Heavy metals and TPH effects on microbial abundance and diversity in two estuarine areas of the southern-central coast of São Paulo State, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of soil texture in the recovery of Toxocara canis eggs by a flotation method

In epidemiological surveys, the evaluation of soil contamination by Toxocara canis eggs requires a quick and easy method for the isolation of parasite eggs from soil samples. The efficiency of flotation methods is influenced by sample size, soil texture, degree of soil contamination, pretreatment, flotation solutions and time of flotation. This investigation was designed to evaluate the influence of soil texture in the recovery of T. canis eggs with the centrifugal flotation technique of Dada (Dada, B.J.O., 1979. A new technique for the recovery of Toxocara eggs from soil. J. Helminthol., 53: 141-144). Four types of soil (clay silt, sandy, silty clay and sand) were artificially contaminated with T. canis eggs (200 eggs per gram). Zinc sulphate (specific gravity 1.20) and sodium dichromate (specific gravity 1.35) were used as flotation solutions. Twenty replicated examinations were performed for each type of soil and flotation solution. There was a statistically significant difference in the results depending on soil type. The highest recovery percentages were observed in soils rich in sand (62.5% for sand and 38.0% for sandy soil). Differences were also observed with different flotation solutions. Sodium dichromate solution was more efficient for recovering T. canis eggs, regardless of the soil texture. © 1994.

Avaliação do processo de contaminação e remediação de água subterrânea por hidrocarbonetos

Pós-graduação em Engenharia Civil e Ambiental - FEB

Avaliação de contaminação com óleo diesel por meio dos métodos de Eletrorresistividade e Polarização Induzida em Santa Ernestina-SP

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Microbiological quality and safety of minimally processed vegetables marketed in Campinas, SP-Brazil, as assessed by traditional and alternative methods

In this study, a total of 172 samples of minimally processed vegetables (MPV) were collected from supermarkets in the city of Campinas, Brazil. The MPV were analyzed using traditional and/or alternative methods for total aerobic mesophilic bacteria, total coliforms, Escherichia coil, coagulase positive staphylococci, Salmonella and Listeria monocytogenes. All the MPV analyzed presented populations of aerobic mesophilic microorganisms and total coliforms were >4 log(10) CFU/g and 1.0-3.4 log(10) CFU/g, respectively. E. coil was enumerated in only 10 samples out of 172 collected, while none of the 172 samples of MPV presented contamination by coagulase positive Staphylococcus (<10(1) CFU/g). Among the four methods used for detection of Salmonella in MPV (Vidas, 1,2 Test, Reveal, and Traditional), when Reveal was used a total of 29 positive samples were reported. For L monocytogenes, the four methods tested (Vidas, Vip, Reveal, and traditional) performed similarly. The presence of Salmonella and L monocytogenes in MPV was confirmed in one (watercress) and two samples (watercress and escarole), respectively. In conclusion, it has been observed that the microbiological quality of MPV commercialized in Campinas is generally satisfactory. Besides, the choice of microbiological method should be based not only on resource and time issues, but also on parameters such as sensitivity and specificity for the specific foods under ahalysis. (C) 2012 Elsevier Ltd. All rights reserved.

Herbicides in the upper Poxim River, Sergipe, and the risk of contamination of water resources

Increased agricultural activity in watershed areas has been causing concern over contamination by herbicides in agricultural areas. The problem becomes more important when contamination can affect water for human consumption, as happens with water from the Poxim river, which supplies the city of Aracaju, capital of the State of Sergipe. The aim of this study was to evaluate the risk of contamination by herbicides to both surface and groundwater in the upper sub-basin of the Poxim River, and to detect the presence of the active ingredients Diuron and Ametrine up-river from the sugar-cane plantations. Risk analysis was carried out using criteria from the Environmental Protection Agency (EPA), the GUS index, and the GOSS method. It was observed that several active ingredients are at risk of leaching, demonstrating the importance of monitoring the river to control both the quality of water and the frequency and volume of herbicides used in the region. Based on the results, monitoring was carried out bi-monthly from July 2009 to July 2010 at two sampling points. Water samples were analyzed in the laboratory, where the presence of Diuron and Ametrine was noted. Water quality in the Sub-basin of the Rio Poxim is being influenced by the use of herbicides in the region. There was an increase in herbicide concentration in the surface water during the rainy season, possibly caused by soil runoff.

