Marine Pollution Bulletin: Samantha M. Iliff, Rachel J. Harris, Elizabeth W. Stoner T
Marine Pollution Bulletin: Samantha M. Iliff, Rachel J. Harris, Elizabeth W. Stoner T
Marine Pollution Bulletin: Samantha M. Iliff, Rachel J. Harris, Elizabeth W. Stoner T
Keywords: In recent decades, oyster reefs have been deteriorating throughout North America as a result of multiple in-
Americorophium amphipod teracting anthropogenic stressors, including pesticide pollution. Here we elucidated the potential chronic effects
Carbaryl of the commonly utilized pesticide, carbaryl, on oyster reef communities in the Loxahatchee River Estuary in
Crustacean southeast Florida. Though carbaryl had a limited effect on total epifaunal community diversity, species richness
Pesticide
and evenness, the results of this experiment indicate that carbaryl significantly shifted crustacean community
Loxahatchee River Estuary
Oyster reef
composition, resulting in a substantial loss in total crustacean abundance. One crustacean in particular,
Americorophium spp. (tube building amphipod), was significantly less abundant within the carbaryl treatment,
driving the shift in crustacean community composition. Ultimately, our results signal that pesticide pollution in
estuaries will negatively impact crustaceans. Over time, this may shift benthic community composition, po-
tentially disrupting species interactions and threatening valuable economic and ecosystem services.
⁎
Corresponding author currently at: Department of Ecology and Evolutionary Biology, University of Michigan, 1105 North University Ave., Ann Arbor, MI 48109,
USA
E-mail addresses: siliff@umich.edu (S.M. Iliff), rachel.harris@lrecd.org (R.J. Harris), estoner@bentley.edu (E.W. Stoner).
https://doi.org/10.1016/j.marpolbul.2019.06.060
Received 1 March 2019; Received in revised form 15 June 2019; Accepted 20 June 2019
Available online 10 July 2019
0025-326X/ © 2019 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).
S.M. Iliff, et al. Marine Pollution Bulletin 146 (2019) 502–508
mortality occurred at high concentrations of pesticides (100 μg/L), and plaster blocks containing carbaryl. The carbaryl blocks were made by
exposure to low concentrations (1–10 μg/L) had sub-lethal effects on mixing Plaster of Paris, 80% carbaryl wettable powder and cold water
the crabs, resulting in altered behaviors, decreased muscle function, (Whalen et al., 2013). As a procedural control, additional plaster blocks
and increased reaction time (Schroeder-Spain et al., 2018). As such, were made using the same method without the addition of carbaryl.
pesticide pollution can influence invertebrate populations directly via The blocks were allowed to dry and harden and were then removed
toxicity or indirectly through decreased foraging, or by increasing from their molds and affixed to strings (Fig. 2).
predation risk (Schroeder-Spain, 2017). Although there is a need to To test the effects of carbaryl on oyster reef community composition
understand the effects of acute pesticide toxicity on valuable species, in situ, we designed a field experiment to quantify how carbaryl ex-
few studies have examined the effects of chronic pesticide exposure on posure may influence epifaunal community composition. We tested
estuarine communities. three treatments in this experiment: carbaryl additions (using carbaryl
One common pesticide in the United States that could affect benthic dissolution blocks), a plaster-control treatment (using dissolution
estuarine organisms is carbaryl, a broad-spectrum organocarbamate blocks made without carbaryl), and a double control group with no
(Donaldson et al., 2002). Carbaryl works by inhibiting acet- treatment of dissolution blocks. This double control method was im-
ylcholinesterase in arthropods, causing death by disrupting the nervous plemented to analyze the procedural effects of plaster dissolution on
system (Dumbauld et al., 2001; Relyea, 2005). Commercially available response variables. To simulate the hard-bottom microhabitat of an
as Sevin Insecticide, carbaryl was ranked as the number one most oyster reef, a string of five, 100 cm2 travertine tiles was hung off of T-
commonly used insecticide in the U.S. home and garden market (Grube shape structures constructed with PVC pipes (Fig. 2). We elected to use
et al., 2011). Although insecticides are more commonly used in ter- non-invasive removable tiles rather than a natural oyster reef in order
restrial systems to deter insect predation on plants, carbaryl has also to easily collect and quantify mobile and highly cryptic reef species
been used in intertidal oyster farming practices (Simenstad and Fresh, without removing or damaging a section of natural reef. This method
1995). Since the 1960s oyster growers in Willapa, Bay (Washington accurately simulates an oyster reef, allowing for oyster spat recruitment
State, USA), have treated farmed oyster beds with carbaryl in alter- and settlement (Metz et al., 2015). Each tile was vertically separated by
nating cycles of short-term exposure to protect their stock from bur- a 6.35 mm spacer. The lower tile of each string hung 30 cm above the
rowing thalassinid shrimp (Feldman et al., 2000; Dumbauld et al., sediment in the shallow sub-intertidal zone, and the top-most tile of
2001). Numerous studies have evaluated the lethal and sub-lethal ef- each string was at least 30 cm below the surface at low tide. Each dis-
fects of carbaryl on marine organisms (Estes, 1986; Fulton and Key, solution block was then hung directly adjacent (~3 cm) to a string of
2001; Canty et al., 2007; Schroeder-Spain et al., 2018); however, there recruitment tiles.
