Genetic diversity in mitochondrial DNA reveals the effect of a Fisheries Protection Zone on exploited marine species in the Menorca Channel (Western Mediterranean)

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Ferragut, Maria Teresa Farriols, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7271616/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Overexploitation can drive evolutionary changes and erode genetic diversity, reducing species’ adaptive capacity to environmental and anthropogenic pressures. Spatial marine conservation measures, such as Marine Protected Areas and Fisheries Protection Zones (FPZs), aim to mitigate these impacts by preserving biodiversity and promoting sustainable fisheries. Recently, nucleotide diversity of the mitochondrial Cytochrome C Oxidase subunit I (COI) marker has emerged as a promising proxy for assessing species conservation status. To evaluate the effectiveness of an FPZ established in 2016 in the Menorca Channel, COI genetic diversity was assessed in four exploited marine species across three areas: the FPZ and two nearby non-protected zones. All species exhibited consistently higher genetic diversity within the FPZ, despite evidence of high gene flow among areas. Coalescent simulations were used to model expected genetic diversity under neutral scenarios of bottlenecks and expansions, with magnitudes estimated from differences in nucleotide diversities observed between fished and non-fished zones. Simulations supported a scenario of population expansion in the FPZ, contrasting with signs of genetic erosion in fished areas. These patterns align with Vessel Monitoring System (VMS) data, which show a post-protection-establishment shift in fishing effort toward non-protected zones, potentially contributing to population declines outside the FPZ. This study provides genetic evidence of the positive effects of fishing restrictions on fishery resources in the Menorca Channel, supporting the FPZ’s role in preserving genetic diversity and promoting population recovery. Furthermore, it highlights COI nucleotide diversity as a simple, cost-effective tool for monitoring marine species’ conservation status and guiding resource management strategies. Nucleotide diversity COI Coalescent simulations Fisheries resources Protected areas Mediterranean Sea Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction In the last few decades, the implementation of marine protected areas (MPAs) and fisheries protection zones (FPZs) has increased worldwide, implemented as tools to simultaneously achieve both biodiversity conservation and fisheries management objectives (Hilborn 2004 ; Gaines et al. 2010 ). Protected areas play an important role in fisheries management due to two main ecological mechanisms: the dispersal of adults and juveniles, defined as "spill-over" (Rowley 1994 ), and the larval supply. These areas contribute to the restoration of overfished fish stocks and provide net benefits to nearby areas (Di Lorenzo et al. 2016 , 2020 ; Andrello et al. 2017 ). In the Mediterranean Sea, a clear increase in biomass and abundance of exploited marine species within protected areas has been observed, with consequent benefits for fishing activities in areas adjacent to the reserves (Guidetti and Sala 2007 ; Harmelin-Vivien et al. 2008 ; Follesa et al. 2011 ; Giakoumi et al. 2017 ). These benefits have also been observed in individual fish conditions, assessed through physiological indicators (Lloret and Planes 2003 ; Lloret et al. 2005 ; Sillero-Ríos et al. 2018 ). These findings suggest that some fish species show a preference for habitats with better conditions for development, survival, feeding, and reproduction. Furthermore, it is known that the exploitation of fishery resources can affect the evolutionary dynamics of fish populations in different ways: i) loss of genetic diversity or adaptive potential (Pinsky and Palumbi 2014 ; Petit-Marty et al. 2022 ); ii) variations in the power of detection of population structure (Gandra et al. 2020 ); and iii) evolutionary changes due to fishing pressures (Allendorf et al. 2008 ). Intense fishing pressure leads to population declines, which will significantly reduce the levels of genetic diversity if declines are strong enough to produce changes in the frequency of alleles within the affected populations (Hauser et al. 2002 ; Hutchinson et al. 2003 ; Ruggeri et al. 2016 ). Since genetic variation is the basis of natural selection, its degradation can reduce the ability of a species to adapt to changing environments produced directly or indirectly by anthropogenic pressures, increasing the risk of local extinction (Spielman et al. 2004 ; Allendorf et al. 2008 ; Vásquez et al. 2023 ). In this sense, overfishing drives population decline, which in turn causes the loss of genetic diversity in exploited species and, therefore, potentially affects their conservation and adaptive capacity to environmental changes (Petit-Marty et al. 2022 ; Sadler et al. 2023 ; Mendoza-Portillo et al. 2023 ). In this context, mitochondrial genes can provide information not only on demographic changes and declines in population sizes but can also serve as diagnostic tools for assessing the conservation status of exploited populations (Johnson et al. 2018 ; Petit-Marty et al. 2020 ; Righi et al. 2020 ; Ilham Syahadah Mohd Yusoff et al. 2021; Canteri et al. 2021 ; Ferragut-Perello et al. 2023 ; Zlateva et al. 2023 ). Recently, some studies have demonstrated that estimates of genetic diversity—particularly the nucleotide diversity index—derived from the mitochondrial marker Cytochrome C Oxidase subunit I (COI) can be effective, simple and cost-efficient indicators of the conservation status of commercially exploited (Yorisue et al. 2020 ; Petit-Marty et al. 2022 ) and vulnerable (Ham-Dueñas et al. 2020 ; Petit-Marty et al. 2020 ; Ferragut-Perello et al. 2023 ) marine fish species. The genetic diversity observed in mitochondrial markers, combined with coalescent simulations, also provides insights into the demographic history of marine species over time. Coalescent simulations are a well-established method for generating population samples under different evolutionary scenarios (Ewing and Hermisson 2010 ) and have been used to estimate expected decreases in genetic diversity—under a neutral model of evolution—resulting from population declines driven by sustained harvesting pressure (Pinsky and Palumbi 2014 ; Arenas 2019 ; Petit-Marty et al. 2022 ; Reid and Pinsky 2022 ). The present study was carried out in the Menorca Channel, which is located between the islands of Mallorca and Menorca in the Western Mediterranean and represents nearly 20% of the coastal continental shelf of the Balearic Archipelago (40 to 100 m depth). Due to the hydrodynamic conditions, the area hosts a wide distribution of habitats and species of conservation interest, such as coralligenous outcrops, maërl, and biogenic detrital beds (Barberá et al. 2012 ; Moranta et al. 2014 ; Farriols et al. 2025 ). Red algae beds, composed of both Peyssonnelia and rhodoliths, have a high biodiversity and secondary macrobenthic production, which positively influence the abundance, physiological status, and key vital aspects of demersal resources on the Balearic continental shelf (Ordines et al. 2009 , 2015 ). Maërl and coralligenous beds are classified as sensitive habitats (HSs) and essential fish habitats (EFHs) (Ardizzone 2006 ). Following the implementation of Council Regulation (EC) No. 1967/2006, which establishes management measures for the sustainable exploitation of fishery resources in the Mediterranean Sea, rhodolith beds and coralligenous bottoms were recognized as protected habitats. Owing to the presence of these essential habitats and their spatial overlap with trawling activities and associated impacts, the Menorca Channel was declared a Site of Community Importance (SCI) in 2014. In 2016, trawling was subsequently banned in certain areas within the SCI, designated as Fisheries Protection Zone (FPZ). This study aims to assess the effect of a Fisheries Protection Zone (FPZ) on the conservation status of exploited marine species by analysing genetic diversity levels using the mitochondrial COI marker, comparing the FPZ located in the Menorca Channel with the surrounding areas subjected to fishing activity. To do this, three species of teleosts were used as models: Mullus surmuletus (red striped mullet), Serranus cabrilla (comber) and Scorpaena notata (small red scorpionfish), and one cephalopod species: Octopus vulgaris (common octopus). All these species are widely distributed throughout the East Atlantic and the Mediterranean Sea, inhabiting the narrow continental shelf areas characterized by rocky or sandy substrates and seagrass meadows (Ordines et al. 2009 ; Alós et al. 2011 ; Fadhlaoui-Zid et al. 2012 ; Félix-Hackradt et al. 2013 ; Bos et al. 2020 ; Pérez et al. 2023 ). Furthermore, in the Balearic Islands all species are commercially exploited, whether they are target species ( M. surmuletus , S. cabrilla, and O. vulgaris ) or bycatches ( S. notata ) (Quetglas et al. 1998 ; 2016 ). Materials and methods Sampling The samples of the studied species were collected during the MEDITS (International bottom trawl survey in the Mediterranean) and CANAL scientific surveys, which were partially or totally conducted in the Menorca Channel between 2021 and 2023. In these surveys, the experimental bottom trawl GOC-73 was used to sample demersal communities and resources from seafloor areas subject to fishing activity (for specific information see Spedicato et al. 2019). Specifically, the samples were obtained from FPZs located in the Menorca Channel and from surrounding fished area within the SCI and adjacent areas (ADJ), located south of the Menorca Channel. Sampling sites for the studied species are indicated in Fig. 1. For all the studied species, tissue samples were collected from at least 30 specimens per zone (FPZ, SCI, and ADJ), preserved in 96% ethanol and stored at -20ºC. The total number of tissue samples is indicated in Table 1. Fishing footprint Data from the Vessel Monitoring by satellite System (VMS) collected between 2009 and 2023, were analysed to assess the distribution of bottom trawl fishing efforts along the Menorca Channel before and after the establishment of the FPZ. Prior to analysis, VMS data were filtered to exclude signals produced during sailing and manoeuvring. To do so, only VMS signals recorded between 05:00 am and 05:00 pm (the period when trawlers are allowed to fish in the Balearic Islands) and with an instantaneous speed from 2 to 3.6 knots (the velocity range during fishing operations in the Balearic Islands) were considered. Finally, the mean annual number of VMS signals per cell in a 0.01º resolution grid was mapped with ArcGIS Desktop 10.8 (ESRI 2020) for the periods 2009-2015 and 2017-2023, corresponding to the sampling periods before and after the FPZ implementation, respectively. Laboratory procedures Total genomic DNA (gDNA) was extracted using the DNeasy Blood and Tissue kit (Qiagen, West Sussex, UK). Polymerase chain reaction (PCR) was used to amplify the genetic marker Cytochrome C Oxidase subunit I (COI) using different primer sets: FF2d/FR1d (600 bp for M. surmuletus , 642 bp for S. notata ;Ivanova et al. 2007), FishF1/FishR1 (612 bp for S. cabrilla ; Ward et al. 2005), and LCO1490/HCO2198 (657 bp for O. vulgaris ; Folmer et al. 1994). PCR reactions were carried out in a 25 μL volume containing: 2.5 μL of 10X Buffer (BIOLINE, London, UK), 1.75 μL of 50 mM MgCl 2 , 1 μL of 100 mM dNTPs, 0.5 μL of each primer (200 mM), 0.05 μL of DNA polymerase (BIOLINE) (5 U/μL), and 1 μL of gDNA template. PCR program conditions for each species and primer set are detailed in Table S1. The amplification products were purified using the MicroCLEAN kit (Microzone, Haywards Heath, UK) and sent to MACROGEN (Madrid, Spain) for Sanger sequencing. The sequences were edited and aligned using MEGA X (Kumar et al. 2018) with the ClustalX method. All sequences have been deposited in GenBank under the following accession numbers: M. surmuletus (PQ339824 - PQ339921), S. cabrilla (PQ339926 - PQ340023), S. notata (PQ363178 - PQ363270), and O. vulgaris (PQ340025 - PQ340124). Genetic diversity and differentiation For data analysis, samples from each species were grouped into three zones according to the sampling sites: FPZ, SCI, and ADJ. For each zone, genetic diversity statistics were estimated using DnaSP v6 (Rozas et al. 2017) . Non‑parametric Kruskal-Wallis and post hoc Dunn’s multiple comparison tests were performed to test statistical differences between zones for each species using the R package FSA (Ogle et al. 2023). The 95% confidence intervals (CIs) for the mean nucleotide diversity values were obtained by 1,000 bootstrap replicates of COI sequences for zone and species, using the pegas R package. Violin plots were generated with ggplot2 package in R (Wickham 2016) to visualize the distribution of nucleotide diversity values of bootstrapped samples across zones. For each species, haplotype networks were created using median-joining network analysis implemented in PopArt v1.7 (Leigh and Bryant 2015). Moreover, genetic differentiation and its statistical significance among the three zones for each species were calculated through pairwise estimates (Φ st ) in Arlequin v3.5 (Excoffier and Lischer 2010), with 10,000 permutations. Demographic history To investigate the demographic history of the studied species in the Menorca Channel, on the one hand, Tajima’s D, Fu’s Fs, and Ramos-Onsins and Rozas's R 2 neutralitystatistics (Tajima 1989; Fu 1997; Ramos-Onsins and Rozas 2002) were calculated with DnaSP v6 (Rozas et al. 2017), with statistical significance assessed through 10,000 permutations. The same software was used to estimate mismatch distributions by comparing the observed distribution of pairwise nucleotide differences with the expected distribution under a model of exponential demographic expansion. To assess how well our experimental data matched the model distribution, the sum of squared deviations (SSD) between the observed and expected mismatch distributions, as well as the raggedness index (rg; Watterson 1975), were used as test statistics. These statistics were performed using 10,000 bootstrapped replicates in Arlequin v3.5 (Excoffier and Lischer 2010). For these analyses, a single population from the Menorca Channel was considered for each species by pooling individuals from the three zones, as long as no clear genetic differentiation was found among them. On the other hand, coalescent simulation analyses using msms program (Ewing and Hermisson 2010) were performed to assess the demographic effects of fisheries protection in the Menorca Channel. Based on the Wright-Fisher neutral model (Wright 1931), the simulations modelled two demographic scenarios starting eight years ago, coinciding with the FPZ declaration: a potential demographic bottleneck in non-protected zones (SCI and ADJ considered separately) with respect to the FPZ, and a potential population expansion in the FPZ with respect to non-fishing protected zones. The simulations assumed that genetic diversity before the establishment of the protected FPZ zone was similar to that in non-protected areas. The simulations assumed no recombination, no migration, and no subsequent demographic events following the initial population size change. The composite parameter θ = 4Neu was