Marine Pollution Bulletin 162 (2021) 111907

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Marine Pollution Bulletin

journal homepage: www.elsevier.com/locate/marpolbul

Assessment of the causes of Hg bioaccumulation in the fish of a

Mediterranean lagoon subject to environmental management interventions
M. Lenzi a, *, E. Franchi b, M. Leporatti-Persiano c, A. D’Agostino d, P. Gennaro e, L. Marsili b
a
Lagoon Ecology and Aquaculture Laboratory (LEALab - WWF Oasi), Strada Provinciale Giannella 154, 58015 Orbetello, Italy
b
Department of Physical Sciences, Earth and Environment, University of Siena, Via Mattioli 4, Siena, Italy
c
Orbetello Pesca Lagunare, Via Leopardi 9, 58015 Orbetello, Italy
d
Department of Business and Quantitative Studies, University of Naples Parthenope, Italy
e
ISPRA, Italian Institute for Environmental Protection and Research, Leghorn, Italy

A R T I C L E I N F O A B S T R A C T

Keywords: In a Mediterranean lagoon characterized by high levels of Hg in the sediment, Hg content in commercial fish
Mercury species was determined, and, in order to establish Hg concentration in the water column, Diffusive Gradient in
Heavy metal Thin films technique (DGT) was used. The results allowed to state (1) the disturbance of the surface sediment and
Fish contamination
its resuspension in the water column did not cause detectable increase in Hg releases, (2) the East basin acted
DGT
more than the West one in contamination of fish species, (3) the small-sized sea-breams (<400 g) were more
Lagoon
Eutrophication contaminated than larger sizes ones (>400 g). We concluded: (1) fish contamination may also depend on direct
contact with releases of the contaminant and small, gregarious and less sedentary subjects are more likely to
incur releases of Hg; (2) Hg releases can be greater in areas with a relatively low sedimentary organic matter load
compared to areas subjected to high density macroalgal mat, regardless of the amount of Hg present in the
sediments; (3) wind transport of plant masses in low energy storage areas, may constitute an increase factor of Hg
in the sediment. A critical review of the results was made in comparison with the available literature reports and
some hypotheses were proposed regarding the possible dynamics and availability of the contaminant.

1. Introduction
Mercury (Hg) is a contaminant that exists in different forms in
aquatic ecosystems (Harris et al., 2007) and the volatile monomethyl
mercury (MMeHg) is one of the most toxic form which acts as a
neurotoxin that may bioaccumulate in aquatic organisms and bio­
magnify through the food web, at the end of the pyramid, in fish and
then in their consumers (Castoldi et al., 2008). The greater part of Hg
introduced into the coastal sea is insoluble and falls in the sediment,
where it accumulates. Inorganic and organic Hg are bound to particles,
colloids and high molecular weight organic matter (Horvat, 1977).
Above all anoxic sediments favor MMeHg production by methylating
activity of sulphate-reducing bacteria (SRB) (Benoit et al., 1999, 2003;
Mason, 2002; Wiener et al., 2003; Fleming et al., 2006). MMeHg pro­
duced by bacterial activity can readily dissolve in the water column, can
be retained by mucoproteins and by lipids (Tremolada, 1998) and
accumulate in aquatic organisms. In the meat of fish species it is above
all MMeHg to accumulate at a rate of 60–90% compared to the total Hg
(Kannan et al., 1998). MMeHg and Hg(II) concentrations in fish can in­
crease by 6 orders of magnitude compared to the concentration present
in the water column (Chen et al., 2008), however, although many as­
pects of the accumulation processes are known (Pan and Wang, 2004),
there is probably still much to understand in relation to the dynamics of
massive accumulations in fish tissues. In fact, fish species behave
differently in the accumulation of Hg and may activate peculiar physi­
ological defense strategies. Recent studies have shown that the feeding
niche have a great impact on Hg accumulation, e.g., pelagic-feeding fish
resulted with higher MMeHg concentration compared to benthic-
feeding fish (Chen et al., 2009).
The mechanisms that control bioaccumulation depend a lot on the
sediment composition, since the binders present (e.g. organic matter,
OM; acid-volatile sulphides, AVS; metal oxides) are decisive in the
bioavailability of mercury. Some studies have shown an inverse corre­
lation between the amount of MMeHg released from the sediment and
the amount of particulate organic matter, indicating that OM binds Hg(II)
and MMeHg, retaining them in the sediment (Lawrence et al., 1999;

* Corresponding author.
E-mail addresses: lealab1@gmail.com (M. Lenzi), letizia.marsili@unisi.it (L. Marsili).

