First evidence of environmental bioaccumulation of pharmaceuticals on adult native anurans (Rhinella arenarum) from Argentina.

The presence of pharmaceutically active compounds (PhACs) in surface water is well known, whereas their natural occurrence in biota is much less explored. The aim of this work was to evaluate the bioaccumulation of PhACs in adult toads of the neotropical species Rhinella arenarum. Three sites were selected in Buenos Aires (Argentina): a reference site (Site 1), a site with direct discharge from a secondary wastewater treatment plant (WWTP) (Site 2) and a site 300 m downstream of the WWTP discharge (Site 3). Surface water samples, as well as muscle, liver and fat bodies of toads were collected, extracted and analyzed by LC-MS/MS. Highly significant differences in total PhACs concentration in surface water (p < 0.005) were detected between Site 2 and the other sites. These concentrations ranged from 0.37 to 52.46 ng/L at Site 1, 0.71-6950.37 ng/L at Site 2, and 0.12-75.45 ng/L at Site 3. In general, bioaccumulation of PhACs in toad tissues was similar between sites and tissues of each site. The highest concentrations were detected in the muscle of toads from Site 3 (1.06-87.24 ng/g dw), followed by liver (1.77-38.10 ng/g dw) and fat bodies (0.68-20.59 ng/g dw) from Site 1. Ibuprofen (6950 ng/L), acetaminophen (3277 ng/L) and valsartan (2504 ng/L) were the compounds with the highest concentrations in surface water from Site 2, whereas acetaminophen (87.2 ng/g dw, muscle from Site 3), desloratadine (38.1 ng/g dw, liver from Site 1), and phenazone (25.9 ng/g dw, liver from Site 1) were the ones that showed the highest concentrations in biota. This is the first time a field study has examined the environmental bioaccumulation of PhACs in anurans, demonstrating their potential for monitoring the status of natural ecosystems.

Realistic scenarios of pesticide exposure alters multiple biomarkers in BOANA PULCHELLA (ANURA) Adult Frogs.

Imazethapyr, a post-emergent herbicide used in worldwide soybean and corn crops, induces genetic and biochemical alterations in aquatic vertebrates. This study examined the relationship between biomarkers at different organization levels and imazethapyr real-life route exposure in Boana pulchella adults. Frogs were exposed to imazethapyr-based formulation Pivot® H (10.59%) at concentrations representing possible acute routes: field runoff (S1:10 mg.L-1), exposure after direct foliar application (S2:100 mg.L-1) and during direct foliar application (S3:1000 mg.L-1). Post-exposure, endpoints levels were evaluated: organism alterations, biochemical activities and cytogenetic assays. Forty-eight hours post-exposure, antioxidant enzymes decrease, micronuclei induction and DNA damage were observed in all scenarios, while cholinesterase activity increase and body condition reduction were observed in frog-exposed to S3. Ninety-six hours post-exposure, frogs showed glutathione-S-transferase inhibition in S1, micronuclei induction in S2 and S3, and DNA-damage increase in S3. Herbicides routes of exposures in real-life could indicate that authorized applications have a risk to amphibian populations.

Multiple Level Effects of Imazethapyr on Leptodactylus latinasus (Anura) Adult Frogs.

Imazethapyr is an herbicide that is used in a variety of crops worldwide, including soybean and corn. The aim of the present study was to evaluate the biomarkers responses of adult Leptodactylus latinasus exposed to the formulation Pivot® H (10.59% imazethapyr) in the laboratory at concentrations and under conditions that simulate two potential field exposure scenarios: an immersion in field runoff (Scenario 1: 10 mg/L) and a direct exposure to the droplets emitted by spray noozles (Scenario 2: 1000 mg/L). In both scenarios, the experimental procedure involved completely immersing the frogs over a period of 15 s. Different endpoints were evaluated at several ecotoxicological levels 48 and 96 h after the herbicide exposure. These included individual (biometric indices and behavior alterations), histological (liver pigments and lesions), biochemical (catalase, glutathione system and cholinesterase activities) and genotoxic effects (micronuclei induction and nuclear abnormalities). Forty-eight hours after imazethapyr exposure, frogs submitted to Scenario 1 presented an inhibition of liver glutathione-S-transferase activity, whereas histological alterations and increased hepatic cholinesterase levels were observed in frogs exposed under Scenario 2. Ninety-six hours after exposure to the imazethapyr formulation, frogs from the Scenario 1 treatment presented a decrease in liver melanin and hemosiderin, increased hepatic catalase activity and micronuclei induction. For their part, frogs exposed to Scenario 2 presented a decrease in the hepatosomatic index, an increase in liver alterations, melanin reduction and micronuclei induction. The multivariate analysis enables correlations to be made between biomarkers of different organizational level in exposed anurans. Our result indicates that real exposure to imazethapyr formulations under field conditions may pose a risk to Leptodactylus latinasus populations living in the agroecosystems.

