Journal of Experimental Marine Biology and Ecology 440 (2013) 3541

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Journal of Experimental Marine Biology and Ecology

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Effect of a physical disturbance event on deep-sea nematode community structure and ecosystem function
Daniel Leduc a, b,, Conrad A. Pilditch c
a b c

Department of Marine Science, University of Otago, P.O. Box 56, Dunedin, New Zealand National Institute of Water and Atmospheric Research, Private Bag 14-901, Wellington 6021, New Zealand Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton, New Zealand

a r t i c l e

i n f o

a b s t r a c t
Numerous studies have been conducted on the effect of physical disturbance on shallow water benthic communities, but there is a paucity of data from deep-sea environments. We conducted a laboratory experiment using undisturbed sediment cores from Chatham Rise (water depth=345 m), Southwest Pacic, to investigate the effects of a physical disturbance event (resuspension of surface sediments) on sediment characteristics (sediment grain size, pigment content), nematode community attributes (abundance, diversity, community structure) and ecosystem function (sediment community oxygen consumption (SCOC)) over a period of 9 days. Disturbance did not have any noticeable impact on sediment characteristics, SCOC, or nematode species richness, but led to changes in vertical distribution patterns and shifts in nematode community structure. The magnitude of disturbance-related effects was, however, much smaller than the effect of sediment depth (01, 13, and 35 cm), and the main impact of disturbance on nematode vertical distribution patterns and community structure appeared to be related to a vertical re-shufing of nematodes in the sediments rather than mortality. We did not observe substantial increases in the abundance of nematode genera generally regarded as disturbance-tolerant, such as Sabatieria. The worst-affected species belongs to the Stilbonematinae, a group of typically long and slender nematodes that may be easily damaged by physical disturbance. The limited impact of physical disturbance on benthic community structure and function suggests that the Chatham Rise nematode community is relatively resilient to sediment resuspension. This resilience may have arisen from frequent exposure to disturbance in the eld (e.g., from strong currents), or may be a more widespread feature of nematode communities. 2012 Elsevier B.V. All rights reserved.

Article history: Received 19 September 2012 Received in revised form 20 November 2012 Accepted 20 November 2012 Available online xxxx Keywords: Biodiversity Chatham Rise Continental slope Laboratory experiment Meiofauna Resilience

1. Introduction Disturbance has long been recognised as an important factor structuring communities (Connel, 1978; Petraitis et al., 1989). Numerous studies have shown that disturbance can inuence the abundance, diversity, and community structure of faunal (e.g., Austen et al., 1998; Schratzberger et al., 2009) and microbial communities (e.g., Cowie and Hannah, 2007; Findlay et al., 1990) in marine benthic environments. Changes in the structure of benthic communities are, in turn, likely to inuence ecosystem processes such as secondary productivity and nutrient cycling (Danovaro et al., 2008; Karlson et al., 2010; Pusceddu et al., 2005). The relationship between disturbance, benthic community structure, and ecosystem function, however, is likely to be complex, and our knowledge of the processes involved remains patchy (Lee et al., 2001; Snelgrove, 1997).

Corresponding author at: National Institute of Water and Atmospheric Research, Private Bag 14-901, Wellington 6021, New Zealand. Tel.: +64 4 386 0300x7074. E-mail address: Daniel.Leduc@niwa.co.nz (D. Leduc). 0022-0981/$ see front matter 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.jembe.2012.11.015

Physical disturbance, dened as the disruption of sediment particles caused by abiotic (e.g., currents), biotic (e.g., bioturbation), and anthropogenic processes (e.g., bottom-trawling), is widespread in benthic systems and occurs at a variety of spatial and temporal scales (Hall, 1994; Kaiser, 1998; Watling and Norse, 1998). Frequent and/or widespread disturbance, such as that caused by bottom trawling, has been linked to markedly lower standing stocks and diversity of coastal benthic communities (Hinz et al., 2008; Reiss et al., 2009), as well as pronounced shifts in function of soft sediment habitats (Pilskain et al., 1998; Puig et al., 2012; Reiss et al., 2009; Thrush and Dayton, 2002; Watling and Norse, 1998). At the local scale, disturbance may alter sediment grain size structure, biogeochemical gradients, and availability of food resources, thereby favouring some species and inhibiting others (Thistle, 1981; Widdicombe and Austen, 2001). Relatively little is known, however, about the short-term response of deep-sea infaunal communities to local physical disturbance (Anhert and Schriever, 2001). We could expect, given the stable nature of deep-sea ecosystems relative to their shallow counterparts, that a high proportion of species will be negatively affected by disturbance, resulting in a short-term decline in abundance and species richness (Grassle and Sanders, 1973). Any