https://doi.org/10.1016/j.marpolbul.2020.111907
Received 11 August 2020; Received in revised form 24 November 2020; Accepted 30 November 2020
Available online 15 December 2020
0025-326X/© 2020 Elsevier Ltd. All rights reserved.
M. Lenzi et al.
McAlloon and Mason, 2003). On the other hand, OM is subject to
mineralization which in the eutrophic lagoon environment proceeds
essentially anaerobically up to over 80% of what is mineralized
(Jørgensen, 1983). Therefore, OM usually constitutes a transient binder.
Coastal lagoons, especially shallow-water, non-tidal lagoon, are
structurally eutrophic environments and tend to accumulate organic
matter in the sediment. The presence of Hg in such environments, even if
of mineral origin, commonly involves its mobilization as MMeHg; in
fact, the methylation process seems here to be more efficient than in
marine or river environments (Hurley et al., 1995).
The Orbetello lagoon (Tuscany, Italy) has a diffuse Hg anomaly, with
decidedly high concentrations in some areas. This lagoon and the whole
coastal strip of southern Tuscany are affected by detrital contributions
coming from the near Amiata Mount that is one of the most important
global sources of cinnabar. In particular, the seabed has homogeneous
total mercury values around 1.2 ng g− 1 (Abella et al., 2012). In the
Orbetello lagoon, some fish species are particularly subject to the
accumulation of Hg, beyond the allowed value for marketing (legal
limit), according to the Regulation 1881/2006/EC that defined 0.5 μg
g− 1 wet weight (w.w) for total mercury (T-Hg). This lagoon has also
been subject to eutrophication for some decades with the development
of huge blooms of opportunistic macroalgae which decay during the
summer season, leading to dystrophies, sometimes so extensive as to
cause die-offs of benthos and fish populations (Lenzi et al., 2011). These
phenomena are countered through some management interventions
such as the resuspension of the top-layer sediment (3–5 cm) with high
organic load. This process triggers aerobic mineralization of organic
matter, increases the sediment redox hindering bacteria sulphate
reduction activity, and, in cascade, a series of other environmental
mitigation processes take place (Lenzi, 2010).
The aim of this study was to shed light on some important aspects
concerning the bioaccumulation of Hg, a phenomenon that in sub­
merged coastal environments can take on very complex and often con­
tradictory features. To this scope, we had set ourselves the following
questions: 1) can the Hg contamination of fish occur in ways other than
through the food chain? 2) could the management of eutrophication
through the sediment resuspension favor and increase the release of
MMeHg in the water column?
2
Marine Pollution Bulletin 162 (2021) 111907
2. Materials and methods
This study was carried out through field surveys that involved the
sampling of abiotic and biotic matrices and in situ instrumental detec­
tion, and laboratory analysis to determine Hg in fish meat and in the
water column. In addition, a review of the studies carried out in this
lagoon on the water, sediment and fish Hg contents and on the load of
the sediment labile organic matter was conducted. Satellite images
coinciding with the periods examined in our study, relating to the
presence of high density macroalgal mat and the evidence of SRB ac­
tivity, were also used.
2.1. Study area
The Orbetello lagoon is a shallow, eutrophic coastal water body of
about 25.25 km2 in the southern Tuscan coast of Italy (42◦ 25′ –42◦ 29′ N,
11◦ 10′ –11◦ 17′ E). This lagoon has two communicating basins known as
West and East with an area 15.25 km2 and 10.00 km2 respectively, with
a 1 m mean depth (Fig. 1). Three artificial sea-lagoon canals, 0.5–3 km
long and 10–15 m wide, two in the West and one in the East basin,
connect the lagoon with the sea (Fig. 1; SLC 1, 2, 3). Because of the low
tidal excursions of the Tyrrhenian Sea and for the length and small width
of the three sea-lagoon canals, lagoon water turnover is poor and de­
pends mainly on wind force and direction. Lagoon salinity ranges from
28 to 45 (practical salinity scale), depending on rainfall and evaporation.
The environment is eutrophic due to fish-farm wastewaters, inter­
mittent streams containing agricultural run-off and civil effluent, and
historical input stored in the sediment (Lenzi et al., 2003). Due to high
nutrient availability, morphology and low-water-turnover, this lagoon is
subject to severe macroalgal proliferation, which can cause dystrophic
crises with die-offs (Lenzi et al., 2011). Macroalgal harvesting and sea-
water pumping were the first management actions taken to counteract
the eutrophication effects. Macroalgal harvesting occurred for 6 months
per year and took away from 4000 to 6000 t of wet macroalgal mass. It
was estimated that 6000 t of macroalgae removed from the ecosystem
involved the resuspension of over 15,000 t of sediments (Lenzi et al.,
2013). From 2014 to date, the action to counter eutrophication has
mainly been based on the only direct resuspension of the anoxic sedi­
ment top-layer (Lenzi et al., 2017). Sea-water pumping is the action still
Fig. 1. The Orbetello lagoon, located along the North
Tyrrhenian coast, divided into the two West and East
basins. SLC 1 and SLC2, sea-lagoon canals at SW and
Nord, respectively, of the West basin; SLC3, sea-
lagoon canal at the eastern end of the East basin;
blue circles, marine water pumping stations; squares
D, U1 and U2, areas of 9 ha each in which the DGTs
were placed, the first subjected to sediment distur­
bance and the other two left undisturbed; MA-1, 2, 3,
4 macroareas subjected to analysis of the sediment
Hg by Beccaloni et al. (2011). (For interpretation of
the references to color in this figure legend, the
reader is referred to the web version of this article.)
M. Lenzi et al.
practiced annually, between June and August, to promote water turn­
over, through an input flow coming from two western pumping stations,
at the lagoon-mouth of the two western canals (SLC 1, 2), and an output
flow by the eastern canal (SLC 3; Fig. 1).
2.2. Hg content in fish
Hg content analyses of the most marketed fish species were con­
ducted continuously from 2009 to 2019 (for the last year with only the
first six months available) by the Orbetello Pesca Lagunare Company
(OPL), the only fishing company that operates in the lagoon. The ana­
lysed fish came mostly from the fixed catching stations located at the
mouth of the sea-lagoon canals (Fig. 1) and to a lesser extent from
catches with trammel nets in areas close to the same mouths. The fish
catch station of the sea-lagoon canal SLC1 (Fig. 1) in the West basin
(West), is the one that produces more than 80% of the catch, especially
sea-bream (Sparus aurata L.). The catch areas have been divided overall
into West and East, in relation to the two lagoon basins. The subjects
caught for marketing at the fixed sea-lagoon stations are transferred to
network tanks pending the analytical results and only then they are
marketed. Since over the years it has been found that the subjects caught
in the East basin (East) had almost always values unsuitable for mar­
keting, fishing takes place mainly in the West and checks are only oc­
casionally carried out in the East to test the contamination state. For this
reason, the available data were 239 for West and only 30 for East, for a
total of about 1890 subjects sacrificed for the purpose.
Another 26 records came from other species over which an occa­
sional check is made. The species examined were: Anguilla anguilla L.,
Sarpa salpa L., Chelon labrosus Risso, Mugil cephalus L., Liza aurata Risso,
Dicentrarchus labrax L., Diplodus puntazzo Walbaum.
Hg content analyses were essentially focused on sea-bream, since this
species has become the most present in the lagoon (74% of the total
catches) due to the decrease of the other species since the 70s of the last
century, mainly due to the eutrophication increase and its consequences,
and the sowing of sea-bream juveniles for hundreds of thousands every
year (OPL, pers. comm.).
Each record came from the analysis of the homogenized meat of the
skinless back muscle of 4–10 subjects, in relation to the size of the stock
captured and stabulated for marketing. The subjects were divided into 4
classes in relation to weight: class1 < 300 g; 300 g = class2 < 400 g; 400
g = class3 < 500 g; 500 g = class4 > 500 g. The analyses were conducted
by the lab of the PH Company of Tavarnelle (Fi) using the Atomic Ab­
sorption Spectroscopy technique (MP 02/010) and expressed as total
Hg, in ng g− 1. The samples were finely homogenized and the analyzes
were performed in duplicate on sub-samples of 50–100 mg. The
analytical procedure was carried out directly on wet sub-samples, while
other sub-samples were dried at 105 ◦ C only to determine the water
amount. The results were calculated by comparison with a calibration
line obtained by a certified standard reference material. Measured and
certified values were at >95% confidence level.
2.3. DGT and experimental design
Diffusive Gradient in Thin films technique (DGT) was used to high­
light traces of metals in the water column. This technique developed in
the 90s of the last century (Davison and Zhang, 1994) and only more
recently it has been improved and also used for Hg (Clarisse et al., 2012;
Fernández-Gómez et al., 2011; Chakraborty et al., 2014; Wu et al.,
2017). DGT commercially available were obtained by ExposMeter
Sampling Tecnologies Co. (Tavelsjö, Sweden). The binding layer con­
sisted of a thiol resin embedded in a polyacrylamide gel, considered
suitable for measuring both organic and inorganic Hg species (more
information on devices in Fernández-Gómez et al., 2011).
The field experience was conducted in three plots of the lagoon
(Fig. 1), one of which was subjected to periodic sediment disturbance
(D) and two left undisturbed (U1 and U2). D and U1 were placed in West
3
Marine Pollution Bulletin 162 (2021) 111907
and U2 in East. Each plot was about 9 ha extended and delimited with
bamboo canes, so that the U were never crossed with boats, except at the
moment of data collection and sampling. Conversely, D was appropri­
ately marked so that a boat navigating inside it would produce sediment
soft top-layer (3–5 cm) resuspension, three times a week. DGTs were
placed in each plot, tied to a bamboo cane and kept in the middle of the
water column, i.e. about 60 cm from the surface. Each plot was simul­
taneously affected by 6 DGTs; they were divided into groups of three,
each placed in one half of the same area. Altogether, 18 DGT devices
were used for each survey. Three sampling surveys by DGT, from 15-
Sept 2014 to 23-Sept 2014, from 21-May 2015 to 31-July 2015, and
from 17-August 2016 to 19-September 2016 (group-1, group-2, group-3,
respectively), were conducted to detect T-Hg in the water column
(Fig. 1), for a total of 54 DGTs used. Temperature (T, ◦ C) and pH (by
electronic digital pHmeter DELTA OHM HD8705), salinity (S, practical
scale of salinity; by refractometer ATAGO S/Mill), and nephelometric
turbidity (NTU; by nephelometer by HANNA HI 93703-11 turbidity
meter) were measured in field during the surveys. Nephelometric
turbidity was measured only in non-disturbing conditions for both Us
and D, while in this last plot, the re-sedimentation time after the sedi­
ment resuspension action by boat was estimated for each intervention.
Total mercury content in DGT (Hg-DGT) was determined directly in
the DGT diffusive layer and resin gel after upper filter removal and
treatment with 1 M HNO3 elution solution. Selective mercury analyser
(Advanced Hg analyser, AMA-254, Altec, Czech Republic) based on
atomic absorption spectroscopy (AAS) was used. Hg-DGT was calculated
according to the following Eq. (1):
( )
Hg − DGT = CE VHNO3 + Vgel *Ef − 1 (1)
where: CE is the concentration of metals in the 1 M HNO3 elution so­
lution (in g L− 1); VHNO3 is the volume of HNO3 added to the resin gel;
Vgel is the volume of the resin gel; Ef is the elution factor for each metal,
typically 0.8.
The average concentration of Hg in the water column ([Hg], ng L− 1)
was calculated in relation to the hours of storage in water of DGTs, and
temperature and salinity of each dwell period, according to the
following Eq. (2):
(
[Hg] = Hg − DGT*0.094*1000* t*3.14*d*10− 6
)− 1
(2)
where: [Hg] is the Hg concentration in ng L− 1; Hg-DGT is the quantity in
Hg determined analytically in each DGT; 0.094 is the thickness of the
diffusive gel plus the thickness of the filter membrane, in cm; t is the
storage time in seconds of DGT; 3.14 is the exposure area; d * 10− 6 is the
diffusion coefficient of metal in the gel (variable according to temper­
ature and salinity values and obtained by specific table for open pore gel;
the numbers need to time 10− 6 and the unit should be cm2 s− 1).
2.4. Statistical analysis
The different level of mercury contamination in fish and water in
relation to different groups of analysis (weight class for fish, disturbed
and undisturbed site for water) were assessed by non-parametric tests
that being distribution-free procedures no assumptions about the
sampled population are requested (Corder and Foreman, 2014) and
generally are more robust to outliers. In particular, the Mann-Whitney U
Test and a Kruskal-Wallis equality of populations rank test were used to
evaluate statistically significant differences between groups defined on
the basis of two or more unrelated groups (Hollander et al., 2014).
Specifically, we test the null hypothesis that the population centres
are all equal against the alternative hypothesis that the other population
(at least one of the populations in the Kruskal–Wallis test) tends to
exhibit larger values than the baseline population (at least one of the
other populations in the Kruskal–Wallis test). We also performed mul­
tiple pairwise comparison tests using the Holm-Sidák adjustment (Holm,
M. Lenzi et al. Marine Pollution Bulletin 162 (2021) 111907