Lethal and sublethal effects of the pirimicarb-based formulation Aficida® on Boana pulchella (Duméril and Bibron, 1841) tadpoles (Anura, Hylidae).

Acute lethal and sublethal toxicity of the pirimicarb-based commercial formulation Aficida® were evaluated on Boana pulchella tadpoles. Whereas mortality was used as end point for lethality, frequency of micronuclei and other nuclear abnormalities as well as alterations in the frequency of erythroblasts in circulating blood as biomarkers for genotoxicity and cytotoxicity, respectively. Swimming, growth, developmental and morphological abnormalities were also employed as sublethal end points. Results show that the species is within the 13th percentile of the distribution of acute sensitivity of species to pirimicarb for aquatic vertebrates. Results revealed values of 23.78 and 101.45mg/L pirimicarb as LC5096h for GS25 and GS36 tadpoles, respectively. The most evident effects were related with the swimming activity with NOEC and LOEC values within the 0.005-0.39mg/L pirimicarb concentration range. Aficida® induced DNA damage at the chromosomal level by increasing micronuclei frequency and other nuclear abnormalities, i.e., lobbed and notched nuclei and binucleated cells. Cellular cytotoxicity was found after Aficida® treatment. The presence of abdominal oedemas in exposed organisms and thus flotation response of organisms could be proposed as a new sensitive exposure parameter. The multiple end point assessment approach used allowed a complete understanding the multi level of effects occurring by exposure to pirimicarb, at least in B. pulchella.

Effects of the Fruit Ripening Stage on Antioxidant Capacity, Total Phenolics, and Polyphenolic Composition of Crude Palm Oil from Interspecific Hybrid Elaeis oleifera × Elaeis guineensis.

In the present study, we assessed for the first time the changes in the antioxidant capacity, total phenolic content, and polyphenolic composition of interspecific hybrid palm oil extracted from Elaeis oleifera × Elaeis guineensis (O × G, Coari × La Mé cultivar) during the fruit ripening process 18, 20, 22, and 24 weeks after anthesis. A progressive decrease (p < 0.05) of phenolic content occurred during fruit development together with marked changes in polyphenol profiles. Significant negative correlations were established between antioxidant activity measured by TEAC (R = -0.954; p < 0.05) and ORAC (R = -0.745; p < 0.05) and the fruit ripening stage, while a positive correlation between total phenolic content was found using either the TEAC assay or the ORAC assay. The highest DPPH radical scavenging activity was also obtained with oils extracted at 18 WAA. These results highlight that O × G fruits of early ripeness represent a better source of phenolic compounds and may provide extracts with higher antioxidant activities when hybrid palm oil is aimed to be used as a functional ingredient for the development of food or food products with antioxidant properties.

Toxic and genotoxic effects of the imazethapyr-based herbicide formulation Pivot H® on montevideo tree frog Hypsiboas pulchellus tadpoles (Anura, Hylidae).

Acute lethal and sublethal toxicity of the imidazolinone imazethapyr (IMZT)-based commercial formulation herbicide Pivot H® (10.59% IMZT) was evaluated on Hypsiboas pulchellus tadpoles. Whereas mortality was used as the end point for lethality, frequency of micronuclei (MNs) and other nuclear abnormalities as well as DNA single-strand breaks evaluated by the single cell gel electrophoresis assay were employed to test genotoxicity. Behavioral, growth, developmental, and morphological abnormalities were also employed as sublethal end points. Mortality studies revealed equivalent LC50 (96h) values of 1.49mg/L (confidence limit, 1.09-1.63) and 1.55mg/L (confidence limit, 1.51-1.60) IMZT for Gosner stage (GS) 25 and GS36, respectively. Behavioral changes, i.e., irregular swimming and immobility, as well as a decreased frequency of keratodonts were observed. The herbicide increased the frequency of MNs in circulating erythrocytes of tadpoles exposed for 48h to the highest concentration assayed (1.17mg/L). However, regardless of the concentration of the herbicide assayed, an enhanced frequency of MNs was observed in tadpoles exposed for 96h. The herbicide was able to induce other nuclear abnormalities, i.e., blebbed and notched nuclei, only when tadpoles were exposed for 96h. In addition, we observed that exposure to IMZT within the 0.39-1.17mg/L range increased the genetic damage index in treatments lasting for both 48 and 96h. This study represents the first evidence of acute lethal and sublethal effects exerted by IMZT on amphibians. Finally, our findings highlight the properties of this herbicide that jeopardize nontarget living species exposed to IMZT.