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negative impact on species richness, could, in turn, inuence ecosystem functioning (Danovaro et al., 2008). Most studies on the effect of physical disturbance on marine benthic communities have been conducted in relatively shallow waters (Hall, 1994; Thrush and Dayton, 2002). Quantifying the effect of disturbance on deep-sea benthic communities, however, remains a challenging task due to logistical reasons. The relatively low macroand megafaunal densities in deeper waters require obtaining a greater number of samples of relatively large volume in order to detect ecologically meaningful change (Gage and Bett, 2005; Rogers et al., 2008; Schratzberger et al., 2000), and conducting experiments in deep-sea settings is difcult and costly. Field experiments on the impact of deep-sea mining on the benthos have yielded some insights into the response of deep-sea communities to physical disturbance (e.g., Anhert and Schriever, 2001; Miljutin et al., 2011; Thiel and Schriever, 1990), but few laboratory experiments have been conducted on the response of deep-sea organisms/communities to physical disturbance (Gallucci et al., 2008a). Meiofaunal organisms, and nematodes in particular, are well suited for laboratory experiments because of their small size, high abundance and wide distribution (Giere, 2009). Experiments can be conducted on deep-sea nematode communities comprising hundreds to thousands of individuals using relatively small volumes (b 1 L) of sediment (Gallucci et al., 2008a). This approach is less costly than conducting experiments in situ and provides greater control over experimental condition. Laboratory experiments are among the best tools for improving our understanding of the short-term impacts of disturbance on community dynamics and ecosystem function at the scale of the disturbed patch, and can provide new insights into the mechanisms involved (Schratzberger and Warwick, 1999; Schratzberger et al., 2009). Such experiments could, in addition, help identify which taxa are more susceptible to disturbance (e.g. Schratzberger and Warwick, 1999), thereby underpinning the use of nematodes for monitoring of deep-sea benthic ecosystems (Miljutin et al., 2011). The aim of the present study is determine the short-term (b 10 days) effect of a one-off physical disturbance event on the abundance, species richness, and community structure of a bathyal nematode community using a laboratory experiment. More specically, we test the hypothesis that physical disturbance leads to mortality in a majority of potentially disturbance-sensitive nematode taxa, resulting in lower abundance, species richness, and altered community structure. The effect of disturbance on ecosystem function was also investigated by comparing sediment community oxygen consumption (SCOC) between disturbed and undisturbed treatments. 2. Methods 2.1. Study area and sampling The study site (43 20.1 S, 178 17.5 E) was located at 345 m water depth on the crest of Chatham Rise, a submarine ridge extending eastwards from the South Island of New Zealand (Fig. 1). The rise encompasses water depths from ca. 250 to 3000 m and lies beneath the Subtropical Front (STF), a region associated with heightened primary productivity (Bradford-Grieve et al., 1997; Murphy et al., 2001) and high levels of mixing and current activity (Nodder et al., 2007; Sutton, 2001). Sediments on the crest of the rise are characterised by deposits of authigenic and biogenic silty sands with localised accumulations of phosphorite nodules (Cullen, 1987; Orpin et al., 2008), which are being targeted by extractive industry. Samples were collected on February 20, 2011 during National Institute of Water and Atmospheric Research (NIWA) cruise TAN1103 using an Ocean Instruments MC-800A multicorer (MUC; core internal diameter = 9.5 cm). Faunal samples were obtained from three cores (from 2 MUC deployments) and processed on board ship as described below to provide eld controls. An additional twelve cores from eight MUC

Fig. 1. Map of study area (east of New Zealand's South Island) showing position of study site (circle) and location of Subtropical Front (dotted lines).

