Three novel multiplex PCR assays for rapid detection of virulence, antimicrobial resistance, and toxin genes in Acinetobacter calcoaceticus-baumannii complex species.

The Acinetobacter calcoaceticus-baumannii (ACB) complex is an often-overlooked group of nosocomial pathogens with a significant environmental presence. Rapid molecular screening methods for virulence, antimicrobial resistance, and toxin (VAT) genes are required to investigate the potential pathogenicity of environmental isolates. This study aimed to develop and apply novel ACB complex-specific multiplex PCR (mPCR) primers and protocols for the rapid detection of eight VAT genes. We optimized three single-tube mPCR assays using reference DNA from ACB complex and other Acinetobacter species. These assays were then applied to detect VAT genes in cultured ACB complex isolates recovered from clinical and environmental sources. Widespread detection of VAT genes in environmental isolates confirmed the validity, functionality, and applicability of these novel assays. Overall, the three newly developed ACB complex species-specific mPCR assays are rapid and simple tools that can be adopted in diagnostic and clinical lab settings. The detection of VAT genes in environmental isolates suggests that environmental niches could serve as a reservoir for potentially pathogenic ACB complex and warrants further investigation. The newly developed mPCR assays are specific, sensitive, and efficient, making them well-suited for high-throughput screening in epidemiological studies and evaluating the potential pathogenicity of ACB complex recovered from various sources.

Modeling West Nile Virus transmission in birds and humans: Advantages of using a cellular automata approach.

In Canada, the periodic circulation of West Nile Virus (WNV) is difficult to predict and, beyond climatic factors, appears to be related to the migratory movements of infected birds from the southern United States. This hypothesis has not yet been explored in a spatially distributed model. The main objective of this work was to develop a spatially explicit dynamic model for the transmission of WNV in Canada, that allows us to explore non-climate related hypotheses associated with WNV transmission. A Cellular Automata (CA) approach for multiple hosts (birds and humans) is used for a test region in eastern Ontario, Canada. The tool is designed to explore the role of host and vector spatial heterogeneity, host migration, and vector feeding preferences. We developed a spatialized compartmental SEIRDS-SEI model for WNV transmission with a study region divided into 4 km2 rectangular cells. We used 2010-2021 bird data from the eBird project and 2010-2019 mosquito data collected by Ontario Public Health to mimic bird and mosquito seasonal variation. We considered heterogeneous bird densities (high and low suitability areas) and homogeneous mosquito and human densities. In high suitability areas for birds, we identified 5 entry points for WNV-infected birds. We compared our simulations with pools of WNV-infected field collected mosquitoes. Simulations and sensitivity analyses were performed using MATLAB software. The results showed good correspondence between simulated and observed epidemics, supporting the validity of our model assumptions and calibration. Sensitivity analysis showed that a 5% increase or decrease in each parameter of our model except for the biting rate of bird by mosquito (c(B,M)) and mosquito natural mortality rate (dM), had a very limited effect on the total number of cases (newly infected birds and humans), prevalence peak, or date of occurrence. We demonstrate the utility of the CA approach for studying WNV transmission in a heterogeneous landscape with multiple hosts.

Local and regional-scale effects of hedgerows on grassland- and forest-associated bird populations within agroecosystems.

Linear woody features (LWFs), like hedgerows along field edges, provide wildlife habitat and support biodiversity in agroecosystems. Assessments of LWFs usually focus on community-level indices, such as species richness. However, effective conservation actions need to balance the contrasting habitat preferences of different wildlife species, necessitating a focus on population-level effects in working landscapes. We assessed associations between LWFs and abundance for 45 bird species within an intensive agroecosystem in eastern Ontario, Canada. We used distance- and removal-sampling methods across 4 years (2016-2019) to estimate local bird abundance in habitats representing a range of LWF densities. We also predicted abundance across a subset of the study region with and without LWFs to understand their contribution to regional population density. Associations between local bird abundance and LWFs were variable among species, but overall community effects were clearly positive, particularly for forest and shrubland species. At the site level, 20/45 species (44%) had higher densities associated with greater LWF presence on average, compared to 5/45 (11%) with negative associations. At the regional scale, LWFs had predicted benefits on total abundance for 31 species (69%), contributing to an estimated 20% increase on average. Positive effects were most pronounced in areas with greater agricultural land use (primarily field crops), suggesting LWFs may provide crucial habitat in heavily modified landscapes but have little to no additional benefit for the avian community in areas with greater existing heterogeneity and habitat retention. Species that responded negatively tended to be at risk with strong habitat preferences for intact forests or large, open grasslands and, thus, greater sensitivity to potential edge effects. With rapidly declining songbird populations and a global need for food security, conservation strategies that amplify biodiversity and enhance agricultural productivity through ecosystem services such as pest control, pollination, and water regulation are vital. We demonstrate the benefits of habitat heterogeneity in agroecosystems on songbird densities and highlight the need to integrate local and landscape-level assessments in conservation planning. An effective, balanced strategy includes concentrating LWFs in areas of extensive arable crops, with habitat retention patches where possible, while maintaining heterogeneity through mixtures of natural habitats and pastoral farming in less intensive regions.

