Science of the Total Environment 497–498 (2014) 679–687

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Environmental assessment of dredged sediment in the major LatinAmerican seaport (Santos, São Paulo — Brazil): An integrated approach

A. Cesar a,⁎, L.R.B. Lia b, C.D.S. Pereira a,b, A.R. Santos b, F.S. Cortez b, R.B. Choueri a, M.R. De Orte a, B.R.F. Rachid c

a UNIFESP, Universidade Federal de São Paulo, Depto de Ciências do Mar, Santos, SP, Brazilb UNISANTA, Universidade Santa Cecília, Laboratório de Ecotoxicologia, Santos, SP, Brazilc Fundação de Estudos e Pesquisas Aquáticas, São Paulo, SP, Brazil

H I G H L I G H T S

• Environmental assessment of a dredged sediment disposal area was performed.• The disposal area is altered with respect to sediment quality.• Sediment samples from the disposal area were toxic to sea-urchin and amphipods.• Application of multivariate analysis was useful to interpret the results.• This work may contribute to the new revisions of CONAMA Resolution No 454.

⁎ Corresponding author at: UNIFESP — UniversidadeCiências do Mar, Campus Baixada Santista, Av. Almiranteda Praia — Santos/SP— CEP 11030-400, Brazil. Tel.: +55

E-mail address: acesar@unifesp.br (A. Cesar).

http://dx.doi.org/10.1016/j.scitotenv.2014.08.0370048-9697/© 2014 Elsevier B.V. All rights reserved.

a b s t r a c t

a r t i c l e i n f o

Article history:Received 30 June 2014Received in revised form 8 August 2014Accepted 9 August 2014Available online xxxx

Editor: D. Barcelo

Keywords:Dredged materialSedimentContaminationToxicity testsEnvironmental assessmentMultivariate analyses

Thiswork offers an environmental assessment of a dredged sediment disposal area in Santos bay, situated on thecentral coast of the São Paulo State, Brazil. Sediment qualitywas evaluated through physicochemical analysis andtoxicity tests of sediments collected in the disposal site and adjacent area. The physicochemical characterizationof the sediments involved grain size distribution, concentrations of polycyclic aromatic hydrocarbons (PAHs),chlorinated and aromatic hydrocarbons, polychlorinated biphenyls (PCBs), pesticides, phthalates, metals and nu-trients. Acute and chronic toxicity tests were employed, using amphipods (Tiburonella viscana) and sea urchins(Lythechinus variegatus), respectively. Results revealed toxicity by all the methods applied here, suggestingthat the area of disposal of dredgedmaterial is significantly alteredwith respect to sediment quality and probablycapable of generating deleterious effects on the local biota. Aiming to elucidate the association between the dis-tinct environmental variables and the biological effects measured in laboratory, Factor Analysis was performed.Results revealed that despite most contaminant concentrations were found below the limits established byBrazilian legislation, biological effects were related to metals (chronic toxicity) and organic compounds (acutetoxicity). The application of multivariate analysis proved to be particularly useful to assess and interpret the re-sults in an integratedway, particularly due to the large number of parameters analyzed in environmental assess-ments, and should be applied in future studies.

© 2014 Elsevier B.V. All rights reserved.

1. Introduction

Seaports are one of the most important issues in the national andinternational sphere, particularly today, when the adoption of measuresthat promote development is considered crucial and represents a priorityissue for many countries' economic and social developments. The portof Santos plays a major role in Brazilian's national context, due to itssize and shipping capacity (Torres et al., 2009). This is the country'smain gateway for incoming and outgoing products.

Federal de São Paulo, Depto deSaldanha da Gama, 89 — Ponta13 3523 5094.

Besides its economic importance, the region is also environmentallyrelevant, since the Santos estuarine system is surrounded bymangroves, which make up 43% of the total area of marshlands in thestate of São Paulo (Lamparelli et al., 2001). This estuarine region wasimpacted over the years by the human activities of industry, harborand resident population in the surroundings and proximities of theestuary. As a result, huge quantities of pollutants were launched intothe estuary, causing high levels of contamination and toxicity of thesediments (Abessa, 2002; Buruaem et al., 2013; Cesar et al., 2006;Lamparelli et al., 2001; Rachid, 2002).

Maintenance of the port's activities and safe navigation in the Portof Santos requires constant dredging of both fairways and berthingdocks. This operation is a common practice around the world, given

680 A. Cesar et al. / Science of the Total Environment 497–498 (2014) 679–687

the characteristics of coastal water bodies, especially estuaries, whichare prone to natural sedimentation or to sedimentation accelerated byhuman activities. However, disposal of dredged material is regarded asone of the most important problems in coastal zone management(Perrodin et al., 2014).

Dredged material disposal pose risks to aquatic organisms at thedumping site as well as adjacent areas. Physical alterations and buryingof benthic habitats, increased water column turbidity, enhancement oforganic matter, nutrient and contaminant loads in sediments and theirdissolution are among the main sources of impacts to resident biota atdisposal sites (Bolam, 2014; Lions et al., 2010; Stronkhorst et al., 2003;Torres et al., 2009; Zimmerman et al., 2003).

Although in many countries (including Brazil) permissions tomarine disposal of sediments are only conceded after verification thatcontaminant levels are within specific quality standards, there is still arisk of harming resident biota at the disposal site through the mobiliza-tion of toxic compounds due to sediment oxidation during the dredgingand dumping processes and or physical–chemical differences of benthichabitat between the dredging and disposal site (e.g. redox potential,granulometry). In addition, Brazilian sediment quality guidelines(SQGs), given by the CONAMA 454/2012 (Brasil, 2012), are based oninternational standards, particularly the North American Effects RangeLow (ERL) and Effects Range Median (ERM) for marine and estuarinesediments (Long et al., 1995). Derived for the application in a differentenvironmental context, international guidelines may not suitablypredict ecotoxicological impacts in Brazilian aquatic environments(Choueri et al., 2009). A thorough environmental assessment is there-fore necessary at the disposal site in order to evaluate the potentialrisk of contaminants in the dredged material to the resident biota,especially when the dredged sediments come from contaminated sites.

