In water endosulIan

has a halI liIe oI 35 to 150 days

The halI liIe oI endosulIan
varies Irom 60 days (alpha- endosulIan) to 800 days
(beta-endosulIan), It does not easily
dissolve in water and may accumulate in bodies oI
Iish and other aquatic organisms30,69.


endosulfan endosulfan


2.




2.2. Persistence.
EndosulIan is recognized as a Persistent Toxic Substance (PTS) by the United Nations
Environment Programme (UNEP, 2003). In soil,
but is rapidly degraded in water (ATSDR, 2000).
2.3. ioaccumulation.
The maximum bioconcentration Iactors () oI endosulIan in aquatic systems are
usually less than 3,000, and metabolism studies with laboratory animals indicate that
endosulIan does not bioconcentrate in Iatty tissues and milk (ATSDR, 2000), suggesting that
it has only a moderate potential Ior bioaccumulation.
3. Potential Ior human exposure to endosulIan.
3
EndosulIan contamination is Iound throughout the environment and is present in many
superIund sites in the United States (Kullman and Matsumura, 1996; ATSDR, 2000). A
review oI commonly used pesticides in anadian air claimed that endosulIan is one oI the
most abundant and ubiquitous organochlorine pesticides in the North American atmosphere
today (Tuduri et al, 2006). A study in Denmark on pesticide residues (including endosulIan)
in apples reported mean concentrations oI 0.482, 0.468, and 0.024 mg/kg Ior endosulIa-0t, f3,
and sulIate, respectively (Rasmusssen et al., 2003). The study added that simple washing
does not signiIicantly decrease the amount oI pesticides and that endosulIan sulIate residues
were increased by 34 during long-term storage oI apples.
Humans can be exposed to endosulIan percutaneously or through ingestion and inhalation
(Kalender et al., 2004). or individuals in occupational settings or people living near
hazardous waste sites or agricultural Iields applied with endosulIan, the primary routes oI
exposure would be inhalation and dermal exposure. In a study oI spraymen's exposure to
endosulIan Irom application oI this pesticide on Iruit orchards, a mean dermal exposure 24.7
mg/hr (range oI 0.6-95.3 mg/hr) and a mean inhalation exposure oI 0.02 ng/hr (range oI
0.01-0.05 mg/hr) were estimated (WolIe et al., 1972). In another study oI women living in
Southern Spain, where large amounts oI endosulIan were used in intensive greenhouse
agriculture, endosulIan residues were detected in breast milk (11.38 mg/mL) and adipose
tissues (17.72 ng/g) oI these women (errillo et al., 2005). errillo et al Iurther argued that
their Iindings support the lipophilicity oI endosulIan and its elimination by milk secretion.
EndosulIan and metabolites were also detected in Iats oI 30-40 oI children hospitalized
aIter exposure to endosulIan in agricultural regions oI Spain (Olea et al., 1999).
4
or the general population, the main route oI exposure to endosulIan is ingestion oI Iood
or tobacco products containing residues oI endosulIan. In 2001, the Total Diet Study
conducted by the DA's pesticide residue monitoring program determined endosulIan as one
oI the 19 most Irequently Iound residues (those Iound in ~2 oI the samples), endosulIan as
one oI the 5 most Irequently observed chemicals, and endosulIan with an 18 percent
occurrence in the Iour market baskets analyzed (1030 Iood items). Albeit, the levels oI
endosulIan residues, were well below regulatory limits (DA, 2001).
3.1. Release oI endosulIan to the environment.
The major source oI endosulIan distribution to the environment is spray driIt Irom aerial
applications oI this pesticide on crops. However, endosulIan can also be distributed through
run-oII water Irom Iields sprayed with this pesticide (Kennedy et al, 2001) and through
volatilization where it can undergo long-range atmospheric transport, by which traces oI this
pesticide have been Iound in the Arctic (Jantunen and idleman, 1998). EndosulIan has been
detected in air, surIace water, groundwater, soil, and sediment (ATSDR, 2000).
3.2. Environmental Iate.
EndosulIan is not very soluble in water. The log Kow (Octanol-Water partition
coeIIicient - describes the partitioning oI organic pollutants between water phase and
octanol) oI endosulIan-a and 03a re 3.83 and 3.52, respectively (Hansch et al., 1995).
EndosulIan has been Iound at low concentrations in Iew surIace water and groundwater
