Patterns of metal bioaccumulation in two filter-feeding macroinvertebrates: Exposure distribution, inter-species differences and variability across developmental stages

This study focused on the metal bioaccumulation of two aquatic insects (Ephoron virgo and Hydropsyche spp.) in order to evaluate the spatial distribution of metals, the interspecific differences between both filter-feeders and the bioaccumulation dynamics during E. virgo development stages. Hg, Cd, Ni, Cr, As, Pb, Cu, Ti, Zn and Mn were quantified in insects and in suspended particulate matter (SPM) sampled downstream and upstream of a chemical plant, where more than 300,000 t of polluted sediments are deposited. Hg concentrations were one order of magnitude higher downstream of the sediment dump, which showed that the Hg pollution originated in the chemical plant. Cd, Ni, Cr, Pb, Ti, Zn and Mn in invertebrates revealed that metal pollution was present upstream in other parts of the river. Interspecific differences were observed for all metals but Mn; significantly higher concentrations were observed in E. virgo over Hydropsyche exocellata, except for Cd, which showed 10-fold higher values. Hg and Cd increased until E. virgo nymphs reached 11 mm and decreased afterwards in late instars when nymphs were about to emerge. Cr, Pb, Ti and Mn decreased along early instars followed by a steady state in late instars. Similar values were obtained for Cu, As and Zn along all instars. Sexual differences between males and females of E. virgo were observed for Cd, Cu and Mn. Hg and Cd persistence was strong across developmental stages since high concentrations were found in eggs and emerging adults. Because the behavior of different metals varied for the two species and during the developmental stages of E. virgo, care should be taken in the interpretation of insect metal concentrations when analyzing the food chain transfer of metals in river ecosystems.     

Effects of pollution on land snail abundance, size and diversity as resources for pied flycatcher, Ficedula hypoleuca

Passerine birds need extra calcium during their breeding for developing egg shells and proper growth of nestling skeleton. Land snails are an important calcium source for many passerines and human-induced changes in snail populations may pose a severe problem for breeding birds. We studied from the bird's viewpoint how air pollution affects the shell mass, abundance and diversity of land snail communities along a pollution gradient of a copper smelter. We sampled remnant snail shells from the nests of an insectivorous passerine, the pied flycatcher, Ficedula hypoleuca, to find out how the availability of land snails varies along the pollution gradient. The total snail shell mass increased towards the pollution source but declined abruptly in the vicinity of the smelter. This spatial variation in shell mass was evident also within a single snail species and could not be wholly explained by spatially varying snail numbers or species composition. Instead, the total shell mass was related to their shell size, individuals being largest at the moderately polluted areas. Smaller shell size suggests inferior growth of snails in the most heavily polluted area. Our study shows that pollution affects the diversity, abundance (available shell mass) and individual quality of land snails, posing reproductive problems for birds that rely on snails as calcium sources during breeding. There are probably both direct pollution-related (heavy metal and calcium levels) and indirect (habitat change) effects behind the observed changes in snail populations.     

Dynamics of metal availability and toxicity in historically polluted floodplain sediments

Many floodplains contain high concentrations of sediment associated contaminants that might be subjected to large changes in terms of mobility, transformation and bioavailability. Therefore, this study describes 1) changes in the redox conditions and the mobility of metals in artificially uncovered polluted floodplain sediments, 2) metal uptake by organisms and 3) colonization, succession and functioning of benthic algae on these sediments. Flooding caused long term changes in redox potential (Eh) profiles. In top layers strong gradients in redox potential established quickly, while in deeper layers changes occurred more gradually. The availability of copper as measured by Diffusive Gradients in Thinfilms (DGT) showed a consistent relationship with fluctuations in Eh. However, this relationship was restricted to deeper layers in the sediment. Within 1 week high amounts of total copper were immobilized. Differences in total copper concentrations between polluted and clean sediments became only partially apparent when comparing the available copper fraction (DGT-Cu). Introduced Tubifex shows only marginal differences in levels of accumulated Cu. Colonization, growth and succession of algal communities on polluted sediments was not impaired, most likely due to low bioavailability. It is concluded that changing environmental conditions, such as flooding, can result in stable chemical conditions with low a availability of metals and hence in a diminution of actual ecological risks.     