Stable isotope fractionation of selenium by biomethylation in soil

Selen ist in geringen Mengen ein essentielles Nährelement, das aber in höheren Gehalten toxisch wird. Der Se-Kreislauf in der Umwelt ist eng mit Redoxreaktionen wie der Reduktion von Se-Oxyanionen zu Methylselenid verknüpft. Flüchtige Methylselenide sind weit verbreitet und stellen einen wichtigen Se-Fluss in der Umwelt dar. Das übergeordnete Ziel meiner Dissertation war, die Stabilisotopenfraktionierung von Se durch Biomethylierung verschiedener oxidierter Se-Spezies (Se[IV] und Se[VI]) im Boden aufzuklären. Zunächst wurde eine Methode entwickelt, die es erlaubte flüchte Methylselenide quantitativ zurückzuhalten. Es zeigte sich, dass alkalische Peroxid-Lösung dafür geeignet war. Mit alkalischer Peroxid-Lösung wurde eine Wiederfindung von 95,6 ± Standardabweichung 5,4% in Verflüchtigungsexperimenten mit Methylselenid-Standards erreicht. Bei Einsatz von alkalischer Peroxid-Lösung in geschlossenen Mikrokosmos-Experimenten kam es zu keinen Se-Verlusten und ausgeglichenen Se-Isotopenbilanzen. Die massengewichteten δ82/76Se-Werte lagen für Se(IV) und Se(VI) am Ende der Mikrokosmos-Inkubationen bei -0,31 ± 0,05‰ (n = 3) und -0,76 ± 0,07‰ (n = 3) verglichen mit -0,20 ± 0,05‰ und -0,69 ± 0,07‰ im jeweils zugegebenen Se. Im zweiten Teil meiner Dissertation wurde die Pilzart Alternaria alternata mit Se(VI) und Se(IV) in geschlossenen Mikrokosmen für 11-15 und Se(IV) zusätzlich für 3-5 Tage bei 30°C inkubiert. In 11-15 Tagen wurden 2,9-11% des Se(VI) und 21-29% des Se(IV) und in 3-5 Tagen, 3-5% des Se(IV) methyliert. Die anfänglichen δ82/76Se-Werte von Se(VI) und Se(IV) lagen bei -0,69 ± 0,07‰, und -0,20 ± 0,05‰. Die δ82/76Se-Werte der Methylselenide unterschieden sich nach 11-15 Tagen Inkubation signifikant zwischen Se(VI) (-3,97 bis -3,25 ‰) und Se(IV) (-1,44 bis -0,16‰) als Quellen. Die δ82/76Se-Werte der Methylselenide zeigen also die Quellen der Biomethylierung von Se an. Die kürzere Inkubation von Se(IV) für 3-5 Tage führte zu einer ausgeprägten Se-Isotopenfraktonierung von mindestens -6‰, bevor ein Fließgleichgewicht erreicht wurde. Im dritten Teil bestimmte ich die Bindungsformen von Se mit drei operativ definierten sequentiellen Extraktionen und die δ82/76S-Werte des gesamten Selens in zehn urbanen Oberböden mit 0,09-0,52 mg/kg Se, die fünf verschiedene Landnutzungstypen repräsentierten (Überschwemmungsgrünland, Garten, Park, Straßenrand und Wald). Nur ein kleiner Teil des Seleniums lag in austauschbarer und damit direkt bioverfügbarer und in residualer, wenig reaktiver Form vor. Das meiste Se war an die organische Substanz und Fe-(Hydr-)Oxide gebunden (42-77% des gesamten Selens). Der mittlere δ82/76Se-Wert des gesamten Selens in den Oberböden lag mit -0,03 ± 0,38‰ nahe beim Mittelwert der gesamten Erde. Geringfügig niedrigere Se-Isotopensignale von -0,59 bis -0,35‰ v.a. in Waldböden und geringfügig höhere von 0,26 to 0,45‰ in Überschwemmungsgrünland wurden vermutlich durch Boden-Pflanze-Recycling und Se-Kontaminationen durch das Flusswasser verursacht. Der vierte Teil umfasste ein “Natural Attenuation”-Experiment und Mikrokosmos-Inkubationen von Bodenproben mit A. alternata. Die Equilibrierung von zum Boden gegebenem Se(IV) und Se(VI) für drei Tage führte zu abnehmenden wasserlöslichen Se-Gehalten um 32-44% bzw. 8-14, die mit kleinen Isotopenfraktionierung (ε = -0,045 bis -0,12 ‰ and -0,05 to -0,07‰ verbunden waren. In zwei der inkubierten Böden mit mäßig sauren pH-Werten wurden zwischen 9,1 und 30% des zugefügten Se(IV) und 1,7% des zugefügten Se(VI) methyliert während in einem stark sauren Boden keine Methylierung auftrat. Das aus Se(IV) entstandene Methylselenid war deutlich gegenüber dem zugegebenen Se-Standard (0,20‰) an 82Se verarmt (δ82/76Se = -3,3 bis -4,5‰). Meine Ergebnisse zeigen, dass die stabilen Isotopenverhältnisse von Se neue Einblicke in Se-Transformationsprozesse erlauben.rn