deployments were obtained for the disturbance experiment. The upper 811 cm of sediment and the overlying water from each core were transferred into incubation units (internal diameter = 8.4 cm, height= 30.0 cm) by subcoring (Nodder et al., 2003). Incubation units were then sealed, kept in the dark at bottom water temperature (8.0 C0.5 C,), and aerated to ensure adequate oxygen supply. Incubation units were transferred to a water bath (8.0 C 0.5 C) in the laboratory 5 days after sampling, and were connected to a recirculating seawater system at same temperature for two additional days prior to the beginning of the experiment. Constant seawater ow ensured adequate oxygenation of the incubation units (D. Leduc, unpublished data) but did not disturb the sediment surface. 2.2. Disturbance experiment Six incubation units were randomly allocated to an undisturbed treatment (operational controls) and six were allocated to a disturbed treatment. Three incubation units from each treatment were destructively sampled at 2 and 9 days after disturbance. Physical disturbance of the sediments was achieved by re-suspending the upper 5 cm of sediments with a plastic paddle for 5 seconds. Sediments were left to settle for 8 hours before reconnecting incubation units to recirculating seawater system. Samples for estimation of nematode community attributes (i.e., abundance, species richness, and community structure) were obtained from disturbed and undisturbed treatments as well as from eld controls. Each sample consisted of one subcore of internal diameter 29 mm taken to a depth of 5 cm. Subcores were sliced into 01, 13, and 35 cm depth layers and xed in 10% formalin and stained with Rose Bengal. Samples were subsequently rinsed on a 1 mm sieve to remove large particles and on a 45 m sieve to retain nematodes. Nematodes were extracted from the remaining sediments by Ludox otation and transferred to pure glycerol (Somereld and Warwick, 1996). Between 110 and 150 randomly chosen nematodes (or all individuals if fewer were present in the sample) were mounted on permanent slides and identied to genus and putative species using the descriptions in Warwick et al. (1998), as well as the primary literature. Monhystrella and Thalassomonhystera were treated as one genus (Monhysteridae) because they are sometimes difcult to distinguish based on morphology (Fonseca and Decraemer, 2008). The remaining fauna in the sample were mounted in glycerol on a separate slide and the abundance of nematodes was quantied using a compound microscope (100 magnication). Nematode abundance was compared between treatments and sediment depths as nematode density (ind. cm 3) to take into account the different volumes of the sediment depth layers. Nematode species richness was quantied using Hurlbert's expected number of species for a sample of 51 individuals (ES(51), Hurlbert, 1971).

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Samples for the analysis of sediment parameters were obtained from disturbed and undisturbed treatments using a small subcore (19 mm internal diameter) to a depth of 5 cm. Subcores were sliced into 01, 13, and 35 cm depth layers and kept frozen at 20 C until analysis. The following sediment parameters were determined: chloroplastic pigment equivalents (CPE; sum of chlorophyll a and phaeopigments), geometric mean grain size, particle sorting, and % sand. Methods for the analyses of sediment parameters are given in Nodder et al. (2003) and Grove et al. (2006). Sediment community oxygen consumption (SCOC) was measured immediately prior to disturbance (t = 0), 2 days after disturbance (t = 2), and 9 days after disturbance (t = 9). Water ow was stopped and the incubation units were sealed prior to SCOC determination. Incubations were conducted in the dark for a period of 24 hours, at which point oxygen levels had decreased by less than 10% of initial levels. A magnetically driven impellor (~ 60 rpm) tted to the chamber lids gently circulated water during the incubation. Oxygen concentration was measured with a pre-calibrated PreSens MICROX I microoptode inserted through a sampling port in the chamber lid. Three more O2 measurements were made during the incubation period. SCOC was estimated from the decline in O2 concentration with time (linear regression R 2 > 0.9). 2.3. Data analyses The PERMANOVA routine in PRIMER was used for the analysis of univariate and multivariate data (Anderson et al., 2008). PERMANOVA is a semi-parametric, permutation-based routine for analysis of variance based on any similarity measure (e.g., Euclidean, BrayCurtis). Because the sediments in four of our incubation units were obtained from the same MUC deployment (i.e., they are pseudoreplicates), we tested for the potential effect of MUC deployment on nematode community attributes prior to the main analyses. No statistically signicant differences were found (BrayCurtis similarity measure of square root-transformed data, P >0.1). For the analysis of nematode community attributes, we began by comparing variables between eld and operational controls (day 2 and 9) to evaluate the impact of laboratory conditions on the nematode community. These analyses were conducted using a repeated measure design (to take into account the lack of independence between sediment depths) using the xed factor Control (three levels: eld control and operational controls at day 2 and 9), the xed factor Depth (three levels: 01, 13, and 35 cm), and with replicates nested within Control but not Depth (Quinn and Keough, 2009). The effect of disturbance on sediment parameters, nematode community attributes, and SCOC was investigated by comparing variables in the disturbed and undisturbed treatments 2 and 9 days after disturbance whilst omitting eld controls from the analyses. These analyses were conducted using the xed factor Disturbance (two levels: disturbed and undisturbed), the xed factor Time (two levels: day 2 and 9), the xed factor Depth (three levels: 01, 13, and 35 cm), and with replicates nested within Disturbance and Time but not Depth (repeated measure design, Quinn and Keough, 2009). Because nematode community attributes are often correlated with environmental factors such as food availability and sediment granulometry (e.g., Leduc et al., 2012a), environmental parameters (CPE, mean grain size, sorting, and %sand) were tted rst as covariates, followed by the factors listed above. Sequential sum of squares was used to take into account the sequential order of terms (i.e., covariates rst, followed by the factors; Anderson et al., 2008). Similarity matrices for univariate variables (i.e., nematode density and species richness, sediment parameters, SCOC) were built using Euclidean distance of untransformed data, and similarity matrices for multivariate data (nematode community structure) were built using the BrayCurtis similarity measure of square root-transformed data (Anderson et al., 2008). P-values for individual predictor variables were obtained using 9999 permutations, and

