Science of the Total Environment 628–629 (2018) 556–561

Contents lists available at ScienceDirect

Science of the Total Environment

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

Cigarette butts may have low toxicity to soil-dwelling invertebrates:

Evidence from a land snail
Hussan Gill, Kyler Rogers, Bilal Rehman, John Moynihan, Elizabeth A. Bergey ⁎
Oklahoma Biological Survey and Department of Biology, University of Oklahoma, Norman, OK 73019, USA

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Cigarette butts are commonly littered

on the ground.
• Toxicity of cigarette butts to soil-associ-
ated organisms was unexplored.
• Cigarette butt effluent did not impact
land snail survival, growth or feeding.
• Snails avoided fresh whole cigarette
butts but avoidance subsided within a
month.
• Cigarette butts may have low toxicity to
soil-dwelling invertebrates.

a r t i c l e i n f o a b s t r a c t

Article history: Cigarette butts are a common form of litter that is often deposited on soil, where toxins from butts may affect soil-
Received 18 December 2017 dwelling organisms. We examined possible toxicity of cigarette butts to the woodland snail Anguispira alternata
Received in revised form 4 February 2018 using a toxicity study with cigarette butt effluent and a lab-based habitat choice experiment in which snails could
Accepted 7 February 2018
feed or rest on areas with different butt densities. No mortality occurred during the 32-day toxicity study, which
Available online xxxx
used six effluent concentrations ranging from 0 to 4 butts/l (0 to 0.92 butts/kg of soil). Neither food consumption
Editor: Jay Gan nor snail growth differed among the effluent concentrations. When provided a choice among four habitats with 0
to 4 cigarette butts, snails selected to preferentially rest in the 0-butt habitat and avoided the 4-butt habitat. This
Keywords: distribution pattern was strong during the first wk. but became weaker over time and largely disappeared by the
CBs end of the 3-wk experiment. Snails did not discriminate among butt densities when feeding. This is the first tox-
Cigarette butt leachate icity test using cigarette butts on soil-dwelling invertebrates. Declining aversion to cigarette butts over a 3-wk
Gastropods period may indicate declining toxicity of terrestrially deposited butts as they age, but further testing is needed.
Aversion © 2018 Elsevier B.V. All rights reserved.

1. Introduction
Cigarette smoking produces chemically complex by-products. The
toxicity of cigarette smoke, both directly to smokers and indirectly
through second-hand smoke has been well documented and there is
⁎ Corresponding author at: 111 E. Chesapeake Street, Oklahoma Biological Survey and
Department of Biology, University of Oklahoma, Norman, OK 73019, USA.
E-mail address: lbergey@ou.edu (E.A. Bergey).
increasing concern with thirdhand smoke, the residues left after
smoke dissipates (Matt et al., 2011). A fourth component, cigarette
butts, are also toxic (e.g., Booth et al., 2015; Lee and Lee, 2015;
Slaughter et al., 2011) but this toxicity has received less study. Cigarette
butts form a major litter type in urban areas (Bator et al., 2011;
Moriwaki et al., 2009; Roder Green et al., 2014) and, because they can
be moved by surface runoff into waterways, are also a litter problem
along shorelines of lakes and oceans (Novotny et al., 2009; Register,
2000).

