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. 2021 Sep 21;16(9):e0234534. doi: 10.1371/journal.pone.0234534

Total mercury concentrations in invasive lionfish (Pterois volitans/miles) from the Atlantic coast of Florida

Eric G Johnson 1,*,#, Angelina Dichiera 1,¤a,#, Danielle Goldberg 1,¤b, MaryKate Swenarton 1,¤c, James Gelsleichter 1
Editor: Myra E Finkelstein2
PMCID: PMC8454969  PMID: 34547024

Abstract

Invasive lionfish (Pterois volitans/miles) pose a serious threat to marine ecosystems throughout the western Atlantic Ocean and Caribbean Sea. The development of a fishery for lionfish has been proposed as a strategy for controlling populations; however, there is concern about consumption of this species by humans due to its high trophic position and potential for bioaccumulation of mercury. We analyzed total mercury (THg) in tissues of lionfish from two locations on the east coast of Florida. THg in lionfish increased with size and differed by location and sex. THg was highest in muscle tissue and was strongly positively correlated among tissues. THg in lionfish was lower than other commonly consumed marine fishes, and falls into Florida’s least restrictive advisory level. Consumption of lionfish poses a low risk and concerns over mercury bioaccumulation should not present a significant barrier to lionfish harvest.

Introduction

Biological invasions are a significant and growing threat to global ecosystems causing both ecological harm and staggering economic costs [1, 2]. A marine invader of particular concern in the western Atlantic Ocean is the Indo-Pacific lionfish (a species continuum of two morphologically indistinct species, Pterois miles and P. volitans or potentially a hybrid of the two; [3]). Following introduction into the coastal waters of south Florida beginning over three decades ago [46], a combination of favorable life history traits has enabled this species to become among the most conspicuous and abundant residents in coastal ecosystems throughout the region [79].

Lionfish have long venomous spines that deter predation by native predators [10] and are resistant to common parasites [11]; however a recently discovered outbreak of an ulcerative skin disease in lionfish in the Gulf of Mexico suggests some susceptibility to disease [12]. Lionfish also inhabit a wide range of habitats and environmental conditions [1315], and are capable of long distance dispersal during pelagic egg and larval phases [16, 17] facilitating range expansion and colonization. Lionfish consume a generalist diet and are voracious predators of an array of reef fishes [18, 19]. Lionfish have been shown to reduce native fish recruitment [20] and overall native species biomass [21] in some studies, but not in others [22]. Lionfish directly impact recreationally and commercially important fishes by preying on them as juveniles [19, 23, 24] and impact adults of these same species indirectly through competition for food resources [24, 25].

To mitigate the impacts of lionfish on native ecosystems numerous removal strategies have been proposed [26]. Among these, harvest of lionfish by recreational and commercial spearfishers has shown promise as one method to help control populations of this invasive species [18, 2735]. Such efforts are being actively promoted by management agencies throughout the western Atlantic Ocean and Caribbean Sea, but require careful implementation to be most effective [33]. However, although lionfish have a white flesh with flavor and texture similar to highly-valued species such as snapper and grouper; efforts to develop a viable fishery for lionfish have been hampered by concerns that the concentration of biologically derived toxins, pollutants and heavy metals in this predator may be high enough to present an exposure risk for humans [36]. In Florida, mercury contamination is of particular concern, with high levels of this toxin found in many species of fishes [3739].

Mercury is a naturally occurring toxic metal that is known to bioaccumulate in the tissue of fishes. As mercury is primarily obtained from the diet, mercury can be magnified in the tissues of aquatic and marine predators, like lionfish, that feed at higher trophic levels [9, 25, 40]. The accumulation of mercury can not only directly impact fish health [41] and adversely affect many aspects of reproduction [42]. Mercury also poses a serious exposure risk for humans who consume fish, particularly for young or pregnant individuals [4345]. Further, most of the mercury present in fish muscle is present as organic methylmercury (MeHg) [4648], the most highly toxic and bioactive form [40]. MeHg exposure in humans is almost exclusively from the consumption of fish [49, 50]. To limit human exposure to mercury, dietary guidelines for fish consumption have been established (U.S. EPA, Florida DOH) and because rates of mercury bioaccumulation vary as a function of biological, environmental, and temporal factors, recommended levels of consumption often differ across species, sizes and locations.

The goal of this study was to quantify total mercury (THg) in lionfish as a function of capture location, sex, size and tissue type; information that is critical for evaluating the potential risk to consumers of this species in a rapidly developing fishery. Previous studies have provided assessments for mercury risk in lionfish [5154]; however, none of these studies were conducted in our region of study. Moreover, only one of the aforementioned studies examined mercury in lionfish across the entire range of sizes currently being harvested [51]. This study builds on earlier work by (1) expanding the spatial coverage to include an assessment of mercury in lionfish from unstudied regions, (2) expanding the range of sizes examined (particularly large individuals which have the highest potential for mercury bioaccumulation), and (3) quantifying mercury levels in different lionfish tissues to assess potential impacts on lionfish health and reproduction.

Materials and methods

Ethics statement

All lionfish used in this study were handled in strict accordance with a UNF IACUC protocol (IACUC#13–004) and tissues of opportunity waivers approved by the University of North Florida. UNF IACUC defines tissues of opportunity as samples collected: (1) during the course of another project with an approved IACUC protocol from another institution; (2) during normal veterinary care by appropriately permitted facilities; or (3) from free-ranging animals by appropriately permitted facilities. Lionfish removals are encouraged by the State of Florida and sample collection locations did not require any specific permissions. No endangered or protected species were harmed during the course of this study.

Field collections

Lionfish were collected in coordination with four recreational fishing tournaments (also termed derbies): three in northeast Florida (NEF; August 2013, April 2014 and August 2014) and one in southeast Florida (SEF; August 2014; Fig 1). Study regions (Fig 1) are representative, but only approximate, locations of capture since actual coordinates were not provided by tournament fishers. During each derby, teams of recreational divers captured lionfish from local sites and then returned to tournament headquarters where fish were counted, measured and weighed. Lionfish were then separated by location of capture and placed on ice for transport to the laboratory. Lionfish were either dissected fresh or frozen whole and stored in freezers at -20°C until later processing and mercury analysis.

Fig 1. Map of study locations.

Fig 1

Location of lionfish collection sites from northeast (NEF) and southeast (SEF) Atlantic coasts of Florida. The final figure was modified from a base map created using the USGS National Map Viewer (public domain): http://viewer.nationalmap.gov/viewer/.

Sample processing

Lionfish were processed following standard protocols for mercury analysis to minimize the potential for cross-contamination among samples (U.S. EPA, 2000). Each fish was measured for standard length (SL, mm) and total length (TL, mm), weighed (g), and sexed (Table 1). Sex determination was not possible for many smaller immature individuals [29] and not available for a limited number of lionfish (n = 5) which sustained extensive damage during capture and field processing. To represent the portion of the fish most often consumed by humans, muscle was collected from the fillet of the left side of each fish, just above the lateral line [37]. For a subset of male (n = 26) and female fish (n = 31), we collected additional samples of liver, adipose and ovarian tissue for analysis. All samples were dried in a 60°C oven for 48 hours (a duration sufficient to achieve constant weight of samples) then homogenized using a mortar and pestle prior to analysis of total mercury concentrations (hereafter THg). The proportion of methylmercury (CH3Hg+) was not quantified; however, THg provides a reasonable approximation for finfish muscle in which greater than 95% of total mercury is methylated [47, 55]. Unfortunately, the relationship between methylmercury and THg in other tissues is more variable [5658] thus THg may not be a good proxy for methylmercury and the concentrations for other tissues in this study are best considered as maximum possible values.

Table 1. Morphometric and THg summary data by location and sex for lionfish in Florida.