Homeopathic Preparations of Quartz, Sulfur and Copper Sulfate Assessed by UV-Spectroscopy

Homeopathic preparations are used in homeopathy and anthroposophic medicine. Although there is evidence of effectiveness in several clinical studies, including double-blinded randomized controlled trials, their nature and mode of action could not be explained with current scientific approaches yet. Several physical methods have already been applied to investigate homeopathic preparations but it is yet unclear which methods are best suited to identify characteristic physicochemical properties of homeopathic preparations. The aim of this study was to investigate homeopathic preparations with UV-spectroscopy. In a blinded, randomized, controlled experiment homeopathic preparations of copper sulfate (CuSO(4); 11c-30c), quartz (SiO(2); 10c-30c, i.e., centesimal dilution steps) and sulfur (S; 11×-30×, i.e., decimal dilution steps) and controls (one-time succussed diluent) were investigated using UV-spectroscopy and tested for contamination by inductively coupled plasma mass spectrometry (ICP-MS). The UV transmission for homeopathic preparations of CuSO(4) preparations was significantly lower than in controls. The transmission seemed to be also lower for both SiO(2) and S, but not significant. The mean effect size (95% confidence interval) was similar for the homeopathic preparations: CuSO(4) (pooled data) 0.0544% (0.0260-0.0827%), SiO(2) 0.0323% (-0.0064% to 0.0710%) and S 0.0281% (-0.0520% to 0.1082%). UV transmission values of homeopathic preparations had a significantly higher variability compared to controls. In none of the samples the concentration of any element analyzed by ICP-MS exceeded 100 ppb. Lower transmission of UV light may indicate that homeopathic preparations are less structured or more dynamic than their succussed pure solvent.

Reconstruction of in-situ porosity and porewater compositions of low-permeability crystalline rocks: Magnitude of artefacts induced by drilling and sample recovery

Geological site characterisation programmes typically rely on drill cores for direct information on subsurface rocks. However, porosity, transport properties and porewater composition measured on drill cores can deviate from in-situ values due to two main artefacts caused by drilling and sample recovery: (1) mechanical disruption that increases porosity and (2) contamination of the porewater by drilling fluid. We investigated the effect and magnitude of these perturbations on large drill core samples (12–20 cm long, 5 cmdiameter) of high-grade, granitic gneisses obtained from 350 to 600 m depth in a borehole on Olkiluoto Island (SW Finland). The drilling fluid was traced with sodium–iodide. By combining out-diffusion experiments, gravimetry, UV-microscopy and iodide mass balance calculations, we successfully quantified the magnitudes of the artefacts: 2–6% increase in porosity relative to the bulk connected porosity and 0.9 to 8.9 vol.% contamination by drilling fluid. The spatial distribution of the drilling-induced perturbations was revealed by numerical simulations of 2D diffusion matched to the experimental data. This showed that the rims of the samples have a mechanically disrupted zone 0.04 to 0.22 cm wide, characterised by faster transport properties compared to the undisturbed centre (1.8 to 7.7 times higher pore diffusion coefficient). Chemical contamination was shown to affect an even wider zone in all samples, ranging from 0.15 to 0.60 cm, inwhich iodide enrichmentwas up to 180 mg/kgwater, compared to 0.5 mg/kgwater in the uncontaminated centre. For all samples in the present case study, it turned out that the magnitude of the artefacts caused by drilling and sample recovery is so small that no correction is required for their effects. Therefore, the standard laboratory measurements of porosity, transport properties and porewater composition can be taken as valid in-situ estimates. However, it is clear that the magnitudes strongly depend on site- and drilling-specific factors and therefore our results cannot be transferred simply to other locations. We recommend the approach presented in this study as a route to obtain reliable values in future drilling campaigns aimed at characterising in-situ bedrock properties.