is a paucity of data regarding effects of chronic carbaryl exposure in Eighteen recruitment structures were deployed 2 m apart at the field
estuarine ecosystems, especially oyster reefs. It is critical to identify the site. The 2 m structure separation was chosen based on previously es-
impacts of chronic exposure of pesticides because it is likely that pro- tablished methods to locally increase carbaryl concentrations relative to
longed exposure to pesticides could substantially degrade ecosystems controls (see Poore et al., 2009; Cook et al., 2011; and Whalen et al.,
though cascading and multiple-stressor effects. To this end, the goal of 2013). In total, thirty-six strings of recruitment tiles were hung, with
this study was to determine whether chronic carbaryl exposure would one on each end of each T structure. Strings were randomly assigned to
alter oyster reef community composition in a sub-tropical estuarine a treatment, and then assigned to a T structure. Dissolution blocks were
waterbody. In having a better understanding of the effects of carbaryl replaced every 3–5 days and the time and date were recorded. Every
exposure on oyster reefs, we are providing resource managers with a dissolution block was weighed before deployment and after retrieval to
holistic understanding needed to fully assess anthropogenic impacts on measure dissolution rates. Sessile and epibenthic organisms were al-
oyster reefs. lowed to colonize the tiles for 28 days from September 13th–October
11th 2017, which is a time frame that allows for recruitment and set-
2. Methods tlement of eastern oyster (Crassostrea virginica) spat in the Loxahatchee
River estuary (Metz et al., 2015).
2.1. Study area
2.3. Sample collection, processing, and data analysis
We conducted this experiment in the Southwest Fork of the sub-
tropical Loxahatchee River Estuary in Jupiter, Florida (Fig. 1). At this After the 28-day experiment, both sessile and vagile fauna asso-
site, salinities ranged from 0.1 to 34.7 ppt with a mean of 11.2 ppt and ciated with the recruitment tiles were collected by gently enclosing the
water temperatures ranged from 25 to 32 °C (available at: https:// entire string of tiles in a gallon bag before removing the T structures. All
loxahatcheeriver.org/river/river-keeper/), providing suitable oyster unattached organic material was rinsed off of the tiles and strained
habitat requirement (Barnes et al., 2007). The Loxahatchee River Es- through a 500 μm sieve. All strained material was preserved in 70%
tuary covers roughly 4 km2, drains a watershed of approximately ethanol and stained with a 10% Rose Bengal dye solution. All of the
700 km2 and flows into the Atlantic Ocean at the Jupiter Inlet (Ridler epifauna were then identified to the lowest taxonomic level practical,
et al., 2006). The watershed covers a variety of habitats including riv- quantified, and recorded. After the tiles had been rinsed, they were
erine wetland and cypress communities, and estuarine mangroves, oy- allowed to dry overnight. A dissecting microscope was used to analyze
ster reefs, and seagrass beds. Although the system is currently flow and enumerate all sessile fauna on the middle three tiles as outlined in
managed, the combination of altered freshwater inputs and deterior- Haven and Fritz (1985).
ating water quality (see Stoner and Arrington, 2017) have driven shifts The initial and final weights of the dissolution blocks, as well as the
in oyster bed community composition and ecosystem function (Van date and time of deployment and collection, were then used to de-
Arman et al., 2005; Roberts et al., 2008; Parker et al., 2013; Jud, 2014), termine the rate of dissolution of blocks (Thompson and Glenn, 1994)
emphasizing a need for effective resource management and restoration. and dissolution of carbaryl (80% by dry weight). The following equa-
Although water quality is regularly monitored in the Loxahatchee River tion was used where D is dissolution rate, Wi is initial weight, Wf is final
Estuary, monitoring for pesticides has been limited. weight, and θ is total time (in days) dissolving:
Wi Wf
2.2. Experimental design D=
In order to test the effects of carbaryl on an estuarine oyster reef, we Dcarbaryl = D × 0.8 × 0.10
used a method developed by Poore et al. (2009) using slow-dissolving
503
S.M. Iliff, et al. Marine Pollution Bulletin 146 (2019) 502–508
Fig. 1. Map of the study site, indicated with a bullseye, within the Loxahatchee River Estuary, Jupiter, FL.