Location   Morphometrics THg dw (μg g-1) THg ww (μg g-1
A. Northeast Florida n SL (mm) TL (mm) Mass (g) THg (dw) Min Max THg (ww) Min Max
Male 50 206 ± 45 281 ± 57 382 ± 231 0.27 ± 0.16 0.06 1.05 0.05 0.01 0.21
Female 31 167 ± 38 245 ± 38 227 ± 115 0.26 ± 0.16 0.12 0.81 0.05 0.02 0.16
Unknown 33 148 ± 67 246 ± 38 148 ± 176 0.21 ± 0.17 0.09 0.99 0.04 0.02 0.20
All 114 178 ± 56 250 ± 72 250 ± 72 0.25 ± 0.16 0.06 1.05 0.05 0.01 0.21
B. Southeast Florida
Male 38 178 ± 40 267 ± 62 285 ± 188 0.24 ± 0.26 0.11 1.30 0.05 0.02 0.26
Female 22 165 ± 27 247 ± 38 260 ± 148 0.32 ± 0.53 0.11 3.28 0.06 0.02 0.65
Unknown 1 65 110 12 0.17 0.17 0.17 0.03 0.03 0.03
All 61 167 ± 35 167 ± 35 167 ± 35 0.29 ± 0.44 0.11 3.28 0.06 0.02 0.65
Total 175 174 ± 50 250 ± 66 271 ± 202 0.26 ± 0.29 0.06 3.28 0.05 0.01 0.65

Mean (± sd) values for sample size (n), fish morphometrics: (Standard length (SL), Total length (TL) and Mass (g)), and raw total mercury concentration (μg g-1) on a dry weight and wet weight basis for lionfish collected in two regions along Florida’s Atlantic coast. THg dry weight was converted to wet weight based on a simple linear regression model (WW = 0.1972*DW); only mean, minimum and maximum values are shown. For each region, data are summarized for all individuals and by sex.

Mercury analyses

THg in lionfish tissues were measured using a Direct Mercury Analyzer (DMA-80; Milestone Inc., Shelton, CT, USA); a method recognized by the U.S. EPA (EPA Method 7473). The DMA-80 was calibrated weekly using serial dilutions from a liquid standard (1000 mg/L ± 5 mg/L, Certified Reference Material (CRM), ASSURANCE, SPEX CertiPrep, Inc.).

To facilitate comparisons with previous work and to evaluate mercury levels relative to national [44] and local [59] guidelines for consumption, we converted THg in muscle tissue from a dry weight to wet basis using a simple linear regression model fit to a subsample of fish for which sample weights were available both before and after drying. The slope of the line allowed for percent moisture to be calculated and THg dry weight to be converted to a wet weight basis for unknown samples.

Quality assurance and control (QA/QC) protocols were rigidly followed to ensure acceptable levels of accuracy and precision in the data. Weekly calibration of the DMA-80 with liquid mercury standard yielded standard curves with r2 values > 0.99. Method blanks returned THg concentrations well below the lowest recorded value from any fish sample in the study (<10% of the lowest value recorded in the study; EPA method 7473) confirming that samples were not substantially affected by contamination. Standard blanks using a solid standard (CRM, NIST Standard Ref 2709a) were all within expected values, 0.9 ± 0.2 μg g-1. The solid standard was chosen because no fish tissue standard was available at the time of analysis; however subsequent paired analyses of the solid standard and fish protein standard (DORM-4, National Research Council of Canada) using the same protocol demonstrated that both standards fell within their respective expected ranges. Duplicate (n = 66) and triplicate (n = 34) tissue samples from the same individual were run a minimum of every 10 samples over the course of the study and yielded coefficients of variation (CVs) averaging 2.17% indicating acceptable levels of precision.

Statistical analysis

All statistical analyses were run within SAS 9.4 (SAS Institute), SPSS Statistics 25 for Windows (IBM) or SigmaPlot (Systat Software) software packages. When necessary THg data were log-transformed (ln THg) prior to analyses to satisfy the assumptions of normality (Kolmogorov-Smirnov test) and homogeneity of variances (Levene’s test) required for parametric statistical tests. Temporal differences in THg in lionfish from NEF were assessed using a one-way analysis of covariance (ANCOVA) with collection date as a main factor and standard length (SL) as a covariate. SL was chosen as the best covariate for all analyses because body mass was not available for all fish and TL is more likely to be biased by potential damage to the caudal fin occurring naturally or during capture, storage and processing. Potential differences in THg between locations (NEF and SEF) and sexes (male and female) were tested using a two-way ANCOVA model with SL as a covariate. In cases where interactions were present between model main effects and covariates, we applied the Johnson-Neyman procedure as suggested by Wilcox [60].

Differences in THg between sexes and among tissues were assessed using a two-way repeated measures ANCOVA (RM ANCOVA) for three tissues (muscle, liver, adipose) excluding ovarian tissue. A one-way RM ANCOVA model was also run for females separately to include ovary tissue in the analysis. In all cases, we accounted for violations of the assumption of sphericity by adjusting model degrees of freedom [61]. Following significant results from ANCOVA, a post-hoc multiple comparison test (Dunn-Šidák, [62]). was applied to determine significant differences among tissues. The relationship among THg in various tissues was assessed using correlation (Pearson’s r) for females and males independently. To maintain experimentwise error rate (nine total comparisons), we applied the sequential Bonferroni correction when assessing significance [63].

Results

A total of 175 samples of lionfish muscle tissue were analyzed for THg. The samples included 88 males, 53 females and 34 individuals for which sex could not be determined. Overall, fish ranged widely in length (SL: 57 to 306 mm, x¯ = 175 mm, TL: 83 to 411mm, x¯ = 250 mm), weight (4.5 to over 1000g, x¯ = 271g) and THg (0.01 to 3.28 μg g-1 dw; x¯ = 0.26; Table 1).

To compare our results to earlier work and to recommended guidelines for consumption, we converted THg in muscle tissue from a dry weight to wet weight basis using a simple linear regression model. The linear relationship between THg wet weight and dry weight (WW = 0.19*DW) was highly significant (r2 = 0.99, p <0.0001); no intercept was included in the final linear model since it was not significantly different from 0 (t = 0.002, p = 0.998). The percent moisture in lionfish muscle averaged 80.4 ± 1.8% with high precision (CV = 0.02) and was independent of fish size, sex, and location. Converted THg on a wet weight basis is provided in Table 1.

Collection time had no significant effect on THg (F[2,57] = 1.05, p = 0.37), thus data for NEF were pooled across time for subsequent analyses. Our initial two-way ANCOVA model revealed several significant interactions (p <0.05) among location and SL (Table 2A). To aid with interpretation of the data and to remove potentially confounding interactions, we ran ANCOVA models for each location separately (Table 2B and 2C). In these reduced models, ANCOVA revealed no significant differences in THg by sex at either site (Table 2B and 2C, Fig 2), although females had generally higher THg for a given size. A one-way ANCOVA model with sexes pooled revealed a highly significant positive relationship between SL and THg and a significant effect of location (Table 2D, Fig 3). Additionally, a significant interaction indicated that the regression slopes for the two locations were not the same (Table 2B). Further analysis (Johnson-Neyman procedure; [60]) determined that individuals greater than 196 mm SL were significantly higher in THg in SEF than in NEF, while smaller individuals were not significantly different between locations (Fig 3).

Table 2. Analyses of covariance (ANCOVA) tables for ln total mercury (μg g-1) concentrations in lionfish muscle tissue.

A. Full model
Source df SS MS F p
Location 1 0.94 0.94 6.97 0.009
Sex 1 0.09 0.09 0.66 0.417
SL 1 21.64 27.69 149.22 0.000
Location × Sex 1 0.44 0.44 5.51 0.074
Location × SL 1 1.08 1.08 10.57 0.005
Sex × SL 1 0.97 0.97 5.83 0.058
Location × Sex × SL 1 0.44 0.44 5.41 0.073
Error 132 17.73 0.13
B. Northeast Florida df SS MS F p
Sex 1 0.09 0.09 0.89 0.349
SL 1 9.94 9.94 95.94 0.000
Sex × SL 1 0.00 0.00 0.01 0.922
Error 77 7.98 0.10
C. Southeast Florida df SS MS F p
Sex 1 0.36 0.36 2.01 0.162
SL 1 12.05 12.05 119.68 0.000
Sex × SL 1 0.71 0.71 3.94 0.052
Error 55 9.76 0.18
D. Sexes combined df SS MS F p
Location 1 2.31 2.31 12.72 0.000
SL 1 16.54 16.54 91.02 0.000
Location × SL 1 2.47 2.47 13.57 0.000
Error 171 22.01 0.17

Output from Analyses of covariance (ANCOVA) models of ln total mercury (μg g-1) concentrations in lionfish muscle tissue from A. Both locations, B. Northeast Florida (NEF), and C. Southeast Florida (SEF) D. Sexes combined.

Fig 2. Relationships between lionfish and THg concentration by sex.

Fig 2

Linear regression relationships between size (Standard length) and total mercury concentration for female (solid circles, solid line) and male (open circles, dotted line) lionfish in northeast Florida (A) and southeast Florida (B).

Fig 3. Relationships between lionfish and THg concentration by study location.

Fig 3

Linear regression relationships between size (Standard length) and total mercury concentration in lionfish from NEF (solid circles, solid line) and SEF (open circles, dotted line). The shaded region indicates the sizes (SL > 196 SL) for which lionfish THg was significantly higher (α = 0.05) in SEF than in NEF.