P-values for pairwise comparisons were obtained using Monte-Carlo sampling due to the low number of possible permutations (Anderson et al., 2008). The square root of estimates of components of variation was used for comparing the amount of variation attributable to different terms in the multivariate PERMANOVA models (Anderson et al., 2008). The SIMPER routine in PRIMER was used to identify the species contributing most to within-group similarity and between-group dissimilarity (Clarke and Warwick, 2001). 3. Results A total of 4412 nematodes belonging to 362 species and 146 genera were identied. Eighty one (22%) of these species were recorded only once, and 54 (15%) were recorded twice. The genera with highest overall mean abundance were: Microlaimus (41 ind. 10 cm 2), Sabatieria (35), Molgolaimus (27), Daptonema (20), Monhysteridae (19), Halalaimus (19), and Desmoscolex (19). No obvious vertical gradient in sediment colouration was observed in any of the experimental units throughout the experiment, which suggests that the RPD layer depth exceeded the height of the sediments in the experimental units (8.010.5 cm). Live macrofauna, including polychaetes, amphipods, and gastropods, were present in the experimental units throughout the experiment. Some mortality, however, appears to have occurred, as suggested by the development of small black patches of anoxic sediments below the surface of most cores by the end of the experiment. No obvious differences in the number or size of these patches were observed between disturbed and undisturbed experimental units. 3.1. Effect of experimental conditions Total, depth-integrated (05 cm) nematode abundance in the eld and operational controls ranged from 1311 to 2902 ind. 10 cm 2 (mean= 1969 ind. 10 cm 2). Total abundance was signicantly lower at the end of the experiment relative to 2 days after disturbance and eld controls (1492 vs. 1937 and 2494 ind. 10 cm 2, respectively, one-way PERMANOVA and pairwise comparisons, P b 0.05). Nematode density (ind. cm 3) differed signicantly between sediment depths (two-way PERMANOVA, P b 0.05, Table 1) but not between controls. Pairwise comparisons showed that this difference between sediment depths was due to higher nematode density in the 01 cm relative to the 35 cm layer (mean 108 vs. 11 ind. cm 3 respectively, P b 0.05). Nematode density in the 13 cm layer was intermediate and did not differ signicantly from the other two layers (33 ind. cm 3, P > 0.05). Nematode species richness (ES(51)) did not differ signicantly between controls, but differed signicantly between sediment depths (P b 0.05, Table 1). Pairwise comparisons showed that this difference was due to signicantly lower species richness in the 35 cm layers relative to the 01 and 13 cm layers (ES(51): 27.8 vs. 33.4 33.7). Nematode community structure differed signicantly between controls, sediment depths, and their interaction (Table 1). The presence of a signicant interaction suggests that differences in community structure among controls were not consistent among sediment depths (Fig. 2); pairwise comparisons, however, did not reveal any signicant differences between any combinations of controls and sediment depth (P > 0.05, data not shown). Differences in community structure between controls were the result of relatively minor shifts in the abundance of species (SIMPER, Appendix 1). Comparison of the square-root of estimates of components of variation indicated that the effect of sediment depth on nematode community structure was 3.0 and 1.9 times stronger than the effects of control type and interaction term, respectively. Values of SCOC ranged from 170.2 to 364.7 mol m 2 h 1 (mean= 266.1 mol m 2 h 1) and did not vary signicantly between operational controls at 2 and 9 days (data not shown, P > 0.05).

