Environ Monit Assess (2012) 184:1773–1779DOI 10.1007/s10661-011-2077-8

Bioaccumulation of heavy metals in the wolf spider,Pardosa astrigera L. Koch (Araneae: Lycosidae)

Myung-Pyo Jung · Joon-Ho Lee

Received: 9 September 2010 / Accepted: 11 April 2011 / Published online: 5 May 2011© Springer Science+Business Media B.V. 2011

Abstract Previous studies have proposed thatPardosa astrigera L. Koch (Lycosidae) can be usedas a biological indicator of heavy metal conta-mination in soil. In this study, we estimated thebioaccumulation levels and the bioconcentrationfactors (BCF) of four heavy metals (Cd, Cu, Pb,and Zn) in adult female P. astrigera collectedfrom various field sites according to heavy metalcontent gradient and broods. The relationship be-tween heavy metal content in the soil and thatin spiders was different depending on the heavymetals and the broods. However, heavy metalcontent in P. astrigera increased with increasingheavy metal content in the soil. While the heavymetal content in the soil was in the order ofZn > Pb > Cu > Cd, its content in P. astrigera was

M.-P. Jung · J.-H. Lee (B)Entomology Program, Department of AgriculturalBiotechnology, Seoul National University,Kwanak-ro 599, Shillim 9-dong, Kwanak-gu,Seoul, 151-921, South Koreae-mail: jh7lee@snu.ac.kr

J.-H. LeeResearch Institute for Agriculture and Life Sciences,Seoul National University, Kwansk-ro 599,Kwanak-gu, Seoul, 151-921, South Korea

Present Address:M.-P. JungNational Academic of Agricultural Science, RDA,Suwon, 441-707, South Korea

in the order Zn > Cu > Cd > Pb. The BCF for Cdin both of the broods was distinctly higher thanthose of the other heavy metals evaluated. Theseresults indicate that P. astrigera may be useful as abiological indicator of Cd soil contamination.

Keywords Pardosa astrigera · Heavy metals ·Bioaccumulation · Bioconcentration factor

Introduction

Ground-dwelling spiders are strong metal accu-mulators (Maelfait 1996; Marc et al. 1999; Junget al. 2007), and metal concentrations in theirbodies are considered an accurate reflection ofthe amount of metal in soil (Larsen et al. 1994;Maelfait 1996). Therefore, the analysis of heavymetal concentrations in ground-dwelling spiderscan allow for field evaluation of the heavy metalcontamination levels in ecosystems.

Wolf spiders accumulate more heavy metals intheir systems than do other ground-dwelling spiderspecies because of their frequent hunting activities(Marczyk et al. 1993; Rabitsch 1995; Wilczek andMigula 1996; Wilczek and Babczynska 2000). Pre-vious laboratory (Jung et al. 2005) and field (Junget al. 2007, 2008) experiments have suggested thatthe wolf spider, Pardosa astrigera L. Koch, is a po-tential bioaccumulator of heavy metals. P. astrig-era is the most abundant ground-dwelling spider

1774 Environ Monit Assess (2012) 184:1773–1779

in Korea and is distributed throughout Korea,Japan, China, Taiwan, and Russia. This spiderhas two broods, with first and second genera-tion adults appearing mostly in April–May andAugust–September, respectively.

In the present study, we measured the contentsof Cd (cadmium), Cu (copper), Pb (lead), and Zn(zinc), which are of great environmental concern,in the soil around and the bodies of adult femaleP. astrigera collected from field populations. Weevaluated the accumulation levels of these heavymetals in wolf spider females and compared the

bioconcentration factors of the four heavy metalsin the two levels, which are contamination level insoil and two broods.