THg varied significantly among tissues (F1.2, 66.7 = 9.36, n = 78, p = 0.002), but did not vary between sexes (F1,53 = 1.81, n = 78, p = 0.18; Fig 4), nor was an interaction present. For both sexes, muscle had significantly higher THg than other tissues (muscle > liver > adipose; Dunn-Šidák, Fig 4). In females, THg in ovaries was significantly different from muscle and adipose, but not liver (Dunn-Šidák, Fig 4). THg in all tissues from females were highly correlated (r from 0.66 to 0.88, p < 0.001, Fig 5A–5F); however only THg in muscle and liver tissues were correlated in male lionfish (r = 0.95, p < 0.001, Fig 5I).

Fig 4. Comparison of THg concentrations in lionfish tissues by sex.

Fig 4

Total mercury concentrations in lionfish tissues in female (solid black bars) and male (solid grey bars) lionfish. Male testes were not sampled (nd). No significant differences were observed for tissues as a function of sex; letters (capitals = females; lower case = males) above tissues represent significant differences from total mercury in tissue types from a Ryan’s Q post-hoc test (see text for details) following ANCOVA.

Fig 5. Relationships in THg concentrations among lionfish tissues.

Fig 5

Relationship among THg (all axes are shown in μg g-1 dw) in various tissues in female (Panels A-F, n = 31) and male lionfish (Panels G-I, n = 26). Coefficients from Pearson canonical correlation analysis (r) are inset within each Panel, significant correlations at α = 0.05 with sequential Bonferroni correction are noted by asterisks (**).

Discussion

The main findings of this study were: (1) mercury concentrations in invasive lionfish from northeast and southeast FL were similar to previously reported values and generally below comparable values for commonly consumed reef species, (2) muscle had significantly higher levels of mercury than other tissues or organs, (3) mercury levels increased with size and rates of accumulation differed by location, (4) females were generally higher in mercury than males for a given size, and (5) mercury concentrations in all lionfish tissues examined in this study were highly correlated for females, but only between liver and muscle for males. Collectively, our findings indicate that THg in lionfish is low and lionfish are safe for human consumption.

THg in lionfish from northeast and southeast FL

Mean THg in lionfish (0.05–0.06 μg g-1 ww; Table 1) in our study were within the range previously reported from Florida (0.02–0.15 μg g-1 ww; [52, 53]). Our results were also within the range of lionfish THg studied in Jamaica (0.016–0.061 μg g-1 ww; [51]) and Curaçao (Range: 0.008–0.106 μg g-1 ww; [54]).

Overall, THg in lionfish was lower than native predatory reef fishes that occupy similar trophic positions in offshore hard bottom habitats like those sampled in our study. THg in carnivorous fishes of similar size (e.g., red grouper, Epinephelus morio; black sea bass, Centropristis striata) are 3–4 times higher than the values observed in our study [37, 64] and more than 8–10 times higher in larger, long-lived species such as gag (Mycteroperca microlepis) and red snapper (Lutjanus campechanus, [64, 65]. The presence of low THg in lionfish is somewhat surprising based on studies of diet [9, 19, 23, 66] which indicate that lionfish are carnivores throughout their life history and almost exclusively piscivorous as adults. Indeed, stable isotope analysis paired with diet analyses suggests that both small (presumably young) and larger lionfish are feeding at the same trophic level [54]. Food web modeling and stable isotope analysis also demonstrates a high trophic position for lionfish within the invaded food web and a large degree of overlap with native top predators [24, 25, 67]. Thus, low levels of THg in lionfish do not appear to be related to differences in feeding ecology as has been demonstrated in groupers and sea basses from this region [65]. Low levels of THg in lionfish could reflect species-specific physiological attributes that favor either reduced uptake or increased rates of elimination. Although lionfish physiology is poorly understood; these rates are highly variable across taxa [68] and in fishes [69].

Low THg in lionfish may be at least partially explained by growth dilution, given the extremely rapid growth rates of lionfish relative to native predators [64]. Lionfish grow to nearly 300 mm TL by age two [27, 7072]; while important fishery species from Florida waters such as red grouper [73], black sea bass [74], and grey snapper achieve sizes of 200, 127, and 204 mm TL at age two, respectively. As a result, young lionfish (1–3 years old) of similar size to these species have been accumulating THg for as little as half the time. This rationale is supported by empirical studies for many fishes for which age is a better predictor of THg than body size or mass [75, 76].

Relationship between THg, size and location

Because THg is not readily depurated, it accumulates within tissues over time and the amount of THg in fish tissues is expected to increase as fish age [65, 77]. Our data are in close agreement with this commonly observed pattern; THg in lionfish was significantly positively correlated with fish length (a proxy for age) accounting for between 50 and 60% of the variation (r2 = 0.55–0.61) in THg depending on sex and location of capture (Fig 2A and 2B). Rates of THg bioaccumulation were site-specific and significantly higher in SEF than in NEF resulting in significantly mercury levels for the largest fish (SL > 196mm; Table 2D, Fig 3). Higher concentrations of THg in SEF could be linked to ambient oceanographic conditions such as warmer ocean temperatures, which have been shown to increase methylation rates by marine primary producers [78]. Previous work has noted much larger differences among regions in Florida [53]; however that study examined mercury at a larger spatial scale, in both nearshore and offshore sites, and sampled in locations with both historic point sources (medical waste incineration) and biogeochemical factors that result in enhanced mercury methylation and bioavailability [7981] distinct from our offshore collection sites. In particular, lionfish from the Florida Keys have been shown to be high in mercury [53], a pattern consistent with elevated mercury in other studies [52] and in other piscivorous fishes from that region [37, 41, 82].

Both the positive relationship between THg and size in lionfish and site-specific bioaccumulation rates have been reported previously [53], but the strengths of the relationships were weaker and more variable (r2 = 0.07–0.35) than reported here. The reduced variability observed in our study may result from our regional spatial scale and similar habitats (offshore hard bottom) in comparison with the earlier study which sampled a larger geographic area, at nearshore and offshore locations, and sampled in the Florida Keys which has unique biogeochemical properties due to the nearby Everglades watershed [53, 83].

Comparison of THg in lionfish tissues

The differential uptake and accumulation of THg within the body tissues of fishes is not well resolved. While numerous studies have reported low levels of THg in muscle relative to lipophilic tissues such as liver [84], others have found no difference [85, 86] or elevated THg in muscle [85, 8789]. In this study, THg in lionfish muscle was significantly higher than in other tissues. This finding is in agreement with earlier work [9092] which supports a general pattern of higher THg in muscle relative to internal organs in fish from lightly contaminated localities which is likely the case with our fish collected in offshore marine waters (>20 km from land). A key limitation of the current study was that methylmercury was not quantified. While numerous studies indicate that THg is a good proxy for methylmercury in fish muscle [4648, 55], the fraction of the THg pool present as methylmercury is highly variable in other tissue types [58], can vary as a function of fish size [57] and among species [47, 93]. Future work on Hg speciation in lionfish organ tissues is needed to assess the value of using lionfish as an indicator species to assess ecosystem health and risk (e.g., [91]), and for better understanding ecophysiological mechanisms underlying Hg distribution, detoxification, and sequestration in this species [94]. Despite these limitations, THg represents the maximum possible concentration for methylmercury and observed THg in all tissues were below accepted thresholds for negative health effects [95] and reproductive impairment [96]. Thus, lionfish appear unlikely to be substantially affected by mercury toxicity.

THg in lionfish were generally positively correlated among tissues as is commonly seen in fishes [94]. However, while this relationship was particularly strong for females; only liver and muscle tissue were found to be correlated in males. This finding could indeed reflect real sex-specific differences in physiology leading to differential accumulation and sequestration of THg in fat tissue; however further study will be required to determine if this trend is real and, if so, the physiological processes that underlie it.

Implications for human health

In the present study, THg in lionfish captured from the east coast of Florida were low (0.05 μg g-1), placing them in Florida’s least restrictive consumption advisory level [59] and within the range of fishes (e.g., salmon, tilapia, cod) promoted for safe consumption by the EPA-FDA [44]. Because lionfish are a marine species found in the southeastern U.S. and Caribbean, they are most likely to replace similar regional species such as grouper and snapper in the diet. THg in lionfish is lower than other commonly consumed reef fishes of similar size such as red grouper, (Epinephelus morio, 0.17 μg g-1 ww), grey snapper (Lutjanus griseus, 0.18–0.21 μg g-1 ww), graysby (Cephalopholis cruentata, 0.16 μg g-1 ww), and black sea bass (Centropristis striatus, 0.14 μg g-1 ww; [37, 65] and much lower than fishery legal-sized individuals of larger species such as gag (Mycteroperca microlepis, 0.40 μg g-1 ww), black grouper (M. bonaci, 0.91 μg g-1 ww, and red snapper (L. campechanus, 0.49 μg g-1 ww; [64, 65]. Thus, lionfish would appear to represent a low mercury alternative to these species, many of which have been severely depleted by commercial and recreational fishing pressure.