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D. Leduc, C.A. Pilditch / Journal of Experimental Marine Biology and Ecology 440 (2013) 3541 Table 2 Sediment parameters (mean (standard deviation)) at 01, 13, and 35 cm sediment depth in the incubation units (N = 12). Values followed by different letters are signicantly different between sediment depths. See Appendix 1 for results of PERMANOVA. CPE: chloroplastic pigment equivalents. CPE (ng cm 3) 01 cm 13 cm 35 cm 6.14 (1.44) 4.60 (1.33)b 4.61 (1.72)b
a

Table 1 Results of PERMANOVA analyses testing for the effect of control (eld controls and operational controls at day 2 and 9), sediment depth (01, 13, and 35 cm) and their interaction on nematode community attributes. Variables that are signicantly correlated with the response variable are shown in bold. Source Density Control Depth Replicate (Control) Control Depth Residuals Total Control Depth Replicate (Control) Control Depth Residuals Total Control Depth Replicate (Control) Control Depth Residuals Total df 2 2 6 4 12 26 2 2 6 4 12 26 2 2 6 4 12 26 MS 1113 23242 264 168 356 3 101 8 15 17 3294 10249 2340 2568 1748 Pseudo-F 4.22 65.21 0.74 0.47 P 0.093 0.001 0.677 0.754

Mean grain size (m) 43.9 (16.3) 59.4 (22.2)b 54.7 (14.1)b
a

Sorting 4.01 (0.27) 4.09 (0.51)a 4.29 (0.47)a

a

%Sand 45.9 (13.4)a 56.6 (13.1)b 55.1 (9.0)b

Species richness (ES(51))

0.41 5.88 0.46 0.88

0.702 0.026 0.818 0.539

Community structure

1.41 5.86 1.34 1.50

0.020 0.001 0.012 0.009

3.2. Effect of disturbance There was no signicant effect of disturbance or sampling time (2 vs. 9 days after disturbance) on sediment parameters (Appendix 2). There was, however, a signicant effect of sediment depth on CPE, mean grain size, and % sand (PERMANOVA, P b 0.05). Pairwise comparisons showed that CPE values were greater in the surface (01 cm) layer relative to subsurface (13 and 35 cm) layers, whilst mean grain size and %sand were lower in the surface layer relative to subsurface layers (P b 0.05, Table 2). There were no differences in SCOC between disturbance treatments and controls, or between sampling times (data not shown, P > 0.2).