Chemical analyses and toxicity tests have been recommended andutilized in projects of monitoring environmental quality at dredgedmaterial dumping sites (CEFAS, 2005; Chevrier and Topping, 1998;Stronkhorst et al., 2003; Torres et al., 2009). When these methods areapplied separately, the environmental quality description is oftenincomplete and inadequate (Blasco and Picó, 2009). However, when

Fig. 1. Location of the sampling points and adjacent area indic

applied in combination, they supply more realistic information aboutthe environmental conditions of a site (Cesar et al., 2007). Many studieshave integrated environmental data through multivariate analysis(Buruaem et al., 2013; Cesar et al., 2006, 2007; Choueri et al., 2010;DelValls et al., 2002; De Orte et al., 2014a,b; Paixão et al., 2011; Ribaet al., 2004a,b; Tlili et al., 2010; Zeng and Arnold, 2014), particularlyusing Principal Component Analysis (PCA). Such analysis allows adeeper understanding of the relationships among variables in thedataset with minimal loss of information, providing an objective proce-dure to achieve the final decision about the environmental risks ofsediments and dredged material.

In the present study, the environmental quality of the dredgedsediment disposal area and adjacent coastal zone located in SantosBay was assessed based on sediment physicochemical and ecotoxico-logical analysis. The data presented in this work is part of the firstcontinuous monitoring program from the Port of Santos. The resultsfrom the analysis were integrated using multivariate analysis tech-niques such as Factor Analysis (FA) and Principal Component Analysis(PCA) to identify the principal variables associated with the biologicaleffects that determine the environmental quality of the region understudy.

2. Materials and methods

2.1. Sampling

The information presented in this work is part of the processeSMA-13740/04, which belong to the licensing operations maintenancedredging in the port area of Santos, from the Department of Environ-ment State of São Paulo. These data are part of the monthly reportsmonitoring the disposal area for dredged material at the port of Santos,São Paulo — Brazil. The sediment dredged out from the Santos harborwas discharged in a square-shaped area delimited by the BrazilianNavy and the state environmental agency (CETESB), which is con-sidered as coastal area, with a water column of 22 m. The area (markedin Fig. 1 with a square) covers approximately 4 km2 and is situated

5 km

N

ating the quadrilateral disposal of the dredged material.

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between the latitudes 24°04′ S and 24°05′ S and the longitudes 46°15′W and 46°16′ W, about 20 km from the entrance to the port of Santos.

The study involved nine sediment sampling sites situated aroundSantos Bay, comprising the coastal zone of the municipalities of PraiaGrande, Santos, Guarujá and Bertioga (Fig. 1). The sediment sampleswere collected on November 16, 17 and 23, 2007.

The location of the sampling points was chosen according to thespatial gradient established by the monitoring plan for the disposalarea for dredged material at the harbor of Santos. Fig. 1 indicates thelocality of the sampling sites including the region of disposal of thedredged material, pointed out as a square. Site 9 was deemed a refer-ence site, e.g. not influenced by the dredgedmaterial disposal activities.

The sediment samples were collected with a stainless steel van Veengrab sampler with an area of 0.04m2, taken to the laboratory and storedin a cooler (4 °C) until analysis.

2.2. Physicochemical analysis of the sediments

The sediment samples were subjected to the following analyses:(a) granulometric distribution obtained and reported in line withCONAMA Resolution 454/12; (b) total organic carbon (TOC) deter-mined according to Verdardo et al. (1990); (c) total nitrogen (Kjeldahl)was quantified according to Standard Methods 4500-Norg B and 4500-NH3 C and G (APHA, 2005); (d) total phosphorus, determined by theascorbic acid method according to method 4500 PE (APHA, 2005);(e) metals (Cd, Pb, Cu, Cr, Mn, Ni and Zn) and As were determined byinductively coupled plasma (ICP) atomic emission spectroscopy, usingtheUS EPA 6010C (USEPA 2007a)method after digestion of the samplesby theUS EPA 3051Amethod (USEPA, 2007b).Mercury (Hg)was deter-mined in an atomic absorption spectrophotometer by the US EPA 7471method (USEPA, 1994); (f) polycyclic aromatic hydrocarbons (PAH)and phthalates, determined by gas chromatography coupled to massspectrometry (GC/MS) according to the US EPA 8270 method after ul-trasonic extraction by the US EPA 3550C method (USEPA, 2007c) andwashing in a silica gel column according to the US EPA 3630C method(USEPA, 1996a); (g) chlorinated hydrocarbons determined by gas chro-matography, with the chlorines determined using an electron capturedetector (USEPA 5021 CG/ECD method) (USEPA, 1996b); (h) organo-chlorine pesticides and phenolic compounds, determined by gas chro-matography according to the US EPA 8081B (USEPA, 2007d) and USEPA 8041Amethods (USEPA, 2007e); and (i) polychlorinated biphenyls(PCBs), determined by gas chromatography according to the US EPA8082Amethod (USEPA, 2007f), with the samples purified in an aluminacolumn (neutral pH) according to the US EPA 3611B method (US EPA,1996c).

Certified reference materials were used to check the quality of theanalytical procedures. For metals, it was utilized as reference, theMarine Sediment References Material for Trace Metals—1, NationalResearch Council (NRC), Certified Reference Material 277 BCR, andConseil National de Recherche Canada, 277 BCR. For organiccompounds, NRC-CNRC HS-1 was the reference material. Goodagreement with certified values was obtained, with recoveries rangingfrom 90% to 110%.

2.3. Toxicity tests

Three toxicity tests were carried out to represent different routes ofexposure: (i) whole sediment toxicity test, to assess the effects causedby thedirect contact of organismswith the sediments; (ii) sediment elu-triate toxicity test, which assesses the transference and bioavailability ofcontaminants from sediments to the water, after a resuspension pro-cess; and (iii) the sediment–water interface toxicity test, to evaluatethe toxicity of sediments due to arising fluxes of contaminants frompore water as well as desorbed contaminants.

2.3.1. Acute toxicity test with amphipodsThe method used in the toxicity test on whole sediment with the

burrowing amphipod Tiburonella viscana was adapted by Melo (1993)and used by Zaroni (2006). Three days before the beginning of theexperiment, specimens of T. viscana were collected on Engenho D'Águabeach, an uncontaminated site at the island of São Sebastião (SP). Theorganisms were acclimated to the laboratory by keeping them in atank containing clean water and a layer of approximately 1 cm ofsediment from the site where they were collected. The tank waskept under constant aeration and illumination at a temperature of25 ± 2 °C. After the acclimation period, the animals were selected forthe test.

Adult amphipods (except for ovated females and animals withaltered appearance or movement) were exposed to the sedimentsamples for 10 days. Sediments from the organism's sampling sitewere used as negative control. Prior to the experiment, the sedimentsamples were homogenized and four replicates of each were preparedin high density polyethylene jars containing 175 ml of sediment and750 ml of seawater. Ten amphipods were placed in each jar. All thesamples received constant aeration and illumination and controlledtemperature (25 ± 2 °C), in a climatized room.

