TROPICAL ENTOMOLOGY

"OF RICE AND MEN": STARTING A

DISCUSSION ON CARBOFURAN
UTILISATION IN BRAZIL

2020-2021 // ELISSA PELSMAEKERS

1
TABLE OF CONTENTS

I. INTRODUCTION 3

II. CASUAL, CUSTOMARY CARBOFURAN 4

CARBO-WHAT?

TAKE IT WITH A GRAIN OF RICE: A CASE STUDY

UNFORTUNATE ENVIRONMENT

NATURE AND EFFECTS

ACTUALLY WE ARE NOT SURE...

III. BOB DOES NOT USE PESTICIDES, BE LIKE BOB 9

BIOPESTICIDES

BIOCONTROL

OH-MICROBIAL DEGRADATION

IV. WHAT NOW? 13

2
Over the past few decades, the human population has grown immensely and has led to an
increase in food production demands (POPP et al., 2012). Pesticides are progressively used
more due to intensification and expansion of agriculture in the tropics (CHELINHO et al., 2012;
BUCH et al., 2012; BASSANI et al., 2018). In general, pesticides have been around for a long
time. But it was not until 20 years after introduction of synthetic pesticides in the 1940s,
questions regarding their use rapidly started to emerge (VAN DER WERF et al., 1996).

Since 2008, Brazil has become one of the largest consumers to ensure the quality of
production (CHELINHO et al., 2012; BUCH et al., 2012) (BASSANI et al., 2018). In 2015, a
whopping 899 million litres of pesticides were used on the cultivated crops (PIGNATI et al.,
2017). While the European Union banned a great number of pesticides found dangerous to
the environment and human health, Brazil is still the front runner in pesticide use with its
market estimated at 10 billion US dollars a year (BASSANI et al., 2018). One of these (still
permitted) pesticides is carbofuran, a carbamate which serves as a control method against a
wide variety of pests (GUPTA, 1995; MISHRA et al., 2020). Even though many concerns have
already been raised about its extreme toxicity and environmental impacts, it is still massively
used in many countries (GUPTA, 1995; MISHRA et al., 2020; PESSOA et al., 2011).

This essay aims to provide contextual information on pesticides and pesticide pollution, with
particular mentioning of carbofuran use in Brazil. Its toxicological effects in organisms are
going to be addressed and alternative options to reduce pesticide use will be discussed, and
critically analysed.

3
Carbofuran is a commonly used pesticide, which gained much interest, especially in the
tropics, due to its broad-spectrum functionality (GUPTA, 1995). However, together with the
gain in enthusiasm came also increased curiosity concerning carbofuran toxicity (GUPTA,
1995; MISHRA et al., 2020). The utilisation of carbofuran has already been prohibited in
several countries. However, currently it is still being used in agricultural activities in certain
parts of the tropics (MISHRA et al., 2020). In Brazil, carbofuran has not been banned yet and is
widely used in rice and sugar cane plantations (GUPTA et al., 1989). But what is carbofuran
exactly, and what are the environmental and health impacts related to its use?

First introduced by the FMC corporation in 1967, carbofuran (2,3-dihydro-2,2-dimethyl-7-

benzofuranol methylcarbamate) is an anticholinesterase carbamate and serves as a pest
control method against insects and nematodes. It belongs to class I of N-methyl carbamate
pesticides with ester and amide linkage and is derived from carbamic acid (Fig. 1) (GUPTA,
1995; MISHRA et al., 2020). It is mostly applied directly in the soil or water, through which it
can be absorbed by the plant and translocated to other structural parts. Plant-sucking pests
are affected due to the chemical being present in the plant’s phloem sap (MISHRA et al.,
2020).

4
 Figure 1: Chemical structure of carbofuran (Sigma Aldrich)

In the past few decades, carbofuran has known a tremendous increase in production and
utilisation due to its broad range of applications against insects (MISHRA et al., 2020).
Carbamates are more widely used chemicals in comparison to other products because of the
extreme toxicity of organophosphates (OPs) and persistence of organochlorines (GUPTA,
1995). However, carbofuran also seems to display high toxicity to organisms and persistence
in the environment, and due to the increase in utilisation, contamination of air, food, soils
and water is becoming a matter of concern (GUPTA, 1995; MISHRA et al., 2020).

Even though carbofuran was banned by the European Union and the United States due to its
toxicity and environmental effects, presently, it is still heavily used in several countries,
including Brazil (MISHRA et al., 2020; PESSOA et al., 2011).

