Inbred SJL mice recapitulate human resistance to Cryptococcus infection due to differential immune activation.

IMPORTANCE: Cryptococcosis studies often utilize the common C57BL/6J mouse model. Unfortunately, infection in these mice fails to replicate the basic course of human disease, particularly hampering immunological studies. This work demonstrates that SJL/J mice can recapitulate human infection better than other mouse strains. The immunological response to Cryptococcus infection in SJL/J mice was markedly different from C57BL/6J and much more productive in combating this infection. Characterization of infected mice demonstrated strain-specific genetic linkage and differential regulation of multiple important immune-relevant genes in response to Cryptococcus infection. While our results validate many of the previously identified immunological features of cryptococcosis, we also demonstrate limitations from previous mouse models as they may be less translatable to human disease. We concluded that SJL/J mice more faithfully recapitulate human cryptococcosis serving as an exciting new animal model for immunological and genetic studies.

Improving the design and conduct of aquatic toxicity studies with oils based on 20 years of CROSERF experience.

Laboratory toxicity testing is a key tool used in oil spill science, spill effects assessment, and mitigation strategy decisions to minimize environmental impacts. A major consideration in oil toxicity testing is how to replicate real-world spill conditions, oil types, weathering states, receptor organisms, and modifying environmental factors under laboratory conditions. Oils and petroleum-derived products are comprised of thousands of compounds with different physicochemical and toxicological properties, and this leads to challenges in conducting and interpreting oil toxicity studies. Experimental methods used to mix oils with aqueous test media have been shown to influence the aqueous-phase hydrocarbon composition and concentrations, hydrocarbon phase distribution (i.e., dissolved phase versus in oil droplets), and the stability of oil:water solutions which, in turn, influence the bioavailability and toxicity of the oil containing media. Studies have shown that differences in experimental methods can lead to divergent test results. Therefore, it is imperative to standardize the methods used to prepare oil:water solutions in order to improve the realism and comparability of laboratory tests. The CROSERF methodology, originally published in 2005, was developed as a standardized method to prepare oil:water solutions for testing and evaluating dispersants and dispersed oil. However, it was found equally applicable for use in testing oil-derived petroleum substances. The goals of the current effort were to: (1) build upon two decades of experience to update existing CROSERF guidance for conducting aquatic toxicity tests and (2) to improve the design of laboratory toxicity studies for use in hazard evaluation and development of quantitative effects models that can then be applied in spill assessment. Key experimental design considerations discussed include species selection (standard vs field collected), test substance (single compound vs whole oil), exposure regime (static vs flow-through) and duration, exposure metrics, toxicity endpoints, and quality assurance and control.

The Challenges of Characterizing the Zooplankton Community Response Following Simulated Spills of Diluted Bitumen into Boreal Lake Limnocorrals.

We attempted to characterize zooplankton community response following spills of the unconventional crude oil, diluted bitumen (dilbit), into 10-m diameter, ~ 100 m3, ~ 1.5-m deep boreal lake limnocorrals, including two controls and seven dilbit treatments ranging from 1.5 to 180 L (1:100,000 to 1:1,000 v/v, dilbit:water). Community composition and abundances were monitored weekly to bi-weekly over three months. Total zooplankton biomass and abundance seemingly collapsed in all limnocorrals, regardless of treatment, though some rotifer species persisted. As a result, it was not possible to determine the impacts of dilbit. We theorize several potential non-oil-related reasons for the sudden community collapse - including elevated zinc levels, fish grazing pressures, and sampling biases - and provide guidance for future work using in-lake enclosures.

Surface oil is the primary driver of macroinvertebrate impacts following spills of diluted bitumen in freshwater.

The response of freshwater invertebrates following accidental releases of oil is not well understood. This knowledge gap is more substantial for unconventional oils such as diluted bitumen (dilbit). We evaluated the effects of dilbit on insect emergence and benthic invertebrates by conducting experimental spills in limnocorrals (10-m diameter; ~100-m3) deployed in a boreal lake at the IISD-Experimental Lakes Area, Canada. The study included seven dilbit treatments (spill volumes ranged from 1.5 L [1:66,000, oil:water, v/v] to 180 L [1:590, oil:water, v/v]), two controls, and additional lake reference sites, monitored for 11 weeks. Invertebrate emergence declined at the community level following oil addition in a significantly volume-dependent manner, and by 93-100 % over the 11 weeks following the spill in the highest treatment. Dilbit altered community structure of benthic invertebrates, but not abundance. One-year post-spill and following oil removal using traditional skimming and absorption techniques, benthic richness and abundance were greater among all treatments than the previous year. These results indicate that recovery in community composition is possible following oil removal from a lake ecosystem. Research is needed concerning the mechanisms by which surface oil directly affect adult invertebrates, whether through limiting oviposition, limiting emergence, or both. The response of benthic communities to sediment tar mats is also warranted.