Les caractéristiques d'emboisement linéaires (CEL), tels que les haies en bordure de champs, fournissent un habitat à la faune et à la flore et favorisent la biodiversité dans les agroécosystèmes. Les évaluations des caractéristiques d'emboisement linéaires se concentrent généralement sur des indices au niveau de la communauté, tels que la richesse des espèces. Cependant, pour être efficaces, les actions de conservation doivent équilibrer les préférences variables des différentes espèces sauvages en matière d'habitat, ce qui nécessite de se concentrer sur les effets au niveau de la population dans les paysages exploités. Nous avons évalué les associations entre les CEL et l'abondance de 45 espèces d'oiseaux dans un agroécosystème intensif de l'est de l'Ontario, au Canada. Nous avons utilisé des méthodes d'échantillonnage par distance et par enlèvement sur quatre ans (2016­2019) pour estimer l'abondance locale des oiseaux dans des habitats représentant une gamme de densités de CEL. Nous avons également prédit l'abondance dans un sous­ensemble de la région étudiée avec et sans CEL pour comprendre leur contribution à la densité de la population régionale. Les associations entre l'abondance des oiseaux locaux et les CEL étaient variables d'une espèce à l'autre, mais les effets globaux sur les communautés étaient clairement positifs, en particulier pour les espèces des forêts et des zones arbustives. Au niveau du site, 20/45 espèces (44%) avaient des densités plus élevées associées à une plus grande présence de CEL en moyenne, contre 5/45 (11%) avec des associations négatives. À l'échelle régionale, les CEL ont eu des effets bénéfiques sur l'abondance totale de 31 espèces (69%), contribuant à une augmentation estimée à 20% en moyenne. Les effets positifs étaient plus prononcés dans les zones où l'utilisation des terres agricoles était plus importante (principalement les grandes cultures), ce qui suggère que les CEL peuvent fournir un habitat crucial dans les paysages fortement modifiés, mais qu'elles ont peu ou pas d'avantages supplémentaires pour la communauté aviaire dans les zones où l'hétérogénéité existante et la conservation de l'habitat sont plus importantes. Les espèces qui ont répondu négativement avaient tendance à être en danger, avec de fortes préférences d'habitat pour les forêts intactes ou les grandes prairies ouvertes, et donc une plus grande sensibilité aux effets de lisière potentiels. Avec le déclin rapide des populations d'oiseaux chanteurs et le besoin mondial de sécurité alimentaire, les stratégies de conservation qui amplifient la biodiversité et améliorent la productivité agricole grâce aux services écosystémiques tels que la lutte contre les ravageurs, la pollinisation et la régulation de l'eau sont vitales. Nous démontrons les avantages de l'hétérogénéité de l'habitat dans les agroécosystèmes sur les densités de passereaux et soulignons la nécessité d'intégrer les évaluations locales et au niveau du paysage dans la planification de la conservation. Une stratégie efficace et équilibrée consiste à concentrer les CEL dans les zones de cultures arables extensives, avec des parcelles de conservation de l'habitat là où c'est possible, tout en maintenant l'hétérogénéité grâce à des mélanges d'habitats naturels et à l'agriculture pastorale dans les régions moins intensives.

Pathogenicity assessment of Arcobacter butzleri isolated from Canadian agricultural surface water.

