Ecotoxicology and Environmental Safety 156 (2018) 452–462

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Ecotoxicology and Environmental Safety

journal homepage: www.elsevier.com/locate/ecoenv

Review

Bioaccumulation of cadmium in soil organisms – With focus on wood ash T

application
⁎
Louise Hindborg Mortensena, Regin Rønna,b,c, Mette Vestergårda,d,
a
Terrestrial Ecology Section, Department of Biology, University of Copenhagen, Universitetsparken 15, DK-2100 Copenhagen Ø, Denmark
b
Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China
c
Arctic Station, University of Copenhagen, Qeqertarsuaq, Greenland
d
Department of Agroecology, Aarhus University, Forsøgsvej 1, DK-4200 Slagelse, Denmark

A R T I C LE I N FO A B S T R A C T

Keywords: Harvesting whole-tree biomass for biofuel combustion intensifies removal of nutrients from the ecosystem. This
Heavy metal accumulation can be partly amended by applying ash from the combustion back to the system and thus recycle the nutrients.
Cadmium biomagnification However, besides being rich in inorganic nutrients, ash also contains trace amounts of heavy metals. Due to the
Soil fauna risk of toxic effects and trophic transfer of heavy metals, especially cadmium, legislation usually restricts the use
Cadmium availability
of ash as a soil amendment.
Biofuel
In order to provide researchers and governmental agencies with a tool to assess the risk of cadmium bioac-
cumulation in specific soil systems after ash application, we review: 1) the properties of ash; 2) the chemical and
toxic properties of cadmium; 3) the key factors affecting cadmium bioavailability, cadmium uptake-, storage-
and elimination-abilities in soil organisms and the risk of cadmium accumulation and biomagnification in the
soil food web; 4) how ash impact on soil can change the risk of cadmium bioaccumulation.
We conclude that for assessing the risk of cadmium bioaccumulation for specific sites, it is necessary to
consider both the type and composition of ash, the soil conditions and organism composition on the site. On a
general basis, we conclude that granulated ashes low in cadmium content, applied to low pH soils with high
organic matter content, in systems with low abundances of earthworms, isopods and gastropods, will have a low
risk of cadmium accumulation.

1. Introduction maximum threshold for cadmium (Cd) in ash application to 60 g Cd

Burning of fossil fuels and greenhouse gas emissions are the major
causes for current global climate change. Therefore, the European
Council in 2006 established a mandatory target of 20% renewable en-
ergy of the total energy consumption by 2020. Part of the use of fossil
fuels is replaced by burning of biomass, e.g. wood chips, for heating and
electricity (COM, 2006). Harvesting whole-tree biomass from the forest
intensifies nutrient removal from the ecosystem, compared to tradi-
tional harvest of timber (Raulund-Rasmussen et al., 2008). Burning of
wood generates up to 1% ash by weight and ash is thus a significant by-
product from biofuel power plants (Pitman, 2006). Disposal of ash at
waste disposal sites is costly (Ingerslev et al., 2011), but as forest fer-
tilizer, ash can help compensate for the removal of biomass for biofuel
production (Swedish National Board of Forestry, 2002). Wood ash
contains high amounts of plant nutrients, but also trace amounts of
heavy metals (Pitman, 2006), which generally sets the limits for the use
of ash as a fertilizer. The legislative restriction in Denmark sets the
ha−1 75 year−1 for forest systems and 0.8 g Cd ha−1 year−1 for farm-
land (Danish Ministry of the Environment, 2008). Several reviews have
covered research of wood ash effects on forest soil ecosystems (Augusto
et al., 2008; Huotari et al., 2015; Pitman, 2006; Vance, 1996) and
toxicity and accumulation of heavy metals in organisms (Anderson
et al., 2013; Bengtsson and Tranvik, 1989; Heikens et al., 2001), but
very little has been published on the combination of wood ash appli-
cation and Cd bioaccumulation (Perkiömäki and Fritze, 2005). Because
many of the legislative restrictions for wood ash application are
grounded in the accumulation risk of heavy metals (Danish Ministry of
the Environment, 2008), and because biomass burning and hence ash
production is increasing, it is highly relevant to elucidate the risk of
terrestrial Cd bioaccumulation after wood ash application.
The aim of this review is to make a detailed analysis of the risk of Cd
accumulation in soil organisms after ash application to soils. To achieve
this, we review: 1) the properties of ash; 2) the chemical and toxic
properties of Cd; 3) the physical and biological factors affecting the risk

⁎
Corresponding author at: Terrestrial Ecology Section, Department of Biology, University of Copenhagen, Universitetsparken 15, DK-2100 Copenhagen Ø, Denmark.
E-mail addresses: louisem@bio.ku.dk (L.H. Mortensen), rmr@science.ku.dk (R. Rønn), mevmadsen@bio.ku.dk (M. Vestergård).

