Assessment of the occurrence and potential risks of pharmaceuticals and their metabolites in fish and water using liquid chromatography mass spectrometry

A comprehensive method for the analysis of 11 target pharmaceuticals representing multiple therapeutic classes was developed for biological tissues (fish) and water. Water samples were extracted using solid phase extraction (SPE), while fish tissue homogenates were extracted using accelerated solvent extraction (ASE) followed by mixed-mode cation exchange SPE cleanup and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Among the 11 target pharmaceuticals analyzed, trimethoprim, caffeine, sulfamethoxazole, diphenhydramine, diltiazem, carbamazepine, erythromycin and fluoxetine were consistently detected in reclaimed water. On the other hand, caffeine, diphenhydramine and carbamazepine were consistently detected in fish and surface water samples. In order to understand the uptake and depuration of pharmaceuticals as well as bioconcentration factors (BCFs) under the worst-case conditions, mosquito fish were exposed to reclaimed water under static-renewal for 7 days, followed by a 14-day depuration phase in clean water. Characterization of the exposure media revealed the presence of 26 pharmaceuticals while 5 pharmaceuticals including caffeine, diphenhydramine, diltiazem, carbamazepine, and ibuprofen were present in the organisms as early as 5 h from the start of the exposure. Liquid chromatography ultra-high resolution Orbitrap mass spectrometry was explored as a tool to identify and quantify phase II pharmaceutical metabolites in reclaimed water. The resulting data confirmed the presence of acetyl-sulfamethoxazole and sulfamethoxazole glucuronide in reclaimed water. To my knowledge, this is the first known report of sulfamethoxazole glucuronide surviving intact through wastewater treatment plants and occurring in environmental water samples. Finally, five bioaccumulative pharmaceuticals including caffeine, carbamazepine, diltiazem, diphenhydramine and ibuprofen detected in reclaimed water were investigated regarding the acute and chronic risks to aquatic organisms. The results indicated a low potential risk of carbamazepine even under the worst case exposure scenario. Given the dilution factors that affect environmental releases, the risk of exposure to carbamazepine will be even more reduced.

Assessment of the occurrence and potential risks of antibiotics and their metabolites in South Florida waters using liquid chromatography tandem mass spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. ^ The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can't be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications.^ Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. ^ Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.^

A State and Territorial Survey Regarding Utilization of Environmental Health Shelter Assessments during Disasters, and a Secondary Analysis of Available Shelter Assessment Data

Disasters are complex events characterized by damage to key infrastructure and population displacements into disaster shelters. Assessing the living environment in shelters during disasters is a crucial health security concern. Until now, jurisdictional knowledge and preparedness on those assessment methods, or deficiencies found in shelters is limited. A cross-sectional survey (STUSA survey) ascertained knowledge and preparedness for those assessments in all 50 states, DC, and 5 US territories. Descriptive analysis of overall knowledge and preparedness was performed. Fisher’s exact statistics analyzed differences between two groups: jurisdiction type and population size. Two logistic regression models analyzed earthquakes and hurricane risks as predictors of knowledge and preparedness. A convenience sample of state shelter assessments records (n=116) was analyzed to describe environmental health deficiencies found during selected events. Overall, 55 (98%) of jurisdictions responded (states and territories) and appeared to be knowledgeable of these assessments (states 92%, territories 100%, p = 1.000), and engaged in disaster planning with shelter partners (states 96%, territories 83%, p = 0.564). Few had shelter assessment procedures (states 53%, territories 50%, p = 1.000); or training in disaster shelter assessments (states 41%, 60% territories, p = 0.638). Knowledge or preparedness was not predicted by disaster risks, population size, and jurisdiction type in neither model. Knowledge: hurricane (Adjusted OR 0.69, 95% C.I. 0.06-7.88); earthquake (OR 0.82, 95% C.I. 0.17-4.06); and both risks (OR 1.44, 95% C.I. 0.24-8.63); preparedness model: hurricane (OR 1.91, 95% C.I. 0.06-20.69); earthquake (OR 0.47, 95% C.I. 0.7-3.17); and both risks (OR 0.50, 95% C.I. 0.06-3.94). Environmental health deficiencies documented in shelter assessments occurred mostly in: sanitation (30%); facility (17%); food (15%); and sleeping areas (12%); and during ice storms and tornadoes. More research is needed in the area of environmental health assessments of disaster shelters, particularly, in those areas that may provide better insight into the living environment of all shelter occupants and potential effects in disaster morbidity and mortality. Also, to evaluate the effectiveness and usefulness of these assessments methods and the data available on environmental health deficiencies in risk management to protect those at greater risk in shelter facilities during disasters.