Around 60% of the rice grown in Brazil comes from the southern region (Rio Grande do Sul
and Santa Catarina) where pre-germination and irrigation by flooding techniques are used
for cultivation. The rice fields are subjected to intensive farming activities which makes the
areas prone to the development of pests, weeds and diseases (VIEIRA et al., 2016; BAUMART et
al., 2011). Additionally, flooding of the fields creates an ideal environment for pests,
including several coleopterans and dipterans, to thrive (BAUMART et al., 2011). Consequently,
pesticides are eagerly used to ensure the profitability of the crops (VIEIRA et al., 2016;
BAUMART et al., 2011).

5
Oryzophagus oryzae (Coleoptera: Curculionidae) or the rice-infesting weevil is one of the
major rice crop pests in Brazil, introduced around the 1930s (Fig. 2) (PLESE et al., 2005;
MARTINS et al. 2009; OLIVEIRA et al., 2012). It causes a loss of approximately 80 million US
dollars per year by lowering the yields by around 10-18% (OLIVEIRA et al., 2012).
After flooding, the larvae damage the roots of rice, which causes the plants to take up fewer
nutrients. Carbofuran is widely used to control this pest. It is applied in granular form by
adding it to the side of the plant stems or directly in the irrigation water (PLESE et al., 2005).

Figure 2: Image of the rice-infesting weevil (Oryzophagus oryzae). A: adult; B: larvae; C: damaged root system of rice
plant (MARTINS et al., 2009).

More intensive use of pesticides comes at a cost. Studies conducted on how much of the
pesticide applied ended up reaching the targeted pests displayed only 0,3% of the pesticides
coming in contact with their targets (VAN DER WERF et al., 1996). Most of the pesticides are
sprayed on top of the leaves or soil by planes or other machines, injected into the ground or
are sprinkled as granules. These application methods account for a great loss of pesticides in
the environment due to nonspecific treatment (VAN DER WERF et al., 1996).
There are several ways of pesticides ending up in the environment. The volatilisation of
pesticides in the atmosphere occurs when they are directly applied to the surface of the
plant. This can lead to uptake by the plant through the leaves or the soil, which exposes
organisms to pesticides after consuming it. Additionally, raining or flooding of agricultural

6
grounds may cause surface runoff or leaching and pollution of ground and surface water
(VAN DER WERF et al., 1996).

Over the years, it has been found that carbofuran is extremely lethal to organisms, including
mammals, birds and fish (GUPTA, 1995; MISHRA et al., 2020). Carbofuran acts as an inhibitor of
acetylcholinesterase (AChEase) and butyrylcholinesterase in mammals. It increases
acetylcholine at the nerve synapses leading to cholinergic toxicity and toxic symptoms. It is
based on reversible competitive inhibition of AChEase in which the serine residue in the
enzyme’s active site is carbamylated (GUPTA, 1995; MISHRA et al., 2020).

In humans, it disrupts endocrine activity and causes neurotoxic, cytotoxic, genotoxic and skin
problems. Additionally, it is associated with reproductive disorders, and it is identified as
mutagenic (MISHRA et al., 2020). It can enter the body through inhalation, ingestion (high
toxicity) and absorption through the skin (low toxicity). Oxidative stress generated by the
chemical can eventually lead to lipid peroxidation, inhibition of enzymes present in red
blood cells and affect Na+ K+ ATPase activity in the membrane of cells. General symptoms of
carbofuran intoxication include weakness, nausea, vomiting, headache and diarrhoea
(GUPTA, 1995; MISHRA et al., 2020).

In aquatic organisms, disruption of immune cells, biochemical pathways and enzymatic

activities are probable. Exposure can also cause behavioural abnormalities, including loss of
balance, feeding difficulties, decreased growth rate and affected swimming performance
(MISHRA et al., 2020). In a study conducted by Vieira et al. (2016) on risk assessment of
pesticides used in Brazilian rice fields, it was revealed that microcrustaceans were one of the
more severely influenced organisms in freshwater bodies.

Research conducted by Mansano et al. (2016) on toxical effects of carbofuran in neotropical

Ceriodaphnia silvestrii, which displays a wide distribution in South America, showed a
decrease in survival rate and detrimental effects on their reproductive system. Another

7
study done by Pessoa et al. (2010) exposed Nile tilapia Oreochromis niloticus to carbofuran
for 96 hours. There was a large decrease visible in the activity of cholinesterase. Additionally,
the fish showed a decrease in swimming speed, feeding behaviour, growth and clarity of
vision.