https://doi.org/10.1016/j.ecoenv.2018.03.018
Received 29 August 2017; Received in revised form 23 February 2018; Accepted 6 March 2018
0147-6513/ © 2018 Elsevier Inc. All rights reserved.
L.H. Mortensen et al.
of Cd accumulation in soil organisms; 4) how the ash impact on soil can
change the risk of cadmium bioaccumulation. Lastly, we will combine
this to elucidate which specific factors should be assessed before
amending a soil system with ash, in order to minimize the ecological
risk of Cd in the system. Our hope is that the conclusions and synthesis
will provide researchers, governmental agencies and forest managers a
tool to assess Cd bioaccumulation risk in their specific targeted soil
system.
2. Application of wood ash
Acidification is a common problem in production forestry. Wood
ash is generally highly alkaline and the neutralizing effect is probably
the most important characteristic of ash amendment to soil, but the
effect depends on ash quantity and quality, as well as the site. Soils with
intrinsically low pH and low organic matter content have shown the
highest pH increases (Demeyer et al., 2001). For humus and peat sur-
faces pH increases between 0.5 and 3 pH units have been reported
depending on the ash dose (Huotari et al., 2015).
Because ash contains valuable plant nutrients, it is also an efficient
fertilizer (Demeyer et al., 2001). The major components of wood ash
are calcium (Ca), potassium (K), magnesium (Mg) and phosphorus (P).
Other macro-elements are aluminium (Al), iron (Fe), sodium (Na),
manganese (Mn) and sulphur (S). Due to evaporation during combus-
tion, all ashes are very low in nitrogen (N) (Demeyer et al., 2001;
Etiégni and Campbell, 1991; Ingerslev et al., 2011; Karltun et al., 2008).
Thus, the efficiency of ash as a fertilizer depends on the existing N in the
system and/or the rate of N deposition. Fertilization with wood ash has
shown positive effects on tree production at sites, which receive N
deposition or are rich in organic matter, like peat soils. However, no
positive effects are seen on e.g. mineral soils where N is the limiting
nutrient (Huotari et al., 2015; Karltun et al., 2008). In spite of the
negligible N content in wood ash, the increased pH can indirectly in-
crease microbial activity and thus N mineralization and availability
(Genenger et al., 2003; Jäggi et al., 2004; Martikainen, 1985; Weber
et al., 1985; Vestergård et al., 2017).
Wood ash also consists of a range of trace elements that originate
from the incinerated wood. These include arsenic (As), silver (Ag),
molybdenum (Mo), mercury (Hg), nickel (Ni), vanadium (V), zinc (Zn)
and cadmium (Cd) (Demeyer et al., 2001; Etiégni and Campbell, 1991;
Karltun et al., 2008). Some of these are micro-nutrients; some are non-
essential heavy metals (Huotari et al., 2015). Because ash usually ori-
ginates from trees grown on a larger area than the area where the ash is
used as a fertilizer, the heavy metals from the wood ash will con-
centrate in the amended area. This can increase the total heavy metal
content in the soil (Huotari et al., 2015). The biggest concern regarding
ash amendment is the heavy metal content, particularly Cd (Beyer,
2000; Saarsalmi et al., 2001).
3. Toxicity of Cd
Cd taken up by plants or other organisms is incorporated into bio-
logical systems, and can be transferred to higher trophic levels via
consumption. Due to loss of biomass during respiration and in excretion
products, there is a risk of bioaccumulation at higher trophic levels
(Janssen et al., 1993). Increasing Cd concentrations in consumers can
have detrimental effects, as seen during the first half of the 20th century
in Japan, where Cd poisoning caused osteomalacia (severe pain and
fracture of bones and joints) in humans after consumption of Cd con-
taminated rice (Nordberg, 2009).
Because Cd mobility is high compared to other heavy metals
(Cd > Ni > Zn > Mn > Cu > Pb=Cr > Hg (Brümmer, 1986; Kim
et al., 2015)) it has a high potential for bioaccumulation in food webs.
Cd resembles the essential metals Zn and Cu and may be taken up along
the same pathways as Zn and Cu (Tyler et al., 1989). As a non-essential
metal, regulation is probably either less pronounced or absent for Cd
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Ecotoxicology and Environmental Safety 156 (2018) 452–462
(Janssen and Hogervorst, 1993; Van Gestel et al., 1993). With no or
little regulation, Cd can ‘hitchhike’ into the cells and accumulate in the
organism. Cd can also replace Zn in metabolic processes, which is one
of its toxic actions in the cell (Qiu et al., 2011; Rani et al., 2014). Ad-
ditionally, Cd causes indirect oxidative stress, which can lead to DNA
damage, inhibit DNA repair mechanisms and inhibit apoptosis (Rani
et al., 2014).
In soil organisms this has been expressed as decreased growth
(Bengtsson and Tranvik, 1989; Posthuma et al., 1992; Russell et al.,
1981; Swaileh and Ezzughayyar, 2000; Van Straalen et al., 2005),
prolongation of the juvenile period (Russell et al., 1981), decreased
reproduction (Anderson et al., 2013; Bengtsson and Tranvik, 1989;
Russell et al., 1981; Scheirs et al., 2006) and increased mortality (Qiu
et al., 2011). On an ecosystem scale, Cd toxicity has disrupted ecolo-
gical processes by affecting decomposition (Vig et al., 2003), changing
community structure (Khalil et al., 2009) and reducing biomass (Tyler
et al., 1989), diversity (Lodenius et al., 2009; Tyler et al., 1989) and
relative abundance of species (Lodenius et al., 2009). The concentration
of Cd can also increase in higher trophic levels, e.g. in birds
(Roodbergen et al., 2008) or mice (Hunter et al., 1987b), which can
export Cd from the contaminated system and disrupt the ecological
functioning of other ecosystems. Furthermore, it is at risk of entering
the human food chain.
4. Bioaccumulation of Cd in soil organisms
Janssen et al. (1993) defines bioaccumulation as the concentration
increase of a substance in an organism compared to its environmental
concentration through any kind of uptake process. Predicting how ash
will influence Cd bioaccumulation in a soil system is difficult, but
bioaccumulation is often used as a parameter when assessing the risk of
toxicants in the environment (McGeer et al., 2003). The Cd con-
centration in soil/litter/ash cannot accurately predict the concentration
in soil organisms (Bengtsson and Tranvik, 1989). However, by con-
sidering relevant factors, an estimate of potential bioaccumulation after
ash application can be made. Firstly, we need to identify the most im-
portant physicochemical factors in the soil that influence Cd binding
properties and bioavailability. Secondly, we need to address the factors
that determine Cd accumulation in the soil organisms. The following
will summarize and pinpoint the key factors that generally influence
bioaccumulation of Cd in soil organisms – irrespective of ash addition.
4.1. Physical factors affecting Cd bioavailability and Cd uptake in
organisms
Although often used, total content of heavy metal in a soil is not the
best measure for contaminant exposure to soil dwelling organisms
(Lanno et al., 2004; Plette et al., 1999; Sauvé et al., 2000b). Only part of
the heavy metal present in soil is available for uptake and incorporation
into the organisms. The metal must be bioavailable and not irreversibly
bound to the soil matrix (Blume and Brümmer, 1991; Brümmer, 1986;
Kim et al., 2007; Lanno et al., 2004). We define the bioavailable frac-
tion as that part of the total amount of Cd that can be sorbed by an
organism in a terrestrial system.
Many studies investigating bioavailability in soil focus on plant
availability (Blume and Brümmer, 1991; Brown et al., 2004; Gall et al.,
2015; Kirkham, 2006; McLaughlin et al., 1998; Scheifler et al., 2003;
Wang et al., 2010). Cd is most readily available to plants as Cd++,
whereas plants take up CdCl+ more slowly and do not take up Cd-
humate (Crea et al., 2013).
Several different methods have been used to evaluate bioavailability
(Brümmer, 1986; Lanno et al., 2004; Yin et al., 2014), for example soil
concentration of dissolved metals (Blume and Brümmer, 1991; Brown
et al., 2004; Christensen, 1989; Kim et al., 2015; Paradelo et al., 2011),
metal partitioning coefficient – the ratio between soluble and total
metal concentration in the soil (Anderson and Christensen, 1988;
L.H. Mortensen et al. Ecotoxicology and Environmental Safety 156 (2018) 452–462

Table 1
Summary of soil factors influencing bioaccumulation of cadmium. Bold signifies factors to test before (and after) ash amendment.
Physicochemical soil factor Effect Causes

pH
Ca concentration
Cl concentration
Organic matter content
↓ pH = ↑ Cd bioavailability
↑ pH = ↓ Cd bioavailability
↑ Ca = ↑ Cd bioavailability
↑ Cl = ↑ Cd bioavailability
↑ Organic matter = ↓ Cd
bioavailability
↓ pH = ↑ [H+] = ↓ net negative surface charge of soil complexes → ↑ cation exchange with Cd++ → ↑ Cd++
availability in the soil matrix increases
↑ pH = ↓ [H+] = ↑ net negative surface charge of soil complexes → Cd++ is absorbed onto less accessible
sites of humus particles → Cd++ availability in the soil matrix decreases
Ca++ competes with Cd++ for the same binding sites on soil particles
↑ Cl- = ↑ CdCl ion pairs → small, inorganic available compound
↑ Organic matter = ↑ negatively charged complexes → Cd++ is absorbed to less accessible sites of humus
particles → Cd++ is less available in the soil matrix

