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Estimation of arsenic and mercury in fishes from river Ganga for riverine ecosystem health biomonitoring and assessment, J. Inland Fish. Soc. India, 49 (1) : 48-56, 2017

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Concentration of pollutants in fish tissues is a true reflection of their concentration in the ecosystem. In this context, toxic environmental contaminants arsenic and mercury in fishes from river Ganga were investigated by inductively-coupled plasma mass spectrometry (ICP-MS) to assess the level of these contaminants in different stretches of the river. Total arsenic concentrations in Tenualosa ilisha, Sperata seenghala, Amblypharyngodon mola and Puntius sophore were 0.02-2.9 mg kg-1 which is within the permissible limit for human consumption. Mean mercury content was below the permissible level (0.003-0.05 mg kg-1) in all fishes analyzed from different stretches. This biomonitoring study showed that river Ganga, in the indicated and adjoining stretches, appears to be free from arsenic and mercury contamination. This study demonstrated the dual benefits associated with using fish as biomonitors; it serves as a tool for riverine ecosystem health monitoring and also provides information on food safety.
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ESTIMATION OF ARSENIC AND MERCURY IN FISHES FROM RIVER
GANGA FOR RIVERINE ECOSYSTEM HEALTH BIOMONITORING AND
ASSESSMENT
BIMAL PRASANNA MOHANTY, SATABDI GANGULY, ARABINDA MAHANTY,
TANDRIMA MITRA, PRASENJIT PARIA, BIJAY KUMAR BEHERA, BASANTA KUMAR DAS
ICAR- Central Inland Fisheries Research Institute, Fishery Resource and Environmental Management
Division, Biochemistry Laboratory, Barrackpore, Kolkata 700120, India
Email of corresponding author: bimal.mohanty@icar.gov.in
(Received : 22.09.2017; Accepted : 12.10.2017)
Concentration of pollutants in fish tissues is a true reflection of their concentration in the ecosystem. In this context,
toxic environmental contaminants arsenic and mercury in fishes from river Ganga were investigated by
inductively-coupled plasma mass spectrometry (ICP-MS) to assess the level of these contaminants in different stretches
of the river. Total arsenic concentrations in Tenualosa ilisha, Sperata seenghala, Amblypharyngodon mola and
Puntius sophore were 0.02-2.9 mg kg-1 which is within the permissible limit for human consumption. Mean mercury
content was below the permissible level (0.003-0.05 mg kg-1) in all fishes analyzed from different stretches. This
biomonitoring study showed that river Ganga, in the indicated and adjoining stretches, appears to be free from arsenic
and mercury contamination. This study demonstrated the dual benefits associated with using fish as biomonitors; it
serves as a tool for riverine ecosystem health monitoring and also provides information on food safety.
Key words: Arsenic, ICP-MS, Fish biomonitors, Food safety, Mercury, Riverine ecosystem health biomonitoring
J. Inland Fish. Soc. India, 49 (1) : 48-56, 2017
46
Introduction
Periodic monitoring of the water quality, especially
for toxic elements like arsenic and mercury in the
rivers is necessary to evaluate the riverine ecology
and its suitability and capacity for sustaining life in
the entire food chain, from aquatic flora and fauna
to humans. The environment, in an aquatic
ecosystem, embodies the total physical and
chemical factors that exert an effect upon the biotic
communities; however, despite its essential
biological character, environmental quality is often
assessed in terms of physical and chemical
parameters only (Bere and Nyampuingidza, 2013).
The biotic components can serve as important tools
for assessment of health of an aquatic ecosystem
as water pollution leads to bioaccumulation of the
pollutants in fish and other aquatic organisms.
Therefore, fish and shellfish based approaches are
more appropriate for assessing contamination of
the aquatic ecosystem (Trautwein et al., 2013).
Using fish for aquatic pollution monitoring has dual
advantages; firstly, it can be used for assessing the
level of contamination of toxic elements affecting
the flora and fauna, secondly assessment of
accumulation of these elements in fish tissues
provides important information from food safety
point of view.