samples at hazardous waste sites (ATSDR, 2000).
EndosulIan partitions to the atmosphere and soils or sediments (ATSDR, 2000). The
Henry's law constant (describes partitioning oI organic pollutants between the water phase
and air in the environment) Ior endosulIan-cL and 03a re 0.7-12.9 and 0.04-2.12 Pa m3 mol1 at
5
20', respectively (German ederal Environment Agency, 2004). These values indicate that
the at-isomer is expected to be more mobile in the atmosphere because it is more volatile than
the 03is omer. This is supported by a review on currently used pesticides in anadian air
which stated that endosulIan-03 is always detected in lesser amounts than the a-isomer in the
atmosphere, partly due to the composition oI technical-endosulIan (contains more at-isomer)
but also because the a-isomer is more volatile (Tuduri et al., 2006).
Results oI several studies indicate that both isomers oI endosulIan strongly adsorb to soil.
The Koc (organic carbon-water partition coeIIicient - describes partitioning oI organic
pollutants between water phase and natural organic matter in soils or sediments) oI
endosulIan-at and 03a re estimated to be 2,887 and 1,958, respectively, suggesting that
mobility oI endosulIan in soil or sediment is expected to be slight (ATSDR, 2000).
EndosulIan does not bioaccumulate to high concentrations in terrestrial or aquatic
organisms. Maximum bioconcentration Iactors are less than 3,000, and residues are
eliminated Irom organisms very rapidly (ATSDR, 2000).
3.3. Degradation in the environment.
EndosulIan-03 is slowly converted to the more stable at-isomer at high temperatures (Rice
et al., 1997). EndosulIan is degraded to endosulIan diol by hydrolysis in surIace water and
groundwater. In sediment and soil, endosulIan is degraded primarily to endosulIan sulIate by
Iungal metabolism and to endosulIan diol by bacterial metabolism. iotransIormation oI
endosulIan in soil under aerobic conditions yield endosulIan sulIate, endosulIan diol, and
endosulIan lactone. In anaerobic conditions, the metabolites endosulIan diol, endosulIan
sulIate, and endosulIan hydroxyether are Iormed (ATSDR, 2000).
3.4 Metabolism in mammals.
6
In animals, endosulIan-a or 03c an be converted to endosulIan sulIate or endosulIan diol
(WHO, 1984), which can be Iurther metabolized to endosulIan lactone, hydroxyether, and
ether (ATSDR, 2000). In humans, the metabolites endosulIan sulIate, alcohol, ether, and
lactone were detected in urine (Martinez Vidal et al., 1998; ATSDR, 2000) and serum
(Arrebola et al., 2001).
Although the metabolites diol, ether, and lactone are non-toxic, the metabolite endosulIan
sulIate is nearly as toxic as the parent endosulIan, does not undergo Iurther degradation, and
residues tend to increase in the environment (Kennedy et al., 2001).
Why study endosulIan?
Endocrine disruption seen in children in developing countries living nearby Iields sprayed
with endosulIan could have been caused by chronic exposure to this pesticide. In the US, a
concern Ior chronic exposure oI children to endosulIan should be raised Ior the Iollowing
reasons: 1) endosulIan is a pesticide still commonly applied to Iruits and vegetables and its
residues are detected in a number oI Iood items Irom supermarkets, albeit at levels below
9
regulatory limits; 2) endosulIan is a common contaminant Iound in the environment; 3)
endosulIan and its metabolite endosulIan sulIate are persistent substances; and 4) children are
particularly susceptible to the toxicity oI endosulIan (ATSDR, 2000).
The results oI the Saiyed et al, 2003 epidemiological study, where testosterone levels were
decreased and delayed male sexual maturity were seen in children who were chronically
exposed to endosulIan, coupled with evideIice that organochorines induce P enzymes
(Siddiqui et al., 1987), led us to think that a very likely mechanism Ior endosulIan's
endocrine disruption is through its interaction with !# and/or AR and induction oI the
P enzymes.
This research consisted oI two major areas oI investigation. irst, the human metabolic
pathway oI endosulIan was determined and is discussed in hapter 1. Second, these studies
characterized the role oI endosulIan in inducing P450s as a possible mechanism oI endocrine
disruption and are discussed in hapter 2.
10