approximated by the Watterson’s estimator ( θ W ; Watterson 1975) calculated from the number of segregating sites (S) in DnaSP v6 as theta ( θ ) per COI sequence and sampling zone, as follows: i) For modelling bottlenecks, the observed θ was approximated by the current θ W in the Fisheries Protection Zone (FPZ): θ F = 0.00452 for M. surmuletus ; θ F = 0.00805 for S. cabrilla ; θ F = 0.00708 for S. notata ; and θ F = 0.00346 for O. vulgaris. Bottleneck intensity was calculated as the ratio of nucleotide diversity in individuals from SCI or ADJ ( π S and π A, respectively) to that in individuals from FPZs ( π F ). ii)For modelling expansion, the observed θ was approximated by the average θ W in the non-protected zones SCI and ADJ: θ S-A = 0.00330 for M. surmuletus ; θ S-A = 0.00768 for S. cabrilla ; θ S-A = 0.00504 for S. notata ; and θ S-A = 0.00351 for O. vulgaris . Expansion intensity was estimated as the ratio of nucleotide diversity in FPZ individuals ( π F ) to the average nucleotide diversity in non-protected areas ( π S-A ). To assess changes in genetic diversity, each simulation included 1,000 random samplings of N individuals per species, where N equals the analysed sample size of each species. Results Fishing footprint The spatial distribution of fishing effort in the studied zones showed no evidence of fishing activity within the Fisheries Protection Zone (FPZ) from 2017 to 2023, in contrast to the period from 2009 to 2015 (Fig. 2a and b), confirming the full enforcement of the FPZ. Vessel Monitoring by satellite System (VMS) data also revealed an increase in fishing effort outside the FPZ, especially in southern SCI (Fig. 2c), where bottom trawling activity is now concentrated. In contrast, in the ADJ zone, the intensity of trawling activity remained nearly constant after the FPZ implementation compared to the 2009–2015 period, although fishing effort remained high. Genetic diversity and differentiation Genetic diversity statistics from the COI fragment for each species within each studied zone are presented in Table 1, and the 95% CIs of the mean nucleotide diversity are shown in Table S2. Considering the overall estimates across all zones, Scorpaena notata exhibited the highest number of segregating sites and haplotypes (35 and 32, respectively), whereas Octopus vulgaris presented the lowest values (14 and 11, respectively). On the other hand, Serranus cabrilla showed the highest values across haplotype diversity (Hd = 0.8502), mean number of nucleotide differences (Kt = 5.5620), and nucleotide diversity (π = 0.0091), while Mullus surmuletus (Hd = 0.6933; Kt = 1.5464; π = 0.0026) showed the lowest values. In general, the individuals of the studied species collected from the FPZ displayed the highest nucleotide diversity values compared with those from the non-protected zones, the closest SCI and the farthest ADJ (Table 1, Fig. 3). The lowest levels of nucleotide diversity were observed in the SCI in all three teleost fishes, whereas in O. vulgaris , the lowest value was found in the ADJ. According to the results of the non-parametric Kruskal-Wallis and Dunn’s post hoc tests (Table S3), significant differences were observed between the FPZ and the other two zones for all four species, except between FPZ and ADJ in M. surmuletus. Haplotype networks (Fig. 4) and pairwise estimates (Φ st ; Table S4) for the studied species revealed no population differentiation based on geographic origin within the Menorca Channel, with the main haplotypes shared by individuals from all zones, as expected given the proximity of the areas. Scorpaena notata showed a star-like network, with nearly half of the samples (47%) across all three zones—SCI, FPZ and ADJ—sharing the main haplotype. The other species exhibited two or three main haplotypes. Among them, S. cabrilla presented the most diverse network, with the main haplotype (Hap_2) separated from haplotype Hap_1 by 14 mutational steps. The greatest number of unique haplotypes were found in M. surmuletus and S. notata (both with 72% of their haplotype being unique). Demographic history Negative Tajima’s D and Fu’s Fs values, along with low and significant Ramos-Onsins and Rozas's R 2 values, indicate a demographic expansion in M. surmuletus and S. notata in the Menorca Channel population (Table 2). In contrast, non-significant values were found for S. cabrilla and O. vulgaris populations. Although S. cabrilla showed slightly negative values and O. vulgaris slightly positive values, both values were close to zero, suggesting demographic stability in both species. The unimodal mismatch distributions observed for M. surmuletus and S. notata matched the expected distributions under a sudden expansion model, supporting the neutrality test results. Conversely, S. cabrilla and O. vulgaris exhibited bimodal mismatch distributions (Fig. 5), strongly deviating from the expansion model. From the coalescent simulation tests of bottleneck scenario, the observed nucleotide diversity at the non-protected SCI and ADJ zones for M. surmuletus and S. notata was lower than the expected 95% upper CI limit of nucleotide diversity replicates generated from the simulation (Table 3). This suggests that the proposed model of population bottlenecks caused by fishing in the SCI and ADJ could explain the observed nucleotide diversity estimates. In contrast, the observed nucleotide diversity values for S. cabrilla and O. vulgaris from the SCI and ADJ zones, although lower than in FPZ, did not fall within the 95% upper CI limit of nucleotide diversity estimates expected by the bottleneck model, suggesting no significant effect of the current fishing activities on the genetic diversity levels of these two species. According to the hypothesis of population expansion after banning trawling activities in the protected zone (FPZ), the observed nucleotide diversity values were significantly higher than the expected values from a population expansion derived from the coalescent simulations for all species (Table 3). This could suggest that protection may have allowed genetic diversity recovery to levels higher than expected by our tested model. It could be because the nucleotide diversity in FPZ before protection was higher than the current diversity observed in the other two zones, or because growing rate is higher than in the model. Discussion In the present study, we evaluated how a Fisheries Protection Zone in the Menorca Channel influences genetic diversity by analysing the mitochondrial COI gene in four exploited marine species, three teleost fishes ( Mullus surmuletus , Serranus cabrilla, and Scorpaena notata ) and one cephalopod ( Octopus vulgaris ). Overall, individuals collected from the FPZ displayed the highest nucleotide diversity values compared to those from non-protected zones, the closest SCI and the farthest ADJ. These results suggest a better conservation status for the studied species within the protected area. Recent studies have reported positive effects from the establishment of the FPZ in the Menorca Channel, demonstrating that the 2016 trawling ban has helped protect 80% of rhodolith beds and 95% of coralligenous bottoms from trawling. This protection has led to increased biomass of rhodolith beds and Laminaria rodriguezii forests, thereby contributing to the recovery of epibenthic communities (Farriols et al. 2021; 2025). The distribution of these habitats has increased by 6% and 54%, respectively, in recent years, highlighting the FPZ as an effective conservation measure for benthic habitats, communities, and species in the Menorca Channel (Farriols et al. 2025). The recovery of these significant marine habitats is enhancing marine biodiversity in the area due to their structural and functional complexity (Barberá et al. 2012; Joher et al. 2012, 2015), which benefits both exploited and unexploited species. In addition to the implementation of the FPZ, some areas of the Menorca Channel had already undergone previous changes. On the one hand, submarine cables were installed in the 1970s, in a narrow area within what is now the FPZ, connecting Mallorca and Menorca (Cabrito et al. 2024), which resulted in trawling being excluded from this area for decades, potentially providing long-term benefits to marine communities. On the other hand, a general reduction in fishing effort has been recorded across the Balearic Islands in recent decades (Quetglas et al. 2017). Therefore, the recovery trend observed in this study may have originated from these prior changes and was further enhanced by the implementation of the protected area. Specimens of the four studied species collected from non-protected areas in the Menorca Channel showed high nucleotide diversity values, although lower compared to those from the protected area. This is to be expected given the geographical proximity to the FPZ, suggesting that adjacent areas may benefit from population growth in the protected zone. It is therefore a clear example of how a marine reserve can play a crucial role in sustaining the productivity of fishing zones and contributing to the consequent gene flow between protected and near non-protected areas, which is essential for preserving and restoring ecological processes and for ensuring genetic diversity, which supports species’ ability to adapt to global environmental changes (Gandra et al. 2020). In this sense, the lack of genetic structuring observed in the haplotype networks (Fig. 4) and the low and non-significant Φ st values (Table S4) suggest high levels of gene flow between the FPZ and nearby zones. In the Mediterranean, similar patterns have been observed in fish species such as Diplodus vulgaris , Mullus surmuletus and M. barbatus , where high genetic diversity was found across both protected and adjacent fished areas (Félix-Hackradt et al. 2013; Sahyoun et al. 2016). However, for the three teleost fish species analysed in this study, the lowest nucleotide diversity values were unexpectedly observed in the SCI, the non-protected zone closest to the protected FPZ. As this zone presents similar habitat characteristics to those of protected area (Farriols et al. 2025), the observed reduction in genetic diversity could be attributable to increased fishing pressure rather than habitat differences. Supporting this, Vessel Monitoring System (VMS) data indicate that, following the establishment of the FPZ, trawling activities in the Menorca Channel shifted mainly toward the southern area of the SCI (Fig. 2). While all target species in this study exhibited higher genetic diversity values within the protected area, interspecific differences in nucleotide diversity levels were observed. Specifically, S. cabrilla showed the highest nucleotide diversity, followed by O. vulgaris , whereas M. surmuletus and S. notata presented the lowest values. These differences could be related to differences in the life-history traits of the species studied. A key distinction is S. cabrilla ’s reproductive strategy asa simultaneous hermaphrodite that does not self-fertilize, which reduces homozygosity and inbreeding risk, thereby enhancing genetic diversity by promoting spawning events and offspring heterozygosity (Avise and Mank 2009; Coscia et al. 2016). Since each individual in the population contributes mitochondrial DNA to the next generation—unlike in non-hermaphroditic species, where only females do—the resulting nucleotide diversity is not directly comparable to that of the other species. Regarding O. vulgaris, this species exhibits a fast growth rate, a short semelparous life cycle that allows rapid generational turnover, and high fecundity (Hanlon and Messenger 1996), traits that confer relative resilience to fishing pressure and environmental perturbations compared with many teleost species (Faure et al. 2000) . Lastly, M. surmuletus and S. notata possess typical life-history traits of marine fish: they are oviparous species with external fertilization, their eggs are embedded in a gelatinous matrix, and they have relatively short generation times—3.8 and 2.9 years, respectively (Table 4). In order to compare the nucleotide diversity estimates of the studied species in the Menorca Channel with those reported for other species across the Balearic Islands, only two studies were found (Petit-Marty et al. 2022; Riera et al. 2025), both focusing on teleost species (Fig. 6). Compared with the estimate reported by Petit-Marty et al. (2022) (π = 0.0047), M. surmuletus exhibited lower nucleotide diversity in this study (π = 0.0026), whereas S. notata presented slightly greater diversity (π = 0.0025) than the estimate for the entire population across the Balearic Archipelago (π = 0.0020; Riera et al. 2025). These intraspecific differences may reflect variations in sampling effort and geographic coverage across the Balearic Islands. Additionally, M. surmuletus displayed lower nucleotide estimates than its sister species, M. barbatus (π = 0.0035) (Fig. 6). Both M. surmuletus and S. notata from the Menorca Channel exhibited nucleotide diversity values below the lower limit of the 95% confidence interval for Mediterranean fish species as calculated by Petit-Marty et al. (2022) (mean 95% CI = 0.0034–0.0047) (Fig. 6). In this context, both species exhibited genetic diversity levels slightly higher than two overexploited species, M. merluccius (π = 0.0018) and Lophius budegassa (π = 0.0016) (Fig. 6), both characterized by large body size and long generation times (~10 years) (Petit-Marty et al. 2022). Although the comparison should be interpreted with caution, as the mutation rate may vary among genera, these results suggest that both species could be affected by the impact of fishing. In the case of S. notata , this aligns with the findings of Riera et al. (2025), who reported signs of fishing pressure on this species in the Balearic Islands, despite it being a bycatch species. Finally, S. cabrilla exhibited the highest values even exceeding the upper limit of the 95% confidence interval for Mediterranean fish species (Petit-Marty et al. 2022) (Fig. 6), as expected given its reproduction strategy described previously. The genetic diversity values estimated in the four target species also varied when compared with those of the same species from other areas across the Mediterranean Sea and Northeast Atlantic. M. surmuletus from the Menorca Channel (π = 0.0026) exhibited similar values to populations in Egypt (π = 0.0030) (Soliman et al. 2024). S. cabrilla showed nucleotide diversity values consistent with those reported in the Eastern Mediterranean (π = 0.0118), although populations from Turkey exhibited lower genetic diversity (π = 0.0035) (Bos et al. 2020). Lastly, O. vulgaris exhibited nucleotide diversity levels (π = 0.0045) consistent with estimates from Central Mediterranean (Sardinia, π = 0.0046) (Melis et al. 2018) and Northeast Atlantic populations (Portugal, π = 0.0040; and Canary Islands, π = 0.0059) (Pérez et al. 2023). Regarding demographic history and coalescent simulations analyses, the target species displayed distinct patterns. Populations of M. surmuletus and S. notata in the Menorca Channel showed signs of recent expansion, as indicated by significant neutrality statistics and “star” or “complex”-star-shaped haplotype networks (Table 2, Fig. 4). Similar results have been observed in S. notata population across the Balearic Islands (Riera et al. 2025). In contrast, S. cabrilla and O. vulgaris presented non-significant neutrality statistics and bimodal mismatch distributions, suggesting