Journal of

Marine Science
and Engineering

Article
Bioremediation Capabilities of Hymeniacidon perlevis
(Porifera, Demospongiae) in a Land-Based Experimental
Fish Farm
Caterina Longo 1 , Cataldo Pierri 1 , Maria Mercurio 1 , Roberta Trani 1, * , Frine Cardone 2 ,
Pierluigi Carbonara 3 , Sébastien Alfonso 3 and Loredana Stabili 4,5

1 Department of Biology, University of Bari Aldo Moro, Via Orabona 4, 70125 Bari, Italy;
[email protected] (C.L.); [email protected] (C.P.); [email protected] (M.M.)
2 Department of Integrative Marine Ecology, Zoological Station “Anton Dohrn”, Villa Comunale,
80121 Naples, Italy; [email protected]
3 COISPA Tecnologia & Ricerca, Stazione Sperimentale per lo Studio delle Risorse del Mare,
Via dei Trulli 18/20, Torre a Mare, 70126 Bari, Italy; [email protected] (P.C.); [email protected] (S.A.)
4 Water Research Institute (IRSA)-Consiglio Nazionale delle Ricerche (CNR), 74123 Taranto, Italy;
[email protected]
5 Department of Biological and Environmental Sciences and Technologies, University of Salento,
73047 Lecce, Italy
* Correspondence: [email protected]

Abstract: The expansion of aquaculture practices in coastal areas can alter the balance of microbial
communities in nearby marine ecosystems with negative impacts on both farmed and natural species,
as well as on human health through their consumption. Among marine filter-feeder invertebrates,
poriferans are known as effective microbial bioremediators, even though they are currently still
underutilized in association with fish mariculture plants. In this study, we investigate the microbial
Citation: Longo, C.; Pierri, C.;
bioremediation capability of the demosponge Hymeniacidon perlevis in an experimental land-based
Mercurio, M.; Trani, R.; Cardone, F.;
fish farm where this species occurred consistently in the drainage conduit of the wastewater. Microbi-
Carbonara, P.; Alfonso, S.; Stabili, L.
Bioremediation Capabilities of
ological analyses of cultivable vibrios, total culturable bacteria (37 ◦ C), fecal and total coliforms, and
Hymeniacidon perlevis (Porifera, fecal enterococci were carried out on the fish farm wastewater in two sampling periods: autumn and
Demospongiae) in a Land-Based spring. The results showed that H. perlevis is able to filter and remove all the considered bacterial
Experimental Fish Farm. J. Mar. Sci. groups from the wastewater, including human potential pathogens, in both sampling periods. This
Eng. 2022, 10, 874. https://doi.org/ finding sustains the hypothesis of H. perlevis use as a bioremediator in land-based aquaculture plants
10.3390/jmse10070874 as well.
Academic Editor: Azizur Rahman
Keywords: bacterial bioremediation; Hymeniacidon perlevis; land-based fish farm
Received: 6 June 2022
Accepted: 23 June 2022
Published: 25 June 2022

Publisher’s Note: MDPI stays neutral 1. Introduction

with regard to jurisdictional claims in Porifera, or sponges, are the oldest metazoans that are exclusively aquatic, mainly
published maps and institutional affil- marine, that are still extant. They are primitive pluricellular animals whose functions are
iations. based on a flexible differentiation system characterized by a high plasticity at the cellular,
structural, and reproductive levels [1,2]. These features make them able to adopt a wide
range of adaptive strategies for population survival and support great success in benthic
communities. Sponges are effective filter-feeders that are able to process a huge water
Copyright: © 2022 by the authors.
volume and retain up to 99% of the particulate and dissolved organic matter [3–5]. These
Licensee MDPI, Basel, Switzerland.
sessile metazoans combine sexual and asexual reproduction [6]. This latter reproductive
This article is an open access article
strategy, carried out by means of fragmentation, budding, and gemmulae, allows for the
distributed under the terms and
conditions of the Creative Commons
spread of numerous well-adapted individuals with a higher chance of survival. Their
Attribution (CC BY) license (https://
cellular and reproductive plasticity give them a characteristic regenerative ability that
creativecommons.org/licenses/by/ allows for the healing of damaged or lost parts, as well as the complete regeneration of an
4.0/). adult from fragments or even single cells.

J. Mar. Sci. Eng. 2022, 10, 874. https://doi.org/10.3390/jmse10070874 https://www.mdpi.com/journal/jmse

J. Mar. Sci. Eng. 2022, 10, 874 2 of 13

The ability of sponges to grow from fragments excised from wild donor sponges, and
then seed a range of standard aquaculture infrastructure, makes them ideal candidates for
aquaculture [7–10].
Recently, in order to develop methods for “artificial seed” production and sustainable
supply in sponge aquaculture, an intensive study on Hymeniacidon perlevis (Montagu, 1818)
larval release and settlement under controlled conditions has been carried out [11–13].
Nevertheless, harvesting from wild donor stock and a subsequent culture of explants is
currently the most feasible method to obtain valuable quantities of sponge [14].
Several sponge species show commercial potential use, including as biomaterials. In
this framework, there is increased interest in investigating sponge features, not only for
their biological relevance per se but also for the contributing information that can be used
in the context of applied research [15].
Bacteria are important components of sponge holobionts (Ref. [16] and references
therein) and shifts in the composition of the microbiota can compromise the health of the
whole animal [17,18]. The maintenance of a characteristic microbial community appears to
be a complex trait, suggesting that hosts deprived by their normal microbiota should be at
a disadvantage [2].
The excessive release of microbial pathogens, such as bacteria, viruses, and protozoans
that come from human and animal waste, has become a major concern with increasing
coastal urbanization and aquaculture practices because of their potential impact on coastal
marine environments [19,20]. First, pathogenic bacteria represent a significant health hazard
to aquatic species and can cause recurrent occurrences of marine diseases. Second, seafood
products are severely contaminated by pathogenic bacteria and represent a biohazard to
human health through human consumption [21].
In this scenario, scientists all over the world are working towards the development of
remediation technologies, including physical, chemical, and biological remediation (named
bioremediation). Bioremediation is a potential technique for the biological treatment of
industrial waste and contaminated environments [22–24]. Recent research has shown that,
apart from the utilization of microorganisms for the biodegradation of target pollutants,
aquatic ecosystems are home to several invertebrate species who deserve the definition of
zooremediators [25] due to their ability to remove, hyperaccumulate, stabilize, or degrade
pollutants. Among them, marine sponges have demonstrated the capability to remediate
aquatic microbial pollution [26–36] and accumulate or degrade metals [37–43]. In addition,
sponges host massive consortia of microorganisms within the mesohyl matrix that can
amount for up to 60% of their total biomass [44], exceeding that of seawater by two to three
orders of magnitude [16] and significantly contributing to the host metabolism [45].
Among poriferans, H. perlevis has been shown to be an effective bioremediator that
is able to remove potential pathogenic bacteria from the cultured media by retaining and
digesting them by phagocytosis [26,27,31,32,34,46]. In addition, recent research [40] has
shown that H. perlevis has impressive bioaccumulation properties for different organic
and inorganic pollutants (particularly metallic trace elements and polycyclic aromatic
hydrocarbons). Moreover, their associated microbiota have been demonstrated to have
capability in lindane pesticide degradation [47,48]. These features make H. perlevis suitable
for the bioremediation of polluted seawater by microbial, organic, and inorganic pollutants,
such as in aquaculture farming plants.
Most of the known studies are detailed laboratory-based studies that were not performed
in situ in a true aquaculture system, except those of Longo et al. [32] and Zhang et al. [27].
The occurrence of large amounts of the demosponge H. perlevis consistently living
in the drainage conduit of the wastewater flowing out from an experimental land-based
fish farm offered the opportunity to investigate its ability for microbial reduction in such
conditions, and to evaluate the possibility to use this species in integrated multitrophic
farming systems for wastewater treatment, which is in contrast to traditional technologies
with high negative impacts on the whole ecosystem.
J. Mar. Sci. Eng. 2022, 10, 874 3 of 13

