The Journal of Toxicological Sciences (J. Toxicol. Sci.

)
795
Vol.39, No.5, 795-802, 2014

Original Article

Volatile organic compounds of polyethylene vinyl acetate

plastic are toxic to living organisms
Tingzhu Teresa Meng

North Carolina School of Science and Mathematics, 1219 Broad Street, Durham, North Carolina, USA 27705

(Received May 29, 2014; Accepted August 20, 2014)

ABSTRACT — Volatile organic compounds (VOCs) in polyvinyl chloride (PVC) plastic products readi-
ly evaporate; as a result, hazardous gases enter the ecosystem, and cause cancer in humans and other ani-
mals. Polyethylene vinyl acetate (PEVA) plastic has recently become a popular alternative to PVC since it
is chlorine-free. In order to determine whether PEVA is harmful to humans, this research employed the
freshwater oligochaete Lumbriculus variegatus as a model to compare their oxygen intakes while they
were exposed to the original stock solutions of PEVA, PVC or distilled water at a different length of time
for one day, four days or eight days. During the exposure periods, the oxygen intakes in both PEVA and
PVC groups were much higher than in the distilled water group, indicating that VOCs in both PEVA and
PVC were toxins that stressed L. variegatus. Furthermore, none of the worms fully recovered during the
24-hr recovery period. Additionally, the L. variegatus did not clump together tightly after four or eight
days’ exposure to either of the two types of plastic solutions, which meant that both PEVA and PVC neg-
atively affected the social behaviors of these blackworms. The LD50 tests also supported the observations
above. For the first time, our results have shown that PEVA plastic has adverse effects on living organ-
isms, and therefore it is not a safe alternative to PVC. Further studies should identify specific compounds
causing the adverse effects, and determine whether toxic effect occurs in more complex organisms, espe-
cially humans.

Key words: Volatile organic compound, Polyethylene vinyl acetate, Polyvinyl chloride,
Environmental toxin, Carcinogen, Lumbriculus variegatus

INTRODUCTION of these organic compounds in homes or their ecological

Volatile organic compounds (VOCs) are chemicals
con- taining carbon that have high vapor pressures at
standard condition. VOCs are used as ingredients in a
wide array of household products such as shower
curtains, plastic cups, lunch boxes, etc. Their high vapor
pressures result from low boiling points, which allow
gases to be easily emitted while the products are in
storage or in use. Thus, concentrations of many VOCs are
up to ten times higher indoors than outdoors (Hester et
al., 1995).
VOCs can cause eye, nose, and throat irritation, head-
aches, loss of coordination, nausea, developmental dam-
age as well as damage to the liver and central nervous,
respiratory, and reproductive systems (Lester et al.,
2008). Some VOCs like benzene and formaldehyde are
known to cause cancer in animals and are suspected to
cause cancer in humans (Huff, 2007; Hauptmann et al.,
2009). Howev- er, at present, not much is known about
the health effects
Correspondence: Tingzhu Teresa Meng (E-mail: meng15t@ncssm.edu)
effects (Hester et al., 1995).
Polymers derived from vinyl chloride monomer are
called polyvinyl chloride (PVC) or “vinyl”. Polyvinyl
chloride is used in a plethora of household products from
flooring to wall covering.
The addition of a chlorine molecule differentiates PVC
from other vinyls. However, during combustion, chlo-
rine produces a carcinogen called dioxin that can cause
reproductive and developmental problems, damage the
immune system, and interfere with hormones. Low expo-
sure to dioxin is a health concern because the chemical
first accumulates in the environment, then in plants, in
animals, and finally in humans through food
consumption. In fact, food accounts for 95 percent of
human exposure to dioxin (Viola, 1969; Struciński et al.,
2011). Therefore, the World Health Organization’s
International Agency for Research on Cancer (IARC)
established vinyl chloride as a known human carcinogen
(Aitio, 1999).

