Aging of Copper Nanoparticles in the Marine Environment Regulates Toxicity for a Coastal Phytoplankton Species.

Environmental conditions in aquatic ecosystems transform toxic chemicals over time, influencing their bioavailability and toxicity. Using an environmentally relevant methodology, we tested how exposure to seawater for 1-15 weeks influenced the accumulation and toxicity of copper nanoparticles (nano-Cu) in a marine phytoplankton species. Nano-Cu rapidly agglomerated in seawater and then decreased in size due to Cu dissolution. Dissolution rates declined during weeks 1-4 and remained low until 15 weeks, when the large agglomerates that had formed began to rapidly dissolve again. Marine phytoplankton species were exposed for 5-day periods to nano-Cu aged from 1 to 15 weeks at concentrations from 0.01 to 20 ppm. Toxicity to phytoplankton, measured as change in population growth rate, decreased significantly with particle aging from 0 to 4 weeks but increased substantially in the 15-week treatment due apparently to elevated Cu dissolution of reagglomerated particles. Results indicate that the transformation, fate, and toxicity of nano-Cu in marine ecosystems are influenced by a highly dynamic physicochemical aging process.

Modeled and measured SARS-CoV-2 virus in septic tank systems for wastewater surveillance.

SARS-CoV-2 wastewater surveillance (WWS) at wastewater treatment plants (WWTPs) can reveal sewered community COVID-19 prevalence. For unsewered areas using septic tank systems (STSs) or holding tanks, how to conduct WWS remains unexplored. Here, two large STSs serving Zuma Beach (Malibu, CA) were studied. Supernatant and sludge SARS-CoV-2 concentrations from the directly-sampled STSs parameterized a dynamic solid-liquid separation, mass balance-based model for estimating the infection rate of users. Pumped septage before hauling and upon WWTP disposal was also sampled and assessed. Most (96%) STS sludge samples contained SARS-CoV-2 N1 and N2 genes, with concentrations exceeding the supernatant and increasing with depth while correlating with total suspended solids (TSS). The trucked septage contained N1 and N2 genes which decayed (coefficients: 0.09-0.29 h-1) but remained detectable. Over approximately 5 months starting in December 2020, modeled COVID-19 prevalence estimations among users ranged from 8 to 18%, mirroring a larger metropolitan area for the first 2 months. The approaches herein can inform public health intervention and augment conventional WWS in that: (1) user infection rates for communal holding tanks are estimable and (2) pumped and hauled septage can be assayed to infer where disease is spreading in unsewered areas.

Genetic sequence data evidence that human faecal-associated HF183 sequences are on human skin and in urine.

AIMS: The DNA marker HF183 is a partial 16S rRNA gene sequence highly specific to human-associated Bacteroides including Bacteroides dorei. While HF183 is used to assess human faecal contamination in aquatic environments worldwide, little is known about the existence of HF183 and B. dorei in human microbiomes outside of the human gastrointestinal tract and faeces. METHODS AND RESULTS: Previously published human skin and urine microbiome data sets from five independent human body skin studies, the Human Microbiome Project (HMP) and three independent human urine studies were analysed. The HF183 gene sequence was detected in all skin data sets, with the ratios of positive samples ranging from 0.5% to 36.3%. Popliteal fossa (knee), volar forearm and inguinal (groin) creases were identified as hot spots. HF183 was detected in two of three urine data sets, with ratios of positive samples ranging from 0% to 37.5%. All HF183-containing sequences from these data sets were classified as associated with B. dorei. CONCLUSIONS: HF183 is widespread on human skin and present in urine. SIGNIFICANCE AND IMPACT OF STUDY: Skin and urine microbiomes could be sources of HF183 to environmental waters. Such non-faecal sources of HF183 might explain low concentrations of HF183 in recreational waters when swimmers are present.

SARS-CoV-2 Wastewater Surveillance for Public Health Action.

Wastewater surveillance for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has garnered extensive public attention during the coronavirus disease pandemic as a proposed complement to existing disease surveillance systems. Over the past year, methods for detection and quantification of SARS-CoV-2 viral RNA in untreated sewage have advanced, and concentrations in wastewater have been shown to correlate with trends in reported cases. Despite the promise of wastewater surveillance, for these measurements to translate into useful public health tools, bridging the communication and knowledge gaps between researchers and public health responders is needed. We describe the key uses, barriers, and applicability of SARS-CoV-2 wastewater surveillance for supporting public health decisions and actions, including establishing ethics consideration for monitoring. Although wastewater surveillance to assess community infections is not a new idea, the coronavirus disease pandemic might be the initiating event to make this emerging public health tool a sustainable nationwide surveillance system, provided that these barriers are addressed.