2. Materials and Methods

2.1. Studied Species
The Demospongiae, Hymeniacidon perlevis, family Halichondriidae Gray, 1867, is ex-
tremely plastic in shape and can form encrusting or massive specimens that are able to
survive prolonged periods of air exposure [49,50]. This sponge is typically characterized
by orange to red or pale green specimens that can reach up to 30 cm in diameter. Its surface
is characterized by emerging short papillae and small digitations.
H. perlevis is one of the most common species of Europe [51], it has a wide distribution,
is considered cosmopolitan [52], and currently is the most common and widely-distributed
exotic sponge [53].
H. perlevis, predominantly reported from temperate and colder waters, lives in shal-
low subtidal and intertidal zones where it can colonize both hard substrates and soft
bottoms, and where it is able to live buried into the sediment with its oscules projecting
outwards [51,54]. This species is one of the most common in the central Mediterranean
lagoon systems and has recently been recorded in a Turkish lagoon [50,55], where it inhabits
an environment deeply affected by anthropogenic impact. H. perlevis is able to survive both
air exposure and strong environmental parameter changes [49,55]. Despite local seasonal
variations in growth and population size [49,56,57], this species shows a peculiar adaptive
strategy, thereby allowing it to face critical environmental conditions.

2.2. In-Situ Bioremediation Study

Bioremediation studies were performed at the COISPA, a land-based experimental fish
farm station (Torre a Mare, southern Italy; www.coispa.it (accessed on 23 June 2022)) where
Hymeniacidon perlevis lives on the bottom of the drainage conduit for wastewater (Figure 1).
This conduit receives wastewater from the fish farming tanks and conveys it into the sea.
The drainage conduit considered in this study consists of a cement canal that is 4.0 m long,
0.57 m wide, and 0.15 m deep, and is closed by a walkable inspection protection that makes
this part of the conduit completely dark (Figure 1). The water used in this land-based fish
farm is sea-groundwater and its temperature is constant throughout the year (18 ◦ C). The
wastewater from the fish tanks flows out through the drainage conduit and passes through
the area with sponges. Each tank has an overflow system and also a tap at the bottom.
To eliminate feed and feces residues that accumulate at the bottom of the tank, the tap of
each tank is opened daily and approximately one-third of the entire volume of the tank is
discharged. The bioremediation experiment was performed during two sampling periods:
spring and autumn. The fish biomass in the system was similar in both sampling periods,
ranging between 590 and 660 kg in spring and autumn, respectively. The water volume
of the fish tanks was 27,990 and 35,190 L in spring and autumn, respectively. The volume
of the sponges living inside the drainage conduit during the bioremediation experiment
was similar in spring and autumn (mean value ± SD 7457.50 ± 53.03 cm3 ). The water
flow in the drainage conduit ranged between 15 and 28 L sec−1 before and after the tank
discharge, respectively.

2.3. Microbiological Analysis

Bacteriological analyses were performed on wastewater flowing in the drainage con-
duit where the sponge lives.
Wastewater was collected in triplicates at two sampling periods (spring and autumn)
for three collection times—immediately before (T0) and after (T1) the discharge of fish
tank wastewater and five minutes later (T2)—at two collection points: upstream (UP) and
downstream (DOWN) of the sponge biomass (Figure 1). Bacteriological analyses included
the quantitative analyses of cultivable vibrios, total culturable bacteria (37 ◦ C), fecal and
total coliforms, and fecal enterococci.
Enumeration of the culturable vibrios in seawater was achieved by filtering volumes
of 1, 5, and 10 mL of seawater on 0.22 µm pore size filters and aseptically placing the
filter disks onto thiosulphate-citrate-bile salt-sucrose agar (TCBS) plus 2% NaCl, a selective
J. Mar. Sci. Eng. 2022, 10, 874 4 of 13

medium for halophilic vibrio. Incubation was carried out at 25 ◦ C and 35 ◦ C for two days
and the colonies of presumptive vibrios were counted according to the colony-forming
unit (CFU) method. The fraction of vibrios that were potentially pathogenic to humans
J. Mar. Sci. Eng. 2022, 10, x FOR PEERwas estimated with the incubation temperature of 35 ◦ C. An incubation temperature
REVIEW 4 of 15
of
20–25 ◦ C was chosen because some Vibrio spp., such as Vibrio anguillarum, do not grow well
at higher temperatures [58]. Mean values for three replicate samples were determined.

Figure 1. Schematic drawing (not in scale) and real picture of the wastewater drainage conduit
Figure 1. Schematic drawing (not in scale) and real picture of the wastewater drainage conduit
where the sponge, Hymeniacidon perlevis, lives, as well as an indication of the wastewater collection
where the sponge, Hymeniacidon perlevis, lives, as well as an indication of the wastewater
points: upstream
collection points: (UP) and downstream
upstream (DOWN). In
(UP) and downstream the red box,
(DOWN). picts
In the redofbox,
H. perlevis with
picts of H. a polyvinyl
perlevis with
chloride (PVC) frames utilized for sponge volume measurement.
a polyvinyl chloride (PVC) frames utilized for sponge volume measurement.