THg in lionfish increased with size, a general pattern consistently observed in fishes. One of the largest fish (SL > 300) in our study had THg concentrations that would fall into the FDOH limited consumption (0.5–1.5 ppm) category which calls for restricting consumption to once a month for women of childbearing age and children and weekly for all others [59]. However, these large fish are exceptionally rare; lionfish harvested in Florida are predominantly young fish (SL < 250 mm); larger fish (SL > 250) comprised only 5% of individuals in northeast Florida [71].

Overall, lionfish yields a comparable amount of flesh to similar-sized marine food fishes, has high levels of fatty acids beneficial for health, and fares favorably in direct comparisons with other high value marine species [97]. Our findings indicate that levels of THg in lionfish are low and lionfish are safe for human consumption. As such, concerns over THg in lionfish should not present a significant roadblock to the continued development of directed commercial and recreational fisheries for this invasive species.

Acknowledgments

We would like to thank the recreational and commercial spearfishers who collected the lionfish without whom this study would not have been possible. Assistance with sample collection and processing was given by numerous undergraduate and graduate students, in particular Mickhale Green and Corey Corrick. We also thank Chris Guppenberger for initial setup and ongoing technical assistance with the DMA-80 mercury analyzer, and Dr. Amy Lane for creating the liquid mercury standard solutions for calibrations. Samantha Ehnert provided extensive training to numerous undergraduate students and helped to maintain and troubleshoot the DMA-80 over the course of the study duration.

Data Availability

All relevant data files are available from the Dryad database under the Total mercury concentrations of Florida lionfish project at https://doi.org/10.5061/dryad.p8cz8w9r8.

Funding Statement

This work was supported by grants from the National Science Foundation Research Experiences for Undergraduates Program (OCE-1156659; REU - For Students | NSF - National Science Foundation) to JG, the West Marine - Marine Conservation Grant Program (BlueFuture - Grants | West Marine) and University of North Florida Dean’s Leadership Council Faculty Fellowship Program (UNF - College of Arts & Sciences - Dean's Leadership Council Faculty Fellowship Awards) to EGJ, the UNF Department of Biology (UNF - COAS: Biology - Biology Home) and UNF Coastal Biology Flagship Program (UNF - COAS: Biology - about) to AD, and the Guy Harvey Ocean Foundation to MKS.  The results and interpretations of this work are those of the authors and do not represent the official views of the NSF, West Marine, UNF or GHOF. The funders had and will not have a role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Decision Letter 0

Myra E Finkelstein

8 Jul 2020

PONE-D-20-15978

Total mercury concentrations in invasive lionfish (Pterois volitans/miles) from the Atlantic coast of Florida

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The Gateway to Astronaut Photography of Earth (public domain): http://eol.jsc.nasa.gov/sseop/clickmap/

Maps at the CIA (public domain): https://www.cia.gov/library/publications/the-world-factbook/index.html and https://www.cia.gov/library/publications/cia-maps-publications/index.html

NASA Earth Observatory (public domain): http://earthobservatory.nasa.gov/

Landsat: http://landsat.visibleearth.nasa.gov/

USGS EROS (Earth Resources Observatory and Science (EROS) Center) (public domain): http://eros.usgs.gov/#

Natural Earth (public domain): http://www.naturalearthdata.com/

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

Reviewer #3: Yes

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: Yes

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: This manuscript describes the concentrations of total mercury in lionfish from 2 locations off the Atlantic coast of Florida.

The study appears to be well executed, but is very narrow in its conclusions, and does not add any new insights to the scientific knowledge about mercury accumulation in fish species. The results are relevant for the developing market for this species, confirming what others have published already. But unfortunately measuring only total Hg does not allow for new insights into the disposition of mercury. And no attention is paid to possible sources of Hg that may explain the observed regional differences. In the introduction broad sweeping objectives are presented for this project, but almost none of those are addressed in the results and discussion.

Some detailed comments:

Line 52: a more accurate statement is that in the SE USA there is a species continuum for these two species, or even just one hybrid species (see Wilcox CL, Motomura H, Matsunuma M, Bowen BW. Phylogeography of Lionfishes (Pterois) Indicate Taxonomic Over Splitting and Hybrid Origin of the Invasive Pterois volitans. J Hered. 2018 Feb 14;109(2):162-175). Based on my own observations it is difficult to separate the two species on morphological characteristics, and in one location individuals with characteristics of both species can be found.

Line 59: recently a disease outbreak has shown that the species is not all that resistant (Precipitous Declines in Northern Gulf of Mexico Invasive Lionfish Populations Following the Emergence of an Ulcerative Skin Disease. Harris HE, Fogg AQ, Allen MS, Ahrens RNM, Patterson WF 3rd.Sci Rep. 2020 Feb 4;10(1):1934. doi: 10.1038/s41598-020-58886-8)

Line 65/66: However, others have found no significant effects of lionfish on native fish populations (Invasive lionfish had no measurable effect on prey fish community structure across the Belizean Barrier Reef. Hackerott S, Valdivia A, Cox CE, Silbiger NJ, Bruno JF. PeerJ. 2017 May 25;5:e3270. doi: 10.7717/peerj.3270. eCollection 2017.)

Line 72: however, it is doubtful that spear fish derbies will have a significant effect on populations size (Effectiveness of removals of the invasive lionfish: how many dives are needed to deplete a reef? Usseglio P, Selwyn JD, Downey-Wall AM, Hogan JD. PeerJ. 2017 Feb 23;5:e3043. doi: 0.7717/peerj.3043. eCollection 2017)

Line 87: I would say “higher trophic levels” because lionfish are not top predators like sharks, and there food is in general smaller fish.

Line 101 and further: these numbered aims are laudable, but they are hardly at all discussed in the discussion. For instance: there are several papers on lionfish as biomonitoring species for other chemicals, but none of them is discussed. Aims 2 and 3 are also not discussed, only 4.

Line 111: Do you really think that Hg in fish is linked to “high coastal human population density”? For as far as I know, Hg sources along the East Coast are mostly atmospheric deposition of elemental Hg, getting in the atmosphere through coal-fired power plants. In coastal anoxic marsh sediments this elemental Hg is turned into organic Hg, which is bioavailable. In the discussion it is suggested that the Everglades are a source of Hg in fish in the Keys, which makes sense to me. But why would atmospheric deposition in North Florida be different from the South? Where are the coal fired power plants located in FL?

Line 148: I don’t think you need to state that only females have ovaries

Line 236/237: if you set your significance level a 5%, and there are no significant differences between the sexes, then don’t say that the females are generally higher because it is not significant!

Line 284: point 4) see above: if your statistical analysis says there are no differences, then there are no differences!

Line 316-323: this is called growth dilution

Line 337/338: it would be nice to just mention here what these “known processes” are!

Line 363: I fully agree with this statement!

Reviewer #2: *3. Have the authors made all data underlying the findings in their manuscript fully available?

I only said no because on the PDF I received the statement as to where the data can be found is incomplete: "All data files are available from the University of North Florida database (accession number(s) XXX, XXX.)" X's need to be replace with numbers for reader to be able to access. (Maybe they will be on publication, if so, disregard this comment.)

4. Is the manuscript presented in an intelligible fashion and written in standard English?

I answered 'yes' because the edits are minimal. Please see the following comments broken down line by line for typos to fix and minor edits for sentence clarity.

Comments to author:

I realized a bit too late that I wasn’t supposed to provide copyediting along with other review. I am still going to share some details I noticed because I hope they will be useful in assembling the final manuscript. I would like to see this paper published so that others can access these results for comparison and for public knowledge. I appreciate the useful impacts of this study for invasive species control, human health knowledge, and for promoting the idea that lionfish could be consumed instead of other imperiled species.

Line 54 comma after Hamner et al.

Line 63 add comma after Johnston et al.

Line 64 add comma after Cote et al.

Line 65 add comma after Peake et al.

Line 68 add comma after Chagaris et al.

Line 75 add comma after Dahl et al.

Line 88-92 The accumulation of mercury can directly impact fish health (Adams et al., 2010) and can adversely affect many aspects of reproduction (Weis, 2009). Mercury also poses a serious exposure risk for humans who consume fish, particularly for young or pregnant individuals.