https://doi.org/10.1016/j.scitotenv.2018.02.080
0048-9697/© 2018 Elsevier B.V. All rights reserved.
 The toxicity of these butts is a newly recognized and growing area of
interest, in part because of the high number of littered butts. Both the fil-
ters and unsmoked tobacco in cigarette butts contain a wide variety of
chemicals (Moriwaki et al., 2009; Pelit et al., 2013; Shevchenko, 2012)
rendering butts toxic (reviewed by Novotny and Slaughter, 2014).
Worldwide, about 6.25 trillion cigarettes are smoked per year (2012 es-
timate; Ng et al., 2014) and although smoking prevalence is decreasing,
population growth has resulted in increasing cigarette consumption be-
tween 2008 and 2012 (Ng et al., 2014). Much of the resulting mass of
cigarette butts is deposited in the environment. Smokers often litter cig-
arette butts, in part because some smokers do not consider environ-
mentally discarded butts to be litter (Bator et al., 2011; Patel et al.,
2013; Rath et al., 2012). The cellulose acetate basis of filters is highly re-
sistant to biodegradation and so much of the butt persists (Bonanomi et
al., 2015; Puls et al., 2011) - and may continue to leach chemicals. In-
deed, cigarette butts are considered hazardous waste (Barnes, 2011).
Laboratory toxicity studies have demonstrated that cigarette butts,
in the form of aqueous leachate, are toxic to aquatic organisms; includ-
ing microorganisms (Vibrio fischeri; Micevska et al., 2006), zooplankton
(Daphnia magna and Ceriodaphnia dubia; Micevska et al., 2006; Register,
2000; respectively), mosquito larvae (Aedes albopictus; Dieng et al.,
2011), tidepool snails (three species; Booth et al., 2015), frogs (Xenopus
laevis larvae; Parker and Rayburn, 2017) and fishes [freshwater flathead
minnows, Pimephales promelas and marine topsmelt, Atherinops affinis
(Slaughter et al., 2011), and embryos of the Japanese rice fish Oryzias
latipes (Lee and Lee, 2015)]. Acute toxicity occurs at levels as low as
one cigarette butt per liter of water.
Despite the frequency of butt litter on land, land-based toxicity stud-
ies have been overlooked. In the one exception, house finches
(Carpodacus mexicanus) incorporate shredded cigarette butts into
their nests, which reduced nest-associated ectoparasites (Suárez-
Rodríguez et al., 2013); indeed, house finches apparently adjust filter
use in response to parasite levels (Suárez-Rodríguez and Garcia,
2017). Whether cigarette butts were toxic or repellent to ectoparasites
was not mentioned in these studies. Incorporating cigarette butts in
nests was associated with higher fledgling success but also a greater in-
cidence of abnormalities in the nuclei of red blood cells, an indicator of
toxicity (Suárez-Rodríguez and Macías Garcia, 2014).
Our research objective was to test for possible toxicity of cigarette
butts to soil-associated invertebrates - targeting land snails. Many
butts end up directly on the soil or mixed with leaf litter, where butts
and effluent from butts can affect soil-dwelling invertebrates, including
558 H. Gill et al. / Science of the Total Environment 628–629 (2018) 556–561
tested: 4, 2, 1, 0.5, and 0.25 butts/l, as well as a 0 butt control (plain de-
ionized, distilled water) (Fig. 1A).
During the experiment, pairs of snails were housed in 473 ml
(1 pint) glass canning jars in which the lids were replaced with 1-mm
plastic mesh. Jars were half-filled with formulated soil, using the sand,
peat, clay, and calcium carbonate mixture specified by the US-EPA for
toxicity studies using red worms (US-EPA, 2012). The EPA soil mixture
was modified by reducing the clay portion (from 20% to 13%; which re-
duced the tendency of the soil to stick to the shells of the snails), propor-
tionally increasing the peat, and using 2% calcium carbonate, which
exceeded the amount needed to neutralize soil pH.
Each treatment (i.e., the six effluent concentrations) was replicated
five times. Jars were numbered and treatments were assigned randomly
to jar numbers, so the treatment in each jar was not apparent during the
experiment. Forty-five milliliters of effluent solution was added to the
150 g of dry soil mixture in each jar, producing a damp soil. A pair of
snails was added to each jar. The shell of one snail of each pair as marked
with yellow paint to distinguish the two snails and the shell diameters
of both snails were measured with digital calipers. Jars where placed
into an environmental chamber, set with a 50:50 light:dark schedule
and 85% humidity (to reduce evaporation, as per EPA protocols). All
jars were spritzed with spring water three times during the experiment
when the soil surface had dried. Spring water (Ozarka, sourced from
various Texas springs; with pH = 8.1; conductivity ~45 μS/cm) was
spritzed, rather than using deionized water, to avoid osmotic distur-
bance to snails, as water could directly contact snails.
During the 32-day experiment (26 May–27 June), snails were fed
disks of Romaine lettuce (diameter = 3.55 cm; area = 9.9 cm2) cut
with a circular cookie cutter. We cut disks from the outer dark green
sections of leaves, which is the preferred area for snail grazing. Disks
were replaced every 2–3 days and, upon removal, the area of each leaf
disk was measured using a scanner and Winfolia software (Regent In-
struments, Quebec, Canada). Measurements were converted to lost
leaf area by subtraction from the original area of 9.9 cm2. Lettuce was re-
placed 13 times during the experiment.
Snails were re-counted and re-measured at the conclusion of the ex-
periment and returned to lab colony area for maintenance. Snails were
released at the collection site following a rain.
Data consisted of snail mortality (none, so no analysis was needed),