Vol. 39 No. 5
796
T.T. Meng
In addition to vinyl chloride, PVC has other harmful
additives including phenol, cyclohexane, decane, methyl
isobutyl ketone, xylene, undecane, ethanol, dipropylene
glycol methyl ether, phthalates, organotins, lead, mercury,
as well as the carcinogenic compounds toluene and ethyl-
benzene (Lester et al., 2008). Therefore, a previous study
suggested an association between PVC flooring and the
development of bronchial obstruction in children
(Jaakkola et al., 1999). Also, Di (2-ethyl hexyl) phthalate
(DEHP) caused irregularities in male sexual
differentiation when given to pregnant rats (Wolf et al.,
1999).
Therefore, stores like Target and Wal-Mart have pres-
ently replaced PVC shower curtains with chlorine- free
vinyls, most commonly polyethylene vinyl acetate
(PEVA) plastic (Lester et al., 2008). PEVA is also phtha-
late-free and biodegradable, which makes it currently a
popular alternative to PVC. The absence of chlorine has
been shown to considerably reduce harmful off-gassing
(Nou et al., 2011).
Polyethylene vinyl acetate (PEVA) is a copolymer of
polyethylene (PE) and ethylene vinyl acetate (EVA), and
is similar to PVC because it is also made from
petrochem- icals (petroleum and natural gas). A study
conducted by SAFC Biosciences in 2006 detected VOCs,
such as aro- matic compounds, alcohols, aldehydes,
alkanes, alkenes, ketones, phenolics, ethers, amines,
sulfides, etc., in PE and EVA bags. Although PEVA is
chlorine-free, it is full of untested chemicals that may not
make it a safe alterna- tive to PVC. Therefore, it is
necessary to test the health and environmental effects of
those VOCs in PEVA plas- tic.
A lot of methods are used to measure concentrations
of dissolved organic carbon (DOC) of a water sam-
ple, among which measuring UV-visible absorbance and
then calculating concentration is a convenient and effec-
tive way because aromatic molecules of DOC produce
vibronic excitations under certain wavelengths of UV
(Khan et al., 2014). For example, naphthalene or anthra-
thene has a UV absorbance at 270 nm. Additionally, the
absorption of radiation corresponds to functional groups
attached to an aromatic ring, and this can include -OH
group at 270 nm, -OCH3 at 269 nm, -CN at 271 nm,
-CO2 at 268 nm, and -COOH at 273 nm (Williams and
Flemming, 1980). Therefore, UV absorbance measure-
ment at 254 nm is usually used as a substitute to esti-
mate organic carbon content in the water (Najm et al.,
1994; Eaton, 1995). However, the value of absorbance at
254 nm particularly depends on the concentrations of
humic acids in water. When the concentration of hum-
ic acids is low, the UV254 may be too low and gener-
ate a comparatively high random error (Wang and Hsieh,
2001). Therefore, Khan et al. (2014) developed a corre-
lation with very high coefficient (R 2 = 0.98) between UV
272 nm absorbance and concentration of DOC of a sam-
ple that had been passed through a filter with 0.45 μm
pores (Knap et al., 1996). The DOC includes purgeable
(volatile) organic compound (VOC) and non-purgeable
organic carbon (NPOC).
Lumbriculus variegatus, better known as blackworms
or California blackworms, are freshwater oligochaetes
that live in the sediment and silt of ponds and lakes. A
normal behavior for L. variegatus is clumping — if sev-
eral blackworms are together, they will intertwine their
bodies into a ball. In nature, numerous lumbriculus con-
gregate and form large clumped colonies of up to 2,000
individual worms. This allows for increased metabo-
lism, stabilization of body temperatures, effective gas
exchange, and collaborative food sharing (Leppanan and
Kukkonen, 1998). Moreover, clumped worms burrow
their anterior ends in the sediment and extend their poste-
rior up into the overlaying water. This orientation not only
allows for effective gas exchange via dorsal blood
vessels, but also allows for rapid response upon stimuli,
to avoid dangers such as predation and acute toxicity
(Drewes and Fourtner, 1989; Drewes, 1999; Ding et al.,
2001).
L. variegatus is an important component of fresh-
water food webs in Europe and North America because
they feed on microorganisms and organic material, and
is a major food source for fish. Volatile organic chemi-
cals from deposited plastics end up in sediment of lakes
and rivers through wind and rain run-off. These toxins
diffuse through Lumbriculus’ skin and bio-accumulates
in the blackworm, then in its predators and eventually in
humans. As a result the health of L. variegatus is a vital
indicator of the current and future health of the ecosys-
tem.
This research was conducted to compare effects of
VOCs in both PEVA and PVC on the oxygen intake,
activity and social behavior of the oligochaete Lumbric-
ulus variegatus to determine whether PEVA is toxic and
whether it is a healthy alternative to PVC, and to provide
further information about the health and ecological risk of
these plastics.
MATERIALS AND METHODS
Preparation of PEVA and PVC solutions
To prepare the solution containing volatile organic
com- pounds from PEVA plastics, twenty squares
measuring 2 inches by 2 inches were cut from a 100%
PEVA plastic shower liner. 200 mL of distilled water was
 797