1979).
In order to validate the use of a non-parametric approach pre­
liminary analyses were also conducted for checking normality of data
and the presence of extreme values under study. Visual inspection of the
empirical frequency distributions clearly showed lack of symmetry
(positive skewness) and the Shapiro-Wilk test gave p-values less than
0.05 for the outcome variable (Peat and Barton, 2005). Moreover, box
plots have been used to identify extreme values of each variable by
groups. In these box plots, any data that lies outside the upper or lower
fence lines is considered a possible outlier.
All statistical analyses were performed using STATA 14 software
(StataCorp, 2015).
3. Results
3.1. Hg content in fish
Sea-bream Hg values (μg g− 1 wet weight) coming from catches in
West and East, are reported in Tables 1 and 2, respectively. Since these
are random determinations on stocks awaiting marketing, class-2 and, to
a lesser extent, class-3 are those with the largest number of subjects
examined (overall 87% of records in West, and 86% in East).
Among all West samples (n = 239), the mean was 0.30 ± 0.23 μg g− 1
and the median 0.22 μg g− 1. 82.84% of the samples resulted within the
limit 0.5 μg g− 1 allowed for the marketing, among these classes 1 and 2
together were below the limit in 81.54% of the cases, classes 3 and 4
together in 85.92% of the cases. With regard to the 30 records from East
(of which 3 in class-1, 20 in class-2, 6 in class-3 and 1 in class-4), the
overall mean was 0.78 ± 0.44 μg g− 1, the median 0.81 μg g− 1 and the
range 0.05–1.73 μg g− 1. Means and medians were all beyond the legal
limit. 8 samples (26.67%) were within the limit allowed for commer­
cialization, all in class-2, which however was the largest.
In West, the highest medians were obtained for class-2, in East, the
highest median was obtained in class-1. Hg ranges have been widely
variable in both basins, in particular in the West (Tables 2, 3).
Fig. 2 gives a quick visual assessment of sea-bream meat Hg con­
centration (μg g− 1 wet weight) from catches in West e in East. The box
plots show the median Hg concentration was higher in East area, and
that West area was characterized by some values that were outside the
expected range. Mann-Whitney test results confirmed a significant dif­
ference between the two areas (p < 0.0001).
In order to obtain a reliable sample size, we merged observations of
class-1 and class-2 (1 + 2) and class- 3 and class-4 (3 + 4) in both areas.
In Fig. 3, Hg concentration boxplots using these super-groups, in West
and East areas are reported. Mann-Whitney test results showed a sig­
nificant difference between the two super-groups in West, whereas the
difference is not significantly different in East (p = 0.6834).
Using Kruskal–Wallis test, we also tested for significant differences
across years in West (p < 0.0001). Findings show significant differences
across years. The same test is not reported for East area because the
sample size is too small in some years.
In Table 3, Hg contents for other fish species are reported, together
with the number of subjects used for analysis and their mean weight,
distinguishing the origins from West and East. For West A. anguilla,
D. labrax and mullet and for East D. puntazzo, the means (± SD) are
reported.
Species coming from the East were more impacted, but among these
C. labrosus accumulated the least amount of Hg, even for subjects with
relatively large dimensions. D. puntazzo accumulated more Hg, in both
West and East, together with D. labrax; in West, the three species of
mugilidae examined and S. salpa were the organisms with the least
accumulation of Hg. A. anguilla presented variable values in the West,
but always above the legal limit in the East.
3.2. DGT
In Table 4, the calculated values of the mercury concentration in the
water column ([Hg], ng L− 1) and temperature, pH, salinity and nephe­
lometric turbidity values measured in the plots during the experience,
and the residence time of the DGTs in hours in D, U1 and U2, are re­
ported. The re-sedimentation time after the sediment resuspension by
boat in the plot D was on average 18.7 ± 5.6 min.
Hg means of the three plots for each trial showed markedly higher
values in U2 for 2014–15, while they fell homogeneously for all three in
2016. The means among all the records were 6.55 ± 2.88 ng L− 1, 7.95 ±
5.55 ng L− 1, 9.94 ± 2.89 ng L− 1, per D, U1 and U2, respectively. In

Table 1
Mean (±SD), median and range of Hg concentration (μg g− 1) and number of samples analysed (n) in muscle of sea-bream 4 age classes (class1 < 300 g, 300 = class2 <
400 g, 400 = class3 < 500 g, 500 = class4 > 500 g) caught in the West basin of the Orbetello lagoon, between 2009 and 2019.
Class 1 Class 2 Class 3 Class 4

n Mean Median Range n Mean Median Range n Mean Median Range n Mean Median Range

0.16 ± 0.12 ±
2009 10 0.24 0.06 0.03–0.79 4 0.11 0.08 0.03–0.30
0.13 ± 0.12 ± 0.09 ±
2010 1 0.08 9 0.05 0.14 0.07–0.91 3 0.02 0.13 0.1–0.14 2 0.07 0.09 0.02–0.16
0.25 ± 0.23 ±
2011 7 0.08 0.23 0.15–0.37 2 0.01 0.23 0.24–0.23
0.33 ±
2012 17 0.23 0.24 0.08–1.08 1 0.99
0.35 ± 0.19 ±
2013 2 0.15 0.35 0.21–0.50 3 0.13 0.10 0.10–0.38 1 0.37
0.16 ± 0.22 ± 0.18 ±
2014 7 0.07 0.16 0.06–0.30 6 0.08 0.22 0.10–0.32 3 0.01 0.18 0.17–0.19
0.34 ± 0.37 ±
2015 1 0.15 26 0.14 0.30 0.06–0.84 10 0.21 0.34 0.16–0.69 1 0.50
0.31 ± 0.23 ± 0.25 ±
2016 1 0.09 20 0.18 0.31 0.09–0.73 3 0.15 0.27 0.03–0.79 2 0.02 0.25 0.23–0.26
0.26 ± 0.25 ± 0.26 ±
2017 17 0.19 0.24 0.05–0.83 14 0.24 0.13 0.06–1.03 3 0.17 0.17 0.12–0.50
0.41 ± 0.42 ± 0.17 ± 0.30 ±
2018 6 0.21 0.4 0.18–0.74 33 0.27 0.38 0.06–1.07 5 0.09 0.19 0.06–0.21 3 0.13 0.38 0.11–0.40
0.26 ± 0.49 ± 0.25 ±
2019 3 0.06 0.22 0.22–0.34 7 0.25 0.46 0.14–0.91 6 0.29 0.13 0.06–0.82
0.30 ± 0.31 ± 0.26 ± 0.22 ±
Total 12 0.20 0.22 155 0.23 0.24 53 0.22 0.17 19 0.14 0.18

4
M. Lenzi et al. Marine Pollution Bulletin 162 (2021) 111907

Table 2
Mean (±SD), median and range of Hg concentration (μg g− 1) and number of samples analysed (n) in muscle of sea-bream 4 age classes (class1 < 300 g, 300 = class2 <
400 g, 400 = class3 < 500 g, 500 = class4 > 500 g) caught in the East basin of the Orbetello lagoon, between 2009 and 2019.
Class 1 Class 2 Class 3 Class 4

n Mean Median n Mean Median Range n Mean Median Range Record

2009 1 0.05
2010 5 0.67 ± 0.23 0.72 0.33–0.91 1 0.23
2011 5 0.83 ± 0.58 1.26 0.10–1.33 1 0.76
2012 3 0.79 ± 0.34 0.93 0.32–1.12
2013 1 0.16
2014 3 0.67 ± 0.26 0.49 0.40–1.04 2 0.96 ± 0.26 0.96 0.70–1.22
2015 1 0.84 1 0.67 1 0.45
2017 1 1.52 1 0.92 0.79
2018 1 1.73
2019 1 1.32
Total 3 1.36 ± 0.38 1.52 20 0.67 ± 0.43 0.63 6 0.71 ± 0.32 0.73

