Brinati et al., 2016

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Electronic supplementary material The online version of this 
article (doi:10.1007/s00128-016-1910-8) contains supplementary 
material, which is available to authorized users.
 
 Jerusa Maria Oliveira
oliveira.jerusam@gmail.com
1 Federal University of Viçosa, Vicosa, Brazil
Received: 21 July 2015 / Accepted: 26 August 2016
© Springer Science+Business Media New York 2016
Low, Chronic Exposure to Endosulfan Induces Bioaccumulation 
and Decreased Carcass Total Fatty Acids in Neotropical Fruit 
Bats
Alessandro Brinati1 · Jerusa Maria Oliveira1 · Viviane Silva Oliveira1 · 
Mirlaine Soares Barros1 · Bruno Marques Carvalho1 · Luciane Silva Oliveira1 · 
Maria Eliana Lopes Queiroz1 · Sérgio Luiz Pinto Matta1 · 
Mariella Bontempo Freitas1
Bull Environ Contam Toxicol
DOI 10.1007/s00128-016-1910-8
being hazardous to non-target animals and to the environ-
ment (Agbohessi et al. 2014 ). However, in some regions, 
EDS is still currently allowed, and even in countries where 
the insecticide is being phased out, monitoring data for pes-
ticides might be difficult (Van Dyk and Pletschke 2011). In 
developing countries, this broad spectrum insecticide can 
still be used clandestinely, including in Brazil (Carneiro 
et al. 2015), where it was officially discontinued in 2013. 
Since then, endosulfan concentrations have been quanti-
fied in vegetables in Brazil (Paulino et al. 2014 ; Carneiro 
et al. 2015) and this endosulfan is generally considered to 
be a global pollutant (Kuvarega and Taru 2007 ). EDS mode 
of action in mammals involves the inhibition of the neu-
rotransmitter gamma-aminobutyric acid (GABA), causing 
cell depolarization and overstimulation to the nervous sys-
tem (Coats 1990). This mode of action may interfere with 
the metabolism of carbohydrates (Kalender et al. 2004; 
Thangavel et al. 2010) as well as hormone secretion, caus-
ing hyperglycemia and decreased testosterone, among other 
metabolic disorders (Singh and Pandey 1990; Saiyed et al. 
2003).
Although there is a growing literature regarding the toxic-
ity of organochlorines, little is known about its effects on the 
energy metabolism of wild animals continuously exposed to 
pesticides. In bats, the bioaccumulation of organochlorine 
pollutants has been reported in several species from temper-
ate areas (Clark Jr. 2001; O’shea et al. 2001; Allinson et al. 
2006; Kannan et al. 2010) and may be related to declines 
on bat populations living near fruit crops treated with pesti-
cides (Allinson et al. 2006; Dennis and Gartrell 2015). Most 
toxicological studies, however, do not involve Neotropical 
bat species. In the tropical rainforest ecosystems, habitat 
loss and forest fragmentation as associated with agricul-
tural practices represent a primary threat to wildlife. The 
great fruit eating bat (Artibeus lituratus) can often be seen 
Abstract We investigated the effects of the insecticide 
endosulfan on energy metabolism and its possible accu-
mulation in fruit bats. Adult male bats (Artibeus lituratus) 
were exposed for 35 days, when they were offered fruit 
treated with endosulfan (E) and adhesive spreader (AS) in 
the following concentrations (g/L): 0.0; 0.0 (Control), 0.0; 
0.015 (AS), 1.05; 0.015 (E1), 2.1; 0.015 (E2). Concentra-
tions used were those recommended by the manufacturer 
for fruit crop application (E1) or twice this value (E2). E1 
bats showed decreased plasma glucose concentration. Car-
cass fatty acids were decreased in E1 and E2 bats. Endo-
sulfan bioaccumulation was observed in both liver and adi-
pose tissues from E1 and E2 bats. These results indicate 
that the chronic exposure of fruit bats to environmentally 
relevant concentrations of endosulfan can lead to signifi-
cant bioaccumulation beyond control and also decreased 
fatty acid content, which may impair the health of this 
important seed disperser in neotropical forests.