VOCs of PEVA plastic are toxic

poured into a water bath, and then the pieces of
plastic were even-
 ly spread out in the water. The cover of the water bath
was sealed with tape to prevent gases from escaping. The
water bath was heated to 66 degrees Celsius, and kept at
this temperature for two hours, and then was turned off
and left overnight to allow the VOCs condense. The same
procedure was repeated to obtain the solution containing
VOCs from a 100% PVC plastic shower liner.
Measuring concentrations of DOC in six
solutions
Six solutions, including one distilled water, one tap
water, the PEVA solution, the PVC solution, one lake
water (from North Carolina, USA), and one pond water
(from North Carolina, USA), were separately filtered
with sterile and endotoxin-free polyethersulfone filters
with 0.45 μm pores (Whatman TM ), and then read UV
absorbance at 272 nm with NanoDrop ND-1000 V3.3.0
(Thermo Scientific). The settings were as follows: UV-
Vis, 1.0 Max absorbance, Normalize ON, and HiAbs ON.
Next, the concentrations were calculated with the follow-
ing equation:
DOC (mg/L) = 518.93 × Absorbance (272 nm) + 1.065
(Khan et al., 2014)
Obtaining and maintaining L. variegatus
L. variegatus from the same source was purchased
from Carolina Biological Supply Company. Each of
these worms was at a young age — just two weeks old—
and measured 1 to 2 inches long. It was not necessary to
account for gender in the experiment because each worm
has both male and female sex organs, and reproduces
through asexual fragmentation. Also, it was not necessary
to feed the worms because the blackworms are advan-
tageous at long-term exposure without feeding (Sardo
and Soares, 2010) and this experiment did not require
the worms to be maintained for an extended period. The
experimental condition for these worms was at room tem-
perature (25°C) in PEVA, PVC or distilled water.
Determining what concentrations of solutions
are used
A LD50 test was performed prior to the experiment to
determine the toxicity of both PEVA and PVC plastics as
well as the appropriate concentrations to use in the actual
experiment. When establishing a LD50 test, a serial dilu-
tion is used to drop the concentration of the substance to
levels of interest. In this experiment, the concentrations
100%, 10%, 1%, 0.01% and 0.001% of the stock solution
were desired. Thus, the five desired concentrations were
diluted and poured into five dishes, and then ten black-
worms were put in each dish.
After 24-hr, a blackworm died in the 100% PEVA
solution and another blackworm died in the 100% PVC
solution. The blackworms in the 100% solutions did not
clump and tended to be less active than the ones in the
diluted solutions. Although the mortality rates of Lum-
briculus during the LD50 were very low, the VOCs in the
100% solutions did prove to have adverse effect, so the
100% stock solutions from both PEVA and PVC were
chosen to be used in the experiment.
Measuring oxygen intakes
Since the goal of this experiment was to test conse-
quence when Lumbriculus variegatus was exposed to vol-
atile organic compounds in both PVC and PEVA, three
exposures in different lengths were tested: one day, four
days, and eight days. Each length of exposure had three
petri dishes: PEVA (that is, in PEVA solution), PVC (in
PVC solution), and the control (in distilled water). Using
a graduated cylinder, each petri dish of one exposure was
filled with 15 mL of solution of PEVA, PVC or distilled
water, and then 10 blackworms were transferred into it.
The blackworms in each petri dish were transferred to
an appropriate container using a funnel 24-hr before the
due time of an exposure. Then the opening of the contain-
er was sealed with an oxygen probe. The amount of oxy-
gen in the container was measured for 24-hr with a Ver-
nier oxygen gas sensor. After measuring the oxygen, the
blackworms in each container were transferred back into
their original petri dish, and then the solution was sucked
out with a pipette and replaced with 15 mL of distilled
water, and then transferred back into the container again.
The oxygen during blackworms’ recovery was monitored
for 24-hr to see whether the effects of the exposure to
PEVA or PVC were reversible.
The oxygen intakes were calculated after the amounts
of remaining oxygen (in ppm) in the container were
meas- ured with the Vernier oxygen gas sensor. That is, an
oxy- gen intake at a specific time was determined to be
equal to 210,000 ppm (the oxygen concentration in the
empty container) minus the amount of remaining oxygen
at that time. The ppm was then converted to mg by
multiplying the container’s total volume (317 mL).
