Fate and effects of amphoteric surfactants in the aquatic environment

Amphoteric surfactants form part of specialty surfactants available for formulators to improve or design new formulations in response to environmental, toxicity, safety and performance demands. Nevertheless, limited information on the ecological properties of amphoterics is available. In the present work, the aerobic and anaerobic biodegradability and the aquatic toxicity of different types of amphoteric surfactants (three alkyl betaines, one alkylamido betaine and three alkyl imidazoline derivatives) were studied. The amphoteric surfactants tested were readily biodegradable under aerobic conditions (CO(2) headspace test) and alkylamido betaines and alkyl imidazoline derivatives were also easily biodegradable under anaerobic conditions (test based on the ECETOC method). Toxicity to Photobacterium phosphoreum and Daphnia magna increased with the fatty chain length of the surfactant. The EC(50) toxicity values of the amphoterics tested were higher than 5 mg/L, and alkyl imidazoline derivatives, with EC(50) values from 20 to >200 mg/L, showed the lowest aquatic toxicity.     

Toxicological effects of disinfections using sodium hypochlorite on aquatic organisms and its contribution to AOX formation in hospital wastewater

Sodium hypochlorite (NaOCl) is often used for disinfecting hospital wastewater in order to prevent the spread of pathogenic microorganisms, causal agents of nosocomial infectious diseases. Chlorine disinfectants in wastewater react with organic matters, giving rise to organic chlorine compounds such as AOX (halogenated organic compounds adsorbable on activated carbon), which are toxic for aquatic organisms and are persistent environmental contaminants. The aim of this study was to evaluate the toxicity on aquatic organisms of hospital wastewater from services using NaOCl in pre-chlorination. Wastewater samples from the infectious and tropical diseases department of a hospital of a large city in southeast of France were collected. Three samples per day were collected in the connecting well department at 9 a.m., 1 p.m. and 5 p.m. during 8 days from 13 March to 22 March 2001, and a mixture was made at 6 p.m. with the three samples in order to obtain a representative sample for the day. The toxicity test comprised the 24-h EC50 on Daphnia magna and a bioluminescence assay using Vibrio fischeri photobacteria. Fecal coliforms and physicochemical analyses such as total organic carbon (TOC), chloride, AOX, total suspended solids (TSS) and chemical oxygen demand (COD) were carried out. Wastewater samples highlighted considerable acute toxicity on D. magna and V. fischeri photobacteria. However, low most probable numbers (MPN), ranging from <3 to 2400 for 100 ml, were detected for fecal coliforms. Statistical analysis, with a confidence interval of 95%, gave a strong linear regression assessed with r=0.98 between AOX concentrations and EC50 (TU) on daphnia. The identification of an ideal concentration of NaOCl in disinfecting hospital wastewater, i.e. its non-observed effect concentration (NOEC) on algae and D. magna, seems to be a research issue that could facilitate the control of AOX toxicity effects on aquatic organisms. Therefore, it would be necessary to monitor the biocide properties of NaOCl on fecal coliforms at various doses and its toxicity effects on aquatic organisms.     

Fate, behavior and effects of surfactants and their degradation products in the environment

Surfactants are widely used in household and industrial products. After use, surfactants as well as their products are mainly discharged into sewage treatment plants and then dispersed into the environment through effluent discharge into surface waters and sludge disposal on lands. Surfactants have different behavior and fate in the environment. Nonionic and cationic surfactants had much higher sorption on soil and sediment than anionic surfactants such as LAS. Most surfactants can be degraded by microbes in the environment although some surfactants such as LAS and DTDMAC as well as alkylphenols may be persistent under anaerobic conditions. LAS were found to degrade in sludge amended soils with a half-lives of 7 to 33 days. Most surfactants are not acutely toxic to organisms at environmental concentrations and aquatic chronic toxicity of surfactants occurred at concentrations usually greater than 0.1 mg/L. However, alkylphenols have shown to be capable of inducing the production of vitellogenin in male fish at a concentration as low as 5 microg/L. More toxicity data are needed to assess the effects on terrestrial organisms such as plants.     