snail (shell) growth, and lettuce consumption.
Snail growth was assessed by comparing shell diameters. Shell di-
ameters of the two snails in each jar were averaged before analysis.
The initial shell diameter and the change in shell diameter over the ex-
periment were assessed with 1-way ANOVA. Area loss of the lettuce
disks in each jar was calculated as percent loss over the course of the ex-
periment by summing the area lost over the 13 measurements and di-
viding by the potential total leaf area (=13 ∗ 9.9 cm2 or 128.7 cm2).
Percent leaf area loss across butt effluent levels was tested with 1-way
ANOVA. No data transformations were needed in the ANOVA analyses.
Patterns of leaf loss over the experiment were analyzed with linear re-
gression within each treatment. Initial leaf loss was very low (mean
= 3.3 of the 9.9 cm2, versus 3.1 cm2 loss due to evaporation alone – un-
published data); therefore regressions were analyzed starting at 8 days
(the next data collection time), after a period of snail acclimation. Slopes
of regressions were compared to describe patterns of leaf consumption
among treatments. ANOVA analyses in both experiments were per-
formed using SYSTAT.
2.3. Choice experiment
Eight replicate choice test arenas were constructed by gluing corru-
gated plastic dividers into a round plastic tub, which produced four
equal-sized areas (Fig. 1B). Formulated soil was added to the height of
the tops of the dividers (approximately 3 cm depth) and soil was wetted
with bottled spring water (pH = 8.1; conductivity ~45 μS/cm). The
center of tub lids was removed and replaced with plastic mesh for air
circulation.
Treatments were four levels of intact cigarette butts – 0, 1, 2, or 4
butts per compartment. Treatments were randomly assigned to com-
partments within each arena. This experiment used Anguispira collected
from the same backyard as for the first experiment and the snails were
similarly maintained in the lab prior to the experiment. The experiment
was placed in a quiet room in an area with indirect sunlight. To reduce
evaporation, mesh on lids was partly covered with paper. All tubs
were spritzed with spring water when soil surfaces dried.
At the start of the experiment, 2 adult snails were placed in each
compartment (=8 snails per arena). Snails were fed Romaine lettuce,
pre-weighed pieces of which were placed in each compartment. Snails
were located twice a week in the afternoon, their resting positions
noted and lettuce was removed, weighed and replaced with weighed
fresh lettuce. Snails were typically located just below the soil surface.
The experiment lasted 3 weeks (22 September to 13 October).
Data for snail location with respect to cigarette butt density were in
the form of counts of the number of snails in each compartment on each
of the six sampling dates. These count data were analyzed using Chi-
square tests for the total counts of snails in each of the four treatment
compartments (0, 1, 2, and 4 cigarette butts) over the six data-collection
dates. With 8 arenas, each with 8 snails and 4 compartments, and 6
sampling dates, the expected number of snails was 96 snails per com-
partment (8 arenas × 8 snails × 6 dates / 4 compartments). The percent
contributions of each treatment toward the overall Chi-square value
were determined to assess the treatments contributing most to the sig-
nificant Chi-square value. In addition, Chi-square analysis was used to
assess temporal patterns of snail distributions across treatments by
assessing the distribution of snails on each sampling date (expected
values were 16 snails per compartment: 8 arenas × 8 snails / 4 compart-
ments). Chi-square analyses were done ‘by hand’ on an Excel datasheet.
Lettuce consumption in each compartment was assessed as the pro-
portion of lettuce consumed on 5 of the 6 sampling dates. Data from the
first sampling date (day 8) was not included because snails were under-
fed and consumed all the lettuce in most of the compartments. Excess
lettuce was provided during the rest of the experiment. Because lettuce
was weighed and replaced in unequal intervals (twice a week; split into
5 days and 2 days), data were standardized as the proportion consumed
per day. Data failed both normal distribution and equal variance tests
and, as a consequence, were arcsine square-root transformed. These
transformed data were analyzed with a repeated-measures, 2-way
ANOVA. Arenas were the repeated measure and treatments were the
number of cigarette butts in compartments and the day of the experi-
ment. Tukey's test was used to assess significant differences among
treatment levels.
3. Results
3.1. Survival experiment
No deaths occurred among the 60 snails in the toxicity experiment.
There was no significant difference in snail size at the start of the exper-
iment (F5,24 = 1.067; P = 0.403), with snail shell diameter averaging
11.04 ± 0.12 (SE) mm. All snails grew during the experiment and
growth, as increase in shell diameter, did not differ among the six treat-
ments (F5,24 = 0.774; P = 0.578). Final shell diameter averaged 12.14 ±
0.13 (SE) mm.
The percent loss of lettuce area over the experiment was not
significantly different among the cigarette butt treatments (F5,24 =
2.165; P = 0.092). Leaf loss patterns over the experiment were variable
within individual jars and among treatments. Snails initially ate little
and, as a consequence, patterns of consumption were analyzed between
days 8 and 32. Linear regressions at the treatment level showed both
patterns of increasing leaf consumption over the experiment (control,
 H. Gill et al. / Science of the Total Environment 628–629 (2018) 556–561 559