Janssen et al., 1997; Lee et al., 1996; Qiu et al., 2011; Sauvé et al.,
2000a), metal speciation – different forms of a metal have different
solubility (Kim et al., 2015; Sauvé et al., 2000b; Wang et al., 2010),
model calculations (Plette et al., 1999) or internal metal concentration
of the studied organism (Crommentuijn et al., 1994; Lock and Janssen,
2001; Peijnenburg et al., 1999; Van Straalen et al., 2005; Veltman et al.,
2008).
In spite of these different methods, studies agree that besides total
Cd content in the soil, pH is the most important soil physicochemical
parameter for Cd bioavailability (Anderson and Christensen, 1988;
Anderson et al., 2013; Christensen, 1989; Janssen et al., 1997; Lee
et al., 1996; Ma et al., 1983; Pauget et al., 2011; Peijnenburg et al.,
1999; Plette et al., 1999; Sauvé et al., 2000b).
4.1.1. pH
At low pH, Cd ions bind non-specifically to soil particles and are
relatively easily replaced by H+ via cation exchange. Hence, Cd is so-
luble in the soil matrix at low pH. As pH increases, net negative surface
charge of soil complexes increases. Thus, Cd++ can be adsorbed to less
accessible sites of clay and humus particles (Crea et al., 2013; Kim et al.,
2015). Sauvé et al. (2000b) found that approximately 75% of the total
Cd pool was free Cd++ and inorganic Cd (e.g. CdCl) at pH 3. That
fraction had dropped to 35% at pH 6. At pH 8, the free Cd++ and
inorganic Cd had increased to approximately 55% due to formation of
hydrolysis and carbonate ion-pairs. It should be noted that pH 8 is
unusually high for soils. Even after treatment with 90 t ash ha−1, we
found a mean pH of 7.7 ± 0.12 in a Norway spruce plantation with a
pH of 3.83 ± 0.08 in unamended control plots (Vestergård et al.,
2017).
4.1.2. Cations
Other cations also compete with Cd++ for binding sites on soil
particles. Especially soils with high Ca++ concentrations can increase
the availability of Cd (Christensen, 1984; Escrig and Morell, 1998;
Plette et al., 1999). The presence of Cl- increases Cd concentration in
plants (Brümmer, 1986; Escrig and Morell, 1998; McLaughlin et al.,
1998), because the formation of CdCl ion pairs decreases the adsorption
of Cd to soil particles (Sauvé et al., 2000b). Although Ca++ and Cl-
influence Cd availability, they hold no significance compared to pH and
organic matter in the system (Christensen, 1989).
4.1.3. Organic matter
Soil organic matter content is important for Cd bioavailability
(Christensen, 1989; Lee et al., 1996; Pauget et al., 2011; Sauvé et al.,
2000b). Organic matter holds a range of negatively charged functional
groups (Kim et al., 2007) and again due to cation exchange, Cd++ can
bind very efficiently to organic matter, even at very low pH (pH 3–4)
(Brümmer, 1986).
Soils with high pH and high organic matter content thus bind Cd
strongly. Cd bound in large complexes is believed to be non-available to
plants because they are unable to cross the cell membrane of roots
(Wang et al., 2010). Smaller, inorganic complexes, such as CdCl, CdOH
and CdHCO3, are potentially bioavailable for plant uptake (Kim et al.,
2015).
However, some soil feeding organisms, notably earthworms and
gastropods consume soil and organic matter (Chevalier et al., 2001;
Elmslie, 1998), where metals could be bound (Scheifler et al., 2003).
Consequently, it can be important to distinguish between bioavail-
ability for plants and soil organisms. Scheifler et al. (2003) showed that
a significant 16% of the Cd content in the snail Helix aspersa originated
from the “non-available pool” of soil Cd. Cd strongly bound to the soil
particles can thus be taken up by soil organisms and potentially be
transferred to and concentrated in higher trophic levels. For earth-
worms that ingest organic matter, the percentage of “non-available”-
available Cd may be much more significant. It is thus important to
consider the technique for measuring metal contamination for mon-
itoring Cd bioaccumulation after ash application.
Table 1 summarizes the effects and causes of soil biochemical fac-
tors on Cd bioaccumulation, as well as recommendations for soil factors
to test before (and after) ash amendment.
4.2. Organism characteristics of importance for bioaccumulation
Bioaccumulation in invertebrates does not correlate with body size
or biomass (Morgan and Morgan, 1988; Van Straalen and Van Wensem,
1986), trophic level (Roberts and Johnson, 1978; Van Straalen and Van
Wensem, 1986) or taxonomic group (Andersen, 1979; Ireland, 1979).
Thus, the potential for Cd accumulation does not only differ between
taxonomic groups and species (Bengtsson and Tranvik, 1989), but is
also unpredictable judged on the basis of common physiological mea-
sures (Janssen et al., 1993). Instead, it is the species-specific factors
(Rainbow, 2002; Tyler et al., 1989), such as history, morphology and
physiology that influence the uptake, storage and elimination of a
toxicant, and hence bioaccumulation (Bengtsson and Tranvik, 1989;
Newman, 2003). In the following we will review these factors for a
range of soil organisms to pinpoint organisms or organism groups that
are most prone to accumulate Cd and thus, most important to monitor
after ash application.
4.2.1. Uptake of Cd
Uptake of Cd depends on exposure intensities and uptake routes.
The exposure varies with life history characteristics, which to some
degree also determines uptake routes. Uptake of metals include in-
halation and transport over the outer cell wall, either epidermal or after
ingestion. However, inhalation is probably of minor importance for soil
organisms (Janssen et al., 1993). Uptake through the cell wall/epithe-
lium depends on morphology, diet and physiology (Anderson et al.,
2013; Heikens et al., 2001; Korthals et al., 1996; Nannoni et al., 2011;
Son et al., 2011; Suthar et al., 2008).
4.2.1.1. Exposure and Life History. Organisms with long generation
times have long exposure time (Bengtsson and Tranvik, 1989;
Bongers and Ferris, 1999) and are thus presumably more susceptible
to contaminants. Cd elimination often occurs slowly, as seen in snails
(Williamson, 1980), earthworms (Honda et al., 1984; Veltman et al.,
2007), isopods, millipedes (Crommentuijn et al., 1994) and mammals