Review of pollutants in petroleum refinery wastewaters and effect upon aquatic organisms

In the past decade, the number of organic chemical contaminants identified in oil refinery wastewaters has risen from less than 20 to over 300. This increase in knowledge was made possible by advances in gas chromatography-mass spectrometry instrumentation and new techniques to maximize resolution of capillary chromatography. The types of compounds identified in refinery wastewaters are similar to those identified in crude oils; i.e., aliphatic, aromatic, alkyl aromatic, polynuclear aromatic hydrocarbons, and some hydrocarbons containing nitrogen, sulfur, and oxygen. The contaminants in oil refinery wastewaters acutely lethal to aquatic organisms can be removed by biological treatment systems. However, biological treatment systems may not be capable of removing all deleterious contaminants. Long-term fathead minnow bioassays of biologically treated refinery wastewaters resulted in 10%–50% mortality after 14–16 days of exposure. The lethal effects were eliminated by sequential treatment with dual media filtration-activated carbon adsorption.     

Perfluoroalkyl compounds in Danish wastewater treatment plants and aquatic environments

This study reports the results of a screening survey of perfluoroalkyl compounds (PFCs) in the Danish environment. The study included point sources (municipal and industrial wastewater treatment plants and landfill sites) and the marine and freshwater environments. Effluent and influent water and sewage sludge were analysed for point sources. Sediment, blue mussels (Mytilus edulis) and liver from plaice (Pleuronectes platessa), flounder (Platichthys flesus) and eel (Anguilla anguilla) were analysed for the freshwater and marine environments. The results obtained show a diffuse PFCs contamination of the Danish environment with concentrations similar to those measured in other countries with the absence of primary contamination sources such as fluorochemical production. PFOS and PFOA were generally the most dominating PFCs measured in both point sources and the aquatic environments. PFCs were found in both inflow and outflow water and sewage sludge from municipal and industrial wastewater treatment plants (WWTPs), indicating that WWTPs can be significant sources to PFCs in the environment. This is also reflected in the locally elevated PFCs concentrations found in fish like eels from shallow freshwater and marine areas. However, the highest PFCs concentrations found in fish in this study was in plaice from the Skagerrak (156 ng/g wet weight PFOS), but it is unknown if this can be related to significant sources in the North Sea region or to differences between species. The concentrations of PFCs were below the detection limit in all analysed freshwater and marine samples of sediment and mussels. Despite the relatively low PFCs concentrations measured in marine fish, the high bioaccumulation potential of PFCs, particularly PFOS, may lead to high concentrations of PFCs in marine mammals as shown by previous investigations.     

Polycyclic aromatic hydrocarbons and effects on marine organisms in the Istanbul Strait

Surficial sediments and mussels (Mytilus galloprovincialis) from the Istanbul Strait and Marmara Sea were analysed for sixteen parent polycyclic aromatic hydrocarbon (PAH) contents by gas chromatography-mass spectrometry (GC-MS) employing isotope dilution technique. Microalgae toxicity testing was applied to sediment elutriates and biological responses in terms of filtration rate and lysosomal stability were measured in mussels. Total PAH concentrations ranged from 2.1 to 3152 ng g^? 1 dry wt in sediments and from 43–601 ng g^? 1 wet weight in mussels. Molecular indices of phenanthrene/ anthracene, fluoranthene/pyrene and benzo(a)anthracene/chrysene were used to differentiate between pyrolytic and petroleum origin. Results showed that most of the contamination originates from high temperature pyrolytic inputs with some slight contribution of petrogenic PAH. PAH in sediments were frequently lower than the National Oceanic and Atmospheric Administration (NOAA)–ERM (Effects Range Medium) index. Results of sediment elutriate toxicity testing and biomarkers indicate that the cause of negative effects in sediments may result from different classes of pollutants and does not only relate with PAH contamination. Mussels from most of the stations showed both reduced lysosomal membrane stability and filtration rate indicating disturbed health although the two biomarker results did not always complement each other. The effect studies showed that the pollutants in the strait ecosystem have more pronounced effects in the middle parts than those at the Black Sea entrance.     