Physicochemical parameters such as pH (Mettler ToledoMP 120 pHMeter), salinity (Atago Hand Refractometer S/Mill-E) and totalammonia (Ammonia Marine & Freshwater Test Lab-Red Sea) weremeasured in thewater from the jars at the beginning of the experiment.Furthermore, total ammonia was measured in the interstitial waterextracted by centrifugation, and non-ionized ammonia was estimatedfrom those measurements. Daily observations were made to check onthe presence of emerging organisms in each jar. At the end of the periodof exposure (10 days), sediments were sieved and the mortality ratewas recorded. The mean survival rate of the organisms of each samplewas compared with that observed in the reference sediment and inthe control sediment. The results were compared statistically by themethods of MSD (Minimum Significant Difference) and analysis ofvariance, followed by Dunnett's test to detect significant differencesand determine toxic samples (α = 0.05).

Concomitantly, a test was carried out with a reference substance,without the presence of sediment, to evaluate the sensitivity of theorganisms employed. The tested reference substance was potassiumdichromate (K2Cr2O7). After 48 h of exposure, a count was made ofthe number of live and dead organisms in each replicate, and theLC5048 h for potassium dichromate was calculated. The result obtainedin the test with this substance was analyzed by the TrimmedSpearman-Karber statistical method with an Abbott correction(Hamilton et al., 1977), to calculate the LC5048 h, i.e., the concentrationlethal to 50% of the organisms after 48 h, and its confidence interval.

2.3.2. Short-term chronic toxicity test with sea urchinsThe methodology used here was based on the Brazilian NBR 15350

standard (ABNT, 2012). Adult sea urchins of the species Lytechinusvariegatus (Lamarck, 1816) were collected by freediving at PalmasIsland situated on the Santos bay.

Elutriates were prepared using an aliquot of 150 g of sediment fromeach samplewhichwas homogenized for 30minwith 600ml of filtereddiluted seawater, using a Turbo-Flow 2c shaker (PoliControl) at aconstant speed of 105 rpm. After shaking, the samples were allowedto rest for 24 h, and then 10 ml of the supernatant was siphoned offand added to each replicate of the test, following the Brazilian ABNTNBR 15350 standard (ABNT, 2012).

For the sediment–water interface treatment, 2.0 ml of the sedimentwas added to each replicate, using a 5 ml syringe. A plankton net(45 μm) was then placed over the sediment and fixed with a plasticring, and 8.0 ml of diluted seawater was added, using an automaticpipette as described by Cesar et al. (2004). For both elutriate andsediment–water interface tests, negative control was done by using

682 A. Cesar et al. / Science of the Total Environment 497–498 (2014) 679–687

uncontaminated natural seawater. Four replicates were done for eachtest.

The following initial and final physicochemical measurements weretaken in the aqueous fraction during the toxicity testswith L. variegatus:temperature (using an INCOTERM glass thermometer), salinity (with aShibuya 145 refractometer), pH (with a Laborgraf B474 pH meter),dissolved oxygen (membrane electrode method, using an WTWOxi 3151 oxygen meter), and total ammonia following the standardmethod 4500-C (distillation and titration method) (APHA, 2005). Theconcentrations of non-ionizable ammonia were estimated from thesevalues.

The results of the testswith sediment–water interface and elutriatedsedimentwere subjected to a single analysis of variance (ANOVA), usingDunnett's test to compare the sampled sites against the reference site(Site 9) and Student's t-test for independent paired comparisons ofthe sites in relation to the control. Before their application, the datawere subjected to an evaluation of normality by the chi-square methodand homogeneity of variances by Bartlett's test. For any statistical test, ap = 0.05 was utilized.

2.4. Integration of the sediment's physicochemical andecotoxicological results

Aiming to obtain an integrated and more robust interpretation ofthe sediment quality descriptors, Factor Analysis (FA) method withextraction of the variable by Principal Component Analysis (PCA) wasperformed to elucidate the correlation between the distinct environ-mental variables and the biological effects measured in laboratory. Theoriginal set of data considered 3 biological parameters (% of amphipodmortality, % of embryolarval abnormality for sea urchins exposed toelutriate, and % of embryolarval abnormality for sea urchins exposedto sediment–water interface) and all the measured physicochemicalvariables that presented detectable values (granulometry, KjeldahlN, TOC, total P, total PAH, PCBs, phthalates, Aldrin, Pb, Cu, Cr, Mn,Ni, Zn). The FA was carried out with a matrix in which the variableswere standardized and factors were rotated using the varimaxnormalized procedure. In addition, the prevalence of each factor forevery sampling station in relation to all stations was also calculated(factor scores).

All the analyses were carried out using the MULTIVARIATEEXPLORATORY TECHNIQUES option followed by the Factors Analysisof the STATISTICA® software package (Stat Soft, Inc. 2013; version12), from which the principal factors were extracted and the eigen-values above 1.0 taken into account (Kaiser's criterion). A componentloading cutoff of 0.45 was used in selecting variables for inclusion infactors. Tabachinic and Fidell (1996) suggested that a cutoff of atleast 0.32 be used and that component loading of greater than 0.45 beconsidered fair or better.

Table 1Percentage of fine grains and concentrations of nutrients, PAHs, PCBs, phthalates, and heavy m

Sites Va

Fines(%)

Kjeldahl N(mg/kg)

TOC(%)

Total P(mg/kg)

Total PAHs(μg/kg)

PCBs(μg/kg)

Phthalates(μg/kg)

1 8.2 47.2 0.28 12.8 0 0 02 8.2 88.3 0.3 17.2 0.5 0 03 19.5 71.2 1.36 8 576.5 182 524 57.7 270 3.64 25.1 118.8 0 685 60.2 392 3.16 38.6 57 60.9 06 33.2 223 1.11 25.9 12.5 14.9 07 28.9 55.8 0.96 19.3 59.3 23.4 08 26.1 39.3 0.8 15.6 48.8 26.3 09 12.5 56 0.71 8.7 14.2 10.4 0

3. Results and discussion

3.1. Physicochemical analyses and comparison against national andinternational sediment quality guidelines

Table 1 lists the results of the percentage of fine grains in thesediments, the nutrient concentrations (N Kjeldahl, TOC and total P),the concentrations of organic compounds (total PAHs, PCBs, phthalatesand Aldrin), and the concentrations of metals detected at each samplingsite. The granulometric analysis revealed a predominance of fine grains(silt + clay) at sites 4 and 5, adjacent to the dredged sediment disposalsite, and a predominance of sand at the remaining sampling sites.