When comparing overall mean density between treatments, each (set at 999 unique permutations) were used to identify significant dif-
tile string was treated as a discrete sample in which mean standardized ferences between treatments and similarity percentages (SIMPER) were
faunal densities were calculated across the middle three tiles for each used to verify the individual species driving community differences
tile string. One-way ANOVAs were used to test for difference in in- between treatments (Anderson, 2001). Community composition ana-
dividual species densities between treatments, and the non-parametric lysis was conducted using Primer PERMANOVA+v7.0 and all other
test, Kruskal-Wallis test, was used when unequal variance was identi- analyses using with SPSS v24 statistical software.
fied. p-Values ≤ 0.05 were considered statistically significant, while p-
values between 0.05 and 0.10 were considered marginally significant. 3. Results
Community composition dissimilarities were analyzed on a com-
munity wide basis (total fauna) and within the crustacean community. The initial and final weights of the dissolution blocks, as well as the
Multidimensional scaling (MDS) plots were used to visualize the simi- date and time of deployment and collection, were used to determine the
larity and dissimilarity of macrofaunal community compositions (based rate of dissolution of blocks as well as dissolution of carbaryl. When the
on a Bray-Curtis resemblance matrix) by carbaryl, plaster, and control carbaryl dissolution blocks were deployed, each contained approxi-
treatments. The vectors on the MDS plots (set at a Pearson's R Value of mately 15 g of carbaryl by dry weight and ended with approximately
0.75) were used to identify which species were driving the similarities 8 g of carbaryl by dry weight, with an average carbaryl dissolution rate
within and dissimilarity between treatments. Permutational Analysis of of 2.64 ± 0.3 g/day; the average dissolution of carbaryl blocks and
Variance (PERMANOVA) and post-hoc pair-wise PERMANOVA tests plaster blocks was quite similar at 32.9 g/day and 30.0 g/day,
504
S.M. Iliff, et al. Marine Pollution Bulletin 146 (2019) 502–508
Table 1
Mean abundance (individuals/m2) ± standard deviation of the most-abundant animals found on recruitment tiles between treatments. Species are listed by clas-
sification and order of abundance within the control treatment. * indicates marginal difference between treatments (ANOVA or Kruskal-Wallis, 0.05 < p < 0.10).
** indicates significant difference between treatments (ANOVA or Kruskal-Wallis, p < 0.05).
Scientific name Common name Control Plaster Carbaryl
Mollusca
C. virginica Eastern oyster 18,560 ± 1506 19,440 ± 4343 20,450 ± 7321
G. demissa Ribbed mussel 1450 ± 633 1060 ± 396 1425 ± 608
Neritina spp. Nerite snail 118 ± 84 152 ± 104 76 ± 39
Arthropoda
**Americorophium spp. Tube building amphipod 4392 ± 3197 2406 ± 765 998 ± 175
*B. eburneus Ivory barnacle 4960 ± 1306 8170 ± 3366 5616 ± 826
H. laguna Amphipod 2952 ± 589 3026 ± 1263 2708 ± 1082
Insecta Insect larvae 2182 ± 1122 1814 ± 2293 1084 ± 760
S. stanfordi Amphipod 1574 ± 740 1402 ± 1059 1540 ± 704
B. amphitrite Purple striped barnacle 976 ± 625 2273 ± 1518 1670 ± 215
*Isopoda Isopod 176 ± 65 190 ± 45 110 ± 49
Other
Membranipora spp. Encrusting Bryozoan 440 ± 203 403 ± 130 496 ± 215
Nereididae Nereid worms 74 ± 30 80 ± 70 68 ± 33
Hydroides sp.a Serpulid worm 55.6% 50.4% 50.6%
Notes: Sessile organisms were quantified on the tile surfaces from the middle three tiles totaling an area of 0.06 m2. The vagile organisms were rinsed off the entire
string of tiles from an area of 0.1 m2.
a
Serpulid worms were quantified using percent cover because densities often exceeded 50,000 individuals/m2.