Contamination of the aquatic ecosystems like lakes,
rivers and underground water with toxic metals/
metalloids like mercury (Hg) and arsenic (As) is
an issue of environmental concern and has received
worldwide attention (Sinha et al., 2007;
Bhattacharyya et al., 2010, Chowdhury et al.,
2015; Mohanty et al., 2015). Industrial and
domestic waste water, agricultural pesticides,
xenobiotics etc. are chief sources of environmental
contamination which contribute to the river pollution
(Shah et al., 2009; Samanta 2013).
Aquatic pollution leads to bioaccumulation of these
pollutants in fish. Therefore, fish is used as an
indicator of heavy metals and other pollutants in
the aquatic ecosystem (Djedjibegovic et al.,
2012). Groundwater contamination with arsenic
has become a major health problem in many
countries of the world; worst affected ones being
West Bengal in India and the Bangladesh. Chronic
exposure through prolonged drinking of
arsenic-rich water leads to arsenicosis and results
in various pathophysiology that include alteration
of skin colour, hard patches on palms and soles of
feet, cancers of the skin, bladder, kidney and lung,
and vascular diseases in humans (Guha-
Mazumder, 2008). Although human exposure is
mainly through arsenic-contaminated water, recent
studies have shown that besides drinking waters,
food chain is a major contributor to arsenicosis
(Rahman et al., 2008).
Mercury is one of the most hazardous heavy metal
toxicants. Mercury in nature is readily convertible
to methyl mercury which is the most toxic form of
mercury and fish and shellfishes are highly affected
by methyl mercury contamination (Mahaffey,
2005). Studies have revealed that exposure to toxic
levels of mercury leads to neurological
abnormalities, cognitive impairment and
behavioural disturbance (Nascimento et al.,
2008). Low dose mercury toxicity has also been
found to affect various organ systems like renal
system, reproductive system, cardiovascular and
the immune system (Zahir et al., 2005).
Biomonitoring, the use of biological responses to
assess changes in the environment, is a valuable
assessment tool necessary for water quality
monitoring. It provides the direct evidences of
alterations that have occurred in the ecosystem due
to environmental pollution (Zhou et al., 2008).
Biomonitoring involves the use of bioindicator
species; generally, the benthic macro invertebrates,
fish and algae etc. are used as indicators. The
presence or absence of the indicator species is
indicative of the environmental conditions as they
integrate the effects of different stressors and
provide information on the overall impact (Winner
and Bewley, 1978; O'Conner et al., 2015).
Periphyton assemblage, macroinvertebrate
assemblages are useful as indicators.
Macroinvertebrate assemblages like gastropod
and bivalve molluscs are good indicators of
localized conditions as these sentinel organisms
have limited migration (Barbour et al., 1999).
Fishes, on the other hand are good indicators of
long-term effects and broad habitat conditions, as
they are relatively long-lived and mobile. Fish
assemblages generally include a range of species
that represent a variety of trophic levels
(omnivores, herbivores, insectivores, planktivores,
piscivores). They tend to integrate effects of lower
trophic levels; thus fish assemblage structure is
reflective of integrated environmental health (Karr
et al., 1986). Moreover, fish are at the top of the
aquatic food web and are consumed by human,
making them more important for assessing
contamination. Keeping this in view, the present
biomonitoring study has been carried out using fish
bioindicator for rapid assessment of the level of
arsenic and mercury contamination in the river
Ganga, the longest river system in India.
Materials and methods
Sample collection
Amblypharyngodon mola, Puntius sophore and
Sperata seenghala were collected from Kanpur,
47
AS AND HG MONITORING IN FISHES FROM RIVER GANGA
Allahabad (UP) and Seoraphuli (WB) where as
Tenualosa ilisha was collected from Seoraphuli
and Diamond Harbour (WB) (Fig. 1). Edible
muscle tissues were collected from different size
groups (small, 200-400 g; medium, 800-1000 g;
large, 1400-1600 g) of Tenualosa ilisha and
Sperata seenghala. Eight individual fish samples
were analyzed in triplicate. For the small indigenous
fishes (SIFs) a single pooled sample was prepared,
containing upto hundred individual fish. The (SIFs)
Amblypharyngodon mola and Puntius sophore,
degutted whole fishes were taken for analysis. The
minced fishes were kept overnight in a hot air oven
at 120 °C and then the dried samples were
powdered in a mixer grinder and stored in aluminum
foils for further use.