Observations of activity, mortality, and social
behavior
Observations of activity, mortality and clumping were
recorded after each exposure and each recovery peri-
od. Activity was measured as follows: Blackworms in
a control group were gently probed with a pipette and
their reaction was recorded as normal. Activity rating
ranged from 1 to 3, where 1 = inactive (the worm did
not react with one touch), 2 = normal activity level (the
worm reacted like the control group with one touch), and
3 = very active (one touch triggered others to react, too).
Interpreting data
After the oxygen intakes were calculated, the trends of
the oxygen intakes in each case and their lines of best fit
were graphed with Microsoft Excel. The slope of the best
fit line was the oxygen change rate per hour (Δmg/hr),
which reflected the health effects when the worms were
exposed to VOCs. The similar analysis was done on the
data from each of the recovery periods.
RESULTS
The UV absorbance of the six solutions (one distilled
water, one tap water, the PEVA solution, the PVC solu-
tion, one lake water, and one pond water) at 272 nm were
as follows: 0.005, 0.009, 0.012, 0.021, 0.023, and 0.015,
and their estimated concentrations (mg/L) were as fol-
lows: 3.66, 5.74, 7.29, 11.96, 13, and 8.85.
For the one day exposure, the oxygen intakes of both
PEVA (average 9,250 mg) and PVC (23,529 mg) groups
were much higher than the distilled water group (3,041
mg) (see Fig. 1). Furthermore, the high oxygen intakes of
both PEVA (average 11,729 mg) and PVC (17,891 mg)
groups did not fully recover after a 24-hr healing period
in distilled water (3,205 mg) (see Fig. 2). The same was
observed from oxygen intake data in four day exposure,
eight day exposure and their 24-hr recovery periods (see
Table 1).
Generally, the PEVA group had more fluctuations than
the control and PVC groups (see Figs. 2 and 3). The
One Day
One Day Exposure
Exposure
32000
28000
y = 27.91x + 23194
24000
(mg)
es 20000
Control
intak 16000
n PEVA
Oxyge 12000 y = 21.38x + 8993.5
PVC
8000
4000 y = 29.06x + 2692.2
0 8000
0 2 4 6 8 10 12 14 16 18 20 22 24
Time (hr)
Time (hr)
control and PVC groups had relatively constant oxygen
intakes.
The change in oxygen intake (Δmg/hr) per hour is
the slope of the best fit line. During the exposures (see
Figs. 1, 3, 5), the slope of the control (distilled water) first
decreased from one day exposure 29.06 to four day expo-
sure -19.74 then remained constant from four day to eight
day exposure. The Δmg/hr of PVC first decreased from
27.91 to 2.87 then increased from 2.87 to 47.59. Mean-
while, the Δmg/hr of PEVA increased from 21.38 to 96.94
then decreased from 96.94 to 9.99 (see Table 2).
When comparing the Δmg/hr of exposure to recov-
ery, PEVA’s rate of oxygen intake first increased in one
day exposure and subsequent recovery (from 21.38 to
106.90), then decreased in the four day set (96.94 to 1.36)
and the eight day set (9.99 to 2.69). PVC’s Δmg/hr (27.91
to 19.47, 2.87 to -38.26, 47.59 to 25.23) decreased from
exposure to recovery. The control (29.06 to -12.72, -19.74
to 14.05, -19.92 to 4.57) decreased then increased (see
Table 2).
Additionally, the blackworms did not clump togeth-
er tightly after four or eight day exposure to either of the
two plastic solutions (Table 3). The number of worms
clumped together mainly decreased with greater time
of exposure. Recovery periods generally had greater
worms clumping together than the exposure period, but
the worms in the PEVA and PVC solutions did not fully
recovery (see Table 3).
The control groups during all periods of exposure and
recovery had normal activity. The activity level of PEVA
and PVC was initially higher than normal and decreased
to below normal by the eight day of exposure (see Table
4).
During all the exposure and recovery periods, all of
One Day Recovery
32000
28000
24000
s(mg)
intake 20000 y = 19.47x + 17657
Contol
16000 y = 106.9x + 10445
Oxygen 12000
PEVA PVC
4000 y = -12.72x + 3357.8
0
0 2 4 6 8 10 12 14 16 18 20 22 24
Time (hr)
Fig. 1. One day exposure. The equation for the trend line is Fig. 2. One day recovery. The dash line crossing the PEVA
above each curve. The trend line does not show up if it
curve is its trend line.
completely overlaps with its corresponding curve. The
same rules apply thereafter.
Table 1. Average Oxygen Intake (mg) Table 2. Change in Oxygen Intake (Δmg/hr)
Exposure Avg. Oxygen Intake (mg)
Exposure (Δmg/hr)
Length of Exposure Control PEVA PVC Length of Exposure Control PEVA PVC
One Day 3,041 9,250 23,529
One Day 29.06 21.38 27.91
Four Days 2,719 10,322 23,271
Four Days -19.74 96.94 2.87
Eight Days 2,420 8,056 16,823
Eight Days -19.92 9.99 47.59