and 0.50 and 2 butts/l treatments) and patterns of decreasing consump-

tion (0.25, 1, and 4 butts/l treatments; Table 1).

3.2. Choice experiment

This experiment separately tested both the choice of resting location

and the location of feeding with respect to cigarette butt density.
Resting snails were distributed unevenly across the different com-
partments within the arenas (Χ2 = 36.77, P b 0.001; Fig. 2A), with a pat-
tern of reduced snail frequency with increasing cigarette butt density. A
high frequency of snails in the control compartment and lower fre-
quently in the four cigarette butt treatment contributed most of the
total Chi-square value (48% and 50%, respectively).
The distribution of snails across the cigarette butt treatments varied
over the 22-day experiment (Fig. 2B). The pattern of high snail fre-
quency in the control compartment and low frequency in the 4-butt
compartment was evident through day eight (P b 0.001). Between
days 8 and 15, this pattern remained but became less pronounced
(0.05 b P b 0.10) and by day 20, snails were no longer choosing the con-
trol (with 0 butts) more frequently than expected. Snails continued to
show a degree of avoidance of the 4-butt treatment, with 10 and 13
snails of an expected 16 snails during the last third of the experiment.
Lettuce consumption did not differ among compartments with dif-
ferent numbers of cigarette butts (P = 0.45; Table 2), nor did the arenas
with different treatments have differences in lettuce consumption. The
day of the experiment factor was significant (P b 0.001); specifically the
daily proportion consumed was greater for feeding intervals of 2 days
versus 5 days (Tukey: P b 0.05). This significant result was considered
an artifact of the experimental design. This species of snail is typically
quiescent, often buried in the soil. Snail activity followed disturbance
during snail counting and activity (and presumably eating) was evi-
dently similar whether data were collected after 2 days or 5 days –
thus daily consumption was higher after 2 days than 5 days. Follow-
up analysis of weight consumed between samplings, regardless of
time, indicated that consumption was higher in the 8–15 day interval
(P = 0.01), but the data were non-normal and normality was not ob- Fig. 2. Distribution of snails among arena compartments during the choice experiment.
tained by transformation. In all analyses, the interaction of time and cig- (A) Sum of snail counts throughout the experiment; (B) Counts on individual sampling
dates. The horizontal dashed lines show the expected Chi-square values [96 snails in (A)
arette butt treatment was not significant.
and 16 snails in (B)]. In (B), ‘**’ = P b 0.001, ‘*’ = 0.05 b P b 0.10, ‘NS’ = Not significant.

4. Discussion
Toxicity testing using cigarette butt effluent in soil showed no toxic
effects on Anguispira survival despite using effluent concentrations up
to 0.92 mg/kg, which is similar to or exceeds concentrations toxic to
tested aquatic organisms (discussed below). The difference in toxicity
between our study and results from aquatic studies has several possible
reasons, including variation among taxa, differences between aquatic
and terrestrial toxicity tests, and possible chemical tolerance in soil-as-
sociated snails. Non-lethal evidence of toxicity by cigarette butt effluent
has been reported (Booth et al., 2015; Lee and Lee, 2015; Slaughter et al.,
2011; Suárez-Rodríguez and Macías Garcia, 2014), but neither snail let-
tuce consumption or growth was affected by cigarette butt effluent
levels in our studies. Variability in lettuce consumption may have ob-
scured any differences in feeding patterns and growth may have been
measured over an insufficient time period to conclude that cigarette
butt effluent has absolutely no effect on these endpoints.
Relatively few taxa have been tested for their toxicity to cigarette
butt effluents and among those that have, species differ in susceptibility.
Unfortunately, comparison among studies is complicated by differences
in methods, including the types of cigarettes and methods of smoking,
period of soaking, and the length of tests. Consequently, within-study
comparisons are most relevant because of methodological similarities.
These studies encompass a range of patterns: similar LC50s in a marine
and a freshwater fish (0.97 and 1.1 butts/l, respectively; Slaughter et al.,
Table 2
Analysis of consumption of Romaine lettuce by snails during the choice test. Lettuce was
provided in all four treatment compartments and the proportion of lettuce lost per day
was tested with a repeated measures, 2-way ANOVA test on transformed data.

