Antibiotic Resistance in Marine Microbial Communities Proximal to a Florida Sewage Outfall System.

Water samples were collected at several wastewater treatment plants in southeast Florida, and water and sediment samples were collected along and around one outfall pipe, as well as along several transects extending both north and south of the respective outfall outlet. Two sets of samples were collected to address potential seasonal differences, including 38 in the wet season (June 2018) and 42 in the dry season (March 2019). Samples were screened for the presence/absence of 15 select antibiotic resistance gene targets using the polymerase chain reaction. A contrast between seasons was found, with a higher frequency of detections occurring in the wet season and fewer during the dry season. These data illustrate an anthropogenic influence on offshore microbial genetics and seasonal flux regarding associated health risks to recreational users and the regional ecosystem.

Corbicula fluminea rapidly accumulate pharmaceuticals from an effluent dependent urban stream.

Freshwater bivalve populations are stressed by watershed development at the global scale. Though pharmaceuticals released from wastewater treatment plant effluent discharges are increasingly reported to bioaccumulate in fish, an understanding of bioaccumulation in bivalves is less defined. In the present study, we examined accumulation of 12 target pharmaceuticals in C. fluminea during a 42 day in situ study in Pecan Creek, an effluent dependent wadeable stream in north central Texas, USA. Caged clams were placed at increasing distances (5 m, 643 m, 1762 m) downstream from a municipal effluent discharge and then subsampled on study days 7, 14, 28 and 42. Acetaminophen, caffeine, carbamazepine, diltiazem, diphenhydramine, fluoxetine, norfluoxetine, sertraline, desmethylsertraline, and methylphenidate were identified in C. fluminea whole body tissue homogenates via isotope dilution liquid chromatography-tandem mass spectrometry. Tissue concentrations ranged from low µg/kg (methylphenidate) to 341 µg/kg (sertraline). By study day 7, rapid and apparent pseudo-steady state accumulation of study compounds was observed in clams; this observation continued throughout the 42 d study. Notably, elevated bioaccumulation factors (L/kg) for sertraline were observed between 3361 and 6845, which highlights the importance of developing predictive bioaccumulation models for ionizable contaminants with bivalves. Future research is also necessary to understand different routes of exposure and elimination kinetics for pharmaceutical accumulation in bivalves.

Evaluation of sewage source and fate on southeast Florida coastal reefs.

Water, sponge and coral samples were collected from stations impacted by a variety of pollution sources and screened for human enteric viruses as conservative markers for human sewage. While human enteroviruses and adenoviruses were not detected, noroviruses (NoV; human genogroups I and II) were detected in 31% of samples (especially in sponge tissue). Stations near inlets were the only ones to show multiple sample types positive for NoV. Fecal indicator bacteria and enteric viruses were further evaluated at multiple inlet stations on an outgoing tide. Greatest indicator concentrations and highest prevalence of viruses were found at the mouth of the inlet and offshore in the inlet plume. Results suggest that inlets moving large volumes of water into the coastal zone with tides may be an important source of fecal contaminants. Efforts to reduce run-off or unintended release of water into the Intracoastal Waterway may lower contaminants entering sensitive coastal areas.

Severe 2010 cold-water event caused unprecedented mortality to corals of the Florida reef tract and reversed previous survivorship patterns.

BACKGROUND: Coral reefs are facing increasing pressure from natural and anthropogenic stressors that have already caused significant worldwide declines. In January 2010, coral reefs of Florida, United States, were impacted by an extreme cold-water anomaly that exposed corals to temperatures well below their reported thresholds (16°C), causing rapid coral mortality unprecedented in spatial extent and severity. METHODOLOGY/PRINCIPAL FINDINGS: Reef surveys were conducted from Martin County to the Lower Florida Keys within weeks of the anomaly. The impacts recorded were catastrophic and exceeded those of any previous disturbances in the region. Coral mortality patterns were directly correlated to in-situ and satellite-derived cold-temperature metrics. These impacts rival, in spatial extent and intensity, the impacts of the well-publicized warm-water bleaching events around the globe. The mean percent coral mortality recorded for all species and subregions was 11.5% in the 2010 winter, compared to 0.5% recorded in the previous five summers, including years like 2005 where warm-water bleaching was prevalent. Highest mean mortality (15%-39%) was documented for inshore habitats where temperatures were <11°C for prolonged periods. Increases in mortality from previous years were significant for 21 of 25 coral species, and were 1-2 orders of magnitude higher for most species. CONCLUSIONS/SIGNIFICANCE: The cold-water anomaly of January 2010 caused the worst coral mortality on record for the Florida Reef Tract, highlighting the potential catastrophic impacts that unusual but extreme climatic events can have on the persistence of coral reefs. Moreover, habitats and species most severely affected were those found in high-coral cover, inshore, shallow reef habitats previously considered the "oases" of the region, having escaped declining patterns observed for more offshore habitats. Thus, the 2010 cold-water anomaly not only caused widespread coral mortality but also reversed prior resistance and resilience patterns that will take decades to recover.