BACKGROUND: Water is considered a source for the transmission of Arcobacter species to both humans and animals. This study was conducted to assess the prevalence, distribution, and pathogenicity of A. butzleri strains, which can potentially pose health risks to humans and animals. Cultures were isolated from surface waters of a mixed-use but predominately agricultural watershed in eastern Ontario, Canada. The detection of antimicrobial resistance (AMR) and virulence-associated genes (VAGs), as well as enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) assays were performed on 913 A. butzleri strains isolated from 11 agricultural sampling sites. RESULTS: All strains were resistant to one or more antimicrobial agents, with a high rate of resistance to clindamycin (99%) and chloramphenicol (77%), followed by azithromycin (48%) and nalidixic acid (49%). However, isolates showed a significantly (p < 0.05) high rate of susceptibility to tetracycline (1%), gentamycin (2%), ciprofloxacin (4%), and erythromycin (5%). Of the eight VAGs tested, ciaB, mviN, tlyA, and pldA were detected at high frequency (> 85%) compared to irgA (25%), hecB (19%), hecA (15%), and cj1349 (12%) genes. Co-occurrence analysis showed A. butzleri strains resistant to clindamycin, chloramphenicol, nalidixic acid, and azithromycin were positive for ciaB, tlyA, mviN and pldA VAGs. ERIC-PCR fingerprint analysis revealed high genetic similarity among strains isolated from three sites, and the genotypes were significantly associated with AMR and VAGs results, which highlight their potential environmental ubiquity and potential as pathogenic. CONCLUSIONS: The study results show that agricultural activities likely contribute to the contamination of A. butzleri in surface water. The findings underscore the importance of farm management practices in controlling the potential spread of A. butzleri and its associated health risks to humans and animals through contaminated water.

The role of species ecology in predicting Toxoplasma gondii prevalence in wild and domesticated mammals globally.

Macroecological approaches can provide valuable insight into the epidemiology of globally distributed, multi-host pathogens. Toxoplasma gondii is a protozoan that infects any warm-blooded animal, including humans, in almost every habitat worldwide. Toxoplasma gondii infects its hosts through oocysts in the environment, carnivory of tissue cysts within intermediate host prey and vertical transmission. These routes of infection enable specific predictions regarding the ecological and life history traits that should predispose specific taxa to higher exposure and, thus infection rates of T. gondii. Using T. gondii prevalence data compiled from 485 studies representing 533 free-ranging wild mammalian species, we examined how ecological (habitat type, trophic level) and life history (longevity, vagility, gestation duration and torpor) traits influence T. gondii infection globally. We also compared T. gondii prevalence between wild and domesticated species from the same taxonomic families using data compiled from 540 studies of domestic cattle, sheep, and pigs. Across free-ranging wildlife, we found the average T. gondii prevalence was 22%, which is comparable to the global human estimate. Among ecological guilds, terrestrial species had lower T. gondii prevalence than aquatic species, with freshwater aquatic taxa having an increased prevalence compared to marine aquatic species. Dietary niches were also influential, with carnivores having an increased risk compared to other trophic feeding groups that have reduced tissue cyst exposure in their diet. With respect to influential life history traits, we found that more vagile wildlife species had higher T. gondii infection rates, perhaps because of the higher cumulative risk of infection during movement through areas with varying T. gondii environmental loads. Domestic farmed species had a higher T. gondii prevalence compared to free-ranging confamilial wildlife species. Through a macroecological approach, we determined the relative significance of transmission routes of a generalist pathogen, demonstrating an increased infection risk for aquatic and carnivorous species and highlighting the importance of preventing pathogen pollution into aquatic environments. Toxoplasma gondii is increasingly understood to be primarily an anthropogenically-associated pathogen whose dissemination is enhanced by ecosystem degradation and human subsidisation of free-roaming domestic cats. Adopting an ecosystem restoration approach to reduce one of the world's most common parasites would synergistically contribute to other initiatives in conservation, feline and wildlife welfare, climate change, food security and public health.

A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples.

Mosquito-borne diseases pose ongoing global health concerns, demanding more cost-efficient methods to detect pathogens to support enhanced surveillance efforts. This study introduces an adapted TRIzol-based high-throughput RNA extraction protocol, tailored for the detection of California serogroup viruses in pooled mosquito samples in a rapid and cost-effective manner. This approach provided consistent RNA yields and sensitive viral detection relative to two commercial extraction kits (QIAGEN RNeasy Mini Kit and MACHEREY-NAGEL NucleoSpin RNA Plus Kit). The incorporation of a user-friendly and non-spiking-based RT-qPCR internal control designed for the 18S rRNA gene in mosquitoes minimizes potential false positives/negatives, improving the fidelity of viral detection outcomes. Effective RNA yields, purity, and successful target amplification across 25 mosquito species and varied pool sizes (1-50 mosquitoes per tube) affirm the reliability of our approach. The extraction method is cost-effective, with an incurred cost of $0.58 CAD per sample, in contrast to the $5.25 CAD cost per sample of the two kits, rendering it promising for mosquito-borne disease surveillance initiatives.