Additionally, the simultaneous use of certain chemicals can enhance the toxic effects
generated in organisms after exposure (synergism) (GUPTA, 1995; MANSANO et al., 2020). In
Brazil, combinations of several pesticides have been found in freshwater ecosystems
(MANSANO et al., 2020). In the case of carbofuran, not much information is available on
additive and synergistic effects with other pesticides (GUPTA, 1995). However, studies
conducted by Iyaniwura (1990 & 1991) showed additive effects on AChe activity with two
other types of carbamates (oxamyl and propoxur). Additionally, rats treated with a
combination of carbofuran and an organophosphate compound (iso-OMPA), showed more
severe hypercholinergic signs than during the separate treatments (GUPTA et al., 1989).
Assessments of the risks involved should therefore also include interactions of carbofuran
with other chemical compounds (MANSANO et al., 2020).

Furthermore, carbofuran can also affect regular soil properties, including fertility,
respiration, nitrogen and phosphate mineralisation, plant enzymatic activities and microbial
biomass. A decrease in microbial growth could, in turn, affect the activity of enzymes
needed for nutrient cycling, such as alkaline phosphatase (MISHRA et al., 2020).
Consequently, organisms living in the soil are affected by these changes. According to
research, collembolans, earthworms and enchytraeids are the most impacted groups
(CHELINHO et al., 2012). Significant changes in microarthropod communities of Acari and
Collembola extracted from Portugal and Brazil, important in nutrient cycling and
decomposition, were observed by Chelinho et al. (2014). The groups were exposed to
carbofuran under laboratory conditions. The total number of individuals present in the
collembolan communities decreased significantly after carbofuran application, with
communities extracted from Brazil showing more sensitivity towards the pesticide. Mites
showed an increase in abundance, probably due to a decrease in their predators.

8
There is still little known about the impacts of carbamate pesticides in the tropics compared
to temperate ecosystems. Climatic conditions differ tremendously, which can lead to
misconceptions about the proper use and effects of pesticides (CHELINHO et al., 2012). There
may be a higher risk of environmental contamination and enhanced toxicity of these
chemicals in tropical surroundings due to different climatic factors. Furthermore, excessive
rainfall could increase surface runoff and leaching of contaminants in soils, especially due to
the high solubility of carbofuran in water (CHELINHO et al., 2012; VIEIRA et al., 2016; ROCHA et
al., 2018). However, the extent of contamination of groundwater and water bodies
surrounding the treated agricultural fields is not yet quantified (ZANELLA et al., 2011).

Additionally, the use of pesticides in the tropics is scarcely managed or restricted, and their
application does not always follow the imposed rules and regulations due to the
underdevelopment of national legislation (CHELINHO et al., 2012; VIEIRA et al., 2016). Because
of the importance of agriculture to the economy of Brazil, laws seem to be implemented to
ensure an even bigger production (e.g. the so called “Poison Law” project from 2002; BASSANI
et al., 2018). The high amounts of carbofuran residues detected in the Brazilian environment
illustrate that agricultural practices are still not pursuing the advised crop management
approaches, e.g. minimum water retention after pesticide application (VIEIRA et al., 2016).
Furthermore, pesticides are still administered by plane or tractors, which are more
innaccurate forms of application and creates even more risks of exposure for workers
(PIGNATI et al., 2017).

Because of the high toxicity of many pesticides, alternatives are being developed to reduce
their use (GLARE et al., 2012; SAMPAIO et al., 2010; HAJEK, 2004). In this chapter, the
application of biopesticides and natural enemies against pests will be critically analysed, and

9
questions about their practicality, benefits and drawbacks will be raised. Could there be
alternatives for the use of carbofuran and could methods of carbofuran clean-up be
developed?

Biopesticides are products that are based on living organisms, such as microbes and their
metabolites (bacteria, viruses, fungi, protozoa), botanical compounds (e.g. plant extracts)
and semiochemicals (e.g. pheromones) (GLARE et al., 2012). Food production and other
related activities have strongly relied on synthetic pesticides for a long time. However,
alternative pest management procedures are slowly making their way into the agricultural
world (CHANDLER et al., 2011). Reasons for the emergence of these biopesticides include rapid
emergence of pest resistance and environmental and human-related risks (GLARE et al.,
2012).