Community effects of carbon nanotubes in aquatic sediments

Aquatic sediments form an important sink for manufactured nanomaterials, like carbon nanotubes (CNT) and fullerenes, thus potentially causing adverse effects to the aquatic environment, especially to benthic organisms. To date, most nanoparticle effect studies used single species tests in the laboratory, which lacks ecological realism. Here, we studied the effects of multiwalled CNT (MWCNT) contaminated sediments on benthic macroinvertebrate communities. Sediment was taken from an unpolluted site, cleaned from invertebrates, mixed with increasing levels of MWCNTs (0, 0.002, 0.02, 0.2 and 2g/kg dry weight), transferred to trays and randomly relocated in the original unpolluted site, which now acted as a donor system for recolonization by benthic species. After three months of exposure, the trays were regained, organic (OC) and residual carbon (RC) were measured, and benthic organisms and aquatic macrophytes were identified. ANOVA revealed a significantly higher number of individuals with increasing MWCNT concentrations. The Shannon index showed no significant effect of MWCNT addition on biodiversity. Multivariate statistics applied to the complete macroinvertebrate dataset, did show effects on the community level. Principal Component Analysis (PCA) showed differences in taxa composition related to MWCNT levels indicating differences in sensitivity of the taxa. Redundancy Analysis (RDA) revealed that MWCNT dose, presence of macrophytes, and spatial distribution explained 38.3% of the total variation in the data set, of which MWCNT dose contributed with 18.9%. Still, the net contribution of MWCNT dose was not statistically significant, indicating that negative community effects are not likely to occur at environmentally relevant future CNT concentrations in aquatic sediments.     

The effects of atrazine on dissolved oxygen and nitrate concentrations in aquatic systems

The effects of the herbicide atrazine, on nitrate-nitrogen (nitrate-N) and dissolved oxygen concentrations in aquatic microcosms were determined. Treatment levels consisted of a control, 50, 100, and 150 μg/1 atrazine with four replicates per treatment. From a stock atrazine solution serial dilutions were prepared for each treatment level. The serial dilutions were added as single daily doses in 150 mL increments to the appropriate treatment. Dissolved oxygen was measured twice daily for each ssytem. Nitrate-N concentrations were measured twice weekly for the duration of the experiment.Dissolved oxygen was not resistant to the atrazine perturbation and decreased to concentrations approximating 0.0 mg/L for all atrazine concentrations. However, dissolved oxygen concentrations were resilient to perturbation and recovered to pretreatment levels within 48 h following cessation of toxicant input. Nitrate-N concentrations also were not resistant to the atrazine perturbation and decreased to more than 90% of the pretreatment concentrations following atrazine introduction. Nitrate-N concentrations for atrazine treatments exhibited high resilience to the perturbation. Nitrate-N concentrations recovered to approximately 90% of the preperturbation nitrate-N concentrations 48 h after cessation of the toxicant.Although small amounts of atrazine may have a significant impact upon aquatic systems by reducing levels of dissolved oxygen and nitrate, there appears to be no permanent ecosystem damage since these systems recover rapidly when atrazine levels return to zero.     

Silver nanoparticles: behaviour and effects in the aquatic environment

This review summarises and evaluates the present knowledge on the behaviour, the biological effects and the routes of uptake of silver nanoparticles (Ag NPs) to organisms, with considerations on the nanoparticle physicochemistry in the ecotoxicity testing systems used. Different types of Ag NP syntheses, characterisation techniques and predicted current and future concentrations in the environment are also outlined. Rapid progress in this area has been made over the last few years, but there is still a critical lack of understanding of the need for characterisation and synthesis in environmental and ecotoxicological studies. Concentration and form of nanomaterials in the environment are difficult to quantify and methodological progress is needed, although sophisticated exposure models show that predicted environmental concentrations (PECs) for Ag NPs in different environmental compartments are at the range of ng L(-1) to mg kg(-1). The ecotoxicological literature shows that concentrations of Ag NPs below the current and future PECs, as low as just a few ng L(-1), can affect prokaryotes, invertebrates and fish indicating a significant potential, though poorly characterised, risk to the environment. Mechanisms of toxicity are still poorly understood although it seems clear that in some cases nanoscale specific properties may cause biouptake and toxicity over and above that caused by the dissolved Ag ion. This review concludes with a set of recommendations for the advancement of understanding of the role of nanoscale silver in environmental and ecotoxicological research.     