a stable population size. Bimodal distributions may also indicate complex population structures, such as the presence of distinct lineages (Jenkins et al. 2018), although this was not clearly evident in the haplotype networks. For octopus, two different mitochondrial haplotype lineages have been observed along the East Atlantic coast and locally along Sardinia’s coast (Melis et al. 2018; Quinteiro et al. 2020), as well as a complex genetic pattern across the Mediterranean (Fadhlaoui-Zid et al. 2021). Further analyses should be realised to explore the genetic structure and demographic dynamics of octopus and comber species in the Balearic Islands. Coalescent simulations help assess whether observed results align with expected demographic events such as expansions or bottlenecks in the different zones studied within the Menorca Channel. Coalescent simulations indicated that FPZ populations grew more than expected by a population expansion if the levels of nucleotide diversity in the FPZ populations before protection were equal to those observed in the non-protected zones. The contrary was also true, bottleneck simulations showed that the observed values of nucleotide diversity in the fished zones were significantly lower than expected by a reduction of population starting with the levels observed in the FPZ population. Differences in genetic diversity between ADJ and SCI may stem from historically low diversity levels in the Menorca Channel prior to FPZ implementation. Before protection, high gene flow combined with fishing pressure likely homogenized genetic diversity across zones. However, the establishment of the FPZ enabled population growth within protected areas. In contrast, individuals outside the FPZ faced intense fishing pressure, leading to population decline. The impact was stronger in the closest SCI zone where trawling activities were concentrated (Fig. 2) likely leading to greater genetic erosion than in the farthest ADJ zone, where fishing efforts remained relatively stable following the establishment of the trawling ban. In this scenario, the fishing-driven decline in SCI may have exceeded the demographic benefits of gene flow from the FPZ, whereas the ADJ—although also affected—experienced milder losses. In contrast, for S. cabrilla and O. vulgaris , coalescent simulations also suggested population expansion within the FPZ, but no decline in non-protected zones under the tested conditions. This pattern may result from a) a bottleneck less severe than modelled, b) a recent demographic decline not yet detectable through genetic signals, or c) no significant effect of current trawling activities on the genetic diversity of these species. Additionally, the relatively resilient life history traits of S. cabrilla and O. vulgaris —such as hermaphroditism in the former and rapid turnover in the latter—may have caused these species to recover more quickly from the effects of fishing. These characteristics likely contributed to the higher genetic diversity observed in the present study, making these two species good indicators of conservation effectiveness in both protected and non-protected zones. This study provides genetic evidence of the positive impact of a Fisheries Protection Zone (FPZ) on overexploited marine species in the Menorca Channel, employing mitochondrial COI nucleotide diversity as a novel proxy for species conservation status. Although mitochondrial DNA has several limitations due to its characteristics—maternally inhered, haploidy and lack of recombination—it remains a valuable marker for genetic conservation studies. A positive correlation between mitochondrial and nuclear genetic diversity in fish species has been reported (Piganeau and Eyre-Walker 2009), suggesting that a reduction in mitochondrial COI diversity may reflect broader nuclear genomic erosion and, therefore, can provide useful insight into the conservation status of marine species populations. Overall, our findings indicate a positive trend in the recovery of genetic diversity in exploited species, enhanced by the establishment of the Fisheries Protection Zone, which has contributed to the improved condition of marine habitats within the study area—improvements that are closely linked to the conservation status of marine species. Moreover, this study highlights the value of genetic diversity approaches as effective diagnostic tools to assess protected and non-protected zones, and to support ongoing monitoring of vulnerable and exploited species. Declarations Funding This study is part of the MARFISH project (PDR2020/69–ITS 2017-006), included in the Projectes de Recerca Científica i Tecnològica of the Direcció General de Política Universitària i Recerca , which are funded by the Comunitat Autònoma de les Illes Balears (CAIB) through the Direcció Conselleria de Fons Europeus, Universitat i Cultura, with resources from the Tourist Stay Tax Law. The CAIB also funded the 2022 grants for the training of research personnel (N. Pasini, research grant nº FPI_026_2022). The MEDITS surveys are cofunded by the European Union through the European Maritime Fisheries and Aquaculture Fund (EMFAF) within the National Program of collection, management, and use of data in the fisheries sector and support for scientific advice regarding the Common Fisheries Policy. The CANAL surveys were part of the SosMed project (Improvement in scientific and technical knowledge for the sustainability of demersal fisheries in the western Mediterranean), funded by the European Union—Next Generation (Recovery, Transformation and Resilience Plan), with an agreement between the Ministry of Agriculture, Fisheries and Food and the Spanish National Research Council, through the Spanish Institute of Oceanography. Conflicts of interest The authors declare no conflict of interest. Author Contributions Conceptualization, F.O., A.P. and S.R.-A.; methodology, N.P.; data analysis, N.P., S.R.-A., M.B., J.F.F, M.T.F. and N.P.-M.; writing—original draft preparation, N.P.; writing—review and editing, M.B., J.F.F., M.T.F., N.P.-M., F.O., S.R.-A. and A.P.; funding acquisition, F.O. and A.P. All authors have read and agreed to the published version of the manuscript. Data availability All data supporting the findings of this study are available within the paper and its Supplementary Information. COI mitochondrial sequences were deposited into the GenBank (NCBI) database under accession numbers: PQ339824-PQ339921 ( Mullus surmuletus ); PQ339926-PQ340023 ( Serranus cabrilla ); PQ363178-PQ363270 ( Scorpaena notata ); and PQ340025-PQ340124 ( Octopus vulgaris ). References Allcock L, Headlam J, Allen G (2018) Octopus vulgaris. 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Symbols are defined in the footnote a ZONE N S h Hd ± SD Kt π ± SD Mullus surmuletus FPZ 33 11 9 0.6987 ± 0.0740 1.6894 0.0028 ± 0.0004 SCI 34 7 7 0.6292 ± 0.0630 1.2674 0.0021 ± 0.0002 ADJ 31 9 8 0.7505 ± 0.0500 1.6430 0.0027 ± 0.0003 Total 98 20 18 0.6933 ± 0.0360 1.5464 0.0026 ± 0.0002 Serranus cabrilla FPZ 33 19 11 0.8712 ± 0.0290 5.8371 0.0095 ± 0.0006 SCI 34 17 10 0.7825 ± 0.0630 5.3957 0.0088 ± 0.0009 ADJ 31 19 12 0.8645 ± 0.0390 5.5828 0.0091 ± 0.0006 Total 98 28 21 0.8502 ± 0.0200 5.5620 0.0091 ± 0.0004 Scorpaena notata FPZ 30 18 13 0.7954 ± 0.0680 2.2643 0.0035 ± 0.0007 SCI 31 9 10 0.6237 ± 0.0990 0.8086 0.0013 ± 0.0003 ADJ 32 17 16 0.8528 ± 0.0570 1.5806 0.0025 ± 0.0004 Total 93 35 32 0.7632 ± 0.0460 1.5680 0.0024± 0.0003 Octopus vulgaris FPZ 30 9 6 0.7770 ± 0.0380 3.5012 0.0053 ± 0.0004 SCI 34 10 7 0.7683 ± 0.0450 3.4029 0.0052 ± 0.0003 ADJ 36 9 6 0.7492 ± 0.0460 3.3397 0.0051 ± 0.0005 Total 100 14 11 0.7713 ± 0.0180 3.3851 0.0052 ± 0.0002 a N = number of individuals; S = number of segregating sites; h = number of haplotypes; Hd = haplotype diversity; Kt = mean nucleotide difference; π = nucleotide diversity; SD = standard deviation Table 2 Tajima’s D, Fu’s FS, Ramos-Onsins and Rozas's R 2 indices, as well as mismatch distribution metrics (including the sum of squared deviations from the sudden expansion model, SSD, and the Harpending raggedness index, rg), are indicated for each species. The corresponding p-values are reported in brackets. Values in bold were significant at the 95% confidence level Species D Fs R 2 SSD rg Mullus surmuletus -1.7452 ( 0.0145 ) -10.6423 ( 0.0002 ) 0.0371 ( 0.0391 ) 0.0261 (0.2509) 0.0883 (0.1621) Serranus cabrilla -0.0332 (0.6000) -1.9033 (0.3163) 0.0958 (0.5727) 0.0307 (0.2609) 0.0333 (0.4172) Scorpaena notata -2.4020 ( 0.0000 ) -28.0227 ( 0.0000 ) 0.0210 ( 0.0000 ) 0.0134 (0.2220) 0.0758 (0.2260) Octopus vulgaris 0.6886 (0.8018) 0.8488 (0.6770) 0.1173 (0.7849) 0.0786 (0.0537) 0.1050 (0.1579) Table 3 Observed (π ob ) and expected (π ex ) nucleotide diversity values are shown for each species in the three zones: the Fishery Protection Zone (FPZ), where trawling is banned; the Site of Community Importance (SCI), the nearest open-to-fishing zone to the FPZ; and the ADJ zone, the farthest open-to-fishing area from the FPZ. Observed values were calculated using DnaSP v6. Expected values were modelled through coalescent simulations assuming equal genetic diversity across zones prior to the FPZ’s establishment. Each simulation used θ ( θ = 4Neu) as starting parameter, estimated by the observed Watterson’s estimator ( θ W ) in FPZ samples (for bottleneck scenarios) and by the observed average θ W in the non-protected zones SCI and ADJ specimens (for expansion scenarios), with 1,000 random samplings of N individuals per species ( N = observed sample size). Bottleneck intensity was calculated as the ratio of nucleotide diversity in individuals from SCI or ADJ (π S and π A , respectively) to those from FPZs (π F ); expansion intensity was calculated as the ratio of π F to the average nucleotide diversity in non-protected areas (π S-A ). The 95% upper CI limit of the 1,000 nucleotide diversity replicates obtained from the msms program are indicated Bottleneck simulations Mullus surmuletus Serranus cabrilla Bottleneck intensity π ob π ex 95% upper CI limit Bottleneck intensity π ob π ex 95% upper CI limit SCI 0.74822 0.00211 0.00332 0.00345 0.95597 0.00882 0.00725 0.00750 ADJ 0.97163 0.00274 0.00427 0.00443 0.92453 0.00912 0.00766 0.00794 Scorpaena notata Octopus vulgaris Bottleneck intensity π ob π ex 95% upper CI limit Bottleneck intensity π ob π ex 95% upper CI limit SCI 0.69688 0.00126 0.00270 0.00282 0.95310 0.00518 0.00348 0.00361 ADJ 0.35694 0.00246 0.00482 0.00499 0.97186 0.00508 0.00339 0.00353 Population expansion simulations Mullus surmuletus Serranus cabrilla Expansion intensity π ob π ex 95% upper CI limit Expansion intensity π ob π ex 95% upper CI limit FPZ 1.16289 0.00282 0.00232 0.00240 1.06354 0.00954 0.00535 0.00550 Scorpaena notata Octopus vulgaris Expansion intensity π ob π ex 95% upper CI limit Expansion intensity π ob π ex 95% upper CI limit FPZ 1.89785 0.00353 0.00303 0.00310 1.03899 0.00533 0.00243 0.00250 Table 4 Characteristics of the studied species. Max. size and gen. times define the maximum recorded size in centimetres and the mean generation time in years, respectively. Information on commercial importance, fishing gear, IUCN status, and movement/habitats was obtained from IUCN reports for the evaluated species and their populations in the Mediterranean. Data on maximum size and generation times were retrieved from FishBase. N/A= not available Species Commercial importance Fishing gear IUCN Population trends Threats Movements/Habitats Max. size (cm)/gen. times (year) Mullus surmuletus High (Carpenter et al. 2015) Bottoms trawls, trammel nets (Carpenter et al. 2015) LC (Carpenter et al. 2015) Decreasing (Quetglas et al. 2016) Overexploitation (FAO 2024) Oceanodromous/ Marine oceanic - Epipelagic (0-200 m) (Carpenter et al. 2015) 40.0 cm/3.8 years Serranus cabrilla Minor (Smith-Vaniz and Iwamoto 2015) Recreational fishing; bycatch in gillnet, bottom longline and trawls (Smith-Vaniz and Iwamoto 2015) LC (Smith-Vaniz and Iwamoto 2015) Stable (Smith-Vaniz and Iwamoto 2015) Overexploitation (Quetglas et al. 2016) Low migrant/ Marine neritic - Subtidal rock, sandy-mud (Smith-Vaniz and Iwamoto 2015) 40.0 cm/5.7 years Scorpaena notata Minor (Nunoo et al. 2015) Bycatch in trammel net and bottom trawls (Nunoo et al. 2015) LC (Nunoo et al. 2015) N/A N/A Low migrant/ Marine neritic – deep benthic (Nunoo et al. 2015) 26.0 cm/2.9 years Octopus vulgaris High (Allcock et al. 2018) Traps and spots, trammel nets, bottom trawls (Allcock et al. 2018) LC (Allcock et al. 2018) N/A Overexploitation (FAO 2024) Migrant/ Marine neritic - Subtidal rock, sandy-mud, macroalgal/kelp (Allcock et al. 2018) 180 cm/1.2 years Additional Declarations No competing interests reported. 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07:09:17","extension":"png","order_by":32,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":72221,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/6c8505f73807a42df7c87ed4.png"},{"id":95857509,"identity":"05d14355-420b-43f6-9484-b09f8912201d","added_by":"auto","created_at":"2025-11-13 17:01:39","extension":"png","order_by":33,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":104603,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/86d5b0e9ca096484e73659b8.png"},{"id":96241758,"identity":"5d61902a-4fd0-4295-be27-0aeb78abc103","added_by":"auto","created_at":"2025-11-19 07:11:19","extension":"png","order_by":34,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":37964,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/483e77d0d82c9f38c46d4561.png"},{"id":96240904,"identity":"2d61e644-9c28-46c7-895f-0b159f556a2e","added_by":"auto","created_at":"2025-11-19 07:09:39","extension":"xml","order_by":35,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":252724,"visible":true,"origin":"","legend":"","description":"","filename":"4bce194ca48f4cd69918cbf0a5c0fb331structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/a4b2e7944ffd97f8d71e757f.xml"},{"id":95857510,"identity":"adcc7bef-ace5-414b-a702-e15dfc4e826b","added_by":"auto","created_at":"2025-11-13 17:01:39","extension":"html","order_by":36,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":267370,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/5a019198a7c47a6464d2f92b.html"},{"id":95857468,"identity":"f8f2eb69-3f73-430d-8cc2-6d7bd6ab7434","added_by":"auto","created_at":"2025-11-13 17:01:38","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":3776045,"visible":true,"origin":"","legend":"\u003cp\u003eSampling sites where specimens of the studied species were collected. Each species is represented by a different symbol: circles for \u003cem\u003eMullus surmuletus\u003c/em\u003e, squares for \u003cem\u003eSerranus cabrilla\u003c/em\u003e, triangles for \u003cem\u003eScorpaena notata\u003c/em\u003e, and diamonds for \u003cem\u003eOctopus vulgaris\u003c/em\u003e. Colours indicate the studied zones: the Fisheries Protection Zone (FPZ), where trawling is banned, is shown in red; the Site of Community Importance (SCI), the nearest open-to-fishing zone to the FPZ, is shown in light blue; and the ADJ zone, the farthest open-to-fishing area from the FPZ, is shown in green\u003c/p\u003e","description":"","filename":"Figure1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/a2b9665e2b61d80744e2748b.jpeg"},{"id":95857469,"identity":"b25e82b1-cf98-4066-a753-8752a447a087","added_by":"auto","created_at":"2025-11-13 17:01:38","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":987938,"visible":true,"origin":"","legend":"\u003cp\u003eMean