Table 1 Source of variation DF MS F P

Linear regression summary of patterns of leaf loss over the experiment (days 8–32). Pos- Arena (repeated measure) 7 0.0177
itive slopes indicate increased feeding over time and negative slopes indicate reduced Cigarette butt treatment 3 0.00331 0.259 0.854 NS
feeding over time. Treatment × RM 21 0.0128
Day of experiment 4 0.121 7.340 b0.001 8a 13b 15a 20b 22a
Treatment Slope Intercept R2
Day of experiment × RM 28 0.0165
Control 0.050 4.77 0.09 Treatment × day of 12 0.0132 1.196 0.299 NS
0.25 butts/l −0.061 7.13 0.11 experiment
0.50 butts/l 0.013 4.46 0.01 Residual 84 0.0110
1.0 butts/l −0.058 7.77 0.07 Total 159 0.0153
2.0 butts/l 0.088 3.58 0.17
NS = not significant; different superscript letters above the day of experiment indicate
4.0 butts/l −0.054 5.73 0.16
significant difference (Tukey test results).
560 H. Gill et al. / Science of the Total Environment 628–629 (2018) 556–561
2011) but differences in EC50 values of planktonic organisms
(0.05 butts/l for immobility in water fleas and 0.7 butts/l for biolumines-
cence in bacteria; Micevska et al., 2006). The closest comparator to our
study is a study comparing toxicity among tidepool-dwelling snails
(Booth et al., 2015), although the terrestrial versus aquatic habitats pro-
vides an added complication. This comparison indicates reduced toxic-
ity in terrestrial versus aquatic snails. Anguispira showed no toxic
response at 0.92 butts/kg after a month, whereas the 50% survival
rates among the three tested species ranged from 1 to 2.75 butts/l
after 1 week (estimated from Fig. 1 in Booth et al., 2015). Studies that di-
rectly compare terrestrial versus aquatic toxicity are rare because few
species occur in both habitats. Some nematodes are an exception and
Boyd and Williams (2003) compared copper toxicity among three spe-
cies of nematodes and the aquatic versus terrestrial average LD50s were
85 versus 179, 19 versus 48, and 160 versus 251 mg Cu/l – a pattern also
showing greater toxicity in an aquatic than in a terrestrial system. Both
the nematode study and our results indicate that toxicity may be greater
to aquatic organisms than to soil dwelling invertebrates, but differences
in environmental attributes and pathways of exposure differ signifi-
cantly between terrestrial and aquatic systems and further research is
needed to identify specific outcomes of cigarette butt leachate
chemicals to land-based species. For example, as a consumer of leaf lit-
ter, Auguispira (and many other land snails) may be tolerant of a variety
of secondary compounds produced by plants, and thus little affected by
the secondary compounds in tobacco and cigarette butts. Specific stud-
ies of the effective toxic levels of cigarette butt effluents to a wider range
of organisms, including land snails, are warranted, as are field measure-
ments of environmental concentrations of cigarette butt effluents or ef-
fluent components in both aquatic and terrestrial habitats.
Land snails have been used to study other toxicants. Snails are often
used in biomonitoring for environmental metals and have occasionally
been used in toxicity testing (reviewed by Cortet et al., 1999). Cornu
aspersum (=Helix aspersa or Cantareus aspersus) has been used in bio-
monitoring or as a sentinel animal because this snail bioaccumulates en-
vironmental metals (Gomot-De Vaufleury, 2000; Gomot de Vaufleury
and Pihan, 2000; Nica et al., 2015; Pauget et al., 2012) and has a high tol-
erance for metals (Cortet et al., 1999; Nica et al., 2015). Other, regionally
appropriate species have been similarly used for metal monitoring in
Europe (Berger and Dallinger, 1993; Boshoff et al., 2013; Emilia et al.,
2016; Fritsch et al., 2011; Madejón et al., 2013; Notten et al., 2006)
and in Japan (Yasoshima and Takano, 2001). In one of the few examples
of toxicity testing using land snails, forestry pesticides were incorpo-
rated into the diet of Cornu, with the result that toxicity varied among