Relationships between water quality and mosquito presence and abundance: a systematic review and meta-analysis.

Mosquito-borne diseases (MBDs) are emerging in response to climate and land use changes. As mosquito (Diptera: Culicidae) habitat selection is often contingent on water availability for egg and larval development, studies have recognized water quality also influences larval habitats. However, underlying species-, genera-, and mosquito level preferences for water quality conditions are varied. This systematic review and meta-analysis aimed to identify, characterize, appraise, and synthesize available global data on the relationships between water quality and mosquito presence and abundance (MPA); with the goal to further our understanding of the geographic expansion of MBD risks. A systematic review was conducted to identify studies investigating the relationships between water quality properties and MPA. Where appropriate, random-effects meta-analyses were conducted to provide pooled estimates for the association between the most reported water quality properties and MPA. The most reported water quality parameters were pH (87%), nitrogen concentrations (56%), turbidity (56%), electrical conductivity (54%), dissolved oxygen (43%), phosphorus concentrations (30%), and alkalinity (10%). Overall, pH (P = 0.05), turbidity (P < 0.0001), electrical conductivity (P = 0.005), dissolved oxygen (P < 0.0001), nitrogen (P < 0.0001), and phosphorus (P < 0.0001) showed significantly positive pooled correlations with MPA, while alkalinity showed a nonsignificant null pooled correlation (P = 0.85). We observed high heterogeneity in most meta-analyses, and climate zonation was shown to influence the pooled estimates. Linkages between MPA and water quality properties will enhance our capacity to predict MBD risks under changing environmental and land use changes.

Future land-use change predictions using Dyna-Clue to support mosquito-borne disease risk assessment.

Mosquitoes are known vectors for viral diseases in Canada, and their distribution is driven by climate and land use. Despite that, future land-use changes have not yet been used as a driver in mosquito distribution models in North America. In this paper, we developed land-use change projections designed to address mosquito-borne disease (MBD) prediction in a 38 761 km2 area of Eastern Ontario. The landscape in the study area is marked by urbanization and intensive agriculture and hosts a diverse mosquito community. The Dyna-CLUE model was used to project land-use for three time horizons (2030, 2050, and 2070) based on historical trends (from 2014 to 2020) for water, forest, agriculture, and urban land uses. Five scenarios were generated to reflect urbanization, agricultural expansion, and natural areas. An ensemble of thirty simulations per scenario was run to account for land-use conversion uncertainty. The simulation closest to the average map generated was selected to represent the scenario. A concordance matrix generated using map pair analysis showed a good agreement between the simulated 2020 maps and 2020 observed map. By 2050, the most significant changes are predicted to occur mainly in the southeastern region's rural and forested areas. By 2070, high deforestation is expected in the central west. These results will be integrated into risk models predicting mosquito distribution to study the possibility of humans' increased exposure risk to MBDs.

Environmental Concentrations of the Type 2 Diabetes Medication Metformin and Its Transformation Product Guanylurea in Surface Water and Sediment in Ontario and Quebec, Canada.

Metformin, used to treat Type 2 diabetes, is the active ingredient of one of the most prescribed drugs in the world, with over 120 million yearly prescriptions globally. In wastewater-treatment plants (WWTPs), metformin can undergo microbial transformation to form the product guanylurea, which could have toxicological relevance in the environment. Surface water samples from 2018 to 2020 and sediment samples from 2020 were collected from six mixed-use watersheds in Quebec and Ontario, Canada, and analyzed to determine the metformin and guanylurea concentrations at each site. Metformin and guanylurea were present above their limits of quantification in 51.0% and 50.7% of all water samples and in 64% and 21% of all sediment samples, respectively. In surface water, guanylurea was often present at higher concentrations than metformin, while the inverse was true in sediment, with metformin frequently detected at higher concentrations than guanylurea. In addition, at all sites influenced solely by agriculture, concentrations of metformin and guanylurea were <1 µg/L in surface water, suggesting that agriculture is not a significant source of these compounds in the investigated watersheds. These data suggest that WWTPs and potentially septic system leaks are the most likely sources of the compounds in the environment. Guanylurea was detected at many of these sites above environmental concentrations of concern, where critical processes in fish may be affected. Due to the scarcity of available ecotoxicological data and the prominence of guanylurea across all sample sites, there is a need to perform more toxicological investigations of this transformation product and revisit regulations. The present study will help provide toxicologists with environmentally relevant concentration ranges in Canada. Environ Toxicol Chem 2023;42:1709-1720. © 2023 His Majesty the King in Right of Canada and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

Impacts of repeated coastal flooding on soil and groundwater following managed dike realignment.