Ring test for whole-sediment toxicity assay with -a- benthic marine diatom

This work presents the results of an interlaboratory proficiency exercise for whole-sediment toxicity assays with the benthic marine diatom Cylindrotheca closterium. An assay protocol was established and followed by all participating laboratories. Cell growth after 72 h exposure was the endpoint used. Four sediment samples of unknown toxicity were assayed. The main problem encountered during this exercise was the differences in the cell growth of algae exposed to reference sediment. Those differences may be associated with changes in the physiological status of the initial culture due to temperature changes during transport to the other laboratories. In general, the method proposed presented good replicability (precision between replicates) and reproducibility (interlaboratory precision). Around 80% (17 out of 21) of results obtained were classified as satisfactory (Z-scores < 2). The whole-sediment assay with C. closterium presented here can be considered sufficiently successful for possible use as a standard toxicity test. The assay is simple to perform, the proposed species is ecologically relevant as an integral component of microphytobenthos, and is widely distributed around the world. These positive factors suggest that the whole-sediment assay with the benthic marine diatom C. closterium can be used as a reliable tool in marine sediment quality assessment.     

Short- and long-term dynamics of the toxic potential and genotypic structure in benthic populations of Microcystis

Microcystis colonies are known to overwinter on the surface of the sediment of freshwater ecosystems. However, little is known about the genotypic and toxicological dynamics of Microcystis populations during this benthic life stage. In this study, we report a two-year-long survey of benthic populations of Microcystis, which had spent from a few days to more than six years in the sediment. In order to avoid any interaction with the planktonic proliferations, we chose two deeply buried benthic populations, which could be easily dated. Quantitative PCR on mcyB gene and protein phosphatase inhibition assays were performed to measure their toxic potential, and their genotypic structure was assessed by Capillary Electrophoresis-Single Strand Conformation Polymorphism (CE-SSCP), based on 16S-23S Intergenic Transcribed Spacer (ITS). The microcystin content of the cells seemed to change sharply during the first few months of benthic survival, whereas this content was low and decreased steadily after several years of benthic life. No genetic selection was observed in either the proportion of potentially toxic clones or the ITS sequences for any of the populations considered. From these results, the benthic life stage of Microcystis appears to preserve the structure and the composition of the population over a far larger time scale than classical overwintering period. Finally, some genotypes were common in both of the benthic populations, even though they originated from planktonic blooms that had developed five years apart, suggesting a major overlap of planktonic proliferations in successive years.     

Measuring and modeling reduction of DDT availability to the water column and mussels following activated carbon amendment of contaminated sediment

A 28-day accumulation study demonstrated the use of mussel uptake, passive samplers, and biodynamic modeling to measure the reduction of dichlorodiphenyltrichloroethane (DDT) availability in the water column after the addition of activated carbon to contaminated sediment. Sediment collected from Lauritzen Channel, Richmond, California (16.5 mg total DDT/kg) was mixed with either virgin activated carbon or a reactivated carbon for one month, after which a 28-day laboratory exposure study was completed. Mussels (Mytilus edulis) suspended above activated carbon-treated sediment accumulated significantly less total DDT in soft tissue, 91% and 84% for virgin and reactivated carbon, respectively, as compared to untreated sediment. Mussel tissue concentrations correlated to concentrations in semipermeable membrane devices (SPMDs) and polyethylene devices (PEDs) suspended over the same sediments. A biodynamic model that incorporated DDT water concentrations, either analytically measured or estimated from PED uptake, described mussel accumulation over time. Thus, passive samplers in combination with biodynamic modeling may provide an important screening tool for assessment of filter-feeding uptake and ecological risk to water-dwelling organisms exposed to aqueous phase hydrophobic organic contaminants.     

