Ecotoxicity of plant extracts and essential oils: A review.

Plant-based products such as essential oils and other extracts have been used for centuries due to their beneficial properties. Currently, their use is widely disseminated through a variety of industries and new applications are continuously emerging. For these reasons, they are produced industrially in large quantities and consequently they have the potential to reach the environment. However, the potential effects that these products have on the ecosystems' health are mostly unknown. In recent years, the scientific community started to focus on the possible toxic effects of essential oils and plant extracts towards non-target organisms. As a result, an increasing body of knowledge has emerged. This review describes the current state of the art on the toxic effects that essential oils and plant extracts have towards organisms from different trophic levels, including producers, primary consumers, and secondary consumers. The majority of the studies (76.5%) focuses on the aquatic environment, particularly in aquatic invertebrates (45.1%) with only 23.5% of the studies focusing on the potential toxicity of plant-derived products on terrestrial ecosystems. While some essential oils and extracts have been described to have no toxic effects to the selected organisms or the toxic effects were only observable at high concentrations, others were reported to be toxic at concentrations below the limit set by international regulations, some of them at very low concentrations. In fact, L(E)C50 values as low as 0.0336 mg.L-1, 0.0005 mg.L-1 and 0.0053 mg.L-1 were described for microalgae, crustaceans and fish, respectively. Generally, essential oils exhibit higher toxicity than extracts. However, when the extracts are obtained from plants that are known to produce toxic metabolites, the extracts can be more toxic than essential oils. Overall, and despite being generally considered "eco-friendly" products and safer than they synthetic counterparts, some essential oils and plant extracts are toxic towards non-target organisms. Given the increasing interest from industry on these plant-based products further research using international standardized protocols is mandatory.

Chemical characterization and bioactive potential of Thymus×citriodorus (Pers.) Schreb. preparations for anti-acne applications: Antimicrobial, anti-biofilm, anti-inflammatory and safety profiles.

ETHNOPHARMACOLOGICAL RELEVANCE: Thymus × citriodorus (Pers.) Schreb. is an interspecific hybrid between Thymus pulegioides and Thymus vulgaris, known for its pharmacological activities as diaphoretic, deodorant, antiseptic and disinfectant, the last mostly related with its antimicrobial activity. The folk use of other extracts, as hydrolates, have also been disseminated, as regulators of oily skin with anti-acne effect. AIM OF THE STUDY: We aimed to evaluate the anti-acne potential of two Thymus x citriodorus (TC) preparations, the essential oil (EO) and the hydrolate, to be used as active ingredients for skin applications. Specifically, we intend to validate their anti-acne potential by describing their activity on acne related bacteria, bacterial virulence, anti-oxidant and anti-inflammatory potential, and biocompatibility on inflammatory cells. Additionally, we aimed to report their ecotoxicity under the Globally Harmonized System of Classification and Labelling of Chemicals (GHS), thus focusing not only on the consumer, but also on environmental safety assessment. MATERIALS AND METHODS: Minimum inhibitory concentration (MIC) against C. acnes, S. aureus and S. epidermidis was evaluated. Minimum lethal concentration (MLC) was also determined. The effect on C. acnes biofilm formation and disruption was evaluated with crystal violet staining. Anti-inflammatory activity was investigated on LPS-stimulated mouse macrophages (RAW 264.7), by studying nitric oxide (NO) production (Griess reagent) and cellular biocompatibility through MTT assay. In-vitro NO and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging potential were also evaluated. The ecotoxicity was evaluated using Daphnia magna acute toxicity assays. RESULTS: EO presented direct antimicrobial activity, with visual MICs ranging from 0.06% for S. epidermidis and C. acnes to 0.125% for S. aureus. MLCs were higher than the obtained MICs. Hydrolate revealed visual MIC only for C. acnes. TC essential oil was effective in preventing biofilm formation and disrupting preformed biofilms even at sub-inhibitory concentrations. Hydrolate showed a more modest anti-biofilm effect. Regarding anti-inflammatory activity, TC hydrolate has a higher cellular biocompatibility. Still, both plant preparations were able to inhibit at least 50% of NO production at non-cytotoxic concentrations. Both EO and hydrolate have poor anti-oxidant activities. Regarding the ecotoxicity, TC essential oil was classified under acute 3 category, while the hydrolate has proved to be nontoxic, in accordance to the GHS. CONCLUSIONS: These results support the anti-acne value of different TC preparations for different applications. TC hydrolate by presenting higher biocompatibility, anti-inflammatory potential and the ability to modulate C. acnes virulence, can be advantageous in a product for everyday application. On the other hand, EO by presenting a marked antimicrobial, anti-biofilm and anti-inflammatory activities, still with some cytotoxicity, may be better suited for application in acute flare-ups, for short treatment periods.