First off, I would like to compliment your choice of a gender-neutral term when referring to pregnant people. Thank you! I am only picking at this sentence because the first two times I read through it, it felt moderately unclear whether you are referring to young or pregnant fish or young or pregnant humans. I broke the sentence into two parts as a proposed edit, though if you are very attached to keeping it as one long sentence, I would consider switching the words ‘individual’ and ‘human’ in their placement in the sentence. I know what you mean, but someone unfamiliar with this topic could get the impression that consuming young or pregnant fish carries the greatest health risk.

Line 93 “organic methylmercury (MeHg) (Bloom, 1992)”

Line 94 after “most bioactive” may consider citing a source here or if also from Bloom, 1992, just cite this at the end of the sentence

Line 108 “none of these occurred in our study area” leads the reader to wonder “these what? (characteristics observed or something else)?” upon rereading I realized the intended meaning may have been “none of these studies was conducted in our study area” (which is what your study is uniquely contributing, so you’ll want to make that clear to emphasize its value)

Line 108 Proposed change: Previous studies, …although none of these studies were conducted in our region. End sentence. New sentence: “Only one of the aforementioned studies examined Hg in lionfish across the entire range of sizes currently being harvested” (cite this one alone so readers can find it and compare).

Line 110 “spatial coverage to include an (not and) assessment of Hg in lionfish”

Line 114 “Impacts on (lionfish) health and reproduction.” For the lionfish or the people who eat them could be made clear.

Line 144 “and not available for some tissue samples collected in the field”

What do you mean by this? As in, some fish were not retrieved and brought back to the lab but instead tissue was sampled from them without removal of the whole fish? If you keep this sentence, you may want to elaborate. The other text of your method section implies that all fish were captured whole during the derby. If this is not the case, I would consider adding detail about what other types of tissue may have been collected and how.

Line 146 Good detail! Left side of fish fillet used to represent edible portion. Excellent to include as would help others who wanted to conduct a similar study for their region do so with same methodology. Also good because readers know the part of fish measured is the part people consume.

Line 149 How did you know the samples were completely dry after 48 hours? Constant weight? You have previously established that this drying procedure is effective? May want to tell the reader why you have confidence in this methodology.

Line 154 comma after; Joiris, 2000;

Line 185 I am curious why you chose to use CRM, NIST Standard Ref 2709a San Joaquin soil as a standard reference material. The other QC/QA procedures all seemed totally reasonable for the DMA-80 calibration and it sounds like you had good reproduceability. My only concern would be if you had poor recovery from fish tissue (while perfectly fine recovery from soil) you would not know this from the soil standard or from agreeable replicates of fish tissue (in the event they both had similarly poor recovery).

I do not think this renders the study unpublishable, just wondering why you did not choose a tissue CRM. If your lab previously demonstrated excellent agreement between a fish tissue standard and the soil standard in use, for example, I would consider providing a figure (like a 1:1 plot) to illustrate that fact as part of supplemental data to dispel any reader skepticism regarding the validity of the measurements.

Line 229 “was independent of fish size, sex (add comma) and location.”

Line 261 concentrations is misspelled (a and r are switched)

Line 291-294 “Mean THg in lionfish (0.05 – 0.06 μg g-1 ww; Table 1) in our study were within the range previously reported from Florida (0.02 – 0.15 μg g-1 ww; Huge et al., 2013; Tremain and O’Donnell, 2014).

I would recommend ending that sentence and starting a new one with something to the effect of: Our study also found mean THg in lionfish to be similar to (or greater than) those studied in Jamaica (0.037 μg g-1 ww; Hoo Fung et al., 2013) and Curaçao (Range: 0.008 – 0.016 μg g-1 ww; Ritger et al. 2018).

I suggest this because 1) 0.037 is not a range and 2) 0.05 – 0.06 is not in the range of 0.008-0.016. To set up the sentence as it currently stands just makes me wonder if the other numbers are typos or if the range is missing for Jamaica (as your stated values are also greater than this).

Line 299 change “more the” to “more than 8-10 times higher”

Line 359-360 “this finding is (in) agreement” ‘in’ is missing

Line 366 “can vary as a function (of) fish size” ‘of’ is missing

Line 371 distribution, detoxification, (and) sequestration in this species

Line 381 “artifact of the data” If you keep this statement, I would elaborate on what you mean specifically.

The way it reads currently led me to have the following questions:

Do male fish have less adipose tissue than female fish and did this pose sampling challenges in regards to uniformity?

Is male fish body fat distribution in a different pattern than for females or is it possible that males burn off and replace body fat throughout their lifetimes, therefore disconnecting the THg stored in adipose tissue from the fish’s other tissues? Perhaps female fish adipose tracks well with liver and muscle because all three increase with fish age whereas male fish have a disconnect here?

I understand that all of these questions are beyond the scope of your current study (which you allude to in the next sentence that calls for more research). However, it might be good to give a specific example of a feature that could have led to a data artifact, such as sample size or a particular limitation of the tissue sampling method.

Line 432 and 435 you are using a comma between 2 authors

Line 435 extra space after Sonne C.

Line 450 you use ‘and’ between 2 authors

I would pick one format and use it throughout.

Line 453 and 456 have comma between 2 authors

Line 468 and Line 485 no comma between single author and date

Line 488 you use a comma after Buddo, DSA. Pick one style for this also.

Line 501 comma missing after Sturup S

Line 506 Period missing after USA

Line 512 subtropical does not need to be capitalized if tropical is not

Line 517 “Enviorn” need to become “Environ”

Line 525 Is Mexico intentionally italicized?

Line 529-530 “2016” is listed prior to the journal title (unlike other entries)

Line 526 add “-“ between page numbers

Line 533 period is missing after page number range

Line 549 “966” not justified like others

Line 553 period missing after 345

Line 564 here the date needs to move to line 566

Line 576 here you list all authors whereas other references are listed as et al. after the first 6 names…I would make all references complete like this one

Line 579 list page range, same format as others

Lines 595-604 text is grey rather than black

Line 596 period missing after 22-32

Line 595 and 598 the year is not formatted like other entries

Line 604 is (8) a volume or issue number?

Line 628 and 632 the year is not formatted like others

Line 649 period missing after page range

Line 668 missing dash in page range

Line 744 the word Ranges does not need to be capitalized

Line 747 put the '1' in parentheses like other citations

Line 755 and 785 you used comma after author last name before initials

Line 778 no comma used between last name and initials …pick one style

Line 790 period missing after page numbers

Reviewer #3: I found the study itself to be interesting and novel. It assesses whether lionfish can be safely consumed by people as a food source, but also as a way to cull the population, since they are invasive and ecologically harmful. I found the study design to be appropriate and scientifically sound and the methods (both analytical and statistical) were appropriate and rigorous.

**********

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: Jeannette Isabella Calvin

Reviewer #3: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

PLoS One. 2021 Sep 21;16(9):e0234534. doi: 10.1371/journal.pone.0234534.r002

Author response to Decision Letter 0


8 Jan 2021

Additional Editor Comments:

In your revision please make sure to fully address each of the reviewers' comments.

In particular for reviewer 1, make sure to address this general criticism of the paper:

"But unfortunately measuring only total Hg does not allow for new insights into the disposition of mercury. And no attention is paid to possible sources of Hg that may explain the observed regional differences. In the introduction broad sweeping objectives are presented for this project, but almost none of those are addressed in the results and discussion." and for Reviewer 2 please indicate why a fish tissue SRM was not used.

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

Response: The formatting on the revised manuscript were revised to meet PLOS ONE’s style requirements.

2. In your Methods section, please provide additional location information of the study sites, including geographic coordinates for the data set if available.

Response: The study sites shown in Fig 1 are approximate, but representative, locations of the general areas in which lionfish were captured. The study utilized recreational divers participating in day long lionfish tournaments to collect lionfish and specific locations of capture (i.e., lat, long) was not provided.

3. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide.

Response: The relevant accession numbers or DOIs necessary to access the data will be provided following acceptance.

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We require you to either (1) present written permission from the copyright holder to publish these figures specifically under the CC BY 4.0 license, or (2) remove the figures from your submission:

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In the figure caption of the copyrighted figure, please include the following text: “Reprinted from [ref] under a CC BY license, with permission from [name of publisher], original copyright [original copyright year].”