annual number of Vessel Monitoring by satellite System (VMS) signals from fishing vessels throughout the Menorca Channel: a) during the period 2009-2015, and b) during 2017-2023, representing the periods before and after the Fisheries Protection Zone (FPZ) declaration, respectively. c) Differences in the mean annual number of VMS signals between the two periods (2017-2023 vs. 2009-2015). Colours on the map indicate different levels of the mean annual number of VMS signals. In panel (c), green indicates a reduction in VMS signals following the FPZ declaration, while red indicates an increase\u003c/p\u003e","description":"","filename":"Figure2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/74e63cf9abc887a9a1558d73.jpeg"},{"id":96240911,"identity":"2fd4cc81-c63d-4178-a2fd-eca65501b3d8","added_by":"auto","created_at":"2025-11-19 07:09:41","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":597294,"visible":true,"origin":"","legend":"\u003cp\u003eViolin plots representing the distribution of nucleotide diversity values among bootstrapped samples for each species in the three studied zones. The Fisheries Protection Zone (FPZ), where trawling is banned, is shown in red. The Site of Community Importance (SCI), the nearest open-to-fishing zone to the FPZ, is shown in light blue. The ADJ zone, the farthest open-to-fishing area from the FPZ, is shown in green. Red dots indicate the mean values, while black dots represent outliers. The boxes within the plots show the interquartile range\u003c/p\u003e","description":"","filename":"Figure3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/ba389317c498d33ad7910980.jpeg"},{"id":96241139,"identity":"ff21bd87-5570-46ab-872b-f100771d5de5","added_by":"auto","created_at":"2025-11-19 07:10:16","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":380610,"visible":true,"origin":"","legend":"\u003cp\u003eMedian-joining haplotype networks of mitochondrial COI sequences for each species: a) \u003cem\u003eMullus surmuletus\u003c/em\u003e; b) \u003cem\u003eSerranus cabrilla\u003c/em\u003e; c) \u003cem\u003eScorpaena notata\u003c/em\u003e; d) \u003cem\u003eOctopus vulgaris\u003c/em\u003e. Size of circles is proportional to haplotype frequencies, orthogonal bars between branch nodes indicate single mutational steps and black nodes represent unsampled intermediate haplotypes\u003c/p\u003e","description":"","filename":"Figure4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/f519cafe19d233dfe2b75a34.jpeg"},{"id":95857481,"identity":"6f4f4cbc-d9c9-4800-8925-588bc1683bd6","added_by":"auto","created_at":"2025-11-13 17:01:38","extension":"jpeg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":648819,"visible":true,"origin":"","legend":"\u003cp\u003eObserved (blue) and expected (orange) mismatch distributions under the sudden expansion model for the populations of the four target species in the Menorca Channel\u003c/p\u003e","description":"","filename":"Figure5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/69c8c0d11987bcc0f2924ed9.jpeg"},{"id":96241222,"identity":"96e3443e-4627-4931-87a4-ac1c2336fc1c","added_by":"auto","created_at":"2025-11-19 07:10:24","extension":"jpeg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":252096,"visible":true,"origin":"","legend":"\u003cp\u003eLevels of nucleotide diversity in the COI mitochondrial gene for the studied species and other species from the Balearic Islands. Blue icons represent populations from the Menorca Channel, green icons indicate nucleotide diversity estimates for four Mediterranean species in the Balearic Islands reported by Petit-Marty et al. (2022), while orange icons represent the nucleotide diversity value for the entire population of \u003cem\u003eScorpaena notata\u003c/em\u003e in the Balearics Islands (Riera et al. 2025). The abbreviations stand for: MS = \u003cem\u003eMullus surmuletus\u003c/em\u003e; MB = \u003cem\u003eMullus barbatus;\u003c/em\u003e SC = \u003cem\u003eSerranus cabrilla\u003c/em\u003e; SN = \u003cem\u003eScorpaena notata\u003c/em\u003e; LB = \u003cem\u003eLophius budegassa\u003c/em\u003e; MM = \u003cem\u003eMerluccius merluccius\u003c/em\u003e. The red and green lines represent, respectively, the lower and upper boundaries of the 95% CI of the mean distribution of bootstrapped nucleotide diversity estimates for Mediterranean fish species, as calculated by Petit-Marty et al. (2022)\u003c/p\u003e","description":"","filename":"Figure6.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/ce1f2cca741680c16b012522.jpeg"},{"id":96255126,"identity":"81440185-2cfb-4851-b2ba-8eb683b96fa0","added_by":"auto","created_at":"2025-11-19 07:47:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":7903174,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/93bd2fa1-9d10-4873-9b70-a94ad7b34643.pdf"},{"id":95857475,"identity":"dfe63a8e-4daf-4d64-973b-080fd91d47bc","added_by":"auto","created_at":"2025-11-13 17:01:38","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":258911,"visible":true,"origin":"","legend":"","description":"","filename":"SIGeneticdiversityPasinietal.2025.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7271616/v1/a5483bd0177c5c53db4cbb67.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Genetic diversity in mitochondrial DNA reveals the effect of a Fisheries Protection Zone on exploited marine species in the Menorca Channel (Western Mediterranean)","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIn the last few decades, the implementation of marine protected areas (MPAs) and fisheries protection zones (FPZs) has increased worldwide, implemented as tools to simultaneously achieve both biodiversity conservation and fisheries management objectives (Hilborn \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Gaines et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Protected areas play an important role in fisheries management due to two main ecological mechanisms: the dispersal of adults and juveniles, defined as \"spill-over\" (Rowley \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e1994\u003c/span\u003e), and the larval supply. These areas contribute to the restoration of overfished fish stocks and provide net benefits to nearby areas (Di Lorenzo et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2016\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Andrello et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In the Mediterranean Sea, a clear increase in biomass and abundance of exploited marine species within protected areas has been observed, with consequent benefits for fishing activities in areas adjacent to the reserves (Guidetti and Sala \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Harmelin-Vivien et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Follesa et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Giakoumi et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). These benefits have also been observed in individual fish conditions, assessed through physiological indicators (Lloret and Planes \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Lloret et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Sillero-R\u0026iacute;os et al. \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). These findings suggest that some fish species show a preference for habitats with better conditions for development, survival, feeding, and reproduction.\u003c/p\u003e\u003cp\u003eFurthermore, it is known that the exploitation of fishery resources can affect the evolutionary dynamics of fish populations in different ways: i) loss of genetic diversity or adaptive potential (Pinsky and Palumbi \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Petit-Marty et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e); ii) variations in the power of detection of population structure (Gandra et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e); and iii) evolutionary changes due to fishing pressures (Allendorf et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Intense fishing pressure leads to population declines, which will significantly reduce the levels of genetic diversity if declines are strong enough to produce changes in the frequency of alleles within the affected populations (Hauser et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Hutchinson et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Ruggeri et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Since genetic variation is the basis of natural selection, its degradation can reduce the ability of a species to adapt to changing environments produced directly or indirectly by anthropogenic pressures, increasing the risk of local extinction (Spielman et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Allendorf et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; V\u0026aacute;squez et al. \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn this sense, overfishing drives population decline, which in turn causes the loss of genetic diversity in exploited species and, therefore, potentially affects their conservation and adaptive capacity to environmental changes (Petit-Marty et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Sadler et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Mendoza-Portillo et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn this context, mitochondrial genes can provide information not only on demographic changes and declines in population sizes but can also serve as diagnostic tools for assessing the conservation status of exploited populations (Johnson et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Petit-Marty et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Righi et al. \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Ilham Syahadah Mohd Yusoff et al. 2021; Canteri et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Ferragut-Perello et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Zlateva et al. \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Recently, some studies have demonstrated that estimates of genetic diversity\u0026mdash;particularly the nucleotide diversity index\u0026mdash;derived from the mitochondrial marker Cytochrome C Oxidase subunit I (COI) can be effective, simple and cost-efficient indicators of the conservation status of commercially exploited (Yorisue et al. \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Petit-Marty et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) and vulnerable (Ham-Due\u0026ntilde;as et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Petit-Marty et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Ferragut-Perello et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) marine fish species.\u003c/p\u003e\u003cp\u003eThe genetic diversity observed in mitochondrial markers, combined with coalescent simulations, also provides insights into the demographic history of marine species over time. Coalescent simulations are a well-established method for generating population samples under different evolutionary scenarios (Ewing and Hermisson \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) and have been used to estimate expected decreases in genetic diversity\u0026mdash;under a neutral model of evolution\u0026mdash;resulting from population declines driven by sustained harvesting pressure (Pinsky and Palumbi \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Arenas \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Petit-Marty et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Reid and Pinsky \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe present study was carried out in the Menorca Channel, which is located between the islands of Mallorca and Menorca in the Western Mediterranean and represents nearly 20% of the coastal continental shelf of the Balearic Archipelago (40 to 100 m depth). Due to the hydrodynamic conditions, the area hosts a wide distribution of habitats and species of conservation interest, such as coralligenous outcrops, ma\u0026euml;rl, and biogenic detrital beds (Barber\u0026aacute; et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Moranta et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Farriols et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Red algae beds, composed of both \u003cem\u003ePeyssonnelia\u003c/em\u003e and rhodoliths, have a high biodiversity and secondary macrobenthic production, which positively influence the abundance, physiological status, and key vital aspects of demersal resources on the Balearic continental shelf (Ordines et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2009\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Ma\u0026euml;rl and coralligenous beds are classified as sensitive habitats (HSs) and essential fish habitats (EFHs) (Ardizzone \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Following the implementation of Council Regulation (EC) No. 1967/2006, which establishes management measures for the sustainable exploitation of fishery resources in the Mediterranean Sea, rhodolith beds and coralligenous bottoms were recognized as protected habitats. Owing to the presence of these essential habitats and their spatial overlap with trawling activities and associated impacts, the Menorca Channel was declared a Site of Community Importance (SCI) in 2014. In 2016, trawling was subsequently banned in certain areas within the SCI, designated as Fisheries Protection Zone (FPZ).\u003c/p\u003e\u003cp\u003eThis study aims to assess the effect of a Fisheries Protection Zone (FPZ) on the conservation status of exploited marine species by analysing genetic diversity levels using the mitochondrial COI marker, comparing the FPZ located in the Menorca Channel with the surrounding areas subjected to fishing activity. To do this, three species of teleosts were used as models: \u003cem\u003eMullus surmuletus\u003c/em\u003e (red striped mullet), \u003cem\u003eSerranus cabrilla\u003c/em\u003e (comber) and \u003cem\u003eScorpaena notata\u003c/em\u003e (small red scorpionfish), and one cephalopod species: \u003cem\u003eOctopus vulgaris\u003c/em\u003e (common octopus). All these species are widely distributed throughout the East Atlantic and the Mediterranean Sea, inhabiting the narrow continental shelf areas characterized by rocky or sandy substrates and seagrass meadows (Ordines et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Al\u0026oacute;s et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Fadhlaoui-Zid et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; F\u0026eacute;lix-Hackradt et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Bos et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; P\u0026eacute;rez et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Furthermore, in the Balearic Islands all species are commercially exploited, whether they are target species (\u003cem\u003eM. surmuletus\u003c/em\u003e, \u003cem\u003eS. cabrilla, and O. vulgaris\u003c/em\u003e) or bycatches (\u003cem\u003eS. notata\u003c/em\u003e) (Quetglas et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003e\u003cstrong\u003eSampling\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe samples of the studied species were collected during the MEDITS (International bottom trawl survey in the Mediterranean) and CANAL scientific surveys, which were partially or totally conducted in the Menorca Channel between 2021 and 2023. In these surveys, the experimental bottom trawl GOC-73 was used to sample demersal communities and resources from seafloor areas subject to fishing activity (for specific information see Spedicato et al. 2019). Specifically, the samples were obtained from FPZs located in the Menorca Channel and from surrounding fished area within the SCI and adjacent areas (ADJ), located south of the Menorca Channel. Sampling sites for the studied species are indicated in Fig. 1. For all the studied species, tissue samples were collected from at least 30 specimens per zone (FPZ, SCI, and ADJ), preserved in 96% ethanol and stored at -20ºC. The total number of tissue samples is indicated in Table 1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFishing footprint\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData from the Vessel Monitoring by satellite System (VMS) collected between 2009 and 2023, were analysed to assess the distribution of bottom trawl fishing efforts along the Menorca Channel before and after the establishment of the FPZ. Prior to analysis, VMS data were filtered to exclude signals produced during sailing and manoeuvring. To do so, only VMS signals recorded between 05:00 am and 05:00 pm (the period when trawlers are allowed to fish in the Balearic Islands) and with an instantaneous speed from 2 to 3.6 knots (the velocity range during fishing operations in the Balearic Islands) were considered. Finally, the mean annual number of VMS signals per cell in a 0.01º resolution grid was mapped with ArcGIS Desktop 10.8 (ESRI 2020) for the periods 2009-2015 and 2017-2023, corresponding to the sampling periods before and after the FPZ implementation, respectively.