Survey of pathogenic free-living amoebae and Legionella spp. in mud spring recreation area

Acanthamoeba, Hartmannella, and Naegleria are free-living amoebae, ubiquitous in aquatic environments. Several species within these genera are recognized as potential human pathogens. These free-living amoebae may facilitate the proliferation of their parasitical bacteria, such as Legionella. In this study, we identified Acanthamoeba, Hartmannella, Naegleria, and Legionella using various analytical procedures and investigated their occurrence at a mud spring recreation area in Taiwan. We investigated factors potentially associated with the prevalence of the pathogens, including various water types, and physical and microbiological water quality parameters. Spring water was collected from 34 sites and Acanthamoeba, Hartmannella, Naegleria, and Legionella were detected in 8.8%, 35.3%, 14.7%, and 47.1%, respectively. The identified species of Acanthamoeba included Acanthamoeba castellanii and Acanthamoeba polyphaga. Nearly all the Hartmannella isolates are identified as Hartmannella vermiformis. The Naegleria species included Naegleria australiensis and its sister groups, and two other isolates referred to a new clade of Naegleria genotypes. The Legionella species identified included unnamed Legionella genotypes, Legionella pneumophila serotype 6, uncultured Legionella spp., Legionella lytica, Legionella drancourtii, and Legionella waltersii. Significant differences (Mann-Whitney U test, P < 0.05) were observed between the presence/absence of Hartmannella and total coliforms, between the presence/absence of Naegleria and heterotrophic plate counts, and between the presence/absence of Legionella and heterotrophic plate counts. This survey confirms that pathogenic free-living amoebae and Legionella are prevalent in this Taiwanese mud spring recreation area. The presence of pathogens should be considered a potential health threat when associated with human activities in spring water.     

Survival of manure-borne E. coli in streambed sediment: Effects of temperature and sediment properties

Escherichia coli bacteria are commonly used as indicator organisms to designate of impaired surface waters and to guide the design of management practices to prevent fecal contamination of water. Stream sediments are known to serve as a reservoir and potential source of fecal bacteria (E. coli) for stream water. In agricultural watersheds, substantial numbers of E. coli may reach surface waters, and subsequently be deposited into sediments, along with fecal material in runoff from land-applied manures, grazing lands, or wildlife excreta. The objectives of this work were (a) to test the hypothesis that E. coli survival in streambed sediment in the presence of manure material will be affected by sediment texture and organic carbon content and (b) to evaluate applicability of the exponential die-off equation to the E. coli survival data in the presence of manure material. Experiments were conducted at three temperatures (4 °C, 14 °C, and 24 °C) in flow-through chambers using sediment from three locations at the Beaverdam Creek Tributary in Beltsville, Maryland mixed with dairy manure slurry in the proportion of 1000:1. Indigenous E. coli populations in sediments ranged from ca. 10¹ to 10³ MPN g⁻¹ while approx 10³ manure-borne E. coli MPN g⁻¹ were added. E. coli survived in sediments much longer than in the overlaying water. The exponential inactivation model gave an excellent approximation of data after 6-16 days from the beginning of the experiment. Slower inactivation was observed with the increase in organic carbon content in sediments with identical granulometric composition. The increase in the content of fine particles and organic carbon in sediments led not only to the slower inactivation but also to lower sensitivity of the inactivation to temperature. Streambed sediment properties have to be documented to better evaluate the role of sediments as reservoirs of E. coli that can affect microbiological stream water quality during high flow events.     