Materials and methods

Study sites

Study sites were selected according to expecteddifferences in heavy metal contamination levels;their locations are shown in Fig. 1. Sampling was

IJ

PC

JC

GS

USJY

GYHS

JE

JH

SA

DH

BW

UW

GA

AS

GP1

GP2

Fig. 1 Locations and sampling date of study sites. ASAnsan (E 126◦49′, N 37◦18′; May and August 2006), BWBanweol (E 126◦53′, N 37◦19′; May and August 2006), DHDonghwa (E 126◦55′, N 37◦14′; May and August 2006,May and August 2007; May 2008), GP1 Gapyeong 1 (E127◦29′, N 37◦56′; August 2006, May and August 2007),GP2 Gapyeong 2 (E 127◦33′, N 37◦52′; August 2006, May2007, May and August 2008), GS Gasan (E 128◦33′, N36◦03′; May and August 2008), GY Gwangyang (E 127◦43′,N 34◦55′; May and August 2008), GA Gwanak (E 126◦57′,N 37◦27′; May and August 2008), HS Hwasun (E 127◦10′,

N 35◦04′; May and August 2008), IJ Inje (E 128◦14′, N37◦52′; May and August 2008), JC Jecheon (E 128◦10′, N37◦07′; May and August 2008), JE Jeongeup (E 126◦48′, N35◦36′; May and August 2007, May and August 2008), JHJanghang (E 126◦40′, N 36◦00′; May and August 2007, Mayand August 2008), JY Jinyeong (E 128◦43′, N 35◦17′; Mayand August 2008), PC Pyeongchang (E 128◦26′, N 37◦30′;May and August 2008), SA Songak (E 126◦46′, N 36◦56′;May and August 2008), UW Uiwang (E 126◦59′, N 37◦20′;May and August 2008), US Ulsan (E 129◦19′, N 35◦25′;May and August 2008)

Environ Monit Assess (2012) 184:1773–1779 1775

conducted in eight, nine, and 29 sites in 2006, 2007,and 2008, respectively. Sampling was performedtwice (May and August) a year at the same site.

Spider and soil sampling

Spiders and soil samples were collected on thesame date. For spider sampling, only female adultspiders with an egg sac were collected by handto avoid differences in metal content due toage and/or sex (Williamson 1980; Rabitsch 1995).More than 15 spiders were collected at each sam-pling site. For soil sampling, top soil samples (ca.500 g each) were collected from three differentlocations (ca. 10 m apart between samples) in eachsite from which spiders were collected.

Chemical analysis

Spiders collected in each sampling site were di-vided into three groups with five to ten individ-uals. These groups were used as a replicate. Spi-ders were oven-dried for 2 h at 135–150◦C andweighed. They were then digested with 2 ml of1 N HNO3 solution until no solid material wasobserved in the solution. These solutions were di-luted 100-fold with 10 ml of 1 N HCl and distilledwater after being cooled to room temperature.

Soil samples were dried for 7 days at roomtemperature (Nadal et al. 2004). The dried soilsamples (10 g) were mixed with 50 ml of 0.1 N HCland HNO3 solution, shaken for 1 h at 30◦C, andthen filtered. The filtrate was diluted tenfold with0.1 N HCl and distilled water.

The heavy metal (Cd, Cu, Pb, and Zn) concen-trations in all prepared soil and spider samples

were analyzed using inductively coupled plasmamass spectrometry.

Data analysis

Data from sites where heavy metals were notdetected were excluded in the analysis. Pearson’scorrelation analysis was conducted to determinerelationships between soil and body heavy metalcontent in each sampling period using the Rpackage (R Development Core Team 2009). Fur-thermore, to evaluate metal bioaccumulation insoil, bioconcentration factors (BCF) were calcu-lated (Hussein et al. 2006). In this study, BCFwas defined as the ratio of the metal concen-tration in spiders to that in the soil. The heavymetal contents and the BCFs for the four heavymetals were compared between two broods us-ing t test and the BCFs among the four heavymetals were compared using Tukey’s test imple-mented in the R package (R Development CoreTeam 2009).