Frailty syndrome, biomarkers and environmental factors - A pilot study.

Frailty is an age-related syndrome expected to increase over the next decades. This syndrome has been identified to be the most common condition leading to disability, institutionalisation and death in the elderly. The aim of this pilot study is to investigate a possible link between frailty status, biomarkers and environmental exposures. A group of 71 older adults (≥65 years old) was engaged in this study. The study population was classified as 45.1% robust, 45.1% pre-frail and 9.8% frail. A significant higher prevalence of second-hand smokers was found in the pre-frail group when compared to robust. Furthermore, a higher prevalence of robust individuals was found among those consuming home-produced vegetables and water from well/springs. Significant differences were found between data collected in a lifetime exposure questionnaire (LTEQ) and the levels of genotoxicity endpoints and the mercury levels analysed regarding some exposure-related parameters, namely, smoking habits, intake of home-produced vegetables and the use of pesticides in agriculture. Understanding if the way we live(d) or worked can impact the way we age are important questions to be explored. Data obtained in this pilot study encourage further studies on this matter, exploring the role of exposures history and its impact on health.

A circular toxicity approach to isoprostanes: From markers of oxidative stress, to epidemiological warning systems and agents of aquatic toxicity.

Isoprostanes (IsoPs) are a class of oxidation products naturally formed in vivo that are indicative of endogenous oxidative stress. In individuals with chronic and oxidative stress related diseases, IsoPs are increased to pathological levels. Since they are excreted through urine into sewage systems, IsoPs can be detected in wastewater treatment plants' (WWTPs) effluents and thus can be used to evaluate the health status of a given population. The underlying principle is that higher isoprostanes WWTPs' levels correspond to populations undergoing higher levels of oxidative stress, and thus disease. However, IsoPs are not eliminated by WWTPs and will end up being released into the aquatic environment, where they will be available for uptake by aquatic species. Being bioactive molecules, it has been suggested that IsoPs in the environment may elicit oxidative stress in aquatic organisms. In this context, we have critically reviewed the available data on IsoPs as products and effectors of toxicity, and propose the new concept of "circular toxicity". In general, IsoPs excreted by humans as a consequence of oxidative stress are released into the aquatic environment where they may interact with aquatic organisms and induce the production of more IsoPs. These stress markers, in turn, will also be excreted, increasing the already high levels of stressors in the aquatic environment and thus create an escalating cycle of oxidative stress.

Acute phase proteins response in cats naturally infected by hemotropic mycoplasmas.

Information about the acute phase proteins (APP) response in cats naturally infected with hemoplasmas and in cats co-infected with different species of hemoplasmas is lacking. This study evaluated serum amyloid A (SAA), haptoglobin (Hp) and albumin in 48 cats naturally infected with hemoplasmas, including 25 with Candidatus Mycoplasma haemominutum and 23 co-infected with different hemoplasmas agents; and in 10 healthy control cats. Infected cats had significantly higher Hp and lower albumin than controls. Symptomatic cats had significantly higher SAA and Hp, and lower albumin than asymptomatic animals, and also than controls. Asymptomatic cats had significantly higher Hp than controls. Concentrations of APP were not significantly different between single infected and co-infected cats. According with these results, hemoplasmosis should be considered when alterations in APP are detected in diseased cats with compatible clinical signs. Furthermore, a subclinical infection should be considered in apparently healthy cats from endemic areas with increased Hp.

Levels of TBT and other selected organotin compounds in duplicate diet samples.