Accelerated biodegradation of pyrene and benzo[a]pyrene in the Phragmites australis rhizosphere by bacteria-root exudate interactions

We investigated the biodegradation of pyrene and benzo[a]pyrene in Phragmites australis rhizosphere sediment. We collected P. australis plants, rhizosphere sediments, and unvegetated sediments from natural aquatic sites and conducted degradation experiments using sediments spiked with pyrene or benzo[a]pyrene. Accelerated removal of pyrene and benzo[a]pyrene was observed in P. australis rhizosphere sediments with plants, whereas both compounds persisted in unvegetated sediments without plants and in autoclaved rhizosphere sediments with sterilized plants, suggesting that the accelerated removal resulted largely from biodegradation by rhizosphere bacteria. Initial densities of pyrene-utilizing bacteria were substantially higher in the rhizosphere than in unvegetated sediments, but benzo[a]pyrene-utilizing bacteria were not detected in rhizosphere sediments. Mycobacterium gilvum strains isolated from rhizosphere sediments utilized pyrene aerobically as a sole carbon source and were able to degrade benzo[a]pyrene when induced with pyrene. Phragmites australis root exudates containing phenolic compounds supported growth as a carbon source for the one Mycobacterium strain tested, and induced benzo[a]pyrene-degrading activity of the strain. The stimulatory effect on benzo[a]pyrene biodegradation and the amounts of phenolic compounds in root exudates increased when P. australis was exposed to pyrene. Our results show that Mycobacterium-root exudate interactions can accelerate biodegradation of pyrene and benzo[a]pyrene in P. australis rhizosphere sediments.     

Avian ecological risk potential in an urbanized estuary: Lower Hackensack River, New Jersey, U.S.A.

As part of a comprehensive ecological risk assessment on a broad range of species, the potential for adverse effects in birds was evaluated at a chromate ore processing residue disposal site, Study Area 7, located at the confluence of the Lower Hackensack River, Passaic River, and Upper Newark Bay. Although detection of elevated concentrations of total chromium in sediment prompted the study, it was also necessary to consider potential risks related to other chemicals present in elevated concentrations due to widespread anthropogenic activities in Upper Newark Bay and its watershed. U.S. Army Corps of Engineers' TrophicTrace^® software was used to predict exposure to avian receptors using measured concentrations of chemicals in sediment, measured concentrations of chemicals in benthic invertebrates, and measured site physicochemical parameters. The TrophicTrace^® model was improved as part of the Study Area 7 ecological risk assessment to account for (1) incidental ingestion of sediment by dabbling and diving birds, (2) area use factors for spatial overlap of wide-ranging fish species and piscivorous birds, (3) spatially-explicit utilization of the site by birds with a variety of foraging strategies, and (4) temporal patterns of site utilization by migratory species. The ecological risk assessment demonstrated that chromium in sediment does not pose unacceptable hazards to avian receptors. Potentially unacceptable hazards were indicated for several organic chemicals (i.e., pesticides, polychlorinated biphenyls, and dioxins/furans), with hazard quotients highest for Upper Newark Bay reference conditions, reflecting potential widespread chemical impacts to the estuary. The modifications to TrophicTrace^® conducted for this assessment may be prudent and applicable for improving the accuracy and realism of other assessments involving avian receptors exposed to chemicals via contaminated sediment and transfer through the food web.     

Impact of urbanization and agriculture on the occurrence of bacterial pathogens and stx genes in coastal waterbodies of central California