Keywords Artibeus lituratus · Carcass fatty acids · 
Gas chromatograph · Metabolism · Organochlorine
The organochlorine insecticide and acaricide endosulfan 
(EDS) has been used worldwide to control pests in agricul-
ture until recently, when it was banned in 63 countries for 
1 3
2 Bull Environ Contam Toxicol
were removed, weighed and stored at −20°C. Plasma glu-
cose concentration was determined by the glucose-oxidase 
enzymatic method (GLUCOX 500, DOLES). The hepato-
somatic index (HSI) was calculated using the following for-
mula: HSI = Liver weight (g)/body weight (g) x 100.
Concentrations of liver and muscle glycogen were deter-
mined according to Sjörgren et al. (1938) and quantified in 
spectrophotometer (λ = 620) (Shimadzu). Dextrose was used 
for calibration of the assay and the results are expressed in 
µmol glicosil-unidades/g. Total liver and muscle (breast, hind 
limb and forelimb muscles) protein content were performed 
by the colorimetric assay (BCA Protein Assay Reagent kit, 
PIERCE). Total lipids of the liver, muscles (breast and limbs) 
and adipose tissue, as well as carcass fatty acids, were deter-
mined gravimetrically (Folch et al. 1957).
Chemical analysis of endosulfan concentrations were per-
formed in extracts from the diet and bats liver (1 replicate) 
and adipose tissue (2 replicates) (1 g wet weight) from one 
or pooled animals from all different groups (Table 3). Fruit 
EDS concentration were measured in the fruit peel after 
treatment. Fruit and tissue samples (4 g each) were obtained 
from the method of solid–liquid partition extraction at low 
temperature (ESL-PBT), optimized for extraction of endo-
sulfan. Samples were homogeneized with water and ace-
tonitrile (3:8 v/v) and stored at −20°C for 4 h. Following 
phase separation, the organic liquid phase was filtered with 
a paper previously rinsed with cooled acetonitrile contain-
ing 1 g of sodium sulfate anhydrous. The volume of each 
sample extract obtained was concentrated and adjusted to 
5 mL with acetonitrile. Quantification of the active ingredi-
ent in the extracts of adipose and liver tissues was performed 
by the method of internal standardization. The standard 
curve was prepared with endosulfan at a known concen-
tration diluted with acetonitrile. We prepared two standard 
curves with endosulfan at a known concentration, one for 
tissue samples and the second for quantification in the diet. 
To these solutions and extract samples we added 0.1 mL of 
methyl parathion (10 g L−1), used as internal standard. Chro-
matography analysis was performed using a gas chromato-
graph (GC) (Shimadzu, GC—17A) with an electron capture 
detector (ECD). The chromatographic separation was car-
ried out through a stationary phase capillary column (5 % 
diphenyl, 95 % dimethylsiloxane; 30 m × 0.25 mm of inner 
diameter and 0.1 μm in film thickness). The column tem-
perature was set at 150°C (2 min) with heating from 20°C/
min up to 190°C (3 min) followed by heating 20°C/min to 
the final temperature of 280°C (2 min). The total duration 
of the analysis was 13.5 min. Nitrogen was used to drag the 
gas flow rate of 1.2 mL/min. The injector and the detector 
temperatures were 280 and 300°C. The sample volume was 
1 μL and all division flows were 1:5. Endosulfan concentra-
tions were identified by comparing the compound retention 
time to the pattern for this chemical. Analytical curves were 
foraging in these areas. This species plays a key role in seed 
dispersal, which is crucial for forest regeneration and sec-
ondary succession (Gorchov et al. 1993). For this reason, 
the present study aimed at evaluating the effects of a chronic 
exposure to low, environmentally relevant concentrations of 
endosulfan on blood glucose, glycogen, lipid and protein 
reserves and its bioaccumulation in neotropical bat tissues.
Materials and Methods
The organochlorine insecticide endosulfan (formulation: 
Endosulfan 350 EC Milenia) (6. 7. 8. 9. 10. 10—hexa-chlor—1. 5. 5. 6. 9. 9- hexahydro—6.9—methane- 2.4.3—
benzo (e) dioxatiepin—3- oxide) manufactured by Milenia 
Agro Ciências S.A. was obtained from Federal University 
of Viçosa, Brazil. The adhesive spreader dodecylbenzene 
sulfonic acid, 30 g/L, 3 % m/v is used in association with 
pesticides to increase their efficiency and was added to 
the treatments in the concentration recommended by the 
manufacturer.
