In: Pollution Control ISBN: 978-1-61668-584-3

Editor: H.R. Velasquez, pp. 43-82 © 2011 Nova Science Publishers, Inc.
Brooke, D., de
G., Koch, V.,
n union system
r.Jhere, 59, 473-

, L., Rappe, C.,

ls in household Chapter 2
roxicology and

: of manure and
f Chenopidium
mrosphere, 57, CONTAMINATION? NATURALAND ANTHROPIC
STRESSORS ON FRESHWATER DECAPOD
and phytotoxic
icultural uses. CRUSTACEANS
involved in the
·-sea interface.
P. Collins *, V. Williner and M. Montagna
Instituto Nacional de Limnología, Ciudad Universitaria Pje El Pozo s/n 3000
, R.D., Scoffin,
mts in sewage Santa Fe, Argentina
non-analytical

of potentially-
Abstract
'on Society, 62,
Organisms and populations in a freshwater ecosystem respond, when the
environment is affected to extreme situations, of different mode. These stressors
Cd in a small could be the product of human activities (e.g. farmland, industry, cities) or
alterations of natural cycle (e.g. abnormal drought and flooding; or extreme
·trace elements maximum and mínimum temperature). According to the intensity (temporal,
spatial and amount of agent), the response vary since behavioral aspect to
~~ality, 27, 715-
survival diminution. Biological communities are in equilibrium with all their
components. However, this stability could crack, when their members change
iactionation of their relative relationship, or when new elements are incorporated; or the main
wo composts to cycles are modified. These elements can affect the interna! biochemical
composition; frequency and alteration in cell of sorne organs leading to death,
disease, reproduction failures or diminished growth._Among freshwater
Copper and Zn crustaceans, crabs and prawns have been known as sensitive to environmental
f livestock and stress; and their biological characteristics allow us to use in them ecological and
toxicological studies. Moreover, the climatic changes together to quantitatively
and variety increase of products that man had produced, used and flushes in the

' E-mail address: [email protected]

44 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

environment, provokes constant risk to the fauna and thus creates the necessity of
C::2~ C..:~~ rain
constant update studies._The continental aquatic environments, by its relative
instability, and proximity to different human activities (industrial, farmland, and ••• • • '•'',. irrigation
city) are more frequently affected by climatic phenomenon and xenobiotic cities
products. When the actions reach the lotic and lentic environments interacts with
each member of the communities. The aim of this chapter is analyze and show fannland ·
the effects that it be observed in freshwater decapods dueto natural and anthropic
stressors. The identification of the process that occurs in the environment is very
important, indicating when the species are affected by natural or anthropic superficial and
stressors. Even more, these variations could affect the trophic web, and alter the floodplain main subterranean
transfers of material and energy into the aquatic systems. channel transportation
lentic - lotic
environments
l. Introduction
Figure l. Main activities ofthe man that provoke stress in the aquatic systems.
1.1. Stressors in a Modern World
The aquatic continental ecosystems are tremendously susceptible and di\
In the freshwater ecosystem, the biological communities are in equilibrium and thus might be considered to be non-resilient to externa! forced cha
among all their biotic and abiotic components. However, the balances can vary whether from humans or climate. Rivers with floodplain and shallow lake
when the relative concentrations of the members are modified or any new intensely stressed during the Niña-Niño phenomenon. This provokes
elements are incorporated. These changes have origins in natural events or human oscillation of water to extreme levels, from desiccation to flood. In other w
activities and could be biological, physical or chemical. Their action can modify the lotic and lentic environmental stage is modified, the incorporating o
the interna! biochemical or cell frequency in sorne organs, thus provoking disease, dynamics of rivers or lakes to the abiotic and biotic elements in each p
an alteration of behavior, risks of failed reproduction, changes in the growth rate Moreover anthropogenic activities associated with changes in various ecosy~
or death. could induce the occurrence of diseases, mortalities, extinctions, habitat invas
Natural extreme events and the quantitative and qualitative increase of man- and species replacements; these function as sentinels and indicate that portio
made products that are used and discharged into environment cause constant risks the aquatic systems are under considerable stress.
to nature on all organizational levels (molecular, tissue, individual, population, The effects could be observed through assays of laboratory or m
and/or community). The broad categories of human activities are not limited to environments, using mesocosms or macrocosms, or by effects in the a
mining, dredging, fill, impoundment, point and non-point discharge, water communities. The measurements could be direct in individual organisrr
diversions, thermal additions, and other actions that contribute to source pollution. indirect through to their activities. Changes in the survival, metabo
Moreover, the introduction of potentially hazardous materials, exotic species, and composition or frequencies of cells in organs, which affect growth
the conversion of aquatic habitats in simple systems may eliminate, diminish, or reproduction, are sometimes observed. Furthermore, these variations coul
disrupt the functions and tolerance capacity ofthe biological components. affecting the trophic web, and their alterations could modify the materia
Due to the proximity of different human activities (industrial, farmland, and energy transference into the aquatic systems. At this point, you can antic
city), aquatic environments are affected more frequently by xenobiotic products. sorne problems facing the environmental toxicologist. On the one
These elements, when they reach the lotic and lentic environments through of the considerations are made in a comprehensive approach with the recognitio
direct spray, rain runoff or ground water connection, interact with each member of the effects upon high levels of biological organization (population, comm
the communities (Figure 1) [ 1] (2](3]. have more ecologically significance. The toxicologist recognizes that the e
and measurcmcnts at more simple levels (sub-organisms) are more scnsitiv
specific, b11t of' less ecological significance.
44 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