Results

The heavy metal contents in the soil samples andspiders are shown in Table 1. Overall, the meanheavy metal content in the soil was highest inthe order of Zn, Pb, Cu, and Cd in both of thetwo broods. In P. astrigera, Zn was also highest,followed by Cu, Cd, and Pb. Heavy metal contenttended to be high in the autumn brood than in thespring brood. Especially, Cd content in soil (t =2.47, p = 0.0166) and body (t = 2.79, p = 0.0072)and Pb content in body (t = 3.01, p = 0.0040) in

Table 1 Heavy metal content in soil (mg/kg dw; mean ± SE) and body (μg/g dw; mean ± SE) of P. astrigera

Type Broods Cd Cu Pb Zn

Soil Spring 0.22 ± 0.058 25.03 ± 4.183 28.53 ± 3.212 83.44 ± 10.090*Autumn 0.40 ± 0.038* 25.79 ± 1.992 34.02 ± 3.919 50.98 ± 6.099Total 0.31 ± 0.037 25.65 ± 2.403 31.13 ± 2.514 68.46 ± 6.350

Body Spring 6.77 ± 0.984 154.45 ± 8.026 1.52 ± 0.209 208.40 ± 23.818Autumn 10.94 ± 1.128** 188.07 ± 17.483 7.83 ± 2.206** 166.04 ± 48.468Total 8.72 ± 0.778 169.37 ± 9.380 4.50 ± 1.123 185.53 ± 25.408

Asterisks marks mean the significant difference of heavy metal content in soil or body between spring brood and autumnbrood*0.01 < p < 0.05; **0.001 < p < 0.01

1776 Environ Monit Assess (2012) 184:1773–1779

the autumn brood were significantly higher thanthose in the spring brood.

The relationship between heavy metal contentin the soil and that in spiders was different de-pending on the heavy metals and the broods(Fig. 2). However, the overall heavy metal con-centration in spiders increased with increasingsoil content of heavy metals; for example, thehighest correlation was seen between the levelsof Cd in the soil and those in spiders in thespring brood (r = 0.76, p < 0.001). The relation-ship between Cd and Cu content in soil and inspiders was stronger in the spring brood than in

the autumn brood while it of Pb and Zn was thereverse.

The bioconcentration factors (mean ± SE) ofheavy metals, from highest to lowest, were Cd >

Cu > Zn > Pb. The BCF of Cd was significantlyhigher than those of the other heavy metals inboth of the two broods (spring brood: df = 3,F = 68.34, p < 0.0001; autumn brood: df = 3, F =6.07, p = 0.0008; Fig. 3). The BCF value was notsignificantly different between the broods for Cd,Cu, and Zn. Although the BCF value of Pb wasvery low, it was higher of the autumn brood thanit was of the spring brood (t = 3.11, p = 0.0030).

Cd content in soil (mg/kg)

Cd

co

nte

nt

in s

pid

ers

(µg

/g)

0

5

10

15

20

25

30

r=0.02

r=0.51***

r=0.76***

Cu content in soil (mg/kg)

Cu

co

nte

nt

in s

pid

ers

(µg

/g)

0

100

200

300

400

500

r=0.17

r=0.32*

r=0.61***

Pb content in soil (mg/kg)

Pb

co

nte

nt

in s

pid

ers

(µg

/g)

0

10

20

30

40

50

60

r=0.36

r=0.31*

r=0.20

Zn content in soil (mg/kg)

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 0 20 40 60 80 100 120

0 20 40 60 80 100 120 0 50 100 150 200 250 300 350

Zn

co

nte

nt

in s

pid

ers

(µg

/g)

0

200

400

600

800

1000

r=0.32

r=0.41**

r=0.64***

A) B)

C) D)

Fig. 2 a–d Relationships between heavy metal content(milligrams per kilogram dry weight) in soil samples andspiders. Long dash line (closed circles), dash line (open cir-cles), and solid line mean data from spring brood, autumnbrood, and all brood, respectively. The stars indicate that

the heavy metal content in the spiders was significantlycorrelated with the heavy metal content in the soil atsignificance levels of ***p < 0.001; **0.001 < p < 0.01;*0.01 < p < 0.05

Environ Monit Assess (2012) 184:1773–1779 1777

S

BC

F v

alu

e o

f C

d

0

20

40

60

80

BC

F v

alu

e o

f C

u, P

b, Z

n

0

2

4

6

8

10

12

**A S S SA A A

CdCu

Pb

Zn

Fig. 3 The BCF values for each heavy metal. S springbrood, A autumn brood, **0.001 < p < 0.01

Discussion

It is well known that biotic factors such as food,sex, and age structure as well as abiotic factorssuch as soil properties can affect the metal contentin organisms. In our study, the high metal contentin the autumn brood in P. astrigera may relate tothe high metal content in soil rather than otherfactors (Table 1).