Environmental liabilities in insolvency:an area ripe for reform?

Purpose: The paper examines the question whether legislative reform is the ‘silver bullet’ for the problems generated by the failure of a company which is exposed to claims arising from the non-fulfilment of its environmental obligations. The limited capacity of the United Kingdom insolvency regime to facilitate the fulfilment of a debtor company's environmental obligations is often illustrated with reference to some significant judicial decisions. However, no real picture has emerged of the frequency with which these issues arise, based on which firm proposals for reform could be advanced. This paper argues that greater regard should be paid to existing mechanisms which provide a means of enabling insolvency risks to be managed or minimised, as these point towards the scope for these issues to be resolved through the environmental protection framework rather than through reliance on company and/or insolvency law. Design/methodology/approach: Research was conducted into the statutory and non-statutory regulations (such as statutory guidance), and case law principles, which underpin the treatment of the claims against an insolvent (or potentially insolvent) company resulting from its environmental activities. This included research into policies which have a bearing on this area, developed through governmental and civic consultations and studies. Findings: The paper concludes that the likelihood of a case for legislative reform being made out is weak, and the focus should accordingly shift to strengthening the effectiveness of existing law, policy and practice. Originality/value: This paper is the first (in the United Kingdom context) to challenge the perceived need for reform in this area, engaging with recent examples of such corporate failures and the impact of recent legislative and policy developments.

Taking risks in the face of uncertainty:an exploratory analysis of green innovation

The relationship between uncertainty and firms’ risk-taking behaviour has been a focus of investigation since early discussion of the nature of enterprise activity. Here, we focus on how firms’ perceptions of environmental uncertainty and their perceptions of the risks involved impact on their willingness to undertake green innovation. Analysis is based on a cross-sectional survey of UK food companies undertaken in 2008. The results reinforce the relationship between perceived environmental uncertainty and perceived innovation risk and emphasise the importance of macro-uncertainty in shaping firms’ willingness to undertake green innovation. The perceived (market-related) riskiness of innovation also positively influences the probability of innovating, suggesting either a proactive approach to stimulating market disruption or an opportunistic approach to innovation leadership.

Oil spill environmental risk assessment of stationary sources in offshore zones (Innovating a mathematical model)

Nowadays, risks arising from the rapid development of oil and gas industries are significantly increasing. As a result, one of the main concerns of either industrial or environmental managers is the identification and assessment of such risks in order to develop and maintain appropriate proactive measures. Oil spill from stationary sources in offshore zones is one of the accidents resulting in several adverse impacts on marine ecosystems. Considering a site's current situation and relevant requirements and standards, risk assessment process is not only capable of recognizing the probable causes of accidents but also of estimating the probability of occurrence and the severity of consequences. In this way, results of risk assessment would help managers and decision makers create and employ proper control methods. Most of the represented models for risk assessment of oil spills are achieved on the basis of accurate data bases and analysis of historical data, but unfortunately such data bases are not accessible in most of the zones, especially in developing countries, or else they are newly established and not applicable yet. This issue reveals the necessity of using Expert Systems and Fuzzy Set Theory. By using such systems it will be possible to formulize the specialty and experience of several experts and specialists who have been working in petroliferous areas for several years. On the other hand, in developing countries often the damages to environment and environmental resources are not considered as risk assessment priorities and they are approximately under-estimated. For this reason, the proposed model in this research is specially addressing the environmental risk of oil spills from stationary sources in offshore zones.