Mineral analysis
Arsenic and mercury were analyzed by Inductively
Coupled Plasma Mass Spectrometry (ICP-MS)
following standard protocol as described earlier
(Mohanty et al., 2012a). Briefly, a microwave
assisted digestion was carried out to achieve a
shorter digestion time. Dried fish powders (0.5 g),
in triplicate, were placed in glass digestion bombs
and 3 ml suprapure HNO3 (E. Merck) was added
to the samples. The bombs were firmly closed and
put in the microwave oven (MDS-Anton Paar,
Multiwave 3000) for digestion under controlled
pressure. The basic program of the microwave
digestion is given in Table 1. After digestion, the
glass bombs were cooled and the mineralized
samples were diluted to 50 ml with milli Q water
and stored in refrigerator till ICP-MS (Thermo
Fisher X Series 2) analysis. Arsenic and mercury
were directly analyzed in ICP-MS without
diluting the mineralized samples. A commercially
available multi-element stock standard solution was
used, after appropriate dilution for instrumental
calibration and sample spiking (1.09494.0100, E.
Merck). ICP-MS operation and measurement
conditions are given in Table 2.
Statistical analysis
Mean element concentrations in fish species
collected from different sites of river Ganga were
tested for significance by analysis of variance
(ANOVA) with multiple comparisons Tukey’s test
using SPSS 16.0 software. Significance was
established at p < 0.05.
Results and discussion
Four different species of fishes were selected for
the present study keeping in view their relevance
in aquatic pollution monitoring as well as
commercial importance as food fishes. In our
earlier study, we have reported the micronutrient
composition of these food fishes (Mohanty et al.,
2016) and in the present study arsenic and
mercury concentration was evaluated for food
48
MOHANTY et al.
Fig. 1. Map showing the sampling sites along the course
of river Ganga.
safety issues. Most of the environmental
contaminants accumulate in fat tissues of animals
(Zahir et al., 2005); therefore, it is necessary to
measure the level of contaminants in oil-rich fishes
like Tenualosa ilisha which contains about
8-15% fat or oils of the total body weight
(Mohanty et al., 2012b). This fish also
contributes a major share to the riverine catch and
is in good demand as a food fish; moreover, being
a migratory fish the concentration of arsenic and
mercury in this fish can reflect the status of the
level these contaminants in the zone of its
migration. The toxic environmental contaminant
arsenic has been a major public health problem in
the Indian subcontinent, especially in West Bengal
in India and Bangladesh. Hilsa is produced and
consumed in largest quantity in this region also.
Therefore, it is quite pertinent to monitor the level
of arsenic accumulation in hilsa meat.
Carnivorous fishes serve as good models for
biomonitoring of toxic environmental contaminants
as these fishes accumulate the toxicants directly
from the environment and also through the food
chain. The giant river-catfish Sperata seenghala
is available in most stretches of river Ganga and is
also the major catfish contributing to the riverine
catch; therefore it was included in this study
(Mohanty et al., 2012a). The two SIFs, A. mola
and P. sophore are micronutrient dense and are in
great demand these days for their nutritive values
(Mohanty et al., 2010). Therefore, these small
fishes were also selected for monitoring the level
of arsenic and mercury.
The sampling sites were chosen considering the
vulnerability of the river to receive high amounts
of pollutants from these places. Kanpur is a
major industrial township situated on the bank of
river Ganga. So, the river gets contaminated with
significant amount of industrial effluents in Kanpur.
Allahabad is located at the confluence of two
rivers, Ganga and Yamuna, both running for very
long distances carrying bulk of pollutants with them.
The other site, Seoraphuli is a suburban township
of Kolkata along the river Ganga. Diamond
Harbour is very close to the place where the river
meets the Bay of Bengal. Thus, the selected sites
represent the middle and lower stretches of the
river where the pollution load is likely to be high
due to both industries and anthropogenic
activities (Samanta, 2013).