@e Lwo reporLed mecanlsms of neuroLoxlclLy
lnclude 1) blockage of neuroLransmlLLer recepLors (Zaldl eL al 1983 Aballs eL al 1986) and
2) lnLerference wlL synLesls degradaLlon and release/reupLake of neuroLransmlLLers (aul
eL al 1994) CA8AanLagonlsm ls Le wldely accepLed mecanlsm of acuLe or neuronLoxlclLy
(A@ 2000)
ln Le u endosulfan was llnked Lo Le deaL of one farmer and permanenL neurologlcal
lmpalrmenL of anoLer (8randL eL al 2001) ln Cuba 13 people dled of endosulfan
polsonlng ln Le provlnce of MaLanzas were a LoLal of 63 people became lll afLer eaLlng
food conLamlnaLed wlL endosulfan (Campalgner Aug 1999) ln 8orgou 8enln endosulfan
caused 37 deaLs durlng Le 19992000 coLLon season (@on eL al 2000) ln Le same reglon
a boy dled ln 1999 afLer lngesLlng corn conLamlnaLed wlL endosulfan (Myers 2000) ln
2
udan ln 1988 endosulfan barrels wased ln lrrlgaLlon canals caused fls kllls and deaL of
Lree people wo drank waLer from Le canal (lnam 1993) ln 1993 ln Columbla 60
polsonlngs and 1 deaL occurred due Lo endosulfan use on coffee (Anu 1994)
ln laboraLory anlmals endosulfan produces neuroLoxlclLy effecLs wlc are belleved Lo resulL from over
sLlmulaLlon of Le cenLral nervous sysLem Neurotox|c|ty |s the ma[or end po|nt of concern |n acute
endosu|fan exposure |n human be|ngs and exper|menta| an|ma|s ndosu|fan |s neurotox|n caus|ng
convu|s|ons and death ndosu|fan |s part|cu|ar|y neurotox|c for both |nsects and mamma|s |nc|ud|ng
humans It is an antagonist oI the chain oI gamma-aminobutyric acid (GAA) receptors in the brain,
reducing the uptake oI chloride ions by neurons, which results in uncontrolled excitation (UNEP/AO,
2007). It also inhibits calcium and magnesium uptake, and the enzyme ATPase. oth enzymes are
involved in the transIer oI nerve impulses. Among the most characteristic symptoms oI poisoning by
endosulIan are hyperactivity, tremors, convulsions, lack oI coordination, dizziness, diIIiculty breathing,
nausea and vomiting, diarrhoea, and in severe cases, impaired consciousness (ATSDR, 2000). Doses as
low as 35 mg/kg have caused human death (IPS, 2000). Neurotoxicity is oI major concern with acute
endosulIan exposure oI human beings and animals. EndosulIan is a neurotoxic organochlorine insecticide
oI the
cyclodiene Iamily oI pesticides. It is an endocrine disruptor, and it is
highly acutely toxic. Doses as low as 35 mg/kg have been
documented to cause death in humans, and many cases oI sub-lethal
poisoning have resulted in permanent brain damage.
). EndosulIan is known to disrupt
neurotransmitter transmission in cholinergic neurotransmitter systems (systems which use
acetylcholine as a neurotransmitter), altering the concentrations oI melatonin,
acetylcholine, serotonin, dopamine and norepinephrine (Goulet & Hontela 2003; Harris et
al. 2000; Park et al. 2001; Sparling et al. 2001). There are a number oI hypothesized
biochemical pathways which are targeted by endosulIan which would lead to
melanophore aggregation, and at present the aIIected pathway(s) oI the aIIlicted P.
regilla tadpoles oI this experiment is not known. EndosulIan is known to disrupt neurotransmitter
transmission in cholinergic
neurotransmitter systems (systems which use acetylcholine as a neurotransmitter),
altering the concentrations oI melatonin, acetylcholine, serotonin, dopamine and
norepinephrine (Goulet & Hontela 2003; Harris et al. 2000; Park et al. 2001; Sparling et
al. 2001). There are a number oI hypothesized biochemical pathways which are targeted
by endosulIan, which would lead to melanophore aggregation. atecholamines and
melatonin have shown to increase in animals exposed to endosulIan due to endosulIan
stimulating the sympathetic nervous system (Park et al. 2001; Rohr et al. 2003; Scholz &
Hopkins 2006). These neurotransmitters act as antagonists to MSH, and Iavor a decrease
in melanophoric cAMP levels, and thus Iacilitate melanophore aggregation (Aspengren et
al. 2003; Hadley & Goldman 1969; Novales & Davis 1969; White et al. 1987). The alpha isomer is more
neuro-toxic and its acute
toxicity against mammals is more than three times
that oI the beta-isomer109