The total culturable

2.3. Microbiological bacterial densities that grew at 37 ◦ C (indicating the fraction of
Analysis
bacteria potentially pathogenic to humans) were determined in the number of CFU by
Bacteriological
plating analysesand
100 µL of undiluted were performed
serial dilutionson wastewater
of seawater flowing in
in triplicate on the drainage
a plate count
conduit where the sponge lives. ◦
agar (PCA). The plates were incubated at 37 C for seven days [31]. All of the colonies
wereWastewater wasthe
counted using collected in triplicates
CFU method at two
through sampling
a 10x periods lens
magnification (spring andend
at the autumn)
of the
for three collection times—immediately before (T0) and after (T1) the discharge of fish
tank wastewater and five minutes later (T2)—at two collection points: upstream (UP) and
downstream (DOWN) of the sponge biomass (Figure 1). Bacteriological analyses included
the quantitative analyses of cultivable vibrios, total culturable bacteria (37 °C), fecal and
total coliforms, and fecal enterococci.
Enumeration of the culturable vibrios in seawater was achieved by filtering volumes
J. Mar. Sci. Eng. 2022, 10, 874 5 of 13

incubation period. For each microbiological parameter counted according to the CFU
method, the bacterial densities were expressed as CFU/mL.
Total and fecal coliforms as well as fecal enterococci were determined by the most
probable number (MPN) method using the standard five-tube method of 10-fold dilutions
for seawater samples [59]. For total and fecal coliform determination, lactose broth and
brilliant-green-lactose broth were used as cultural media in the presumptive and confirma-
tive tests, respectively. For fecal enterococci, the presumptive test was performed using
azide broth and the confirmative test was performed using ethyl violet azide broth. The
results are reported as MPN/100 mL.

2.4. Statistical Analysis

All analyses were performed using the Primer software and at a level of significance
of 95%. All experimental data were computed as dependent variables using PERMANOVA
in an approach similar to parametric ANOVA. Univariate PERMANOVA tests were run on
Bray–Curtis similarity matrices with 9999 permutations [60]. Season (S; 2 levels: autumn
and spring,), Station (St; 2 levels: UP and DOWN), and Time (T, 3 levels: T0, T1, and
T2) factors were used to detect the differences in the removal efficiency of Total Vibrio,
Total CFU, Fecal coliforms, Fecal enterococci, and Total coliforms. Each factor and its
interactions were individually analyzed using Univariate PERMANOVA tests with the
same experimental design. If necessary, square root transformed data in a Bray–Curtis
similarity matrix with 9999 permutations were used to perform the analyses [60]. If it was
impossible to obtain enough permutations for PERMANOVA analysis, the reference p was
obtained using a permutation simulation test (MONTECARLO test). The PAIR-WISE test
was applied to discover statistically significant differences in each pair of factor levels based
on the significance value of PERMANOVA/MONTECARLO tests.

3. Results
Microbiological analysis showed that Hymeniacidon perlevis in the drainage conduit is
able to reduce all the considered bacterial categories during both the spring and autumn
sampling period (Figures 2 and 3). With regards to culturable Vibrios, this category seems
to be the least favorite one by H. perlevis. In fact, in the upstream wastewater (T0_UP,
T1_UP, T2_UP; before the sponge) its value differs from that found in the corresponding
downstream wastewater (T0_DOWN, T1_DOWN, T2_DOWN; after the sponge), with
only a few hundred of CFU/mL—almost at each sampling time (T0, T1 and T2). The
vibrios concentration reduce at least an order of magnitude at T1 and T2, particularly
during the autumn sampling, when their concentration is the highest recorded in the whole
experiment (e.g., Autumn: T1_UP 8.90 ± 1.1 × 104 and T1_DOWN 4.50 ± 0.25 × 104 ;
T2_UP 1.75 ± 0.10 × 104 and T2_DOWN 4.55 ± 0.97 × 103 CFU mL−1 ; Spring: T2_UP
1.50 ± 0.11 × 103 and T2_DOWN 8.50 ± 0.55 × 102 CFU mL−1 ) (Figure 2). PERMANOVA
analyses showed significant differences in bacterial concentration before (UP) and after
(DOWN) the passage through the sponge community in the interaction between the three
factors considered (PERMANOVA, SE×ST×T, Pseudo-F = 12.591, p (perm) < 0.001). The
PAIR-WISE posterior test revealed significant differences in all three observation times in
the autumn period, while no significant domination was recorded in the spring for any T
factor level.
Total culturable bacteria at 37 ◦ C values in the downstream wastewater were always
lower than that found in the upstream wastewater, with an impressive reduction during
the autumn collection with a reduction of three and two orders of magnitude at T1 and T2,
respectively (Figure 2). The PERMANOVA analyses, carried out on the same experimental
design, also showed significant differences between UP and DOWN in the interaction be-
tween the three factors considered for this bacterial category (PERMANOVA, SE × St × T,
Pseudo-F = 69.264, p (perm) = 0.001). According to the PAIRWISE test, these differences
were significant for all levels of the analyzed factors, with the exception of T1 in the spring.
 considered (PERMANOVA, SE×ST×T, Pseudo-F = 12.591, p (perm) < 0.001). T
WISE posterior test revealed significant differences in all three observation tim
autumn period, while no significant domination was recorded in the spring
factor level.
J. Mar. Sci. Eng. 2022, 10, 874 6 of 13

Figure 2.Vibrios
Figure 2. Culturable Culturable Vibrios
(A) and (A) and bacteria
total culturable total culturable bacteria
at 37 ◦ C (B) at 37 °C
mean values (B) mean values
± standard
errors in wastewater upstream (UP) and downstream (DOWN) of
errors in wastewater upstream (UP) and downstream (DOWN) of Hymeniacidon perlevis at each Hymeniacidon perle
sampling time (T0, T1, T2) during both sampling periods
sampling time (T0, T1, T2) during both sampling periods (autumn and spring). (autumn and spring).
J.J.Mar.
Mar. Sci. Eng. 2022, 10, x FOR PEER REVIEW
Sci. Eng. 2022, 10, 874 7 of 15
7 of 13

Figure 3.
Figure 3. Fecal
Fecal contamination
contamination indicators
indicators (mean
(meanvalues
values±±standard
standarderrors)
errors)ininwastewater
wastewaterupstream
upstream
(UP) and downstream (DOWN) of the sponge Hymeniacidon perlevis at each sampling
(UP) and downstream (DOWN) of the sponge Hymeniacidon perlevis at each sampling time (T0,time (T0,T1,
T1, T2) during both sampling periods (autumn and spring).
T2) during both sampling periods (autumn and spring).
J. Mar. Sci. Eng. 2022, 10, 874 8 of 13