Assessment of the Occurrence and Potential Risks of Pharmaceuticals and their Metabolites in Fish and Water Using Liquid Chromatography Mass Spectrometry

A comprehensive method for the analysis of 11 target pharmaceuticals representing multiple therapeutic classes was developed for biological tissues (fish) and water. Water samples were extracted using solid phase extraction (SPE), while fish tissue homogenates were extracted using accelerated solvent extraction (ASE) followed by mixed-mode cation exchange SPE cleanup and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Among the 11 target pharmaceuticals analyzed, trimethoprim, caffeine, sulfamethoxazole, diphenhydramine, diltiazem, carbamazepine, erythromycin and fluoxetine were consistently detected in reclaimed water. On the other hand, caffeine, diphenhydramine and carbamazepine were consistently detected in fish and surface water samples. In order to understand the uptake and depuration of pharmaceuticals as well as bioconcentration factors (BCFs) under the worst-case conditions, mosquito fish were exposed to reclaimed water under static-renewal for 7 days, followed by a 14-day depuration phase in clean water. Characterization of the exposure media revealed the presence of 26 pharmaceuticals while 5 pharmaceuticals including caffeine, diphenhydramine, diltiazem, carbamazepine, and ibuprofen were present in the organisms as early as 5 h from the start of the exposure. Liquid chromatography ultra-high resolution Orbitrap mass spectrometry was explored as a tool to identify and quantify phase II pharmaceutical metabolites in reclaimed water. The resulting data confirmed the presence of acetyl-sulfamethoxazole and sulfamethoxazole glucuronide in reclaimed water. To my knowledge, this is the first known report of sulfamethoxazole glucuronide surviving intact through wastewater treatment plants and occurring in environmental water samples. Finally, five bioaccumulative pharmaceuticals including caffeine, carbamazepine, diltiazem, diphenhydramine and ibuprofen detected in reclaimed water were investigated regarding the acute and chronic risks to aquatic organisms. The results indicated a low potential risk of carbamazepine even under the worst case exposure scenario. Given the dilution factors that affect environmental releases, the risk of exposure to carbamazepine will be even more reduced.

Assessment of the Occurrence and Potential Risks of Antibiotics and their Metabolites in South Florida Waters Using Liquid Chromatography Tandem Mass Spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can’t be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications. Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.

A Multi-Criteria GIS-Based Route Selection Tool for Hazardous Material Transport: Consideration of Environmental Consequence, Traffic Congestions and Costs

Hazardous materials are substances that, if not regulated, can pose a threat to human populations and their environmental health, safety or property when transported in commerce. About 1.5 million tons of hazardous material shipments are transported by truck in the US annually, with a steady increase of approximately 5% per year. The objective of this study was to develop a routing tool for hazardous material transport in order to facilitate reduced environmental impacts and less transportation difficulties, yet would also find paths that were still compelling for the shipping carriers as a matter of trucking cost. The study started with identification of inhalation hazard impact zones and explosion protective areas around the location of hypothetical hazardous material releases, considering different parameters (i.e., chemicals characteristics, release quantities, atmospheric condition, etc.). Results showed that depending on the quantity of release, chemical, and atmospheric stability (a function of wind speed, meteorology, sky cover, time and location of accidents, etc.) the consequence of these incidents can differ. The study was extended by selection of other evaluation criteria for further investigation because health risk as an evaluation criterion would not be the only concern in selection of routes. Transportation difficulties (i.e., road blockage and congestion) were incorporated as important factor due to their indirect impact/cost on the users of transportation networks. Trucking costs were also considered as one of the primary criteria in selection of hazardous material paths; otherwise the suggested routes would have not been convincing for the shipping companies. The last but not least criterion was proximity of public places to the routes. The approach evolved from a simple framework to a complicated and efficient GIS-based tool able to investigate transportation networks of any given study area, and capable of generating best routing options for cargos. The suggested tool uses a multi-criteria-decision-making method, which considers the priorities of the decision makers in choosing the cargo routes. Comparison of the routing options based on each criterion and also the overall suitableness of the path in regards to all the criteria (using a multi-criteria-decision-making method) showed that using similar tools as the one proposed by this study can provide decision makers insights in the area of hazardous material transport. This tool shows the probable consequences of considering each path in a very easily understandable way; in the formats of maps and tables, which makes the tradeoffs of costs and risks considerably simpler, as in some cases slightly compromising on trucking cost may drastically decrease the probable health risk and/or traffic difficulties. This will not only be rewarding to the community by making cities safer places to live, but also can be beneficial to shipping companies by allowing them to advertise as environmental friendly conveyors.