However, their use is still limited due to several drawbacks, and they only make up a small
number of pest control products sold. There is a need for improvements of formulas, they
are more expensive and generally act slower than the traditional pesticides, registration
procedures are challenging, and many biopesticides have a highly specific activity (GLARE et
al., 2012; VILLAVERDE et al., 2014). This means only a single pest is targeted, which limits their
use. Additionally, there is a large difference in utilisation between biopesticides and
synthetic pesticides. Their handling and application are still unknown to the majority of
agriculturists (GLARE et al., 2012).

With biological control, regulation of organisms by their natural enemies can be achieved. It
represents an alternative method of pest control, said to have lower impacts on the
environment and human health than the use of pesticides (SAMPAIO et al., 2010; HAJEK, 2004).

10
This is based on regulation processes within food webs, already occurring in the natural
environment (HAJEK, 2004).
Insects as natural enemies are known as the entomophagous group, which include predators
and parasitoids. Microorganisms represent the entomopathogens, including bacteria, fungi
and viruses (SAMPAIO et al., 2010).

Although this option seems to be more beneficial than using chemicals, it leads to some
controversy among and conflict between biological control practitioners and
conservationists (HAJEK, 2004).
The first type of biological control occurred through the use of ladybugs (Coleoptera;
Coccinellidae). However, the release of an exotic species of ladybugs may affect native
species, and due to non-specificity, it may reduce other non-targeted beneficial organisms
(HAJEK, 2004). Additionally, control agents helpful against certain pests may be pests
themselves in diverse contexts. For example, the introduction of the Halloween lady beetle
Harmonia axyridis in the USA was used to control aphids. They found their way into and
aggregated in populated areas. Inhabitants eventually started looking at the beetles as pest
themselves (HAJEK, 2004).

Bioremediation is a way of treating contaminated material (e.g. soils and water) and
removing toxic compounds by use of microbial degradation. Bacteria, algae and fungi can
degrade certain pesticides in the environment, including carbofuran (GUPTA, 1995; MISHRA et
al., 2020). Due to the excessive use of carbofuran in agricultural fields, resistance in
microorganisms has occurred. Consequently, they found ways to deal with pesticide-related
stress and develop mineralisation and degradation techniques to detoxify carbofuran
(MISHRA et al., 2020).

Isolated carbofuran metabolising bacteria include Archromobacter sp., Rhodococcus sp.,

Enterobacter sp., Burkholderia sp., Bacillus sp. And Cupriavidus sp (MISHRA et al., 2020). One
of the processes of metabolism encompasses the conversion of carbofuran to 4-

11
hydroxycarbofuran by the inducible oxidative hydroxylase enzymes or hydrolysis at the
furanyl ring (Fig. 3) (GUPTA, 1995; MISHRA et al., 2020). For these bacteria, carbofuran is used
as a primary source of nitrogen and carbon.
Fungi are also important in pesticide degradation. They contain esterases, hydrolases,
laccases and peroxidases that mineralise pollutants (MISHRA et al., 2020).

However, research on microbial degradation is still in its infancy, and not much information
about degradation pathways and their functions are available. More research is needed to
understand the potential of these pesticide clean-up techniques (MISHRA et al., 2020).

Figure 3: Degradation of carbofuran by an oxidative metabolic pathway (MISHRa et al., 2020).

12
Although pesticides cause significant public health issues and severe environmental damage,
they are still widely used. Reducing our dependence on these chemicals should be a priority.
Could the implementation of alternatives work? Could bioremediation be our top solution?
However, even though biopesticides take part in the development of a more sustainable way
towards crop protection, there is still little information on the long-term effects (e.g. the
potential development of biopesticide resistance by insects). Pesticide risk assessment is
primarily based on short-term acute effects. More research needs to be conducted on long-
term effects and risks towards natural enemies. Furthermore, there is insufficient
information about most natural enemies used in biocontrol, which makes it challenging to
use these organisms beneficially. If there is so little known about the real environmental
impacts, why use it? More attention should be transferred from the problems created by the
pest towards the impacts of the introduction of biological control species on the
environment.
So, maybe it is time to start asking ourselves different kinds of questions. Why are new pests
emerging? Why is the distribution of certain pests shifting towards new regions? Why are
we looking for solutions to problems if we can stop this from happening in the first place?