Adult male bats (n = 28) (A. lituratus) were captured with 
mist nets around the University campus (20° 45′ S and 42° 52′ 
W) (Viçosa, MG, Brazil). All animals were identified accord-
ing to the key for identification of Brazilian bats (Vizzotto 
and Taddei 1973), weighed, and kept in individual steel cages 
(45 × 22 cm) under natural temperature and light cycles. Cages 
were placed in a fenced-in bat house located under trees at the 
Zoology Museum backyard, at UFV. After 2 days in captiv-
ity, when they were fed papaya (Carica papaya) (200 g) and 
offered water ad libitum, the animals were divided into four 
groups (n = 7), dietary exposed to endosulfan and adhesive 
spreader (AS) for 35 days in one of the following concen-
trations, respectively (g/L): 0.0; 0.0 (Control), 0.0; 0.015 
(AS), 1.05; 0.015 (E1), 2.1; 0.015 (E2). These concentrations 
were chosen because they represent the concentration recom-
mended by the manufacturer for fruit cultures (E1) and twice 
this recommended concentration (E2), reflecting the insecti-
cide levels which bats may find in field crops in this area.
Papayas were used because fruit bats easily accept this 
diet in captivity (Amaral et al. 2012a, b). Fruits were dipped 
in the respective syrup and were hung on an adapted box to 
dry without contact to any surface. Bats were fed daily at 
18h00. Water was available ad libitum. Food consumption 
was monitored daily by placing a known (200 g) amount of 
fruit in each cage. Left overs were collected and weighed 
each morning at 08h00. All captures and experimental 
procedures were performed according with Brazilian laws 
(SISBIO, Process nº 25,048) and the Animal Care and Use 
Committee (CEUA/UFV, Process 69/2014).
At the end of each treatment, animals were euthanized 
and blood was collected in whit heparin tubes. Tissues (liver, 
hind limb, forelimb and breast muscles and adipose tissue) 
1 3
3Bull Environ Contam Toxicol
Our results showed that the HSI was unaltered in exposed 
bats (Table 1), similarly to what was reported in rats treated 
with low doses of endosulfan for 11 weeks (Canlet et al. 
2013). Body weight (BW) also was unaltered following EDS 
exposure (Table 1), unlike the decrease in BW observed for 
bats exposed to higher doses of the organchlorine lindane 
for 3 days (Swanepoel et al. 1999). Plasma glucose levels 
were decreased in E1 exposure treatments relative to con-
trols [F(3.24) = 3.05; p ≤ 0.05] (Fig. 1). Similar results were 
found in female rats exposed to the organochlorine hexa-
chlorobenzene for 3 weeks (Mazzetti et al. 2004). Exposure 
to organochlorines has been shown to induce increases in 
plasma glucose levels in mice (Canlet et al. 2013; Howell 
et al. 2014). One of the possible explanations for the lack 
of hyperglycemia in exposed bats in this study would be 
the lower concentration of endosulfan used, since we aimed 
at simulating the conditions in which bats are exposed in 
nature. Another possibility for the decreased plasma glu-
cose levels in EDS exposed bats would be an impairment of 
the gluconeogenic pathway, as suggested by Mazzetti et al. 
(2004) for rats exposed to hexachlorobenzene.
Glycogen content in the liver and muscle samples showed 
no changes in bats exposed to the insecticide [F(3.24) = 1.89; 
p = 0.16; F(3.24) = 0.81; p = 0.50, respectively] (Table 2). 
Unlike our results, rats treated with hexachlorobenzene 
during 3 weeks showed increased liver glycogen (Mazzetti 
et al. 2004). The longer exposure time we performed here 
likely contributed to the lack of glycogen changes, since the 
dynamics of metabolic homeostasis might have compen-
sated possible changes within the initial days.