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLaboratory procedures\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTotal genomic DNA (gDNA) was extracted using the DNeasy Blood and Tissue kit (Qiagen, West Sussex, UK). Polymerase chain reaction (PCR) was used to amplify the genetic marker Cytochrome C Oxidase subunit I (COI) using different primer sets: FF2d/FR1d (600 bp for \u003cem\u003eM. surmuletus\u003c/em\u003e, 642 bp for \u003cem\u003eS. notata\u003c/em\u003e;Ivanova et al. 2007), FishF1/FishR1 (612 bp for \u003cem\u003eS. cabrilla\u003c/em\u003e; Ward et al. 2005), and LCO1490/HCO2198 (657 bp for \u003cem\u003eO. vulgaris\u003c/em\u003e; Folmer et al. 1994). PCR reactions were carried out in a 25 μL volume containing: 2.5 μL of 10X Buffer (BIOLINE, London, UK),\u0026nbsp;1.75 μL of 50 mM MgCl\u003csub\u003e2\u003c/sub\u003e, 1 μL of 100 mM dNTPs, 0.5 μL of each primer (200 mM), 0.05 μL of DNA polymerase (BIOLINE)\u0026nbsp;(5 U/μL), and 1 μL of gDNA template.\u0026nbsp;PCR program conditions for each species and primer set are detailed in Table S1. The amplification products were purified using the MicroCLEAN kit (Microzone, Haywards Heath, UK) and sent to MACROGEN (Madrid, Spain) for Sanger sequencing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe sequences were edited and aligned using MEGA X (Kumar et al. 2018)\u0026nbsp;with the ClustalX method. All sequences have been deposited in GenBank under the following accession numbers: \u003cem\u003eM. surmuletus\u003c/em\u003e (PQ339824 - PQ339921), \u003cem\u003eS. cabrilla\u003c/em\u003e (PQ339926 - PQ340023), \u003cem\u003eS. notata\u003c/em\u003e (PQ363178 - PQ363270), and \u003cem\u003eO. vulgaris\u0026nbsp;\u003c/em\u003e(PQ340025 - PQ340124).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenetic diversity and differentiation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor data analysis, samples from each species were grouped into three zones according to the sampling sites: FPZ, SCI, and ADJ. For each zone, genetic diversity statistics were estimated using DnaSP v6 (Rozas et al. 2017) . Non‑parametric Kruskal-Wallis and \u003cem\u003epost hoc\u003c/em\u003e Dunn’s multiple comparison tests were performed to test statistical differences between zones for each species using the R package \u003cem\u003eFSA\u003c/em\u003e (Ogle et al. 2023). The 95% confidence intervals (CIs) for the mean nucleotide diversity values were obtained by 1,000 bootstrap replicates of COI sequences for zone and species, using the \u003cem\u003epegas\u003c/em\u003e R package. Violin plots were generated with \u003cem\u003eggplot2\u003c/em\u003e package in R (Wickham 2016) to visualize the distribution of nucleotide diversity values of bootstrapped samples across zones.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFor each species, haplotype networks were created using median-joining network analysis implemented in PopArt v1.7 (Leigh and Bryant 2015). Moreover, genetic differentiation and its statistical significance among the three zones for each species were calculated through pairwise estimates (Φ\u003csub\u003est\u003c/sub\u003e) in Arlequin\u0026nbsp;v3.5 (Excoffier and Lischer 2010), with 10,000 permutations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDemographic history\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo investigate the demographic history of the studied species in the Menorca Channel, on the one hand, Tajima’s D, Fu’s Fs, and Ramos-Onsins and Rozas's R\u003csub\u003e2\u0026nbsp;\u003c/sub\u003eneutralitystatistics (Tajima 1989; Fu 1997; Ramos-Onsins and Rozas 2002) were calculated with DnaSP v6 (Rozas et al. 2017), with statistical significance assessed through 10,000 permutations. The same software was used to estimate\u0026nbsp;mismatch distributions by comparing the observed distribution of pairwise nucleotide differences with the expected distribution under a model of exponential demographic expansion. To assess how well our experimental data matched the model distribution, the sum of squared deviations (SSD) between the observed and expected mismatch distributions, as well as the raggedness index (rg; Watterson 1975), were used as test statistics. These statistics were performed using 10,000 bootstrapped replicates in Arlequin v3.5 (Excoffier and Lischer 2010). For these analyses, a single population from the Menorca Channel was considered for each species by pooling individuals from the three zones, as long as no clear genetic differentiation was found among them.\u003c/p\u003e\n\u003cp\u003eOn the other hand, coalescent simulation analyses using \u003cem\u003emsms\u003c/em\u003e program (Ewing and Hermisson 2010) were performed to assess the demographic effects of fisheries protection in the Menorca Channel.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBased on the Wright-Fisher neutral model (Wright 1931), the simulations modelled two demographic scenarios starting eight years ago, coinciding with the FPZ declaration: a potential demographic bottleneck in non-protected zones (SCI and ADJ considered separately) with respect to the FPZ, and a potential population expansion in the FPZ with respect to non-fishing protected zones. The simulations assumed that genetic diversity before the establishment of the protected FPZ zone was similar to that in non-protected areas. The simulations assumed no recombination, no migration, and no subsequent demographic events following the initial population size change. The composite parameter \u003cem\u003eθ\u003c/em\u003e\u003cem\u003e\u0026nbsp;=\u0026nbsp;\u003c/em\u003e4Neu was approximated by the Watterson’s estimator (\u003cem\u003eθ\u003c/em\u003e\u003cem\u003e\u003csub\u003eW\u003c/sub\u003e\u003c/em\u003e; Watterson 1975) calculated from the number of segregating sites (S) in DnaSP v6 as theta (\u003cem\u003eθ\u003c/em\u003e) per COI sequence and sampling zone, as follows:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ei) For modelling bottlenecks, the observed \u003cem\u003eθ\u0026nbsp;\u003c/em\u003ewas approximated by the current \u003cem\u003eθ\u003csub\u003eW\u003c/sub\u003e\u0026nbsp;\u003c/em\u003ein the Fisheries Protection Zone (FPZ): \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eF\u003c/sub\u003e = 0.00452 for \u003cem\u003eM. surmuletus\u003c/em\u003e; \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eF\u003c/sub\u003e = 0.00805 for \u003cem\u003eS. cabrilla\u003c/em\u003e; \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eF\u003c/sub\u003e = 0.00708 for \u003cem\u003eS. notata\u003c/em\u003e; and \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eF\u003c/sub\u003e = 0.00346 for \u003cem\u003eO. vulgaris.\u0026nbsp;\u003c/em\u003eBottleneck intensity was calculated as the ratio of nucleotide diversity in individuals from SCI or ADJ (\u003cem\u003eπ\u003c/em\u003e\u003csub\u003eS\u003c/sub\u003e and \u003cem\u003eπ\u003c/em\u003e\u003csub\u003eA,\u0026nbsp;\u003c/sub\u003erespectively) to that in individuals from FPZs (\u003cem\u003eπ\u003c/em\u003e\u003csub\u003eF\u003c/sub\u003e).\u003c/p\u003e\n\u003cp\u003eii)For modelling expansion, the observed \u003cem\u003eθ\u003c/em\u003ewas approximated by the average \u003cem\u003eθ\u003c/em\u003e\u003cem\u003e\u003csub\u003eW\u003c/sub\u003e\u0026nbsp;\u003c/em\u003ein the non-protected zones SCI and ADJ: \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eS-A\u003c/sub\u003e = 0.00330 for \u003cem\u003eM. surmuletus\u003c/em\u003e; \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eS-A\u003c/sub\u003e = 0.00768 for \u003cem\u003eS. cabrilla\u003c/em\u003e; \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eS-A\u003c/sub\u003e = 0.00504 for \u003cem\u003eS. notata\u003c/em\u003e; and \u003cem\u003eθ\u003c/em\u003e\u003csub\u003eS-A\u003c/sub\u003e = 0.00351 for \u003cem\u003eO. vulgaris\u003c/em\u003e. Expansion intensity was estimated as the ratio of nucleotide diversity in FPZ individuals (\u003cem\u003eπ\u003c/em\u003e\u003csub\u003eF\u003c/sub\u003e) to the average nucleotide diversity in non-protected areas (\u003cem\u003eπ\u003c/em\u003e\u003csub\u003eS-A\u003c/sub\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTo assess changes in genetic diversity, each simulation included 1,000 random samplings of \u003cem\u003eN\u003c/em\u003e individuals per species, where \u003cem\u003eN\u003c/em\u003e equals the analysed sample size of each species.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eFishing footprint\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe spatial distribution of fishing effort in the studied zones showed no evidence of fishing activity within the Fisheries Protection Zone (FPZ) from 2017 to 2023, in contrast to the period from 2009 to 2015 (Fig. 2a and b), confirming the full enforcement of the FPZ. Vessel Monitoring by satellite System (VMS) data also revealed an increase in fishing effort outside the FPZ, especially in southern SCI (Fig. 2c), where bottom trawling activity is now concentrated. In contrast, in the ADJ zone, the intensity of trawling activity remained nearly constant after the FPZ implementation compared to the 2009–2015 period, although fishing effort remained high.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenetic diversity and differentiation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGenetic diversity statistics from the COI fragment for each species within each studied zone are presented in Table 1, and the 95% CIs of the mean nucleotide diversity are shown in Table S2.\u003c/p\u003e\n\u003cp\u003eConsidering the overall estimates across all zones, \u003cem\u003eScorpaena notata\u003c/em\u003e exhibited the highest number of segregating sites and haplotypes (35 and 32, respectively), whereas \u003cem\u003eOctopus vulgaris\u003c/em\u003e presented the lowest values (14 and 11, respectively). On the other hand, \u003cem\u003eSerranus cabrilla\u003c/em\u003e showed the highest values across haplotype diversity (Hd = 0.8502), mean number of nucleotide differences (Kt = 5.5620), and nucleotide diversity (π = 0.0091), while \u003cem\u003eMullus surmuletus\u003c/em\u003e (Hd = 0.6933; Kt = 1.5464; π = 0.0026) showed the lowest values.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn general, the individuals of the studied species collected from the FPZ displayed the highest nucleotide diversity values compared with those from the non-protected zones, the closest SCI and the farthest ADJ (Table 1, Fig. 3). The lowest levels of nucleotide diversity were observed in the SCI in all three teleost fishes, whereas in \u003cem\u003eO. vulgaris\u003c/em\u003e, the lowest value was found in the ADJ. According to the results of the non-parametric Kruskal-Wallis and Dunn’s \u003cem\u003epost hoc\u003c/em\u003e tests (Table S3), significant differences were observed between the FPZ and the other two zones for all four species, except between FPZ and ADJ in \u003cem\u003eM. surmuletus.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHaplotype networks (Fig. 4) and pairwise estimates (Φ\u003csub\u003est\u003c/sub\u003e; Table S4) for the studied species revealed no population differentiation based on geographic origin within the Menorca Channel, with the main haplotypes shared by individuals from all zones, as expected given the proximity of the areas. \u003cem\u003eScorpaena notata\u003c/em\u003e showed a star-like network, with nearly half of the samples (47%) across all three zones—SCI, FPZ and ADJ—sharing the main haplotype. The other species exhibited two or three main haplotypes. Among them, \u003cem\u003eS. cabrilla\u003c/em\u003e presented the most diverse network, with the main haplotype (Hap_2) separated from haplotype Hap_1 by 14 mutational steps. The greatest number of unique haplotypes were found in \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e (both with 72% of their haplotype being unique).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDemographic history\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNegative Tajima’s D and Fu’s Fs values, along with low and significant Ramos-Onsins and Rozas's R\u003csub\u003e2\u003c/sub\u003e values, indicate a demographic expansion in \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e in the Menorca Channel population (Table 2). In contrast, non-significant values were found for \u003cem\u003eS. cabrilla\u003c/em\u003e and \u003cem\u003eO. vulgaris\u003c/em\u003e populations. Although S. cabrilla showed slightly negative values and O. vulgaris slightly positive values, both values were close to zero, suggesting demographic stability in both species. The unimodal mismatch distributions observed for \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e matched the expected distributions under a sudden expansion model, supporting the neutrality test results. Conversely, \u003cem\u003eS. cabrilla\u003c/em\u003e and \u003cem\u003eO. vulgaris\u003c/em\u003e exhibited bimodal mismatch distributions (Fig. 5), strongly deviating from the expansion model.\u003c/p\u003e\n\u003cp\u003eFrom the coalescent simulation tests of bottleneck scenario, the observed nucleotide diversity at the non-protected SCI and ADJ zones for \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e was lower than the expected 95% upper CI limit of nucleotide diversity replicates generated from the simulation (Table 3). This suggests that the proposed model of population bottlenecks caused by fishing in the SCI and ADJ could explain the observed nucleotide diversity estimates. In contrast, the observed nucleotide diversity values for \u003cem\u003eS. cabrilla\u0026nbsp;\u003c/em\u003eand \u003cem\u003eO. vulgaris\u003c/em\u003e from the SCI and ADJ zones, although lower than in FPZ, did not fall within the 95% upper CI limit of nucleotide diversity estimates expected by the bottleneck model, suggesting no significant effect of the current fishing activities on the genetic diversity levels of these two species.