The reason why I chose this subject is that as a scientist, it is necessary to start a discussion.
Why are we looking for solutions to already existing problems? To say it metaphorically, we
are putting our finger in a dyke. We should start looking for ways to change our behaviour
and sustainably reduce our dependence on pesticides before the issue emerges, instead of
trying to mitigate the damage already done. I know, “easier said than done”, “it is what
everybody is saying nowadays”. Well, good. We started the conversation, now let us do
something about it. That being said, mapping out new emerging pests is important as well. It
can actually help us in figuring out what needs to be changed and how we can tackle these
changes and finding strategies to stop them from happening.

13
BASSANI, D., L.R. DA SILVA, L. BIRK, S.C. DE OLIVEIRA. E. DELLAGRAVE & T.F. DE OLIVEIRA. (2018).
Pesticides in Brazil: A Viewpoint about the Poison Law. Journal of Agricultural and Food
Chemistry. doi: 10.1021/acs.jafc.8b05669.
BAUMART, J.S., M.M. DALOSTO & S. SANTOS. (2011). Effects of carbofuran and metsulfuron-
methyl on the benthic macroinvertebrate community in flooded ricefields. Acta Limnologica
Brasiliensia. 23(2): 138-144.

BUCH, A.C., G.G. BROWN, C.C. NIVA, K.D. SAUTTER & J.P. SOUSA. (2012). Toxicity of three pesticides
commonly used in Brazil to Pontoscolex corethrurus (Müller, 1857) and Eisenia andrei
(Bouché, 1972). Aplied Soil Ecology. 69: 32-38.

Carbofuran. Sigma Aldrich. Retrieved November 11, 2020, from

https://www.sigmaaldrich.com/catalog/product/aldrich/426008?lang=en&region=US

CHANDLER, D., A.S. BAILEY, M. TATCHELL, G. PRINCE, J. GREAVES & W. GRANT. (2011). The
development, regulation and use of biopesticides for integrated pest management.
Philosophical Transactions of The Royal B Biological Sciences. 366(1573): 1987-1998.

CHELINHO, S., I. LOPES, T. NATAL-DA-LUZ, X. DOMENE, M.E. TENORIO NUNES, E.L. GAETA ESPINDOLA, R.
RIBEIRO & J.P. SOUSA. (2012). Integrated ecological risk assessment of pesticides in tropical
ecosystems: A case study with carbofuran in Brazil. Environmental Toxicology and Chemistry.
31(2): 437-45.

CHELINHO, S., X. DOMENE, P. ANDRÉS, T. NATAL-DA-LUZ, C. NORTE, C. RUFINO, I. LOPES, A. CACHADA, E.

ESPINDOLA, R. RIBEIRO, A.C. DUARTE & J.P. SOUSA. (2014). Soil microarthropod community testing:
A new approach to increase the ecological relevance of effect data for pesticide risk
assessment. Applied Soil Ecology. 83: 200-209.

GLARE, T., J.R. CARADUS, W. GELERNTER, T.A. JACKSON, N.O. KEYHANI, J. KOHL, P.G. MARRONE, L. MORIN
& A. STEWARD. (2012). Have biopesticides come of age? Trends in Biotechnology. 30(5): 250-
258.

GUPTA, R.C. (1995). Carbofuran toxicity. Journal of Toxicology and Environmental Health.
43(4): 383-418.

GUPTA, R.C. & W. KADEL. (1989). Concerted role of carboxylesterases in the potentiation of
carbofuran toxicity by iso-OMPA pretreatment. Journal of Toxicology & Environmental
Health. 26(4): 447-457.

14
HAJEK, A.E. (2004). Natural enemies, an introduction to biological control. In: Natural
Enemies, An Introduction to Biological Control, Cambridge University Press, Cambridge.
https://doi.org/10.1017/CBO9780511811838

IYANIWURA, T.T. (1990). Mammalian toxicity and combined exposure to pesticides. Veterinary
and human toxicology. 32(1): 58-62.

IYANIWURA, T.T. (1991). Relative inhibition of rat plasma and erythrocyte cholinesterases by
pesticide combinations. Veterinary and human toxicology. 33: 166-168.

MANSANO, A.S., R.A. MOREIRA, H.C. DORNFELD, L.G. DINIZ, E.M. VIEIRA, M.A. DAAM, O. ROCHA & M.H.
SELEGHIM. (2016). Acute and chronic toxicity of diuron and carbofuran to the neotropical
cladoceran Ceriodaphnia silvestrii. Environmental Science and Pollution Research. 25(14):
13335-13346.