A similar trend was found regarding the fecal contamination indicators, where those
values were lower in the downstream wastewater compared with that in the upstream
wastewater for all three bacterial categories considered (total coliforms, fecal coliforms, fecal
enterococci, Figure 3). PERMANOVA highlighted a level of significance in the interaction
of the three factors (PERMANOVA, SE × ST × T, Pseudo-F = 13.258, p (perm) < 0.001).
The a posteriori test on the interaction of the three factors showed a significant reduction
in the concentration of coliforms at all times in the two seasons, with the exception of
T2 in the spring. In particular, in the autumn sampling, the total coliforms at T0 in the
upstream wastewater were 5.4 ± 0.98 × 102 MPN 100 mL−1 , while in the downstream
wastewater they reduced to 7.9 ± 1.1 × 10 MPN 100 mL−1 . Regarding fecal coliforms,
statistical analyses have highlighted a significance in the interaction between seasons and
stations (PERMANOVA, Pseudo-F = 5.2483, p (perm) = 0.0204) with significant differences
in the spring during the times T0 and T1. (Figure 3). Also, fecal enterococci in the upstream
wastewaters were always higher than the downstream ones with values that were lower
by, at least, an order of magnitude at each time analyzed (T0, T1, T2) (e.g., autumn: T1_UP
1.61 ± 0.23 × 103 MPN 100 mL−1 , and T1_DOWN 3.45 ± 0.19 × 102 MPN 100 mL−1 ;
spring: T1_UP 4.9 ± 0.28 × 10 MPN 100 mL−1 , and T1_DOWN 6.0 ± 0.17 MPN 100 mL−1
(Figure 3). These differences are clear in the PERMANOVA analysis, which highlights a
significance level in the interaction of the three factors (PERMANOVA, pseudo-F = 7.2591,
p (perm) < 0.001) and a significant decrease in the enterococci concentration at any time for
each season.

4. Discussion and Conclusions

Aquaculture plays a critical role in global food production, but with its explosive
expansion, considerable negative environmental impacts become obvious [61–63]. FAO
cautioned that such an increase will mainly be restricted by two factors, environmental
impacts and aquaculture diseases. One of the most prominent issues is that the water-borne
bacterial pathogens represent a significant biohazard for both aquaculture species and
human health [64]. To prevent pathogen-related aquaculture diseases and the potential
contamination of aquaculture products by pathogens, bioremediation is an attractive
option in aquaculture ecosystems. In particular, filter-feeder invertebrates, such as sponges,
polychaetes, and ascidians, due to their ability to remove pathogenic bacteria through the
filtration process, have recently been proposed as potential bioremediators [31,65–74].
The present study represents a significant advance on this topic since the demosponge
Hymeniacidon perlevis was employed in a land-based experimental fish farm as a novel
bioremediation technology to reduce the bacterial concentration and restore water quality.
Field and laboratory studies conducted in the Mediterranean area showed the ability
of some sponge species to remove, accumulate, and concentrate bacteria from the surround-
ing environment with higher efficiency for autochthonous bacteria, both under oligotrophic
and eutrophic conditions [29–33,35,36,75–77]. H. perlevis is able to remove bacteria even
in eutrophic environments thanks to its ability to tolerate high levels of microbial pol-
lution [32]. For this reason, this species was indicated as a potential bioremediator of
microbial pollution in impacted semi-enclosed basins [32].
The filtering activity of H. perlevis was also studied in laboratory experiments in which
its capability to remove and accumulate bacteria from sewage flowing into the Northern
Ionian Sea was investigated [31].
Present experiments showed that H. perlevis is able to filter and remove all the consid-
ered bacterial groups from the wastewater and this was reflected in the observed reduced
bacterial abundances in the out-flowing wastewater in comparison to those recorded in the
waste in-flowing system. Thus, H. perlevis can effectively control the growth of potential
pathogenic bacteria, including human pathogens culturable at 37 ◦ C, and vibrios in the
wastewater, with important epidemiological implications since its accumulation by these
filter-feeders may reduce their abundance in seawater and their risk as a biohazard to
human health.
J. Mar. Sci. Eng. 2022, 10, 874 9 of 13

The diffusion of filter-feeders, such as H. perlevis, in an aquaculture scenario could al-

low fish a better rearing environment, with lower risks related to the presence of pathogenic
bacteria such as vibrios, which are one of the main causes of infections and mass deaths
of farmed fish. Furthermore, this would also reduce human health risks and the financial
losses resulting from the loss of farmed fish products [78–80].
H. perlevis was particularly efficient in removing all the examined microbial pollution
indicators from the surrounding environment. The proven bioremediation capability of
the sponges when reared in an integrated system could be strategic since H. perlevis lives
in environments subjected to strong anthropogenic pollution. Thus, our results demon-
strated that in the presence of the sponge, the quality of seawater is improved, thereby
reaching low values of coliform bacteria and leading to new realms of bioremediation for
coastal communities.
The H. perlevis biomass living in the drainage conduit of the experimental fish farm
found in the present study seem to be able to reduce the bacterial concentration at both
wastewater flow values with an average percentage abatement for all bacterial categories
that ranges between 52% and 89% in the autumn and 35% and 93% in the spring. It has
been demonstrated that food availability affects the filtering physiology of sponges: With
the increase of food concentration, sponges respond with a decrease in pumping rate as
a protection against overloading the sponge feeding system, but the number of filtered
particles remains constant over time [81]. The results obtained in the present research
are in agreement with the abovementioned findings; in fact, in autumn, all the bacterial
categories considered, except for fecal coliforms, showed a higher concentration and a
lower abatement percentage with respect to those measured in spring.
The employment of H. perlevis in aquaculture waste treatment represents a more
attractive option to reduce bacterial loads in comparison to other technologies with high
negative impacts on the whole ecosystem. In particular, antibiotics-supplemented feeds
are commonly used in farms [82,83]. The massive use of antibiotics to control infections
in aquaculture has resulted in the development of resistant strains, which have rendered
antibiotic treatments ineffective.
In addition, the observation of biomass recovery of H. perlevis in the drainage conduits
of the fish farm indicated a surprising resilience of this species, opening a new scenario
on sponge rearing applications in land-based aquaculture facilities. We observed that
H. perlevis is ubiquitous in the drainage conduits of the experimental fish farm. Moreover,
although the drainage conduits were periodically cleaned by removing the sponge biomass,
it reaches impressive volume even after a few months (authors’ personal observation).
The use of marine sponges as bioremediators in appropriate environments is attractive
due to their effectiveness in reducing microbial pollution and the potential valorisation of
their biomass. Indeed, H. perlevis is known as a cytotoxic sponge and its associated marine
bacteria show antimicrobial and anticancer properties [84–86]. In addition, it has recently
been demonstrated that culturable bacteria associated with this sponge are promising
sources of antibacterial compounds of great pharmaceutical interest against pathogenic
multi-drug resistant strains (e.g., Staphylococcus aureus), which new antibiotics are in urgent
need of [87].
The H. perlevis rearing in land-based aquaculture facilities with bioremediation pur-
poses is also supported by the potential marketable value of the obtainable sponge biomass,
suggesting a novel application of zooremediation methodologies with potential commercial
gain. Thus, we propose an aquaculture farm model that is ecologically and economically
self-sustaining where, in one embodiment, it involves fish cultivation and sponge rearing.
Future studies should be aimed at investigating the effects of bioaccumulation of
microbial pollutants in H. perlevis and determine the tolerance limits for the selected species,
as well as the detection of biohazards in sponge tissues. The investigation of the capacity
for this sponge to utilise coliform bacteria may also lead to new realms of bioremediation
for coastal communities.
J. Mar. Sci. Eng. 2022, 10, 874 10 of 13