Diazinon in surface waters before and after a federally-mandated ban.

Samples collected from rural and urban streams in the City of Denton, Texas, USA were analyzed for the organophosphorus pesticide diazinon during the years preceding and following a United States Environmental Protection Agency ban on many diazinon uses. A network of 70 monitoring stations, based mainly on topography and hydrological considerations, were established within the three main watersheds of Denton. Monitoring stations were sampled monthly from March through August during periods of normal flow (baseflow), resulting in a total of 1243 samples collected during the years of 2001-2004. Pesticide concentrations were determined using commercially available enzyme-linked immunosorbent assays (ELISAs) specific for diazinon. Results from this temporally and spatially dense monitoring effort illustrated the impacts of a decrease in diazinon production during 2002, followed by a ban on most outdoor, non-agricultural diazinon retail sales imposed during 2003. The total number of samples exhibiting diazinon concentrations above the lower limits of detection (LLD) significantly decreased between 2001 through 2004 (Mantel-Haenszel Chi-Square test, p<0.0001, n=1243) and decreased significantly during the four monitoring years (Cochran-Armitage Trend test, z=-17.94, p<0.0001, n=1243). The total number of stations exhibiting at least one sample above the LLD during the four monitoring years showed similar patterns (Mantel-Haenszel Chi-Square test, p<0.0001; Cochran-Armitage Trend test, z=-3.21, p=0.0007; n=276). Results indicate that the phased reduction of outdoor, non-agricultural diazinon uses led to a highly significant decrease in surface water occurrences of this pesticide.

Chlorpyrifos in surface waters before and after a federally mandated ban.

Samples collected from rural and urban streams in the City of Denton, Texas, USA were analyzed for the organophosphorus pesticides diazinon and chlorpyrifos during the years preceding and following a United States Environmental Protection Agency ban on many chlorpyrifos uses. A network of 70 monitoring stations, based mainly on topography and hydrological considerations, were established within the three main watersheds of Denton. Monitoring stations were sampled monthly from March through August during periods of normal flow (baseflow), resulting in a total of 308 samples and 311 samples collected during 2001 and 2002, respectively. Pesticide concentrations were determined using commercially available enzyme-linked immunosorbent assays (ELISA) specific for each pesticide. Results from this temporally and spatially dense monitoring effort were used to illustrate the impacts of a ban on most chlorpyrifos sales that was imposed in December 2001. The total number of samples exhibiting concentrations above method lower limits of detection (LLD) decreased between 2001 and 2002 for both chlorpyrifos and diazinon. The total number of sites exhibiting at least one sample with concentrations above the LLD for chlorpyrifos, however, significantly decreased (Fisher's exact test, p<<0.0001, n=70) when comparing 2001 (before the ban) with 2002 (after the ban). Similar analyses indicated no significant difference (Fisher's exact test, p=0.50, n=70) in the number of sites exhibiting detectable diazinon concentration between 2001 and 2002. Our results indicate that the cessation of retail chlorpyrifos sales at the end of 2001 resulted in a highly significant decrease in the surface water occurrences of this pesticide during 2002.

Determination of select antidepressants in fish from an effluent-dominated stream.