The following nutrients were detected at all the sampling sites: 39.3to 392.0 mg kg−1 of total Kjeldahl nitrogen; 0.28 to 3.64% of TOC; and8.0 to 38.6 mg kg−1 of total phosphorus, all of which were lower thanthe warning level established by the Brazilian legislation regardingdredged material characterization and management (CONAMA Resolu-tion 454/12, Supplementary materials — SM, Table S1) (Brasil, 2012).

The metals Pb, Cr, Mn, Ni and Zn were detected at all sampling siteswhereas Cd, Hg and As were not detected in any sample. The metalconcentrations in this study showed values below level 1 of CONAMA454/12. They were also low compared to national action levels fordredged material applied in different countries such as Canada,Australia and European countries (ANZECC/ARMCANZ, 2000; CCME,2002; OSPAR, 2008— SM, Tables S2–S4) and commonly used guidelinesbased on biological effects, i.e. Apparent Effects Threshold-Low/Apparent Effects Threshold-High (AET-L/AET-H) (USEPA, 1991), EffectsRange-Low/Effects Range-Median/(ERL/ERM) (Long et al., 1995),Threshold Effects Level/Probable Effects Level (TEL/PEL) (Macdonaldet al., 1996).

However, the commonly used sediment quality guidelines (SQGs),may not suitably address specificities of different environmental charac-teristics. Although these values were derived from ample databases ofcontaminant levels and their related biological effects, disparities arefound between the classical SQGs and those derived specifically for aparticular area. Choueri et al. (2009) developed SQGs specifically forthe Santos estuarine system, based on available data from previousstudies. Likewise for other SQGs, each individual SQG derived for Santosestuarine system provides a lower limit and an upper limit. It is impor-tant to consider that, for some contaminants, the specific SQGs areduplicated since two different data setswere used to derive such values.Thus, for an easier comparison of the values found in the present study,the duplicated SQGs were combined into one and four limits wereestablished, ranging from the 1st level (below which sediments areconsidered ‘not polluted’ since both SQGs define as ‘not polluted’) tothe 4th level (above which sediments are considered ‘polluted’ sinceboth SQGs define as ‘polluted’ sediments). Within this range, sedimentsshowing concentrations falling in between the 1st and 2nd levels areclassified as ‘low to moderately polluted’ (i.e. one SQG classifies as low

etals in the nine sampling sites.

riables

Aldrin(μg/kg)

Pb(mg/kg)

Cu(mg/kg)

Cr(mg/kg)

Mn(mg/kg)

Ni(mg/kg)

Zn(mg/kg)

0.14 7.8 0 9.9 135 2.5 22.30.07 6.1 0 10 136 2.9 181.14 12 3.2 18 392 6.9 360.3 23 11 34 713 13 620.36 23 10 33 652 13 590.09 13 3 21 387 7.8 380.16 8 0 16 271 5.6 290.08 9.3 0.5 15 273 5.3 36.70.05 6.2 0 12 178 3.8 26.8

Table 2Results of the toxicity tests.

Sites Variables

% Amphipodmortality

% Embryolarvalanomaly elutriate

% Embryolarvalanomaly interface

1 27.5 21 42.62 22.5 36.4 40.23 70 86.6 97.44 57.5 99.8 98.65 30 55.8 93.26 57.5 48.6 93.87 27.5 50 61.88 25 53.4 589 35 23 35.6

Table 3Estimated loadings for the nine sediment sampling sites.

Variables Factor loadings (Varimax normalized)(FQ-toxicity-17)Extraction: principal components(Bold - marked loadings are N .450000)

Factor 1 Factor 2

N 0.93883 −0.056861TOC 0.94572 0.236726P 0.87102 −0.319477% Fine grains 0.97973 0.056767Mortality — amphipod 0.25942 0.806814Elutriate — sea urchin 0.57742 0.713901Interface — sea urchin 0.73271 0.578755PAHs −0.06576 0.983330PCBs −0.05009 0.873479Phthalates 0.36332 0.767636Aldrin 0.05753 0.948254Pb 0.96613 0.212885Cu 0.94985 0.230815Cr 0.97813 0.199518Mn 0.94829 0.311743Ni 0.96797 0.237055Zn 0.94970 0.226546

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polluted and the other as moderately polluted), between 2nd and3rd levels are ‘moderately polluted’ (range in which both SQGs classifysediments as moderately polluted), and between 3rd to 4th areconsidered as ‘moderately to highly polluted’ (i.e. one SQG classifies asmoderately polluted and the other as highly polluted). Following suchclassification, sites 4 and 5 are in the worst condition regardingmetal contamination, since these sites are highly polluted by Pb andmoderately to highly polluted by Ni; also, Zn is at moderate level insite 4 and low to moderate in site 5. Site 3 and site 6 are moderatelyto highly polluted by Ni as well, and low to moderately polluted by Pb.Site 6 is also low to moderately polluted by Zn. Lastly, sites 7 and 8 arelow to moderately polluted by Ni.

Sites 2 to 9 showed total PAHs in the range of 0.5 to 576.5 μg kg−1.The highest value was found in site 3. Notar et al. (2001) established acriterion to evaluate the levels of pollution of total PAHs in marinesediments and suggested that sediments with concentrations ofΣPAHs b 250 μg kg−1 are considered little polluted, while concentra-tions of 250 b ΣPAHs b 500 μg kg−1 correspond to moderately pollutedsediments and sediments with concentrations of ΣPAHs N 500 μg kg−1,which is the case of site 3 in the present study, are considered highlypolluted. Comparing to the SQGs specifically derived for Santos estua-rine system (Choueri et al., 2009), total PAHs levels observed in thisstudy do not represent high ecotoxicological risk in any studied site;only site 3 is moderately polluted by total PAHs. However, a differentscenario is depicted when PAHs are analyzed individually. In this case,the concentration of chrysene in site 3 (104.4 μg/kg) (SM, Tables S1)is higher than the value established by the Centre for Environment, Fish-eries and Aquaculture Science (CEFAS) for characterizing dredged ma-terial in the UK (OSPAR, 2008). Site 3 is also contaminated bydibenzo(a,h)anthracene (37.6 μg/kg) (SM, Tables S1) at a level that vio-lates not only CEFAS level for this compound, but also the TEL, utilizedby the Canadian Council of Ministers of the Environment as sedimentquality guideline (CCME, 2002). Nevertheless, these values are belowlevel 1 of CONAMA 454/12.