Nematode density (ind. cm 3) was signicantly correlated with the covariates CPE and mean grain size (PERMANOVA, P b 0.01, Appendix 3). There was a signicant effect of disturbance, sediment depth, and their interaction on nematode density (P b 0.05). Nematode density in surface (01 cm) sediments was signicantly lower in disturbed experimental units relative to undisturbed units, whilst the opposite trend was observed in the deepest (35 cm) sediment layer (pairwise comparisons, P b 0.05, Fig. 3). Total, depth-integrated abundance (ind. 10 cm 2), however, did not differ between the disturbed treatment and the controls (data not shown, P >0.05). There was a signicant effect of sediment depth on species richness, with values in the 35 cm layer signicantly lower than in the 01 and 13 cm layers (P b 0.05, Table 3, Appendix 3), but disturbance did not have a signicant effect (P > 0.05). Nematode community structure was signicantly correlated with the covariate CPE, but not with sediment grain size parameters (Appendix 3). There were signicant differences in community structure between disturbed treatments and controls, between sediment depths, but not sampling times (P b 0.05, Fig. 4). Comparison of the square-root of estimates of components of variation for disturbance (11.3) and sediment depth (27.8) indicated that sediment depth accounted for two and a half times more of the variation in nematode community structure than disturbance. About a third of the within-group similarity of surface sediments (01 cm) was accounted for by species of the family Desmoscolecidae (Desmolorenzenia sp. 1, Desmoscolex sp. 1, and Hapalomus sp. 1), Calomicrolaimus sp. 1, and Chromadorita sp. 1 (Appendix 4). Molgolaimus sp. 1, Nannolaimoides sp. 1, and Stilbonematinae sp. 2 accounted for much of the within-group similarity of the 13 cm sediment depth layer, whilst Stilbonematinae sp. 2, Neotonchus sp. 2, and Laimella sp. 1 were the highest contributors to within-group similarity of the 35 cm sediment depth layer. Species contributing most to within-group similarity also explained much of the dissimilarity between sediment depth layers (Appendix 5). Nannolaimoides sp. 1, Stilbonematinae sp. 2, Southernia sp. 1 explained much of the within-group similarity for both the disturbed and undisturbed treatments. Disturbed sediments, however, were characterised by lower abundance of Stilbonematinae sp. 2, Laimella sp. 1,, and Southernia sp. 1 than undisturbed sediments, and higher abundance of Neotonchus sp. 2, Nannolaimoides sp. 1, Desmoscolex sp. 1, and Calomicrolaimus sp. 1 (Appendix 4 and 5). 4. Discussion 4.1. Experimental conditions The experimental units did not allow for immigration of nematodes from nearby sediments or from the water column, even though these processes are likely to be important in natural settings (e.g., Le Guellec, 1988; Sun and Fleeger, 1994). The aim of our experiment, however, was to investigate the short-term impact of disturbance on local community dynamics at the scale of the disturbed patch. The severity of the articial disturbance could be compared to bioturbation induced by benthic megafauna or bottom feeding sh, or a resuspension event caused by strong currents. The articial conditions created within the experimental units did not affect nematode species richness or density (or their vertical

Fig. 2. Two-dimensional multidimensional scaling conguration for nematode species abundance (square-root transformed data) showing differences between (A) sediment depths (01, 13 and 35 cm), and (B) eld controls and operational controls 2 and 9 days after the beginning of the experiment.

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Fig. 3. Mean nematode density (ind. cm 3) in disturbed and undisturbed treatments at different sediment depths (01, 13, and 35 cm) and at different sampling times (t = 2 and 9 days after beginning of experiment). Error bars are standard deviations from the mean.

gradients), but led to lower total nematode abundance and shifts in community structure at the end of the experiment. The changes in nematode community attributes we observed in the disturbed treatment were, therefore, an integrated response to both the conditions inside the experimental units and the physical disturbance itself. Declines in nematode abundance inside microcosms relative to the eld have been observed in shallow water-based laboratory experiments, and are usually associated with marked declines of specic taxa (e.g., chromadorids; Schratzberger and Warwick, 1999; Schratzberger et al., 2000). The changes in community structure we observed, however, were relatively complex (as suggested by the presence of a signicant interaction between sediment depth and control type), and resulted from small changes in species relative abundances rather than changes in species composition (see Appendix 1). Reasons for the decline in nematode abundance over the experimental duration remain unclear, but are unlikely to be related to changes in food availability since we did not observe any changes in pigment concentrations over the course of the experiment. Predation by macrofauna may have played a role, but this process was not quantied in the present study. 4.2. Effect of disturbance Disturbance did not inuence pigment content and sediment granulometry in the experimental units, or their vertical distribution. This nding was unexpected given the relatively severe nature of the disturbance which led to complete resuspension of the top 5 cm of sediments. These results could indicate that the sediments at the Chatham Rise study site are subjected to regular disturbance events. Periodic
Table 3 Nematode species richness (mean (standard deviation)) in disturbed and undisturbed treatments, and at 01, 13, and 35 cm sediment depths in the incubation units (averaged across sampling times). Values followed by different letters/number are signicantly different between treatment/sediment depths. See Appendix 3 for results of PERMANOVA. ES(51) Treatment Undisturbed Disturbed Sediment depth 01 cm 13 cm 35 cm 31.6 (5.3)a 34.2 (3.9)a

Fig. 4. Two-dimensional multidimensional scaling conguration for nematode species abundance (square-root transformed data) showing differences between (A) sediment depths (01, 13 and 35 cm), and (B) disturbed and undisturbed treatments.