Total protein concentration decreased in forelimb muscle 
samples from AS treated bats relative to control exposures 
[F(3.24) = 3.11; p = 0.02] and in hind limb muscle samples 
from E1 (p = 0.003) and AS (p = 0.02) [F(3.24) = 5.91], also 
compared to control. There were no changes in total pro-
tein in the liver [F(3.24) = 1.26; p = 0.31] and breast muscles 
[F(3.24) = 1.37; p = 0.28] (Table 2). A. lituratus exposed to 
the organophophate insecticides spinosyn and fenthion for 
7 and 30 days also did not show changes in total protein 
calculated through the ratio of the areas built in the graph 
(area of the analytic area/internal standard) with known con-
centrations of endosulfan. The area values from the two iso-
mers (α-endosulfan and β-endosulfan) were added and this 
total value was considered the endosulfan total area.
Data was analyzed using GraphPad Prisma statistical soft-
ware (version 5.01). The homogeneity of variance was tested 
by Shapiro–Wilk test followed by a one-way Analysis of 
Variance (ANOVA) and Tukey’s test for multiple compari-
sons among groups. The significance level was set at p < 0.05.
Results and Discussion
Organochlorine exposure has been associated with harmful 
effects on vertebrate metabolism (Thangavel et al. 2010) 
and one of the causes of bat population declines (Gerell and 
Lundeberg 1993; Swanepoel et al. 1999; Clark Jr. 2001).
Table 1 Biological data of Artibeus lituratus dietary exposed to endo-
sulfan and adhesive spreader (AS) in the following concentrations 
(g/L): 0.0; 0.0 (Control). 0.0; 0.015 (AS). 1.05; 0.015 (E1). 2.1; 0.015 
(E2) for 35 days
Experimental groups
ControlAS E1 E2
Body 
weight 
(g)
76.76 ± 2.05 69.96 ± 4.09 74.01 ± 3.97 68.49 ± 2.96
Food con-
sumption 
(g)
40.00 ± 4.86 39.00 ± 4.72 33.00 ± 5.23 39.00 ± 4.41
Liver 
weight 
(mg)
3.64 ± 0.37 3.62 ± 0.39 3.33 ± 0.30 3.19 ± 0.17
Hepato-
somatic 
index
47.33 ± 4.41 48.70 ± 3.85 44.88 ± 3.24 46.87 ± 2.60
Values are expressed as means ± SEM
Fig. 1 Plasma glucose concentration (a) (nmol/L) and carcass fatty 
acids concentration (b) (g/100g) in A. lituratus dietary exposed to 
endosulfan (E) and adhesive spreader (AS), in the following con-
centrations, respectively (g/L): 0.0; 0.0 (Control), 0.0; 0.015 (AS), 
1.05; 0.015 (E1), 2.1; 0.015 (E2) for 35 days. Values are expressed 
as means ± SEM. Asterisk indicates significant difference relative to 
control
 
1 3
4 Bull Environ Contam Toxicol
Abdollahi 2010) though concentrations tested here did not 
show substantial changes considering all analyzed tissues.
In contrast, carcass fatty acid concentrations were 
decreased in both E1 (p = 0.03) and E2 (p = 0.004) exposed 
groups compared to control [F(3.24) = 5.6; p = 0.006] (Fig. 1). 
A mobilization on total fatty acids from peripheral reserves 
was also reported for the same species under an acute expo-
sure to low concentrations of fenthion (Amaral et al. 2012b) 
in the bat P. pipistrellus exposed to the organochlorine lin-
dane for 3 days (Swanepoel et al. 1999). Rezg et al. (2007) 
and Amaral et al. (2012b) also suggest that this mobilization 
might contribute to glucose production via gluconeogenesis 
(glycerol as the precursor) when animals are exposed to pes-
ticides for a short time, which may cause hyperglycemia fol-
lowing a short-term exposure. Given the importance of this 
energy reserve for bats, usually rapidly mobilized during 
fasting for glucose supply to the bloodstream (Freitas et al. 
2006), a decrease of total fatty acids of the carcass due to 
a longer endosulfan exposure might limit bats reproductive 
capacity and their ability to survive food shortages (Moore et 
al. 1984). Furthermore, the results of this study indicate that 
endosulfanmay be considered an energetic stressor, resulting 
reserves in muscles and liver (Amaral et al. 2012a, b). 
Unlike these results, rats treated for 32 days with malathion 
showed decreased liver protein concentration (Rezg et al. 