There is experimental evidence oI adverse
eIIects oI endosulIan on the male reproductive
system, delaying sexual maturity and interIering with
the sex-hormone synthesis118. EndosulIan is a
proven endocrine disruptor6,9. It has potential to
induce hypo thyroidism66. Long term health eIIects
are not properly studied, experimented or
documented world wide.
EndosulIan exhibits estrogenic properties9,43,
comparable to that oI DDT9. It competes Ior estradiol
Ior binding to estrogen receptors, thereby inhibiting
hormonal Iunction 107. The estrogenic potential oI endosulIan increases in the presence oI other
estrogenic organochlorines 110 .It induces proliIeration oI human breast estrogen sensitive M7 cells49,
(invitro) thereby increasing breast cancer risk117. It harms the reproductive system by aIIecting semen
quality, sperm count, spermatogonial cells, sperm morphology and other deIects in male sex
hormones11.EndosulIan |is having| have the capacity to alter the genetic material particularly
chromosomes in mammalian cultures119. It is Iound to inhibit testicular androgen biosynthesis in lab
animal experiments30 and exhibits signiIicant risk in renal and testicular damage. It may have adverse
eIIects on central nervous system by inhibiting brain acetyl cholinesterase16, causing uncontrolled
discharge oI acetyl choline. EndosulIan ingestion is known to
aIIect the kidneys and liver50. It inhibits leucocyte
and macrophage migration (this is the inhibition oI the
natural immune system by disrupting anti-body
protection) causing adverse eIIects on humoral and
cell-mediated immune system30. It is also a potential
tumor promoter67.
Many studies related to its acute and chronic
toxicity in laboratory animals are available.
EndosulIan is highly toxic to rats and mice13, 30.Some
studies suggests its teratogenic28 and carcinogenic
properties21 on rats and mice. It directly aIIects the
central nervous system, causes liver and kidney
(chronic glumerulonephrosis) damage6 in rats and
mice. It also impairs the reproductory system oI
rats39. ehavioural and neurological changes have
also been noticed30. Thyroid Iollicular damage in
mouse has been reported30,66. EndosulIan is known
to damage the endocrine system, nervous system,
circulatory, reproductory, respiratory and excretory
systems and developing Ioetus.6,7,14,15,16,21,30,37
The National Institute oI Occupational Health
(India) have linked the higher prevalence oI
neurobehavioral disorders, congenital malIormations
in Iemale children and abnormalities related to male
reproductive systems to the continuous exposure to
endosulIan spray. The study was conducted among
children in one oI the villages in Kasaragod District
(in the South Indian State oI Kerala) where
endosulIan was aerially sprayed64. EndosulIan is
implicated in the occurrence oI adverse health eIIects
particularly in rural communities in South East Asia,
Southern PaciIic and Sub-Saharan AIrica.69,71
hronic eIIects reported in humans include birth deIects, congenital reproductive
disorders, long-term brain damage, recurrent convulsions, epilepsy, autism, delayed
sexual maturity, endometriosis, menstrual disorders, early menarche, male breast
enlargement, various cancers, congenital intellectual disability, cerebral palsy,
psychiatric disturbances, and vision impairment and loss (Aleksandrowicz 1979;
Pradhan et al 1997; Quijano 2002; NIOH 2003; Saiyed et al 2003; Roberts et al
2007; Venugopal 2008).
# TTY
Exposure through certain conditions oI use (e.g. lack oI protective equipment), and bystander`
exposure have been linked to congenital physical disorders, mental retardations and deaths in Iarm
workers and villagers in developing countries.14 The sub acute and chronic toxicity studies oI
endosulIan in animals suggest that the liver, kidneys, immune system, and testes are the main target
organs. A number oI neurotoxicity studies were available, primarily in the rat. An acute
neurotoxicity study in rats (gavages) showed a greater sensitivity oI Iemales compared to males 15
Long term exposure is linked to immunosuppression, neurological disorders, congenital birth
deIects, chromosomal abnormalities, mental retardation, impaired learning and memory loss. 16
Long-term oral and dermal exposure in rats has been Iound to result in rapidly progressive
Glomerulonephritis. This is a renal disease that aIIects the small blood vessels in the kidneys. Longterm
oral and dermal exposure in male rats has been observed to cause aneurysms (blood-vessel
dilations which iI ruptured can even lead to death).17 onsumption oI endosulIan at low dose Ior
longer duration can ultimately lead to diIIerential alterations oI Monoamines (an important class oI
neurotransmitters) in various regions oI the rat central nervous system.18
There is some indication that endosulIan can have adverse eIIects on the immune system at low
levels oI exposure.19 There is mounting evidence that organochlorine compounds can act as
hormones. These compounds, including DDT, Ps, and endosulIan, may also be part oI the cause
Ior the disease in the quality oI semen, an increase in testicular and prostate cancer, an increase in
deIects in male sex organs, and increases incidence oI breast cancer which has been observed in the
last IiIty years. 20

