Marine Pollution Bulletin 131 (2018) 248–251

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Marine Pollution Bulletin

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

Note

Microplastic contents from maricultured and natural mussels T

⁎
Monia Renzi , Cristiana Guerranti, Andrea Blašković
Bioscience Research Center, via Aurelia Vecchia, 32, 58015 Orbetello, Italy

A R T I C LE I N FO A B S T R A C T

Keywords: Results of this research focuses on microplastic contents (levels, type, size, colour) in maricultured and natural
Marine trophic web mussels (Mytilus galloprovincialis) from different Italian stocks. No significant differences were found among
Diet intake maricultured and natural stocks. All recovered MPs are filaments ranging within 750–6000 μm of maximum
Human risks length (average values 1150–2290 μm). Feeding raw mussel could produce median MP intakes of 6.2–7.2 items/
Microplastics
g w.w. Concerning human exposure by diet, both raw and cooked values are important. Some preliminary tests
Mussel
performed in this study evidenced that the cooking process determined lower MPs levels (−14%) in cooked
Mariculture
tissues compared to raw ones, MPs were recorded in cooking water and were characterized by a lower size than
in raw mussels. Results obtained by this study represent an important baseline on MPs level to evaluate en-
vironmental and human exposure risks by diet.

There is a global concern over the impact of plastic waste, mainly microplastics including effects of food processing (EFSA, 2016).
microplastics (MPs, particles of size below 5 mm, according to NOAA, Given the above needs on data collection and in response to the
2008) found in seas and rivers, on natural habitats and wildlife. MPs indications of EFSA, this study was performed in order to attain the
could represent significant risks for marine habitats and could affect following purposes: i) defining levels and principal features (type, size,
marine species and top predators throughout marine trophic webs. colour) of MPs in mussels (Mytilus galloprovincialis, Lamarck 1819) from
Furthermore, a potential concern is related to the high concentrations maricultured and natural stocks; ii) comparing MPs levels and principal
of contaminants such as persistent organic pollutants (POPs), which can features measured in maricultured from various Italian plants to levels
be absorbed by MPs in the environment (Cole et al., 2011) or may also in natural stocks; iii) evaluating differences occurring among MPs levels
be present in plastic material since used in packaging, such as bisphenol in raw and cooked mussels.
A (BPA) or phthalates (Fossi et al., 2016). At the best of our knowledge, Three different commercial stocks of M. galloprovincialis coming
there are not studies highlighting as chemicals from microplastics can from different Italian mariculture plants (Cesenatico, central Adriatic
be transferred to organisms along the trophic web and these aspects Coast; La Spezia, Ligurian Sea Coast, and Olbia, North East Sardinia,
should be improved by further researches. Intake of MPs by mussels has Tyrrhenian Sea) were collected from local markets. A natural stock of
been reported by the literature (Browne et al., 2008; Santana et al., the same species was sampled from natural rocky bottom in Tuscany
2016). Mussels are interesting species to evaluate risks associated to (Talamone, central Tyrrhenian Sea) to evaluate differences among
MPs in marine habitats due to they can: -) accumulate MPs in water commercial and natural stocks concerning MP levels and principal
column by filtering; −) transfer MPs from water column towards the features. Collected animals were measured to determine biometrics
marine trophic web as they are predated by many marine species (i.e. (maximum length, mm; weight, g) to build for each stock the size curve
carnivorous gastropods, sea breams, starfish); −) affect human health of the sampled population. A representative number of animals were
by diet based on raw or cooked mussels. In spite of mussels importance collected from each tested stock selecting them from the same size class
in MPs transfer from water column through the trophic web towards and preferring more represented size classes. Animals were analysed
humans, no data are reported on MPs levels in natural mussels stocks individually rather than in pools to improve data representativeness
and commercial products to evaluate both ecosystem and humans and statistic significant of collected data. Raw animals from the same
health risks associated to mussel consumption. In particular, concerning size class (n = 10) were dissected to open valves, hepatopancreas and
humans, it is important to estimate the average intake; EFSA estimated gills were excised thanking care not to damage them, weighted and
the amount of MPs contained in a portion of mussels and, at the same extracted. To evaluate the effect of cooking, another group of animals
time, stated that research should produce data on the presence of from the same stock (n = 10) were cooked independently in an

⁎
Corresponding author.
E-mail address: [email protected] (M. Renzi).