505
S.M. Iliff, et al. Marine Pollution Bulletin 146 (2019) 502–508
4. Discussion
506
S.M. Iliff, et al. Marine Pollution Bulletin 146 (2019) 502–508
Within oyster reefs, crustaceans are among the most abundant taxa, Pesticides such as carbaryl have the potential to significantly alter
second only to bivalves (Bahr Jr, 1976; Bahr and Lanier, 1981, Wells, crustacean communities, which would have cascading impacts on oy-
1961). It has been well documented that small crustaceans are a key ster reefs, diminishing their economic and ecological value. In order to
link in the transfer of energy between primary producers and secondary better understand these processes, we must place future efforts on the
consumers and that these species compose a large proportion of the effects of combined stressors, including pesticides, on oyster reef
diets of juvenile fish and other mid-level predators (Adams, 1976; Edgar community composition and overall ecosystem function.
and Aoki, 1993; Yeager and Layman, 2011; Abeels et al., 2012). A
stable isotope study by Abeels et al. (2012) revealed that crustaceans CRediT authorship contribution statement
are included in all lower trophic levels and were consumed by every
secondary consumer studied on a sub-tropical oyster reef. An in- Samantha M. Iliff: Conceptualization, Data curation, Formal ana-
troduction of pesticides into an estuarine ecosystem could potentially lysis, Investigation, Methodology, Project administration, Resources,
impact the abundance and composition of crustacean communities. A Software, Supervision, Visualization, Writing - original draft, Writing -
shift at this trophic level could result in cascading effects throughout review & editing. Rachel J. Harris: Conceptualization, Methodology,
the ecosystem including a reduction in biomass, a loss of diversity, and Project administration, Resources, Supervision, Writing - review &
a decrease in available prey for fish and other secondary consumers. editing. Elizabeth W. Stoner: Formal analysis, Investigation,
Although there were no significant differences in species diversity Methodology, Project administration, Resources, Software, Supervision,
and richness between the treatments, there are several mechanisms that Visualization, Writing - review & editing.
could potentially explain these variable results. Based on the average
salinity (11.2 ppt) and the dissolved fraction of the dissolution blocks, Acknowledgements
we estimate an average flow velocity of 15 cm/s over the course of this
study (Thompson and Glenn, 1994), which falls within the average The authors wish to thank the Loxahatchee River Environmental
range 5–25 cm/s of flow velocities reported on oyster reefs (Reidenbach Control District, United States for funding. We also wish to thank
et al., 2013), and within the maximum current velocities (17 cm/s) Katherine Banger, Owen Silvera, Aaron Kent, Kyle Vincent, Liberty
previously measured in the South West fork of the Loxahatchee River Boyd, Susan Noel, Helen Johnson, David Porter, and Jerry Metz for field
Estuary (Patra, 2003). Although low compared to high energy systems, and data assistance; Eric Lazo-Wasem for his assistance identifying
over time, these flow rates would have caused the carbaryl to dissipate epifauna; Jon Moore and Jim Wetterer for edits to the original under-
rapidly; averaging 2.6 ( ± 0.3) g carbaryl/day. Thus, we regularly re- graduate thesis manuscript; and an anonymous review for their com-
placed the blocks (every 3–5 days) to achieve chronic exposure over the ments and suggestionson.
28-day study period. We did not measure carbaryl in ambient waters
during this study. However, with a solubility of 110,000 μg/L in es- Funding
tuarine waters (Poore et al., 2009), and a dissolution rate of 2.6 g/day,
we estimate that the average concentrations of carbaryl were approxi- This work was supported by the Loxahatchee River District.
mately 8800 μg carbaryl/L; although the concentrations would have
varied greatly throughout the tidal cycle (0–110,000 μg/L). On average Declaration of Competing Interest
the carbaryl concentrations reported here are much higher than 100 μg/
L reported in well-mixed estuarine waters (Schroeder-Spain, 2017), and None.
even on the high end of acute (24–48 h) exposure limits reported for
invertebrate species (30–7200 μg carbaryl/L; Feldman et al., 2000). References
These mean concentrations of carbaryl (8800 μg/L) seem quite high
compared to the acute toxicity studies, yet, unlike the acute toxicity Abeels, H.A., Loh, A.N., Volety, A.K., 2012. Trophic transfer and habitat use of oyster
studies, this experiment was not conducted in a closed system. Thus, Crassostrea virginica reefs in southwest Florida, identified by stable isotope analysis.