ICP-MS is an advanced technology that can
detect metals at a concentration of the order of
10-12 (parts per billion, ppb) with simultaneous multi
elemental measurement capability, wide linear
dynamic range and simpler spectral interpretation
(Djedjibegovic et al., 2012). Joint FAO/WHO
(2010) (FAO/WHO, 2010) has certified ICP-MS
as a better technique due to its high sensitivity
Table 1. Microwave digestion programme for pulverized
fish samples
Step Power Ramp Hold time
(W) time (min) (min)
1 300 05.00 05.00
2 400 10.00 10.00
3 600 10.00 05.00
Table 2. ICP- MS operating and measurement
conditions for quantification of As and Hg
Operating Conditions
Spray chamber Cyclonic Quartz
RF Power (W) 1398
Plasma Gas Flow (L min-1) 13.0
Nebuliser gas flow rate (L min-1) 0.98
Auxilary gas flow rate (L min-1) 0.5
No. of replicates 3
Isotopes 75As, 202Hg
Delay Time (sec) 5
Sample Flush Time (sec) 30
Flush pump rate (rpm) 100
Analysis pump rate (rpm) 40
49
AS AND HG MONITORING IN FISHES FROM RIVER GANGA
without derivatization; thus, ICP-MS is the
technique of choice in element analysis.
The mean arsenic content of T. ilisha at different
sampling sites was 2.9 mg kg-1 at Seoraphuli and
1.9 mg kg-1 at Diamond Harbour; the same for S.
seenghala was 0.04 mg kg-1 at Kanpur, 0.19 mg
kg-1 at Allahabad and 0.05 mg kg-1 at Seoraphuli,
(Table 3). The mean arsenic content in A. mola
and P. sophore collected from different sampling
sites were 0.11 and 0.10 mg kg-1, respectively
(Table 4). ANOVA analysis along with multiple
comparison Tukey’s post hoc test showed that
mean arsenic content of T. ilisha from sampling
site Seoraphuli was significantly higher than the
other site Diamond Harbour, WB (p < 0.05).
Sorting in order, from highest to lowest,
concentration of arsenic, the fishes can be ranked
as T. ilisha > S. seenghala > P. sophore >
A. mola.
The arsenic content of these fishes were compared
with the data available for other fishes in the
literature (Falco et al., 2006; Saei-Dehkordi
et al., 2010, Nadal et al., 2008). Arsenic
accumulation in muscle tissue of different species
is related to feeding behavior, specific physiology
of the fish species, dietary habits which further
varies with geographical location (Falco et al.,
2006). The mean arsenic content of T. ilisha at
different sampling sites was found to be
2.4 mg kg-1 wet weight. Saei-Dehkordi et al.
(2012) reported the total arsenic content of
fifteen most consumed fish species from the
Persian Gulf to be in the range of 0.156-0.834 mg
kg-1 which comprises fishes like barracuda,
cobia, tuna, seabream, herring, pomfret etc.
However, the mean arsenic concentration in
T. ilisha was found to be lower than that reported
by Falco et al. (2006) who measured the arsenic
concentration in fourteen edible marine species
from Catalonia, Spain. Among them sardine
(3.53-3.94 mg kg-1), anchovy (3.93-5.42 mg kg-
1), hake (3.22-4.55 mg kg-1), sole (4.55-8.40 mg
kg-1), shrimp (3.85-8.76 mg kg-1) and red mullet
(15.39-17.77 mg kg-1) have higher arsenic
concentration than the present concentration level
in T. ilisha. A similar study from the
Mediterranean Sea reported higher arsenic
content in sardine (4.57 mg kg-1) (Nadal et al.,
2008).
The organic forms of arsenic present in fish when
ingested undergo very little biotransformation and
Table 3. As and Hg level (mg kg-1 wet weight) in Tenualosa ilisha and Sperata seenghala from river Ganga
Fish As (mg kg-1 wet weight) Hg (mg kg-1 wet weight)
Small medium Large Average Small Medium Large Average
T. ilisha
Seoraphuli, WB 2.2±0.09b3.3±0.09b3.2±0.11b2.9±0.60b0.001±0.00b0.015±0.00b0.001±0.00b0.005
±0.00a
Diamond 2.1±0.02b2.3±0.00a1.4±0.02c1.9±0.47a0.001±0.00b0.002±0.00a0.003±0.00b0.002
Harbour, WB ±0.00c
S. seenghala
Kanpur, UP 0.05±0.02a0.02±0.01a0.05±0.00a0.04±0.01a0.07±0.01a0.03±0.00a0.04±0.00a0.04
±0.02a
Allahabad,UP 0.5±0.05b0.05±0.02b0.04±0.01b0.19±0.26b0.02±0.00b0.05±0.00b0.008±0.00b0.02±0.02b
Seoraphuli, WB 0.07±0.04a0.04±0.01c0.04±0.00b0.05±0.0b0.06±0.00c0.009±0.00b0.2±0.00c0.089±0.09c
a, b, c Different letters in superscript indicate that mean values in a row are significantly different according to Tukey’s
multiple range test (p < 0.05).