Table 3
Hg concentration (μg g− 1) in fish muscle of 7 fish species, number of samples analysed (n), mean
weight (±SD) of the subjects analysed in the East and West basins of the Orbetello lagoon, between
2009 and 2019. In gray, the values of which the Hg mean (±SD is in bold).
Species Year n Body weight West East
g Hg ( g g−1) Hg ( g g−1)
A. anguilla 2009 7 163 ± 5 0.513
2010 6 160 ± 29 0.321
2011 8 210 ± 32 0.131
2012 7 150 ± 4 0.553
2015 11 230 ± 4 0.845
2015 10 150 ± 3 0.420
0.346±0.142
S. salpa 2011 6 410 ± 33 0.023
C. labrosus 2014 4 855 ± 41 0.166
2017 4 660 ± 24 0,131
2018 3 1907 ± 106 0.256
2019 3 1063 ± 101 0.130
M. cephalus 2010 3 940−37 0.010
2012 4 717 ± 25 0.055
L. aurata 2010 12 261 ± 8 0.092
2012 10 240 ± 5 0.220
2013 10 320 ± 5 0.063
2017 8 270 ± 11 0.071
2018 10 332 ± 12 0.025
0.089±0.061
D. labrax 2011 3 507 ± 39 0.447
2013 4 593 ± 56 0.733
2017 3 553 ± 21 0.780
2018 3 723 ± 27 1.100
0.776±0.267
D. puntazzo 2011 6 300 ± 22 1.940
2013 4 350 ± 19 0.487
2014 7 450 ± 32 1.200
1.209±0.593
2019 9 324 ± 4 0.769

Fig. 4, Hg concentration (ng L− 1) boxplots by plots are reported over
years. The median of Hg concentration in U2 was higher than in D and
U1 plots, and Kruskal–Wallis statistics showed significant differences
between the three plots (p = 0.0005) only in 2015. We also performed
multiple pairwise comparison tests. Statistical difference compared to D
was observed only for U2 (p = 0.0001), whereas Hg concentrations
significantly differed between U1 and U2 (p = 0.0257).
The annual means among the records of all three plots were 11.48 ±
4.47 ng L− 1, 6.35 ± 3.10 ng L− 1 and 6.60 ± 2.63 ng L− 1, for 2014, 2015,
2016, respectively.
3.3. Satellite imagery
Some satellite images of Sentinel-2 by LandViewer Web page, of the
2018–19 of Orbetello lagoon, relating to the distribution of macroalgal
mats and the development of dystrophic phenomena with evidence of
sulphur-blooms, have been used for the discussion of data.
4. Discussion of the results and comparison with previous
studies
4.1. T-Hg in the water column and the importance of the sediment
The residence time of DGTs in the water consistently increased be­
tween 2014 and 2015, from 8 to 32 days, and also increased between
2015 and 2016, from 32 to 72 days. The DGTs were left in the water for a
longer period of time in order to detect mercury releases from the
sediment. Even if there was a difference in the residence times of the

5
M. Lenzi et al. Marine Pollution Bulletin 162 (2021) 111907

[Hg], mercury concentration (ng L− 1) and mean (±SD) in bold, T, temperature range (◦ C), S, salinity (PSS), pH, nephelometric turbidity (NTU) and t, time in hours the DGT devices stay in the water between 2014 and 2016

7.6 ± 2.4

5.4 ± 2.2

6.8 ± 2.7
[Hg]

10.1

10.1

10.0
5.2

5.5
9.9
4.9

7.6
3.2
8.0
2.6
7.0
4.0

9.6
4.1

3.3
5.0
8.9
NTU

1.92

1.88

2.00
8.60

8.60

8.40
pH

37–38

37–38

37–38
pH

25–29

25–29

25–30
(DGT groups in the trials 1, 2, 3), in the stations D (disturbed area in the West basin), U1 (undisturbed area in the West basin) and U2 (undisturbed area in the East basin).

768

768

768
Fig. 2. Box plots of Hg concentration (μg g− 1 wet weight) found in sea bream

T
meat in West and in East basins of the Orbetello lagoon.

Aug-Sept

Aug-Sept

Aug-Sept
Group-3
2016

10.5 ± 0.7
3.2 ± 0.2

5.4 ± 0.4
[Hg]

10.0
11.0

11.7
10.7
3.0
3.4
2.9
3.5
3.2
3.2

5.4
5.4
4.8
5.7
4.9
6.0

9.9

9.7
NTU

1.72

1.71

1.83
8.40

8.40

8.60
pH

34–38

34–38

33–38
S

19–35

19–35

19–35
Fig. 3. Box plots of Hg concentration (μg g− 1 wet weight) found in Class 1 + 2
T

and Class 3 + 4 of sea-bream by West and in East basins of the Orbetello lagoon.
1728

1728

1728
DGTs between the trials, the aim of the trials to make a comparison
t

between the plots within each period, one of which subjected to sedi­
May-July

May-July

May-July
Group-2

ment resuspension, was accomplished. No significant difference was

2015

found between the plots, except in May–June 2015, for higher values in
U2, before a severe dystrophic crisis in East basin.
However, the recent study by Díez and Giaggio (2018) showed that a
13.1 ± 6.9

12.5 ± 0.6
8.9 ± 1.2

prolonged residence time of the devices in the water over 10 days, and
suspended particulate could compromise the results, producing un­
[Hg]

10.0

10.3

20.0
18.9
21.0

13.0
12.0
11.6
13.1
12.1
13.2
7.7

9.5
8.4
7.2

6.2

5.5
7.0

derestimations. Therefore, that study questions the results of our study.

Nevertheless, we considered the results to be still useful because of the
NTU

2.21

2.21

2.27

following three reasons: 1) the lagoon water in the examined periods

was transparent, as shown by the nephelometric data (Table 4), a
common condition in water overlying macroalgal mats due to compe­
8.40

8.40

8.45
pH

tition with microalgae (McGlathery et al., 2007); so we can assume that

the amount of suspended material was likely moderate; 2) sediment
resuspension conducted in plot D brought into the water column, coarse
38

38

39
S

organic particulate, produced by the decay of the macroalgal under-

layer mat, which sedimented in 15–20 min, then the water became
24–28

24–28

24–28

transparent again; 3) our data were substantially similar to those

T

detected by a parallel survey carried out between 2014 and 2015 by the
Regional Environmental Protection Agency of Tuscany (ARPAT, 2016;
192

192

192

Report for the Institutions) in the same plots D, U1 and U2. ARPAT is the
t
Group-1

official Italian institute appointed to the monitoring in the Water

2014

Sept

Sept

Sept

Framework Directive.
The ARPAT study revealed a very high total Hg content in the water
Table 4