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The following resources for replacing copyrighted map figures may be helpful:

USGS National Map Viewer (public domain): http://viewer.nationalmap.gov/viewer/

The Gateway to Astronaut Photography of Earth (public domain): http://eol.jsc.nasa.gov/sseop/clickmap/

Maps at the CIA (public domain): https://www.cia.gov/library/publications/the-world-factbook/index.html and https://www.cia.gov/library/publications/cia-maps-publications/index.html

NASA Earth Observatory (public domain): http://earthobservatory.nasa.gov/

Landsat: http://landsat.visibleearth.nasa.gov/

USGS EROS (Earth Resources Observatory and Science (EROS) Center) (public domain): http://eros.usgs.gov/#

Natural Earth (public domain): http://www.naturalearthdata.com/

Response: The original figure which was a modified version of a previously published figure (PeerJ) was deleted. The new figure (Fig 1) was modified from a base map created using the USGS National Map Viewer (public domain): http://viewer.nationalmap.gov/viewer/ as suggested above.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

Reviewer #3: Yes

________________________________________

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

________________________________________

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: Yes

________________________________________

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

________________________________________

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: This manuscript describes the concentrations of total mercury in lionfish from 2 locations off the Atlantic coast of Florida.

The study appears to be well executed, but is very narrow in its conclusions, and does not add any new insights to the scientific knowledge about mercury accumulation in fish species. The results are relevant for the developing market for this species, confirming what others have published already. But unfortunately measuring only total Hg does not allow for new insights into the disposition of mercury. And no attention is paid to possible sources of Hg that may explain the observed regional differences. In the introduction broad sweeping objectives are presented for this project, but almost none of those are addressed in the results and discussion.

Response: Here we provide a summary of revisions to address the reviewers broad overarching comments above (with accompanying highlighted text from the revised manuscript); however detailed responses also follow individual comments reviewer comments below. We have removed the broad sweeping objectives from the revised manuscript and instead focused primarily on THg concentrations in muscle and their potential impact on human health in a developing fishery market for this species.

“The goal of this study was to quantify total mercury (THg) in lionfish as a function of capture location, sex, size and tissue type; information that is critical for evaluating the potential risk to consumers of this species in a rapidly developing fishery.”

THg in this context (human health) is informative as numerous studies demonstrate THg in fish muscle is predominantly methylmercury. Our study expands our knowledge of mercury accumulation in this species by sampling in previously unsampled areas and by increasing the range of sizes (particularly large fish which are most likely to have high mercury) for which information is available.

Previous studies have provided assessments for mercury risk in lionfish (Hoo Fung et al., 2013; Huge et al., 2014; Tremain and O’Donnell, 2014; Ritger et al., 2018); however, none of these studies were conducted in our region of study. Moreover, only one of the aforementioned studies examined mercury in lionfish across the entire range of sizes currently being harvested (Hoo Fung et al., 2013).

The utility of the results of THg in other tissues is more limited, as the reviewer states. For these results, we provide a clear discussion and acknowledgment of the study limitations imposed by measuring only THg.

A key limitation of the current study was that methylmercury was not quantified. While numerous studies indicate that THg is a good proxy for methylmercury in fish muscle (Grieb, 1990; Bloom, 1992; Mieiro et al., 2009; Harley et al. 2015), the fraction of the THg pool present as methylmercury is highly variable in other tissue types (Houserova et al., 2006), can vary as a function of fish size (Joiris et al., 2000) and among species (Mieiro et al., 2009; Berzas Nevado et al., 2011). Future work on Hg speciation in lionfish organ tissues is needed to assess the value of using lionfish as an indicator species to assess ecosystem health and risk (e.g., Havelková et al., 2008), and for better understanding ecophysiological mechanisms underlying Hg distribution, detoxification, and sequestration in this species (Cizdziel et al., 2003). Despite these limitations, THg represents the maximum possible concentration for methylmercury and observed THg in all tissues were below accepted thresholds for negative health effects (Depew et al., 2012) and reproductive impairment (Crump and Trudeau, 2008). Thus, lionfish appear unlikely to be substantially affected by mercury toxicity.

A more detailed discussion of the potential drivers of regional differences is also included the revised manuscript.

Some detailed comments:

Line 52: a more accurate statement is that in the SE USA there is a species continuum for these two species, or even just one hybrid species (see Wilcox CL, Motomura H, Matsunuma M, Bowen BW. Phylogeography of Lionfishes (Pterois) Indicate Taxonomic Over Splitting and Hybrid Origin of the Invasive Pterois volitans. J Hered. 2018 Feb 14;109(2):162-175). Based on my own observations it is difficult to separate the two species on morphological characteristics, and in one location individuals with characteristics of both species can be found.

Response: We have incorporated this suggested change and added the suggested reference.

A marine invader of particular concern in the western Atlantic Ocean is the Indo-Pacific lionfish (a species continuum of two morphologically indistinct species, Pterois miles and P. volitans or potentially a hybrid of the two; Wilcox et al., 2018).

Line 59: recently a disease outbreak has shown that the species is not all that resistant (Precipitous Declines in Northern Gulf of Mexico Invasive Lionfish Populations Following the Emergence of an Ulcerative Skin Disease. Harris HE, Fogg AQ, Allen MS, Ahrens RNM, Patterson WF 3rd.Sci Rep. 2020 Feb 4;10(1):1934. doi: 10.1038/s41598-020-58886-8)

Response: We have incorporated this suggested change and added the suggested reference.

Lionfish have long venomous spines that deter predation by native predators (Mumby et al., 2011) and are resistant to common parasites (Sikkel et al., 2014); however a recently discovered outbreak of an ulcerative skin disease in lionfish in the Gulf of Mexico suggests some susceptibility to disease (Harris et al., 2020).

Line 65/66: However, others have found no significant effects of lionfish on native fish populations (Invasive lionfish had no measurable effect on prey fish community structure across the Belizean Barrier Reef. Hackerott S, Valdivia A, Cox CE, Silbiger NJ, Bruno JF. PeerJ. 2017 May 25;5:e3270. doi: 10.7717/peerj.3270. eCollection 2017.)

Response: We have incorporated this suggested change and added the suggested reference.

Lionfish have been shown to reduce native fish recruitment (Albins and Hixon, 2008) and overall native species biomass (Green et al., 2012) in some studies, but not in others (Hackerott et al., 2017).

Line 72: however, it is doubtful that spear fish derbies will have a significant effect on populations size (Effectiveness of removals of the invasive lionfish: how many dives are needed to deplete a reef? Usseglio P, Selwyn JD, Downey-Wall AM, Hogan JD. PeerJ. 2017 Feb 23;5:e3043. doi: 0.7717/peerj.3043. eCollection 2017)

Response: We have incorporated this suggested change and added the suggested reference.

Such efforts are being actively promoted by management agencies throughout the western Atlantic Ocean and Caribbean Sea, but require careful implementation to be most effective (Usseglio et al. 2017).

Line 87: I would say “higher trophic levels” because lionfish are not top predators like sharks, and there food is in general smaller fish.

Response: We have incorporated this suggested change.

Line 101 and further: these numbered aims are laudable, but they are hardly at all discussed in the discussion. For instance: there are several papers on lionfish as biomonitoring species for other chemicals, but none of them is discussed. Aims 2 and 3 are also not discussed, only 4.

Response: We have incorporated this suggested change to focus on the primary goal of the study.

The goal of this study was to quantify total mercury (THg) in lionfish as a function of capture location, sex, size and tissue type; information that is critical for evaluating the potential risk to consumers of this species in a rapidly developing fishery.

Line 111: Do you really think that Hg in fish is linked to “high coastal human population density”? For as far as I know, Hg sources along the East Coast are mostly atmospheric deposition of elemental Hg, getting in the atmosphere through coal-fired power plants. In coastal anoxic marsh sediments this elemental Hg is turned into organic Hg, which is bioavailable. In the discussion it is suggested that the Everglades are a source of Hg in fish in the Keys, which makes sense to me. But why would atmospheric deposition in North Florida be different from the South? Where are the coal fired power plants located in FL?

Response: We have changed the wording of the sentence to remove confusion to remove the phrase “high coastal human population density”. The rationale for including “high coastal human population density” as a qualifier was not that these areas are likely to have high levels of THg, but that these areas have many people and high levels of recreational fishing effort making the lack of information on THg in these areas important from a public health perspective. The regional differences we observed (SEF vs NEF) and possible explanations are detailed in the discussion (and below).

Line 148: I don’t think you need to state that only females have ovaries

Response: We have incorporated this suggested change.

For a subset of male (n=26) and female fish (n=31), we collected additional samples of liver, adipose and ovarian tissue for analysis.

Line 236/237: if you set your significance level a 5%, and there are no significant differences between the sexes, then don’t say that the females are generally higher because it is not significant!