2007). Karami-Mohajeri and Abdollahi (2010) suggested 
that organochlorine and organophosphosphate pesticides 
might reduce tissue protein content due to glucose produc-
tion from gluconeogenic pathway and/or inhibition of pro-
tein synthesis, though concentrations tested here resulted in 
little changes in protein concentrations.
Overall, total lipid concentrations were increased in AS 
bats liver [F(3.24) = 3.32; p = 0.04] as compared to controls. 
In E1 (p = 0.02) and E2 (p = 0.02), forelimb muscle lip-
ids were decreased as compared to control [F(3.24) = 4.67] 
(Table 2). Hindlimb muscle lipids in E1 (p = 0.003) and AS 
(p = 0.02) were also decreased when compared to control 
[F(3,24) = 5.90]. Lipid contents in adipose tissues showed 
no differences among treatments [F(3.24) = 1.60; p = 0.22]. 
Amaral et al. (2012a) reported decreased muscle lipid 
concentrations in bats exposed to the insecticide spinosad 
for 7 days and increased liver lipid concentrations in bats 
exposed for 30 days. Organochlorines and other pesticides 
may elevate plasma triacylglycerol (Karami-Mohajeri and 
Table 2 Liver and muscle glycogen, lipid and protein contents in Artibeus lituratus dietary exposed to endosulfan and adhesive spreader (AS) in 
the following concentrations, respectively (g/L): 0.0; 0.0 (Control). 0.0; 0.015 (AS). 1.05; 0.015 (E1). 2.1; 0.015 (E2) for 35 days
Metabolic parameters Experimental groups
Control AS E1 E2
Liver glycogen (µmol glicosil-unidades/g)2.17 ± 0.63 4.53 ± 0.70 2.75 ± 0.94 3.46 ± 0.66
Muscle glycogen (µmol glicosil-unidades/g)0.56 ± 0.19 0.84 ± 0.28 0.45 ± 0.16 0.47 ± 0.12
Forelimb muscles protein (g/100g)63.06 ± 5.47 42.19 ± 3.85* 44.87 ± 3.78 52.13 ± 7.24
Hid limb muscles protein (g/100g)57.95 ± 2.11 41.34 ± 4.00* 37.09 ± 2.83* 46.01 ± 5.55
Liver protein (g/100g) 99.13 ± 18.87 68.79 ± 4.73 75.54 ± 7.61 77.81 ± 10.41
Breast muscle protein (g/100g)54.35 ± 5.29 52.12 ± 3.68 56.13 ± 4.52 59.51 ± 5.09
Liver lipids (g/100g) 6.90 ± 0.70 9.87 ± 0.77* 8.37 ± 0.82 7.23 ± 0.60
Forelimb muscles lipids (g/100g)3.52 ± 0.31 4.61 ± 0.52 5.27 ± 0.25* 5.34 ± 0.44 *
Hind limb muscles lipids (g/100g)5.06 ± 0.40 5.55 ± 0.47 5.05 ± 0.46 4.97 ± 0.62
Breast muscles lipids (g/100g)6.90 ± 0.73 8.27 ± 0.97 7.23 ± 0.57 6.29 ± 0.29
Adipose tissue lipids (g/100g)51.56 ± 2.65 59.06 ± 3.55 53.70 ± 3.58 61.75 ± 4.63
*Significant difference relative to control
Table 3 Endosulfan concentraction found in the diet, liver and adipose tissue of Artibeus lituratus dietary exposed to endosulfan and adhesive 
spreader (AS) in the following concentrations, respectively (g/L): 0.0; 0.0 (Control). 0.0; 0.015 (AS). 1.05; 0.015 (E1). 2.1; 0.015 (E2) for 35 days
Endosulfan concentraction
Liver (ng/g) Adipose tissue (ng/g)Diet (mg/Kg)
Control ND ND ND
AS ND ND ND
E1 0.92 ± 0.00 3.08 ± 0.03 57.87 ± 3.25
E2 6.48 ± 0.00 13.68 ± 0.00 61.47 ± 5.78
ND non-detectable
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5Bull Environ Contam Toxicol
Acknowledgments We thank the National Counsel of Technologi-
cal and Scientific Development (CNPq, Brazil) for supporting this 
research.