column in June 2014 (940–183 ng L− 1). However, it declined in August-

U1

U2
D

6
M. Lenzi et al. Marine Pollution Bulletin 162 (2021) 111907

modified carbon nanoparticles SH-CNPs suspension as the liquid binding

phase and cellulose acetate membrane as the diffusive layer), observed
that the greatest efficiency in contaminant grabbing was obtained at pH
close to neutrality.
In the sediment, the study of ARPAT (2016) found high values of T-
Hg in all plots, always above AA-EQS. (0.3 μg g− 1 dw; Ausili et al.,
2018), comprised between 0.76 μg g− 1 dw and 2.80 μg g− 1 dw (Table 5),
and established that there were no significant differences among the
data populations of the three plots, neither before nor after the treat­
ment of D through sediment resuspension.
In 2008, a study conducted by ICRAM (today Italian Institute for
Environmental Protection and Research, ISPRA) and University of Siena
established ranges of 0.56–0.95 μg g− 1 dw for West and 0.25–6.04 μg
g− 1 dw for East, for the sediment 5 cm top-layer (ICRAM-UNISI, 2008;
Report for the Institutions), with the highest values detected in the
westernmost part of the East basin.
Fig. 4. Box plots of Hg concentration (ng L− 1) by the DGT stations D Similarly, a survey by Beccaloni et al. (2011) (Institute of Health, ISS;
(disturbed) and U1, U2 (undisturbed), in 2014, 2015 and 2016. D and U1 in Report for the Institutions) confirmed high values in the East sediment,
West basin, U2 in East basin. highlighting 4 different macro areas: macroarea 1, near the town of
Orbetello, with values between 0.19 and 0.94 μg g− 1 dw; macroarea 2 at
2014 (about 55–96 ng L− 1), November-2014 (10–18 ng L− 1) and April- the east end, with a range 2.1–21.7 μg g− 1 dw; macroarea 3, close to the
2015 (<5 ng L− 1) until it reached in November values well below the sand dune, with range 0.17–0.43 μg g− 1 dw; macroarea 4, in the far west
maximum allowed concentration of 70 ng L− 1 (MAC- EQS; Ausili et al., of the basin, close to an old mining area, with range 3.8–11.72 μg g− 1 dw
2018) (Table 5). The trend observed by ARPAT confirms the results of (Fig. 1).
our DGTs for T-Hg, which had shown a decreasing trend between 2014 All these studies showed a wide variability of the sediment data, the
and 2016; furthermore, the ARPAT’s T-Hg concentrations of the three highest Hg concentrations in the East, and in both East and West the
plots did not show significant differences in each of the observation overcoming of the EQS value.
periods. The difference between September-2014 DGT results and Beyond the Hg concentration in the sediment, i.e. of its contamina­
ARPAT’s June-2014 and August-2014 results was about 1 and 0.5 order tion potential, DGTs results and the ARPAT study highlighted different
of magnitude, respectively. dynamics of the releases from the sediment, between one year and
Putting together our data and ARPAT’s ones the decreasing trend another and between months of the same year. This could be due to
between 2014 and 2015 was confirmed. Our data of September-2014 are changes in environmental conditions, especially as a function of the
inserted temporally between the ARPAT results of August and accumulations of labile organic matter (LOM) available for anaerobic
November, very close to November. In 2015, the DGTs means varied mineralizers.
between 3 and 10 ng L− 1, and ARPAT values were below 5 ng L− 1 SRB act on the mineral deposits of the cinnabar (HgS) and on the
(Tables 4 and 5). organic compounds sulphurated (HgS(SH)− , Hg(SH)2, Hg(Sn)HS− )
However, it is important to take into account other possible criti­ containing Hg, methylating Hg to MMeHg. SRB are active in a range of
calities of the DGTs data: 1) Hg levels may have been underestimated environments from micro-oxidized to anoxic. Sediment oxygenation
because of the presence of a high quantity of dissolved organic matter with low LOM load decreases methylation (Benoit et al., 2006). In fact,
(DOM): Hg-DOM complexes bind a large part of HgII, competing with to undertake sulphate-reducing activity and also to start methylation,
the capture by DGT (Fernández-Gómez et al., 2011). Previous studies SRB require at the same time low or zero oxygen and a significant load of
demonstrated relatively high and homogeneously diffused values of LOM in the sediment. Therefore, the presence of oxygen and low OM
DOM in West (Lenzi et al., 2015) and East basin (Lenzi et al., 2019); content in the sediment lead to no or very low SRB activity. When the
these DOM values could have contributed to the underestimation of Hg, sulphate reducing activity becomes predominant, the concentration of
but probably did not invalidate the comparison between the three plots; sulphides increases, so as to inhibit methylation (Benoit et al., 1999).
2) during the DGTs residence time, the pH values recorded in the lagoon According to the conceptual model reported in Fig. 5, we can hypoth­
plots were relatively high. Wu et al. (2017), using similar devices (thiol- esize a range of environmental conditions starting from an extreme of
oxidized sediments, in which the sulphate-reducing activity is scarce or
zero and so is the methylation, up to the opposite extreme situation of an
Table 5 anoxic/dystrophic state with very intense sulphate-reducing activity,
Hg values estimated in the water and sediment of the Orbetello lagoon, in the
accumulation of acid-volatile sulphides and little or no methylation.
plots D, U1 and U2, between June 2014 and April 2015, by the Regional Envi­
Along a growing gradient of sulphate-reducing activity, between the two
ronmental Protection Agency of Tuscany (from ARPAT, 2016; reconstructed).
extreme conditions, the methylation increases reaching its peak more or
T-Hg water T-Hg sediment
less centrally between the two conditions described.
June-14 Aug- Nov- Apr- June- Aug- Nov- Apr- In Fig. 6, the LOM trend in the West is reported between 2014 and
14 14 15 14 14 14 15 2016. LOM decreased between June 2014 (20.78 ± 3.33%) and June
ng L− 1
ng L− 1
ng L− 1
ng μg μg μg μg 2015 (8.79 ± 2.54%) probably due to artificial sediment resuspension
L− 1 g− 1
g− 1
g− 1 g− 1
(Lenzi et al., 2017). In the East, LOM in July 2014 measured 16.11 ±
D 127.60 70.00 14.00 <5 1.46 1.42 1.94 1.92 9.49% (unpubl. data), in June-2015 15.07 ± 8.81% (sampling con­
± 28.34 ± 4.24 ± 2.53 ± ± ± ± ducted in the easternmost part of East; unpubl. data) and in July–August
0.22 0.37 0.49 0.49 2015 there was an intense dystrophic event extended to the whole East
81.00 1.87 1.13 1.55 1.73
117.20 ± 11.20 ± ± ± ±
basin (Lenzi et al., 2016).
U1 ± 38.24 10.49 ± 2.40 <5 0.58 0.49 0.39 0.46 Based on these observations we can hypothesize that: 1) the high Hg
74.00 1.88 1.68 2.33 2.05 values in the 2014 water column in the two basins could be due to SRB
137.20 ± 12.20 ± ± ± ± activity supported by abundant LOM, probably started in the spring
U2 ± 17.35 11.78 ± 2.93 0.28 0.16 0.13 0.41
<5
phase, when the sulphide concentration was not high enough to inhibit

7
M. Lenzi et al.
Fig. 5. In the diagram (concentration, along the ordinates; variable environ­
mental conditions, along the abscissas), hypothetical sediment environmental
situations are depicted, from the oxidized state on the left, to the increasingly
reduced and anoxic state, proceeding to the right on the abscissas. The trend of
the redox potential (Eh, in mV; black line) goes towards increasingly negative
values (from left to right) with the gradual decrease in dissolved oxygen, in
conjunction with the increase of organic matter load in the sediments (OM;
brown line). As this situation proceeds towards anoxia/dystrophy, the activity
of sulphate-reducing bacteria increases, thus increasing the concentration of
acid volatile sulphides (AVS; yellow line). The sulphate reducing bacteria
operate the methylation of the Hg present. This last process increases with the
bacterial activity increase, until the AVS concentration becomes excessive and
inhibits the process (trend of the gray line). Therefore, two moments are
identified in which the concentration of MMeHg is the lowest or zero (blue
circles), and an intermediate phase in which this concentration is at the highest
values (red circle), coinciding mainly with the intermediate seasons. (For
interpretation of the references to color in this figure legend, the reader is
referred to the web version of this article.)
Fig. 6. Distribution of the LOM data transformation along the ordinates for
each trial, in West basin sediment, between June 2014 and September 2016
(from Lenzi et al., 2017).
the methylation process; 2) artificial LOM reduction in West would have
consequently reduced the SRB activity between 2015 and 2016, and
therefore Hg methylation; 3) persistence of high LOM values in the East
would have led to the 2015 summer dystrophy, reducing methylation
due to excess of AVS; thus, the DGT showed lower T-Hg values, which
were maintained in 2016 due to the drastic LOM decrease caused from
the dystrophy itself. This hypothesis places the West and East basins at
the two extremes of the model proposed in Fig. 5, for the period
2015–16.
8
Marine Pollution Bulletin 162 (2021) 111907
4.2. Fish contamination
Results of many previous studies confirm the East sediment is a
source of Hg contamination greater than the West one. Although the
contaminated fish can move freely between East and West basins, also
the West sediments represent a source of contamination, as highlighted
by DGTs and ARPAT (2016). However, the higher contamination of the
smaller sea-bream classes compared to the larger ones in the West,
contrasts with the model that foresees an increase of the contaminant
with increasing size of fish, caused by the tissue bioaccumulation
through feeding (Boening, 2000).
Miniero et al. (2013), in a study limited to sea-bream captured in the
East basin of the Orbetello lagoon, did not detect differences in the level
of contamination nor in size of the subjects examined, nor differences
between populations that could be attributed to area-specific feeding
patterns, which was also confirmed by our results in the East, although a
relatively small number of samples were used in both the study cited (n
= 26) and ours (n = 30). Miniero et al. (2013) estimated Hg content
between 0.36 and 1.58 μg g− 1, with minimum values higher than ours,
and a median of 1.13 μg g− 1, higher than ours.
Data variability for all sizes, especially in the West, does not exclude
an area-specific food bioaccumulation, but does not exclude contami­
nation may have occurred due to contact with the MMeHg released from
the sediments as a consequence of the SRB activity. Perhaps both events
have occurred; however we would like to focus on the release of
contaminant due to the SRB activity, which occurs more or less intensely
every year, and is made evident above all for the consequences of the
anoxia induced in the sediments by the macroalgal masses, in both West
(Fig. 7, a, b) and East, the latter particularly affected in macroareas 2
and 4 (Fig. 8, a, b). In the spring, before the sulphide accumulations
increase extremely producing dystrophic evidence (Figs. 7, 8, c, d), the
SRB activity could be the cause of intense releases of MMeHg.
Since DGT devices showed that Hg can be found in the water column
and can accumulate, over time, in membranes that contain diffusive and
binding gel, it can be assumed that contact contamination may also
occur in fish. Hg releases from the sediment would occur in different
areas, at different times, and could affect all vagile organisms. The
production of MMeHg would be all the more important, the greater the
deposit of the contaminant in the sediment (Beccaloni et al., 2011), but a
deviation from this logic could occur according to the SRB activity, as
can be observed from the conceptual model of Fig. 6.
Beccaloni et al. (2011) established that the abundance of reducing
sulphite bacteria in the sediment was greater in East macroareas 1 and 3,
i.e. those that had relatively lower Hg values. This could mean that
bacterial pool was relatively small in macroareas 2 and 4 due to high
levels of contaminant. However, macroareas 2 and 4 are also those
subject to the greatest accumulations of algal masses, so we cannot
exclude that accumulation of acid- volatile sulphides resulting from the
sulphate-reducing process have inhibited the SRB development, the
production of MMeHg and other bacterial strains including sulphite
reducers. Thus, paradoxically, the areas with a relatively lower level of
contaminated sediments, which however do not undergo excessive
accumulation of algal masses, such as to trigger an intense production of
sulphides, could be more productive in terms of MMeHg.
Sea-bream are the most abundant organisms marketed in this lagoon
and it can be assumed that the subjects of small sizes proceed in arrays of
more individuals and are more mobile than those of larger dimensions,
more sedentary, territorial and tendentially solitary (Froese and Pauly,
2004). Therefore, the small sizes may incur a more frequent MMeHg
release, while the larger sizes would be affected if and when the releases
affect their territory. All this would take place in a complex context of Hg
accumulation by food. Miniero et al. (2013) also came to similar
conclusion, claiming “.. it is possible that the high Hg found in fish from
the Orbetello lagoon depend not only on metabolism but also on the
proximity of the source: this, in turn, would have some bearing on the
lack of a relationship between Hg concentrations and fish size”. Where
M. Lenzi et al. Marine Pollution Bulletin 162 (2021) 111907