environment, provokes constant risk to the fauna and thus creates the necessity of
constant update studies._The continental aquatic environments, by its relative
instability, and proximity to different human activities (industrial, farmland, and
city) are more frequently affected by climatic phenomenon and xenobiotic cities
....
~>e:-;¡

aa&-
~~ rain
• · • • .·._ irrigation

products. When the actions reach the lotic and lentic environments interacts with
each member of the communities. The aim of this chapter is analyze and show fannland ·
the effects that it be observed in freshwater decapods due to natural and anthropic
stressors. The identification of the process that occurs in the environment is very
important, indicating when the species are affected by natural or anthropic superficial and
stressors. Even more, these variations could affect the trophic web, and alter the floodplain main subterranean
transfers of material and energy into the aquatic systems. channel transportation
Jentic - lotic
environments
l. Introduction
Figure l. Main activities ofthe man that provoke stress in the aquatic systems.
1.1. Stressors in a Modern World
The aquatic continental ecosystems are tremendously susceptible and di\
In the freshwater ecosystem, the biological communities are in equilibrium and thus might be considered to be non-resilient to externa! forced cha
among all their biotic and abiotic components. However, the balances can vary whether from humans or climate. Rivers with floodplain and shallow lake
when the relative concentrations of the members are modified or any new intensely stressed during the Niña-Niño phenomenon. This provokes
elements are incorporated. These changes have origins in natural events or human oscillation of water to extreme levels, from desiccation to flood. In other w
activities and could be biological, physical or chemical. Their action can modify the lotic and lentic environmental stage is modified, the incorporating o
the interna! biochemical or cell frequency in sorne organs, thus provoking disease, dynamics of rivers or lakes to the abiotic and biotic elements in each p
an alteration of behavior, risks of failed reproduction, changes in the growth rate Moreover anthropogenic activities associated with changes in various ecosy~
or death. could induce the occurrence of diseases, mortalities, extinctions, habitat invas
Natural extreme events and the quantitative and qualitative increase of man- and species replacements; these function as sentinels and indicate that portio
made products that are used and discharged into environment cause constant risks the aquatic systems are under considerable stress.
to nature on all organizational Jevels (molecular, tissue, individual, population, The effects could be observed through assays of laboratory or m
and/or community). The broad categories of human activities are not limited to environments, using mesocosms or macrocosms, or by effects in the aq
mining, dredging, fill, impoundment, point and non-point discharge, water communities. The measurements could be direct in individual organisrr
diversions, thermal additions, and other actions that contribute to source pollution. indirect through to their activities. Changes in the survival, metabo
Moreover, the introduction of potentially hazardous materials, exotic species, and composition or frequencies of cells in organs, which affect growth
the conversion of aquatic habitats in simple systems may eliminate, diminish, or reproduction, are sometimes observed. Furthermore, these variations coul
disrupt the functions and tolerance capacity ofthe biological components. affecting the trophic web, and their alterations could modify the materia
Due to the proximity of different human activities (industrial, farmland, and energy transference into the aquatic systems. At this point, you can antic
city), aquatic environments are affected more frequently by xenobiotic products. some problems facing the environmental toxicologist. On the one
These elements, when they reach the lotic and lentic environments through of the considerations are made in a comprehensive approach with the recognitio
direct spray, rain runoff or ground water connection, interact with each member of the effects upon high levels of biological organization (population, comm
the communities (Figure 1) [1] [2][3]. have more ecologically significance. The toxicologist recognizes that the e
and mcasurcmcnts at more simple lcvcls (sub-organisms) are more sensitiv
spccific. hut of kss ccological significancc.
46 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

1.2. South America and their Ecological Hydro Systems The rivers of "Del Plata" irrigate the lands of southern Brazil, Bolivia, Urugu
Paraguay and Argentina, and all types of effluents (treated or no-treated) :
South America is characterized by aquatic systems with large rivers and flood deposited into these rivers.
plains. The second largest basin is "Del Plata", whose area corresponds to more Del Plata has three main rivers, Parana, Paraguay and Uruguay, and a lat
than 17% of the continent's surface, and in which approximately 39% of the number of smaller tributaries that are stilllarge rivers based on their volumes a
human population live and 60% of production activities (e.g. industries and extensions (e.g. Iguazu, Pilcomayo, Bermejo, Salado). These rivers are mait
farmland) occur. In this region, most areas are industrially developed urban feed by rainfall, except those that originate in the Andes Mountains [4]. Lat
settlements and intensive farms, as well as artificial channels and reservoirs for cities and farmland occur throughout the paths of these rivers where herbicid
generating energy. The "Del Plata" system is one ofthose watersheds, perhaps the insecticides and fungicides are applied and industries use heavy metals
most developed in South America in relation to industrial and agricultura! petroleum products.
activities and the density of people in urban developments (Figure 2).