https://doi.org/10.1016/j.marpolbul.2018.04.035
Received 19 March 2018; Received in revised form 11 April 2018; Accepted 12 April 2018
0025-326X/ © 2018 Published by Elsevier Ltd.
M. Renzi et al. Marine Pollution Bulletin 131 (2018) 248–251

accurately rinsed glass backer with 50 mL of deionized water. Cooking MPs recovered per animals in each tested stock are reported. Cesenatico
time was standardized in 2 min of cooking in boiling water, to simulate stock showed highest items values per animals recorded. Significant
a real cooking process. Then cooked mussels and cooking water were differences are recorded between the following couples: Cesenatico-La
analysed separately. In particular, cooked tissues were excised to re- Spezia (p = 0.002), Cesenatico-Olbia (p < 0.001), Cesenatico-Tala-
move hepatopancreas and gills, weighted and extracted to recover mone (p < 0.001). Low significant is reported for the couple Tala-
litter. Extraction was performed in glass backer with 20 mL of H2O2 mone-La Spezia (p = 0.027). To standardize collected data, in Fig. 2
30% per gram of tissue, backers were put in a Bain-marie at 50 °C for (red bars), average (standard deviation) items of MPs recovered per
48 h (raw tissues) but, if needed, the extraction time was extended till gram of tissue in each tested stock are reported. Cesenatico stock
the complete digestion of tissues (cooked tissue) as adapted by Nuelle showed the lower variability among specimens on the contrary, the
et al. (2014). Extracted sample (raw and cooked tissues) were com- stock from La Spezia showed the highest variability recorded. Any
pletely recovered onto paper fibres (0.45 μm filter disks) by the use of a difference was recorded between Olbia and Talamone stocks. Except for
filtrating system, consisting in filtration glass set, vacuum pump, the couple Cesenatico-Olbia (p < 0.05), statistics evidence no sig-
manifold and funnel. Glassware was accurately rinsed to increase re- nificant differences among average values concerning tested stocks. In
covery efficiency of litter. Filters were stored in glass Petri dish and the scientific literature, few studies focus on the measurements of the
dried in oven at 40 °C till constant weight. Cooking water was filtered concentrations of MP in molluscs both present in the environment and
directly taking care to recover by rinsing any plastic items from the maricultured. Van Cauwenberghe and Janssen (2014), for example,
backer walls and filters were dried as reported. The plastic items were reported, for Mytilus edulis, therefore comparable with the organism
determined by stereomicroscopy (Nikon SMZ-800 N) and all identified investigated in this study (never studied before for what concerns the
items were divided in shape, colour and dimensional classes reported presence of MPs, for the authors knowledge), average levels of 0.36 MP
by the literature (Galgani et al., 2013; Alomar et al., 2016; Blaskovic items/g w.w. Therefore the average value found in this study is, al-
et al., 2017; Fastelli et al., 2016). The Nikon's software for the imaging though for a different species from that of comparison, much higher
analysis was applied to the litter dimensional measurements (Nikon than reported by the above authors. Further researches are needed to
ACT-1). Data on litter items counting and litter dimensions are reported clarify the reason of observed data even if some hypotheses are related
as mean + standard deviation or as median. Population size–class to MPs load differences among geographical areas of origin of the
curves were determined by Excel® or Prism® routines. Statistics were analysed molluscs, or to differences among different stocks MPs accu-
performed for data analyses (Prism software, Graph-pad Software) mulation/excretion, which must be further investigated. MPs transfer
considering a p < 0.01 statistically significant. All the filters were through the food web has been demonstrated for mussel (Farrell and
checked by the four-eye approach and by an inter-calibration process Nelson, 2013), with important implications for the health of marine
performed between two operators; in order to minimise accidental organisms, food chains, and for human health. Commercial and natural
contaminations, only glass materials and cotton dresses were used by stocks are not significantly different concerning MP contents in raw
operators and to treat samples and tests were performed on blanks. animals with the exception of Cesenatico and Olbia. This difference
Filters (n = 3) were left overnight exposed to the laboratory air, putting could be probably due to different MPs pollution levels reported by the
them on the desk on an opened glass Petri dish and analysed to check literature between the Adriatic Sea and the Tyrrhenian Sea (Blašković
air laboratory blanks. Other filters were extracted as reported for raw et al., 2017; Guerranti et al., 2017; Cannas et al., 2017). Colour dom-
tissue samples; cooking blanks were, also, performed and checked by inances are similar to dominances recorded by the above-mentioned
the four-eye approach to detect MPs. All blanks analysed resulted free literature in sediments. Black and Blue are the most represented colours
from MPs. in all tested stocks. Red and Pink show low frequencies, while Green,
In Fig. 1 the population structure of tested commercial and natural Orange, Yellow, Grey and Brown coloured microplastics were only
stocks is reported. Tested stocks ranged within 30–90 mm valve length occasionally recorded. All recovered MPs are filaments ranging within
with some differences. Stocks coming from La Spezia and Olbia are 0.75–6.00 mm of length even if average values are included within
similar and range respectively within 40–75 mm (mode 50–55 mm) and 1.15–2.29 mm (Table 1). The fact that filaments are the only MPs type
40–85 mm (mode 60–65 mm). Specimens from Cesenatico are larger recorded in tested stocks could probably be due to the geometry of
and it ranges within 60–90 mm (mode 65–70 mm) while these coming these plastics that allow them to better reach the considered species.
from Talamone are smaller and ranges within 30–65 mm (mode Furthermore, filaments get trapped into gills and hepatopancreas and
45–50 mm). In Fig. 2 (blue bars), average (standard deviation) items of cannot be easily removed by animals accumulating into them.