Surveillance risks in IoT applied to smart cities

Nowadays, cities deal with unprecedented pollution and overpopulation problems, and Internet of Things (IoT) technologies are supporting them in facing these issues and becoming increasingly smart. IoT sensors embedded in public infrastructure can provide granular data on the urban environment, and help public authorities to make their cities more sustainable and efficient. Nonetheless, this pervasive data collection also raises high surveillance risks, jeopardizing privacy and data protection rights. Against this backdrop, this thesis addresses how IoT surveillance technologies can be implemented in a legally compliant and ethically acceptable fashion in smart cities. An interdisciplinary approach is embraced to investigate this question, combining doctrinal legal research (on privacy, data protection, criminal procedure) with insights from philosophy, governance, and urban studies. The fundamental normative argument of this work is that surveillance constitutes a necessary feature of modern information societies. Nonetheless, as the complexity of surveillance phenomena increases, there emerges a need to develop more fine-attuned proportionality assessments to ensure a legitimate implementation of monitoring technologies. This research tackles this gap from different perspectives, analyzing the EU data protection legislation and the United States and European case law on privacy expectations and surveillance. Specifically, a coherent multi-factor test assessing privacy expectations in public IoT environments and a surveillance taxonomy are proposed to inform proportionality assessments of surveillance initiatives in smart cities. These insights are also applied to four use cases: facial recognition technologies, drones, environmental policing, and smart nudging. Lastly, the investigation examines competing data governance models in the digital domain and the smart city, reviewing the EU upcoming data governance framework. It is argued that, despite the stated policy goals, the balance of interests may often favor corporate strategies in data sharing, to the detriment of common good uses of data in the urban context.

A framework for the environmental risk assessment of offshore carbon storage in CCS systems

Carbon capture and storage (CCS) represents an interesting climate mitigation option, however, as for any other human activity, there is the impelling need to assess and manage the associated risks. This study specifically addresses the marine environmental risk posed by CO2 leakages associated to CCS subsea engineering system, meant as offshore pipelines and injection / plugged and abandoned wells. The aim of this thesis work is to start approaching the development of a complete and standardized practical procedure to perform a quantified environmental risk assessment for CCS, with reference to the specific activities mentioned above. Such an effort would be of extreme relevance not only for companies willing to implement CCS, as a methodological guidance, but also, by uniformizing the ERA procedure, to begin changing people’s perception about CCS, that happens to be often discredited due to the evident lack of comprehensive and systematic methods to assess the impacts on the marine environment. The backbone structure of the framework developed consists on the integration of ERA’s main steps and those belonging to the quantified risk assessment (QRA), in the aim of quantitatively characterizing risk and describing it as a combination of magnitude of the consequences and their frequency. The framework developed by this work is, however, at a high level, as not every single aspect has been dealt with in the required detail. Thus, several alternative options are presented to be considered for use depending on the situation. Further specific studies should address their accuracy and efficiency and solve the knowledge gaps emerged, in order to establish and validate a final and complete procedure. Regardless of the knowledge gaps and uncertainties, that surely need to be addressed, this preliminary framework already finds some relevance in on field applications, as a non-stringent guidance to perform CCS ERA, and it constitutes the foundation of the final framework.

[lean Production And Psychosocial Risks: The Case Of A Multinational Merger In A Metallurgical Company In Brazil].

This study focused on the method known as lean production as a work-related psychosocial risk factor in a Brazilian multinational auto parts company after its merger with other multinational companies. The authors conducted a qualitative analysis of two time points: the first using on-site observation and key interviews with managers and workers during implementation of lean production in 1996; the second, 16 years later, comparing data from a document search in labor inspection records from the Ministry of Labor and Employment and legal proceedings initiated by the Office of the Public Prosecutor for Labor Affairs. The merger led to layoffs, replacements, and an increase in the workday. A class action suit was filed on grounds of aggravated working conditions. The new production model led to psychosocial risks that increased the need for workers' health precautions when changes in the production process introduced new and increased risks of physical and mental illnesses.

Long-term Prospects For The Environmental Profile Of Advanced Sugar Cane Ethanol.