1.3. Stress on the "Del Plata" System

The list of elements that could cause stress on the del Plata system
unknown, as in any other basin of the world where urban development, farmit
and industries are present. It is unrealistic to consider all possible elements ti
occur in a basin, although there are ways to identify and measure these, becat
this approach is too expensive and unfeasibly time-consuming. But indirectly e
can also recognize the effects of the main economic activities. Sorne of tht
include severa! sources such as thermal stress in shallow lakes, desiccation (e
more intensive droughts during the Niña periods), salinity changes associated w
Niña-Niño periods, pH changes caused by aquatic vegetation decomposition, n'
fauna introduced by the human activities with exotic elements, either intentiot
(aquaculture) [5] or accidental (baggage ships) [6].
Anthropogenic causes correspond to the normal activities in the house
cities, industrial processes and agriculture, which are primarily responsible for 1
emission into the biosphere of organic and other compounds, such as deterge1
and biocides, petroleum derivatives, heavy metals, and others.
Biocides are a problem that currently signifies a threat to all biologi'
communities. The movement of pesticides through the soil particles causes 1
contamination of groundwater, especially in cracked soils where the filtrati
increases. In the case of aerosols and particles of these elements, upon enteri
the atmosphere they can be transported long distances with the movement of
masses and finally enter into aquatic environments as a result of precipitation
as a result of gas exchange between the air with water. In addition, biocic
hgurc 2. South 1\mcrica aml "Del Plata" basin (('), showing urban dcvclopmcnt (/\) and present in cultivated fields are often washed or leached by rain toward to t
IISl' land with l~mn and industrial ut:ti,itics (B). (irc¡ scalc indicutcs diiTcrcnccs in thc ncarby rivcrs, strcams or lakes. Moreover, they are directly applied on the surf<
rondillllll (ill):'h vahiL'S hlac" a1HIIo11 'ahll'S. 11 hik) ofthc water toeontrol aquatic plants ami invcrtcbratcs (mainly mosquito larvac'
 4X P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ... 49

Furthermor~, land use for urban development and recreation activities

2. Decapod Biodiversity
rcprescnts an immediate risk to the aquatic environment. Producís eliminated by
rnan and urban wildlife are stored in impervious surfaces (streets, sidewalks, Due to their ecological importance, numerical abundance, and sensitivity to a
rooflops, courtyards). They are washed by rain and dragged with the storm water, variety of toxicants and pollutants, the decapod crustaceans have been recognized
rcsulting in a negative impact on the aquatic environments that receive the runoff. as sensitive indicators. However, their application and use in such programs is
Thc urban rainwater runoff constitutes a major source of pollution and provides
limited to few regions where the knowledge about taxonomy and their natural
nutricnts for aquatic environments. The pH, conductivity and concentrations of
history have been undertaken, in addition to the background know.ledge of their
calcium, potassium, magnesium sulfate, total phosphorus, nitrates, nitrites, biological characteristics that permit their use in ecotoxicological studies.
ammonia, lead, zinc, total suspended solids and biological oxygen demand are
sorne elements that could be modified in the aquatic environment due to runoff. Table l. Decapods that inhabits at "Del Plata" systems in the southern
The levels of application of biocides on farmland vary between the minimum section indicating with asterisk the species used in assays
and maximum recommended values [7] but in sorne sites and times could exceed
thc concentrations of suggested active ingredients. Be especially careful, due to Vulgar
Orden Suborden lnfraorden Fa mil y Especie
na me
sorne studies it has been verified that agro toxics degradation rates could be
Dendrobranchiata Sergestidae Sergestidae A ce tes paraguayensis shrimp
similar for both laboratory and field surveys [8). But other available data indicates
that the biocides half-lives in aquatic environments have different ranges Palaemonets argentinus * prawn
Macrobrachium ama=onicum prawn
according the characteristics of each aquatic system [9). The products of
fumigations are washed and flow towards the rivers and streams when there is Caridea Palaemonidae M borellii * prawn

abundant rainfall, reaching values that could be higher than the LC 50 determined M.jelskii prawn

in different works. For example, the chlorpyrifos and endosulfan values registered Pseudopalaemon boliVieri prawn

in natural environments (chlorpyrifos, 0.45 11g L- 1 in water and 225 .8 11g Kg- 1 for Astacidae Parastacidae Parastacus pilimanus crayfish