454
L.H. Mortensen et al.
(Neathery and Miller, 1975). Prolonged exposure consequently
increases Cd concentration in the individual organism. For instance,
Cd concentration increases with age for the terrestrial snail Cepaea
hortensis (Williamson, 1979). Development stage can also influence Cd
exposure, because habitat and food preferences can differ considerably
between development stages, e.g. insect larvae and imago stages.
Hunter et al. (1987a) showed that Coleoptera larvae that are in direct
contact with the contaminated litter had a higher Cd concentration than
adult individuals, which are not directly exposed to the litter.
4.2.1.2. Surface uptake. Direct contact to the contaminated
environment intensifies the exposure and the relative contact to the
environment is greater for organisms with a large surface-to-volume
ratio (Ray et al., 1980; Smock, 1983; Tyler et al., 1989). Although
rarely considered, microorganisms are immersed in the substrate and
have a high surface-to-volume ratio. Furthermore, they often have a
negatively charged cell wall, which makes them prone to cation, e.g.
Cd, uptake. Microorganisms are therefore very prone to Cd
accumulation (Gall et al., 2015). Because microorganisms are a
substantial part of the diet for many soil organisms, this should not
be neglected in Cd bioaccumulation risk analyses of soil organisms.
For invertebrates, especially soft-bodied organisms such as en-
chytraeids (Didden and Römbke, 2001), nematodes (Kammenga et al.,
1994; Korthals et al., 1996) and earthworms (Oste et al., 2001; Saxe
et al., 2001; Van Gestel et al., 1993; Veltman et al., 2007; Vijver et al.,
2003), but also collembolans (Vijver et al., 2006b), epidermal uptake
plays a major role in Cd uptake. For earthworms, uptake has been
shown to correlate to the soluble metal concentration in the medium
(Oste et al., 2001). However, for nematodes the differences in thickness
and collagen density of the ectodermal tissue influence permeability
and hence uptake of substances (Kammenga et al., 1994).
There is some debate, whether the main uptake route of heavy
metals in earthworms is dietary (Hobbelen et al., 2006; Morgan et al.,
2004; Nannoni et al., 2011; Oste et al., 2001; Stürzenbaum et al., 2004)
or epidermal (Hobbelen et al., 2006; Peijnenburg et al., 1999; Reinecke
et al., 1997; Saxe et al., 2001; Van Gestel et al., 1993; Vijver et al.,
2003). This debate is probably rooted in the fact that experiments have
been performed at highly variable soil conditions. Also, the relative
importance of physicochemical factors varies greatly between different
heavy metals and it is thus difficult to determine one main uptake route
for a range of metals. However, even when focusing on one heavy
metal, a small change in for example pH and organic matter content
will change the fractions of soluble and humic-bound Cd (see Table 1).
The soluble fraction is available for both dermal and dietary uptake,
whereas humic-bound Cd only is available for dietary intake. To our
knowledge, no studies have analysed if there is a correlation between
the physicochemical conditions and the uptake route of heavy metals in
earthworms. Such an investigation could potentially close the discus-
sion.
4.2.1.3. Dietary intake. The dietary intake of heavy metals is the most
important route for surface-living organisms, like insects and arachnids
(and vertebrates) (Janssen et al., 1993). Cd body concentrations of
detritivores and herbivores, feeding on directly contaminated organic
matter and/or plant material, have been shown to correlate with the
surrounding metal concentration (Nannoni et al., 2011; Rocco et al.,
2011; Suthar et al., 2008). Thus, total metal content and/or the
bioavailable fraction of metal in the soil is often used as a measure of
exposure, but for carnivorous or omnivorous organisms the dietary
intake of Cd may differ (Nannoni et al., 2011). Actual metal
composition and concentration in the diet of carnivores or omnivores
is often difficult to estimate (Janssen et al., 1993). For example, certain
spiders, like Centromerus sylvaticus, consume parts of their prey with the
highest metal concentrations (Hunter et al., 1987a; Van Straalen and
Van Wensem, 1986). Thus, feeding behaviour also influences dietary Cd
uptake. In fact, some organisms avoid contaminated food all together.
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Ecotoxicology and Environmental Safety 156 (2018) 452–462
In laboratory choice-experiments some soil and litter invertebrates (e.g.
earthworms, isopods and collembolans) discriminated between clean
and heavy metal contaminated food (Behmer et al., 2005; Jhee et al.,
1999; Pollard and Baker, 1997; Rathinasabapathi et al., 2007; Scheirs
et al., 2006; Tranvik and Eijsackers, 1989; Zidar et al., 2004) or soil
(Lukkari and Haimi, 2005; Sjögren et al., 1995). In the field however,
this is less relevant. Although the contamination/ash is probably
unevenly distributed, creating uncontaminated spots, the population
as a whole, would not be able to avoid contamination.
4.2.1.4. Digestive uptake. The physiology of the digestive system
determines metal uptake after ingestion. The uptake of nutrients and
other elements, like Cd, is influenced by the flowrate of food through
the digestive system (Pokarzhevskii and Van Straalen, 1996), the
absorption efficiency of the digestive system as well as properties of
the food (Tyler et al., 1989). The midgut is the main site of nutrient
absorption and hence heavy metal uptake of terrestrial invertebrates.
Centipedes and millipedes lack the midgut diverticulae or another large
organ for storage and absorption (Hopkin, 1989). Metal containing
granules have been observed in the faeces of the centipede Lithobius
variegatus (Hopkin et al., 1985), suggesting that the reduced digestive
system of centipedes and millipedes to some extent prevent uptake of
heavy metals (Bengtsson and Tranvik, 1989).
Also the chemical properties, especially pH, in the digestive system
can play a key role for heavy metal uptake. pH in the digestive tract can
differ from the environmental pH and heavy metals can thus become
more or less bound to the food after ingestion. In isopods, collembolans,
earthworms and most other detrivores the gut pH is within one unit of
neutrality. However, some parts of the digestive tract may be strongly
acidic or alkaline (Hopkin, 1989), which can facilitate the uptake of
heavy metals in the midgut (see Table 1).
4.2.1.5. Regulation of uptake. The requirements for essential elements
usually regulate the flow of food through the digestive system
(Pokarzhevskii and Van Straalen, 1996). Organisms with a high
demand for a specific essential metal have also been found to
accumulate the analogue non-essential metal to a higher degree than
other metals, which could pose a strong potential for Cd
bioaccumulation. Cd may be taken up along the same pathways as
the essential metals Zn and Cu (Tyler et al., 1989). Cd and Zn body
concentration correlated for nine species of arthropods, (Janssen and
Hogervorst, 1993), and earthworms (Nannoni et al., 2011) which
suggest that the need for Zn controls Cd uptake.
Zn uptake is often regulated in invertebrates (Ireland, 1979; Morgan
and Morgan, 1988; Nannoni et al., 2011; Peijnenburg et al., 1999;
Roberts and Johnson, 1978). Some studies suggest that Cd is, to some
degree, regulated in the same way as Zn (Hunter et al., 1987a; Van
Straalen and Van Wensem, 1986), whereas others found no evidence for
regulation (Andersen and Laursen, 1982a; Hunter et al., 1987a; Janssen
and Hogervorst, 1993; Van Gestel et al., 1993). However, this dis-
crepancy could be caused by ‘competition’ between the two metals.
This has been shown for lead (Pb), an analogue to calcium (Ca). At high
Ca concentrations in soil Ca outcompetes Pb for uptake into the or-
ganism and supresses Pb accumulation (Andersen, 1979; Andersen and
Laursen, 1982a; Ireland, 1979). However, in situations where Ca is in
demand, Pb accumulates to a higher degree (Bengtsson and Tranvik,
1989). Likewise, plant Cd uptake increases, when Zn availability is low
(Johansen et al., manuscript submittet), hence a similar mechanism
could occur in soil organisms. Organisms, like earthworms, which are
highly dependent on Zn for physiological processes, take up and store
Zn for future demand (Nannoni et al., 2011), which could pose a strong
potential for Cd bioaccumulation.
Table 2 summarizes the factors that principally determine Cd ex-
posure and uptake in individual organisms.
L.H. Mortensen et al. Ecotoxicology and Environmental Safety 156 (2018) 452–462

Table 2
Summary of organism uptake characteristics influencing bioaccumulation of heavy metals in general.
Factors Effect Cause

Life history ↑ Generation time/age = ↑ Exposure
Development stages = ↑ or ↓ exposure
↑ Contact to contaminated environment = ↑
Exposure
Surface uptake ↑ surface-to-volume ratio = ↑ Exposure
↑ Permeability = ↑ uptake
↑ Discrimination against contaminated food
= ↓ Exposure
Dietary intake Diet
Feeding behaviour
Digestive uptake ↑ complex digestive system = ↑ Cd uptake
↓ pH of the digestive system = ↑ Cd uptake
Regulation of uptake ↑ demand for Zn and Cu = ↑ Cd uptake
↑ regulation = ↓ Cd uptake
Long lived organisms have long exposure time.
Habitat and diet can vary in different development stages. See habitat preferences and diet.
In habitats with close contact to the contaminated environment, e.g. soil or litter phase, exposure is higher
than in habitats with less direct contact to the contaminated environment, e.g. the soil surface.
High surface-to-volume ratio results in a relatively larger exposure to the contaminant. Most important for
permeable organisms.
Soil dwelling organisms, particularly soft-bodied organisms, can take up heavy metals either actively or by
diffusion.
Active rejection of contaminated food, e.g. via chemoreceptor sensoring, can decrease the exposure.
Different types of diet can contain different Cd concentrations, e.g. prey organisms can have up or down
concentrated Cd, compared to organic matter.
Cd concentration can vary within the different parts of a prey, feeding on specific prey parts can thus result
in varying Cd uptake of the predator.
Complex digestive systems, well developed and efficient nutrient assimilation increases the risk of
concurrent Cd assimilation.
Decreased pH in some parts of the digestive tract can increase Cd availability, by breaking the ingested Cd-
bound complexes, e.g. organic matter for detrivores or tissue for predators.
Due to its similarity with Zn and Cu (see Cd toxicity), Cd can be taken up along the same pathways as Zn
and Cu. When Zn and Cu are in demand, the risk of taking up Cd increases.
It is still uncertain if Cd is regulated in some species. If there is a regulating mechanism, Cd can potentially
be excluded from uptake.