Author Contributions: Conceptualization, C.L. and L.S.; methodology, C.P. and L.S.; validation, C.L.
and L.S.; formal analysis, C.P. and L.S.; data curation, C.L. and R.T.; writing—original draft prepa-
ration, C.L., C.P. and L.S.; writing—review and editing, M.M., R.T., F.C., P.C. and S.A.; supervision,
C.L.; project administration, C.L.; funding acquisition, C.L. All authors have read and agreed to the
published version of the manuscript.
Funding: This research was partially funded by European Community, Life Environment funding
program: Remedia-Life project (project ID number: LIFE16 ENV/IT/000343) and partially funded by
Apulia Region PO FEAMP 2014–2020 Mis. 2.47: Approcci innovativi per una acquacoltura integrata e
sostenibile (project ID number: 003/INA/20).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Gaino, E.; Manconi, R.; Pronzato, R. Organizational plasticity as a successful conservative tactics in sponges. Anim. Biol. 1995, 4,
31–43.
2. Bosch, T.C.G.; Adamska, M.; Augustin, R.; Domazet-Loso, T.; Foret, S.; Fraune, S.; Funayama, N.; Grasis, J.; Hamada, M.; Hatta, M.
How do environmental factors influence life cycles and development? An experimental framework for early-diverging metazoans.
Bioessays 2014, 36, 1185–1194. [CrossRef] [PubMed]
3. Ribes, M.; Coma, R.; Gili, J.M. Natural diet and grazing rate of the temperate sponge Dysidea avara (Demospongiae, Dendrocer-
atida) throughout an annual cycle. Mar. Ecol. Prog. Ser. 1999, 176, 179–190. [CrossRef]
4. Pile, A.J.; Patterson, M.R.; Witman, J.D. In situ grazing on plankton <10 µm by the boreal sponge Mycale lingua. Mar. Ecol. Prog.
Ser. 1996, 141, 95–102. [CrossRef]
5. de Goeij, J.M.; van den Berg, H.; van Oostveen, M.M.; Epping, E.H.G.; Van Duyl, F.C. Major bulk dissolved organic carbon (DOC)
removal by encrusting coral reef cavity sponges. Mar. Ecol. Prog. Ser. 2008, 357, 139–151. [CrossRef]
6. Simpson, T.L. The Cell Biology of Sponges, 1st ed.; Springer: New York, NY, USA, 1984; ISBN 978-1-4612-9740-6.
7. Corriero, G.; Longo, C.; Mercurio, M.; Nonnis Marzano, C.; Lembo, G.; Spedicato, M.T. Rearing performance of Spongia officinalis
on suspended ropes off the Southern Italian Coast (Central Mediterranean Sea). Aquaculture 2004, 238, 195–205. [CrossRef]
8. Duckworth, A.R.; Wolff, C. Bath sponge aquaculture in Torres Strait, Australia: Effect of explant size, farming method and the
environment on culture success. Aquaculture 2007, 271, 188–195. [CrossRef]
9. Duckworth, R.A. The role of behavior in evolution: A search for mechanism. Evol. Ecol. 2009, 23, 513–531. [CrossRef]
10. Bierwirth, J.; Mantas, T.P.; Villechanoux, J.; Cerrano, C. Restoration of marine sponges-What can we learn from over a century of
experimental cultivation? Water 2022, 14, 1055. [CrossRef]
11. Cao, H.; Xue, L.; Cao, X.; Zhang, W. Laboratory adaptation of life cycle of Hymeniacidon perlevis explants cultured in aquarium. J.
Biotechnol. 2008, 136, S551. [CrossRef]
12. Xue, L.; Zhang, W. Growth and survival of early juveniles of the marine sponge Hymeniacidon perlevis (Demospongiae) under
controlled conditions. Mar. Biotechnol. 2009, 11, 640–649. [CrossRef] [PubMed]
13. Xue, L.; Zhang, X.; Zhang, W. Larval release and settlement of the marine sponge Hymeniacidon perlevis (Porifera, Demospongiae)
under controlled laboratory conditions. Aquaculture 2009, 290, 132–139. [CrossRef]
14. Maslin, M.; Gaertner-Mazouni, N.; Debitus, C.; Joy, N.; Ho, R. Marine sponge aquaculture towards drug development: An
ongoing history of technical, ecological, chemical considerations and challenges. Aquac. Rep. 2021, 21, 100813. [CrossRef]
15. Binnewerg, B.; Schubert, M.; Voronkina, A.; Muzychka, L.; Wysokowski, M.; Petrenko, I.; Djurović, M.; Kovalchuk, V.; Tsurkan, M.;
Martinovic, R. Marine biomaterials: Biomimetic and pharmacological potential of cultivated Aplysina aerophoba marine demo-
sponge. Mater. Sci. Eng. C 2020, 109, 110566. [CrossRef]
16. Pita, L.; Rix, L.; Slaby, B.M.; Franke, A.; Hentschel, U. The sponge holobiont in a changing ocean: From microbes to ecosystems.
Microbiome 2018, 6, 46. [CrossRef]
17. Slaby, B.M.; Franke, A.; Rix, L.; Pita, L.; Bayer, K.; Jahn, M.T.; Hentschel, U. Marine sponge holobionts in health and disease. In
Symbiotic Microbiomes of Coral Reefs Sponges and Corals; Li, Z., Ed.; Springer: Dordrecht, The Netherlands, 2019; pp. 81–104, ISBN
978-94-024-1610-7.
18. Schellenberg, J.; Reichert, J.; Hardt, M.; Klingelhöfer, I.; Morlock, G.; Schubert, P.; Bižić, M.; Grossart, H.-P.; Kämpfer, P.; Wilke,
T. The bacterial microbiome of the long-term aquarium cultured high-microbial abundance sponge Haliclona cnidata–sustained
bioactivity despite community shifts under detrimental conditions. Front. Mar. Sci. 2020, 7, 266. [CrossRef]
19. Bastaraud, A.; Cecchi, P.; Handschumacher, P.; Altmann, M.; Jambou, R. Urbanization and waterborne pathogen emergence in
low-income countries: Where and how to conduct surveys? Int. J. Environ. Res. Public Health 2020, 17, 480. [CrossRef]
J. Mar. Sci. Eng. 2022, 10, 874 11 of 13