Punica granatum cv. Dente di Cavallo seed ethanolic extract: antioxidant and antiproliferative activities.

This paper aims to provide a solid base for the utilisation of pomegranate whole seed ethanolic extract (PSEE) as a nutraceutical/functional food ingredient. PSEE was tested for its antioxidant and antiproliferative activities against different human cancer cell lines. Bioactive lipid compounds were identified by studying the PSEE lipid portion. PSEE exhibited a protection of lipid peroxidation threefold higher than a positive control. PSEE showed a promising antiproliferative activity against hormone dependent prostate carcinoma LNCaP, with an IC50 value 3 times lower than the positive control vinblastine, and against human breast cancer cell lines (IC50=9.6 µg/ml). PSEE contained lipid bioactive compounds, such as neutral lipids, consisting of 72.8% punicic acid, glycolipids and phospholipids rich in essential fatty acids (α-linoleic and α-linolenic acids). Due the presence of bioactive compounds and the remarkable antiproliferative activity, the use of PSEE as a value-added ingredient in formulations of products aimed to prevent diseases, especially cancer, could be promoted.

Acrylamide levels in selected Colombian foods.

Acrylamide (AA) levels in conventional (n = 112) and traditional (n = 43) Colombian foods were analysed by gas chromatography with mass spectrometry (GC/MS) detection. Samples included: infant powdered formula, coffee and chocolate powders, corn snacks, bakery products and tuber-, meat- and vegetable-based foods. There was a wide variability in AA levels among different foods and within different brands of the same food, especially for coffee powder, breakfast cereals biscuits and French fries samples. Among the conventional foods tested, the highest mean AA value was found in bakery products, such as biscuit (1104 µg kg(-1)) and wafer (1449 µg kg(-1)), followed by potato chips (916 µg kg(-1)). On the other hand, among the traditional foods, higher AA amounts were detected in fried platano (2813 µg kg(-1)) and yuca (3755 µg kg(-1)) compared to other products. Interestingly, the arepa, a traditional Colombian bakery product made with corn flour, showed a lower AA content (< 75 µg kg(-1)) when compared with similar bakery products tested, such as soft bread (102-594 µg kg(-1)), which is a made with wheat flour.

The genotoxic effects of the imidacloprid-based insecticide formulation Glacoxan Imida on Montevideo tree frog Hypsiboas pulchellus tadpoles (Anura, Hylidae).

The neonicotinoid insecticide imidacloprid (IMI) affects the insect central nervous system and is successfully applied to control pests for a variety of agricultural crops. In the current study, acute toxicity and genotoxicity of the IMI-containing commercial formulation insecticide Glacoxan Imida (35 percent IMI) was evaluated on Hypsiboas pulchellus (Anura: Hylidae) tadpoles exposed under laboratory conditions. A lethal effect was evaluated as the end point for lethality, whereas micronucleus (MN) frequency and DNA single-strand breaks evaluated by the single cell gel electrophoresis (SCGE) assay were employed as end points for genotoxicity. Sublethal end points were assayed within the 12.5-37.5mg/L IMI concentration range. Experiments were performed on tadpoles at stage 36 (range, 35-37) according to the classification proposed by Gosner. Lethality studies revealed an LC50 96h value of 52.622mg/L IMI. Increased frequency of MNs was only observed when 25.0mg/L was assayed for 96h, whereas no other nuclear abnormalities were induced. Increase of the genetic damage index was observed at 48h of treatment within the 12.5-37.5mg/L concentration range, whereas an increased frequency of DNA damage was observed only in tadpoles treated with 37.5mg/L IMI for 96h. This study represents the first evidence of the acute lethal and genotoxic effects exerted by IMI on tadpoles of an amphibian species native to Argentina under laboratory conditions.

Abnormalities in amphibian populations inhabiting agroecosystems in Northeastern Buenos Aires Province, Argentina.