Among the phthalate compounds analyzed, only bis(2-ethylhexyl)phthalate was detected in the samples from sites 3 and 4, in concentra-tions of 52.0 and 68.0 μg kg−1, respectively (SM, Tables S1). These con-centrations fall below the TEL (threshold effect level) of 182 μg kg−1

developed for Florida coastal waters (MacDonald, 1994), whileCONAMA Resolution 454/12 does not establish classification limits forphthalates (SM, Tables S1).

Among chlorinated hydrocarbons, only heptachlor epoxidewas detected in sites 3–5. The level of this compound in site3 (3.2 μg kg−1) is higher than PEL (2.74 μg kg−1), although this guide-line value, provisionally applied for characterizing Canadian marinesediments (CCME, 2002), was actually developed for freshwatersediments (SM, Tables S1). Brazilian legislation does not establish actionlevels for this compound. Heptachlor epoxide is the degradationproduct of the synthetic organochlorine pesticide heptachlor. Eventhough the use of heptachlor was prohibited (or severely restricted)in several countries, including Brazil (Brasil, 1985), this compoundand its degradation products are still found in different environmentalmatrices of Brazilian aquatic systems (Yogui et al., 2003; Corbi et al.,2006) and estuarine systems worldwide (Alvarez Piñeiro et al., 1995;Lanfranchi et al., 2006).

Of the chlorinated pesticide group, Aldrin was found at the ninesampling sites in concentrations of 0.05 μg kg−1 to 0.36 μg kg−1.Endosulfan sulfate, in turn, was detected only in sites 2 (3.2 μg kg−1)and 3 (0.7 μg kg−1), while Mirex was found at most of the samplingsites, except for 3, 7 and 8, in concentrations varying from 0.16 μg kg−1

to 0.63 μg kg−1. No official guidelines of sediment/dredged materialquality exist for the above mentioned compounds at national level.According to Persaud et al. (1993), the minimum concentrations ofAldrin and Mirex that pose biological risks are 2 and 7 μg kg−1, respec-tively, which exceed the values detected in this study. Also in the chlo-rinated pesticide group, it was detected 0.1 μg kg−1 of delta-BHC in site

4, 0.2 μg kg−1 of gamma-Chlordane in site 7, 1.0 μg kg−1 of DDE in sites4 and 5, and 1.0 μg kg−1 of DDT in site 5. All the results fell below theinternational quality standards for sediments/dredged material quality,as well as level 1 of CONAMA Resolution 454/12. BTEX and phenoliccompounds were not detected in any sample in the study area (SM,Tables S1).

In the case of the PCBs, polychlorinated biphenyls were found at sixsampling sites, three of which (5, 7, and 8) showed levels between level1 and level 2 (i.e. the lower and upper action levels) of CONAMAResolution 454/12, which is 22.7 and 180 μg kg−1, respectively, andtherefore they pose some risk to the aquatic biota. PCB levels at thesesites also exceed TEL (CCME, 2002), ERM (Long et al., 1995) and theAustralian guideline for dredged material characterization (ANZECC/ARMCANZ, 2000). A higher value was found at site 3 (182 μg kg−1),the disposal site of dredged sediment, exceeding level 2 of CONAMAResolution that represents a high ecological risk.

3.2. Toxicity tests

Table 2 presents the results of amphipodmortality and embryolarvaldevelopment of sea urchins. Using the statistical package Toxstat, wefound that the result of the test was normal and homogeneous. UsingANOVA followed by Dunnett's test, and considering site 9 as reference,the toxicity test with whole sediment showed an effect on the survivalof the test organism only in the sample from site 3 for the amphipodT. viscana. However, the mean survival rate of the reference sample

684 A. Cesar et al. / Science of the Total Environment 497–498 (2014) 679–687

(site 9)was low; therefore this samplewas also included in the statisticalanalysis. According to this new analysis, the sites 3, 4 and 6 presentedsignificant effects on the survival of the test organisms when comparedto the control (sediment from Engenho D'Água beach), thus, these siteswere considered toxic to the amphipod T. viscana. The values of non-ionized ammonia, NH3, estimated at the beginning and end of thetests were lower than those suggested by Sáfadi et al. (2004), whostated that the sensitivity of the amphipod T. viscana to ammoniumchloride was 0.3 mg kg−1. The 48-h LC50 estimated for the K2Cr2O7

was 17.47 mg/L. This result indicates that the amphipods used inthe test showed a sensitivity range within the expected limits for thespecies (Abessa and Sousa, 2003).

The short-term chronic toxicity tests with elutriate sedimentpresented a significantly deleterious effect on the embryolarvaldevelopment of L. variegatus in the samples from sites 3 to 8. The results

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SITE 4 SIT

SITE 7 SIT

Fig. 2. Scores of each factor at

of the short-term chronic toxicity test with sediment–water interfacealso showed an effect on the embryolarval development ofL. variegatus at sites 3–8. Site 9 was used as reference in both methods.With regard to the physicochemical analyses carried out during thetest with samples of elutriate and sediment–water interface, theinitial values of temperature, dissolved oxygen, pH, salinity, totalammonia and non-ionized ammonia fell within the threshold range ofthe species (ABNT NBR 15350; Prósperi, 2002). The results of thecorrelations drawn among the various methods adopted here toevaluate the toxicity were consistent with regard to the sampling sitesthat presented adverse biological effects. Sites 3, 4 and 6 revealedtoxicity by all the methods applied, suggesting that the area of disposalof dredged material is significantly altered with respect to sedimentquality and probably capable of generating deleterious effects on thelocal biota.

FA

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E 5 SITE 6

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the nine sampling sites.

685A. Cesar et al. / Science of the Total Environment 497–498 (2014) 679–687

3.3. Integration of the sediment's physicochemical andecotoxicological results

The results from the Factor Analysis are shown in Table 3. Two newfactors were obtained from the original values. These factors explainedtogether 90.15% of the variance in the original set of data. The first factorwas responsible for 65.86% of the variance of the data, while the secondfactor accounted for 24.18% (SM, Tables S5).