20. Landrigan, P.J.; Stegeman, J.J.; Fleming, L.E.; Allemand, D.; Anderson, D.M.; Backer, L.C.; Brucker-Davis, F.; Chevalier, N.;
Corra, L.; Czerucka, D.; et al. Human Health and Ocean Pollution. Ann. Glob. Health 2020, 86, 151. [CrossRef]
21. Reilly, A.; Käferstein, F. Food safety hazards and the application of the principles of the hazard analysis and critical control point
(HACCP) system for their control in aquaculture production. Aquac. Res. 1997, 28, 735–752. [CrossRef]
22. Saxena, G.; Kishor, R.; Bharagava, R.N. Bioremediation of Industrial Waste for Environmental Safety, 1st ed.; Gaurav Saxena, R.N.B.,
Ed.; Springer: Singapore, 2020; ISBN 978-981-13-1890-0.
23. Marimuthu, P.D.; Sundaram, R.; Jeyaseelan, A.; Kaliannan, T. (Eds.) Bioremediation and Green Technologies, 1st ed.; Environmental
Science and Engineering; Springer International Publishing: Cham, Switzerland, 2021; ISBN 978-3-030-64121-4.
24. Suyal, D.C.; Soni, R.; Eds. Bioremediation of Environmental Pollutants: Emerging Trends and Strategies, 1st ed.; Springer International
Publishing: Cham, Switzerland, 2022; ISBN 978-3-030-86168-1.
25. Gifford, S.; Dunstan, R.H.; O’Connor, W.; Koller, C.E.; MacFarlane, G.R. Aquatic zooremediation: Deploying animals to remediate
contaminated aquatic environments. Trends Biotechnol. 2007, 25, 60–65. [CrossRef]
26. Fu, W.; Sun, L.; Zhang, X.; Zhang, W. Potential of the marine sponge Hymeniacidon perleve as a bioremediator of pathogenic
bacteria in integrated aquaculture ecosystems. Biotechnol. Bioeng. 2006, 93, 1112–1122. [CrossRef] [PubMed]
27. Zhang, X.; Zhang, W.; Xue, L.; Zhang, B.; Jin, M.; Fu, W. Bioremediation of bacteria pollution using the marine sponge Hymeniacidon
perlevis in the intensive mariculture water system of turbot Scophthalmus maximus. Biotechnol. Bioeng. 2010, 105, 59–68. [CrossRef]
28. Huysecom, J.; Richelle-Maurer, E.; Van De Vyver, G.; Vray, B. Effect of bacterial concentration on retention and growth rate of the
freshwater sponge Ephydatia fluviatilis. Physiol. Zool. 1988, 61, 535–542. [CrossRef]
29. Stabili, L.; Licciano, M.; Giangrande, A.; Longo, C.; Mercurio, M.; Marzano, C.N.; Corriero, G. Filtering activity of Spongia
officinalis var. adriatica (Schmidt) (Porifera, Demospongiae) on bacterioplankton: Implications for bioremediation of polluted
seawater. Water Res. 2006, 40, 3083–3090. [CrossRef] [PubMed]
30. Stabili, L.; Licciano, M.; Longo, C.; Corriero, G.; Mercurio, M. Evaluation of microbiological accumulation capability of the
commercial sponge Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae). Water Res. 2008, 42, 2499–2506. [CrossRef]
31. Longo, C.; Corriero, G.; Licciano, M.; Stabili, L. Bacterial accumulation by the Demospongiae Hymeniacidon perlevis: A tool for the
bioremediation of polluted seawater. Mar. Pollut. Bull. 2010, 60, 1182–1187. [CrossRef]
32. Longo, C.; Cardone, F.; Corriero, G.; Licciano, M.; Pierri, C.; Stabili, L. The co-occurrence of the demosponge Hymeniacidon perlevis
and the edible mussel Mytilus galloprovincialis as a new tool for bacterial load mitigation in aquaculture. Environ. Sci. Pollut. Res.
2016, 23, 3736–3746. [CrossRef]
33. Trani, R.; Corriero, G.; De Pinto, M.C.; Pierri, C.; Scrascia, M.; Longo, C.; Mercurio, M.; Pazzani, C. Filtering activity and nutrient
release by the keratose sponge Sarcotragus spinosulus Schmidt, 1862 (Porifera, Demospongiae) at the laboratory scale. J. Mar. Sci.
Eng. 2021, 9, 178. [CrossRef]
34. Maldonado, M.; Zhang, X.; Cao, X.; Xue, L.; Cao, H.; Zhang, W. Selective feeding by sponges on pathogenic microbes: A
reassessment of potential for abatement of microbial pollution. Mar. Ecol. Prog. Ser. 2010, 403, 75–89. [CrossRef]
35. Ledda, F.D.; Manconi, R.; Pronzato, R. Retention rates on bacteria and organic matter by Ircinia variabilis (Demospongiae,
Dictyoceratida) in experimental sponge farming for bioremediation. Biol. Mar. Mediterr. 2008, 15, 164–165.
36. Ledda, F.D.; Pronzato, R.; Manconi, R. Mariculture for bacterial and organic waste removal: A field study of sponge filtering
activity in experimental farming. Aquac. Res. 2014, 45, 1389–1401. [CrossRef]
37. Hansen, I.V.; Weeks, J.M.; Depledge, M.H. Accumulation of copper, zinc, cadmium and chromium by the marine sponge
Halichondria panicea Pallas and the implications for biomonitoring. Mar. Pollut. Bull. 1995, 31, 133–138. [CrossRef]
38. Mahaut, M.L.; Basuyaux, O.; Baudinière, E.; Chataignier, C.; Pain, J.; Caplat, C. The porifera Hymeniacidon perlevis (Montagu,
1818) as a bioindicator for water quality monitoring. Environ. Sci. Pollut. Res. 2013, 20, 2984–2992. [CrossRef] [PubMed]
39. Santos-Gandelman, J.F.; Cruz, K.; Crane, S.; Muricy, G.; Giambiagi-deMarval, M.; Barkay, T.; Laport, M.S. Potential application in
mercury bioremediation of a marine sponge-isolated Bacillus cereus strain Pj1. Curr. Microbiol. 2014, 69, 374–380. [CrossRef]
40. Gentric, C.; Rehel, K.; Dufour, A.; Sauleau, P. Bioaccumulation of metallic trace elements and organic pollutants in marine sponges
from the South Brittany Coast, France. J. Environ. Sci. Health-Part A Toxic/Hazardous Subst. Environ. Eng. 2016, 51, 213–219.
[CrossRef]
41. Ferrante, M.; Vassallo, M.; Mazzola, A.; Brundo, M.V.; Pecoraro, R.; Grasso, A.; Copat, C. In vivo exposure of the marine sponge
Chondrilla nucula Schmidt, 1862 to cadmium (Cd), copper (Cu) and lead (Pb) and its potential use for bioremediation purposes.
Chemosphere 2018, 193, 1049–1057. [CrossRef]
42. Orani, A.M.; Barats, A.; Zitte, W.; Morrow, C.; Thomas, O.P. Comparative study on the bioaccumulation and biotransformation of
arsenic by some northeastern Atlantic and northwestern Mediterranean sponges. Chemosphere 2018, 201, 826–839. [CrossRef]
43. Gravina, M.F.; Longo, C.; Puthod, P.; Rosati, M.; Colozza, N.; Scarselli, M. Heavy metal accumulation capacity of Axinella
damicornis (Esper, 1794)(Porifera, Demospongiae): A tool for bioremediation of polluted seawaters. Mediterr. Mar. Sci. 2022, 23,
125–133. [CrossRef]
44. Yang, Q.; Franco, C.M.M.; Zhang, W. Uncovering the hidden marine sponge microbiome by applying a multi-primer approach.
Sci. Rep. 2019, 9, 6214. [CrossRef]
45. Nicholson, J.K.; Holmes, E.; Kinross, J.; Burcelin, R.; Gibson, G.; Jia, W.; Pettersson, S. Host-Gut Microbiota Metabolic Interactions.
Science 2012, 336, 1262–1267. [CrossRef]
J. Mar. Sci. Eng. 2022, 10, 874 12 of 13