Coastal defense structures (e.g., dikes, seawalls) protect vulnerable communities along marine coastlines and estuaries from the physical and chemical influences of adjacent water bodies. These structures are susceptible to overtopping or breaching by tides and waves, with risks amplified by climate change-induced sea-level rise. Repeated inundation by saline water can contaminate freshwater resources and salinize soil, impacting land-use activities, including agricultural productivity. Managed ecosystem-based dike realignment and salt marsh restoration can provide alternatives to traditional coastal adaptation approaches. We assess the changes to soil salinity at a managed dike realignment project prior to the transformation from a diked terrestrial environment to an estuarine environment. Baseline data are compared to conditions following 8-10 months of intermittent flooding at spring tides. Results show that an increase in salinity occurred over the entire site in the shallow subsurface, with the most significant contamination occurring in low-lying areas. Bulk soil electrical conductivity (salinity proxy) measured from geophysical surveys increased from the previous freshwater condition of ∼300 µS/cm to over 6000 µS/cm following <20 flood events, while successive flooding resulted in increased soil moisture as infiltrated floodwater propagated to greater depths. Sediment deposition occurred at high rates, with up to 4 cm of sediment deposited per flood, converting much of the previously cultivated land into tidal mudflats. Deeper sediments and groundwater (i.e., >1.8 m depth) were not impacted over the time scale of this research. This study demonstrates that intermittent shallow flooding can rapidly increase moisture content and soil salinity in surficial sediments and, in turn, adversely impact conditions suitable for agricultural crop production. The realignment zone serves as an engineered analog of coastal flooding, presenting an opportunity to investigate how low-lying coastal environments may experience regular flooding in the future due to sea-level rise and intensifying coastal storms.

How many microplastic particles are present in Canadian biosolids?

Application of treated sewage sludge (biosolids) from wastewater treatment plants (WWTPs) to farmlands is an important pathway through which microplastic particles (MPs) enter terrestrial ecosystems. Yet, microplastic concentrations in Canadian biosolids have only been estimated in samples from four WWTPs previously. We aimed to fill this knowledge gap by quantifying microplastics in biosolids from 22 WWTPs located in nine provinces and two commercial fertilizer producers in Canada. All samples had substantial microplastic concentrations ranging from 228 to 1353 particles per gram dry weight (median = 636 particles), which are orders of magnitude greater than MPs reported from earlier investigations of biosolids from other countries. Fibers (median: 86%) were the most common type of MPs observed, followed by fragments (median: 13%). There were no statistically significant differences in the amount of microplastics observed in the biosolids from different geographical regions, WWTP types, and sludge treatment processes. This suggests that diverse combinations of local sewershed characteristics, site-specific treatment approaches, and daily flow at WWTPs may be influencing concentrations of microplastics in biosolids. Our results indicate that microplastic concentrations in biosolids are substantially higher than they are in other environmental matrices, and this has important implications to managing microplastic pollution in terrestrial ecosystems.

Core and conditionally rare taxa as indicators of agricultural drainage ditch and stream health and function.

BACKGROUND: The freshwater microbiome regulates aquatic ecological functionality, nutrient cycling, pathogenicity, and has the capacity to dissipate and regulate pollutants. Agricultural drainage ditches are ubiquitous in regions where field drainage is necessary for crop productivity, and as such, are first-line receptors of agricultural drainage and runoff. How bacterial communities in these systems respond to environmental and anthropogenic stressors are not well understood. In this study, we carried out a three year study in an agriculturally dominated river basin in eastern Ontario, Canada to explore the spatial and temporal dynamics of the core and conditionally rare taxa (CRT) of the instream bacterial communities using a 16S rRNA gene amplicon sequencing approach. Water samples were collected from nine stream and drainage ditch sites that represented the influence of a range of upstream land uses. RESULTS: The cross-site core and CRT accounted for 5.6% of the total number of amplicon sequence variants (ASVs), yet represented, on average, over 60% of the heterogeneity of the overall bacterial community; hence, well reflected the spatial and temporal microbial dynamics in the water courses. The contribution of core microbiome to the overall community heterogeneity represented the community stability across all sampling sites. CRT was primarily composed of functional taxa involved in nitrogen (N) cycling and was linked to nutrient loading, water levels, and flow, particularly in the smaller agricultural drainage ditches. Both the core and the CRT were sensitive responders to changes in hydrological conditions. CONCLUSIONS: We demonstrate that core and CRT can be considered as holistic tools to explore the temporal and spatial variations of the aquatic microbial community and can be used as sensitive indicators of the health and function of agriculturally dominated water courses. This approach also reduces computational complexity in relation to analyzing the entire microbial community for such purposes.