4.2.2. Metal detoxification, storage and biomagnification
Where bioaccumulation is defined as the concentration increase of a
substance relative to the environmental concentration through any kind
of uptake process, biomagnification is defined as the concentration
increase within an organism, compared to the concentration in its food
(Janssen et al., 1993). Biomagnification thus depends on the consumer's
ability to take up the substance in question from the food source, and its
ability to store and/or eliminate the substance.
Detoxification of Cd has been intensely studied across a range of
taxonomic groups (Nordberg, 2009; Rainbow, 2002; Wallace and
Lopez, 1997), but studies on terrestrial organisms tend to focus on
isopods, oligochaetes and gastropods (Vijver et al., 2004). The metal is
inactivated, and thus detoxified, by binding to proteins and/or by the
formation of inclusion bodies (Cherian and Nordberg, 1983; Tyler et al.,
1989; Vijver et al., 2004). The metal is thus sequestered away from
molecular sites of toxic action. Detoxification enhances the survival
probability and thus the probability of metal transfer to higher trophic
levels via predation.
The intracellular metallothionein (MT) proteins are particularly
important for Cd detoxification across almost all taxa (Cherian and
Nordberg, 1983; Klaassen et al., 1999, 2009; Stürzenbaum et al., 2004).
MTs are low molecular weight, cysteine-rich, metal-binding proteins
that occur throughout the animal kingdom, in eukaryotic microorgan-
isms and some prokaryotes (Newman, 2003; Vijver et al., 2004). MT is
quickly synthesized in response to Cd exposure and is usually the first
scavenger mechanism (Vijver et al., 2004) and detoxifies Cd efficiently
(Stürzenbaum et al., 2004; Vijver et al., 2006a). The second step in
detoxification is the formation of inclusion bodies, also termed metal-
rich granules (MRG), in the cytoplasm. It is assumed that the inclusion
bodies are composed of membrane enclosed MTs (Hopkin, 1989). MRGs
are usually stored either in the cells (e.g. in the lysosomal system),
diffusely in the tissues (e.g. chloragogen tissue, mineral deposits), or in
particular organs (e.g in the hepatopancreas or in the gut) (Tyler et al.,
1989; Vijver et al., 2004; Wallace et al., 1998). For most terrestrial
organisms, the digestive system is the end storage point. Where MT
protects against short-term exposure, MRG is involved in protection
against long-term exposure (Vijver et al., 2004). Once formed, the MRG
does not appear to be broken down intracellularly (Hopkin, 1989), and
thus must be excreted to avoid bioaccumulation. See Table 3 for sum-
mary of detoxification steps.
Chloragocytes are the most investigated metal sequestration me-
chanism in earthworms (Hopkin, 1989). They compose the
chloragogenous tissue, which lines the gut wall (Andersen and Laursen,
1982a) and consists of MRGs. Earthworms do not excrete Cd, but se-
quester the metal permanently in the gut lining (Andersen and Laursen,
1982a). Release only occurs after death and subsequent lysis of cells
(Vijver et al., 2004). The chloragogenous tissue is the main storage
point for Cd, and earthworms thus have a high accumulation potential
(Andersen and Laursen, 1982a; Morgan et al., 1999; Stürzenbaum et al.,
2004).
The main metal storage organ for arthropods and molluscs is the
hepatopancreas. Although the hepatopancreas only constitutes 5% of
the isopod, it can store 95% of the whole Cd body burden, but when the
hepatopancreas is saturated, Cd may “spill over” and deposit in the
hindgut (Hopkin, 1989). Due to differences in storage cells in the he-
patopancreas, Cd has a longer retention time than other heavy metals
(Hopkin, 1989; Hames and Hopkin, 1991).
Microorganisms are able to detoxify and accumulate relatively large
quantities of heavy metals (Gall et al., 2015). Microorganisms can de-
toxify intracellular heavy metals via binding to MTs or inclusion bodies,
such as polyphosphate granules (Gadd, 1990). Furthermore, fungi can
store and accumulate Cd in vacuoles (Gadd, 1993). Microorganisms can
also bind heavy metals to the cell wall, in order to avoid intracellular
uptake (Gadd, 1990).
Immobilized metals can either be stored or excreted. When metals
are stored in the organism, the potential for bioaccumulation of the
metal in the food web increases (Vijver et al., 2004). Whereas earth-
worms and isopods accumulate Cd, collembolans store Cd in the gut
epithelium where it is excreted by intestinal exfoliation during
moulting (Van Straalen et al., 1987). The Cd concentration in col-
lembolans therefore fluctuates periodically. Excretion of Cd is the most
important parameter for metal concentration within an organism
(Janssen et al., 1991; Janssen and Hogervorst, 1993). However, Cd
storage efficiency has the greatest effect on biomagnification (Janssen
et al., 1991).
The physiochemical form in which Cd is stored in the prey affects
the possibilities for biomagnification (Rainbow, 2002). Cd bound in
soluble complexes, such as MT, organelles and other proteins, are more
likely transferred from the prey to the predator than Cd bound in in-
soluble complexes, e.g. MRG and cellular debris (Vijver et al., 2004;
Wallace and Lopez, 1997; William and Samuel, 2003). The insoluble Cd
is directly excreted, whereas soluble Cd-complexes are taken up and
incorporated into the predator. For invertebrates, most research on the
soluble fractions is performed on aquatic species, however the

456
L.H. Mortensen et al. Ecotoxicology and Environmental Safety 156 (2018) 452–462

Table 3
Summary of detoxification, storage and elimination mechanisms influencing bioaccumulation of cadmium.
Mechanisms How and where Elimination

1st action – detoxification
2nd action – detoxification/
storage
3rd action – storage
Metallothionein (MT)
Inclusion bodies/metal rich
granules (MRG)
Storage organs/sites
Inside the cell, Cd binds to the thiol groups of the MT, which inhibits Cd If Cd is not taken up and bound to MT
interaction with vital functions in the cell, thus effectively detoxifying it. it can be eliminated through the
faeces.
Cd/Cd-MT collects in the cell and is enclosed by membrane, before If Cd is taken up, bound to MT but not
transport to a storage organ/site. stored it can be eliminated through the
faeces.
Storage site for terrestrial organisms is most often in different forms/cells Stored Cd can be eliminated via
of the digestive system. E.g., for arthropods and molluscs it is usually in intestinal exfoliation during moulting.
the hepatopancreas, which is part of the midgut or gut epithelium. For
earthworms it is usually in the chloragogen tissue which is lining the gut.