46. Zhang, X.; Zhang, W.; Xue, L.; Zhang, B. Feasibility of sponge Hymeniacidon perlevis in controlling pathogenic bacteria in turbot
Scophthalmus maximus-culture water system. J. Biotechnol. 2008, 136, S685. [CrossRef]
47. Aresta, A.; Marzano, C.N.; Lopane, C.; Corriero, G.; Longo, C.; Zambonin, C.; Stabili, L. Analytical investigations on the lindane
bioremediation capability of the demosponge Hymeniacidon perlevis. Mar. Pollut. Bull. 2015, 90, 143–149. [CrossRef] [PubMed]
48. Stabili, L.; Pizzolante, G.; Morgante, A.; Nonnis Marzano, C.; Longo, C.; Aresta, M.A.; Zambonin, C.; Corriero, G.; Alifano, P.
Lindane bioremediation capability of bacteria associated with the demosponge Hymeniacidon perlevis. Mar. Drugs 2017, 15, 108.
49. Gaino, E.; Frine, C.; Giuseppe, C. Reproduction of the intertidal sponge Hymeniacidon perlevis (Montagu) along a bathymetric
gradient. Open Mar. Biol. J. 2010, 4, 47–56. [CrossRef]
50. Longo, C.; Altınsaçlı, S.; Yardımcı, C.H.; Altınsaçlı, S. Occurrence of Hymeniacidon perlevis (Montagu, 1814) (Porifera, Demospon-
giae) in Kamil Abduş Lagoon, İstanbul, Turkey. J. Entomol. Zool. Stud. 2017, 5, 1620–1625.
51. Erpenbeck, D.; Van Soest, R.W.M. Family Halichondriidae Gray, 1867. In Systema Porifera; Hooper, J.N.A., Van Soest, R.W.M.,
Willenz, P., Eds.; Springer: Boston, MA, USA, 2002; pp. 787–815, ISBN 978-0-306-47260-2.
52. de Voogd, N.; Alvarez, B.; Boury-Esnault, N.; Carballo, J.; Cárdenas, P.; Díaz, M.-C.; Dohrmann, M.; Downey, R.; Hajdu, E.;
Hooper, J.; et al. Hymeniacidon Perlevis (Montagu, 1814). Available online: http://www.marinespecies.org/porifera/porifera.
php?p=taxdetails&id=132663 (accessed on 25 April 2022).
53. Turner, T. The marine sponge Hymeniacidon perlevis is a globally-distributed exotic species. Aquat. Invasions 2020, 15, 542–561.
[CrossRef]
54. Cabioch, L. Contribution à la connaissance de la faune des Spongiaires de la Manche occidentale. Démosponges de la région de
Roscoff. Cah. Biol. Mar. 1968, 9, 211–246.
55. Longo, C.; Cardone, F.; Mercurio, M.; Marzano, C.N.; Pierri, C.; Corriero, G. Spatial and temporal distributions of the sponge
fauna insouthern Italian lagoon systems. Mediterr. Mar. Sci. 2016, 17, 174–189. [CrossRef]
56. Juniper, A.J.; Steele, R.D. Intertidal sponges of the Portsmouth area. J. Nat. Hist. 1969, 3, 153–163. [CrossRef]
57. Stone, A.R. Growth and reproduction of Hymeniacidon perleve (Montagu) (Porifera) in langstone harbour hampshire. J. Zool. 1970,
161, 443–459. [CrossRef]
58. Høi, L.; Larsen, J.L.; Dalsgaard, I.; Dalsgaard, A. Occurrence of Vibrio vulnificus biotypes in Danish marine environments. Appl.
Environ. Microbiol. 1998, 64, 7–13. [CrossRef] [PubMed]
59. Clesceri, S.L.; Greenberg, E.A.; Eaton, A.D. (Eds.) Standard Methods for Examination of Water and Wastewater, 20th ed.; American
Public Health Association, American Water Works Association, Water Environment Federation: Washington, DC, USA, 1998;
ISBN 9780875532875.
60. Anderson, M.J. A new method for non-parametric multivariate analysis of variance. Austral Ecol. 2001, 26, 32–46. [CrossRef]
61. FAO. The State of World Fisheries and Aquaculture 2020: Sustainability in Action; FAO: Rome, Italy, 2020; ISBN 978-92-5-132692-3.
62. Halpern, B.S.; Walbridge, S.; Selkoe, K.A.; Kappel, C.V.; Micheli, F.; D’Agrosa, C.; Bruno, J.F.; Casey, K.S.; Ebert, C.; Fox, H.E.; et al.
A global map of human impact on marine ecosystems. Science 2008, 319, 948–952. [CrossRef] [PubMed]
63. Halpern, B.S.; Frazier, M.; Afflerbach, J.; Lowndes, J.S.; Micheli, F.; O’Hara, C.; Scarborough, C.; Selkoe, K.A. Recent pace of
change in human impact on the world’s ocean. Sci. Rep. 2019, 9, 11609. [CrossRef] [PubMed]
64. Cavallo, R.A.; Stabili, L. Culturable vibrios biodiversity in the Northern Ionian Sea (Italian coasts). Sci. Mar. 2004, 68, 23–29.
[CrossRef]
65. Burge, C.A.; Closek, C.J.; Friedman, C.S.; Groner, M.L.; Jenkins, C.M.; Shore-Maggio, A.; Welsh, J.E. The use of filter-feeders to
manage disease in a changing world. Integr. Comp. Biol. 2016, 56, 573–587. [CrossRef]
66. Stabili, L.; Licciano, M.; Longo, C.; Lezzi, M.; Giangrande, A. The Mediterranean non-indigenous ascidian Polyandrocarpa
zorritensis: Microbiological accumulation capability and environmental implications. Mar. Pollut. Bull. 2015, 101, 146–152.
[CrossRef]
67. Milanese, M.; Chelossi, E.; Manconi, R.; Sarà, A.; Sidri, M.; Pronzato, R. The marine sponge Chondrilla nucula Schmidt, 1862 as an
elective candidate for bioremediation in integrated aquaculture. Biomol. Eng. 2003, 20, 363–368. [CrossRef]
68. MMO. Evidence Supporting the Use of Environmental Remediation to Improve Water Quality in the South Marine Plan Areas; Marine
Management Organisation: Newcastle, UK, 2016.
69. Giangrande, A.; Cavallo, A.; Licciano, M.; Mola, E.; Pierri, C.; Trianni, L. Utilization of the filter feeder polychaete Sabella. Aquac.
Int. 2005, 13, 129–136. [CrossRef]
70. Giangrande, A.; Gravina, M.F.; Rossi, S.; Longo, C.; Pierri, C. Aquaculture and restoration: Perspectives from Mediterranean sea
experiences. Water 2021, 13, 991. [CrossRef]
71. Stabili, L.; Schirosi, R.; Licciano, M.; Mola, E.; Giangrande, A. Bioremediation of bacteria in aquaculture waste using the polychaete
Sabella spallanzanii. New Biotechnol. 2010, 27, 774–781. [CrossRef] [PubMed]
72. Licciano, M.; Stabili, L.; Giangrande, A.; Cavallo, R.A. Bacterial accumulation by Branchiomma luctuosum (Annelida: Polychaeta):
A tool for biomonitoring marine systems and restoring polluted waters. Mar. Environ. Res. 2007, 63, 291–302. [CrossRef] [PubMed]
73. Marques, L.; Calado, R.; Lillebø, A.I. Potential of ascidians as extractive species and their added value in marine integrated
multitrophic aquaculture systems–From pests to valuable blue bioresources. Front. Mar. Sci. 2022, 9, 129. [CrossRef]
74. Stabili, L.; Licciano, M.; Gravina, M.F.; Giangrande, A. Filtering activity on a pure culture of Vibrio alginolyticus by the solitary
ascidian Styela plicata and the colonial ascidian Polyandrocarpa zorritensis: A potential service to improve microbiological seawater
quality economically. Sci. Total Environ. 2016, 573, 11–18. [CrossRef]
J. Mar. Sci. Eng. 2022, 10, 874 13 of 13