The occurrence of abnormalities in amphibians has been reported in many populations, and its increase could be related to environmental pollution and habitat degradation. We evaluated the type and prevalence of abnormalities in 5 amphibian populations from agroecosystems with different degrees of agricultural disturbance (cultivated and reference areas). We detected 9 types of abnormalities, of which the most frequent were those occurring in limbs. The observed prevalence of abnormality in assessed populations from cultivated and reference areas was as follows: Rhinella fernandezae (37.1 and 10.2%, respectively), Leptodactylus latrans adults (28.1 and 9.2%) and juveniles (32.9 and 15.3%), and Hypsiboas pulchellus (11.6 and 2.8%). Scinax granulatus populations did not show abnormalities. Pseudis minuta, which was only detected in the reference area, exhibited a prevalence of 13.3%. For R. fernandezae, L. latrans, and H. pulchellus, generalized linear mixed models showed that prevalence of abnormalities was significantly higher (p < 0.05) in cultivated than in reference areas. L. latrans juveniles were more vulnerable to abnormalities than adults (p < 0.05). The presence of abnormalities in some species inhabiting different agroecosystems suggests that environmental stress factors might be responsible for their occurrence. While we detected pesticides (endosulfan, cypermethrin, and chlorpyrifos) and lower dissolved oxygen levels in ponds of the cultivated area, no data are currently available on how other factors, such as injuries from predators and parasite infections, vary by land use. Further research will be necessary to evaluate possible causes of abnormalities detected in the present study mainly in the context of factor interactions.

Grados de depresión en padres de niños con cáncer / Depression grades in parents which have cancer children

Es bien conocido que las enfermedades somáticas padecidas por los pacientes, afectan el estado psicológico de las personas con quienes estos guardan un vínculo cercano. Intentamos corroborar este hecho aplicándoles test de Hamilton a 58 padres de pacientes, hospitalizados en las alas de oncología pediátrica de diversos centros de salud de Caracas. Se observó una prevalencia de depresión en el 90% de los padres y además pudimos ver que los grados de depresión variaban de acuerdo a ciertos determinantes como el tiempo de desarrollo de la enfermedad y el ambiente del centro hospitalario. Siendo por ende necesario brindar atención psicológica especializada a los acompañantes de los pacientes en los distintos centros de salud

It is well known that the suffering of an illness can affect not only the patient, but also every person that relates intimately to him. We attempted the verification of this statement by applying Hamilton tests to 58 parents of patients which were hospitalized in the several pediatric oncology-specialized health care centers in Caracas. The outcome was a predominance of depression in 90% of the parents; in addition, observed how the different degrees of depression could fluctuate according to certain factors like: Development time of the illness and the hospital’s environment. It is necessary to provide specialized psychological care to the relatives or any companions of the patients hospitalized in health care centers

Acute and chronic effects of Cr(VI) on Hypsiboas pulchellus embryos and tadpoles.

In the last few years there has been great concern about declines in the abundance of several species of amphibians around the world. Among amphibians, anurans have a biphasic life cycle, with aquatic tadpoles and generally terrestrial adults, and they have an extremely permeable skin, making them excellent indicators of the health of the environment. A number of different causes have been suggested for the global decline of anurans, the pollution of their habitat by chemical stressors being considered one of the major factors. Among chemical stressors, heavy metals are known for their high toxicity at very low concentrations. This study assessed short- (96 h, 'acute') and long-term (1272 h, 'chronic') exposure to Cr(VI) at lethal (3 to 90 mg 1(-1)) and sublethal concentrations (0.001 to 12 mg 1(-1)) on Hypsiboaspulchellus (previously called Hyla pulchella; see Faivovich et al. 2005) tadpoles (Fam. Hylidae) from central eastern Argentina. Fertilized eggs collected from a clean pond near La Plata (Buenos Aires Province) were used for acute and chronic toxicity testing. Assays were done under controlled laboratory conditions. Results of chronic exposure were used to assess the effect of factors such as toxicant concentration and age of organisms at the beginning of exposure on the response variables (growth, development and survival until metamorphosis). Results indicated a higher sensitivity to Cr(VI) of individuals first exposed as tadpoles than those first exposed as embryos during acute and chronic exposure. Exposure to the highest sublethal concentrations (6 to 12 mg 1(-1)) of the toxicant showed early inhibitory effects on growth of all treated organisms compensated at longer exposure periods with an increase in the growth rate compared to the control groups.