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAccording to the hypothesis of population expansion after banning trawling activities in the protected zone (FPZ), the observed nucleotide diversity values were significantly higher than the expected values from a population expansion derived from the coalescent simulations for all species (Table 3). This could suggest that protection may have allowed genetic diversity recovery to levels higher than expected by our tested model. It could be because the nucleotide diversity in FPZ before protection was higher than the current diversity observed in the other two zones, or because growing rate is higher than in the model.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn the present study, we evaluated how a Fisheries Protection Zone in the Menorca Channel influences genetic diversity by analysing the mitochondrial COI gene in four exploited marine species, three teleost fishes (\u003cem\u003eMullus surmuletus\u003c/em\u003e, \u003cem\u003eSerranus cabrilla,\u003c/em\u003e and \u003cem\u003eScorpaena notata\u003c/em\u003e) and one cephalopod (\u003cem\u003eOctopus vulgaris\u003c/em\u003e). Overall, individuals collected from the FPZ displayed the highest nucleotide diversity values compared to those from non-protected zones, the closest SCI and the farthest ADJ. These results suggest a better conservation status for the studied species within the protected area.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRecent studies have reported positive effects from the establishment of the FPZ in the Menorca Channel, demonstrating that the 2016 trawling ban has helped protect 80% of rhodolith beds and 95% of coralligenous bottoms from trawling. This protection has led to increased biomass of rhodolith beds and \u003cem\u003eLaminaria rodriguezii\u003c/em\u003e forests, thereby contributing to the recovery of epibenthic communities (Farriols et al. 2021; 2025). The distribution of these habitats has increased by 6% and 54%, respectively, in recent years, highlighting the FPZ as an effective conservation measure for benthic habitats, communities, and species in the Menorca Channel (Farriols et al. 2025). The recovery of these significant marine habitats is enhancing marine biodiversity in the area due to their structural and functional complexity (Barberá et al. 2012; Joher et al. 2012, 2015), which benefits both exploited and unexploited species.\u003c/p\u003e\n\u003cp\u003eIn addition to the implementation of the FPZ, some areas of the Menorca Channel had already undergone previous changes. On the one hand, submarine cables were installed in the 1970s, in a narrow area within what is now the FPZ, connecting Mallorca and Menorca (Cabrito et al. 2024), which resulted in trawling being excluded from this area for decades, potentially providing long-term benefits to marine communities. On the other hand, a general reduction in fishing effort has been recorded across the Balearic Islands in recent decades (Quetglas et al. 2017).\u0026nbsp;Therefore, the recovery trend observed in this study may have originated from these prior changes and was further enhanced by the implementation of the protected area.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSpecimens of the four studied species collected from non-protected areas in the Menorca Channel showed high nucleotide diversity values, although lower compared to those from the protected area. This is to be expected given the geographical proximity to the FPZ, suggesting that adjacent areas may benefit from population growth in the protected zone. It is therefore a clear example of how a marine reserve can play a crucial role in sustaining the productivity of fishing zones and contributing to the consequent gene flow between protected and near non-protected areas, which is essential for preserving and restoring ecological processes and for ensuring genetic diversity, which supports species’ ability to adapt to global environmental changes (Gandra et al. 2020). In this sense, the lack of genetic structuring observed in the haplotype networks (Fig. 4) and the low and non-significant\u0026nbsp;Φ\u003csub\u003est\u0026nbsp;\u003c/sub\u003evalues\u0026nbsp;(Table S4) suggest high levels of gene flow between the FPZ and nearby zones. In the Mediterranean, similar patterns have been observed in fish species such as \u003cem\u003eDiplodus vulgaris\u003c/em\u003e, \u003cem\u003eMullus surmuletus\u003c/em\u003e and \u003cem\u003eM. barbatus\u003c/em\u003e, where high genetic diversity was found across both protected and adjacent fished areas (Félix-Hackradt et al. 2013; Sahyoun et al. 2016).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHowever, for the three teleost fish species analysed in this study, the lowest nucleotide diversity values were unexpectedly observed in the SCI, the non-protected zone closest to the protected FPZ. As this zone presents similar habitat characteristics to those of protected area (Farriols et al. 2025), the observed reduction in genetic diversity could be attributable to increased fishing pressure rather than habitat differences. Supporting this, Vessel Monitoring System (VMS) data indicate that, following the establishment of the FPZ, trawling activities in the Menorca Channel shifted mainly toward the southern area of the SCI (Fig. 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWhile all target species in this study exhibited higher genetic diversity values within the protected area, interspecific differences in nucleotide diversity levels were observed. Specifically, \u003cem\u003eS. cabrilla\u003c/em\u003e showed the highest nucleotide diversity, followed by\u003cem\u003e\u0026nbsp;O. vulgaris\u003c/em\u003e, whereas \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e presented the lowest values. These differences could be related to differences in the life-history traits of the species studied. A key distinction is \u003cem\u003eS. cabrilla\u003c/em\u003e’s reproductive strategy asa simultaneous hermaphrodite that does not self-fertilize, which reduces homozygosity and inbreeding risk, thereby enhancing genetic diversity by promoting spawning events and offspring heterozygosity (Avise and Mank 2009; Coscia et al. 2016). Since each individual in the population contributes mitochondrial DNA to the next generation—unlike in non-hermaphroditic species, where only females do—the resulting nucleotide diversity is not directly comparable to that of the other species. Regarding O. vulgaris, this species exhibits a fast growth rate, a short semelparous life cycle that allows rapid generational turnover,\u0026nbsp;and high fecundity (Hanlon and Messenger 1996), traits that confer relative resilience to fishing pressure and environmental perturbations compared with many teleost species \u003cem\u003e(Faure et al. 2000)\u003c/em\u003e. Lastly, M. surmuletus and S. notata possess typical life-history traits of marine fish: they are\u0026nbsp;oviparous species with\u0026nbsp;external fertilization, their eggs are embedded in a gelatinous matrix, and they have relatively short generation times—3.8 and 2.9 years, respectively (Table 4).\u003c/p\u003e\n\u003cp\u003eIn order to compare the nucleotide diversity estimates of the studied species in the Menorca Channel with those reported for other species across the Balearic Islands, only two studies were found (Petit-Marty et al. 2022; Riera et al. 2025), both focusing on teleost species (Fig. 6).\u0026nbsp;Compared with the estimate reported by Petit-Marty et al. (2022) (π = 0.0047),\u0026nbsp;\u003cem\u003eM. surmuletus\u003c/em\u003e exhibited lower nucleotide diversity in this study (π = 0.0026), whereas\u0026nbsp;\u003cem\u003eS. notata\u003c/em\u003e presented slightly greater diversity\u0026nbsp;(π = 0.0025) than the estimate for the entire population across the Balearic Archipelago (π = 0.0020; Riera et al. 2025). These intraspecific differences may reflect variations in sampling effort and geographic coverage across the Balearic Islands. Additionally, \u003cem\u003eM. surmuletus\u003c/em\u003e displayed lower nucleotide estimates than its sister species, \u003cem\u003eM. barbatus\u003c/em\u003e (π = 0.0035) (Fig. 6).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBoth \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e from the Menorca Channel exhibited nucleotide diversity values below the lower limit of the 95% confidence interval for Mediterranean fish species as calculated by Petit-Marty et al. (2022) (mean 95% CI = 0.0034–0.0047) (Fig. 6). In this context, both species exhibited genetic diversity levels slightly higher than two overexploited species, \u003cem\u003eM. merluccius\u003c/em\u003e (π = 0.0018) and \u003cem\u003eLophius budegassa\u003c/em\u003e (π = 0.0016)\u0026nbsp;(Fig. 6), both characterized by large body size and long generation times (~10 years) (Petit-Marty et al. 2022). Although the comparison should be interpreted with caution, as the mutation rate may vary among genera, these results suggest that both species could be affected by the impact of fishing. In the case of \u003cem\u003eS. notata\u003c/em\u003e, this aligns with the findings of Riera et al. (2025), who reported signs of fishing pressure on this species in the Balearic Islands, despite it being a bycatch species.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFinally, \u003cem\u003eS. cabrilla\u003c/em\u003e exhibited the highest values even exceeding the upper limit of the 95% confidence interval for Mediterranean fish species (Petit-Marty et al. 2022) (Fig. 6), as expected given its reproduction strategy described previously.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe genetic diversity values estimated in the four target species also varied when compared with those of the same species from other areas across the Mediterranean Sea and Northeast Atlantic. \u003cem\u003eM. surmuletus\u003c/em\u003e from the Menorca Channel (π = 0.0026) exhibited similar values to populations in Egypt (π = 0.0030) (Soliman et al. 2024). \u003cem\u003eS. cabrilla\u003c/em\u003e showed nucleotide diversity values\u0026nbsp;consistent with those reported in the Eastern Mediterranean (π = 0.0118), although populations from Turkey exhibited lower genetic diversity (π = 0.0035) (Bos et al. 2020).\u0026nbsp;Lastly, \u003cem\u003eO. vulgaris\u003c/em\u003e exhibited nucleotide diversity levels (π = 0.0045) consistent with estimates from Central Mediterranean (Sardinia, π = 0.0046) (Melis et al. 2018) and Northeast Atlantic populations (Portugal, π = 0.0040; and Canary Islands, π = 0.0059) (Pérez et al. 2023).\u003c/p\u003e\n\u003cp\u003eRegarding demographic history and coalescent simulations analyses, the target species displayed distinct patterns. Populations of \u003cem\u003eM. surmuletus\u003c/em\u003e and \u003cem\u003eS. notata\u003c/em\u003e in the Menorca Channel showed signs of recent expansion, as indicated by significant neutrality statistics and “star” or “complex”-star-shaped haplotype networks (Table 2, Fig. 4). Similar results have been observed in \u003cem\u003eS. notata\u0026nbsp;\u003c/em\u003epopulation across the Balearic Islands (Riera et al. 2025). In contrast, \u003cem\u003eS. cabrilla\u0026nbsp;\u003c/em\u003eand \u003cem\u003eO. vulgaris\u003c/em\u003e presented non-significant neutrality statistics and bimodal mismatch distributions, suggesting a stable population size. Bimodal distributions may also indicate complex population structures, such as the presence of distinct lineages (Jenkins et al. 2018), although this was not clearly evident in the haplotype networks. For octopus, two different mitochondrial haplotype lineages have been observed along the East Atlantic coast and locally along Sardinia’s coast (Melis et al. 2018; Quinteiro et al. 2020), as well as a complex genetic pattern across the Mediterranean (Fadhlaoui-Zid et al. 2021). Further analyses should be realised to explore the genetic structure and demographic dynamics of octopus and comber species in the Balearic Islands.\u003c/p\u003e\n\u003cp\u003eCoalescent simulations help assess whether observed results align with expected demographic events such as expansions or bottlenecks in the different zones studied within the Menorca Channel. Coalescent simulations indicated that FPZ populations grew more than expected by a population expansion if the levels of nucleotide diversity in the FPZ populations before protection were equal to those observed in the non-protected zones. The contrary was also true, bottleneck simulations showed that the observed values of nucleotide diversity in the fished zones were significantly lower than expected by a reduction of population starting with the levels observed in the FPZ population. Differences in genetic diversity between ADJ and SCI may stem from historically low diversity levels in the Menorca Channel prior to FPZ implementation. Before protection, high gene flow combined with fishing pressure likely homogenized genetic diversity across zones. However, the establishment of the FPZ enabled population growth within protected areas. In contrast, individuals outside the FPZ faced intense fishing pressure, leading to population decline. The impact was stronger in the closest SCI zone where trawling activities were concentrated (Fig. 2) likely leading to greater genetic erosion than in the farthest ADJ zone, where fishing efforts remained relatively stable following the establishment of the trawling ban. In this scenario, the fishing-driven decline in SCI may have exceeded the demographic benefits of gene flow from the FPZ, whereas the ADJ—although also affected—experienced milder losses.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn contrast, for \u003cem\u003eS. cabrilla\u003c/em\u003e and \u003cem\u003eO. vulgaris\u003c/em\u003e, coalescent simulations also suggested population expansion within the FPZ, but no decline in non-protected zones under the tested conditions. This pattern may result from a) a bottleneck less severe than modelled, b) a recent demographic decline not yet detectable through genetic signals, or c) no significant effect of current trawling activities on the genetic diversity of these species. Additionally, the relatively resilient life history traits of \u003cem\u003eS. cabrilla\u003c/em\u003e and \u003cem\u003eO. vulgaris\u003c/em\u003e—such as hermaphroditism in the former and rapid turnover in the latter—may have caused these species to recover more quickly from the effects of fishing. These characteristics likely contributed to the higher genetic diversity observed in the present study, making these two species\u0026nbsp;good indicators of conservation effectiveness in both protected and non-protected zones.\u003c/p\u003e\n\u003cp\u003eThis study provides genetic evidence of the positive impact of a Fisheries Protection Zone (FPZ) on overexploited marine species in the Menorca Channel, employing mitochondrial COI nucleotide diversity as a novel proxy for species conservation status. Although mitochondrial DNA has several limitations due to its characteristics—maternally inhered, haploidy and lack of recombination—it remains a valuable marker for genetic conservation studies. A positive correlation between mitochondrial and nuclear genetic diversity in fish species has been reported (Piganeau and Eyre-Walker 2009), suggesting that a reduction in mitochondrial COI diversity may reflect broader nuclear genomic erosion and, therefore, can provide useful insight into the conservation status of marine species populations.