Fecal pollution enters coastal waters through multiple routes, many of which originate from land-based activities. Runoff from pervious and impervious land surfaces transports pollutants from land to sea and can cause impairment of coastal ocean waters. To understand how land use practices and water characteristics influence concentrations of fecal indicator bacteria (FIB) and pathogens in natural waters, fourteen coastal streams, rivers, and tidal lagoons, surrounded by variable land use and animal densities, were sampled every six weeks over two years (2008 & 2009). Fecal indicator bacteria (FIB; Escherichia coli and Enterococci) and Salmonella concentrations, the occurrence of Bacteroidales human, ruminant, and pig-specific fecal markers, E. coli O157:H7, and Shiga toxin (stx) genes present in E. coli, were measured. In addition, environmental and climatic variables (e.g., temperature, salinity, rainfall), as well as human and livestock population densities and land cover were quantified. Concentrations of FIB and Salmonella were correlated with each other, but the occurrence of host-specific Bacteroidales markers did not correlate with FIB or pathogens. FIB and Salmonella concentrations, as well as the occurrence of E. coli harboring stx genes, were positively associated with the fraction of the surrounding subwatershed that was urban, while the occurrence of E. coli O157:H7 was positively associated with the agricultural fraction. FIB and Salmonella concentrations were negatively correlated to salinity and temperature, and positively correlated to rainfall. Areal loading rates of FIB, Salmonella and E. coli O157:H7 to the coastal ocean were calculated for stream and river sites and varied with land cover, salinity, temperature, and rainfall. Results suggest that FIB and pathogen concentrations are influenced, in part, by their flux from the land, which is exacerbated during rainfall; once waterborne, bacterial persistence is affected by water temperature and salinity.     

Development of a rapid DNA extraction method and one-step nested PCR for the detection of Naegleria fowleri from the environment

Naegleria fowleri is a small free-living amoebo-flagellate found in natural and manmade thermal aquatic habitats worldwide. The organism is pathogenic to man causing fatal primary amoebic meningoencephalitis (PAM). Infection typically results from bathing in contaminated water and is usually fatal. It is, therefore, important to identify sites containing N. fowleri in the interests of preventive public health microbiology. Culture of environmental material is the conventional method for the isolation of N. fowleri but requires several days incubation and subsequent biochemical or molecular tests to confirm identification. Here, a nested one-step PCR test, in conjunction with a direct DNA extraction from water or sediment material, was developed for the rapid and reliable detection of N. fowleri from the environment. Here, the assay detected N, fowleri in 18/109 river water samples associated with a nuclear power plant in South West France and 0/10 from a similar site in the UK. Although culture of samples yielded numerous thermophilic free-living amoebae, none were N. fowleri or other thermophilic Naegleria spp. The availability of a rapid, reliable and sensitive one-step nested PCR method for the direct detection of N. fowleri from the environment may aid ecological studies and enable intervention to prevent PAM cases.     

Multi-biochemical responses of benthic macroinvertebrate species as a complementary tool to diagnose the cause of community impairment in polluted rivers

Biological indexes, based on benthic macroinvertebrate taxa, are currently used worldwide to measure river ecological quality. These indexes assign a global ecological status of the biotic community, but not necessarily may detect specific effects of water pollutants. Conversely a large set of biochemical markers measured in macroinvertebrate benthic species can detect sublethal effects and inform us about additional environmental factors that are impairing benthic communities. This is especially interesting in moderately polluted sites, where other stressors are already affecting communities but not too strongly to be detected by biotic indexes. Up to ten different markers belonging to distinct metabolic paths and 42 contaminants measured in sample collections of the caddis fly Hydropsyche exocellata were assessed across a polluted gradient in the industrialized Mediterranean River basins of Besós and Llobregat (NE, Spain). Twenty four sample collections were selected to include macroinvertebrate communities representing the five impairment degrees defined by the Spanish Environmental authorities using the biotic metrics. Results evidenced a clear deterioration of the ecological water quality parameters and benthic communities towards downstream reaches. Biochemical responses varied significantly across the studied samples and were able to differentiate samples within communities having a good and deteriorated ecological stage. Principal Component Analyses indicated that salinity was one of the major stresses affecting macroinvertebrate assemblages, whereas antioxidant and metabolizing enzymes responded differently and were closely related to high and presumably toxic levels of accumulated organic pollutants. Therefore these results indicate that the use of multiple -markers sensitive to water pollution may provide complementary information to diagnose environmental factors that are impairing macroinvertebrate communities.     