34.6 (3.2)1 33.5 (4.6)1 30.7 (5.6)2

resuspension of sediment particles would explain the presence of a surface layer with greater content of lighter ne particles than subsurface layers, as we observed in the experimental units at the beginning of the experiment. The presence of relatively high pigment concentrations in the deepest (35 cm) sediment layer is also suggestive of physically-mixed sediments (see Table 2; Sun and Dai, 2005). Near-bottom current speeds on and near Chatham Rise crest regularly exceed erosion thresholds (Nodder et al., 2007), and lead to periodic resuspension of surface sediments. Bioturbation by macrofauna could also help explain the relatively high pigment concentrations in subsurface sediments (Sun and Dai, 2005). Lack of impact of disturbance on sediment community oxygen consumption (SCOC) provides additional support to the notion of a naturally disturbed community, although this observation may be better explained by the relatively coarse nature of the sediments with a deep RPD layer (>8 cm below sediment surface). Coarse sediments are typically well-oxygenated with weak vertical biogeochemical gradients and are therefore not as severely impacted by physical disturbance as ner sediment communities (Glud, 2008). Similar results were obtained in microcosm study by Braeckman et al. (2011) on a subtidal ne sand community, which showed no or only limited impact of a physical disturbance event on SCOC and pigment concentration distribution. Disturbance did not inuence nematode survival, as indicated by the absence of disturbance effect on total depth-integrated nematode abundance. The presence of signicant disturbance, sediment depth, and interaction effects on nematode densities, however, suggests that disturbance led to a vertical re-shufing of nematodes. More specically, physical disturbance appears to have led to the burial of nematodes from the surface sediment layer into subsurface layers (see Fig. 3). A similar response was observed for a physically disturbed subtidal nematode community (Braeckman et al., 2011). Experimental evidence suggests limited impact of physical disturbance on nematode survival (Schratzberger et al., 2002), and that, when present, impacts are most pronounced in communities inhabiting muddy sediments than sandy sediments (Schratzberger

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and Warwick, 1998). Our ndings, based on an upper slope muddy sand community from Chatham Rise, are consistent with the latter studies. Our results, however, may partly reect the low level of replication (N = 3) and high variability in nematode abundance between experimental units, which limit the power of the present study. Experiments on the effects of disturbance on nematode communities, in comparison, typically use homogenised and/or sieved sediments, which reduces variability between experimental units (e.g., Braeckman et al., 2011; Schratzberger and Warwick, 1999). This kind of manipulation, however, constitutes disturbance in itself, and may impact nematode communities before the beginning of the experiment. Clear shifts in nematode species richness were observed between sediment layers, with lower values in the deepest (35 cm) layer relative to the two uppermost layers (01 and 13 cm). This is a common, though not universal, trend observed in shallow water and deep-sea sediments (e.g., Ingels et al., 2011; Leduc and Probert, 2011), and may be related to the more challenging environmental conditions often found in deeper sediment layers (e.g., low oxygen levels, high sulphide concentrations; Vanaverbeke et al., 2011). Larger, more mobile taxa that do not feed selectively may also be better at exploiting the often limited and low quality food sources in deeper sediments (Ingels et al., 2011; Soetaert et al., 2002). We observed greater species richness in disturbed sediments relative to undisturbed sediments; this trend is most likely due to the vertical mixing of nematodes between sediment layers (see above), which would lead to vertical homogenisation of species distributions, thereby increasing the pool of species in a given layer (e.g., Leduc et al., 2010). The evidence available to date suggests that the effect of disturbance on local nematode diversity depend on the interaction of factors such as type/frequency of disturbance (Austen et al., 1998; Schratzberger and Warwick, 1998) and sediment grain size (Schratzberger and Warwick, 1998). The limited impact of disturbance on nematode species richness in the present study may be related to the low frequency of disturbance (only once) and the relatively coarse sediments at the Chatham Rise study site. We found a signicant effect of disturbance on nematode community structure, although the magnitude of this effect was small relative to the effect of sediment depth. This effect could be partly explained by the vertical mixing of individuals in the experimental units. The effect of disturbance was relatively subtle, and was due to small shifts (in both directions) in the abundance of many species. Some genera, such as Sabatieria and Leptolaimus, are known to respond positively to disturbance in shallow water systems, while others, such as Desmodora, are often negatively affected (see meta-analysis by Schratzberger et al., 2009). These genera were present in the experimental units but we did not observe substantial changes in their relative abundances. Of all the species present in our experimental units, Stilbonematinae sp. 2 was the most adversely affected by the disturbance event. This species, as well as all other species within the subfamily Stilbonematinae, have long, slender bodies that may be easily damaged by physical disturbance. Species of this group are typically covered by a dense layer of bacterial sulphur-oxidising ectosymbionts and are seldom reported from deep-sea habitats (but see Tchesunov et al., 2012; Van Gaever et al., 2006). Species of Stilbonematinae are usually found in shallow water coarse sediments with high organic matter input, and concentrate at the interface between oxygenated surface sediments and deeper, suboxic sediments (Ott and Novak, 1989). Their presence in relatively high densities, particularly in the deepest sediment layer (where they represent up to 17% of nematode abundance), may suggest the presence of reduced subsurface conditions, even though we did not observe a RPD layer in the experimental units. Stilbonematinae sp. 2 has been recorded from several other sites along Chatham Rise crest (D. Leduc, unpublished data), a distribution pattern probably related to the proximity of the highly productive Subtropical Front (Bradford-Grieve et al., 1997; Grove et al., 2006). Further