Factors affecting metal toxicity to (and accumulation by) aquatic organisms — Overview

This literature review encompasses aquatic environmental toxicities of metals and metalloids. The emphasis is on the influencing factors on metal toxicity to aquatic organisms. The effects of environmental factors on metal uptake are also discussed. The factors can be divided into biotic and abiotic. The biotic factors include tolerance, size and life stages, species, and nutrition related to the test organisms. The abiotic factors include organic substances, pH, temperature, alkalinity and hardness, inorganic ligands, interactions, sediments, and others. These factors can alter metal toxicity in the aquatic environment substantially, mostly causing attenuating effect. The literature shows divergent results. For example, the interactions between Cd and Zn were reported to be synergistic by some researchers and antagonistic by others. It is recommended that environmental hazard assessment takes into consideration the results of standard toxicity tests and site-specific conditions which can moderate metal toxicity considerably.     

Determination of pharmaceutical compounds in hospital effluents and their contribution to wastewater treatment works

A method was developed for the simultaneous determination of almost 40 pharmaceuticals; including antidepressants, non-steroidal anti-inflammatories, analgesics, hypolipidemics, α- and β-blockers, an anti cancer drug, anti-fungal agents, an opiate, an antibiotic, an anti-coagulant, a diuretic, an anti-anginal and an anti-diabetic compound. This was used to assess the contribution of pharmaceuticals originating from hospital effluents to one of Oslo city's wastewater treatment works. Some pharmaceuticals were found to contribute to more of the wastewater loading than others. 11% of the propranolol entering the wastewater treatment works stems from hospital effluent, approximately 2% of the atenolol, carbemazepine, metaprolol and atorvastatin, and for several other compounds the contribution is less than 1%.This assessment shows that point sources discharges from hospitals typically make a small contribution to the overall pharmaceutical load when compared to municipal areas, however this varies from substance to substance and is not the case when a drug's use is primarily hospital based.     

Occurrence,fate and effects of azoxystrobin in aquatic ecosystems: A review

The use of pesticides for crop protection may result in the presence of toxic residues in environmental matrices. In the aquatic environment, pesticides might freely dissolve in the water or bind to suspended matter and to the sediments, and might be transferred to the organisms' tissues during bioaccumulation processes, resulting in adverse consequences to non-target species. One such group of synthetic organic pesticides widely used worldwide to combat pathogenic fungi affecting plants is the strobilurin chemical group. Whereas they are designed to control fungal pathogens, their general modes of action are not specific to fungi. Consequently, they can be potentially toxic to a wide range of non-target organisms. The present work had the intent to conduct an extensive literature review to find relevant research on the occurrence, fate and effects of azoxystrobin, the first patent of the strobilurin compounds, in aquatic ecosystems in order to identify strengths and gaps in the scientific database. Analytical procedures and existing legislation and regulations were also assessed. Data gathered in the present review revealed that analytical reference standards for the most relevant environmental metabolites of azoxystrobin are needed. Validated confirmatory methods for complex matrices, like sediment and aquatic organisms' tissues, are very limited. Important knowledge of base-line values of azoxystrobin and its metabolites in natural tropical and estuarine/marine ecosystems is lacking. Moreover, some environmental concentrations of azoxystrobin found in the present review are above the Regulatory Acceptable Concentration (RAC) in what concerns risk to aquatic invertebrates and the No Observed Ecologically Adverse Effect Concentration (NOEAEC) reported for freshwater communities. The present review also showed that there are very few data on azoxystrobin toxicity to different aquatic organisms, especially in what concerns estuarine/marine organisms. Besides, toxicity studies mostly address azoxystrobin and usually neglect the more relevant environmental metabolites. Further work is also required in what concerns effects of exposure to multi-stressors, e.g. pesticide mixtures. Even though Log K_ow for azoxystrobin and R234886, the main metabolite of azoxystrobin in water, are below 3, the bio-concentration factor and the bioaccumulation potential for azoxystrobin are absent in the literature. Moreover, no single study on bioaccumulation and biomagnification processes was found in the present review.     

Ultraviolet irradiation of municipal wastewater: Evaluation of effects on organic constituents

A comparison of chromatograms derived from the UV-absorbing and oxidizable constituents present in primary and secondary municipal wastewater effluents indicates that exposure to UV irradiation at disinfection levels results in only slight chemical changes. The most pronounced chemical effects to nonvolatile organic constituents in wastewater effluents have been observed at irradiation levels in excess of those necessary for disinfection. Aliquots of effluents before and after exposure to varying levels of UV irradiation were concentrated by lyophilization prior to liquid chromatographic (LC) analysis. Anion-exchange chromatographic techniques utilizing a modified ultraviolet detector coupled in series with a cerate oxidative monitor provided excellent resolution and sensitivity in the determination of chemical changes occurring in the effluents as a result of exposure to different irradiation levels. The total coliform counts for each irradiated effluent were determined by the membrane-filter technique to evaluate the effectiveness of UV irradiation as a disinfection process for municipal wastewaters. Changes in biochemical oxygen demand (BOD), chemical oxygen demand (COD), total organic carbon (TOC), and total Kjeldahl nitrogen (TKN) at different levels of exposure to UV irradiation are presented. The results of these determinations and the gas chromatography/mass spectrometry data characterizing the observed chemical changes are discussed.     