suspended particles; endosulfan, 3 18 11g Kg- l in suspended particles) were greater Crabs or
Aegla parana
pancora
than the LC 50 values obtained in the laboratory for example to decapods, which Anomura Aeglidae
Crab or
are the most affected organisms [ 10). These levels may vary according the season, A. uruguayana * pancora
rainfall, and farm culture methods (insecticide application, grain type, and integral Crab or
use of the land). These investigators suggested that in many cases the A. platensis
pancora
concentration of pesticides in runoff or floodwater exceeded the water-quality Dilocarcinus pagei * True crab
criteria for freshwater established by the USEPA [11)[12)[13) (0.041 and 0.056 D. septemdentatus True crab
~tg L- l for chlorpyrifos and endosulfan, respectively). There may be a biological Poppiana argentiniana True crab
gradient downstream from where the xenobiotics enter the rivers [14], but Sylviocarcinus australis True crab
decapod populations or others zoological groups may reappear downstream, S. pictus True crab
considering that the degradation rate is variable in the time without aquatic Trichodactylida Trichodacty/u borel/ianus * True crab
vcgetation [ 15). Both laboratory and field studies have reported the microbial Brachyura
e T kensleyi True crab
dcgradation of biocides in aquatic environments [ 16). Sorne substances that are T panop/us True crab
rclcased by industries and cities into fresh waters or are washed off of the land T fluviati/is True crab
from rains may be potentially toxic depending on their nature and amount. Heavy
"'c.o
"O "'"'
E
O) Valdivia camerani True crab
lllctals are potential toxic elements to decapods, too. ;>,
u
"'
u
O)
oO) Zi/chiopsis col/astinensis * True crab
o o: Z oronensis True crab
50 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

Sorne species of freshwater environments have certain advantages and A

biological characteristics that allow them to be used in assays. Among then,
decapods crustaceans have been able to conquest freshwater environments from the B
Cretaceous [17]. But, only a few families can be considered completely freshwater
&00
in South America. Prawns of the Palaemonidae family, true crabs of the
Trichodactylidae family, anomuran crab or pancoras belonging to the family _ si)l) ~~
! .tOO

Aeglidae and the crayfish of Parastacidae family are the members of the decapods ¡ ,..
~ lOO
that live in the continental aquatic environments of"Del Plata" basin (Table 1) [18].
All integrants have similar characteristics in relation to their biology. Almost
,..
000 ~~~~~~-T~~.-
all families in this region are endemics offreshwater systems, with only the prawn J , M A M J J A S OtfO

families occurring in brackish-marine systems in addition to freshwater

environments. The integrant that is dependent on the satine environments to
e
ká:es
complete its life cycle is included in the Sergestidae family, Acetes paraguayensis
[19], and Palaemonetes argentinus also lives in brackish water [20].
F!ooploln Parono RNe<

2.1. Environment Type and its Relationship with the Decapods Modified of [22].

The decapods reach high densities in natural environments of "Del Plata" Figure 3. Schematic representation of a section of "Del Plata" systems showing the F
systems and these fluctuate according to time of year or season. The density River with its floodplain (A); normal water-cycle leve! ofthe Paraná River (B); tran~
variation in the populations is related to the reproductive cycle and flood pulse of section of the Paraná River showing the main channel and floodplain with lake
the rivers. This is due to the process of dilution and concentration of the secondary channels during flooding (high leve!) and falling (low leve!) phases (C).
populations and the water area viable for their development (Figure 3) [ 18].
The coastline area of a river, including the few meters into the terrestrial All these areas are highly risky because they are the targets of '
environments, known as aquatic terrestrial transition zone (ATTZ) [21], is the strip applications to control aquatic vegetation or animal vectors that develop the
where the majority of species of decapods lives and develops, and populations have correspond to the area that first receives the water runoff after rains. The me
highest individual numbers. The aquatic vegetation together with this first aquatic that the land is washed the particulate material with all associated product;
zone plays an important role in the regulation oftemperature during the summer and man has used is transported. Moreover, the sediment, with its physical
winter periods. Moreover, the aquatic vegetation provides a place of refuge and a chemical characteristics (particle size, electric charge, and origin), is at
source of important trophic resources for the group [22]. Furthermore, crabs are accumulate and remain immobilized in this area where it is availablr
common in areas with fine sediment because in this area structurally stable caves biological communities [23].
can be built. These crabs can find refuge from potential predators and avoid periods
of drought, maintaining moisture for long periods inside. Other groups (e.g.
Aeglidae, Parastacidae, sorne members of the genera Trichodactylus) have the
3. The Action of Stress and Reaction of the Decapods
capacity to live below stones in streams where they seek refuge and food.
A stress occurs and the fauna reacts (e.g. high temperature, extreme
water or xenobiotics). Moreover when the biocide reaches the wate
organisms are in contact with them, and the decapods, as components e
aquatic communitics. react to thcm. Thc rcactions dcpcnd on thc biotic and a'
conditio11s a11d thL· t:haracteristics of each spccics. the phasc of thcir lile
52 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

(reproductive, molt and development), the element types, concentrations, times

and frequency ofexposure [24][25].