50
MOHANTY et al.
are excreted entirely unchanged (FAO/WHO,
2010). Among the four fish species analyzed in
the present study, T. ilisha showed highest arsenic
content; however, it is lower than many
commercially important fishes like sardine,
anchovy, hake, red mullet etc. reported in earlier
studies (Falco et al., 2006; Nadal et al., 2008).
Review of literatures indicated that in fish flesh,
collected both from fresh water and marine
systems total arsenic was recorded in the range of
0.00 to 3.50 mg kg-1 on fresh weight basis. The
observed arsenic content was mostly contributed
by different organic forms and the inorganic forms
were recorded as the meagre fractions and mostly
in the level of traces to 0.50% of the total content
(WSDE 2002; Moreau et al., 2007). The Joint
FAO/WHO Expert Committee on Food Additives
(FAO/WHO, 2010) legislated the maximum daily
tolerable intake of organoarsenic to be 0.05 mg
kg-1 of body weight and for a person weighing
about 60 kg, the permissible daily intake would
be 3.0 mg. In the present study, total arsenic
concentration in S. seenghala, A. mola and
P. sophore ranged from 0.02-0.14 mg kg-1 in
comparison to 0.060-4.72 mg kg-1 for freshwater
fishes as reported by FAO/WHO Expert
Committee, 2010. Thus, the arsenic
concentrations in fish from the river Ganga at the
specified stretches are well within the permissible
limit (FAO/WHO, 2010).
Mercury concentration in S. seenghala at
different sampling sites was 0.04 mg kg-1 at Kanpur,
0.02 mg kg-1 at Allahabad, 0.09 mg kg-1
Seoraphuli and in T. ilisha it was 0.005 mg kg-1 at
Seoraphuli and 0.002 mg kg-1 at Diamond Harbour
(Table 3). The mean mercury concentration in
A. mola and P. sophore were 0.003 and
0.04 mg kg-1, respectively (Table 4). The mean
mercury content of S. seenghala and P. sophore
showed significant difference among all sampling
sites (p < 0.05). Mercury bioconcentration in fish
muscle decreased in the order S. seenghala >
P. sophore > A. mola > T. ilisha; however, in all
cases the tissue mercury level was below the
permissible limit. The mean mercury content of
S. seenghala (0.05 mg kg-1) and P. sophore
(0.04 mg kg-1) was found to be higher than the
other two species studied which could be due to
lesser efficacy of certain fish species to remove
toxicants as compared to other fish species
(Agarwal et al., 2007). Moreover, it is known that
the muscle tissue of predatory fish contains
significantly higher level of total mercury than the
muscle tissue of non-predatory fish indicating
bioaccumulation and biomagnifications of the
element through food chain (Vieira et al., 2011).
Thus, the mercury contaminations of the studied
fish samples were contributing much less amount
of the contaminant since the average per-day fish
consumption is about 50-100g while the daily
permissible level, as recommended by FAO/WHO
2010, is 0.05 mg day-1 for a person having
70 kg-1 body weight-1; so are safe for human
consumption. The mercury concentrations of
S. seenghala and P. sophore were notably lower
Table 4. As and Hg level (mg kg-1 wet weight) in small
indigenous fishes (SIFs) A. mola and P. sophore from
the river Ganga
Fish As Hg
(mg kg-1 (mg kg-1
wet weight) wet weight)
A. molad
Kanpur, UP 0.1±0.052a0.004±0.00a
Allahabad, UP 0.09±0.00a0.001±0.00a
Seoraphuli, WB 0.14±0.00a0.01±0.003b
P. sophored
Kanpur, UP 0.06±0.00a0.02±0.00a
Allahabad, UP 0.15±0.00b0.08±0.00b
Seoraphuli, WB 0.11±0.00c0.01±0.00c
a, b, c Different letters in superscript indicate that mean
values in a row are significantly different according to
Tukey's multiple range test (p < 0.05).; danalyzed in
triplicate.