the 12 pesticides (Schuytema et al., 1994). Cornu was also used in a re-
cent study of herbicides, which showed evidence of endocrine disrup-
tion (Druart et al., 2017). Cornu was available for this study, but we
selected Anguispira because this species is native in North America, is
highly soil-associated and typically retracts into it shell when resting,
whereas resting Cornu attach to solid surfaces, especially the sides and
lids of cages in the lab, by sealing its shell to surfaces with dried
mucus – thus avoiding soil-associated toxins.
Although we did not find mortality, Anguispira avoided cigarette
butts, especially dense concentrations of butts. Avoidance is a common
endpoint in toxicology studies (Booth et al., 2015), as are endpoints that
we did not observe – namely, changes in growth and altered food con-
sumption rates (Drobne, 1997). Anguispira were mobile enough to
move among all chambers within arenas and typically burrowed just
beneath the soil surface. Thus, snails were in direct contact with soil
when resting. No locational differences were found in lettuce consump-
tion, perhaps because contact with soil was reduced during feeding be-
cause the snails were on the lettuce.
Avoidance of cigarette butts decreased over time, indicating that the
repellent compounds were volatile, changed chemically or were dis-
persed or diluted. Test chambers initially smelled strongly of cigarette
butts, but the odor lessened with time and was not noticeable after
two weeks (personal observation). Reduction in cigarette butt toxicity
with time was also found by Dieng et al. (2011), in that the ability to
kill or impact mosquito larvae (Aedes albopictus) in a closed system de-
creased over time, but toxic effects resumed with the addition of fresh
cigarette butts. The possibility of changing toxicity over time, especially
of dry butts, has environmental implications. Butts can accumulate over
time and with rain and leaching of butts (especially in puddles) any
resulting runoff may affect aquatic systems - or water may move down-
ward, contaminating groundwater (Roder Green et al., 2014). Toxicity
studies have been done with fresh cigarette butts and our results indi-
cate that terrestrial aging of butts may reduce toxicity, in addition to se-
rial leaching by rain (Roder Green et al., 2014). Most littered butts are
deposited on land and undergo aging before being conveyed to waters,
so further research on aerial emissions (Poppendieck et al., 2016),
chemical composition, and toxicity of aging butts is warranted.
Acknowledgements
Laura Souza generously allowed use of equipment and Jeremy Ross
provided space for experiments. Partial funding was from the University
of Oklahoma's Undergraduate Research Opportunities Program (to HG
and BR).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2018.02.080.
References
Barnes, R.L., 2011. Regulating the disposal of cigarette butts as toxic hazardous waste. Tob.
Control. 20, i45–i48.
Bator, R.J., Bryan, A.D., Schultz, P.W., 2011. Who gives a hoot? Intercept surveys of litterers
and disposers. Environ. Behav. 43, 295–315.
Berger, B., Dallinger, R., 1993. Terrestrial snails as quantitative indicators of environmental
metal pollution. Environ. Monit. Assess. 25, 65–84.
Bonanomi, G., Incerti, G., Cesarano, G., Gaglione, S.A., Lanzotti, V., 2015. Cigarette butt de-
composition and associated chemical changes assessed by 13C CPMAS NMR. PLoS
ONE. Public Libr. Sci. 10 (1), e0117393.
Booth, D.J., Gribben, P., Parkinson, K., 2015. Impact of cigarette butt leachate on tidepool
snails. Mar. Pollut. Bull. 95, 362–364.
Boshoff, M., Jordaens, K., Backeljau, T., Lettens, S., Tack, F., Vandecasteele, B., et al., 2013.
Organ- and species-specific accumulation of metals in two land snail species
(Gastropoda, Pulmonata). Sci. Total Environ. 449, 470–481.
Boyd, W.A., Williams, P.L., 2003. Comparison of the sensitivity of three nematode species
to copper and their utility in aquatic and soil toxicity tests. Environ. Toxicol. Chem. 22,