Limited Bacterial Removal in Full-Scale Stormwater Biofilters as Evidenced by Community Sequencing Analysis.

In urban areas, untreated stormwater runoff can pollute downstream surface waters. To intercept and treat runoff, low-impact or "green infrastructure" approaches such as using biofilters are adopted. Yet, actual biofilter pollutant removal is poorly understood; removal is often studied in laboratory columns, with variable removal of viable and culturable microbial cell numbers including pathogens. Here, to assess bacterial pollutant removal in full-scale planted biofilters, stormwater was applied, unspiked or spiked with untreated sewage, in simulated storm events under transient flow conditions, during which biofilter influents versus effluents were compared. Based on microbial biomass, sequences of bacterial community genes encoding 16S rRNA, and gene copies of the human fecal marker HF183 and of the Enterococcus spp. marker Entero1A, removal of bacterial pollutants in biofilters was limited. Dominant bacterial taxa were similar for influent versus effluent aqueous samples within each inflow treatment of either spiked or unspiked stormwater. Bacterial pollutants in soil were gradually washed out, albeit incompletely, during simulated storm flushing events. In post-storm biofilter soil cores, retained influent bacteria were concentrated in the top layers (0-10 cm), indicating that the removal of bacterial pollutants was spatially limited to surface soils. To the extent that plant-associated processes are responsible for this spatial pattern, treatment performance might be enhanced by biofilter designs that maximize influent contact with the rhizosphere.

Physical Properties of Carbon Nanomaterials and Nanoceria Affect Pathways Important to the Nodulation Competitiveness of the Symbiotic N2 -Fixing Bacterium Bradyrhizobium diazoefficiens.

The pathogenicity and antimicrobial properties of engineered nanomaterials (ENMs) are relatively well studied. However, less is known regarding the interactions of ENMs and agriculturally beneficial microorganisms that affect food security. Nanoceria (CeO2 nanoparticles (NPs)), multiwall carbon nanotubes (MWCNTs), graphene nanoplatelets (GNPs), and carbon black (CB) have been previously shown to inhibit symbiotic N2 fixation in soybeans, but direct rhizobial susceptibility is uncertain. Here, Bradyrhizobium diazoefficiens associated with symbiotic N2 fixation in soybeans is assessed, evaluating the role of soybean root exudates (RE) on ENM-bacterial interactions and the effects of CeO2 NPs, MWCNTs, GNPs, and CB on bacterial growth and gene expression. Although bacterial growth is inhibited by 50 mg L-1 CeO2 NPs, MWCNTs, and CB, all ENMs at 0.1 and 10 mg L-1 cause a global transcriptomic response that is mitigated by RE. ENMs may interfere with plant-bacterial signaling, as evidenced by suppressed upregulation of genes induced by RE, and downregulation of genes encoding transport RNA, which facilitates nodulation signaling. MWCNTs and CeO2 NPs inhibit the expression of genes conferring B. diazoefficiens nodulation competitiveness. Surprisingly, the transcriptomic effects on B. diazoefficiens are similar for these two ENMs, indicating that physical, not chemical, ENM properties explain the observed effects.

Effects of carbonaceous nanomaterials on soil-grown soybeans under combined heat and insect stresses.

Because carbonaceous nanomaterials (CNMs) are expected to enter soils, the exposure implications to crop plants and plant-microbe interactions should be understood. Most investigations have been under ideal growth conditions, yet crops commonly experience abiotic and biotic stresses. Little is known how co-exposure to these environmental stresses and CNMs would cause combined effects on plants. We investigated the effects of 1000 mg kg-1 multiwalled carbon nanotubes (CNTs), graphene nanoplatelets (GNPs) and industrial carbon black (CB) on soybeans grown to the bean production stage in soil. Following seed sowing, plants became stressed by heat and infested with an insect (thrips). Consequently, all plants had similarly stunted growth, leaf damage, reduced final biomasses and fewer root nodules compared with healthy control soybeans previously grown without heat and thrips stresses. Thus, CNMs did not significantly influence the growth and yield of stressed soybeans, and the previously reported nodulation inhibition by CNMs was not specifically observed here. However, CNMs did significantly alter two leaf health indicators: the leaf chlorophyll a/b ratio, which was higher in the GNP treatment than in either the control (by 15 %) or CB treatment (by 14 %), and leaf lipid peroxidation, which was elevated in the CNT treatment compared with either the control (by 47 %) or GNP treatment (by 66 %). Overall, these results show that, while severe environmental stresses may impair plant production, CNMs (including CNTs and GNPs) in soil could additionally affect foliar health of an agriculturally important legume.

