pulp mill effluent. A second in vivo mosquitofish exposure tested the effect of glass-fiber (type C) filtration of secondary-treated effluent on rates of expression of the same endpoints. Extractable organics analyses of effluents and extracts thereof were conducted. Mosquitofish demonstrated significant masculinization on exposure to either treated or untreated effluent; the frequency of gonopodial development was reduced with effluent secondary-treatment. Male mating behavior was observed in the masculinized adult females. Glass-fiber (type F) filtration of the treated effluent eliminated the masculinizing effect, suggesting that the bioactive compounds were associated with the suspended solids. The in vitro method measured the binding of compounds within a treated thermomechanical/bleached kraft effluent extract to androgen receptors contained in goldfish testis cytosol. Exposure to extracts of either the particulate (glass-fiber filtered) or the dissolved organic fraction of the effluent produced significant binding (as indicated by the displacement of radiolabeled testosterone) to the androgen receptor in goldfish gonadal tissue. Thus, the dissolved organics extract of the treated effluent contained compounds androgenic to goldfish in vitro but not to mosquitofish in vivo. The combined in vitro and in vivo data suggest that the effluent in question could exert effects on the reproductive physiology of fishes through an androgenic mechanism. The androgenic compounds androstenedione and testosterone were not detected in the extracts used for the in vitro component of this study.