2768–2774.
Cortet, J., Vauflery, A.G.-D., Poinsot-Balaguer, N., Gomot, L., Texier, C., Cluzeau, D., 1999.
The use of invertebrate soil fauna in monitoring pollutant effects. Eur. J. Soil Biol.
35, 115–134.
Dieng, H., Saifur, R.G.M., Ahmad, A.H., Md Rawi, C.S., Boots, M., Satho, T., et al., 2011.
Discarded cigarette butts attract females and kill the progeny of Aedes albopictus.
J. Am. Mosq. Control Assoc. 27, 263–271.
Drobne, D., 1997. Terrestrial isopods—a good choice for toxicity testing of pollutants in the
terrestrial environment. Environ. Toxicol. Chem. 16, 1159–1164.
Druart, C., Gimbert, F., Scheifler, R., de Vaufleury, A., 2017. A full life-cycle bioassay with
Cantareus aspersus shows reproductive effects of a glyphosate-based herbicide sug-
gesting potential endocrine disruption. Environ. Pollut. 226, 240–249.
Emilia, R., Debora, B., Stefania, A., Nicola, B., Roberto, B., 2016. Papillifera papillaris (O.F.
Müller), a small snail living on stones and monuments, as indicator of metal deposi-
tion and bioavailability in urban environments. Ecol. Indic. 69, 360–367.
Fritsch, C., Coeurdassier, M., Gimbert, F., Crini, N., Scheifler, R., de Vaufleury, A., 2011. In-
vestigations of responses to metal pollution in land snail populations (Cantareus
aspersus and Cepaea nemoralis) from a smelter-impacted area. Ecotoxicology 20,
739–759.
Gomot de Vaufleury, A., Pihan, F., 2000. Growing snails used as sentinels to evaluate ter-
restrial environment contamination by trace elements. Chemosphere 40, 275–284.
Gomot-De Vaufleury, A., 2000. Standardized growth toxicity testing (Cu, Zn, Pb, and pen-
tachlorophenol) with Helix aspersa. Ecotoxicol. Environ. Saf. 46, 41–50.
Horsák, M., Juřičková, L., Kintrová, K., Hájek, O., 2009. Patterns of land snail diversity over
a gradient of habitat degradation: a comparison of three Czech cities. Biodivers.
Conserv. 18, 3453–3466.
Horsák, M., Lososová, Z., Čejka, T., Juřičková, L., Chytrý, M., 2013. Diversity and biotic ho-
mogenization of urban land-snail faunas in relation to habitat types and
macroclimate in 32 central European cities. PLoS ONE 8, e71783.
Hubricht, L., 1985. The distributions of the native land mollusks of the eastern United
States. Fieldiana Zool. 24, 1–191.
 H. Gill et al. / Science of the Total Environment 628–629 (2018) 556–561
Lee, W., Lee, C.C., 2015. Developmental toxicity of cigarette butts – an underdeveloped
issue. Ecotoxicol. Environ. Saf. 113, 362–368.
Lee, J.G.L., Ranney, L.M., Goldstein, A.O., 2013. Cigarette butts near building entrances:
what is the impact of smoke-free college campus policies? Tob. Control. 22, 107–112.
Madejón, P., Arrébola, J., Madejón, E., Burgos, P., López-Garrido, R., Cárcaba, A., et al., 2013.
The snail Theba pisana as an indicator of soil contamination by trace elements: poten-
tial exposure for animals and humans. J. Sci. Food Agric. 93, 2259–2266.
Marah, M., Novotny, T.E., 2011. Geographic patterns of cigarette butt waste in the urban
environment. Tob. Control. 20, i42–i44.
Matt, G.E., Quintana, P.J.E., Destaillats, H., Gundel, L.A., Sleiman, M., Singer, B.C., et al., 2011.
Thirdhand tobacco smoke: emerging evidence and arguments for a multidisciplinary
research agenda. Environ. Health Perspect. 119, 1218–1226.
Micevska, T., Warne, M.S.J., Pablo, F., Patra, R., 2006. Variation in, and causes of, toxicity of
cigarette butts to a cladoceran and microtox. Arch. Environ. Contam. Toxicol. 50,
205–212.
Moerman, J.W., Potts, G.E., 2011. Analysis of metals leached from smoked cigarette litter.
Tob. Control. 20, i30–i35.
Moriwaki, H., Kitajima, S., Katahira, K., 2009. Waste on the roadside, ‘poi-sute’ waste: its
distribution and elution potential of pollutants into environment. Waste Manag. 29,
1192–1197.
Ng, M., Freeman, M.K., Fleming, T.D., Robinson, M., Dwyer-Lindgren, L., Thomson, B., et al.,
2014. Smoking prevalence and cigarette consumption in 187 countries, 1980–2012.
JAMA 311, 183–192.
Nica, D.V., Filimon, M.N., Bordean, D.-M., Harmanescu, M., Draghici, G.A., Dragan, S.,