Carbonaceous Nanomaterials Have Higher Effects on Soybean Rhizosphere Prokaryotic Communities During the Reproductive Growth Phase than During Vegetative Growth.

Carbonaceous nanomaterials (CNMs) can affect agricultural soil prokaryotic communities, but how the effects vary with the crop growth stage is unknown. To investigate this, soybean plants were cultivated in soils amended with 0, 0.1, 100, or 1000 mg kg-1 of carbon black, multiwalled carbon nanotubes (MWCNTs), or graphene. Soil prokaryotic communities were analyzed by Illumina sequencing at day 0 and at the soybean vegetative and reproductive stages. The sequencing data were functionally annotated using the functional annotation of prokaryotic taxa (FAPROTAX) database. The prokaryotic communities were unaffected at day 0 and were altered at the plant vegetative stage only by 0.1 mg kg-1 MWCNTs. However, at the reproductive stage, when pods were filling, most treatments (except 1000 mg kg-1 MWCNTs) altered the prokaryotic community composition, including functional groups associated with C, N, and S cycling. The lower doses of CNMs, which were previously shown to be less agglomerated and thus more bioavailable in soil relative to the higher doses, were more effective toward both overall communities and individual functional groups. Taken together, prokaryotic communities in the soybean rhizosphere can be significantly phylogenetically and functionally altered in response to bioavailable CNMs, especially when soybean plants are actively directing resources to seed production.

Host-Symbiont Interaction Model Explains Non-monotonic Response of Soybean Growth and Seed Production to Nano-CeO2 Exposure.

Recent nanotoxicity studies have demonstrated non-monotonic dose-response mechanisms for planted soybean that have a symbiotic relationship with bacteroids in their root nodules: reduction of growth and seed production was greater for low, as compared to high, exposures. To investigate mechanistic underpinnings of the observed patterns, we formulated an energy budget model coupled to a toxicokinetic module describing bioaccumulation, and two toxicodynamic modules describing toxic effects on host plant and symbionts. By fitting data on plants exposed to engineered CeO2 nanoparticles to the newly formulated model, we show that the non-monotonic patterns can be explained as the interaction of two, individually monotonic, dose-response processes: one for the plant and the other for the symbiont. We further validate the newly formulated model by showing that, without the need for additional parameters, the model successfully predicts changes in dinitrogen fixation potential as a function of exposure (dinitrogen fixation potential data not used in model fitting). The symbiont buffers overall toxicity only when, in the absence of exposure to a toxicant, it has a parasitic interaction with the host plant. If the interaction is mutualistic or commensal, there is no buffering and only monotonic toxic responses are possible. Because the model is based on general biological principles, we expect it to be applicable to other similar symbiotic systems, especially other nodule-forming legumes.

Spatial Models of Sewer Pipe Leakage Predict the Occurrence of Wastewater Indicators in Shallow Urban Groundwater.

Twentieth century municipal wastewater infrastructure greatly improved U.S. urban public health and water quality. However, sewer pipes deteriorate, and their accumulated structural defects may release untreated wastewater to the environment via acute breaks or insidious exfiltration. Exfiltrated wastewater constitutes a loss of potentially reusable water and delivers a complex and variable mix of contaminants to urban shallow groundwater. Yet, predicting where deteriorated sewers impinge on shallow groundwater has been challenging. Here we develop and test a spatially explicit model of exfiltration probability based on pipe attributes and groundwater elevation without prior knowledge of exfiltrating defect locations. We find that models of exfiltration probability can predict the probable occurrence in underlying shallow groundwater of established wastewater indicators including the artificial sweetener acesulfame, tryptophan-like fluorescent dissolved organic matter, nitrate, and a stable isotope of water (δ18O). The strength of the association between exfiltration probability and indicators of wastewater increased when multiple pipe attributes, distance weighting, and groundwater flow direction were considered in the model. The results prove that available sanitary sewer databases and groundwater digital elevation data can be analyzed to predict where pipes are likely leaking and contaminating groundwater. Such understanding could direct sewer infrastructure reinvestment toward water resource protection.