Response: We believe that the two statements are not incongruous. THg concentrations may be not significantly different between sexes and still generally higher in females than in males for a given size. The difference becomes larger with size, which we believe results from sex-specific differences in growth rates (growth dilution) which are well documented for this species. The statement is not crucial to the manuscript, so I will remove it if the editor concurs with the reviewer.

Line 284: point 4) see above: if your statistical analysis says there are no differences, then there are no differences!

Response: See above. Assuming the most likely difference between groups is 0 following a statistical test is a common problem in interpreting statistical output. The most likely estimate of the difference between the groups is the observed difference (a confidence interval for which will include 0). Again, the statement is not crucial to the manuscript, so I will remove it if the editor concurs with the reviewer.

Line 316-323: this is called growth dilution

Response: We have incorporated this suggested change.

Low THg in lionfish may be at least partially explained by growth dilution, given the extremely rapid growth rates of lionfish relative to native predators (Zapp-Sluis et al., 2013).

Line 337/338: it would be nice to just mention here what these “known processes” are!

Response: We have incorporated more discussion into possible processes causing regional differences in THg.

Higher concentrations of THg in SEF could be linked to ambient oceanographic conditions such as warmer ocean temperatures, which have been shown to increase methylation rates by marine primary producers (Lee and Fisher, 2016). Previous work has noted much larger differences among regions in Florida (Tremain and O’Donnell, 2014); however that study examined mercury at a larger spatial scale, in both nearshore and offshore sites, and sampled in locations with both historic point sources (medical waste incineration) and biogeochemical factors that result in enhanced mercury methylation and bioavailability (Chen et al., 2009; Sparling, 2009; Driscoll et al., 2012) distinct from our offshore collection sites.

Line 363: I fully agree with this statement!

Reviewer #2: *3. Have the authors made all data underlying the findings in their manuscript fully available?

I only said no because on the PDF I received the statement as to where the data can be found is incomplete: "All data files are available from the University of North Florida database (accession number(s) XXX, XXX.)" X's need to be replace with numbers for reader to be able to access. (Maybe they will be on publication, if so, disregard this comment.)

Response: The relevant accession numbers or DOIs necessary to access the data will be provided following acceptance as required by the journal.

4. Is the manuscript presented in an intelligible fashion and written in standard English?

I answered 'yes' because the edits are minimal. Please see the following comments broken down line by line for typos to fix and minor edits for sentence clarity.

Comments to author:

I realized a bit too late that I wasn’t supposed to provide copyediting along with other review. I am still going to share some details I noticed because I hope they will be useful in assembling the final manuscript. I would like to see this paper published so that others can access these results for comparison and for public knowledge. I appreciate the useful impacts of this study for invasive species control, human health knowledge, and for promoting the idea that lionfish could be consumed instead of other imperiled species.

Line 54 comma after Hamner et al.

Line 63 add comma after Johnston et al.

Line 64 add comma after Cote et al.

Line 65 add comma after Peake et al.

Line 68 add comma after Chagaris et al.

Line 75 add comma after Dahl et al.

Response: We have incorporated these suggested edits and appreciate the level of detail with which reviewer 2 edited the manuscript.

Line 88-92 The accumulation of mercury can directly impact fish health (Adams et al., 2010) and can adversely affect many aspects of reproduction (Weis, 2009). Mercury also poses a serious exposure risk for humans who consume fish, particularly for young or pregnant individuals.

First off, I would like to compliment your choice of a gender-neutral term when referring to pregnant people. Thank you! I am only picking at this sentence because the first two times I read through it, it felt moderately unclear whether you are referring to young or pregnant fish or young or pregnant humans. I broke the sentence into two parts as a proposed edit, though if you are very attached to keeping it as one long sentence, I would consider switching the words ‘individual’ and ‘human’ in their placement in the sentence. I know what you mean, but someone unfamiliar with this topic could get the impression that consuming young or pregnant fish carries the greatest health risk.

Response: We have incorporated this suggested edit.

The accumulation of mercury can not only directly impact fish health (Adams et al., 2010) and adversely affect many aspects of reproduction (Weis, 2009). Mercury also poses a serious exposure risk for humans who consume fish, particularly for young or pregnant individuals (NRC, 2000; U.S. EPA, 2001; Karagas et al., 2012).

Line 93 “organic methylmercury (MeHg) (Bloom, 1992)”

Line 94 after “most bioactive” may consider citing a source here or if also from Bloom, 1992, just cite this at the end of the sentence

Response: We have incorporated this suggested edit.

Further, most of the mercury present in fish muscle is present as organic methylmercury (MeHg) (Bloom, 1992; Mieiro et al., 2009; Harley et al. 2015), the most highly toxic and bioactive form (Mergler et al., 2007).

Line 108 “none of these occurred in our study area” leads the reader to wonder “these what? (characteristics observed or something else)?” upon rereading I realized the intended meaning may have been “none of these studies was conducted in our study area” (which is what your study is uniquely contributing, so you’ll want to make that clear to emphasize its value)

Response: We changed the text for clarification.

Previous studies have provided assessments for mercury risk in lionfish (Hoo Fung et al., 2013; Huge et al., 2014; Tremain and O’Donnell, 2014; Ritger et al., 2018); however, none of these studies were conducted in our region of study.

Line 108 Proposed change: Previous studies, …although none of these studies were conducted in our region. End sentence. New sentence: “Only one of the aforementioned studies examined Hg in lionfish across the entire range of sizes currently being harvested” (cite this one alone so readers can find it and compare).

Response: We have incorporated this suggested edit.

Previous studies have provided assessments for mercury risk in lionfish (Hoo Fung et al., 2013; Huge et al., 2014; Tremain and O’Donnell, 2014; Ritger et al., 2018); however, none of these studies were conducted in our region of study. Moreover, only one of the aforementioned studies examined mercury in lionfish across the entire range of sizes currently being harvested (Hoo Fung et al., 2013).

Line 110 “spatial coverage to include an (not and) assessment of Hg in lionfish”

Response: We have incorporated this suggested edit.

Line 114 “Impacts on (lionfish) health and reproduction.” For the lionfish or the people who eat them could be made clear.

Response: We changed the text for clarification.

This study builds on earlier work by (1) expanding the spatial coverage to include an assessment of mercury in lionfish from unstudied regions, (2) expanding the range of sizes examined (particularly large individuals which have the highest potential for mercury bioaccumulation), and (3) quantifying mercury levels in different lionfish tissues to assess potential impacts on lionfish health and reproduction

Line 144 “and not available for some tissue samples collected in the field”

What do you mean by this? As in, some fish were not retrieved and brought back to the lab but instead tissue was sampled from them without removal of the whole fish? If you keep this sentence, you may want to elaborate. The other text of your method section implies that all fish were captured whole during the derby. If this is not the case, I would consider adding detail about what other types of tissue may have been collected and how.

Response: We changed the text to add more detail.

Sex determination was not possible for many smaller immature individuals (Morris et al., 2012) and not available for a limited number of lionfish (n=5) which sustained extensive damage during capture and field processing.

Line 146 Good detail! Left side of fish fillet used to represent edible portion. Excellent to include as would help others who wanted to conduct a similar study for their region do so with same methodology. Also good because readers know the part of fish measured is the part people consume.

Response: As suggested, no change was made.

Line 149 How did you know the samples were completely dry after 48 hours? Constant weight? You have previously established that this drying procedure is effective? May want to tell the reader why you have confidence in this methodology.

Response: We changed the text to add more detail.

All samples were dried in a 60°C oven for 48 hours (a duration sufficient to achieve constant weight of samples) then homogenized using a mortar and pestle prior to analysis of total mercury concentrations (hereafter THg).

Line 154 comma after; Joiris, 2000;

Response: We have incorporated this suggested edit.

Line 185 I am curious why you chose to use CRM, NIST Standard Ref 2709a San Joaquin soil as a standard reference material. The other QC/QA procedures all seemed totally reasonable for the DMA-80 calibration and it sounds like you had good reproduceability. My only concern would be if you had poor recovery from fish tissue (while perfectly fine recovery from soil) you would not know this from the soil standard or from agreeable replicates of fish tissue (in the event they both had similarly poor recovery).

I do not think this renders the study unpublishable, just wondering why you did not choose a tissue CRM. If your lab previously demonstrated excellent agreement between a fish tissue standard and the soil standard in use, for example, I would consider providing a figure (like a 1:1 plot) to illustrate that fact as part of supplemental data to dispel any reader skepticism regarding the validity of the measurements.