51
AS AND HG MONITORING IN FISHES FROM RIVER GANGA
than the mean mercury concentration of fourteen
edible fish species from Catalonia, Spain
(Falco et al., 2006). In predatory fish like
mackerel, the total mercury content was reported
to be 0.0956 mg kg-1 in chub mackerel and 0.1715
mg kg-1 in horse mackerel (Vieira et al., 2011).
The total mercury concentration of fifteen most
consumed fish species from Persian Gulf was
reported to be 0.120-0.527 mg kg-1
(Saei-Dehkordi et al., 2010) which is higher than
the mercury concentration found in the present
study. Similar observations (mean mercury
concentration 0.050-0.401 mg kg-1) were made
by Djedjibegovic et al. (2012) in six fish species
collected from different sampling sites along the
river Neretva in Bosnia and Herzegovina. Agarwal
et al. (2007) studied the mercury content in
different fish species in river Gomti, which is a
tributary to the river Ganga, India and found that
the mercury content in some of the fishes like
Mastocembelus armatus and catfish
Clarias batrachus to be as high as 0.2 mg kg-1.
Sinha et al. (2007) have reported the mercury
content of sixty seven fish samples collected from
river Ganga at Varanasi, India. The mercury
content of fishes in their study ranged from not
traceable (NT) to 91.679 ppm (annual mean 2.638
± 1.675 ppm); out of which, Puntius sophore
was found to contain 0.064-0.162 ppm mercury
which is very similar to the values reported in the
present study. Bhattacharya et al. (2010) have
reported the total mercury content in fishes
collected from East Calcutta wetlands and Titagarh
sewage-feed aquaculture in West Bengal, India and
total mercury content in different fish species in
their study was 0.073-0.94 mg kg-1 of wet sample
which is higher than the mean mercury content of
the fishes observed in the present study. Thus, the
present study shows that fishes are free from
mercury toxicity and the river is free from mercury
pollution in the studied stretches (middle and
lower).
The concentration of total arsenic and mercury in
the T. ilisha, S. seenghala, A. mola and P.
sophore are within permissible limits (FAO/WHO,
2010), suggesting that the river Ganga is free from
arsenic and mercury contamination in the indicated
stretches. Arsenic speciation studies are necessary
to be carried out to determine the level of organic
and toxic inorganic forms in the fish meat as more
of organic form is not a problem.
Conclusion
This biomonitoring study in fish showed that the
river Ganga, in the indicated and adjoining stretches
i.e. the middle and lower stretches, is free from
arsenic and mercury contamination. However,
periodic monitoring of the river is necessary as they
are dynamic systems and the aquatic environment
is likely to be affected with spatio temporal
changes in anthropogenic activities and natural
processes. Moreover, speciation studies are also
necessary to be taken off for arsenic as the
toxicity varies with different forms.
Acknowledgements
This work was supported by the Indian Council
of Agricultural Research (ICAR) under Fisheries
Science Division Outreach Activity (#3) on
Nutrient profiling and evaluation of fish as a
dietary component. The authors thankfully
acknowledge the help from M/s ThermoFisher
Scientific, Powai, Mumbai for the ICP-MS
facility. The financial assistance to SG, AM, TM,
PP by ICAR is gratefully acknowledged.
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... Previous studies also reported higher arsenic content comparing to present study in fish from Bangladesh (Raknuzzaman et al. 2016;Ahmed et al. 2019;Rahman et al. 2012). Fish from India has been found to contain arsenic as 0.35 ± 0.08 mg kg −1 in L. rohita from tropical wetlands (Kumar and Mukherjee 2011); 2.9 ± 0.6 mg kg −1 in hilsa shad from Ganga river of India (Mohanty et al. 2017). Arsenic concentration has been recorded in higher magnitude comparing to present study in previous studies for fish from Oman (Sadeghi et al. 2019), Arabian Gulf (Kamal et al. 2015), Persian Gulf (Cunningham et al. 2019), and Pakistan (Shah et al. 2009). ...
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