MANSANO, A.S., R.A. MOREIRA, H.C. DORNFELD, E.C. FREITAS, E.M. VIEIRA, M.A. DAAM, O. ROCHA &
M.H. SELEGHIM. (2020). Individual and mixture toxicity of carbofuran and diuron to the
protozoan Paramecium caudatum and the cladoceran Ceriodaphnia silvestrii.
Ecotoxicological Environmental Safety. doi: 10.1016/j.ecoenv.2020.110829

MARTINS, J.F. DA S., J.A. F. BARRIGOSSI, J.V. DE OLIVEIRA & U.S. DA CUNHA. (2009). Situação do
Manejo Integrado de Insetos-praga na Cultura do Arroz no Brasil. Embrapa Clima
Temperado. ISO: 1806-9193.

MISHRA, S., W. ZHANG, Z. LIN, S. PANG, Y. HUANG, P. BHATT & S. CHEN. (2020). Carbofuran toxicity
and its microbial degradation in contaminated environments. Chemosphere. 259: 127419.

OLIVEIRA, C.M., A.M. AUAD, S.M. MENDES & M.R. FRIZZAS. 2012. Economic impact of exotic insect
pests in Brazilian agriculture. Journal of Applied Entomology.
https://doi.org/10.1111/jen.12018

PESSOA, P.C., K.H. LUCHMANN, A.B. RIBEIRO, M.M. VERAS, J.R. CORREA, A.J. NOGUEIRA, A.C. BAINY &
P.S. CARVALHO. 2011. Cholinesterase inhibition and behavioral toxicity of carbofuran on
Oreochromis niloticus early life stages. Aquatic Toxicology. 105(3-4): 312-320.

PIGNATI, W.A., F.A. DE SOUZA E LIMA, S.S. DE LARA, M.L. CORREA, J.R. BARBOSA, L.H. DA COSTA LEAO &
M.G. PIGNATTI. (2017). Spatial distribution of pesticide use in Brazil: a strategy for Health
Surveillance. Ciencia & Saude Coletiva. 22(10): 3281-3293.

PLESE, L., L. PARAIBA, L.L. FOLONI & L. TREVIZAN. (2005). Kinetics of carbosulfan hydrolysis to
carbofuran and the subsequent degradation of this last compound in irrigated rice fields.
Chemosphere. 60(2): 149-56.

POPP, J., K. PETO & J. NAGY. (2012). Pesticide productivity and food security. A review.
Agronomy for Sustainable Development. 33: 243-255.

ROCHA, O., A.J. NETO, J.C. DOS SANTOS LIMA, E.C. FREITAS, M. MIGUEL, A. DA SILVA MANSANO, R.A.
MOREIRA & M.A. DAAM. (2018). Sensitivities of three tropical indigenous freshwater

15
invertebrates to single and mixture exposures of diuron and carbofuran and their
commercial formulations. Ecotoxicology. 27(7): 834-844.

SAMPAIO, M.V., V. BUENO, L.C.P. SILVEIRA & A.M. AUAD. (2010). Biological control of insects pests
in the tropics. In: K. Del Caro (ed), Encyclopedia of Life Support Systems, UNESCO/EOLSS,
Paris.

VIEIRA, D.C., J.A. NOLDIN, F.C. DESCHAMPS & C. RESGALLA JR. (2016). Ecological risk analysis of
pesticides used on irrigated rice crops in southern Brazil. Chemosphere. 162: 48-54.

VILLAVERDE, J.J., B. SEVILLA-MORAN, P. SANDIN-ESPANA, C. LOPEZ-GOTI & J.L. ALONSO-PRADOS. (2014).

Biopesticides in the framework of the European Pesticide Regulation (EC) No. 1107/2009.
Pest Management Science. 70(1): 2-5.

VAN DER WERF, H.M.G. (1996). Assessing the impact of pesticides on the environment.
Agriculture, Ecosystems & Environment. 60(1-2): 81-96.

ZANELLA, R., M.B. ADAIME, S.C. PEIXOTO, C.A. FRIGGI, O. PRESTES, M.S. LUIZ DE OLIVEIRA, E. MARCHESAN,
L.A. AVILA, E.G. PRIMEL. (2011). Herbicides Persistence in Rice Paddy Water in Southern Brazil.
Environmental Science. doi: 10.5772/32727.

16