Recovery Avg. Oxygen Intake (mg)

Recovery (Δmg/hr)
Length of Exposure Control PEVA PVC
Length of Exposure Control PEVA PVC
One Day 3,205 11,729 17,891
One Day -12.72 106.9 19.47
Four Days 3,404 8,192 22,893
Four Days 14.05 1.36 -38.26
Eight Days 2,448 8,213 17,513
Eight Days 4.57 2.69 25.23

Table 3. The Amount of Clumping Worms Table 4. Activity Level

Exposure (# Clumping) Exposure (Activity Level)
Length of Exposure Control PEVA PVC Length of Exposure Control PEVA PVC
One Day 10/10 8/10 9/10 One Day 2 2.5 3
Four Days 9/10 5/10 6/10 Four Days 2 1 2.5
Eight Days 8/10 2/10 5/10 Eight Days 2 0.5 1

Recovery (# Clumping) Recovery (Activity Level)

Length of Exposure Control PEVA PVC Length of Exposure Control PEVA PVC
One Day 10/10 9/10 10/10 One Day 2 1.5 1.5
Four Days 9/10 7/10 6/10 Four Days 2 1 1.5
Eight Days 9/10 6/10 4/10 Eight Days 2 0.5 1
Note: 1 = inactive, 2 = normal activity, and 3 = very active
ment of exercise respiratory gas exchange is a valuable
and standard testing of cardiopulmonary exercise stress in
the blackworms were respiratory (that is, their tails curled
patients (Crespo et al., 2009).
and were out of the water) and none of the blackworms
As Giesy (1988) stated, the methods to measure the
died under observation.
effects of stressors in aquatic organisms can be placed
into six categories: 1) Hormones; 2) Energetics; 3)
DISCUSSION
Enzyme
activities; 4) Osmoregulatory electrolytes; 5) RNA/DNA
Oxygen consumption is an effective parameter and protein content; and 6) other substrates. Obvious-
of physical stress ly, measuring oxygen consumption belongs to the sec-
Measure of oxygen consumption is an indirect but ond category, and measuring heat shock protein or imme-
common technique to measure energy expenditure diate-early gene products falls into the fifth category. It
(Levine, 2005) because most of energy in the body of an is no doubt that all of them, including oxygen consump-
organism is produced aerobically, and monitoring energy tion, induction of heat shock proteins, induction of imme-
expenditure is a standard method to measure the effects of diate-early gene products, hormones, and osmoregulatory
stressors in aquatic organisms (Giesy, 1988). Therefore, it eletrolytes, are effective parameters to measure stress.
is reasonable to measure oxygen intakes of the worms to
indicate the stress that they endure. Actually, the rate of
respiration/intake of oxygen of earth-dwelling worms has
been shown to increase by introducing stress such as light
stimuli (Billstein et al., 2002). Moreover, direct measure-
 Four Day Exposure Four Day Recovery
Four Recovery