Mar. Ecol. Prog. Ser. 462, 125–142.
despite regularly replacing the carbaryl dissolution blocks it is likely Adams, S.M., 1976. Feeding ecology of eelgrass fish communities. Trans. Am. Fish. Soc.
that the variations in flow direction and velocities dissipated the ef- 105 (4), 514–519.
fectiveness of our carbaryl treatments (Whalen et al., 2013). Therefore, Anderson, M.J., 2001. A new method for non-parametric multivariate analysis of var-
iance. Austral. Ecol. 26 (1), 32–46.
it is possible that some applications of carbaryl could enter the oyster Anderson, B.S., Hunt, J.W., Phillips, B.M., Tjeerdema, R.S., 2007. Navigating the TMDL
reef at higher concentrations than those used here. In turn, this would Process: Sediment Toxicity. 02-WSM-2. Water Environment Research Foundation, pp.
magnify impacts to the oyster reef community. 194.
Anderson, B., Phillips, B., Hunt, J., Siegler, K., Voorhees, J., Smalling, K., Kuivila, K.,
Oyster reefs are one of the most ecologically and economically va- Hamilton, M., Ranasinghe, A., Tjeerdema, R., 2014. Impacts of pesticides in a Central
luable habitats within estuaries. It is not well known why oyster reefs California estuary. Environ. Monit. Assess. 186 (3), 1801–1814.
are in decline, yet it is apparent that a combination of stressors could be Armhurst, K.L., Grosby, D.G., 1991. Fate of carbaryl, 1-naphthol and atrazine in seawater.
Pac. Sci. 45, 314–320.
in effect. Here, we were able to detect the negative effects of chronic
Bahr Jr., L.M., 1976. Energetic aspects of the intertidal oyster reef community at Sapelo
pesticide exposure on crustaceans. Additional studies must focus on Island, Georgia (USA). Ecology 57 (1), 121–131.
acute versus chronic effects of pesticides on estuarine communities. Our Bahr, L.M., Lanier, W.P., 1981. The Ecology of Intertidal Oyster Reefs of the South
exposure time was limited to 28 days and it is possible that carbaryl Atlantic Coast: A Community Profile (No. 81/15). US Fish and Wildlife Service.
Barnes, T.K., Volety, A.K., Chartier, K., Mazzotti, F.J., Pearlstine, L., 2007. A habitat
could have greater impacts on estuarine community composition over a suitability index model for the eastern oyster (Crassostrea virginica), a tool for re-
larger time period. Furthermore, carbaryl could be a relatively mild storation of the Caloosahatchee Estuary, Florida. J. Shellfish Res. 26 (4), 949–960.
pesticide in marine environments with variable toxicity and a relatively Bay, S.M., Mikel, T.M., Schiff, K., Mathison, S., Hester, B., Young, D., Greenstein, D.,
2005. Southern California Bight Regional Monitoring Program: i. Sediment Toxicity.
short half-life (Armhurst and Grosby, 1991). Future work must consider Technical Report 451. Southern California Coastal Water Research Project,
the response of both individual species and the entire community to Westminster.
various degrees of pesticides to determine specific toxicities and toler- Beck, M.W., Brumbaugh, R.D., Airoldi, L., Carranza, A., Coen, L.D., Crawford, C., Defeo,
O., Edgar, G.J., Hancock, B., Kay, M.C., Lenihan, H.S., 2011. Oyster reefs at risk and
ances. Additionally, mixtures of pesticides commonly used in agri- recommendations for conservation, restoration, and management. Bioscience 61 (2),
cultural and gardening practices including pyrethroids and organo- 107–116.
phosphates could increase the toxicity to marine species, lingering in Canty, M.N., Hagger, J.A., Moore, R.T.B., Cooper, L., Galloway, T.S., 2007. Sublethal
impact of short term exposure to the organophosphate pesticide azamethiphos in the
the water and sediment, or bioaccumulating through trophic levels. marine mollusc Mytilus edulis. Mar. Pollut. Bull. 54 (4), 396–402.
Future studies should incorporate other pesticides such as herbicides Cook, K., Vanderklift, M.A., Poore, A.G., 2011. Strong effects of herbivorous amphipods
and algaecides to consider the impacts on sensitive marine habitats. on epiphyte biomass in a temperate seagrass meadow. Mar. Ecol. Prog. Ser. 442,
507
S.M. Iliff, et al. Marine Pollution Bulletin 146 (2019) 502–508
508