Response: The reviewer raises a valid point. Unfortunately, at the time of the study we attempted to obtain fish tissue CRMs, however they were not available for purchase from vendors. Thus, we purchased the Soil CRM. We have recently acquired fish tissue CRM, but have not yet conducted the suggested analysis.

Line 229 “was independent of fish size, sex (add comma) and location.”

Response: We have incorporated this suggested edit.

Line 261 concentrations is misspelled (a and r are switched)

Response: We have incorporated this suggested edit.

Line 291-294 “Mean THg in lionfish (0.05 – 0.06 μg g-1 ww; Table 1) in our study were within the range previously reported from Florida (0.02 – 0.15 μg g-1 ww; Huge et al., 2013; Tremain and O’Donnell, 2014).

I would recommend ending that sentence and starting a new one with something to the effect of: Our study also found mean THg in lionfish to be similar to (or greater than) those studied in Jamaica (0.037 μg g-1 ww; Hoo Fung et al., 2013) and Curaçao (Range: 0.008 – 0.016 μg g-1 ww; Ritger et al. 2018).

I suggest this because 1) 0.037 is not a range and 2) 0.05 – 0.06 is not in the range of 0.008-0.016. To set up the sentence as it currently stands just makes me wonder if the other numbers are typos or if the range is missing for Jamaica (as your stated values are also greater than this).

Response: We have incorporated this suggested edit to add clarification. There was also a typo: 0.016 was 0.106 for THg in Curacao, which is the range of our results.

Mean THg in lionfish (0.05 – 0.06 μg g-1 ww; Table 1) in our study were within the range previously reported from Florida (0.02 – 0.15 μg g-1 ww; Huge et al., 2013; Tremain and O’Donnell, 2014). Our results were also within the range of lionfish THg studied in Jamaica (0.016 – 0.061 μg g-1 ww; Hoo Fung et al., 2013) and Curaçao (Range: 0.008 – 0.106 μg g-1 ww; Ritger et al. 2018).

Line 299 change “more the” to “more than 8-10 times higher”

Response: We have incorporated this suggested edit.

Line 359-360 “this finding is (in) agreement” ‘in’ is missing

Response: We have incorporated this suggested edit.

Line 366 “can vary as a function (of) fish size” ‘of’ is missing

Response: We have incorporated this suggested edit.

Line 371 distribution, detoxification, (and) sequestration in this species

Response: We have incorporated this suggested edit.

Line 381 “artifact of the data” If you keep this statement, I would elaborate on what you mean specifically.

The way it reads currently led me to have the following questions:

Do male fish have less adipose tissue than female fish and did this pose sampling challenges in regards to uniformity?

Is male fish body fat distribution in a different pattern than for females or is it possible that males burn off and replace body fat throughout their lifetimes, therefore disconnecting the THg stored in adipose tissue from the fish’s other tissues? Perhaps female fish adipose tracks well with liver and muscle because all three increase with fish age whereas male fish have a disconnect here?

I understand that all of these questions are beyond the scope of your current study (which you allude to in the next sentence that calls for more research). However, it might be good to give a specific example of a feature that could have led to a data artifact, such as sample size or a particular limitation of the tissue sampling method.

Response: We have removed this statement as suggested by the reviewer.

Line 432 and 435 you are using a comma between 2 authors

Line 435 extra space after Sonne C.

Line 450 you use ‘and’ between 2 authors

I would pick one format and use it throughout.

Line 453 and 456 have comma between 2 authors

Line 468 and Line 485 no comma between single author and date

Line 488 you use a comma after Buddo, DSA. Pick one style for this also.

Line 501 comma missing after Sturup S

Line 506 Period missing after USA

Line 512 subtropical does not need to be capitalized if tropical is not

Line 517 “Enviorn” need to become “Environ”

Line 525 Is Mexico intentionally italicized?

Line 529-530 “2016” is listed prior to the journal title (unlike other entries)

Line 526 add “-“ between page numbers

Line 533 period is missing after page number range

Line 549 “966” not justified like others

Line 553 period missing after 345

Line 564 here the date needs to move to line 566

Line 576 here you list all authors whereas other references are listed as et al. after the first 6 names…I would make all references complete like this one

Line 579 list page range, same format as others

Lines 595-604 text is grey rather than black

Line 596 period missing after 22-32

Line 595 and 598 the year is not formatted like other entries

Line 604 is (8) a volume or issue number?

Line 628 and 632 the year is not formatted like others

Line 649 period missing after page range

Line 668 missing dash in page range

Line 744 the word Ranges does not need to be capitalized

Line 747 put the '1' in parentheses like other citations

Line 755 and 785 you used comma after author last name before initials

Line 778 no comma used between last name and initials …pick one style

Line 790 period missing after page numbers

Response: We have incorporated each of these suggested edits, and very much appreciate the detail with which reviewer 2 edited the document. These edits were quite helpful in generating the revised manuscript.

Reviewer #3: I found the study itself to be interesting and novel. It assesses whether lionfish can be safely consumed by people as a food source, but also as a way to cull the population, since they are invasive and ecologically harmful. I found the study design to be appropriate and scientifically sound and the methods (both analytical and statistical) were appropriate and rigorous.

Attachment

Submitted filename: Johnson et al - Response to reviewers - Final.docx

Decision Letter 1

Myra E Finkelstein

13 Jan 2021

PONE-D-20-15978R1

Total mercury concentrations in invasive lionfish (Pterois volitans/miles) from the Atlantic coast of Florida

PLOS ONE

Dear Dr. Johnson,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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We look forward to receiving your revised manuscript.

Kind regards,

Myra E Finkelstein

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

One major concern with the QAQC of the mercury data was that the authors used an inappropriate CRM to assess the accuracy of total mercury measurements. This was pointed out by one of the reviewers on the prior draft. In response to this comment the authors stated in this revision: "The reviewer raises a valid point. Unfortunately, at the time of the study we attempted to obtain fish tissue CRMs, however they were not available for purchase from vendors. Thus, we purchased the Soil CRM. We have recently acquired fish tissue CRM, but have not yet conducted the suggested analysis."

As the main findings and conclusions of this paper are based on total mercury measurements, before the revised paper is sent back out to review, I think it is necessary to run the fish tissue CRM that has been purchased (which should be a quick and simple thing to do to) compare with the soil CRM as the reviewer suggested.

Comment from the reviewer on the prior draft: "If your lab previously demonstrated excellent agreement between a fish tissue standard and the soil standard in use, for example, I would consider providing a figure (like a 1:1 plot) to illustrate that fact as part of supplemental data to dispel any reader skepticism regarding the validity of the measurements.”

[Note: HTML markup is below. Please do not edit.]

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PLoS One. 2021 Sep 21;16(9):e0234534. doi: 10.1371/journal.pone.0234534.r004

Author response to Decision Letter 1


8 Jun 2021

This is the second revision of the manuscript. In this second revision, we specifically address reviewer/editor concerns relative to our use of a solid soil Certified Reference Material (CRM) and not a fish tissue CRM. As suggested, we have conducted supplemental paired analysis of Hg using both CRMs. These analyses demonstrated that both our original standard CRM and the fish protein CRM (DORM-4) were within expected ranges. Specifically, average DORM-4 concentrations for Hg were 0.38 ug/g (expected reference range: 0.41 +/- 0.055 ug/g).

Decision Letter 2

Myra E Finkelstein

13 Jul 2021

Total mercury concentrations in invasive lionfish (Pterois volitans/miles) from the Atlantic coast of Florida

PONE-D-20-15978R2

Dear Dr. Johnson,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Myra E Finkelstein

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

**********

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The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

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Reviewer #1: Yes

Reviewer #2: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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6. Review Comments to the Author

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Reviewer #1: (No Response)

Reviewer #2: Excellent! Thank you for including the fish tissue standard information. The sections rewritten for clarity improved the readability of the manuscript. It is now even more informative and valuable due to the inclusion of specific details (i.e. description of historic point sources/historic incineration of medical waste, reason as to why the sex could not be determined for certain individuals).

**********

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Reviewer #1: No

Reviewer #2: Yes: Nettie Calvin

Acceptance letter

Myra E Finkelstein

9 Sep 2021

PONE-D-20-15978R2

Total mercury concentrations in invasive lionfish (Pterois volitans/miles) from the Atlantic coast of Florida

Dear Dr. Johnson:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Myra E Finkelstein

Academic Editor

PLOS ONE

Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    Attachment

    Submitted filename: Johnson et al - Response to reviewers - Final.docx

    Data Availability Statement

    All relevant data files are available from the Dryad database under the Total mercury concentrations of Florida lionfish project at https://doi.org/10.5061/dryad.p8cz8w9r8.


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