Ecological effects of various toxic agents on the aquatic microcosm in comparison with acute ionizing radiation

The purpose of this study was an evaluation of the effect levels of various toxic agents compared with acute doses of ionizing radiation for the experimental model ecosystem, i.e., microcosm mimicking aquatic microbial communities. For this purpose, the authors used the microcosm consisting of populations of the flagellate alga Euglena gracilis as a producer, the ciliate protozoan Tetrahymena thermophila as a consumer and the bacterium Escherichia coli as a decomposer. Effects of aluminum and copper on the microcosm were investigated in this study, while effects of gamma-rays, ultraviolet radiation, acidification, manganese, nickel and gadolinium were reported in previous studies. The microcosm could detect not only the direct effects of these agents but also the community-level effects due to the interspecies interactions or the interactions between organisms and toxic agents. The authors evaluated doses or concentrations of each toxic agent which had the following effects on the microcosm: (1) no effects; (2) recognizable effects, i.e., decrease or increase in the cell densities of at least one species; (3) severe effects, i.e., extinction of one or two species; and (4) destructive effects, i.e., extinction of all species. The resulting effects data will contribute to an ecological risk assessment of the toxic agents compared with acute doses of ionizing radiation.     

Ecological and toxicological effects of inorganic nitrogen pollution in aquatic ecosystems: A global assessment

We provide a global assessment, with detailed multi-scale data, of the ecological and toxicological effects generated by inorganic nitrogen pollution in aquatic ecosystems. Our synthesis of the published scientific literature shows three major environmental problems: (1) it can increase the concentration of hydrogen ions in freshwater ecosystems without much acid-neutralizing capacity, resulting in acidification of those systems; (2) it can stimulate or enhance the development, maintenance and proliferation of primary producers, resulting in eutrophication of aquatic ecosystems; (3) it can reach toxic levels that impair the ability of aquatic animals to survive, grow and reproduce. Inorganic nitrogen pollution of ground and surface waters can also induce adverse effects on human health and economy. Because reductions in SO2 emissions have reduced the atmospheric deposition of H2SO4 across large portions of North America and Europe, while emissions of NOx have gone unchecked, HNO3 is now playing an increasing role in the acidification of freshwater ecosystems. This acidification process has caused several adverse effects on primary and secondary producers, with significant biotic impoverishments, particularly concerning invertebrates and fishes, in many atmospherically acidified lakes and streams. The cultural eutrophication of freshwater, estuarine, and coastal marine ecosystems can cause ecological and toxicological effects that are either directly or indirectly related to the proliferation of primary producers. Extensive kills of both invertebrates and fishes are probably the most dramatic manifestation of hypoxia (or anoxia) in eutrophic and hypereutrophic aquatic ecosystems with low water turnover rates. The decline in dissolved oxygen concentrations can also promote the formation of reduced compounds, such as hydrogen sulphide, resulting in higher adverse (toxic) effects on aquatic animals. Additionally, the occurrence of toxic algae can significantly contribute to the extensive kills of aquatic animals. Cyanobacteria, dinoflagellates and diatoms appear to be major responsible that may be stimulated by inorganic nitrogen pollution. Among the different inorganic nitrogenous compounds (NH4+, NH3, NO2-, HNO2NO3-) that aquatic animals can take up directly from the ambient water, unionized ammonia is the most toxic, while ammonium and nitrate ions are the least toxic. In general, seawater animals seem to be more tolerant to the toxicity of inorganic nitrogenous compounds than freshwater animals, probably because of the ameliorating effect of water salinity (sodium, chloride, calcium and other ions) on the tolerance of aquatic animals. Ingested nitrites and nitrates from polluted drinking waters can induce methemoglobinemia in humans, particularly in young infants, by blocking the oxygen-carrying capacity of hemoglobin. Ingested nitrites and nitrates also have a potential role in developing cancers of the digestive tract through their contribution to the formation of nitrosamines. In addition, some scientific evidences suggest that ingested nitrites and nitrates might result in mutagenicity, teratogenicity and birth defects, contribute to the risks of non-Hodgkin's lymphoma and bladder and ovarian cancers, play a role in the etiology of insulin-dependent diabetes mellitus and in the development of thyroid hypertrophy, or cause spontaneous abortions and respiratory tract infections. Indirect health hazards can occur as a consequence of algal toxins, causing nausea, vomiting, diarrhoea, pneumonia, gastroenteritis, hepatoenteritis, muscular cramps, and several poisoning syndromes (paralytic shellfish poisoning, neurotoxic shellfish poisoning, amnesic shellfish poisoning). Other indirect health hazards can also come from the potential relationship between inorganic nitrogen pollution and human infectious diseases (malaria, cholera). Human sickness and death, extensive kills of aquatic animals, and other negative effects, can have elevated costs on human economy, with the recreation and tourism industry suffering the most important economic impacts, at least locally. It is concluded that levels of total nitrogen lower than 0.5-1.0 mg TN/L could prevent aquatic ecosystems (excluding those ecosystems with naturally high N levels) from developing acidification and eutrophication, at least by inorganic nitrogen pollution. Those relatively low TN levels could also protect aquatic animals against the toxicity of inorganic nitrogenous compounds since, in the absence of eutrophication, surface waters usually present relatively high concentrations of dissolved oxygen, most inorganic reactive nitrogen being in the form of nitrate. Additionally, human health and economy would be safer from the adverse effects of inorganic nitrogen pollution.     