References
Agbohessi TP, Toko II, N’tcha I, Geay F, Mandiki SNM, Kestemont 
P (2014) Exposure to agricultural pesticides impairs growth, feed 
utilization and energy budget in African Catfish Clarias gariepi-
nus (Burchell, 1822) fingerlings. Int Aquat Res 6:229–243
Allinson G, Mispagel C, Kajiwara N, Anan Y, Hashimoto J, Lauren-
son L, Allinson M, Tanabe S (2006) Organochlorine and trace 
metal residues in adult southern bent-wing bat (Miniopterus 
schreibersii bassanii) in southeastern Australia. Chemosphere 
64:1464–1471
Amaral TS, Carvalho TF, Barros MS, Picanço MC, Neves CA, Freitas 
MB (2012a) Short-term effects of spinosyn’s family insecticide 
on energy metabolism and liver morphology in frugivoros bats 
Artibeus lituratus (Olfers. 1818). Braz J of Biol 72(2):299–304
Amaral TS, Carvalho TF, Silva MC, Goulart LS, Barros MS, Picanço 
MC, Neves CA, Freitas MB (2012b) Metabolic and histopath-
ological alterations in the fruit-eating bat Artibeus lituratus 
induced by the organophosphorous pesticide fenthion. Acta Chi-
ropterol 14(1):225–232
Bayata S, Geiser F, Kristiansen P, Wilson SC (2014) Organic contami-
nants in bats: trends and new issues. Environ Int 63:40–52
Bennett BS, Thies ML (2007) Organochlorine pesticide residues 
in guano of Brazilian freetailed bats, Tadarida brasiliensis 
Saint-Hilaire, from east Texas. Bull Environ Contam Toxicol 
78:191–194
Canlet C, Tremblay-Franco M, Gautier R, Molina J, Métais B, Blas-Y 
Estrada F, Gamet-Payrastre L (2013) Specific metabolic finger-
print of a dietary exposure to a very low dose of endosulfan. J 
Toxicol 2013:545802
Carneiro FF, Rigotto RM, Augusto LGDS, Friedrich K, Burigo AC 
(2015) Dossiê ABRASCO: um alerta sobre os impactos dos 
agrotóxicos na saúde.
Clark Jr. DR (2001) DDT and the decline of free-tailed bats (Tadarida 
brasiliensis) at Carlsbad Cavern, New Mexico. Arch Environ 
Contam Toxicol 40:537–543
Coats JR (1990) Mechanisms of toxic action and structure–activity 
relationships for organochlorine and synthetic pyrethroid insec-
ticides. Environ Health Perspect 87:255–262
Dennis GC, Gartrell BD (2015) Nontarget mortality of New Zealand 
lesser short-tailed bats (Mystacina tuberculata) caused by diphac-
inone. J Wildl Dis 51:177–186
Folch J, Less M, Slorne SGHA (1957) A simple method for the iso-
lation and purification of total lipids from animal tissue. J Biol 
Chem 226:226–497
Freitas MB, Welker AF, Pinheiro EC (2006) Seasonal variation and 
food deprivation in common vampire bats (Chiroptera: Phyllos-
tomidae). Braz J Biol 66:1051–1055
Gerell R, Lundeberg KG (1993) Decline of a bat Pipistrellus pipistrel-
lus population in an industrialized area in south Sweden. Biol 
Conserv 65:153–157
Gorchov DL, Cornejo F, Ascorra C, Jaramillo M (1993) The role of 
seed dispersal in the natural regeneration of rain forest after strip-
cutting in the Peruvian Amazon. In: Fleming TH and Estrada A 
(eds). Frugivory and seeddispersal: ecological and evolutionary 
aspects. W. Kluwer Academic Publishers, Dordrecht, pp 339–349
Howell GE, Meek E, Kilic J, Mohns M, Mulligan C, Chambers JE 
(2014) Exposure to p, p’-dichlorodiphenyldichloroethylene 
(DDE) induces fasting hyperglycemia without insulin resistance 
in male C57BL/6 H mice. Toxicology 5:6–14
in decreased lipid reserves. This is consistent with the accu-
mulation of organochlorine in the tissues from bats that 
increase the metabolic rate and consequently the metaboliz-
ing lipids. Taken together, these alterations might have con-
sequences for bats, as the decline in energy reserves would 
force an increase in foraging time, increasing the risk of pre-
dation or accident (Allinson et al. 2006; Bayata et al. 2014).