Fig. 7. The Orbetello lagoon West basin. The dark part in “a” and “b” consists of a vast macroalgal mat, in spring images, which, year after year, affects the central
western area; the whitish patches in “c” and “d”, more or less in the same areas as the algal mat, are the formation of colloidal sulphur produced in the water column
by the oxidation of hydrogen sulphide from the summer SRB activity in the same years as “a” and “b”.

the source is the lagoon sediment.
The hypothesis shows a weakness if we take into account the mullet,
which would seem not particularly predisposed to bioaccumulation of
the contaminant, neither through feeding, due to its low position in the
trophic level, nor through contact. A. anguilla also shows relatively low
Hg levels, especially in the West, despite having a predator diet, albeit at
a still relatively low trophic level, and a living condition that sees it
submerged in the mud, where the contaminated could be more present
in interstitial waters.
An explanation of this apparent anomaly could be the abundant
presence of skin mucus produced by both mullet and eels, compared to
sea-bream, sea-bass and D. puntazzo. Mucus is a colloidal, viscous sub­
stance that contains antibacterial enzymes and glycoproteins with high
molecular weight (mucins), often acidic by sialic acid (a carboxylated
monosaccharide) or sulphated monosaccharides (Dash et al., 2018). It
could represent a storage vehicle for Hg, a sort of filter that reduces the
crossing of the epidermis by the contaminant. The mucus would be
easily lost by the fish during the rubbing in the mud, between the algae
and the eelgrass meadows, and subsequently renewed. Therefore,
further studies are needed to shed light on species-specific differences in
fish Hg bioaccumulation, related to the skin protective mucus.
4.3. Vegetable masses as a vehicle for the accumulation of sediment Hg
ICRAM-UniSi (2008) established for macroarea 4 (Fig. 1) a high
variability of Hg content along 1 m sediment cores, with extremely high
peaks even in the deep layers. This was not the case for the other mac­
roareas and not even in the West, where Hg was located above all in the
first 20 cm. This suggests that there may have been a geological reason
(mining area) for macroarea 4 that led to the accumulation of a large
amount of Hg in the deeper levels, compared to all the other areas.
However, the abundance of this metal in the surface sediment in mac­
roarea 2 and other lagoon areas cannot be explained in the same way,
therefore there must be other origin of mercury accumulation.
In the Est basin, algal masses, both those locally produced and those
transported by the wind, accumulate more precisely in macroareas 2 and
4 (Fig. 8a,b). The cause of the high Hg level in the sediment top-layer of
the two macroareas could be due to the transport of the algal masses and