Imposex and butyltin body burden in Nassarius nitidus (Jeffreys, 1867), in coastal waters within the Basque Country (northern Spain)

Levels of imposex (superimposition of male characters, upon females) and the presence of sterile females are assessed in the gastropod Nassarius nitidus (Jeffreys, 1867), at 22 locations in the Basque Country (northern Spain). At 18 of these localities, butyltin bioaccumulation (tributyltin (TBT); dibutyltin; monobutyltin) was analysed using isotope dilution and GC-ICP-MS. Higher imposex levels and TBT body burden were found in confined harbours, with a large vessel traffic or the presence of a fishing fleet or a shipyard. For the first time, four apparently sequential types of aborted capsules are described in this species. Another novelty is the interspecific comparison between imposex intensities in sympatrically living populations of N. nitidus and N. reticulatus. This showed that sensitivity to TBT pollution of both species is relatively similar (in terms of Relative Penis Length Index). Since N. nitidus has a restricted habitat distribution due to its low presence in wave-exposed habitats, its complementary use with other species is recommended for its use in TBT monitoring programmes.     

The effects of Aphis fabae infestation on the antioxidant response and heavy metal content in field grown Philadelphus coronarius plants

The purpose of this study was to explore a possible relationship between the soil availability of metals and their concentrations in various parts of Philadelphuscoronarius plants. Moreover, the possible impact of an aphid infestation on the contamination and antioxidant response of plants from the urban environment of Kraków and the reference rural area of Zagaje Stradowskie (southern Poland) was analyzed. The contents of the glutathione, proline, non-protein − SH groups, antioxidants, and phosphorous and the levels of guaiacol peroxidase and catalase activity in leaves and shoots either infested or not by the aphid Aphis fabae Scop., were measured. The potential bioavailability of metals (Cd; Cu; Ni; Pb; Zn) in the soil and their concentrations in P. coronarius plants originating from both sites were compared.The antioxidant responses were generally elevated in the plants in the polluted area. Such reactions were additionally changed by aphid infestation. Generally, the concentrations of metals in the HNO₃ and CaCl₂ extractants of the soils from two layers at the 0-20 and 20-40 cm depths from the polluted area were higher than in those from the reference area. Such differences were found for nickel and lead (in all examined extractants), zinc (in soil extractants from the layer at 20-40 cm) and cadmium (in HNO₃ extractants). Significant positive relationships between the lead concentrations in the soil and in the plants were found. In the parts of plants from the polluted area, higher concentrations of Pb and Zn (leaves and shoots) and Cd (shoots) were recorded. The shoots and leaves of plants infested with aphids had higher concentrations of Zn but lower Pb. Moreover, their leaves had higher contaminations of Cu and Ni. In conclusion, aphids affected not only the antioxidant response of the plants but also their contamination with metals, especially contamination of the leaves.     

Infection risk assessment of diarrhea-related pathogens in a tropical canal network

A quantitative microbial risk assessment (QMRA) of Cryptosporidium, Giardia and diarrhegenic Escherichia coli (DEC) infection was performed using Monte Carlo simulations to estimate the human health risks associated with the use of canal water for recreational purposes, unrestricted and restricted irrigation in a tropical peri-urban area. Three canals receiving municipal, agricultural, and, predominantly, industrial wastewater were investigated. Identification of pathogenic protozoans revealed the major presence of Cryptosporidium hominis and both assemblages A and B of Giardia lamblia. The highest individual infection risk estimate was found to be for Giardia in an exposure scenario involving the accidental ingestion of water when swimming during the rainy season, particularly in the most polluted section, downstream of a large wholesale market. The estimated annual risks of diarrheal disease due to infection by the protozoan parasites were up to 120-fold greater than the reported disease incidence in the vicinity of the studied district and the entire Thailand, suggesting a significant host resistance to disease beyond our model's assumptions. In contrast, annual disease risk estimates for DEC were in agreement with actual cases of diarrhea in the study area.     