Caffeine and paraxanthine in aquatic systems: Global exposure distributions and probabilistic risk assessment.

This study presents one of the most complete applications of probabilistic methodologies to the risk assessment of emerging contaminants. Perhaps the most data-rich of these compounds, caffeine, as well as its main metabolite (paraxanthine), were selected for this study. Information for a total of 29,132 individual caffeine and 7442 paraxanthine samples was compiled, including samples where the compounds were not detected. The inclusion of non-detect samples (as censored data) in the estimation of environmental exposure distributions (EEDs) allowed for a realistic characterization of the global presence of these compounds in aquatic systems. EEDs were compared to species sensitivity distributions (SSDs), when possible, in order to calculate joint probability curves (JPCs) to describe the risk to aquatic organisms. This way, it was determined that unacceptable environmental risk (defined as 5% of the species being potentially exposed to concentrations able to cause effects in>5% of the cases) could be expected from chronic exposure to caffeine from effluent (28.4% of the cases), surface water (6.7% of the cases) and estuary water (5.4% of the cases). Probability of exceedance of acute predicted no-effect concentrations (PNECs) for paraxanthine were higher than 5% for all assessed matrices except for drinking water and ground water, however no experimental effects data was available for paraxanthine, resulting in a precautionary deterministic hazard assessment for this compound. Given the chemical similarities between both compounds, real effect thresholds, and thus risk, for paraxanthine, would be expected to be close to those observed for caffeine. Negligible Human health risk from exposure to caffeine via drinking or groundwater is expected from the compiled data.

Drugs of abuse and benzodiazepines in the Madrid Region (Central Spain): seasonal variation in river waters, occurrence in tap water and potential environmental and human risk.

This work analyzes the seasonal variation (winter and summer) of ten drugs of abuse, six metabolites and three benzodiazepines in surface waters from the Jarama and Manzanares Rivers in the Madrid Region, the most densely populated area in Spain. The occurrence of these compounds in tap water in this region is also investigated and a preliminary human health risk characterization performed for those substances found in tap water. Finally, a screening level risk assessment that combines the measured environmental concentrations (MECs) with dose-response data to estimate Hazard Quotients (HQs) for the compounds studied is also presented. The results of this study show the presence of fourteen out of the nineteen compounds analyzed in winter and twelve of them in summer. The most ubiquitous compounds, with a frequency of detection of 100% in both seasons, were the cocaine metabolite benzoylecgonine (BE), the amphetamine-type stimulant (ATS) ephedrine (EPH), the opioid methadone (METH), the METH metabolite 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), and the three benzodiazepines investigated, namely alprazolam (ALP), diazepam (DIA) and lorazepam (LOR). The highest concentrations observed corresponded to EPH (1020ngL(-1) in winter and 250ngL(-1) in summer). The only compounds not detected in both seasons were heroin (HER) and its metabolite 6-acetylmorphine (6ACM), lysergic acid diethylamide (LSD) and its metabolite 2-oxo-3-hydroxy-LSD (O-H-LSD), and Δ(9)-tetrahydrocannabinol (THC). In terms of overall concentration, all sampling points presented higher concentrations in winter than in summer. Statistical analyses performed to gather evidence concerning occasional seasonal differences in the concentrations of individual substances between summer and winter showed statistically significantly higher concentrations (p<0.05) of BE, EPH and the opioid morphine (MOR) in winter than in summer. Two out of the nineteen compounds studied, namely cocaine (CO) and EPH, were detected in tap water from one sampling point at concentrations of 1.61 and 0.29ngL(-1), respectively. The preliminary human health risk characterization showed that no toxic effects could be expected at the detected concentration level in tap water. The screening level risk assessment showed that MOR, EDDP and the THC metabolite 11-nor-9-carboxy-Δ(9)-tetrahydrocannabinol (THC-COOH) were present in at least one of the sampling sites in a concentration leading to a Hazard Quotient (HQ) value between 1.0 and 10.0, thus indicating some possible adverse effects. The cumulative HQ or Toxic units (TUs) calculated for each of the groups studied showed that opioids and cannabinoids were present at concentrations high enough to potentially generate some adverse effects on at least one sampling point.

Seasonal variation of pharmaceutically active compounds in surface (Tagus River) and tap water (Central Spain).