9
M. Lenzi et al.
Fig. 8. The Orbetello lagoon East basin. The celestial lines in “a” and “b” highlight the accumulations of algal masses at the two ends of the basin; the yellow lines in
“c” and “d” show the effects of dystrophic events located at the two ends of the basin in the summer of the same years as “a” and “b”. The lines contain macro areas 2
and 4, respectively in the far east and far west of the basin. (For interpretation of the references to color in this figure legend, the reader is referred to the web version
of this article.)
the phanerogams fronds that detach in autumn. Coelho et al. (2009), for
the lagoon of Ria de Aveiro (Portugal), demonstrated in fact a phe­
nomenon of Hg enrichment in areas far from sources precisely as a
consequence of the transport of macroalgae and phanerogam fronds
from areas contaminated by anthropic activity. They estimated Hg
accumulation in the fronds of Zostera noltii of 0.017–0.590 μg g− 1, and in
the thalli of Enteromorpha sp. up to a maximum of 2.1 μg g− 1. The
ICRAM-UniSi (2008) study established ranges of Hg accumulations for
algae and phanerogams of 0.020–0.040 μg g− 1 and 0.070–0.320 μg g− 1
in West and East, respectively. Therefore, the transport of the masses by
the wind could actually constitute a vehicle for the accumulation of Hg
and its redistribution in the sediment.
Considering that the morphology of Orbetello lagoon East basin has
been substantially unchanged for centuries, it seems plausible that, at
least for macroarea 2, the accumulation of vegetable masses may have
increased, over time, the Hg levels in the sediment by a factor of 20 to
40, compared to the bottom of the nearby sea, which is also affected by
relatively high levels of Hg due to debris transport from Amiata Mount.
The hypothesis opens up a new perspective of study and management of
critical issues on this basin.
4.4. Sediment resuspension and Hg release
Sediment resuspension in shallow water lagoon is a relatively
frequent process. A wind of intensity >8 m s− 1 can produce a distur­
bance in the sediment top-layer up to about 1.5 m in height of the water
column (Rubegni et al., 2013). Resuspension effects of the anoxic top-
layer sediment (for about 3–5 cm of thickness) have long been studied
for long time, observing numerous benefits in countering the eutrophi­
cation consequences, when disturbance is produced with high fre­
quency, including the reduction of OM load (Lenzi et al., 2010;
Martelloni et al., 2016).
Although OM is a binder for mercury and is considered crucial for its
bioavailability, OM is predominantly subjected to anaerobic bacterial
decay. Gill et al. (1999) and Mason et al. (2006), in fact, claimed the
flow of MMeHg from the sediment was not significant in their
10
Marine Pollution Bulletin 162 (2021) 111907
experiences, except under conditions of poor oxygenation, hypoxia-
anoxia, and in low redox values of the sediment top-layer. LOM,
therefore, does not produce a burial of Hg in the sediment layers since it
is an ephemeral blocker. Once released during OM decay, HgII and
MMeHg can affect the water column through a diffusion gradient.
Furthermore, the natural disturbance processes (wind, bioturbation,
bio-irrigation) favor a partial OM oxidation and mineralization, with the
consequent possible MMeHg release, which anyway doesn’t signifi­
cantly stop the SRB activity, therefore not influencing the production
process of MMeHg; indeed SRB presence in micro-niches with low ox­
ygen rates and their potential role in Hg speciation have been demon­
strated (Compeau and Bartha, 1985; King et al., 2001).
During sediment resuspension, iron and manganese sulphides are
rapidly oxidized in the water column, and the metal ions released
(Simpson et al., 1998; Caetano et al., 2002); on the contrary, Hg and Pb
are more difficultly released by their sulphides (Caille et al., 2003).
Furthermore, the rapid releases of Fe and Mn from the respective sul­
phides result in equally rapid precipitations of the corresponding oxides
and hydroxides, which can quickly adsorb the cations of the other heavy
metals released from their sulphides (Saulnier and Mucci, 2000; Caetano
et al., 2002) due to the huge abundance of Fe and Mn compared to trace
metals. In addition, in the sediment anoxic top-layer, some heavy metal
cations, including Hg, can take over into the Fe and Mn monosulphides
forming an even more insoluble sulphide that does not release cations
even during resuspension events (Di Toro et al., 1990; Allen et al., 1993).
These studies may explain why, with the DGTs and through the
parallel study of ARPAT (2016), the resuspension of the top-layer sedi­
ments carried out in the experimental area D, did not increase the
bioavailability of Hg compared to the non-disturbed areas, even in 2014
when the highest T-Hg values were found in the waters of both lagoon
basins.
Even if a three years study is relevant, given the danger that MMeHg
entails, further confirmation are required in order to ascertain the
repeatability of the results, repeating the experience with DGTs taking
into account as much as possible environmental criticalities, such as pH
and DOM, and reducing the time spent in water by the devices to remove
M. Lenzi et al.
doubts about any underestimation, and verify the real possibility of
using DGTs in lagoon environment.
CRediT authorship contribution statement
The paper has been carefully reviewed by the corresponding author.
Mauro Lenzi coordinated the work in the field and the drafting of the
paper.
Enrica Franchi collaborated in the field coordination and data
acquisition of the OPL.
Leporatti-Persiano worked in the field with the DGT and water
quality measurements.
D’Agostino and Gennaro took care of statistics.
Marsili collaborated in the drafting of the text.
Declaration of competing interest
This research work took place in total autonomy, without external
funding, nor through personal relationships of each of the authors with
external funding bodies.
Acknowledgment
We thank the Orbetello Pesca Lagunare Company for allowing the
use of Hg analysis results in pre-marketed fish, and for making rooms
and boats available.
References
Abella, J.A., Baino, T.R., Serena, F., Voliani, A., 2012. Evaluation of the Concentration of
Some Pollutants in the Sole (Solea solea) and State of the Population in the Tuscan
Seas. ARPAT - Settore Mare – Area vasta Costa, Firenze (49 pp.). http://www.arpat.
toscana.it/documentazione/report/.
Allen, H.E., Fu, G., Deng, B., 1993. Analysis of acid-volatile sulfide (AVS) and
simultaneously extracted metals (SEM) for the estimation of potential toxicity in
aquatic sediments. Environ. Toxicol. Chem. 12, 1441–1453.
ARPAT, 2016. Monitoring on the "Management of the Lagoon of Orbetello through
sediment handling induced by boats transit. Report for Tuscany Region, archiving
protocol 0071294, 28/10/2016. http://www.arpat.toscana.it/.
Ausili, A., Berducci, M.T., Maggi, C., Sesta, G., 2018. Analisi di sostanze prioritarie in
matrici marine. Parte I. Verifica delle metodologie ufficiali esistenti e loro
applicabilità alle matrici marine. ISPRA – Manuali e Linee Guida 175/2018, 44pp.
Roma febbraio 2018. https://www.isprambiente.gov.it/files2018/pubblicazioni/ma
nuali-linee-guida/MLG_175_18.pdf.
Beccaloni, M.R., Cicero, A.M., Mancini, L., Marcheggiani, S., Miniero, R., Scenati, R.,
Vendetti, C., Ziemacki, G., Carere, M., 2011. Approaches for the derivation of
sediment quality criteria for the protection of human health in Italian water bodies.
In: Proceedings Sixth International Conference on Remediation of Contaminated
Sediments (New Orleans, Louisiana; February 7–10, 2011). Institutions, Report for.
https://www.minambiente.it/sites/default/files/bonifiche/contenuti/Orbetell
o/2013/c_istruttorie/verbale%2029.5.13.pdf.
Benoit, J.M., Gilmour, C.C., Mason, R.P., Heyes, A., 1999. Sulfide controls on mercury
speciation and bioavailability to methylating bacteria in sediment pore waters.
Environmental Science & Technology 33 (10), 1780. https://doi.org/10.1021/
es9808200.
Benoit J.M., Gilmour C.C., Heyes A., Mason R.P., Miller C.L., 2003. Geochemical and
biological controls over methylmercury production and degradation in aquatic
ecosystems, Biogeochemistry of Environmentally Important Trace Elements. Acs
Symposium Series, 262–297.
Benoit, J.M., Shull, D.H., Robinson, P., Ucran, L.R., 2006. Infaunal burrow densities and
sediment monomethyl mercury distributions in Boston Harbor. Massachusetts. Mar.
Chem. 102 (1), 124–133.
Boening, D.W., 2000. Ecological effects, transport, and fate of mercury: a general review.
Chemosphere 40, 1335–1351. https://doi.org/10.1016/S0045-6535(99)00283-0.
Caetano, M., Madureira, M.-J., Vale, C., 2002. Metal remobilisation during resuspension
of anoxic contaminated sediment: short-term laboratory study. Water Air Soil
Pollution 143 (1–4), 23–40.
Caille, N., Tiffreau, C., Leyval, C., Morel, J.L., 2003. Solubility of metals in an anoxic
sediment during prolonged aeration. Sci. Total Environ. 301, 239–250.
Castoldi, A.F., Onishchenko, N., Coccini, T., Roda, E., Vahter, M., Ceccatelli, S.,
Manzo, L., 2008. Human developmental neurotoxicity of methylmercury: impact of
variables and risk modifiers. Regul. Toxicol. Pharmacol. 51 (2), 201–214. https://
doi.org/10.1016/j.yrtph.2008.01.016.
Chakraborty, P., Manek, A., Niyogi, S., Hudson, J., 2014. Determination of dynamic
metal complexes and their diffusion coefficients in the presence of different humic
substances by combining two analytical techniques. Anal. Lett. 47, 1224–1241.
11
Marine Pollution Bulletin 162 (2021) 111907
Chen, C.Y., Serrell, N., Evers, D.C., Fleishman, B.J., Lambert, K.F., Weiss, J., Mason, R.P.,
Bank M.S., 2008. Meeting report: methylmercury in marine ecosystems-from sources
to seafood consumers. Environ. Health Perspect. 116, 1706–1712.
Chen, C.Y., Dionne, M., Mayes, B.M., Ward, D.M., Sturup, S., Jackson, B.P., 2009.
Mercury bioavailability and bioaccumulation in estuarine food webs in the Gulf of