Another concern we wanted to address was to determine 
whether or not the 35-days exposure to environmentally 
relevant concentrations of endosulfan would cause bio-
accumulation in bat tissues. Residues were identified by 
comparing the compound retention time to the pattern for 
this chemical. The peaks with retention time (RT) of 7.629 
and 8.878 correspond to α-endosulfan and β-endosulfan. 
The 6.456 peak correspond to the internal standard (para-
thion). Organochlorine in tissue residues have been reported 
in several temperate insectivorous bat species (Gerell and 
Lundberg 1993; Kannan et al. 2010; Stechert et al. 2014). 
Though to our knowledge, this is the first time that endo-
sulfan bioaccumulation is being reported for a Neotropical 
fruit-eating species. EDS concentrations detected in bat tis-
sues (Table 3) were lower than what was reportedfor oral 
acute doses (LD50: 70 mg/kg) in rats (Tomlin 2006).
Toxic effects caused by organochlorine accumulation 
in body lipids happen through lipids mobilization. Bioac-
cumulation of pesticides in bat tissue leads to decreased 
antioxidant capacity (Oliveira 2013; Naidoo et al. 2015) 
and decreased activity of the complement system, resulting 
in damage to the innate immunity (Lilley et al. 2013). The 
decreased immune response can increase bat’s vulnerability 
to diseases like white nose syndrome, leading to population 
declines (Bennett and Thies 2007; Kannan et al. 2010), as 
proven for Mystacine tuberculata exposed to the rodenticide 
diphacinone in New Zealand (Dennis and Gartrell 2015) 
and seem to be the case for Tadarida brasiliensis mexicana 
in New Mexico, USA (Clark Jr. 2001).
In summary, our results show that a 35-days dietary 
exposure to low, environmentally relevant concentrations 
of the endosulfan formulation tested affected carcass and 
muscle lipid energy reserves in the neotropical fruit bat A. 
lituratus. The decreased carcass fatty acids reported here, 
which in E2 bats was about half the concentration found 
in controls, could be critical for energy supply during high 
energy demand periods, such as reproduction and seasonal 
food shortages. This study demonstrated changes in fruit 
bat energy reserves and bioaccumulation following EDS 
exposure. Such bioaccumulation and physiological changes 
represent additional threats to the long-term viability of 
neotropical fruit bat population, which are already at risk 
due to forest fragmentation and habitat loss. Further, A. litu-
ratus is an important seed disperser in tropical forests, thus 
long term health threats to this species may complicate con-
servation efforts.
1 3
6 Bull Environ Contam Toxicol
Paulino MG, Benze TP, Sadauskas-Henrique H, Sakuragui MM, Fer-
nandes JB, Fernandes MN (2014) The impact of organochlorines 
and metals on wild fish living in a tropical hydroelectric reservoir: 
bioaccumulation and histopathological biomarkers. Sci Total 
Environ 497–498:293–306
Rezg R, Mornagui B, El-Fazaa S, Gharbi N (2007) Biochemical evalu-
ation of hepatic damage in subchronic exposure to malathion in 
rats: effect on superoxide dismutase and catalase activities using 
native PAGE. C R Biol 331:655–662
Saiyed H, Dewan A, Bhatnagar V, Shenoy U, Shenoy R, Rajmohan H, 
Patel K, Kashyap R, Kushyap R, Kulkarni P, Rajan B, Lakkad B 
(2003) Effect of Endosulfan on male reproductive development. 