3.1. The Case of the Xenobiotic: Input in Decapods

The crustaceans have an impermeable chitin exoskeleton, and only in the joints
between the articles, where the chitin is thin, is there sorne exchange between the
interna! and externa! parts of the body. Furthermore, the gills are thin chitin
structures where gas exchange and the movements of certain ions occur [26].
In addition, through the digestive system, other elements, which by their
configuration or size cannot enter the gills by active or passive diffusion, could
enter into the decapods. Finally, these inputs occur through the hepatopancreas
(Figure 4).
The ability of substances to enter each of these places is related to the size of
molecules and their ability to enter by facilitated diffusion or through carriers.
Sorne of these items are stored in the hepatopancreas, while others could form
complexes in the structure ofthe exoskeleton. Also, the gonads, muscles and other
organs can store xenobiotics, however these happen in much smaller quantities.
Heavy metals are trapped in the sediment and their input in the body of
decapods can occur when the material of the sediment is captured with the
mouthparts together with food. This occurs due to the process of supplanting of
the undeveloped (or absent) moll gastric by the sand (the sand has the functions of
the moll gastric) [27]. In this direct introduction to the digestive system, the heavy
metals are absorbed in the hepatopancreas [28] (Figure 5).
Figure 5. Hepatopancreatic cells (TEM) of the freshwater prawn Macrobrachium bore
Hepalopaucreas (A) showing high density granules corresponding to heavy metals and minerals inclusions.

1;.
8-cell with digestive vacuole; r. R- cell wtih lipidic vacuole; f. F-cells with abund

Food- sediment
---~_ ._·__¿~'-. . . • ~lls(I)_
_ .-e- 1
,_r. _ru_s_cle(A)
4é .
rough endoplasmic reticulum. In any cell is observed damage cell , desquamatization,
rupture organelles.
) '\. '-
(I)

( 5mm ~ ¡
3.2. Internal Changes in Decapods: Molecular Interactions

\r '
Space between joints
(I)
Gonads (A)
When xenobiotics enter into the body of the decapods, they interact \\
interna! molecules in different ways. Once a toxic chemical enters an organi!
severa! biochemical and physiologic process occur, attempting to challenge
toxic stress causcd by the pollutant.
Figun: 4 . Principal body part of frcshwatcr dccapods that pcrmit input (1) and
<ll:l:llllllliation (1\) of" IO\iC SllhSI<IIll:l:S
54 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

A mode of action of these compounds is the inhibition of acetylcholinesterase prawns are ablated in both eyestalks. The hormonal imbalances of the inhib
(AchE) activity, which causes death by hindering nerve function and ultimately molt hormone and salt deposition hormone are the main reasons for mo11
the muscle response [29][30]. This enzymatic system is common to all organisms [40][41][42][43]. That is, severa! xenobiotics could interfere with molt e
and is the reason that biocide compounds are known as nonspecific in their causing changes in the ecdysteroid-receiver (ECR). This is caused by
toxicity. interference of the ecdysteroid signa!, alterations in the ecdysteroidogenesis :
Regarding sub-lethal effects, sorne important physiologic processes often or the disposal of ecdysteroids [44].
associated with individual fitness have been recognized as being affected, e.g., Moreover, certain organs that have synthesizer roles and excretory func
reproduction, development, behavior, and growth [31]. Chlorpyrifos inhibited also have detoxification functions, e.g. the hepatopancreas, green gland
AchE in stage-5 grass shrimp eggs [32]. Moreover, AchE inhibition occurred in exoskeleton. In the first, the epithelium is composed of four cell types
prawns that inhabit natural environment in which chlorpyrifos and endosulfan, undergo ultrastructural changes depending on the quality of the environment
among other biocides, are found [30]. These cells increase their size and frequency according to their role in the o:
Moreover, severa! xenobiotics could be inducing chemical steroidogenesis, storing or synthesizing substances. The roof portian of the hepatopancre;
altering the mechanisms of ecdisteroid production, storage and disposal [33]. damaged by concentrated biocides (e.g. pyrethroid). Changes in the structun:
Also, biocides (e.g. heptachlor, endosulfan, carbaryl) alter the cytochromes and frequency of the F-cells indicate its participation in the detoxificatio1
heat-shock protein 70 (HSP70), and the heavy metals (e.g. Cadmium) function xenobiotic substances [46].
with similar mechanisms to those produced during a heat stress [34][35][36][37].
Heavy metals are absorbed in the hepatopancreas, interfering in energy A B
production by altering the enzymes associated with sulthydryl groups (SH groups)
[38]. Among these, it has been observed that cadmium produces sub-lethal effects
associated with an inhibitory activity in the transport of ions. Thus, it can interfere
with the metabolism of calcium (a required element to barden the new
exoskeleton) during molting and the regulation of sodium and other ions during
the ionic hyper-regulation [39].