32000
32000
28000
28000
(mg) 24000 y = 2.87x + 23237 24000 y = -38.27x + 23253
s 20000 (mg)

16000
intake y = 96.94x + 9159.1
Control intakes 20000
Control
16000
Oxygen 12000 en PEVA
Oxyg 12000 y = 1.36x + 8179.7
PVC
8000
PEVA PVC 4000
y = 14.05x + 3271.9
8000 0
0 2 4 6 8 10 12 14 16 18 20
4000 y = -19.74x + 2955.5
Time (hr)
Time (hr)
0

Time (hr)

Fig. 3. Four day exposure. The dash line crossing the PEVA
Fig. 4. Four day recovery. The dash line overlapping the PEVA
curve is its trend line, and the thin line overlapping the
curve is its trend line, and the thin line overlap- ping
control curve is its trend line.
the control curve is its trend line.

Eight Day Exposure Eight Day Recovery

32000
32000

28000 28000

24000
24000 (mg)
es
(mg)
20000 es 20000 y = 25.23x + 17210
k y = 47.59x + 16252 k
Control Control
inta 16000 inta 16000
n n PEVA
PEVA
Oxyge 12000 y = 9.99x + 7935.7 Oxyge
12000
y = 2.69x + 8180.6 PVC
PVC 8000
8000
4000 y = -19.92x + 2659.1 4000 y = 4.57x + 2393.2

0 0
0 2 4 6 8 10 12 14 16 18 20 22 24 0 2 4 6 8 10 12 14 16 18 20 22 24
Time (hr)
Time (hr) Time (hr)
Time (hr)

Fig. 5. Eight day exposure. The dash line overlapping the PEVA Fig. 6. Eight day recovery. The dash line overlapping the
curve is its trend line, and the thin line overlap- ping PEVA curve is its trend line.
the control curve is its trend line.