As showed in Table 3, the predominant factor (Factor 1) groupstogether the granulometry (% fine grains), nutrient concentrations(Kjeldahl N, TOC, total P) and the metals (Pb, Cu, Cr, Mn, Ni, Zn) associ-ated with the effect on the embryolarval development of the sea urchin(elutriate and sediment–water interface). Sulfur speciesweremeasuredin previous studies (Torres, 2007 and Torres et al, 2009) and showedlower concentrations in the disposal site and adjacent area, whichindicated that contaminants were not bioavailable. Fig. 2 representsthe estimated scores of each factor to each sampling site. It is observedthat factor 1 is representative for sites 4–6.

This result suggests that some toxicity was found in associationwithmetal contaminants in the sampling sites 4–6. As discussed afore,although the levels of metals in the sampling stations were not exceed-ing CONAMA 454/12 or classical SQGs, according to the classificationgiven by SQGs specifically derived for Santos Estuarine System, theconcentrations of some metals may be considered harmful to thebiota, mostly in sites 4–6. Therefore, the results of the PCA are in betteragreement with the specific SQGs compared with the classical SQGs. Aqualitative evaluation of the PCA results shows that the metals whichare more likely to be causing the observed toxicity in sites 4–6 are Pb,Ni, and Zn. The association between the other metals (Cu, Cr, and Mn)and toxicity, showed by the PCA, may be merely a mathematicalcovariance among variables.

Factor 2, in turn, associates the concentrations of total PAHs, PCBs,Aldrin and phthalates with the biological effects of amphipod mortalityand sea urchin embryolarval development (elutriate and sediment–water interface). The scores of sites 3 and 4 are positive for factor 2,which means that the associations revealed by factor 2 are relevant forthese sampling sites (Fig. 2). This suggests an association betweentoxicity and levels of organic contaminants (PAHs, PCBs, Aldrin andphthalates). The environmental situation is clearer for site 3, wherethe toxicity is probably caused by PCBs, since the level of such contam-inant exceeds the highest level of national and international SQGs, andPAHs, given that this group of contaminants exceeds the SQG-minimum derived specifically for Santos Estuarine System. In site 4,the role of the associated contaminants (PAHs, PCBs, phthalates, Aldrin)

6

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8 71

92

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Fig. 3. Scores of the nine collection sites distribu

in factor 2 on the toxicity is less clear. Levels of PAHs are quite low andPCBs were not found in this site; also, although phthalates and Aldrinlevels are between the highest concentrations among sampling sitesin this study, they arewell below the existing SQGs (derived by interna-tional agencies). Therefore, it is reasonable to consider two options:either the existing SQGs for phthalates and Aldrin are not suitable forapplication in Santos Estuarine System, as it occurs for other contami-nants, or the cause of the observed toxicity is exclusively the metalsdiscussed above, and the estimated factor score showing a prevalenceof factor 2 to site 4 is a mathematical artifact.

Fig. 3 depicts the scores from the nine collection sites, distributed intwo-dimensional space according to their association with the twoprincipal components. This analysis indicates the formation of threedistinct groups. One group is composed of sites 1, 2, 7, 8 and 9 withnegative scores for both factors. Those sites are characterized by agreater distance from the dredge material disposal area. According tothe analysis performed in this study, these sites appear to hold a betterenvironmental condition than the other sites and are probably notinfluenced by the disposal area. The second group comprises sites 4, 5and 6 with positive scores for factor 1. This group of sites is includedwithin the zone of influence of the discharged dredged material, beingcontaminated,mostly bymetals, while this contaminationwas correlat-ed with toxic responses to marine organisms. Therefore, sediment fromthese sites presents significant ecological alterations. The last group isrepresented by site 3, the dredged material disposal area, which isseparated from the others and scored positive for factor 2. This sitealso indicates alterations in sediment quality, which could be able togenerate ecological disturbances. This was demonstrated by toxicityresponses associated with high concentrations of PBCs and PAHs, andpossibly by Aldrin and phthalates.

4. Conclusions

In the present study, the assessment of the quality of sediments froma dredged material disposal area showed that despite most contami-nants concentrations were found below the limits established byBrazilian legislation, biological effects were detected and related tometals (chronic toxicity) and organic compounds (acute toxicity).Therefore, it can be concluded that the area utilized to dispose dredgedmaterial from the Santos harbor (site 3) and the immediately adjacentarea (Sites 4, 5 and 6) presents significant alterations in terms ofsediment quality, which could be able to generate ecological distur-bances. According to these results we suggest that using chemicalconcentration as the first step in the management procedures might

4

51,0 1,5 2,0

ted in two-dimensional space (PC1 × PC2).

686 A. Cesar et al. / Science of the Total Environment 497–498 (2014) 679–687

not be environmentally protective, while the use of site-specific SQG ishighly recommended. Furthermore, the application of multivariateanalysis proved to be particularly useful to assess and interpret theresults in an integrated way, particularly due to the large number ofparameters analyzed in environmental assessments, and should beapplied in future studies.

Acknowledgments

We are grateful to the anonymous referee for his useful commentsand constructive suggestions. Cesar and Pereira thanks to CNPq(Conselho Nacional de Desenvolvimento Científico e Tecnológico, MEC —

Brazil) for the research productivity scholarships (PQ#305869/2013-2and PQ#307074/2013-7, respectively). De Orte thanks to CAPES(Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, MEC —

Brazil) for the postdoctoral scholarship. The authors declare that thisstudy was conducted in accordance with the national and institutionalguidelines for the protection of human subjects and animal welfare.

Appendix A. Supplementary data

Supplementary data to this article can be found online at http://dx.doi.org/10.1016/j.scitotenv.2014.08.037.

References

Abessa DMS. Avaliação da Qualidade de Sedimentos do Sistema Estuarino de Santos, SP,Brasil [Tese de doutorado] São Paulo: Instituto Oceanográfico da Universidade deSão Paulo; 2002 [124 pp.].

Abessa DMS, Sousa ECPM. Sensitivity of the amphipod Tiburonella viscana to K2Cr2O7. BrazArch Biol Technol 2003;46(1):3–55.

ABNT NBR 15350 Associação Brasileira de Normas Técnicas. Ecotoxicologia aquática –

Toxicidade crônica de curta duração – Método de ensaio com ouriço-do-mar(Echinodermata; Echinoidea); 2012 [São Paulo].

Alvarez Piñeiro ME, Lozano JS, Yusty MAL. Organochlorine compounds in mussels of theestuarine bays of Galicia (north-west Spain). Mar Pollut Bull 1995;30:484–7.

ANZECC/ARMCANZ. Australian and New Zealand Guidelines for Fresh and Marine WaterQuality; 2000.