3.3. Cell Modifications After Environmental Stress

Different modifications in the cells and tissues occur when the xenobiotic
enters into the decapods as a response of the body. Alterations in the Y -organ and
seno gland may occur, affecting the production and storage of the inhibitory molt
hormone, or more integrally, the neurohormonal system located in the eyestalks.
Neurotoxins could affect the synthesis process in the nerve cells and especially the
transmembrane sodium influx, mainly in the neurohormonal system. When the
concentrations are lower than lethal doses, the ecdysis cycle could be altered by
biocides (e.g. chlorpyrifos and endosulfan), thus increasing mortality and
Figure 6. Schematic draw of Palaemonetes argentinus indicating the normal F-cell
affecting the reproductive events. Moreover, in assays it was observed that a with mitochondria, RER, ribosome and nucleus, and the rest showing the deteri
single cypennethrin application did not affect the P. argentinus population in 48 in crease of hcpatopancreas aftcr exposed to di fferent concentrations of cypermethrin (
h. but was lcthal after 50 days. The residual effect is due lo alterations in the D). Thc dcsquamatizntion. apoptosis ccll ami frcqucncy ccll changcs are cvidcnt in t
neumhornwnal system located in thc cycstalk. Similar ciTccts occur when thc stagc
56 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

The hepatopancreas is the site of multiple oxidative reactions, being an organ In the R-cell, an electrogenic exchange occurs between Na 2+ and H+, and
that produces free radicals. Structurally, the cytoskeleton is destroyed when number of R-cells depend on the physiological characteristics, nutritional state
protein phosphorylation is increased after toxic exposure. Also, dissociation and the animals, and other seasonal features of the environmental (temperan
hepatocyst death could occur. The crustaceans are considered to have a system conductivity), suggesting that these are involved in the absorption of calci
where the immune reactions of the cells for defense are often followed by necessary for the exoskeleton. According the season, the R-cells are m
melanization processes, indicating the occurrence of the ROS scavenger. B-cells abundant, and their numbers correlate with the need for more calcium [54][55].
are involved in intracellular digestion and F -ce lis produce digestive enzymes. In Certain diseases can affect the hardness of the exoskeleton, especially dur
the case of microcystin presence, the F-cells are not affected by the exposure to the summer when the hepatopancreas plays an important role in the availabilit)
these toxins but the antioxidant defense system is activated after their exposure organic and inorganic nutrients [56]. When the hardness decreases, sorne para:
[4 7]. Moreover the exposure causes epithelial desquamation, necrosis and folded can cause malformations (Figure 7).
basal lamina. The last feature was also observed in individuals under nutritional Sorne elements such as cadmium can enter through the epithelium of the g
stress [48][49]. The basal lamina infoldings may be the result of the epithelial and digestive system. This is carried through membranes and through C
retraction and atrophy due to the loss of water at high salinities. Degenerative channels, Na2+¡Ca2+ exc hangers or th e Ca2+ ATPase. Moreover, t he mee hant5 o

desquamation ofthe tubular epithelium in the P. argentinus hepatopancreas was a of osmoregulation might interfere with the entry and accumulation of cadmiurr
pathological feature. According to Vogt [50], the process of desquamation in the the exoskeleton [57]. Polycyclic aromatic hydrocarbons (PAHs) are found
hepatopancreas of Penaeus monodon starts with cells lysis, in particular of R- effluents containing petroleum products. These cause changes in the structure
cells, and the neighboring cells protrude in small basolateral extensions, pushing lame llar gills and a decrease in the size of the hepatopancreatic ce lis [58]. On
the damaged cells into the tubular lumen and generating ulcerations. In P. other hand, the number of absorptive and fibrillar cells are modified significar
argentinus, the degenerative desquamation is a normal periodic event during the during the molt cycle, and these variations are associated with the A TPase :
molting cycle [51][52]. The increased desquamation produced by different Na 2+/Ka+ A TPase [55]. Moreover, Cathepsin L has a role in the food digest
stressors results in a high rate of cellloss that does not allow for the restoration of and has been found in the digestive vacuo le of the B-cells of the hepatopancr
the damaged tissue (Figure 6). [59]. In this relationship with the hepatopancreatic cells, pyrethroids can ind
It has been established that ce lis in the hepatopancreas of the crab can express the production of reactive oxygen species (ROS) and oxidative tissue. "
p-glycoprotein (P-gp) in transitional F- or B-cells, suggesting that these ce lis are interaction of ROS and cell membranes is the result of lipid peroxidation in
specialized for the accumulation and elimination oftoxic compounds [46][53]. membrane. The cells possess various mechanisms to protect themselves fr
oxidative damage, and among them glutathione plays an important n
Glutathione S-transferase (GST) is an important enzyme that catalyzes xenobi<
conjugations with the help of glutathione, facilitating their exit from the body.'
reduction in the levels of glutathione directly reflects the increased activity
GST. In tum, the leve! of GST in the hepatopancreas indicates the existing le
of oxidative stress. The hepatopancreas is a place where most detoxificat
processes occur. Sorne of the mechanisms of toxicity are the lipidic peroxidat
mediated by free radicals, inhibition of glutathione, induction of GST, inhibi1
of lactate dehydrogenase (LDH) and neurotransmission. Severa! intrinsic fact1
such as the speciticity of each species, the ability to tolerate the stress
pesticides, the size and sex, and extrinsic factors (e.g. pH, temperature, salir
and purity of biocide) are important for predicting the toxicity of pesticides [(
h gun: 7 . Frcshwater crab 7.ilchiopsis collastinensis x-ray photograph showing thc channel Among othcrs, thc polychlorinated biphenyls (PCB) action occurs in
produl·l·d h¡ parasitic crodcd in the abdomen ami thc rore (ustcrisks). hcpatopancrcas ami gonads, whcrc thcy wcrc rcgistcrcd in highcr conccntrati
SS P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