research is required to ascertain the impact of physical disturbance on this species. The Chatham Rise nematode community appears to be resilient to disturbance. This resilience may be due to frequent exposure to disturbance in the eld, or may be a common feature of nematode communities in general. As discussed above, lack of change in sediment parameters and SCOC following disturbance is consistent with periodically disturbed sediments. Schratzberger and Warwick (1999, p.227) noted that (nematode) assemblages are most affected by the kinds of disturbances that they do not normally experience naturally, which also supports the notion of a naturally disturbed community. The nematode community at the study site, however, was characterised by relatively high diversity (e.g., Leduc et al., 2012b), whilst disturbed communities typically have low diversity with high dominance of a few resilient taxa such as Sabatieria (Schratzberger et al., 2009). This genus was neither dominant nor particularly abundant in our samples. It is possible that the disturbance was not sufciently severe (in intensity or frequency) to elicit a response. Several laboratory experiments, for example, were designed to investigate the effect of periodic physical disturbance on nematode communities (e.g., Schratzberger and Warwick, 1998, 1999), as opposed to a one-off physical disturbance event in the present study. Studies in shallow water environments have shown that nematodes are more resilient to disturbance than larger macrofaunal organisms (Schratzberger et al., 2002; Whomersley et al., 2009), but more research is needed to determine how resilient deep-sea nematode communities may be to physical disturbance relative to shallow water communities. The relatively mild physical disturbance caused by bioturbation or resuspension events does not appear to have a major inuence on nematode diversity in deep-sea environments (Gallucci et al., 2008b), but the more severe disturbance associated with mining may have pronounced and lasting impacts (Miljutin et al., 2011). Supplementary data to this article can be found online at http:// dx.doi.org/10.1016/j.jembe.2012.11.015. Acknowledgments Funding was provided by FRST through a postdoctoral fellowship to D. Leduc (UOOX0909), the programme Coasts & Oceans OBI (C01X0501), and the Department of Marine Science, University of Otago. We are thankful to Scott Nodder (NIWA) for facilitating sample collection, Keith Probert (University of Otago) for his support and guidance, and Anna Lawless (University of Waikato) for her help with processing of multicorer samples onboard RV Tangaroa. We also acknowledge the other participants of voyages TAN1103, the ofcers and crew of RV Tangaroa, and staff at NIWA's Mahanga Bay facilities for their technical support. We would like to thank J. Ingels for constructive criticisms on an early draft of the manuscript. [SS] References
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