In silico identification of multiple conserved motifs within the control region of Culicidae mitogenomes.

Mosquitoes are important vectors for human and animal diseases. Genetic markers, like the mitochondrial COI gene, can facilitate the taxonomic classification of disease vectors, vector-borne disease surveillance, and prevention. Within the control region (CR) of the mitochondrial genome, there exists a highly variable and poorly studied non-coding AT-rich area that contains the origin of replication. Although the CR hypervariable region has been used for species differentiation of some animals, few studies have investigated the mosquito CR. In this study, we analyze the mosquito mitogenome CR sequences from 125 species and 17 genera. We discovered four conserved motifs located 80 to 230 bp upstream of the 12S rRNA gene. Two of these motifs were found within all 392 Anopheles (An.) CR sequences while the other two motifs were identified in all 37 Culex (Cx.) CR sequences. However, only 3 of the 304 non-Culicidae Dipteran mitogenome CR sequences contained these motifs. Interestingly, the short motif found in all 37 Culex sequences had poly-A and poly-T stretch of similar length that is predicted to form a stable hairpin. We show that supervised learning using the frequency chaos game representation of the CR can be used to differentiate mosquito genera from their dipteran relatives.

Effects of land use and weather on the presence and abundance of mosquito-borne disease vectors in a urban and agricultural landscape in Eastern Ontario, Canada.

Weather and land use can significantly impact mosquito abundance and presence, and by consequence, mosquito-borne disease (MBD) dynamics. Knowledge of vector ecology and mosquito species response to these drivers will help us better predict risk from MBD. In this study, we evaluated and compared the independent and combined effects of weather and land use on mosquito species occurrence and abundance in Eastern Ontario, Canada. Data on occurrence and abundance (245,591 individuals) of 30 mosquito species were obtained from mosquito capture at 85 field sites in 2017 and 2018. Environmental variables were extracted from weather and land use datasets in a 1-km buffer around trapping sites. The relative importance of weather and land use on mosquito abundance (for common species) or occurrence (for all species) was evaluated using multivariate hierarchical statistical models. Models incorporating both weather and land use performed better than models that include weather only for approximately half of species (59% for occurrence model and 50% for abundance model). Mosquito occurrence was mainly associated with temperature whereas abundance was associated with precipitation and temperature combined. Land use was more often associated with abundance than occurrence. For most species, occurrence and abundance were positively associated with forest cover but for some there was a negative association. Occurrence and abundance of some species (47% for occurrence model and 88% for abundance model) were positively associated with wetlands, but negatively associated with urban (Culiseta melanura and Anopheles walkeri) and agriculture (An. quadrimaculatus, Cs. minnesotae and An. walkeri) environments. This study provides predictive relationships between weather, land use and mosquito occurrence and abundance for a wide range of species including those that are currently uncommon, yet known as arboviruses vectors. Elucidation of these relationships has the potential to contribute to better prediction of MBD risk, and thus more efficiently targeted prevention and control measures.

Monitoring of Environmental Contaminants in Mixed-Use Watersheds Combining Targeted and Nontargeted Analysis with Passive Sampling.