Worldwide occurrence and effects of antifouling paint booster biocides in the aquatic environment: a review

Organic booster biocides were recently introduced as alternatives to organotin compounds in antifouling products, after restrictions imposed on the use of tributyltin (TBT) in 1987. Replacement products are generally based on copper metal oxides and organic biocides. This ban has led to an increase in alternative coating products containing the above biocides. The most commonly used biocides in antifouling paints are: Irgarol 1051, diuron, Sea-nine 211, dichlofluanid, chlorothalonil, zinc pyrithione, TCMS (2,3,3,6-tetrachloro-4-methylsulfonyl) pyridine, TCMTB [2-(thiocyanomethylthio) benzothiazole], and zineb. Since 1993, several studies have demonstrated the presence of these biocides in European coastal environment as a result of their increased use. More recently, the presence of these biocides was also revealed in waters from Japan, United States, Singapore, Australia and Bermuda. This paper reviews the currently available data on the occurrence of these biocides in the aquatic environment. Some data dealing with the environmental fate, partitioning, behaviour and risk assessment of antifouling paint booster biocides are also reported in order to discuss the detected levels of contamination.     

Fate and transport of pathogens in lakes and reservoirs

Outbreaks of water-borne disease via public water supplies continue to be reported in developed countries even though there is increased awareness of, and treatment for, pathogen contamination. Pathogen episodes in lakes and reservoirs are often associated with rain events, and the riverine inflow is considered to be major source of pathogens. Consequently, the behaviour of these inflows is of particular importance in determining pathogen transport and distribution. Inflows are controlled by their density relative to that of the lake, such that warm inflows will flow over the surface of the lake as a buoyant surface flow and cold, dense inflows will sink beneath the lake water where they will flow along the bathymetry towards the deepest point. The fate of pathogens is determined by loss processes including settling and inactivation by temperature, UV and grazing. The general trend is for the insertion timescale to be shortest, followed by sedimentation losses and temperature inactivity. The fate of Cryptosporidium due to UV light inactivation can occur at opposite ends of the scale, depending on the location of the oocysts in the water column and the extinction coefficient for UV light. For this reason, the extinction coefficient for UV light appears to be a vitally important parameter for determining the risk of Cryptosporidium contamination. For risk assessment of pathogens in supply reservoirs, it is important to understand the role of hydrodynamics in determining the timescale of transport to the off-take relative to the timescale of inactivation. The characteristics of the riverine intrusion must also be considered when designing a sampling program for pathogens. A risk management framework is presented that accounts for pathogen fate and transport for reservoirs.