mechanisms seem to be the same across invertebrates (William and For example, because of the high permeability of the epidermis,
Samuel, 2003). For instance, metals bound in granules in the hepato- Oligochaeta consistently show high Cd accumulation potentials. Also,
pancreas of woodlice (Porcellio scaber), were not transferred to the related species are more likely to have the same detoxification and
predatory spider Dysdera crocata, but metals bound to the iron-binding storage mechanisms (Crommentuijn et al., 1994) which are the most
protein ferritin were (Hopkin, 1989). determining factor for bioaccumulation (Janssen et al., 1993; Wang and
In conclusion, Cd biomagnification requires that it is stored in Fisher, 1999; William and Samuel, 2003). Probably, organisms which
bioavailable form and further that Cd containing tissues of the prey are show no detrimental effects while containing a relatively high Cd
actually consumed by the predator. Table 3 summarizes detoxification, concentration have a well-developed detoxification and storage me-
storage and elimination mechanisms influencing Cd bioaccumulation. chanism. These are the most important species/species groups for bio-
magnification. For detoxification and storage data on specific species
and species groups, see Table S1, which lists detoxification mechanism,
4.3. Conclusions on bioaccumulation storage site and excretion rate for individual soil species, as well as a
summary for species groups.
Predicting the risk of bioaccumulation of Cd in a food web solely Several rankings of total Cd concentration and thus detoxification
based on Cd concentration in the soil, organisms present and the ability in different soil invertebrates show the same general pattern
number of trophic levels is a very complex task. However, key factors (Heikens et al., 2001; Janssen and Hogervorst, 1993; Janssen et al.,
and organism groups can be identified in order to facilitate an esti- 1993): Earthworms ≥ isopods ≥ gastropods ≥ pseudoscorpions ≥
mation of Cd accumulation risk. First, the potential for bioaccumulation diplurans ≥ gamasid mites > spiders ≥ harvestmen ≥ oribatid
is highly dependent on the Cd-soil interaction: The most important mites > diplopods ≥ collembolans > millipedes ≥ carabids ≥ sta-
factor for Cd bioavailability is soil pH followed by organic matter phylinids ≥ curculionids ≥ ants. Hence, food webs with earthworms,
content. As pH and organic matter increase Cd is immobilized and isopods and/or gastropods are of greatest risk of Cd bioaccumulation to
becomes less likely to bioaccumulate. Knowledge of the concentration higher trophic levels.
of bioavailable Cd in the soil, can give some indication of the risk of Cd Most studies on heavy metal accumulation in soil organisms focus
bioaccumulation. Many studies use the soluble faction of Cd as the soil on earthworms (Vijver et al., 2004). Specific studies on Cd bioaccu-
background concentration (Anderson and Christensen, 1988; Blume mulation mechanisms for other organisms are scattered and incon-
and Brümmer, 1991; Brown et al., 2004; Christensen, 1989; Janssen sistent. However, based on studies of specific species uptake, storage
et al., 1997; Kim et al., 2015; Lee et al., 1996; Paradelo et al., 2011; and excretion factors we have estimated the risk of bioaccumulation for
Sauvé et al., 2000a), but insoluble Cd, bound to soil particles, can also other organism groups. As seen in Table 4 and Table S1 our findings
be taken up by some soil organisms and potentially bioaccumulate
(Notten et al., 2005; Scheifler et al., 2003). The physicochemical con-
Table 4
ditions may change from the soil to the digestive system, which can Summary of estimates of the risk of Cd bioaccumulation and biomagnification
reduce the strength of Cd-binding to particles and therefore increase Cd for terrestrial organisms based on detailed information presented in Table S1.
uptake over the cell membrane (Lanno et al., 2004). Hence, for soil Estimates on accumulation risk are based on excretion efficiencies. Estimates on
systems it is important also to consider the insoluble Cd fraction. Metal biomagnification risk are based on storage site and storage form combined with
speciation (e.g. the soluble Cd forms, such as Cd++ or CdCl+ and Cd- Vijver et al.’s (2004) hypothesis of the importance of internal metal fractions for
humate) may give a more realistic measure of background concentra- trophic transfer. Bold signifies key species groups important to consider before
tion, or even Cd concentration in the primary source of the food web, (and after) ash amendment.
e.g. in plants and/or prey. However, in practice, it can be very chal- Cd accumulation Risk of biomagnification
lenging to determine Cd concentration of the primary source in the risk to higher trophic levels
field, because both grazers and predators usually consume different
Oligochaeta Earthworms High High
food items. It thus follows that when evaluating Cd accumulation after
Enchytraeids High High
ash amendment, Cd measuring should be considered carefully, in order Isopoda High Medium
to get the most realistic result. Gastropoda High High
Second, the specific interactions between Cd and the organisms in Collembola Low Low
Arachnida Spiders High Medium
questions, such as exposure intensities and detoxification/storage ca-
Mites Very species Very species dependent
pacities, must be taken in to account. Soil and litter dwelling organisms dependent
are often in direct contact with the contaminant and are thus highly Nematoda Too few studies to Too few studies to draw
exposed. Because of differences in life histories, results for Cd exposure draw conclusions conclusions
on specific species are not necessarily applicable for other species, not Microorganisms Protists Too few studies to Too few studies to draw
draw conclusions conclusions
even if they have the same diet, are on the same trophic level or are
Fungi High Medium
taxonomically closely related. However, closely related species may Bacteria Low Low
show some accumulation similarities, due to shared evolutionary traits.