APHA. Standard methods for the examination of water and wastewater. 21st ed. AWWA,WEF; 2005 [1368 pp.].

Blasco C, Picó Y. Prospects for combining chemical and biological methods for integratedenvironmental assessment. Trends Anal Chem 2009;28(6):745–57.

Bolam SG. Macrofaunal recovery following the intertidal recharge of dredged material: acomparison of structural and functional approaches. Mar Environ Res 2014;97:15–29.

Brasil. Resolução CONAMA No 329, de 02 de setembro de 1985. Dispoõe sobre a proibição,em todo território nacional, a comercialização, o uso e a distribuição dos produtosagrotóxicos organoclorados, destinado a agropecuária dentre outros. Brasília, DF:Conselho Nacional de Meio Ambiente (CONAMA); 1995.

Brasil. Resolução CONAMA No 454, de 01 de novembro de 2012. Estabelece as diretrizesgerais e os procedimentos referenciais para o gerenciamento do material a serdragado em águas sob jurisdição nacional. Brasília, DF: Conselho Nacional de MeioAmbiente (CONAMA); 2012.

Buruaem LM, Castro IB, Hortellani MA, Taniguchi S, Fillman G, Sasaki ST, et al. Integratedquality assessment of sediments from harbor areas in Santos-São Vicente EstuarineSystem. Estuar Coast Shelf Sci 2013;130:179–89.

Canadian Council of Ministers of the Environment (CCME). Canadian Sediment QualityGuidelines for the Protection of Aquatic Life, Canadian Environmental QualityGuidelines, Summary Tables. Available in: http://www.ccme.ca/publications, 2002.

CEFAS. Environmental impacts resulting from disposal of dredged material at the RameHead disposal site. S.W. England: An analysis of existing data and implications forenvironmental management. Lowestoft: A CEFAS Multi-disciplinary Project Team.CEFAS; 2005. p. 120.

Cesar A, Marín A, Marin-Guirao L, Vita R. Amphipod and sea urchin testes to assess thetoxicity of Mediterranean sediments: the case of Portmán Bay. Sci Mar 2004;68(1):205–13.

Cesar A, Pereira CDS, Santos AR, Abessa DMS, FernándezN, Choueri RB, et al. Ecotoxicologicalassessment of sediments from the Santos and São Vicente estuarine system— Brazil.Braz J Oceanogr 2006;54(1):55–63.

Cesar A, Choueri RB, Riba I, Moralles-Caselles C, Pereira CDS, Santos AR, et al. Comparativesediment quality assessment in different littoral ecosystems from Spain (Gulf ofCadiz) and Brazil (Santos and São Vicente estuarine system). Environ Int 2007;33:429–35.

Chevrier A, Topping PA. National Guidelines for Monitoring Dredged and ExcavatedMaterial at Ocean Disposal Sites. Environment Canada, Marine Environment Division;1998 [27 pp.].

Choueri RB, Cesar A, Abessa DMS, Torres RJ, Morais RD, Riba I, et al. Development of site-specific sediment quality guidelines for North and South Atlantic littoral zones:

comparison against national and international sediment quality benchmarks.J Hazard Mater 2009;170(1):320–31.

Choueri RB, Cesar A, Abessa DMS, Torres RJ, Riba I, Pereira CDS, et al. Development of site-specific sediment quality guidelines for North and South Atlantic litoral zones: com-parison against national and international sediment quality benchmarks. Ecotoxicol-ogy 2010;19:678–96.

Corbi JJ, Strixino ST, Santos A, Del Grand M. Environmental diagnostic of metals andorganochlorinated compounds in streams near sugar cane plantations activity (SãoPaulo State, Brazil). Quim Nova 2006;29(1):61–5.

De Orte MR, Lombardi AT, Sarmiento AM, Basallote MD, Rodriguez-Romero A, Riba I, et al.Metal mobility and toxicity to microalgae associated with acidification of sediments:CO2 and acid comparison. Mar Environ Res 2014a;96:136–44.

De Orte MR, Sarmiento AM, Basallote MD, Rodríguez-Romero A, Riba I, DelValls A. Effectson themobility of metals from acidification caused by possible CO2 leakage from sub-seabed geological formations. Sci Total Environ 2014b;470–471:356–63.

DelValls TA, Forja JM, Gómez-Parra A. Seasonality of contamination, toxicity, and qualityvalues in sediments from littoral ecosystems in the Gulf of Cádiz (SW Spain).Chemosphere 2002;46:1033–43.

Hamilton MA, Russo RC, Thurston RV. Trimmed Speraman–Karber method for estimatingmedian lethal concentration in toxicity bioassays. Environ Sci Technol 1977;11(7):714–9.

Lamparelli MC, Costa MP, Prósperi VA, Bevilacqua JE, Araújo RPA, Eysink GGJ, et al.Sistema estuarino de Santos e São Vicente. São Paulo: Relatório Técnico CETESB;2001 [183 pp.].

Lanfranchi AL, MenoneML, Miglioranza KS, Janiot LJ, Aizpun JE, Moreno VJ. Striped weak-fish (Cynoscion guatucupa): a biomonitor of organochlorine pesticides in estuarineand near-coastal zones. Mar Pollut Bull 2006;52:74–80.

Lions J, Guérin V, Bataillard P, Lee J, Laboudigue A. Metal availability in a highlycontaminated, dredged-sediment disposal site: field measurements and geochemicalmodeling. Environ Pollut 2010;158:2857–64.

Long ER, MacDonald DD, Smith SL, Calder FD. Incidence of adverse biologicaleffects withinranges of chemical concentrations in marine and estuarine sediments. EnvironManag 1995;19:81–97.

MacDonald DD. Approach to the assessment of sediment quality in Florida coastal waters.Development and Evaluation of Sediment Quality Assessment Guidelines, Florida, vol.1; 1994 [126 pp.].

MacDonald DD, Carr RS, Calder FD, Long ER, Ingersoll CG. Development and evaluation ofsediment quality guidelines for Florida coastal waters. Ecotoxicology 1996;5:253–78.

Melo SLR. Testes de toxicidade com sedimentos marinhos: adequação de metodologiapara anfípodo escavador Tiburonella viscana [Dissertação de mestrado] São Carlos:Universidade de São Paulo, Escola de Engenharia de São Carlos; 1993 [172 pp.].

Notar M, Leskovek H, Faganeli J. Composition, distribution and sources of polycyclicaromatic hydrocarbons sediments of the Gulf of Trieste, Northern Adriatic Sea. MarPollut Bull 2001;42:36–44.