compared to the muscle [61]. In addition, some nutritional problems are related to neurotransmitters not only in the "pests" that they are intended to remove, but
vitamin E deficiency, which causes cell retraction, nuclear desquamation, and on the entire aquatic fauna. In experiments with crabs (T. borel/ianus), it
hypertrophy in the hepatopancreas [62]. observed that in the ftrst hour of exposure to these pollutants, little 01
movement occurs under sub-lethal and lethal concentrations of a pyreth
(cypermethrin). Then, involuntary motor activity begins. This activity
3.4. Response of Freshwater Decapods after Environmental Stress characterized by very rapid lateral movements and in some cases side jw
After several minutes, a time that varies in each animal, the crabs were lying
As toxins enter and interact with the molecules in the organism, the behavior, upwards pereiopods. In this position, the appendages shrink and str
growth, reproduction and survival ofprawns and crabs could be affected [46][63]. rhythmically. These actions occur with reduced frequency until death occurs.
Biological characteristics of this group permit its use in the ecotoxicological period covered from the beginning of the fast movements unti! death is 1:30 h
evaluation of different elements that could enter the aquatic environment, being a approximately, a time that varíes according to the characteristics of
good indicator of environmental stress. Some factors that affect the entry of any individual (nutrition, molt cycle, reproductive moment, development pt
stressors are the growth rate, respiration rate, excretion rate, weight, wet/dry rate,
(Williner & Collins, unpublished).
lipid contents, chemical assimilation efficiency and food assimilation efficiency. Evaluations have sorne subjectivity because they represent the interpret<
of the observations. For this reason it is necessary that a protocol is developt
3.4.1. The Behavior Level register each action according to direct observations or films. The time accor
to the species could vary between 1' and 5' minutes, indicating the typ
The first response to any change in the physical-chemical conditions is the
displacement (still, side, tilted, etc.).
escape behavior. If the individuals cannot do this in the time and space, then these
An indirect response related to the elements involved in the mobilizatic
animals are affected. This initial perception is made by chemosensory pegs on the
the autotomization of the appendages. In severa! experiences, the crabs ir
antennules (aesthetascs) [64].
inverted position caused by the toxic effects provoked the autotomization o
A group of crabs that belong to "Del Plata" System have the ability to walk
pereiopods or chelas.
on the Jand, with this being a possible way to escape to unaffected environments.
Previous works have reported abnormal behaviors, such as fast movem
There have been massive movements, such as when isolated ponds dry up due to
frequent jumping, erratic swimming, spiraling, convulsions, a tendency to es
very extreme temperature, conductivity, food and predator conditions [65]. While
from aquaria, and the secretion of mucus over the gill chamber [67] produce
some species (prawns) could escape by swimming or walking through water or
the exposure to endosulfan. Similar neurotoxic effects were observed i1
the sediment, other groups such as aeglas, lobsters and crabs forms caves and hide
argentinus when this species was exposed to endosulfan and chlorpy
under rocks to avoid natural changes that cause stress. These behaviors could be a
solutions, and these alterations permitted an assessment ofthe impact and Jeth
greater risk because many pollutants are trapped in the sediment and maintain a
sub-lethal effects of contaminants. Furthermore, the no-obser
closer interaction with these organisms [38]. Perhaps this will result more in a trap
effect concentration and lowest-observed-effect concentration (NOEC and L
that in a leak. However, the crabs can close their branchial camera for a large
values are used to determine sensitivity to the toxic action ofthese insecticide
time, and because their exocuticle is thicker, this avoids a rapid input into the
animal. The sensibility in these animals is lower than in prawns by their behavior.
If individuals have been unable to escape the stress, changes can be observed in
3.4.2. Osmoregulation and Metabolic Level
the form or intensity of locomotion. This change of behavior occurs when
Deviations in the decapad homeostasis associated with lethal and sub-!
decapods are already affected by something that interacts with neurotransmitters,
exposure reflect physiological alterations. Such modifications suggest an a
affecting the statocysts or mechanoreceptors in the setae of pereiopods [66]. For
mode of the toxicant and indicate the capacity to maintain homeos
example, in urban and agricultura! systems synthetic pyrethroids are used to
Physiological changes include impaired performance, such as swimming s
control insects. These compounds are characterized by the effects on
respiration, and excretion [24]. Besides, this stress is often reflected in imp
60 P. Collins, V. Williner and M. Montagna Contamination? Natural and Anthropic Stressors ...