This work assessed the environmental impacts of the production and use of 1 MJ of hydrous ethanol (E100) in Brazil in prospective scenarios (2020-2030), considering the deployment of technologies currently under development and better agricultural practices. The life cycle assessment technique was employed using the CML method for the life cycle impact assessment and the Monte Carlo method for the uncertainty analysis. Abiotic depletion, global warming, human toxicity, ecotoxicity, photochemical oxidation, acidification, and eutrophication were the environmental impacts categories analyzed. Results indicate that the proposed improvements (especially no-til farming-scenarios s2 and s4) would lead to environmental benefits in prospective scenarios compared to the current ethanol production (scenario s0). Combined first and second generation ethanol production (scenarios s3 and s4) would require less agricultural land but would not perform better than the projected first generation ethanol, although the uncertainties are relatively high. The best use of 1 ha of sugar cane was also assessed, considering the displacement of the conventional products by ethanol and electricity. No-til practices combined with the production of first generation ethanol and electricity (scenario s2) would lead to the largest mitigation effects for global warming and abiotic depletion. For the remaining categories, emissions would not be mitigated with the utilization of the sugar cane products. However, this conclusion is sensitive to the displaced electricity sources.

Genetic Diversity Of Giardia Duodenalis: Multilocus Genotyping Reveals Zoonotic Potential Between Clinical And Environmental Sources In A Metropolitan Region Of Brazil.

Giardia duodenalis is a flagellate protozoan that parasitizes humans and several other mammals. Protozoan contamination has been regularly documented at important environmental sites, although most of these studies were performed at the species level. There is a lack of studies that correlate environmental contamination and clinical infections in the same region. The aim of this study is to evaluate the genetic diversity of a set of clinical and environmental samples and to use the obtained data to characterize the genetic profile of the distribution of G. duodenalis and the potential for zoonotic transmission in a metropolitan region of Brazil. The genetic assemblages and subtypes of G. duodenalis isolates obtained from hospitals, a veterinary clinic, a day-care center and important environmental sites were determined via multilocus sequence-based genotyping using three unlinked gene loci. Cysts of Giardia were detected at all of the environmental sites. Mixed assemblages were detected in 25% of the total samples, and an elevated number of haplotypes was identified. The main haplotypes were shared among the groups, and new subtypes were identified at all loci. Ten multilocus genotypes were identified: 7 for assemblage A and 3 for assemblage B. There is persistent G. duodenalis contamination at important environmental sites in the city. The identified mixed assemblages likely represent mixed infections, suggesting high endemicity of Giardia in these hosts. Most Giardia isolates obtained in this study displayed zoonotic potential. The high degree of genetic diversity in the isolates obtained from both clinical and environmental samples suggests that multiple sources of infection are likely responsible for the detected contamination events. The finding that many multilocus genotypes (MLGs) and haplotypes are shared by different groups suggests that these sources of infection may be related and indicates that there is a notable risk of human infection caused by Giardia in this region.

Environmental Controls In The Water Use Patterns Of A Tropical Cloud Forest Tree Species, Drimys Brasiliensis (winteraceae).

Trees from tropical montane cloud forest (TMCF) display very dynamic patterns of water use. They are capable of downwards water transport towards the soil during leaf-wetting events, likely a consequence of foliar water uptake (FWU), as well as high rates of night-time transpiration (Enight) during drier nights. These two processes might represent important sources of water losses and gains to the plant, but little is known about the environmental factors controlling these water fluxes. We evaluated how contrasting atmospheric and soil water conditions control diurnal, nocturnal and seasonal dynamics of sap flow in Drimys brasiliensis (Miers), a common Neotropical cloud forest species. We monitored the seasonal variation of soil water content, micrometeorological conditions and sap flow of D. brasiliensis trees in the field during wet and dry seasons. We also conducted a greenhouse experiment exposing D. brasiliensis saplings under contrasting soil water conditions to deuterium-labelled fog water. We found that during the night D. brasiliensis possesses heightened stomatal sensitivity to soil drought and vapour pressure deficit, which reduces night-time water loss. Leaf-wetting events had a strong suppressive effect on tree transpiration (E). Foliar water uptake increased in magnitude with drier soil and during longer leaf-wetting events. The difference between diurnal and nocturnal stomatal behaviour in D. brasiliensis could be attributed to an optimization of carbon gain when leaves are dry, as well as minimization of nocturnal water loss. The leaf-wetting events on the other hand seem important to D. brasiliensis water balance, especially during soil droughts, both by suppressing tree transpiration (E) and as a small additional water supply through FWU. Our results suggest that decreases in leaf-wetting events in TMCF might increase D. brasiliensis water loss and decrease its water gains, which could compromise its ecophysiological performance and survival during dry periods.