The brain is thought to
be susceptible to lipid peroxidation and oxidative injury,
since it contains membranes rich in polyunsaturated Iatty
acids. It is antioxidant-poor compared to other organs and has a high content oI iron. Jayaraman,
2008
Since the brain makes up only 2% of the total body
weight of a human, yet consumes approximately 20% of
the available oxygen, it is an excellent environment for
the occurrence of oxidative stress [73]. The brain also
contains high levels of lipids while possessing low amounts
of antioxidants, thus further increasing its susceptibility to
damage as the result of ROS and oxidative stress [74 (Katie, 2011).
The brain is the most susceptible organ to oxidative
damage due to its high oxygen demand (1,2). Elevated oxygen consumption may
lead to oxidative stress. The pathology of aging-associated neurodegeneration is
increasingly linked to oxidative and nitrosative stress mediated by free radicals
(3,4). Within the cell, free radicals, such as superoxide (O2
-) and hydroxyl (OH),
are normal products of cellular oxygen metabolism. Moreover, other molecules
like H2O2 and peroxynitrite (ONOO-), although not themselves free radicals, can
lead to generation of free radicals. Together, these highly reactive metabolites are
referred to as reactive oxygen (ROS) and nitrogen species (RNS). Oxidative
stress arises from an imbalance between cellular ROS/RNS production and the
ability of cells to defend themselves against this stress. The attack of free radicals,
ROS and RNS causes cellular damage by oxidizing proteins, membrane lipids,
and DNA. (SCHMTT-SCHLLG 2005,).
Oxygen Iree radicals have been implicated in the
development oI many neuronal disorders and
brain dysIunction.'-s1 The brain appears to be
particularly vulnerable to oxidative damage since
it contains relatively high concentrations oI readily
peroxidizable Iatty acids.|9r10V1a rious brain
regions are highly enriched in which
can catalyze the production oI damaging oxygen
Iree radical species. Moreover, the brain, which
consumes a signiIicant Iraction oI the total
oxygen demand oI the body,|161is rather poorly
endowed with protective antioxidant enzymes
or antioxidant compounds.7r181T hus, the susceptibility
oI the brain to oxidative stress
might account Ior an eIIect peculiar to this tissue
in vitro. ### 0t nl.1998

@e braln ls deflclenL ln oxldaLlve defense mecanlsms and
ence ls aL greaLer rlsk of damage medlaLed by reacLlve oxygen specles
() resulLlng ln molecular and cellular dysfuncLlonWy braln ls more vulnerable Lo oxldaLlve sLress ?
@e cenLral nervous sysLem (Cn) ls
especlally vulnerable Lo free radlcal damage
because of braln's lg oxygen consumpLlon
lLs abundanL llpld conLenL and Le relaLlve
pauclLy of anLloxldanL enzymes as compared
wlL oLer Llssues (3) Moreover braln as
a lg raLlo of membrane surface area Lo
cyLoplasmlc raLlo exLended axonal
morpology prone Lo ln[ury and neuronal
cells are nonrepllcaLlng can lncrease
Le permeablllLy of Le blood braln barrler
alLer Lubulln formaLlon and lnlblL Le
mlLocondrlal resplraLlon lf uncecked lL
can lead Lo a geomeLrlcally progresslng llpld
peroxldaLlon Lvldence also lndlcaLe LaL
may sLlmulaLe exLracellular release of
exclLaLory amlnoaclds (4) CluLamaLe ls Le
ma[or exclLaLory amlnoacld ln Le braln lL
acLs Lroug varlous Lypes of lonoLroplc
recepLors Le mosL slgnlflcanL belng nMA
recepLors @ere seems Lo be a bldlrecLlonal relaLlonslp beLween Le producLlon
and release of exclLaLory amlnoaclds (3)
lree radlcals generaLed ln Le braln are also
reporLed Lo lnfluence gene expresslon
subsequenLly effecLlng apopLosls and
neuronal deaL (6) CupLa eL al 2003