OSPAR. Assessment of the environmental impact of dredging for navigational purposes.OSPAR Commission, Publication nr 366/2008; 2008 [17 pp.].

Paixão JF, Oliveira OMC, Dominguez JML, Almeida ES, Carvalho GC, Magalhães WF.Integrated assessment of mangrove sediments in the Camamu Bay (Bahia, Brazil).Ecotoxicol Environ Saf 2011;74:403–15.

Perrodin Y, Donguy G, Emmanuel E, Winiarski T. Health risk assessment linked to fillingcoastal quarries with treated dredged seaport sediments. Sci Total Environ 2014;485–486:387–95.

Persaud D, Jaagumagi R, Hayton A. Guidelines for the protections and managementof aquatics sediments quality in Ontário. Toronto, Canada: Ontario Ministry ofEnvironment and Energy; 1993 [23 pp.].

Prósperi VA. Comparação de métodos ecotoxicológicos na avaliação de sedimentosmarinhos e estuarinos [Tese de Doutorado] São Carlos: Escola de Engenharia de SãoCarlos - USP; 2002.

Rachid BRF. Avaliação ecotoxicológica dos efluentes domésticos lançados pelos sistemasde disposição oceânica da Baixada Santista [Tese de Doutorado apresentada ao] SãoPaulo: Instituto Oceanográfico da Universidade de São Paulo; 2002 [286 pp.].

Riba I, Casado-Martínez C, Forja JM, Delvalls TA. Sediment quality in the Atlantic coast ofSpain. Environ Toxicol Chem 2004a;85:141–56.

Riba I, Forja JM, Gómez-Parra A, Delvalls TA. Sediment quality in littoral regions of theGulf of Cádiz: a triad approach to address the influence of mining activities. EnvironPollut 2004b;132:341–53.

Sáfadi RS, Melo SLR, Fontes AFÃ, Simões AP, Gelb EC, Costa FA, et al. Sensibilidade doanfípodo marinho Tiburonella viscana ao cloreto de amônia. Congresso Brasileiro deEcotoxicologia, 8. ; 2004. p. 48. [Florianópolis, B5-22].

Stronkhorst J, Ariese F, van Hattum B, Postma JF, de Kluijver M, Den Besten PJ, et al.Environmental impact and recovery at two dumping sites for dredged material inthe North Sea. Environ Pollut 2003;124:17–31.

Tabachinic BG, Fidell LS. Using multivariate statistics. New York, NY, USA: Harper Collins,College Publishers; 1996.

Tlili S, Jebali J, Banni M, Haouas Z, Mlayah A, Helal AN, et al. Multimarker approachanalysis in common carp Cyprinus carpio sampled from three freshwater sites.Environ Monit Assess 2010;168:285–98.

Torres RJ. Efeitos dadragagemsobre a qualidadedos sedimentos do canal do Porto de Santos(SP) [Tese de doutorado] Universidade Federal de São Carlos UFSCar; 2007 [159 pp.].

Torres RJ, Abessa DMS, Santos FC, Maranho LA, Davanso MB, Nascimento MRL, et al.Effects of dredging operations on sediment quality contaminantmobilization in dredgedsediments from the Port of Santos, SP, Brazil. J Soils Sediments 2009;9:420–32.

USEPA. (United States Environmental Protection Agency)/USACE (United States ArmyCorps of Engineers). Evaluation of dredged material proposed for ocean disposal —testing manual, EPA-503-8-91/001. Washington, D.C: Environmental ProtectionAgency Office of Marine and Estuarine Protection; 1991.

687A. Cesar et al. / Science of the Total Environment 497–498 (2014) 679–687

USEPA. Mercury in solid or semisolid waste (manual cold-vapor technique). SW-846,Method 7471. United States Environmental Protection Agency; 1994.

USEPA. Silica gel clean-up. Method 3630c Revision 3. United States EnvironmentalProtection Agency; 1996a [12 pp.].

USEPA. Volatile organic compounds in soils and other matrices using equilibriumheadspace analysis. SW-846, Method 5021. United States Environmental ProtectionAgency; 1996b [13 pp.].

US EPA. Alumina column cleanup and separation of petroleum wastes. Method 3611B,Revision 2. United States Environmental Protection Agency; 1996c [7 pp.].

USEPA. Determination of inorganic analytes by inductively coupled plasma-atomic emis-sion spectrometry. SW-846, Method 6010C, Revision 3. United States EnvironmentalProtection Agency; 2007a [34 pp.].

USEPA. Microwave assisted acid digestion of sediments, sludges, soils, and oils. SW-846,Method 3051A, Revision 1. United States Environmental Protection Agency; 2007b[30 pp.].

USEPA. Ultrasonic extraction. Method 3550c Revision 3. United States EnvironmentalProtection Agency; 2007c [17 pp.].

USEPA. Organochlorine pesticides by gas chromatography. Method 8081B, Revision 2.United tates Environmental Protection Agency; 2007d [57 pp.].

USEPA. Phenols by gas chromatography by gas chromatography. Method 8041A, Revision1. States Environmental Protection Agency; 2007e [30 pp.].

USEPA. olychlorinated Biphenyls (PCBs) by gas chromatography.Method 8082A, Revision1. States Environmental Protection Agency; 2007f [56 pp.].

Verdardo DJ, Forelich PN, Mc Intyre A. Determination of organic carbon and nitrogen inmarine sediments using the Carlo Erba NA-1500 analyzer. Deep-Sea Res 1990;37:157–65.

Yogui GT, Santos MCO, Montone RC. Chlorinated pesticides and polychlorinatedbiphenyls in marine tucuxi dolphins (Sotalia fluviatilis) from the Cananéia estuary,Southeastern Brazil. Sci Total Environ 2003;312:67–78.

Zaroni LP. Avaliação da qualidade dos sedimentos marinhos e estuarinos noMunicípio deBertioga-SP [Tese de Doutorado] Universidade de São Paulo. Instituto Oceanográfico;2006 [193 pp.].

Zeng T, Arnold WA. Clustering chlorine reactivity of haloacetic acid precursors in InlandLakes. Environ Sci Technol 2014;48:139–48.

Zimmerman LE, Jutte PC, Van Dolah RF. An environmental assessment of the CharlestonOcean Dredged Material Disposal Site and surrounding areas after partial completionof the Charleston Harbor Deepening Project. Mar Pollut Bull 2003;46:1408–19.