In terrestrial invertebrates, the relationship be-tween total heavy metal soil content and the in-ternal metal content in invertebrates tends to bestrong in the order of Pb > Cd > Cu, Zn (Heikenset al. 2001). It has been speculated that in ter-restrial invertebrates Cu and Zn can be regulatedto a certain degree, resulting in a constant bodyconcentration over a range of soil contents (Beyeret al. 1985; Morgan and Morgan 1988; Van Gestelet al. 1993; Heikens et al. 2001; Hussein et al.2006; Del Toro et al. 2010), while Cd and Pb,non-essential metals, are not regulated (Van Gestelet al. 1993; Heikens et al. 2001; Hussein et al. 2006).

Heavy metal concentrations in P. astrigera in-creased significantly with increasing soil concen-trations, but the pattern of the relation differedfrom those seen in other terrestrial invertebrates.The pattern for the correlation in P. astrigera was:Cd > Zn, Cu > Pb. The BCF values followed thissame trend, but the BCF for Cd was significantlyhigher than those of the other heavy metals, indi-cating that P. astrigera is an effective accumulatorof Cd in both of the two broods.

Many ground-dwelling spiders either do notaccumulate Pb or accumulate Pb at very low levels(Hopkin and Martin 1985; Clausen 1989; Larsenet al. 1994; Wilczek et al. 2004; Jung et al. 2005,2007), consistent with our results. P. astrigeraaccumulated Cd more selectively than they didHg and Pb in laboratory experiments (Jung et al.2005) and accumulated high levels of Cd in fieldstudies (Jung et al. 2007).

In the present study, although lower BCF val-ues were observed compared to those of a previ-ous study (BCFCd, 149.7; BCFPb, 9.3) (Jung et al.2007), the BCF for Cd was still much higherthan that for Pb, consistent with the previousstudy.

The high BCF of Cd in spiders could be re-sponsible for the high toxicity of Cd (Devkota andSchmidt 2000). Storage of Cd in spiders is one ofthe mechanisms for detoxification of heavy metals(Maelfait 1996). Spiders have two types of metal-containing granules in the digestive cells of thehepatopancreas of spiders that are renewed within24 h and eliminated via the feces (Hopkin 1989).Lead and Zn are stored in type A granules, whichare amorphous deposits of calcium phosphates,whereas Cd and Cu are stored in type B granules,which are originated from the lysosomal systemand contain mainly acid phosphatase. This mayexplain the relatively higher BCFs of Cd and Cucompared to those of Pb and Zn.

The long biological half-life and high solubilityof Cd might be also responsible for the higherbioaccumulation of this heavy metal compared tothose of Pb (Van Hook and Yates 1975; Roth-Holzapfer 1990). Cd has higher solubility and isless stable in compound form than Pb (Weng et al.2002). Total soluble Cd is also correlated withtotal metal content of the soil and strongly relatedto soil pH. However, the solubility of Pb is lessdependent on the soil properties (McBride et al.1997).

Overall, the accumulation patterns and BCFvalues that we measured for P. astrigera indicatethat this spider can accumulate Cd to a greaterdegree than it can other heavy metals. Thus, P.astrigera is a useful accumulation indicator of en-vironmental Cd contamination.

1778 Environ Monit Assess (2012) 184:1773–1779

Acknowledgements This work was supported by a grantfrom the Ministry of Environment (091-061-043), and fund-ing from the Brain Korea 21 program.

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