Haematological status of wintering great tits (Parus major) along a metal pollution gradient

In the long-term biomonitoring of wild populations inhabiting polluted areas, the use of non-destructive biomarkers as markers of condition is very important. We examined the possible effects of metal pollution on the haematological status of adult great tits (Parus major) along a well-established pollution gradient near a non-ferrous smelter in Belgium. We measured blood and feather metal concentrations and assessed the haematological status (amount of red blood cells, haemoglobin concentration, haematocrit, mean corpuscular volume and mean corpuscular haemoglobin) of adult great tits during winter at four study sites. Metal concentrations in blood and feathers indicated that cadmium and lead were the most important metals in the pollution gradient under study. Measurements of haematological parameters revealed that haemoglobin concentration, haematocrit, mean corpuscular volume and mean corpuscular haemoglobin were lower in great tits from the more polluted sites. These parameters were significantly negatively correlated with blood lead concentration. The amount of red blood cells, however, did not significantly differ among study sites. Our results indicate that the haematological status of great tits is negatively affected by metal pollution and may therefore be used as a successful biomarker for monitoring the negative impact of metal exposure in the wild.     

Introduction of the land snail Eobania vermiculata as a bioindicator organism of terrestrial pollution using a battery of biomarkers

The present study aimed to enrich the group of sentinel organisms of terrestrial pollution biomonitoring, by investigating the efficacy of the land snail Eobania vermiculata. For this reason, a package of biomarkers was performed on land snails E. vermiculata collected from polluted areas in the field or treated with heavy metals in the laboratory. The biomarkers used were neutral red lysosomal retention assay of the haemocytes, acetylcholinesterase activity in the digestive gland and the haemolymph, and metallothionein content of the digestive gland. Moreover, the morphometric changes in the lysosomal system and the morphometric alterations of the neutral lipids were also investigated. In addition, the content of cadmium, lead and copper was evaluated in the digestive gland of the snails. The results revealed appreciable alterations in the biomarker values both in field- and laboratory-conditions, accompanied by significant correlations among the biomarkers. Therefore, this exploratory study suggests the utility of E. vermiculata as a sentinel organism for biomonitoring the biologic impact of terrestrial pollution, and supports the package's efficacy of the selected biomarkers.     

Rapid decay of host-specific fecal Bacteroidales cells in seawater as measured by quantitative PCR with propidium monoazide

We investigated the persistence of feces-derived Bacteroidales cells and their DNA in seawater under natural conditions using an optimized chemical method based on co-extraction of nucleic acids with propidium monoazide (PMA), which interferes with PCR amplification of molecular markers from extracellular DNA and dead bacterial cells. The previously validated Bacteroidales assays BacUni-UCD, BacHum-UCD, BacCow-UCD, and BacCan-UCD were utilized to determine concentrations of Bacteroidales genetic markers targeting all warm-blooded animals, humans, cows and dogs, specifically, over a period of 24 d. Microcosms containing mixed feces in dialysis tubing were exposed to seawater under flow-through conditions at ambient temperature in the presence and absence of sunlight. Using a two-stage plus linear decay model, the average T₉₉ (two-log reduction) of host-specific Bacteroidales cells was 28 h, whereas that of host-specific Bacteroidales DNA was 177 h. Natural sunlight did not affect the survival of uncultivable Bacteroidales cells and their DNA with the exception of the BacCow-UCD marker. Bacteroidales DNA, as measured by quantitative PCR (qPCR) without PMA, persisted for as long as 24 d at concentrations close to the limit of detection. Culturable Enterococcus cells were detected for only 70 h, whereas Enterococcus cells measured by qPCR with and without PMA persisted for 450 h. In conclusion, measuring Bacteroidales DNA without differentiating between intact and dead cells or extracellular DNA may misinform about the extent of recent fecal pollution events, particularly in the case of multiple sources of contamination with variable temporal and spatial scales due to the relatively long persistence of DNA in the environment. In contrast, applying qPCR with and without PMA can provide data on the fate and transport of fecal Bacteroidales in water, and help implement management practices to protect recreational water quality.