457
L.H. Mortensen et al.
correlate well with the study of Janssen et al. (1993) on Cd con-
centration in organisms. We found that high Cd accumulators are the
groups Oligochaeta, Isopoda and Gastropoda. We recommend density
estimation of these 3 groups before ash application, in order to estimate
the risk of Cd accumulation. We also recommend monitoring of Cd
content of these 3 groups after ash amendment.
5. Risk of Cd bioaccumulation in ash-amended soils
Due to the risk of toxic effects and trophic transfer of Cd, the Cd
concentration in ash from biofuel is usually the restricting factor for ash
amendment to soil (Pitman, 2006). Bioavailability and accumulation of
Cd after ash application depend on the chemical composition and
physical form of the ash, as well as the physicochemical properties of
the soil and the organisms present in the system (Karltun et al., 2008;
Omil et al., 2007). When amending ash to a soil, we cannot control soil-
and biological factors, however, we can to some extent control the
impact of ash amendment by choosing an ash based on its physio-
chemical properties.
5.1. Importance of ash composition for bioaccumulation
The composition and alkalinity of ash from biofuel varies con-
siderably. The quality depends on the type of fuel/wood, the combus-
tion temperatures (Etiégni and Campbell, 1991), type of incinerator and
storage conditions (Demeyer et al., 2001; Karltun et al., 2008). The Cd
concentration in ash can vary considerably depending on the above –
from 1 to 58 mg Cd kg−1 (Korpilahti et al., 1998; Skov and Ingerslev,
2011). Hence, wood ash application may contribute significantly to the
soil Cd pool. However, governmental authorities set trigger values for
ashes used for soil amendment in order to minimize Cd in the soil, e.g.
20 and 30 g Cd kg−1 ash in Denmark and Sweden, respectively (Danish
Ministry of the Environment, 2008; Swedish National Board of Forestry,
2002).
After incineration, the ash reacts with oxygen, which forms a variety
of oxides. In subsequent contact with water, hydroxides are formed and
the ash becomes highly alkaline (Steenari et al., 1999). Prior to com-
mercial use of ash, it is usually necessary to stabilize the ash by self-
hardening or granulation. During self-hardening or granulation, the ash
is added water in order to form larger and denser particles. This de-
creases the solubility of the ions in the ash, which minimizes a rapid
increase in pH and leaching of nutrients (Steenari et al., 1999). As pH is
the main factor for Cd availability, an obvious assumption would be
that Cd is more bioavailable after application of granulated ash than
loose ash. However, granulated ash also binds the elements in tighter
complexes, and Cd is thus not as easily released from granulated as from
loose ash (Kepanen et al., 2005).
Studies started before the 1990's were usually performed with loose
ash, whereas amendments in more recent times use granulated or self-
hardened ash (Omil et al., 2013). Thus, long-term studies (> 10–20
years after ash application) have used loose ash, the composition of
which rarely has been described; hence cation exchange capacity, al-
kalinity and sometimes element composition are unavailable today
(Moilanen et al., 2002; Saarsalmi et al., 2012). Because of this gap in
the basic knowledge on the ash used in old experiments, the few long-
term studies that exist today can be hard to use for predicting the long-
term consequences of ash application. Generally, the physicochemical
differences between ashes complicate comparisons of ash studies.
However, if future studies state element composition, cation exchange
capacity and pH of the ash in question, it will at least give grounds for
informed comparisons.
5.2. Ash effects on soil properties affecting bioaccumulation potential
The main consequence of applying ash to a system is an increase in
soil pH, especially in acidic soils. Hence, addition of ash and subsequent
458
Ecotoxicology and Environmental Safety 156 (2018) 452–462
pH increase decreases Cd availability (Kepanen et al., 2005; Maresca
et al., 2017). However, because of the complex properties of ash, it can
be difficult to separate the pH effect on Cd availability from other
properties of ash, as well as soil properties. Ca is also a major compo-
nent in ash and significantly increased Ca concentration has been found
up to 16 years after ash application (Saarsalmi et al., 2001). Ca in ash
could potentially increase Cd availability and maybe even counteract
the pH effect. However, modelling pH, Ca concentration and Cd solu-
bility from a liming experiment, Oste et al. (2001) found that pH was
the principal factor for Cd availability. Indeed Ca-rich clay has suc-
cessfully been used to remedy contaminated sediments (Yin and Zhu,
2016).
High Cl concentrations also increase Cd availability; however, ash
does not contain Cl (e.g. (Demeyer et al., 2001)) and thus only influ-
ences Cd availability, if ash is applied to a Cl-rich soil.
High organic matter content of the soil reduces Cd availability. Ash
application has shown not to affect soil organic matter content
(Saarsalmi et al., 2001) hence, organic matter as a factor for Cd bioa-
vailability depends on the soil to which the ash is applied, e.g. peat
compared to mineral soil (Kepanen et al., 2005).
Because ash and soil are composed of complex material with many,
often varying elements, not one parameter for ash or soil can predict the
availability of Cd. However, several studies show that even though ash
application increases the total Cd concentration of the soil, Cd avail-
ability does not increase, not even for ash spiked with Cd (Perkiömäki
al., 2003). Often Cd availability even decreases (Hansen et al., 2001;
Kepanen et al., 2005; Lodenius, 2003; Moilanen et al., 2006), as seen for
the grass Deschampsia flexuosa, where Cd concentration in the roots
decreased with increasing ash concentration (Kindtler and Johansen, in
preparation).
Despite the insignificant change in Cd uptake after application, the
soil holds a larger Cd reserve after ash amendment. With time, soil
alkalinity will decrease and when it reaches a certain threshold (de-
pending on e.g. organic matter of the soil), the Cd reserve may become
available for uptake. A recent study has modelled long-term leaching of
elements from ash, where the cumulative release of Cd was less than 5%
of the solid content (Maresca et al., 2018). Long-term field studies
specifically on Cd availability after ash application are absent (Huotari
et al., 2015), but studies which have investigated pH, Ca concentration
and Cd concentration after several years (Bramryd and Fransman,
1995; Moilanen et al., 2006; Saarsalmi et al., 2001, 2010, 2012), cor-
relate with the modelling study; it appears that leaching of elements to
lower soil profiles is very slow, occurring after a considerable amount of
time (> 10 years). This also corresponds with results showing long-
lasting pH effects after ash application (Huotari et al., 2015; Karltun
et al., 2008). 10–19 years after ash application, pH is still elevated
1.4–2 pH units (Karltun et al., 2008) and after 30 years, pH is still
significantly higher than the control (0.1 pH units) (Saarsalmi et al.,
2012). However, with such a minor pH increase, part of the increased
Cd pool could become bioavailable at any time.
5.3. Ash effects on ecosystems affecting bioaccumulation potential
The factors that determine Cd accumulation are uptake-, storage-
and elimination abilities for an organism. Ash application does not af-
fect these species-specific abilities. However, ash application can in-
fluence the composition of elements that are available for uptake,
abundance of organisms and hence species composition in the com-
munity, which in turn can influence Cd accumulation at the ecosystem
level.
Ash is rich in a variety of ions, but specifically for Cd accumulation,
the Zn content is important. Uptake of Cd and Zn into organisms have
been shown to correlate in heavy metal polluted areas (Janssen and
Hogervorst, 1993; William and Samuel, 2003), which suggest a similar
mode of uptake for the two elements. A similar mechanism for uptake
can result in uptake “competition” between Cd and Zn. In a Cd-Zn
L.H. Mortensen et al.
interaction experiment with the earthworm Aporrectodea caliginosa, a
small addition of Zn to the Cd treatment significantly decreased the
mortality of A. caliginosa, compared to Cd-only treatment (Qiu et al.,
2011). Zn acted as a “protector” against Cd toxicity, probably by
competing for uptake sites. With ash application varying amounts of Cd
are added to the soil, but Zn is simultaneously applied at 33–265×
higher concentrations (Demeyer et al., 2001). Given that Cd and Zn
uptake follow the same pathway, it is therefore 33–265 times more
likely that Zn rather than Cd is taken up.
Some organisms accumulate Cd to a higher degree than others (e.g.
earthworms, isopods, gastropods and mycorrhizae – see Table 4 and
Table S1). If ash application enhances the abundance of organisms with
high Cd accumulation potential, they are more likely to be encountered
as prey and transfer Cd to higher trophic levels. Correspondingly, if