Bioaccumulation of Multiwall Carbon Nanotubes in Tetrahymena thermophila by Direct Feeding or Trophic Transfer.

Consumer goods contain multiwall carbon nanotubes (MWCNTs) that could be released during product life cycles into the environment, where their effects are uncertain. Here, we assessed MWCNT bioaccumulation in the protozoan Tetrahymena thermophila via trophic transfer from bacterial prey (Pseudomonas aeruginosa) versus direct uptake from growth media. The experiments were conducted using (14)C-labeled MWCNT ((14)C-MWCNT) doses at or below 1 mg/L, which proved subtoxic since there were no adverse effects on the growth of the test organisms. A novel contribution of this study was the demonstration of the ability to quantify MWCNT bioaccumulation at low (sub µg/kg) concentrations accomplished by employing accelerator mass spectrometry (AMS). After the treatments with MWCNTs at nominal concentrations of 0.01 mg/L and 1 mg/L, P. aeruginosa adsorbed considerable amounts of MWCNTs: (0.18 ± 0.04) µg/mg and (21.9 ± 4.2) µg/mg bacterial dry mass, respectively. At the administered MWCNT dose of 0.3 mg/L, T. thermophila accumulated up to (0.86 ± 0.3) µg/mg and (3.4 ± 1.1) µg/mg dry mass by trophic transfer and direct uptake, respectively. Although MWCNTs did not biomagnify in the microbial food chain, MWCNTs bioaccumulated in the protozoan populations regardless of the feeding regime, which could make MWCNTs bioavailable for organisms at higher trophic levels.

Long-Term Effects of Multiwalled Carbon Nanotubes and Graphene on Microbial Communities in Dry Soil.

Little is known about the long-term effects of engineered carbonaceous nanomaterials (ECNMs) on soil microbial communities, especially when compared to possible effects of natural or industrial carbonaceous materials. To address these issues, we exposed dry grassland soil for 1 year to 1 mg g(-1) of either natural nanostructured material (biochar), industrial carbon black, three types of multiwalled carbon nanotubes (MWCNTs), or graphene. Soil microbial biomass was assessed by substrate induced respiration and by extractable DNA. Bacterial and fungal communities were examined by terminal restriction fragment length polymorphism (T-RFLP). Microbial activity was assessed by soil basal respiration. At day 0, there was no treatment effect on soil DNA or T-RFLP profiles, indicating negligible interference between the amended materials and the methods for DNA extraction, quantification, and community analysis. After a 1-year exposure, compared to the no amendment control, some treatments reduced soil DNA (e.g., biochar, all three MWCNT types, and graphene; P < 0.05) and altered bacterial communities (e.g., biochar, carbon black, narrow MWCNTs, and graphene); however, there were no significant differences across the amended treatments. These findings suggest that ECNMs may moderately affect dry soil microbial communities but that the effects are similar to those from natural and industrial carbonaceous materials, even after 1-year exposure.

Microfiber Masses Recovered from Conventional Machine Washing of New or Aged Garments.

Synthetic textiles can shed numerous microfibers during conventional washing, but evaluating environmental consequences as well as source-control strategies requires understanding mass releases. Polyester apparel accounts for a large proportion of the polyester market, and synthetic jackets represent the broadest range in apparel construction, allowing for potential changes in manufacturing as a mitigation measure to reduce microfiber release during laundering. Here, detergent-free washing experiments were conducted and replicated in both front- and top-load conventional home machines for five new and mechanically aged jackets or sweaters: four from one name-brand clothing manufacturer (three majority polyester fleece, and one nylon shell with nonwoven polyester insulation) and one off-brand (100% polyester fleece). Wash water was filtered to recover two size fractions (>333 µm and between 20 and 333 µm); filters were then imaged, and microfiber masses were calculated. Across all treatments, the recovered microfiber mass per garment ranged from approximately 0 to 2 g, or exceeding 0.3% of the unwashed garment mass. Microfiber masses from top-load machines were approximately 7 times those from front-load machines; garments mechanically aged via a 24 h continuous wash had increased mass release under the same wash protocol as new garments. When published wastewater treatment plant influent characterization and microfiber removal studies are considered, washing synthetic jackets or sweaters as per this study would account for most microfibers entering the environment.