Increasing evidence indicates widespread occurrence of pharmaceuticals and personal care products (PPCPs) in municipal effluent discharges and surface waters. Studies that characterize the fate and effects of PPCPs in aquatic systems are limited, and to our knowledge, data regarding pharmaceutical accumulation in fish of effluent-dominated ecosystems have not been previously reported. In the present study, fish populations were sampled from a reference stream and an effluent-dominated stream in north Texas, USA. Lepomis macrochirus, Ictalurus punctatus, Cyprinus carpio, and Pomoxis nigromaculatus were killed; the liver, brain, and lateral filet tissues dissected; and the tissues stored at -80 degrees C until analysis. Fish tissues were extracted using solid-phase extraction and then analyzed by gas chromatography-mass spectrometry in the negative chemical ionization mode. The selective serotonin reuptake inhibitors (SSRIs) fluoxetine and sertraline and the SSRI metabolites norfluoxetine and desmethylsertraline were detected at levels greater than 0.1 ng/g in all tissues examined from fish residing in a municipal effluent-dominated stream. To our knowledge, the present study is the first report of SSRI residues in fish residing within municipal effluent-dominated systems.

Assessing the accuracy of acoustic seabed classification for mapping coral reef environments in South Florida (Broward County, USA).

The Atlantic coast of Broward County, Florida (USA) is paralleled by a series of progressively deeper, shore-parallel coral reef communities. Two of these reef systems are drowned early Holocene coral reefs of 5 ky and 7 ky uncorrected radiocarbon age. Despite the case of access to these reefs, and their major contribution to the local economy, accurate benthic habitat maps of the area are not available. Ecological studies have shown that different benthic communities (i.e. communities composed of different biological taxa) exist along several spatial gradients on all reefs. Since these studies are limited by time and spatial extent, acoustic surveys with the QTCView V bottom classification system based on a 50 kHz transducer were used as an alternative method of producing habitat maps. From the acoustic data of a 3.1 km(2) survey area, spatial prediction maps were created for the area. These were compared with habitat maps interpreted from in situ data and Laser Airborne Depth Sounder (LADS) bathymetry, in order to ground-truth the remotely sensed data. An error matrix was used to quantitatively determine the accuracy of the acoustically derived spatial prediction model against the maps derived from the in situ and LADS data sets. Confusion analysis of 100 random points showed that the system was able to distinguish areas of reef from areas of rubble and sand with an overall accuracy of 61%. When asked to detect more subtle spatial differences, for example, those between distinct reef communities, the classification was only about 40% accurate. We discuss to what degree a synthesis of acoustic and in situ techniques can provide accurate habitat maps in coral reef environments, and conclude that acoustic methods were able to reflect the spatial extent and composition of at least three different biological communities.

Increased toxicity to Ceriodaphnia dubia in mixtures of atrazine and diazinon at environmentally realistic concentrations.

The acute toxicity of diazinon in combination with atrazine concentrations of 5, 10, 20, and 40 microg/L was evaluated using Ceriodaphnia dubia. Atrazine concentrations as low as 5 microg/L in combination with diazinon significantly increased toxicity to C. dubia compared to diazinon alone. Atrazine and diazinon residues within water samples collected from 65 subbasins throughout Denton, Texas, USA were used to assess the environmental relevance of pesticide concentrations. A geographical information system was used to examine the relationship between subbasin land uses and pesticide concentrations. Significant correlations were observed between in situ atrazine and diazinon concentrations and some subbasin land uses. Atrazine was significantly (P < 0.05) correlated to diazinon during some months. Of the 276 samples collected, 39% exceeded our experimentally derived diazinon LC(50) value, and 39% exceeded our minimum atrazine concentration of 5.0 microg/L. Results indicate the potential for increased toxicity from mixtures of compounds at environmentally realistic concentrations.

Joint acute toxicity of diazinon and copper to Ceriodaphnia dubia.

Diazinon and copper are two contaminants that are widely found in urban streams and in municipal wastewater effluents. Because these contaminants may be found concurrently, the potential for their joint toxicity is of interest, particularly with regard to toxicity testing of wastewater effluents and the ecological implications of simultaneous exposures in urban streams. Although interactions between metals are well studied, relatively little is known about interactions between metals and organophosphate compounds such as diazinon. In this study, the interaction between copper and diazinon was evaluated using cladoceran (Ceriodaphnia dubia) in 48-h static tests within laboratory water. Using toxic units derived from concurrently established 48-h median lethal concentration values (LC50) of test organisms of each individual toxicant, the effects of the mixture of copper and diazinon on the survival of C. dubia were shown to be generally nonconcentration additive (LC50 significantly greater than one toxic unit). However, evaluation of the dose-response relationship across the entire range of effect levels revealed that the mortality induced by the mixture of copper and diazinon supported concentration additivity at higher effect levels.