Maine. Environ. Sci. Technol. 43, 1804–1810.
Clarisse, O., Lotufo, G.R., Hintelmann, H., Best, E.P.H., 2012. Biomonitoring and
assessment of monomethylmercury exposure in aqueous systems using the DGT
technique. Sci. Total Environ. 416, 449–454.
Coelho, J.P., Pereira, M.E., Duarte, A.C., Pardal, M.A., 2009. Contribution of primary
producers to mercury trophic transfer in estuarine ecosystems: possible effects of
eutrophication. Mar. Pollut. Bull. 58, 358–365.
Compeau, G.C., Bartha, R., 1985. Sulfate-reducing bacteria: principal methylators of
mercury in anoxic estuarine sediment. Applied and Environtal Microbiology 50,
498–502.
Corder, G.W., Foreman, D.I., 2014. Nonparametric statistics: a step-by-step approach.
Wiley. In: Hollander, M., Wolfe, D.A., Chicken, E. (Eds.), Nonparametric Statistical
Methods. John Wiley & Sons.
Dash, S., Das, S.K., Samal, J., Thatoi, H.N., 2018. Epidermal mucus, a major determinant
in fish health: a review. Iranian Journal of Veterinary Research, Shiraz University 19
(2, Ser. 63), 72–81.
Davison, W., Zhang, H., 1994. In situ speciation measurements of trace components in
natural waters using thin film gels. Nature 367, 546–548.
Di Toro, D.M., Mahony, J.D., Hansen, D.J., Scott, K.J., Hicks, M.B., Mayr, S.M., et al.,
1990. Toxicity of cadmium in sediments: the role of acid volatile sulfide. Environ.
Toxicol. Chem. 9, 1487–1502.
Díez, S., Giaggio, R., 2018. Do biofilms affect the measurement of mercury by the DGT
technique? Microcosm and field tests to prevent biofilm growth. Chemosphere 210,
692–698.
Fernández-Gómez, C., Dimock, B., Hintelmann, H., Díez, S., 2011. Development of the
DGT technique for Hg measurement in water: comparison of three different types of
samplers in laboratory assays. Chemosphere 85, 1452–1457.
Fleming, E.J., Mack, E.E., Green, P.G., Nelson, D.C., 2006. Mercury methylation from
unexpected sources: molybdate-inhibited freshwater sediments and an iron-reducing
bacterium. Applied Environmental Microbiology 72 (1), 457–464.
Froese, R., Pauly, D. (Eds.), 2004. FishBase. World Wide Web electronic publication.
http://www.fishbase.org/. version 11.
Gill, G.A., Bloom, N.S., Cappellino, S., Driscoll, C.T., Dobbs, C., McShea, L., Mason, R.P.,
Rudd, G.W.M., 1999. Sediment-water fluxes of mercury in Lavaca Bay, Texas.
Environmental Science & Technology 33 (5), 663–669. https://doi.org/10.1021/
es980380c.
Harris, R.C., Krabbenhoft, D.P., Mason, R., Murray, M.W., Reash, R., Saltman, T. (Eds.),
2007. Ecosystem Responses to Mercury Contamination: Indicators of Change. CRC
Press, Boca Raton, FL, USA.
Hollander, M., Wolfe, D.A., Chicken, E., 2014. Nonparametric Statistical Methods. John
Wiley & Sons.
Holm, S., 1979. A simple sequentially rejective multiple test procedure. Scand. J. Stat. 6
(2), 65–70.
Horvat, M., 1977. Mercury behavior in estuarine and coastal environment. In: In: Rajar,
R., Brebbia, C.A. (Eds.), Water Pollution IV. Modelling, Measuring and Prediction
Proceedings of the 4th International Conference on Water Pollution, Lake Bled,
Slovenia. Computational Mechanics Publications, Southampton, UK, pp. 77–86.
Hurley, J., Benoit, J., Babiarz, C., Shafer, M., Andren, A., Sullivan, J., Hammond, R.,
Webb, D., 1995. Influence of watershed characteristics on mercury levels in
Wisconsin rivers. Environ. Sci. Technol. 29, 1867–1875.
ICRAM – UniSi, Central Institute for Marine Research and Siena University, 2008.
Relazione Finale. Piano di Indagine Finalizzato alla Bonifica ed al Risanamento
Ambientale della Laguna di Orbetello. 1◦ Stralcio attuativo (Report for Institutions).
Jørgensen B.B., 1983. The microbial sulphur cycle. In: Krumbein, W. (Ed.), Microbial
Geochemistry. Blackwell Scientific Publications, Oxford, pp. 91–124.
Kannan, K., Smith Jr., R.G., Lee, R.F., Windom, H.L., Heitmuller, P.T., Macauley, J.M.,
Summers, J.K., 1998. Distribution of total mercury and methyl mercury in water,
sediment, and fish from South Florida Estuaries. Arch. Environ. Contam. Toxicol. 34,
109–118.
King, J.K., Kostka, J.E., Frischer, M.E., Saunders, F.M., Jahnke, R.A., 2001. A quantitative
relationship that demonstrates mercury methylation rates in marine sediments are
based on the community composition and activity of sulfate-reducing bacteria.
Environ. Sci. Technol. 35, 2491–2496. https://doi.org/10.1021/es001813q.
Lawrence, A.L., McAlloon, K.M., Mason, R., Mayer, L.M., 1999. Intestinal solubilization
of particle-associated organic and inorganic mercury as a measure of bioavailability
to benthic invertebrates. Environmental Science & Technology 33 (11), 1871–1876.
Lenzi M., 2010. Resuspension of sediment as method for managing shallow eutrophic
lagoon. Journal of Ecology and the Natural Environment 2 (11), 220-234. (ISSN
2006-9847). http://academicjournals.org/jene.
Lenzi, M., Palmieri, R., Porrello, S., 2003. Restoration of the eutrophic Orbetello lagoon
(Tyrrhenian Sea, Italy): water quality management. Mar. Pollut. Bull. 46,
1540–1548.
Lenzi, M., Birardi, F., Calzolai, R., Finoia, M.G., Marcone, F., Nocciolini, S., Roffilli, R.,
Sgroi, S., Solari, D., 2010. Hypertrophic lagoon management by sediment
disturbance. Mar. Pollut. Bull. 61 (4–6), 189–197.
Lenzi, M., Renzi, M., Nesti, U., Gennaro, P., Persia, E., Porrello, S., 2011. Vegetation
cyclic shift in eutrophic lagoon. Assessment of dystrophic risk indices based on
standing crop evaluation. Estuarine. Coastal and Shelf Science 132, 99–107. https://
doi.org/10.1016/j.ecss.2011.10.006.
M. Lenzi et al.
Lenzi, M., Finoia, M.G., Gennaro, P., Mercatali, I., Persia, E., Solari, J., Porrello, S., 2013.
Assessment of resuspended matter and redistribution of macronutrient elements
produced by boat disturbance in a eutrophic lagoon. J. Environ. Manag. 123, 8–13.
Lenzi, M., Salvaterra, G., Gennaro, P., Mercatali, I., Persia, M., Porrello, S., Sorce, C.,
2015. A new approach to macroalgal bloom control in eutrophic, shallow water,
coastal areas. J. Environ. Manag. 150, 456–465. https://doi.org/10.1016/j.
jenvman.2014.12.031.
Lenzi, M., Persiano, M., Gennaro, P., Rubegni, F., 2016. Wind mitigating action on effects
of eutrophication in coastal eutrophic water bodies. International Journal of Marine
Science and Ocean Technology 3 (2), 14–20.
Lenzi, M., Persiano, M., Gennaro, P., Rubegni, F., 2017. Artificial top layer sediment
resuspension to counteract Chaetomorpha linum (Muller) Kutz. Blooms in a
eutrophic lagoon. Three years full-scale experience. Journal Aquaculture Marine
Biology 5 (2), 00114. https://doi.org/10.15406/jamb.2017.05.00114.
Lenzi, M., Gennaro, P., Franchi, E., Marsili, L., 2019. Assessment of fish-farms
wastewater synergistic impact on a Mediterranean non-tidal lagoon. Journal of
Aquaculture and Fisheries 3, 021. https://doi.org/10.24966/AAF-5523/100021.
Martelloni, T., Tomassetti, P., Gennaro, P., Vani, D., Persia, E., Persiano, M., Falchi, R.,
Porrello, S., Lenzi, M., 2016. Artificial soft sediment resuspension and high density
opportunistic macroalgal mat fragmentation as method for increasing sediment
zoobenthic assemblage diversity in a eutrophic lagoon. Mar. Pollut. Bull. 110 (1),
212–220. https://doi.org/10.1016/j.marpolbul.2016.06.060.
Mason, R.P., 2002. The bioaccumulation of mercury, methylmercury, and other toxic
elements into pelagic and benthic organisms. In: Newman, M.C., Roberts, M.H.,
Hale, R.C. (Eds.), Coastal and Estuarine Risk Assessment. CRC Press, Boca Raton, FL,
pp. 127–149.
Mason, R., Kim, E.H., Cornwell, J., Heyes, D., 2006. An examination of the factors
influencing the flux of mercury, methylmercury and other constituents from
estuarine sediment. Mar. Chem. 102, 96–110. https://doi.org/10.1016/j.
marchem.2005.09.021.
McAlloon, K.M., Mason, R., 2003. Investigations into the bioavailability and
bioaccumulation of mercury and other trace metals to the sea cucumber,
Sclerodactyla briareus, using in vitro solubilization. Mar. Pollut. Bull. 46, 1600–1608.
12
Marine Pollution Bulletin 162 (2021) 111907
McGlathery, K., Sundbäck, K., Anderson, I.C., 2007. Eutrophication in shallow coastal
bays and lagoons: the role of plants in the coastal filter. Mar. Ecol. Prog. Ser. 348,
1–18.
Miniero, R., Beccaloni, E., Carere, M., Ubaldi, A., Mancini, L., Marchegiani, S., Cicero, M.
R., Scenati, R., Lucchetti, D., Ziemacki, G., De Felip, E., 2013. Mercury (Hg) and
methyl mercury (MeHg) concentrations in fish from the coastal lagoon of Orbetello,
central Italy. Mar. Pollut. Bull. 76 (1–2), 365–369.
Pan, J.F., Wang, W.-X., 2004. Uptake of Hg(II) and methylmercury by the green mussel
Perna viridis under different organic carbon conditions. Mar. Ecol. Prog. Ser. 276,
125–136.
Peat J, Barton B., 2005. A Guide to Data Analysis and Critical Appraisal. Blackwell
Publishing. Medical Statistics. ISBN-10: 9780727918123.
Rubegni, F., Franchi, E., Lenzi, M., 2013. Relationship between wind and seagrass
meadows in a non-tidal eutrophic lagoon studied by a wave exposure model
(WEMo). Mar. Pollut. Bull. 70 (1–2), 54–63. https://doi.org/10.1016/j.
marpolbul.2013.02.012.
Saulnier, I., Mucci, A., 2000. Trace metal remobilization following the resuspension of
estuarine sediments: Saguenay Fjord, Canada. Appl. Geochem. 15, 191–210.
Simpson, S., Capte, S., Batley, G., 1998. Effect of short-term resuspension events on trace
metal speciation in polluted anoxic sediments. Environmental Science & Technology
32, 620–625.
StataCorp, 2015. Stata: Release 14. Statistical software. College Station, StataCorp LP.
Tremolada, P., 1998. Cicli Biogeochimici. In: In —Ecotossicologia‖. UTET, Volume a
cura di M. Vighi e E. Bacci.
Wiener, J.G.K., Krabbenhoft, D.P., Heinz, G.H., Scheuhammer, A.M., 2003.
Ecotoxicology of mercury. In: Burton Jr., G.A., Cairns Jr., J. (Eds.), Hoffman, D.J.R.,
B.A. CRC Press, Boca Raton, FL, USA, Handbook of Ecotoxicology, pp. 409–443.
Wu, T., Wang, G., Zhang, Y., Kong, M., Zhao, H., 2017. Determination of mercury in
aquatic systems by DGT device using thiol-modified carbon nanoparticle suspension
as the liquid binding phase. New J. Chem. 41 (18), 10305–10311.