Both solutions stressed the worms and
irreversibly increased their oxygen intakes
Immediate responses of the blackworms to VOCs were
observed at the LD50 test. That is, a blackworm died in
both the original PEVA stock solution and the original
PVC stock solution. Furthermore, the blackworms in the
stock solutions did not clump and tended to be less active
than the ones in the diluted solutions. Therefore the LD50
test demonstrated that the greater the concentrations, the
more toxic were the effects of the VOCs, as is consist- ent
with toxicity studies while treated blackworms with
caffeine, nicotine, etc. (Bohrer, 2006), and with extracts
from various hosta varieties (Rivera and Myers, 2012).
And, one day, four day and eight day exposures were
compared to see the effect of VOCs on the oxygen intake
of L. variegatus. PVC had the highest oxygen intake in all
of the graphs (total average 15,605 mg), however, PEVA
(9,209 mg) still had very high intake compared to the
control (2,726 mg) (see Figs. 1-6). Therefore, these
results clearly indicated that PEVA, like PVC, contained
toxins that stressed L. variegatus and thus increased their
oxygen intakes.
The 24-hr recovery period after the exposure represent-
ed detoxification. When comparing exposure to recov-
ery, there was not much difference in the average oxygen
intake. The recovery data showed that complete recov-
ery did not occur within the given time period (as shown
from the avg. oxygen intake and clumping). These results
 led to the conclusion that the adverse effects of exposure
are irreversible in a 24-hr period.
In the exposures (see Figs. 1, 3, 5), the slopes of the
control (distilled water) first decreased then remained
constant, which indicated that the blackworms adjust-
ed to the new environment and then continued to take
in the same amount of oxygen. The Δmg/hr of PVC first
decreased then increased. Similarly, Billstein et al. (2002)
showed that when earth-dwelling worms were exposed
to stress such as light stimulus, their oxygen intake lev-
el increased. Thus, the longer blackworms were exposed
to PVC, the more stressed they became. However, Δmg/
hr of PEVA first increased then decreased significantly
(96.94 mg to 9.99 mg), perhaps because the harmful com-
pounds of PEVA, which affected the ability of L. varieg-
atus to aspirate oxygen, were different from the toxins of
PVC.
Both solutions adversely affected the worms’
behavior
As exposure length increased, the number of worms
clumping together decreased. Activity level of worms in
both PEVA and PVC solutions was initially higher than
normal and then decreased to below normal by the eight
day of exposure, which meant that the stress excited the
blackworms’ body systems, and then weakened their abil-
ity to respond. Therefore, there was no doubt that increas-
es in concentration and exposure length to either PEVA or
PVC solution negatively affected blackworms’ social
behavior. A similar phenomenon was observed by other
researchers: Bohrer (2006) noticed that the blackworms
were not or less clumping while living in water treated
with caffeine or nicotine.
Experimental conditions were well controlled
Not all of the blackworms at each exposure length
demonstrated the same behavior (see Table 3, Table 4).
This can be explained by variability or differences among
the worms. Both extrinsic and intrinsic factors may affect
results of experiments. However, extrinsic factors, such
as temperature and pressure, were controlled via
conducting the experiment in standard room conditions.
Intrinsic fac- tors are within individual organisms, such as
age, metabo- lism, and genetic difference. In the
experiment, L. varieg- atus picked for each group were
selected to be all roughly the same length and coloring,
thus age and body size was controlled. Still, genetic
difference between blackworms could have affected
results of exposure to VOCs. How- ever, the genetic
difference among the worms should be small since all of
them were fresh young, were from the same source at the
same time, and were almost the same
size.
Toxic organic compounds are potential health
risk to human and other animals
UV absorption of organic solutes is directly
proportion- al to their concentration in aromatic
compounds (Traina et al., 1990; Chin et al., 1994), and
thus measuring UV- Vis absorbance is a convenient and
effective way to esti- mate concentrations of DOC in
water (Najm et al., 1994; Eaton, 1995; Khan et al., 2014).
The DOC of PVC solution is lower than that of the
lake water (11.96 mg/L vs. 13 mg/L), and higher than that
of the pond water (11.96 vs. 8.85). The DOC of PEVA
solu- tion is close to that of the pond water (7.29 vs.
8.85), and higher than the tap water (7.29 vs. 5.74). These
data indi- cate that toxic organic compounds are possibly
accumu- lated in the environment such as lakes, ponds
etc., repre- senting a potential health risk to human and
other animals. Further steps will include determining
what components in the two plastics are toxic to the
blackworms, and how much they have accumulated in the
ecosystem.
As a side note, the concept Dissolved Organic Matter
(DOM) in the reference Khan et al. (2014) is the same
as the concept Dissolved Organic Carbon (DOC) in this
article because of two reasons: Firstly, samples in the two
studies were passed through filters with 0.45 μm pores
and then were measured UV-Vis absorbance. Secondly,
the practical definition of DOC is the "dissolved" frac-
tion of organic carbon below 0.45 μm (Knap et al., 1996),
which includes VOC and NPOC.
In summary, this research clearly showed that PEVA
has toxic effects on the oxygen intake, social behavior
and activity of Lumbriculus variegatus, even though it is
chlo- rine-free. And, this research proves that there is a
need for further study on the toxicity of VOCs in PEVA
plas- tic, especially to see whether it causes cancer in
humans. In the near future, further studies should
determine what compounds caused these adverse effects,
how much they have accumulated in the ecosystem, and
whether the tox- ic effect occurs in more complex
organisms, especially humans.
ACKNOWLEDGMENTS
This research was supported by North Carolina
Summer Ventures in Science and Mathematics, and River
Center at Duke University. Many thanks to Josh Cannon
for instruction and discussion, and to Dr. Christine Muth,
Brooke Hassett and Chelsea Clifford for their help.
Conflict of interest---- The authors declare that there is iment by using UV–Visible spectrophotometer. International
no conflict of interest. Journal of Environmental Pollution and Remediation, 2, 24-29.
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