Understanding the environmental fate, transport, and occurrence of pesticides and pharmaceuticals in aquatic environments is of utmost concern to regulators. Traditionally, monitoring of environmental contaminants in surface water has consisted of liquid chromatography-tandem mass spectrometry analyses for a set of targeted compounds in discrete samples. These targeted approaches are limited by the fact that they only provide information on compounds within a target list present at the time and location of sampling. To address these limitations, there has been considerable interest in suspect screening and nontargeted analysis (NTA), which allow for the detection of all ionizable compounds in the sample with the added benefit of data archiving for retrospective mining. Even though NTA can detect a large number of contaminants, discrete samples only provide a snapshot perspective of the chemical disposition of an aquatic environment at the time of sampling, potentially missing episodic events. We evaluated two types of passive chemical samplers for nontargeted analysis in mixed-use watersheds. Nontargeted data were processed using MS-DIAL to screen against our in-house library and public databases of more than 1300 compounds. The data showed that polar organic chemicals integrative samplers (POCIS) were able to capture the largest number of analytes with better reproducibility than organic compound-diffusive gradients in thin film (o-DGT), resulting from the greater amount of binding sorbent. We also showed that NTA combined with passive sampling gives a more representative picture of the contaminants present at a given site and enhances the ability to identify the nature of point and nonpoint pollution sources and ecotoxicological impacts. Environ Toxicol Chem 2022;41:1131-1143. © 2021 Her Majesty the Queen in Right of Canada Environmental Toxicology and Chemistry © 2021 SETAC. Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

Human density is associated with the increased prevalence of a generalist zoonotic parasite in mammalian wildlife.

Macroecological approaches can provide valuable insight into the epidemiology of globally distributed, multi-host pathogens. Toxoplasma gondii is a zoonotic protozoan that infects any warm-blooded animal, including humans, in almost every ecosystem worldwide. There is substantial geographical variation in T. gondii prevalence in wildlife populations and the mechanisms driving this variation are poorly understood. We implemented Bayesian phylogenetic mixed models to determine the association between species' ecology, phylogeny and climatic and anthropogenic factors on T. gondii prevalence. Toxoplasma gondii prevalence data were compiled for free-ranging wild mammal species from 202 published studies, encompassing 45 079 individuals from 54 taxonomic families and 238 species. We found that T. gondii prevalence was positively associated with human population density and warmer temperatures at the sampling location. Terrestrial species had a lower overall prevalence, but there were no consistent patterns between trophic level and prevalence. The relationship between human density and T. gondii prevalence is probably mediated by higher domestic cat abundance and landscape degradation leading to increased environmental oocyst contamination. Landscape restoration and limiting free-roaming in domestic cats could synergistically increase the resiliency of wildlife populations and reduce wildlife and human infection risks from one of the world's most common parasitic infections.

The Effects of Ditch Management in Agroecosystems on Embryonic and Tadpole Survival, Growth, and Development of Northern Leopard Frogs (Lithobates pipiens).

Agricultural drainage ditches help remove excess water from fields and provide habitat for wildlife. Drainage ditch management, which includes various forms of vegetation clearing and sediment dredging, can variably affect the ecological function of these systems. To determine whether ditch conditions following dredging/vegetation clearing management affected the survival, growth, and development of embryos and tadpoles of northern leopard frogs (Lithobates pipiens), we conducted three field studies using in situ cages over 2 years. We measured nutrients, pesticides, and other water quality properties in vegetated/unmanaged (i.e., no clearing or dredging) and newly cleared/dredged (i.e., treeless, then dredged), clay-bottomed drainage ditches in a river basin in Eastern Ontario, Canada. Nutrients, atrazine, and total neonicotinoid concentrations were generally lower at the cleared/dredged sites, whereas glyphosate was at higher concentrations. In contrast, water-quality variables measured in situ, particularly temperature, dissolved oxygen, and turbidity, tended to be higher in the cleared/dredged sites. Total phosphorous and total organic carbon concentrations at all sites were above the recommended limits for amphibian assays. No significant differences were detected in the survival, hatching success, or development of embryos among the ditch management treatments, but premature hatching was observed at one vegetated/unmanaged site where high specific conductivity may have been formative. We found the cleared/dredged sites supported earlier tadpole growth and development, likely as a result of the higher water temperatures. Increased temperature may have offset other growth/development stressors, such as those related to water chemistry. However, the long-term consequences of these differences on amphibian populations requires further study.

Real-time quantitative PCR assay development and application for assessment of agricultural surface water and various fecal matter for prevalence of Aliarcobacter faecis and Aliarcobacter lanthieri.