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOverall, our findings indicate a positive trend in the recovery of genetic diversity in exploited species, enhanced by the establishment of the Fisheries Protection Zone, which has contributed to the improved condition of marine habitats within the study area—improvements that are closely linked to the conservation status of marine species. Moreover, this study highlights the value of genetic diversity approaches as effective diagnostic tools to assess protected and non-protected zones, and to support ongoing monitoring of vulnerable and exploited species.\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study is part of the MARFISH project (PDR2020/69–ITS 2017-006), included in the \u003cem\u003eProjectes de Recerca Científica i Tecnològica\u003c/em\u003e of the \u003cem\u003eDirecció General de Política Universitària i Recerca\u003c/em\u003e, which are funded by the Comunitat Autònoma de les Illes Balears (CAIB) through the Direcció Conselleria de Fons Europeus, Universitat i Cultura, with resources from the Tourist Stay Tax Law. The CAIB also funded the 2022 grants for the training of research personnel (N. Pasini, research grant nº FPI_026_2022). The MEDITS surveys are cofunded by the European Union through the European Maritime Fisheries and Aquaculture Fund (EMFAF) within the National Program of collection, management, and use of data in the fisheries sector and support for scientific advice regarding the Common Fisheries Policy. The CANAL surveys were part of the SosMed project (Improvement in scientific and technical knowledge for the sustainability of demersal fisheries in the western Mediterranean), funded by the European Union—Next Generation (Recovery, Transformation and Resilience Plan), with an agreement between the Ministry of Agriculture, Fisheries and Food and the Spanish National Research Council, through the Spanish Institute of Oceanography.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConceptualization, F.O., A.P. and S.R.-A.; methodology, N.P.; data analysis, N.P., S.R.-A., M.B., J.F.F, M.T.F. and N.P.-M.; writing—original draft preparation, N.P.; writing—review and editing, M.B., J.F.F., M.T.F., N.P.-M., F.O., S.R.-A. and A.P.; funding acquisition, F.O. and A.P. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data supporting the findings of this study are available within the paper and its Supplementary Information. COI mitochondrial sequences were deposited into the GenBank (NCBI) database under accession numbers: PQ339824-PQ339921 (\u003cem\u003eMullus surmuletus\u003c/em\u003e); PQ339926-PQ340023 (\u003cem\u003eSerranus cabrilla\u003c/em\u003e); PQ363178-PQ363270 (\u003cem\u003eScorpaena notata\u003c/em\u003e); and PQ340025-PQ340124 (\u003cem\u003eOctopus vulgaris\u003c/em\u003e).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAllcock L, Headlam J, Allen G (2018) Octopus vulgaris. 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Estuar Coast Shelf Sci 85:537\u0026ndash;546. https://doi.org/10.1016/j.ecss.2009.09.020 \u003c/li\u003e\n\u003cli\u003eOrdines F, Bauz\u0026aacute; M, Sbert M, et al (2015) Red algal beds increase the condition of nekto-benthic fish. J Sea Res 95:115\u0026ndash;123. https://doi.org/10.1016/j.seares.2014.08.002 \u003c/li\u003e\n\u003cli\u003eP\u0026eacute;rez T, Romero-Bascones A, Pirhadi N, et al (2023) Insights on the Evolutionary History and Genetic Patterns of Octopus vulgaris Cuvier, 1797 in the Northeastern Atlantic Using Mitochondrial DNA. Animals 13:2708. https://doi.org/10.3390/ani13172708 \u003c/li\u003e\n\u003cli\u003ePetit-Marty N, V\u0026aacute;zquez-Luis M, Hendriks IE (2020) Use of the nucleotide diversity in COI mitochondrial gene as an early diagnostic of conservation status of animal species. Conserv Lett 14:1\u0026ndash;7. https://doi.org/10.1111/conl.12756 \u003c/li\u003e\n\u003cli\u003ePetit-Marty N, Liu M, Tan IZ, et al (2022) Declining Population Sizes and Loss of Genetic Diversity in Commercial Fishes: A Simple Method for a First Diagnostic. Front Mar Sci 9:1\u0026ndash;11. https://doi.org/10.3389/fmars.2022.872537 \u003c/li\u003e\n\u003cli\u003ePiganeau G, Eyre-Walker A (2009) Evidence for Variation in the Effective Population Size of Animal Mitochondrial DNA. PLoS One 4:e4396. https://doi.org/10.1371/journal.pone.0004396 \u003c/li\u003e\n\u003cli\u003ePinsky ML, Palumbi SR (2014) Meta-analysis reveals lower genetic diversity in overfished populations. Mol Ecol 23:29\u0026ndash;39. https://doi.org/10.1111/mec.12509 \u003c/li\u003e\n\u003cli\u003eQuetglas A, Alemany F, Carbonell A, et al (1998) Biology and fishery of Octopus vulgaris Cuvier, 1797, caught by trawlers in Mallorca (Balearic Sea, Western Mediterranean). 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Journal of Marine Science and Engineering 11:1982. https://doi.org/10.3390/jmse11101982 \u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Genetic and haplotype diversity indices of COI marker in\u003cem\u003e\u0026nbsp;\u003c/em\u003efour species across the three studied zones: the Fishery Protection Zone (FPZ), where trawling is banned; the Site of Community Importance (SCI), the nearest open-to-fishing zone to the FPZ; and the ADJ zone, the farthest open-to-fishing area from the FPZ. Amplified COI fragment lengths: \u003cem\u003eMullus surmuletus\u003c/em\u003e = 600 bp, \u003cem\u003eSerranus cabrilla\u003c/em\u003e = 612 bp, \u003cem\u003eSerranus notata\u003c/em\u003e = 642 bp, \u003cem\u003eOctopus vulgaris\u003c/em\u003e = 657 bp. Symbols are defined in the footnote\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"633\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 56px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZONE\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eh\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHd \u0026plusmn; SD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eKt\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026pi; \u0026plusmn; SD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" style=\"width: 56px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eMullus surmuletus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eFPZ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.6987 \u0026plusmn; 0.0740\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1.6894\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0028 \u0026plusmn; 0.0004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eSCI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.6292 \u0026plusmn; 0.0630\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1.2674\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0021 \u0026plusmn; 0.0002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eADJ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.7505 \u0026plusmn; 0.0500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1.6430\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0027 \u0026plusmn; 0.0003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e98\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e20\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.6933 \u0026plusmn; 0.0360\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.5464\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0026 \u0026plusmn; 0.0002\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" style=\"width: 56px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSerranus cabrilla\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eFPZ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.8712 \u0026plusmn; 0.0290\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e5.8371\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0095 \u0026plusmn; 0.0006\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eSCI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.7825 \u0026plusmn; 0.0630\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e5.3957\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0088 \u0026plusmn; 0.0009\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eADJ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.8645 \u0026plusmn; 0.0390\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e5.5828\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0091 \u0026plusmn; 0.0006\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e98\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e28\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e21\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.8502 \u0026plusmn; 0.0200\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e5.5620\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0091 \u0026plusmn; 0.0004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" style=\"width: 56px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eScorpaena notata\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eFPZ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.7954 \u0026plusmn; 0.0680\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e2.2643\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0035 \u0026plusmn; 0.0007\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eSCI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.6237 \u0026plusmn; 0.0990\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e0.8086\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0013 \u0026plusmn; 0.0003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eADJ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.8528 \u0026plusmn; 0.0570\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e1.5806\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0025 \u0026plusmn; 0.0004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e93\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e35\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e32\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.7632 \u0026plusmn; 0.0460\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.5680\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0024\u0026plusmn; 0.0003\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" valign=\"top\" style=\"width: 56px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eOctopus vulgaris\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eFPZ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.7770 \u0026plusmn; 0.0380\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3.5012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0053 \u0026plusmn; 0.0004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eSCI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.7683 \u0026plusmn; 0.0450\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3.4029\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0052 \u0026plusmn; 0.0003\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eADJ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e0.7492 \u0026plusmn; 0.0460\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e3.3397\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e0.0051 \u0026plusmn; 0.0005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e14\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 57px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.7713 \u0026plusmn; 0.0180\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.3851\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0052 \u0026plusmn; 0.0002\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003ea\u003c/sup\u003eN = number of individuals; S = number of segregating sites; h = number of haplotypes; Hd = haplotype diversity; Kt = mean nucleotide difference; \u0026pi; = nucleotide diversity; SD = standard deviation\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e Tajima\u0026rsquo;s D, Fu\u0026rsquo;s FS, Ramos-Onsins and Rozas\u0026apos;s R\u003csub\u003e2\u003c/sub\u003e indices, as well as mismatch distribution metrics (including the sum of squared deviations from the sudden expansion model, SSD, and the Harpending raggedness index, rg), are indicated for each species. The corresponding p-values are reported in brackets. Values in bold were significant at the 95% confidence level\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"680\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eR\u003csub\u003e2\u003c/sub\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSSD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003erg\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eMullus surmuletus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-1.7452 (\u003cstrong\u003e0.0145\u003c/strong\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-10.6423 (\u003cstrong\u003e0.0002\u003c/strong\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e0.0371 (\u003cstrong\u003e0.0391\u003c/strong\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0261 (0.2509)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0883 (0.1621)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSerranus cabrilla\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-0.0332 (0.6000)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-1.9033 (0.3163)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e0.0958 (0.5727)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0307 (0.2609)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0333 (0.4172)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eScorpaena notata\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-2.4020 (\u003cstrong\u003e0.0000\u003c/strong\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e-28.0227 (\u003cstrong\u003e0.0000\u003c/strong\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e0.0210 (\u003cstrong\u003e0.0000\u003c/strong\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0134 (0.2220)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0758 (0.2260)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eOctopus vulgaris\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e0.6886 (0.8018)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003e0.8488 (0.6770)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e0.1173 (0.7849)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.0786 (0.0537)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.1050 (0.1579)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e Observed (\u0026pi;\u003csub\u003eob\u003c/sub\u003e) and expected (\u0026pi;\u003csub\u003eex\u003c/sub\u003e)\u003csub\u003e\u0026nbsp;\u003c/sub\u003enucleotide diversity values are shown for each species in the three zones: the Fishery Protection Zone (FPZ), where trawling is banned; the Site of Community Importance (SCI), the nearest open-to-fishing zone to the FPZ; and the ADJ zone, the farthest open-to-fishing area from the FPZ. Observed values were calculated using DnaSP v6. Expected values were modelled through coalescent simulations assuming equal genetic diversity across zones prior to the FPZ\u0026rsquo;s establishment. Each simulation used \u003cem\u003e\u0026theta;\u0026nbsp;\u003c/em\u003e(\u003cem\u003e\u0026theta;\u003c/em\u003e = 4Neu) as starting parameter, estimated by the observed Watterson\u0026rsquo;s estimator (\u003cem\u003e\u0026theta;\u003csub\u003eW\u003c/sub\u003e\u003c/em\u003e) in FPZ samples (for bottleneck scenarios) and by the observed average \u003cem\u003e\u0026theta;\u003csub\u003eW\u003c/sub\u003e\u003c/em\u003e in the non-protected zones SCI and ADJ specimens (for expansion scenarios), with 1,000 random samplings of \u003cem\u003eN\u003c/em\u003e individuals per species (\u003cem\u003eN\u003c/em\u003e = observed sample size). Bottleneck intensity was calculated as the ratio of nucleotide diversity in individuals from SCI or ADJ (\u0026pi;\u003csub\u003eS\u003c/sub\u003e and \u0026pi;\u003csub\u003eA\u003c/sub\u003e, respectively) to those from FPZs (\u0026pi;\u003csub\u003eF\u003c/sub\u003e); expansion intensity was calculated as the ratio of \u0026pi;\u003csub\u003eF\u003c/sub\u003e to the average nucleotide diversity in non-protected areas (\u0026pi;\u003csub\u003eS-A\u003c/sub\u003e). The 95% upper CI limit of the 1,000 nucleotide diversity replicates obtained from the msms program are indicated\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\" style=\"margin-left: calc(0%); width: 100%;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"9\" valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eBottleneck simulations\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"bottom\" style=\"width: 6.5682%;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46.6338%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eMullus surmuletus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSerranus cabrilla\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eBottleneck intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eBottleneck intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.74822\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00211\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00332\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e0.00345\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.95597\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00882\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e0.00725\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00750\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eADJ\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.97163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00274\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00427\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e0.00443\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.92453\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00912\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e0.00766\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00794\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46.6338%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eScorpaena notata\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eOctopus vulgaris\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eBottleneck intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eBottleneck intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSCI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.69688\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00270\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e0.00282\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.95310\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00518\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e0.00348\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00361\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eADJ\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.35694\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00246\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00482\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e0.00499\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.97186\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00508\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e0.00339\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00353\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"9\" valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003ePopulation expansion simulations\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46.6338%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eMullus surmuletus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSerranus cabrilla\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eExpansion intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eExpansion intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFPZ\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e1.16289\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00282\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00232\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e0.00240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e1.06354\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00954\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e0.00535\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00550\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46.6338%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eScorpaena notata\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eOctopus vulgaris\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eExpansion intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003eExpansion intensity\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eob\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e\u0026pi;\u003csub\u003eex\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cem\u003e95% upper CI limit\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 6.5682%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFPZ\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e1.89785\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00353\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00303\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.8374%;\"\u003e\n \u003cp\u003e0.00310\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e1.03899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00533\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e0.00243\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00250\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4\u003c/strong\u003e Characteristics of the studied species. \u003cem\u003eMax. size\u003c/em\u003e and \u003cem\u003egen. times\u003c/em\u003e define the maximum recorded size in centimetres and the mean generation time in years, respectively. Information on commercial importance, fishing gear, IUCN status, and movement/habitats was obtained from IUCN reports for the evaluated species and their populations in the Mediterranean. Data on maximum size and generation times were retrieved from FishBase. N/A= not available\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"962\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCommercial importance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFishing gear\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIUCN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 132px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePopulation trends\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eThreats\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 178px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMovements/Habitats\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMax. size (cm)/gen. times (year)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eMullus surmuletus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eHigh\u003c/p\u003e\n \u003cp\u003e(Carpenter et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003eBottoms trawls, trammel nets\u003c/p\u003e\n \u003cp\u003e(Carpenter et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eLC\u003c/p\u003e\n \u003cp\u003e(Carpenter et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 132px;\"\u003e\n \u003cp\u003eDecreasing\u003c/p\u003e\n \u003cp\u003e(Quetglas et al. 2016)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 142px;\"\u003e\n \u003cp\u003eOverexploitation\u003c/p\u003e\n \u003cp\u003e(FAO 2024)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 178px;\"\u003e\n \u003cp\u003eOceanodromous/\u003c/p\u003e\n \u003cp\u003eMarine oceanic - Epipelagic (0-200 m)\u003c/p\u003e\n \u003cp\u003e(Carpenter et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e40.0 cm/3.8 years\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSerranus cabrilla\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMinor\u003c/p\u003e\n \u003cp\u003e(Smith-Vaniz and Iwamoto 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003eRecreational fishing; bycatch in gillnet, bottom longline and trawls\u003c/p\u003e\n \u003cp\u003e(Smith-Vaniz and Iwamoto 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eLC\u003c/p\u003e\n \u003cp\u003e(Smith-Vaniz and Iwamoto 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 132px;\"\u003e\n \u003cp\u003eStable\u003c/p\u003e\n \u003cp\u003e(Smith-Vaniz and Iwamoto 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 142px;\"\u003e\n \u003cp\u003eOverexploitation\u003c/p\u003e\n \u003cp\u003e(Quetglas et al. 2016)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 178px;\"\u003e\n \u003cp\u003eLow migrant/\u003c/p\u003e\n \u003cp\u003eMarine neritic - Subtidal rock, sandy-mud\u003c/p\u003e\n \u003cp\u003e(Smith-Vaniz and Iwamoto 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e40.0 cm/5.7 years\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eScorpaena notata\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eMinor\u003c/p\u003e\n \u003cp\u003e(Nunoo et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003eBycatch in trammel net and bottom trawls\u003c/p\u003e\n \u003cp\u003e(Nunoo et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eLC\u003c/p\u003e\n \u003cp\u003e(Nunoo et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 132px;\"\u003e\n \u003cp\u003eN/A\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 142px;\"\u003e\n \u003cp\u003eN/A\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 178px;\"\u003e\n \u003cp\u003eLow migrant/\u003c/p\u003e\n \u003cp\u003eMarine neritic \u0026ndash; deep benthic\u003c/p\u003e\n \u003cp\u003e(Nunoo et al. 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e26.0 cm/2.9 years\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eOctopus vulgaris\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003eHigh\u003c/p\u003e\n \u003cp\u003e(Allcock et al. 2018)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003eTraps and spots, trammel nets, bottom trawls\u003c/p\u003e\n \u003cp\u003e(Allcock et al. 2018)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eLC\u003c/p\u003e\n \u003cp\u003e(Allcock et al. 2018)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 132px;\"\u003e\n \u003cp\u003eN/A\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 142px;\"\u003e\n \u003cp\u003eOverexploitation\u003c/p\u003e\n \u003cp\u003e(FAO 2024)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 178px;\"\u003e\n \u003cp\u003eMigrant/\u003c/p\u003e\n \u003cp\u003eMarine neritic - Subtidal rock, sandy-mud, macroalgal/kelp\u003c/p\u003e\n \u003cp\u003e(Allcock et al. 2018)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 123px;\"\u003e\n \u003cp\u003e180 cm/1.2 years\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"biodiversity-and-conservation","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bioc","sideBox":"Learn more about [Biodiversity and Conservation](https://www.springer.com/journal/10531)","snPcode":"10531","submissionUrl":"https://submission.nature.com/new-submission/10531/3","title":"Biodiversity and Conservation","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Nucleotide diversity, COI, Coalescent simulations, Fisheries resources, Protected areas, Mediterranean Sea","lastPublishedDoi":"10.21203/rs.3.rs-7271616/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7271616/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOverexploitation can drive evolutionary changes and erode genetic diversity, reducing species\u0026rsquo; adaptive capacity to environmental and anthropogenic pressures. Spatial marine conservation measures, such as Marine Protected Areas and Fisheries Protection Zones (FPZs), aim to mitigate these impacts by preserving biodiversity and promoting sustainable fisheries. Recently, nucleotide diversity of the mitochondrial Cytochrome C Oxidase subunit I (COI) marker has emerged as a promising proxy for assessing species conservation status. To evaluate the effectiveness of an FPZ established in 2016 in the Menorca Channel, COI genetic diversity was assessed in four exploited marine species across three areas: the FPZ and two nearby non-protected zones. All species exhibited consistently higher genetic diversity within the FPZ, despite evidence of high gene flow among areas. Coalescent simulations were used to model expected genetic diversity under neutral scenarios of bottlenecks and expansions, with magnitudes estimated from differences in nucleotide diversities observed between fished and non-fished zones. Simulations supported a scenario of population expansion in the FPZ, contrasting with signs of genetic erosion in fished areas. These patterns align with Vessel Monitoring System (VMS) data, which show a post-protection-establishment shift in fishing effort toward non-protected zones, potentially contributing to population declines outside the FPZ. This study provides genetic evidence of the positive effects of fishing restrictions on fishery resources in the Menorca Channel, supporting the FPZ\u0026rsquo;s role in preserving genetic diversity and promoting population recovery. Furthermore, it highlights COI nucleotide diversity as a simple, cost-effective tool for monitoring marine species\u0026rsquo; conservation status and guiding resource management strategies.\u003c/p\u003e","manuscriptTitle":"Genetic diversity in mitochondrial DNA reveals the effect of a Fisheries Protection Zone on exploited marine species in the Menorca Channel (Western Mediterranean)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-13 17:01:33","doi":"10.21203/rs.3.rs-7271616/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-06T14:56:36+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-12T07:57:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-25T12:55:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"93226325355041545547319776528261325543","date":"2025-11-10T08:00:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"117040331901566187225991942704088741887","date":"2025-11-04T14:56:47+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"88694160820685715675192302418477047549","date":"2025-11-04T08:19:04+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-04T07:08:50+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-24T10:52:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-05T15:12:40+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biodiversity and Conservation","date":"2025-08-01T13:11:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biodiversity-and-conservation","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bioc","sideBox":"Learn more about [Biodiversity and Conservation](https://www.springer.com/journal/10531)","snPcode":"10531","submissionUrl":"https://submission.nature.com/new-submission/10531/3","title":"Biodiversity and Conservation","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"be842a7e-9884-4339-aa81-15e6df434b8f","owner":[],"postedDate":"November 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-31T14:53:13+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-13 17:01:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7271616","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7271616","identity":"rs-7271616","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}