Organotin compounds (OTs) are ubiquitous contaminants with a broad range of applications ranging from biocides and pesticides to catalysts for the production of polyurethane foams and silicones. The deleterious effects of some OTs (particularly tributyltin - TBT) upon wildlife and experimental animals are well documented and include endocrine disruption, immunotoxicity, neurotoxicity, genotoxicity and metabolic dysfunction in which obesity is included. However, virtually no data on the current human exposure levels is available. In order to bridge this gap, we quantified for the first time the levels of OTs in duplicate diet samples from members of the University of Aveiro in Portugal. OTs were detected in 32% of the 28 diet samples analyzed, at relatively low levels. TBT and monobutyltin were detected only in two samples and dibutyltin was detectable in three samples. Dioctyltin was quantified in four samples and monooctyltin in three samples. Phenyltins were below the detection limit in all the diet samples analyzed. Overall, for the vast majority of the samples (89%), the estimated daily intakes (EDI) of organotins through food were much lower than the established tolerable daily intakes (TDI). Hence, for the majority of the participants the risk associated with food ingestion is low.

Lead in duplicate diet samples from an academic community.

Lead is a naturally occurring element that with the advent of the industrial era became a serious environmental and public health issue. Leaded gasoline, lead based paints, use of lead in plumbing and water pipes, ceramics with lead-containing glazes and tobacco smoke are potential sources of lead exposure for humans. Despite these multiple sources, food is still considered the most important one for the general non-smoking population. Hence, in the present study, the dietary intake of lead was determined in duplicate diet samples provided by 30 participants working or studying at University of Aveiro, Portugal. Pb was detected in all the analysed samples with values ranging between 0.009 and 0.10mgkg-1 ww which correspond to estimated daily intakes between 0.22 and 3.5µgkg-bw-1day-1. Risk estimations disclose that at least 3.3% and 26.7% of the participants might suffer cardiovascular and nephrotoxic effects, respectively.

Urine protein-to-creatinine concentration ratio in samples collected by means of cystocentesis versus manual compression in cats.

Objective-To compare urine protein-to-creatinine concentration (UPC) ratios in samples collected by means of cystocentesis versus manual compression in cats. Design-Evaluation study. Animals-43 client-owned cats requiring urinalysis. Procedures-In all cats, 5 mL of urine from the midstream phase of micturition was collected by means of manual compression and, subsequently, an additional 5 mL of urine was obtained by means of ultrasound-guided cystocentesis. A complete urinalysis was performed on all samples, and UPC ratios were determined. Results-Cats were classified on the basis of the International Renal Interest Society substaging system as being free from proteinuria (UPC ratio, < 0.2; n = 19) or as having borderline proteinuria (UPC ratio, 0.2 to 0.4; 7) or proteinuria (UPC ratio, > 0.4; 17). None of the cats had postrenal proteinuria. A significant linear correlation was identified between UPC ratios in urine samples obtained by means of manual compression and ratios in samples obtained by means of cystocentesis. For all cats, UPC ratios for samples obtained by the 2 collection methods resulted in classification in the same IRIS substage. Conclusions and Clinical Relevance-Results suggested that collection of a urine sample from the midstream phase of micturition by manual compression would be a reliable alternative to cystocentesis for the determination of UPC ratio in cats, provided that postrenal proteinuria was excluded by means of urine sediment analysis. Once postrenal proteinuria was ruled out, the method used to collect urine samples did not appear to influence the quantification of urine protein concentration.

Metal(loid) levels in biological matrices from human populations exposed to mining contamination--Panasqueira Mine (Portugal).