BACKGROUND: Aliarcobacter faecis and Aliarcobacter lanthieri are recently identified as emerging human and animal pathogens. In this paper, we demonstrate the development and optimization of two direct DNA-based quantitative real-time PCR assays using species-specific oligonucleotide primer pairs derived from rpoB and gyrA genes for A. faecis and A. lanthieri, respectively. Initially, the specificity of primers and amplicon size of each target reference strain was verified and confirmed by melt curve analysis. Standard curves were developed with a minimum quantification limit of 100 cells mL- 1 or g- 1 obtained using known quantities of spiked A. faecis and A. lanthieri reference strains in autoclaved agricultural surface water and dairy cow manure samples. RESULTS: Each species-specific qPCR assay was validated and applied to determine the rate of prevalence and quantify the total number of cells of each target species in natural surface waters of an agriculturally-dominant and non-agricultural reference watershed. In addition, the prevalence and densities were determined for human and various animal (e.g., dogs, cats, dairy cow, and poultry) fecal samples. Overall, the prevalence of A. faecis for surface water and feces was 21 and 28%, respectively. The maximum A. faecis concentration for water and feces was 2.3 × 107 cells 100 mL- 1 and 1.2 × 107 cells g- 1, respectively. A. lanthieri was detected at a lower frequency (2%) with a maximum concentration in surface water of 4.2 × 105 cells 100 mL- 1; fecal samples had a prevalence and maximum density of 10% and 2.0 × 106 cells g- 1, respectively. CONCLUSIONS: The results indicate that the occurrence of these species in agricultural surface water is potentially due to fecal contamination of water from livestock, human, or wildlife as both species were detected in fecal samples. The new real-time qPCR assays can facilitate rapid and accurate detection in < 3 h to quantify total numbers of A. faecis and A. lanthieri cells present in various complex environmental samples.

Qualifying the effects of single and multiple stressors on the food web structure of Dutch drainage ditches using a literature review and conceptual models.

In September 2017, a workshop was held at Wageningen University and Research to determine the current state of knowledge of multiple stressor effects on aquatic ecosystems and to assess how to improve prediction of these effects. We developed a theoretical framework that integrates species-level responses to stressors to predict how these effects propagate through higher levels of biological organisation. Here, we present the application of the framework for drainage ditch ecosystems in the Netherlands. We used food webs to assess single and multiple stressor effects of common stressors on ditch communities. We reviewed the literature for the effects of targeted stressors (nutrients, pesticides, dredging and mowing, salinization, and siltation) on each functional group present in the food web and qualitatively assessed the relative sensitivity of groups. Using this information, we created a stressor-response matrix of positive and negative direct effects of each stressor-functional group combination. Fungicides, salinization, and sedimentation were identified as particularly detrimental to most groups, although destructive management practices, such as dredging with almost complete community removal, would take precedence depending on frequency. Using the stressor-response matrix we built, first, a series of conceptual null models of single stressor effects on food web structure and, second, a series of additive null models to illustrate potential paired-stressor effects. We compared these additive null models with published studies of the same pairs of combined single stressors to explore more complex interactions. Our approach serves as a first-step to considering multiple stressor scenarios in systems that are understudied or data-poor and as a baseline from which more complex models that include indirect effects and quantitative data may be developed. We make specific suggestions for appropriate management strategies that could be taken to support the biodiversity of these systems for individual stressors and their combined impacts.

Towards a general framework for the assessment of interactive effects of multiple stressors on aquatic ecosystems: Results from the Making Aquatic Ecosystems Great Again (MAEGA) workshop.

A workshop was held in Wageningen, The Netherlands, in September 2017 to collate data and literature on three aquatic ecosystem types (agricultural drainage ditches, urban floodplains, and urban estuaries), and develop a general framework for the assessment of multiple stressors on the structure and functioning of these systems. An assessment framework considering multiple stressors is crucial for our understanding of ecosystem responses within a multiply stressed environment, and to inform appropriate environmental management strategies. The framework consists of two components: (i) problem identification and (ii) impact assessment. Both assessments together proceed through the following steps: 1) ecosystem selection; 2) identification of stressors and quantification of their intensity; 3) identification of receptors or sensitive groups for each stressor; 4) identification of stressor-response relationships and their potential interactions; 5) construction of an ecological model that includes relevant functional groups and endpoints; 6) prediction of impacts of multiple stressors, 7) confirmation of these predictions with experimental and monitoring data, and 8) potential adjustment of the ecological model. Steps 7 and 8 allow the assessment to be adaptive and can be repeated until a satisfactory match between model predictions and experimental and monitoring data has been obtained. This paper is the preface of the MAEGA (Making Aquatic Ecosystems Great Again) special section that includes three associated papers which are also published in this volume, which present applications of the framework for each of the three aquatic systems.