Considerations of Environmentally Relevant Test Conditions for Improved Evaluation of Ecological Hazards of Engineered Nanomaterials.

Engineered nanomaterials (ENMs) are increasingly entering the environment with uncertain consequences including potential ecological effects. Various research communities view differently whether ecotoxicological testing of ENMs should be conducted using environmentally relevant concentrations-where observing outcomes is difficult-versus higher ENM doses, where responses are observable. What exposure conditions are typically used in assessing ENM hazards to populations? What conditions are used to test ecosystem-scale hazards? What is known regarding actual ENMs in the environment, via measurements or modeling simulations? How should exposure conditions, ENM transformation, dose, and body burden be used in interpreting biological and computational findings for assessing risks? These questions were addressed in the context of this critical review. As a result, three main recommendations emerged. First, researchers should improve ecotoxicology of ENMs by choosing test end points, duration, and study conditions-including ENM test concentrations-that align with realistic exposure scenarios. Second, testing should proceed via tiers with iterative feedback that informs experiments at other levels of biological organization. Finally, environmental realism in ENM hazard assessments should involve greater coordination among ENM quantitative analysts, exposure modelers, and ecotoxicologists, across government, industry, and academia.

Toxicity of metal oxide nanoparticles in Escherichia coli correlates with conduction band and hydration energies.

Metal oxide nanoparticles (MOx NPs) are used for a host of applications, such as electronics, cosmetics, construction, and medicine, and as a result, the safety of these materials to humans and the environment is of considerable interest. A prior study of 24 MOx NPs in mammalian cells revealed that some of these materials show hazard potential. Here, we report the growth inhibitory effects of the same series of MOx NPs in the bacterium Escherichia coli and show that toxicity trends observed in E. coli parallel those seen previously in mammalian cells. Of the 24 materials studied, only ZnO, CuO, CoO, Mn2O3, Co3O4, Ni2O3, and Cr2O3 were found to exert significant growth inhibitory effects; these effects were found to relate to membrane damage and oxidative stress responses in minimal trophic media. A correlation of the toxicological data with physicochemical parameters of MOx NPs revealed that the probability of a MOx NP being toxic increases as the hydration enthalpy becomes less negative and as the conduction band energy approaches those of biological molecules. These observations are consistent with prior results observed in mammalian cells, revealing that mechanisms of toxicity of MOx NPs are consistent across two very different taxa. These results suggest that studying nanotoxicity in E. coli may help to predict toxicity patterns in higher organisms.

Soybean susceptibility to manufactured nanomaterials with evidence for food quality and soil fertility interruption.

Based on previously published hydroponic plant, planktonic bacterial, and soil microbial community research, manufactured nanomaterial (MNM) environmental buildup could profoundly alter soil-based food crop quality and yield. However, thus far, no single study has at once examined the full implications, as no studies have involved growing plants to full maturity in MNM-contaminated field soil. We have done so for soybean, a major global commodity crop, using farm soil amended with two high-production metal oxide MNMs (nano-CeO(2) and -ZnO). The results provide a clear, but unfortunate, view of what could arise over the long term: (i) for nano-ZnO, component metal was taken up and distributed throughout edible plant tissues; (ii) for nano-CeO(2), plant growth and yield diminished, but also (iii) nitrogen fixation--a major ecosystem service of leguminous crops--was shut down at high nano-CeO(2) concentration. Juxtaposed against widespread land application of wastewater treatment biosolids to food crops, these findings forewarn of agriculturally associated human and environmental risks from the accelerating use of MNMs.

Ecological nanotoxicology: integrating nanomaterial hazard considerations across the subcellular, population, community, and ecosystems levels.