Mining activities may affect the health of miners and communities living near mining sites, and these health effects may persist even when the mine is abandoned. During mining processes various toxic wastes are produced and released into the surrounding environment, resulting in contamination of air, drinking water, rivers, plants, and soils. In a geochemical sampling campaign undertaken in the Panasqueira Mine area of central Portugal, an anomalous distribution of several metals and arsenic (As) was identified in various environmental media. Several potentially harmful elements, including As, cadmium (Cd), chromium (Cr), manganese (Mn), nickel (Ni), lead (Pb), and selenium (Se), were quantified in blood, urine, hair, and nails (toe and finger) from a group of individuals living near the Panasqueira Mine who were environmentally and occupationally exposed. A group with similar demographic characteristics without known exposure to mining activities was also compared. Genotoxicity was evaluated by means of T-cell receptor (TCR) mutation assay, and percentages of different lymphocyte subsets were selected as immunotoxicity biomarkers. Inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma-atomic emission spectrometry (ICP-AES) analysis showed elevated levels of As, Cd, Cr, Mn, and Pb in all biological samples taken from populations living close to the mine compared to controls. Genotoxic and immunotoxic differences were also observed. The results provide evidence of an elevated potential risk to the health of populations, with environmental and occupational exposures resulting from mining activities. Further, the results emphasize the need to implement preventive measures, remediation, and rehabilitation plans for the region.

A feeding inhibition based prediction of the toxic effect of dissolved metal mixtures upon Echinogammarus marinus (Crustacea: Amphipoda) at field relevant concentrations across a latitudinal gradient.

Risk assessment of metals in the environment is performed mainly with toxicity evaluations on single metals, which is largely inadequate since these substances occur in mixtures. The development of models predicting combined toxic effects on the basis of the concentration-response relationships of individual compounds has emerged as an answer. In the present study, metal effects on post-exposure anorexia (the concept of FdC(50)--concentration causing 50% of feeding inhibition--is implemented) in Echinogammarus marinus, a widely distributed gammarid amphipod, were assessed and compared with modelled ones obtained through the application of the concentration addition (CA) model, which represents a reasonable worst-case scenario for the risk assessment of metal mixtures. Data were validated using in situ experiments performed along a latitudinal gradient (Iceland, Scotland and Portugal) aiming at establishing a geographic profile of autochthonous population susceptibilities to metals. For all of the metals studied concentrations in the water column at exposure sites were in good agreement with feeding inhibition levels. Models gave low to relatively high percentage agreement between predictions and experimental data. Boreal populations demonstrated higher susceptibility to single metals, but not to mixture exposures. Meridional populations denoted lower susceptibilities with higher FdC(50).

Amphipod susceptibility to metals: cautionary tales.

Heavy metals accumulated by aquatic crustaceans in environmental studies are normally investigated using the whole body burden, with little regard paid to uptake in different tissues, to potential gender of life stage differences, or to the influence of nutrition on the test organism. This is likely to give erroneous conclusions for a dose-response relationship within the toxicity test and potentially lead to wrong conclusions for the ecological risks of metals where species may have higher sensitivities with gender and life stage than indicated or that functionally metals may be sequestered into parts of the body so are not bioavailable. This could lead to under-estimation or over-estimation of the toxicity of metals, respectively, inaccuracy of metal budget calculations and evaluation of trophic transfers of metals. This study evaluated the influences of life stage, gender, and a priori nutritional state in the uptake of the metals zinc (an essential micro-nutrient; Zn) and cadmium (a non-essential element; Cd) in the amphipod Echinogammarus marinus. The study showed that life stage, and nutritional stage did significantly influence the uptake and bioaccumulation for both metals, but only Cd showed differential uptake and bioaccumulation with gender. In addition, it was concluded that there was a significant uptake and accumulation of both metals within the exoskeleton of the amphipods, which though adding to the full body burden would add little to toxicity through lack of bioavailability. These results showed that care should be taken when interpreting results from tests normally preformed on such test organisms.

Amphipod intersex, metals and latitude: a perspective.

Intersexuality has been widely reported in crustaceans with several mechanisms being directly or indirectly held responsible for its occurrence, amongst which pollution. No mechanistic relationship between metals and intersex has ever been established. Also the incidence of intersex in populations of the same invertebrate species in a latitudinal gradient has never been studied so far. Three populations (Iceland, Scotland and Portugal) of the amphipod Echinogammarus marinus were scrutinized. Intersex females from Iceland registered the highest fecundity loss. Only in Scottish samples females with two genital papillae and males with only one genital papillae were observed. Nevertheless, water, biota and sediment samples pointed to equivalent metal levels, and in consonance the prevalence of intersex was not significantly different between locations. An unequivocal relationship between metal presence and intersex induction cannot be presented, but our results advocate the potential role of metals as a direct cause of intersexuality in E. marinus.