cytologicat changes in the hepatopancreas as a main site of metabolism. The and the behavior of target and nontarget species, causing incremental chang
deterioration of the tubular epithelium could be evidenced by the loss of contact metabolic rate. These responses are attributed to neurotoxic effects :
between cells and the basal lamina [68], which has progressive effects of acetylcholinesterase inhibitions (AchE). In studies with the decapods
reduction in the metabolic function. The crustacean hepatopancreas is the most borellianus and P. argentinus), insecticides such as endosulfan, chlorpyrifos
important organ in the general economy because it serves as the main energy cypermethrin provoked the highest oxygen consumption values associated ·
reserve for growth and molting [68][69]. their constant movements (walking, erratic swimming, and back jumping).
Sorne freshwater decapods in the "Del Plata" system have the capacity to highest metabolic rates occurred during the first hours of the xenob
maintain homeostasis in seawater, such as P. argentinus in brackish water or exposures, when the oxygen consumption is probably a direct function of
extreme freshwater environments, those environments with a very low content in pesticide presence and the locomotory activity. The results have shown
salt (20 uScm). P. argentinus hyper-regulates at low salinities and hyper- intluence on physiological performance that impact the muscle activity and aJ
osmoconforms or isoregulates at higher salinities up to 32%o [70]. Under natural the respiratory capacities and energy expenditure (Figure 8).
conditions, the biological cycle of P. argentinus can be accomplished in either
170 ~--------------------------,
freshwater or brackish water [20]. T
- 160
In relation to the oxygen consumption, freshwater decapods demonstrate a
wide range of consumption levels when the animals are exposed to severa! types .,.~ 150

and concentrations of biocides. The highest metabolic rates can occur during the o"' 140
"'
§. 13 o
first hours of the exposure to biocide concentrations, when the difference in the e

oxygen uptake will probably have a direct relationship between the biocide i 120
E
presence and the locomotory activity. Those biocides with neurotoxic effects ~ 11 o
8 10 o .T
provoke first a constant activity, such as swimming and walking, similar to that ~ lK -.__ T
g: 90
mentioned in other animals by Salibian [71].
8o
1 ---;K
The reduction of dissolved oxygen (DO) is the result of oxygen consumption
(; 1
¡ - -<>uu ~g L" ·¡ 1
7o 1
by animals and the diffusion of oxygen from the air to the water [72]. The oxygen 1 1

o 07. o 12 o13 o 18 0.19 - 2~ 00

uptake of prawns (e.g. P. argentinus) with higher oxygen levels in water suggests Weight(g)
that there is a relationship between its consumption and the normal functioning of
the animal. Similar behavior was documented for crabs (e.g. T borellianus). On Figure 8. The relationship between oxygen consumption and weight of Trichodac
the other hand, the oxygen uptake by P. argentinus was similar that observed in T borellianus crab exposed to endosulfan concentrations (Zebra Ciagro ®): E 1: 625 Jlg
1 1
borellianus exposed toan insecticide [63]. E2 : 1250 Jlg L" ; E3 : 2500 11g L- [25]. The graph shows the intluence ofpesticide tox
Animal oxygen consumption is an appropriate measure to describe the on oxygen consumed in relation with the size ofthe animals.
respiratory capacity and estímate the metabolic rate of the organisms. Toxic
Naturally, there is a slowing down in the metabolism of animals accordin
compounds could affect this physiological parameter, indicating sorne
size, but the biocides cause an alteration of their metabolic patterns, affectin
abnormality or adaptive response in at least one of the biochemical pathways or
cellular and biochemical processes. The quantity of ammonia excreted
physiological processes governing the metabolism of the organisms. However,
crustaceans varíes according to externa! and interna! factors , including
any change in oxygen consumption depends on the nature, magnitude, and
injury, molting, temperature, and the presence of other animals [75], but may
persistence of toxic effects. Therefore, the effect of compounds on metabolic rate
include the factors of swimming activity as a retlection of the biocide action
provides an index for stress to infer a mode of action of the chemicals on the
metabolism alteration.
homeostasis or normal functioning. Moreover, alterations in the oxygen
consumption could be used for biomonitoring the potentially toxic effects of
chemicals [73][74]. For example, severa! pesticides affect respiration, swimming