Measurement of isotopic uranium in water for compliance monitoring by liquid scintillation counting with alpha/beta discrimination

A simple and inexpensive method is described for analysis of uranium (U) activity and mass in water by liquid scintillation counting using $\alpha$/$\beta$ discrimination. This method appears to offer a solution to the need for an inexpensive protocol for monitoring U activity and mass simultaneously and an alternative to the potential inaccuracy involved when depending on the mass-to-activity conversion factor or activity screen.^ U is extracted virtually quantitatively into 20 ml extractive scintillator from a 1-$\ell$ aliquot of water acidified to less than pH 2. After phase separation, the sample is counted for a 20-minute screening count with a minimum detection level of 0.27 pCi $\ell\sp{-1}$. $\alpha$-particle emissions from the extracted U are counted with close to 100% efficiency with a Beckman LS6000 LL liquid scintillation counter equipped with pulse-shape discrimination electronics. Samples with activities higher than 10 pCi $\ell\sp-1$ are recounted for 500-1000 minutes for isotopic analysis. Isotopic analysis uses events that are automatically stored in spectral files and transferred to a computer during assay. The data can be transferred to a commercially available spreadsheet and retrieved for examination or data manipulation. Values for three readily observable spectral features can be rapidly identified by data examination and substituted into a simple formula to obtain $\sp{234}$U/$\sp{238}$U ratio for most samples. U mass is calculated by substituting the isotopic ratio value into a simple equation.^ The utility of this method for the proposed compliance monitoring of U in public drinking water supplies was field tested with a survey of drinking water from Texas supplies that had previously been known to contain elevated levels of gross $\alpha$ activity. U concentrations in 32 samples from 27 drinking water supplies ranged from 0.26 to 65.5 pCi $\ell\sp{-1}$, with seven samples exceeding the proposed Maximum Contaminant Level of 20 $\mu$g $\ell\sp{-1}$. Four exceeded the proposed activity screening level of 30 pCi $\ell\sp{-1}$. Isotopic ratios ranged from 0.87 to 41.8, while one sample contained $\sp{234}$U activity of 34.6 pCi $\ell\sp{-1}$ in the complete absence of its parent, $\sp{238}$U. U mass in the samples with elevated activity ranged from 0.0 to 103 $\mu$g $\ell\sp{-1}$. A limited test of screening surface and groundwaters for contamination by U from waste sites and natural processes was also successful. ^

Defects in embryonic hindbrain development and fetal resorption resulting from vitamin A deficiency in the rat are prevented by feeding pharmacological levels of all-trans-retinoic acid

Vitamin A is required for reproduction and normal embryonic development. We have determined that all-trans-retinoic acid (atRA) can support development of the mammalian embryo to parturition in vitamin A-deficient (VAD) rats. At embryonic day (E) 0.5, VAD dams were fed purified diets containing either 12 μg of atRA per g of diet (230 μg per rat per day) or 250 μg of atRA per g of diet (4.5 mg per rat per day) or were fed the purified diet supplemented with a source of retinol (100 units of retinyl palmitate per day). An additional group was fed both 250 μg of atRA per g of diet in combination with retinyl palmitate. Embryonic survival to E12.5 was similar for all groups. However, embryonic development in the group fed 12 μg of atRA per g of diet was grossly abnormal. The most notable defects were in the region of the hindbrain, which included a loss of posterior cranial nerves (IX, X, XI, and XII) and postotic pharyngeal arches as well as the presence of ectopic otic vesicles and a swollen anterior cardinal vein. All embryonic abnormalities at E12.5 were prevented by feeding pharmacological amounts of atRA (250 μg/g diet) or by supplementation with retinyl palmitate. Embryos from VAD dams receiving 12 μg of atRA per g of diet were resorbed by E18.5, whereas those in the group fed 250 μg of atRA per g of diet survived to parturition but died shortly thereafter. Equivalent results were obtained by using commercial grade atRA or atRA that had been purified to eliminate any potential contamination by neutral retinoids, such as retinol. Thus, 250 μg of atRA per g of diet fed to VAD dams (≈4.5 mg per rat per day) can prevent the death of embryos at midgestation and prevents the early embryonic abnormalities that arise when VAD dams are fed insufficient amounts of atRA.

Preliminary evaluation of the occurrence of herbicides and PAHs in the Wet Tropics region of the Great Barrier Reef, Australia, using passive samplers

The proximity of the Great Barrier Reef (GBR) Marine Park to areas of intensive agriculture and increasing urbanisation places the park under potential threat of contamination by land-based pollutants. Passive samplers were deployed at inshore reef and river mouth sites in the Wet Tropics region of the GBR during a dry and a wet season to measure levels of land-based organic pollutants in this environment. Two types of passive sampling devices were deployed: (i) a polar sampler, which can be used to monitor polar herbicides and (ii) semipermeable membrane devices (SPMDs) which sequester more hydrophobic compounds (e.g. PAHs, chlorpyrifos). Herbicides (diuron, simazine, atrazine, hexazinone and/or flumeturon) were detected at low concentrations (ng L-1) at all sites sampled and in both seasons. Chlorpyrifos was not detected while PAHs were present in SPMDs at levels below limits of detection. The results show that the GBR environment does contain low levels of organic pollutants and that passive sampling provides a sensitive monitoring tool for measuring waterborne organic pollutants. (c) 2005 Elsevier Ltd. All rights reserved.