Environ Health Perspect 111:1958–1962
Singh SK, Pandey RS (1990) Effect of sub-chronic endosulfan expo-
sures on plasma gonadotrophins, testosterone, testicular testos-
terone and enzymes of androgen biosynthesis in rat. Indian J Exp 
Biol 10:953–956
Sjörgren B, Noerdenskjöld T, Holmgeen H, Mollerstrom J (1938) 
Beitrag zur Kenntnis der Leberrhythmik (glycogen, Phosphor und 
Calcium in der Kaninchenleber). Pflugers Arch EJP 240:427–448
Stechert C, Kolb M, Bahadir M, Djossa BA, Fahr B (2014) Insecticide 
residues in bats along a land use-gradient dominated by cotton 
cultivation in northern Benin, West Africa. Environ Sci Pollut Res 
21:8812–8821
Swanepoel RE, Racey PA, Shore RF, Speakman JR (1999) Energetic 
effects of sublethal exposure to lindane on pipistrelle bats (Pip-
istrellus pipistrellus). Environ Pollut 104:169–177
Thangavel P, Sumathiral K, Maheswari S, Rita S, Ramaswamy M 
(2010) Hormone profile of an edible, freshwater teleost, Sarother-
odon mossambicus (Peters) under endosulfan toxicity. Pestic Bio-
chem Physiol 97:229–234
Tomlin CDS (ed) (2006) A world compendium: the pesticide manual, 
14th edn. BCPC, Hampshire
Van Dyk JS, Pletschke B (2011) Review on the use of enzymes for 
the detection of organochlorine, organophosphate and carbamate 
pesticides in the environment. Chemosphere 82:291–307
Vizzotto LD, Taddei VA (1973) Chave para determinação de 
quirópteros brasileiros. Universidade Estadual Paulista, São José 
do Rio Preto.
Kalender S, Kalender Y, Ogutcu A, Uzunhisarcikli M, Durak D, Açik-
goz F (2004) Endosulfan-induced cardiotoxicity and free radical 
metabolism in rats: the protective effect of vitamin E. Toxicology 
202:227–235
Kannan K, Yun SH, Rudd RJ, Behr M (2010) High concentrations of 
persistent organic pollutants including PCBs, DDT, PBDEs and 
PFOS in little brown bats with white-nose syndrome in New 
York. USA. Chemosphere 80:613–618
Karami-Mohajeri S, Abdollahi M (2010) Toxic influence of organo-
phosphate. carbamate. and organochlorine pesticides on cellular 
metabolism of lipids, proteins, and carbohydrates: a systematic 
review. Hum Exp Toxicol 30:1119–1140
Kuvarega AT, Taru P (2007) Accumulation of endosulfan in wild rat, 
Rattus norvegious as a result of application to soya bean in Mazoe 
(Zimbabwe). Environ Monit Assess 124:333–345
Lilley TM, Ruokolainen L, Meierjohann A, Kanerva M, Stauffe J, 
Laine VN, Atosuo J, Lilius EM, NIkinmaa (2013) Resistance 
to oxidative damage but not immunosuppression by organic tin 
compounds in natural populations of Daubenton’s bats (Myotis 
daubentonii). Comp Biochem Phys C 3:298–305
Mazzetti MB, Taira MC, Lelli SM, Dascal E, Basabe JC, de Viale LC 
(2004) H exachlorobenzene impairs glucose metabolism in a rat 
modelo f porphyria cutanea tarda: a mechanistic approach. Arch 
Toxicol 78:25–33
Moore BJ, Olsen JL, Marks F, Brasel JA (1984) The effects of high fat 
feeding during one cycle of reproduction consisting of pregnancy, 
lactation and recovery on body composition and fat pad cellular-
ity in the rat. J Nutr 114:1566–1573
Naidoo S, Vosloo D, Schoeman MC (2015) Haematological and 
genotoxic responses in an urban adapter, the banana bat, forag-
ing at wastewater treatment works. Ecotoxicol Environ Safe 114: 
304–311
O’shea TJ, Everette AL, Ellison LE (2001) Cyclodiene insecticide 
DDE, DDT, Arsenic, and mercury contamination of big brown 
bats (Eptesicus fuscus) foraging at a Colorado Superfund site. 
Arch Environ Contam Toxicol 40:112–120
Oliveira JM (2013) Exposição Crônica a Baixas Concentrações do 
Inseticida Endosulfan Altera a Capacidade Antioxidante de 
Morcegos Frugívoros (Artibeus lituratus, OLFERS, 1818). Dis-
sertation, Federal University of Viçosa.
1 3
	Low, Chronic Exposure to Endosulfan Induces Bioaccumulation and Decreased Carcass Total Fatty Acids in Neotropical Fruit Bats
	Abstract
	Materials and Methods
	Results and Discussion
	References