organisms with low Cd accumulation potential (e.g. collembolans) in-
crease, the risk of Cd transfer decreases. Changes in abundance and
biomass of soil fauna after ash application are most likely caused by the
pH change of the environment (Haimi et al., 2000; Qin et al., 2017).
Because the pH effect depends on both ash dose and soil conditions, its
magnitude, and hence impact on organism communities, varies. Ash
effects on abundance and biomass of soil fauna vary both between and
within organism groups (e.g. Nieminen, 2011). For example, effects on
enchytraeids have both shown a decrease (Haimi et al., 2000; Huhta
et al., 1986; Liiri et al., 2001) and an increase in biomass (Qin et al.,
2017). The ambiguous results are probably due to different test species;
Cognettia sphagnetorum and Enchytraeus crypticus, respectively. C.
sphagnetorum has a very narrow pH range, where E. crypticus has a
wider pH range centred around neutral (Jänsch et al., 2005).
Generally, earthworm abundances are low at acidic conditions and
highest at near-neutral soil pH, and it is therefore not surprising that
earthworm abundances increase with ash addition (Huhta et al., 1986,
Mortensen et al., unpublished). Given that Cd accumulation and bio-
magnification are generally high for earthworms, the risk of Cd
bioaccumulation is highest in systems with high earthworm abundances
or in systems, where ash application notably enhances earthworm
abundances. It can thus be an advantage to apply granulated ash, which
will increase pH slower and delay a change in community structure.
Isopods and gastropods also accumulate Cd, but to our knowledge there
are no reports on the impact of ash application on isopod and gastropod
abundances. Many fungal species, including mycorrhizal fungi, accu-
mulate Cd. In most cases ash application left total fungal biomass un-
altered (Liiri et al., 2002; Perkiömäki and Fritze, 2002,
Perkiömäkiet al., 2003), but in a pot experiment ash slightly enhanced
fungal biomass (Mahmood et al., 2003), whereas mycorrhizal coloni-
zation or abundance was either stimulated by (Cruz-Paredes et al.,
2017; Hagerberg and Wallander, 2002; Majdi et al., 2008) or un-
responsive (Taylor and Finlay, 2003) to ash application.
Ash application stimulates microbial organic matter decomposition
(Moilanen et al., 2002; Perkiömäki et al., 2004). A reduction in the
organic matter content could potentially increase Cd bioavailability.
However, despite increased decomposition rates, ash does not seem to
lead to detectable changes in organic matter content (Saarsalmi et al.,
2001).
Cd in ash has so far not shown any harmful effects on micro-
organisms (Perkiömäki and Fritze, 2005), even after ash spiked with Cd
(Perkiömäki and Fritze, 2003; Perkiömäkiet al., 2003). Studies on the in
situ effect of Cd in ash on soil invertebrates are scarce, but no obvious
toxic effects have been observed (Lodenius et al., 2009; Lundkvist,
1998). In spite of this, Cd could become a concern; the soil will hold a
Cd reserve, which potentially could become available in the long term.
However, at biofuel production or other clear cutting sites, the natural
Cd concentration in the system is reduced with removal of biomass.
6. Conclusion and synthesis
Ash from biofuel production is a useful product for soil amendment
459
Ecotoxicology and Environmental Safety 156 (2018) 452–462
to counteract acidification or nutrient depletion. It is possible to esti-
mate the risk of Cd accumulation after ash application by carefully
considering the two variables: ash and soil system. In practice, the
composition of ash is much easier to control than the soil chemistry and
organism community. The ash can thus be selected based on certain
criteria, where a soil system can be evaluated for Cd bioaccumulation
risk.
We here list the key parameters to study and our recommendations
for ash amendment to soils:
1) Ash
a. Cd concentration: High Cd concentration in the ash will lead to a
high Cd reserve in the soil. The threshold values for Cd in ash
used for amendment is 20 and 30 µg Cd g−1 for Denmark and
Sweden, respectively (Danish Ministry of the Environment, 2008;
Swedish National Board of Forestry, 2002), but the Cd con-
centration can be as low as 1 µg Cd g−1 for wood ash (Korpilahti
et al., 1998). A study has shown that even ashes below the trigger
value (16.3 µg Cd g−1 ash), can relase quantities of Cd which are
close to the Danish soil quality criteria (Maresca et al., 2017) of
0.5 µg g−1 (Danish Ministry of the Environment, 2015).
Thus, ashes with low Cd concentration are recommended.
b. Alkalinity: High alkalinity of the ash will cause a greater pH in-
crease in the soil and thus decrease the solubility of Cd and other
elements. However, a greater pH increase will also cause a larger
impact on the soil organism community. The pH of ashes can vary
between 8.9 and 13.5 (Demeyer et al., 2001).
Thus, in order to achieve the desired alkalinity of the soil, but
minimize rapid changes in the organism community, ashes with a
moderate alkalinity are recommended.
c. Post treatment: Loose ash (not post treated) will cause a more
rapid pH increase, Cd and nutrient release from ash particles,
than treated ash. Self-hardened or granulated ash increases pH
slower and binds Cd and other elements tighter.
Thus, in order to minimize a rapid change in pH- and organism
community, granulated ash is recommended.
2) Soil system
a. pH of soil: pH is the most important factor controlling Cd avail-
ability; low pH increases Cd availability. However, ash is most
often used to counteract acidification. It hence follows that a soil
chosen for ash amendment is probably acidic.
The key challenge is to match soil and ash pH in order to achieve
a suitable pH increase in the soil. At pH 3, 75% of the soil Cd is
available (Cd++); at pH 6 only 35% is available. However, at pH
8, the available pool has increased to 55% (Sauvé et al., 2000b).
Thus, pH around 6 would be a suitable end point for a soil, in
relation to Cd availability. It must be kept in mind that any in-
crease in pH will influence the soil organisms.
b. Organic matter content: Organic matter is the second most im-
portant factor controlling Cd availability. High organic matter
content can, also in the long term if/when the pH effect subsides,
reduce Cd availability. Furthermore, organic matter will also
improve ash as a fertilizer, due to possible mineralization and
thus availability of N.
Thus, soils with high organic matter content are recommended.
However, two issues will have to be noted: 1) Ash amendment of
Danish soils is restricted by the existing Cd
concentration: < 0.5 mg Cd kg−1 DW soil (Danish Ministry of the
Environment, 2008). This trigger value does not consider the
volume weight of organic vs mineral soil. Soils high in organic
matter, will thus reach the trigger value quicker than mineral
soils, with the same ash dose. 2) The pH change can enhance
decomposition of soil organic matter. This can potentially change
the net system C balance and increase N mineralization. En-
hanced N mineralization at increased soil pH can increase ni-
trification and denitrification rates and thereby the risk of nitrate
L.H. Mortensen et al.
leaching and emission of greenhouse gases such as N2O.
c. Organism community. If the organisms present are high Cd ac-
cumulators, e.g. earthworms, the risk of accumulation will in-
crease, especially in cases where ash application increases the
abundance of high Cd accumulators. However, if the organisms
present are negatively affected by the changes in pH, their
abundance will probably decline and the risk of bioaccumulation
can decrease. Other species may occupy the free niche after a pH
increase, and bioaccumulation depends on the Cd accumulation
potential of these species. Here it should be noted that earth-
worms will often be stimulated by increased pH following wood
ash addition to acidic soils (Huhta, 1986, Mortensen et al., un-
published)
An analysis of the organism groups present in the soil system is
recommended in order to estimate risk of Cd accumulation. Or-
ganisms of interest can be viewed in Table 4 and Table S1, which
summarize risk of Cd transfer to higher trophic levels for a range
of soil organisms.
It is always difficult to predict heavy metal accumulation in a given
system, but with a few well-chosen tests on the targeted soil system and
some careful considerations it is possible to minimize Cd accumulation:
The chosen ash should be granulated with low Cd concentration and
moderate alkalinity. The intended soil should be tested for pH and low
Cd content, high organic matter content and low density of the key
organisms: oligochaetes, isopods, gastropods.
Acknowledgements
This work was supported by the “Center for Bioenergy Recycling –
ASHBACK” funded by the Danish Council for Strategic Research (Grant
no. 0606-00587B). We appreciate the constructive comments of two
anonymous reviewers, which significantly improved the manuscript.
Appendix A. Supplementary material
Supplementary data associated with this article can be found in the
online version at http://dx.doi.org/10.1016/j.ecoenv.2018.03.018.
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