Protein-mediated isolation of plasmid DNA by a zinc finger-glutathione S-transferase affinity linker

The sequence-specific affinity chromatographic isolation of plasmid DNA from crude lysates of E. coli DH5α fermentations is addressed. A zinc finger-GST fusion protein that binds a synthetic oligonucleotide cassette containing the appropriate DNA recognition sequence is described. This cassette was inserted into the Smal site of pUC19 to enable the affinity isolation of the plasmid. It is shown that zinc finger-GST fusion proteins can bind both their DNA recognition sequence and a glutathione-derivatized solid support simultaneously. Furthermore, a simple procedure for the isolation of such plasmids from clarified cell lysates is demonstrated. Cell lysates were clarified by cross-flow Dean vortex microfiltration, and the permeate was incubated with zinc finger-GST fusion protein. The resulting complex was adsorbed directly onto glutathione-Sepharose. Analysis of the glutathione-eluted complex showed that plasmid DNA had been recovered, largely free from contamination by genomic DNA or bacterial cell proteins. © 2002 Wiley Periodicals, Inc.

Is Rapid Adaptation to New Environments Fueled by Old Mutations? A Case Study of Copper Tolerance on Mimulus guttatus

Rapid adaptation and tolerance is a phenomenon experienced by a variety of organisms typically because of new and harsh environments. Mimulus guttatus, a plant commonly seen on the west coast of the United States, is a prime example as it has rapidly evolved to soil contamination by copper due to mining in California in the last 150 years. There have been two hypotheses posed by researchers as to the genetic basis of how organisms have evolved so quickly which I set out to study: 1) There is a low frequency of tolerant genotypes in the ancestral population otherwise known as standing variation or 2) new mutations occurred once exposed to a new environment. In the past, researchers found it difficult to distinguish between the two because they lacked the technology we have today for DNA analysis. I used four different populations of M. guttatus from varying locations in order to address which hypothesis was valid. I conducted both survival assays of these populations and DNA analysis of known tolerant and non-tolerant lines using a copper oxidase gene. I found that there was at least some degree of tolerance in all populations in the survival assays, supporting the hypothesis of standing variation. I also found patterns within DNA analysis suggesting the copper oxidase gene would be useful for further study to verify the standing variation hypothesis. The results from this experiment helps in understanding rapid evolution not just in the context of soil contamination by metals but also ties back to why an alarming number of species are not able to adapt to our constantly changing world.

Hospital surfaces contamination with antineoplastic drugs: influence of cleaning procedures

Introduction: The raising frequency of cancer diseases is leading to a widespread application of antineoplastic drugs, thus increasing the probability of workplace surfaces contamination. Most of these drugs are classified by the International Agency for Research on Cancer as known or suspected human carcinogens. Skin absorption is the main route for antineoplastic drugs exposure in occupational settings, therefore cleaning protocols have paramount influence in surfaces contamination and, consequently, in exposure. The aim of this study was to assess surfaces contamination in a Portuguese chemotherapy unit before and during drug administration, in both preparation and administration facilities. Methods: Samples were collected by wipe-sampling from potentially contaminated surfaces selected by previous protocol observation. Samples were analyzed by HPLCDAD. Cyclophosphamide (CP), 5-fluorouracil (5FU), and paclitaxel (PTX) were used as surrogate markers for surfaces contamination for all cytotoxic drugs. Results: From the 34 samples collected before any preparation and administration activities, 41.2% were contaminated with 5-FU (4.0-84.7 ng/cm2) and 23.5% of the samples were contaminated with CP (19.8-139.6 μg/cm2). Only 2 samples presented contamination by PTX (5.9%) with a maximum value of 3.7 ng/cm2. Of the 37 samples collected during preparation and administration of antineoplastic drugs, 48.7% were contaminated with 5-FU (1.9-88.7 ng/cm2) and 24.3% with CP (12.0-93.9 μg/cm2). None of the samples showed contamination with PTX. Discussion: Data showed differences in contamination levels before and after the handling of antineoplastic drugs in preparation and in administration settings. These results point out the importance of cleaning procedures. This is well in accordance to previous studies that showed how the type of cleaning procedures and products used can be determinant for surfaces decontamination.