Research into the health and environmental safety of nanotechnology has seriously lagged behind its emergence in industry. While humans have often adopted synthetic chemicals without considering ancillary consequences, the lessons learned from worldwide pollution should motivate making nanotechnology compatible with environmental concerns. Researchers and policymakers need to understand exposure and harm of engineered nanomaterials (ENMs), currently nanotechnology's main products, to influence the ENM industry toward sustainable growth. Yet, how should research proceed? Standard toxicity testing anchored in single-organism, dose-response characterizations does not adequately represent real-world exposure and receptor scenarios and their complexities. Our approach is different: it derives from ecology, the study of organisms' interactions with each other and their environments. Our approach involves the characterization of ENMs and the mechanistic assessment of their property-based effects. Using high throughput/content screening (HTS/HCS) with cells or environmentally-relevant organisms, we measure the effects of ENMs on a subcellular or population level. We then relate those effects to mechanisms within dynamic energy budget (DEB) models of growth and reproduction. We reconcile DEB model predictions with experimental data on organism and population responses. Finally, we use microcosm studies to measure the potential for community- or ecosystem-level effects by ENMs that are likely to be produced in large quantities and for which either HTS/HCS or DEB modeling suggest their potential to harm populations and ecosystems. Our approach accounts for ecological interactions across scales, from within organisms to whole ecosystems. Organismal ENM effects, if propagated through populations, can alter communities comprising multiple populations (e.g., plant, fish, bacteria) within food webs. Altered communities can change ecosystem services: processes that cycle carbon, nutrients, and energy, and regulate Earth's waters and atmosphere. We have shown ENM effects on populations, communities, and ecosystems, including transfer and concentration of ENMs through food chains, for a range of exposure scenarios; in many cases, we have identified subcellular ENM effects mechanisms. To keep pace with ENM development, rapid assessment of the mechanisms of ENM effects and modeling are needed. DEB models provide a method for mathematically representing effects such as the generation of reactive oxygen species and their associated damage. These models account for organism-level effects on metabolism and reproduction and can predict outcomes of ENM-organism combinations on populations; those predictions can then suggest ENM characteristics to be avoided. HTS/HCS provides a rapid assessment tool of the ENM chemical characteristics that affect biological systems; such results guide and expand DEB model expressions of hazard. Our approach addresses ecological processes in both natural and managed ecosystems (agriculture) and has the potential to deliver timely and meaningful understanding towards environmentally sustainable nanotechnology.

Integrated approach to evaluating the toxicity of novel cysteine-capped silver nanoparticles to Escherichia coli and Pseudomonas aeruginosa.

Because of microbial resistance to conventional antibiotics, there is increasing interest in silver, including silver nanoparticles (nano-Ag), in antimicrobial applications. However, questions remain regarding the relative roles of nano-Ag particles, versus Ag(+) ions released from nano-Ag dissolution, in imparting bacterial toxicity. Here, we developed a novel nano-Ag that, based on its cysteine cap, was expected to dissolve slowly and thus potentially allow for differentiating nanoparticle, versus ionic, effects of Ag. The nano-Ag was systematically tested for its differential toxicity to Escherichia coli and Pseudomonas aeruginosa. Bacterial growth, reactive oxygen species (ROS) generation, particle dissolution, cellular electron transfer activity, and cell membrane damage and potential were evaluated. In minimal growth medium, E. coli and P. aeruginosa growth were slowed at 100 mg L(-1) (0.93 mM) and 5 mg L(-1) (0.046 mM), respectively; P. aeruginosa was completely inhibited at and above 10 mg L(-1) (0.093 mM). For both strains, toxicity was associated with ROS and cell membrane damage. Based on comparisons to AgNO3 exposures, toxicity from nano-Ag was due to Ag(+) ions and not intact nano-Ag, even though nanoparticle dissolution was less than 2% in minimal growth medium. Because of their stability and slow Ag(+) ion release, the cysteine-capped nano-Ag particles here are useful to antimicrobial applications. Additionally, our systematic approach to evaluating toxicity, membrane damage, and ROS generation can be applied with other nanomaterials and bacteria.

Microbial source tracking in a coastal California watershed reveals canines as controllable sources of fecal contamination.

Elevated levels of fecal indicator bacteria (FIB), including Escherichia coli and enterococci, trigger coastal beach advisories and signal public health risks. Solving FIB pollution in suburban coastal watersheds is challenging, as there are many potential sources. The Arroyo Burro watershed in Santa Barbara, CA is an example, with its popular, but chronically FIB-contaminated beach. To address, a microbial source tracking study was performed. Surface waters were sampled over 2 years, FIB were quantified, and DNA was analyzed for host-associated fecal markers. Surf zone FIB were only elevated when the coastal lagoon was discharging. Among the fecal sources into the lagoon, including upstream human sources and coastal birds, canines were the most important. Canine sources included input via upstream creek water, which decreased after creek-side residences were educated about proper pet waste disposal, and direct inputs to the lagoon and surf zone, where dog waste could have been tidally exchanged with the lagoon. Based on this study, canine waste can be an influential, yet controllable, fecal source to suburban coastal beaches.