75. Gökalp, M.; Mes, D.; Nederlof, M.; Zhao, H.; Merijn de Goeij, J.; Osinga, R. The potential roles of sponges in integrated mariculture.
Rev. Aquac. 2021, 13, 1159–1171. [CrossRef]
76. Trani, R.; de Pinto, M.C.; Scrascia, M.; Pierri, C.; Mercurio, M.; Marzano, C.N.; Deflorio, M.; Longo, C. Multi-trophic species
interactions in a marine bioremediation model assessment. In Proceedings of the 2021 International Workshop on Metrology for
the Sea; Learning to Measure Sea Health Parameters (MetroSea), Reggio Calabria, Italy, 4–6 October 2021; IEEE: Reggio Calabria,
Italy, 2021; pp. 312–315.
77. Longo, C.; Corriero, G.; Mercurio, M.; Licciano, M.; Stabili, L. Studio comparativo dell’accumulo microbiologico in due
demosponge mediterranee. Biol. Mar. Mediterr. 2008, 15, 424–425.
78. de Souza Valente, C.; Wan, A.H.L. Vibrio and major commercially important vibriosis diseases in decapod crustaceans. J. Invertebr.
Pathol. 2021, 181, 107527. [CrossRef]
79. Harrison, J.; Nelson, K.; Morcrette, H.; Morcrette, C.; Preston, J.; Helmer, L.; Titball, R.W.; Butler, C.S.; Wagley, S. The increased
prevalence of Vibrio species and the first reporting of Vibrio jasicida and Vibrio rotiferianus at UK shellfish sites. Water Res. 2022,
211, 117942. [CrossRef]
80. Bondad-Reantaso, M.G.; Subasinghe, R.P.; Arthur, J.R.; Ogawa, K.; Chinabut, S.; Adlard, R.; Tan, Z.; Shariff, M. Disease and health
management in Asian aquaculture. Vet. Parasitol. 2005, 132, 249–272. [CrossRef]
81. Morganti, T.M.; Ribes, M.; Yahel, G.; Coma, R. Size is the major determinant of pumping rates in marine sponges. Front. Physiol.
2019, 10, 1474. [CrossRef]
82. Done, H.Y.; Venkatesan, A.K.; Halden, R.U. Does the recent growth of aquaculture create antibiotic resistance threats different
from those associated with land animal production in agriculture? AAPS J. 2015, 17, 513–524. [CrossRef] [PubMed]
83. Chen, J.; Sun, R.; Pan, C.; Sun, Y.; Mai, B.; Li, Q.X. Antibiotics and food safety in aquaculture. J. Agric. Food Chem. 2020, 68,
11908–11919. [CrossRef] [PubMed]
84. Xue, S.; Zhang, H.T.; Wu, P.C.; Zhang, W.; Yuan, Q. Study on bioactivity of extracts from marine sponges in Chinese Sea. J. Exp.
Mar. Biol. Ecol. 2004, 298, 71–78. [CrossRef]
85. Zheng, L.; Yan, X.; Xu, J.; Chen, H.; Lin, W. Hymeniacidon perleve associated bioactive bacterium Pseudomonas sp. NJ6-3-1. Appl.
Biochem. Microbiol. 2005, 41, 29–33. [CrossRef]
86. Zheng, L.; Yan, X.; Han, X.; Chen, H.; Lin, W.; Lee, F.S.C.; Wang, X. Identification of norharman as the cytotoxic compound
produced by the sponge Hymeniacidon perleve-associated marine bacterium Pseudoalteromonas piscicida and its apoptotic effect on
cancer cells. Biotechnol. Appl. Biochem. 2006, 44, 135–142. [PubMed]
87. Rodriguez Jimenez, A.; Dechamps, E.; Giaux, A.; Goetghebuer, L.; Bauwens, M.; Willenz, P.; Flahaut, S.; Laport, M.S.; George, I.F.
The sponges Hymeniacidon perlevis and Halichondria panicea are reservoirs of antibiotic-producing bacteria against multi-drug
resistant Staphylococcus aureus. J. Appl. Microbiol. 2021, 131, 706–718. [CrossRef] [PubMed]