Numerous studies have shown the presence of pharmaceutically active compounds (PhACs) in different environmental compartments, for example, in surface water or wastewater ranging from nanograms per litre to micrograms per litre. Likewise, some recent studies have pointed to seasonal variability, thus indicating that PhAcs concentrations in the aquatic environment may depend on the time of year. This work intended to find out (1) whether Tagus fluvial and drinking water were polluted with different groups of PhACs and (2) if their concentrations differed between winter and summer seasons. From the 58 substances analysed, 41 were found belonging to the main therapeutic groups. Statistical differences were seen for antibacterials, antidepressants, anxiolytics, antiepileptics, and cardiovascular drugs, with higher concentrations being detected in winter than in summer. These results might indicate that the PhACs analysed in this study undergo lower environmental degradation in winter than in summer. In order to confirm these initial results, a continuous monitoring should be performed especially on those PhACs that either because of an elevated consumption or an intrinsic chemical persistence are poorly degraded during winter months due to low temperatures and solar irradiation. It is especially important to identify which of these specific PhACs are in order to recommend their substitution by equally effective and safe substances but also environmentally friendly.

Detection of pharmaceutically active compounds in the rivers and tap water of the Madrid Region (Spain) and potential ecotoxicological risk.

Concentrations of pharmaceutically active compounds (PhACs) in the order of ng L(-1) to µg L(-1) have been reported worldwide in waste, fluvial and even drinking water, raising concern about the efficacy of the currently employed waste water treatments in the elimination of this kind of compounds. Despite ranking 29th in terms of population, Spain is currently the 8th country on pharmaceutical prescription with an expense of 14×10(9) euros in 2008. In this context, the aim of this study was to determine the presence of 33 pharmaceutically active compounds in specific points of the main rivers of the Madrid Region (MR) as well as tap water samples from the metropolitan area of Madrid. Additionally, a screening level risk characterization by means of the Hazard Quotient (HQ) method was applied. A total of 25 pharmaceutical compounds and metabolites were detected in the 10 sampling points downstream the outlet of the major STPs of the MR. The highest concentrations were detected for the anticonvulsant carbamazepine and the stimulant caffeine. Concentrations for most of the analyzed compounds exceed levels previously reported in the literature. Moreover, we report the highest concentration of the cytostatic ifosfamide, detected for the first time in Spain in surface water. Preliminary risk characterization shows that a total of 16 compounds represent at least a low potential hazard based on their scored HQs, with five of them present in a concentration that exceeds the predicted no effect concentration (PNEC). Toxic Units calculation indicates that for all the selected sampling points high hazard is anticipated from the presence of the analyzed compounds in the measured concentrations (TUs>10). Caffeine and cotinine were detected in all (10) the analyzed tap water samples. Carbamazepine and nicotine were detected in six and venlafaxine in two samples. No studies venlafaxine in drinking water have been reported. These results clearly pinpoint the need for water quality monitoring and research in urban rivers, as well as the need for improved water treatment techniques able to eliminate this kind of compounds from the effluent waters as well as from drinking water sources.

Analysis of the presence of cardiovascular and analgesic/anti-inflammatory/antipyretic pharmaceuticals in river- and drinking-water of the Madrid Region in Spain.

Interest in the presence of pharmaceuticals in wastewater, in the water of our rivers and, to a lesser extent, in our drinking water, has been growing in recent decades. Many of these substances, currently classified as "emerging pollutants", are biologically active compounds and continuously released in effluents. As sewage treatment plants (STPs) are not adequately equipped to eliminate all of these substances completely, some are discharged directly into rivers. In Spain, as in most of its neighbouring countries, there is an elevated use of pharmaceuticals for the treatment of cardiovascular diseases (which are extremely prevalent among the older adult population) and anti-inflammatory medications, which are obtainable over the counter without a medical prescription. This study therefore sought to determine to what degree pharmaceuticals with the highest regional prescription and/or use rates, such as cardiovascular and analgesic/anti-inflammatory/antipyretic medications, were present in the principal rivers (Jarama, Manzanares, Guadarrama, Henares and Tagus) and tap-water samples of the Madrid Region (MR). Samples were taken downstream the discharge of 10 of the most important region's STPs and the most frequently used drugs in the region were analysed for. Of the 24 drugs analysed, 21 were detected at concentrations ranging from 2 ng L⁻¹ to 18 µg L⁻¹. The highest drug concentrations corresponded to ibuprofen, diclofenac, naproxen, atenolol, frusemide (furosemide), gemfibrozil and hydrochlorthiazide, and in most cases exceeded the amounts reported in the scientific literature. No traces of these groups of pharmaceuticals were detected in the drinking water analysed. On the basis of the high concentrations detected, we believe that an environmental surveillance system should be implemented to assess the continuous discharge of these pharmaceuticals and their possible ecotoxicological effects. At the same time, efforts to raise the awareness of the public about responsible use and the proper disposal of such substances at purpose-designated collection points should be increased. Furthermore sewage treatment processes should be suitably adapted to increase the rates of removal of these drugs.