Gergen, I.I., 2015. Impact of soil cadmium on land snails: a two-stage exposure ap-
proach under semi-field conditions using bioaccumulative and conchological end-
points of exposure. PLoS ONE 13, e0116397.
Notten, M.J.M., Oosthoek, A.J.P., Rozema, J., Aerts, R., 2006. Heavy metal pollution affects
consumption and reproduction of the landsnail Cepaea nemoralis fed on naturally
polluted Urtica dioica leaves. Ecotoxicology 15, 295–304.
Novotny, T.E., Slaughter, E., 2014. Tobacco product waste: an environmental approach to
reduce tobacco consumption. Curr. Environ. Health Rep. 1, 208–216.
Novotny, T.E., Lum, K., Smith, E., Wang, V., Barnes, R., 2009. Cigarettes butts and the case
for an environmental policy on hazardous cigarette waste. Int. J. Environ. Res. Public
Health 6, 1691–1705.
Parker, T.T., Rayburn, J., 2017. A comparison of electronic and traditional cigarette butt
leachate on the development of Xenopus laevis embryos. Toxicol. Rep. 4, 77–82.
Patel, V., Thomson, G.W., Wilson, N., 2013. Cigarette butt littering in city streets: a new
methodology for studying and results. Tob. Control. 22, 59–62.
561
Pauget, B., Gimbert, F., Scheifler, R., Coeurdassier, M., de Vaufleury, A., 2012. Soil parame-
ters are key factors to predict metal bioavailability to snails based on chemical ex-
tractant data. Sci. Total Environ. 431, 413–425.
Pelit, F.O., Demirdöğen, R.E., Henden, E., 2013. Investigation of heavy metal content of
Turkish tobacco leaves, cigarette butt, ash, and smoke. Environ. Monit. Assess. 185,
9471–9479.
Poppendieck, D., Khurshid, S., Emmerich, S., 2016. Measuring airborne emissions from
cigarette butts: literature review and experimental plan. NIST Interagency/internal
Report (NISTIR)-8147.
Puls, J., Wilson, S.A., Hölter, D., 2011. Degradation of cellulose acetate-based materials: a
review. J. Polym. Environ. 19, 152–165.
Rath, J.M., Rubenstein, R.A., Curry, L.E., Shank, S.E., Cartwright, J.C., 2012. Cigarette litter:
smokers' attitudes and behaviors. Int. J. Environ. Res. Public Health 9, 2189–2203.
Register, K., 2000. Cigarette butts as litter-toxic as well as ugly? Underw. Nat. 25, 23–29.
Roder Green, A.L., Putschew, A., Nehls, T., 2014. Littered cigarette butts as a source of nic-
otine in urban waters. J. Hydrol. 519 (Part D), 3466–3474.
Schuytema, G.S., Nebeker, A.V., Griffis, W.L., 1994. Effects of dietary exposure to forest
pesticides on the brown garden snail Helix aspersa Müller. Arch. Environ. Contam.
Toxicol. 26, 23–28.
Seitz, C.M., Strack, R.W., Orsini, M.M., Rosario, C., Haugh, C., Rice, R., et al., 2012. Quantify-
ing littered cigarette butts to measure effectiveness of smoking bans to building pe-
rimeters. J. Am. Coll. Heal. 60, 331–334.
Shevchenko, V., 2012. Characterization of chemical compounds in cigarette filters leach-
ates. Public Health. San Diego State University, San Diego, CA, p. 53 (Masters thesis).
Slaughter, E., Gersberg, R.M., Watanabe, K., Rudolph, J., Stransky, C., Novotny, T.E., 2011.
Toxicity of cigarette butts, and their chemical components, to marine and freshwater
fish. Tob. Control. 20, i25–i29.
Suárez-Rodríguez, M., Garcia, C.M., 2017. An experimental demonstration that house
finches add cigarette butts in response to ectoparasites. J. Avian Biol. 48, 1316–1321.
Suárez-Rodríguez, M., Macías Garcia, C., 2014. There is no such a thing as a free cigarette;
lining nests with discarded butts brings short-term benefits, but causes toxic damage.
J. Evol. Biol. 27, 2719–2726.
Suárez-Rodríguez, M., López-Rull, I., Macías Garcia, C., 2013. Incorporation of cigarette
butts into nests reduces nest ectoparasite load in urban birds: new ingredients for
an old recipe? Biol. Lett. 9.
US-EPA, 2012. Ecological effects test guidelines. OCSPP 850.3100: Earthworm Subchronic
Toxicity Test [EPA 712-C-016].
Yasoshima, M., Takano, B., 2001. Bradybaena similaris (Férrusac) shell as a biomonitor of
copper, cadmium, and zinc. Bull. Environ. Contam. Toxicol. 66, 239–248.