Fig. 1. Population structure of tested stocks. The number of individuals per each dimension class (5 mm length) are represented in tested stocks.

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M. Renzi et al. Marine Pollution Bulletin 131 (2018) 248–251

Fig. 2. Average values (SD) of microplastics recovered in raw animals from different commercial and natural stocks. Data are reported both as average items per
animal and as average items per gram of tissue.

Table 1
Microplastic items mean length and standard deviation (SD) in tested mussels
stocks.
Stock Mean length (mm) SD

Raw Cesenatico 1.70 0.37

La Spezia 1.90 0.75
Olbia 1.71 0.52
Talamone 1.89 0.57
Cooked Cesenatico 1.40 0.56
La Spezia 2.04 0.80
Olbia 1.65 0.91
Talamone 2.27 1.14
Cooking water Cesenatico 1.00 0.11
La Spezia 1.15 0.34
Olbia 2.29 0.71
Talamone 1.83 0.17

Concerning human exposure to MPs ingestion by diet and associated

health risks, the European Food Safety Authority (EFSA) has recently
examined the existing literature on the subject (EFSA, 2016, noting the
inadequacy of available data on the presence, toxicity and fate, what
happens for instance during and after the digestion of such materials,
aiming a complete risk assessment. In Fig. 3a, average (+SD) value of
recovered number of items per gram of tissue is reported for cooked
tissues and cooking water as percentage compared to levels in raw
tissues. Median levels are included within 6.2–7.2 items/g of tissue.
Cooked tissues show on average lower MPs levels than raw ones. This
data is associated to the presence of litter in cooking water. It should be
noted (Table 1) that the length of MPs measured in raw tissues
(1.80 mm) is similar to values recorded in cooked tissues (1.84 mm)
even if standard deviations differ significantly (0.11 mm vs. 0.39 mm
respectively). On the contrary, average value recorded in cooking water Fig. 3. a) Average (SD) values of microplastics recovered in cooked tissues and
is lower and associated to a higher variability (1.76 mm; cooking water. Data are reported as percentage of number of items per gram of
tissue w.w. compared to raw samples. b) Average values of maximum length
SD = 0.57 mm). In the cooking water, we find relevant level of MPs
(mm + SD) of microplastics collected in raw, cooked tissues and cooking water.
expressed as number of items/g tissue, which added to the values,
found in cooked organisms, they give a total levels higher than founded
in raw mussels. The observed results could be probably due to two better clarify this aspect. High plastic debris concentrations have been
factors able to determine observations performed: i) natural variability found in fish, but since the MPs are present mostly in the stomach and
of items inside individuals from the same stock; ii) thermal degradation intestine, which are usually removed before the human consumption,
of microplastics during cooking which could produce a higher number resulting in an absence of exposure for consumers. However, in the case
of items/g in cooking water and cooked tissues associated to a lower of crustaceans and bivalve mollusks, such as oysters and mussels, the
size as reported in Fig. 3b (average, mm; +SD). Temperature closed to digestive tract is eaten, leading to some exposure (EFSA, 2016). In bi-
water boiling point, in fact, could determine micro fractures or fusion of valves in general, the average number of items of MPs is 0.2–4.0 items/
the polymeric structure of some plastic materials (i.e. fusion tempera- g (EFSA, 2016), even if extraction methods are different, values re-
ture for PVC 100–260 °C; PE 115–140 °C) and can probably induce ported by the literature are lower if compared to the average value
fragmentation. Nevertheless, further studies could be performed to measured in this study. EFSA (2016), also, estimated that the

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