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Polysaccharides as Eco-Nanomaterials for Agricultural Applications

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Polysaccharides as Eco-Nanomaterials for
Agricultural Applications

Danila Merino, Claudia Casalongué, and Vera A. Alvarez

Abstract
The great growth of the world population, climate change, and environmental
issues related to the accumulation of pesticides and fertilizers are the main
problems facing the agricultural sector. To provide solutions, new innovative
developments committed to caring for the environment are required. Nanotech-
nology has demonstrated great potential to provide new solutions to the many
challenges faced by other sectors including agriculture.
The aim of this work was to carry out an updated version of the problems
facing the current agricultural sector and show how polysaccharide based eco-
nanomaterials might represent a promising solution. Polysaccharide applications
for the controlled release of agrochemicals and promotion of plant growth are
discussed. Finally, legal aspects on the development of these types of composite
materials and their commercial insertion are studied.

Keywords
Nanoparticles • Polymers • Agrochemicals • Polysaccharides • Chitosan • Algi-
nate • Regulations

D. Merino • V.A. Alvarez (*)

Grupo de Materiales Compuestos Termoplásticos (CoMP), Instituto de Investigaciones en Ciencia y
Tecnología de Materiales (INTEMA), Universidad Nacional de Mar del Plata (UNMdP) y Consejo
Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina
e-mail: alvarezvera@gmail.com; alvarezvera@fi.mdp.edu.ar
C. Casalongué
Grupo de Fisiología del Estrés en Plantas, Instituto de Investigaciones Biológicas (IIB),
Universidad Nacional de Mar del Plata (UNMdP) y Consejo Nacional de Investigaciones
Científicas y Técnicas (CONICET), Buenos Aires, Argentina

# Springer International Publishing AG 2018 1

L.M.T. Martínez et al. (eds.), Handbook of Ecomaterials,
https://doi.org/10.1007/978-3-319-48281-1_124-1
2 D. Merino et al.

Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Polysaccharides as Controlled Release Systems for Agrochemicals . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Chitosan and Alginate-Based Nanomaterials for Promoting Plant Growth and Protection
Against Stresses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Legal Aspects and Current Regulation on Nanomaterials Usage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Nanotechnology Risk Assessment and Regulation in the EU and Worldwide . . . . . . . . . . . . . . 13
Regulatory Issues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Current Nanomaterials in the Agricultural Field (Commercial and in Developing) . . . . . . . . . . . . 14
Conclusions and Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Introduction

Agriculture includes the set of practices and knowledge that allow farming. The
beginning of it back to approximately 12,000 B.C. Since then, it has conducted to
major changes that led to the establishment of human beings and the development of
trade and urbanization. Agricultural activity is the nexus that directly connects the
environment with humans and the main cause of climate change and detriment of
ecosystems [1]. Over time, the increasing demand for food has encouraged the
excessive use of agrochemicals which has generated a detrimental effect on the
environment and health. The word “agrochemicals” is often used to refer to
chemicals such as pesticides, herbicides, and fertilizers, among others [2]. In this
way, agriculture in twenty-first century is facing major challenges to meet the
growing food demand of an increasing global population, avoiding negative impacts
on soil and human health. The Food and Agriculture Organization of the United
Nations includes, among the objectives of its 2030 agenda, achieving the imple-
mentation of sustainable agriculture practices that will eradicate hunger and improve
food security and nutrition, and at the same time maintaining ecosystems and
improve soil quality [3].
Nanotechnology is the science that contemplates the understanding and manip-
ulation of matter at the nanoscale, where new and unique physical properties appear
bringing with them wide spectrum of potential applications [6]. In this context,
scientific advances and the emergence of nanotechnology have allowed, in part, the
development of the so-called “precision agriculture,” which aims to minimize costs
and resources and to maximize yields with nanoscale control using nanosensors and
intelligent delivery systems [4, 5]. From its understanding, nanotechnology has been
widely exploited by man in the pharmaceutical, medical, electronics, cosmetic, and
catalysis fields, while agriculture, compared to other industrial sectors, has been
relatively marginalized [4, 7]. In the last decade, however, there was a noticeable
growth in the number of scientific publications and patents in agro-nanotechnology
[4]. The application of this powerful tool in plant science is known as phyto-
nanotechnology. It mainly contemplates systems that produce controlled release of
biomolecules with specific temporal and spatial patterns [7]. This topic will be
Polysaccharides as Eco-Nanomaterials for Agricultural Applications 3

discussed in more detail in the section “Polysaccharides as controlled release

systems of agrochemicals.”
Conventionally, the application of agrochemicals is done by spraying or soaking.
This technique consists of its direct application on plants and soil and entails great
environmental and economic disadvantages. A large percentage of the active sub-
stances applied are quickly lost by leaching, degradation (photolytic, hydrolytic,
and/or biological), and volatilization and only a small amount, often below the
minimum required to fulfill their function, manages to reach the site of interest.
For this reason, in order to maintain its biological efficiency and concentration at the
appropriate levels, periodic applications of agrochemicals in excessive concentra-
tions are usually carried out, which implies high costs, low efficiency, and serious
environmental and health problems [2, 5].
As mentioned above, nanotechnology has allowed the development of new
alternatives to traditional methods of applying agrochemicals. It is the use of
nanoscale or nanostructured materials that act as vehicles for the controlled delivery
of these substances. Some definitions explicitly restrict the external size of the
particles or their internal structures to no more than 100 nm. However, in the field
of agriculture, the accepted sizes range from 10 to 1000 nm [6]. Intelligent controlled
release systems for fertilizers, herbicides, and pesticides offer the possibility of
releasing the active substance at a specific time and place, minimizing nutrient losses
and optimally managing resources to increase crop yields [4, 7]. Its use avoids the
periodic application of agrochemicals and offers improvements regarding the safety
during handling these substances. In this sense, the benefits include greater effi-
ciency, increased yields, reduction of negative impacts on the environment and
health, and low costs [2, 5, 7]. In addition, controlled release systems have found
new agricultural applications that address the problem of maximizing production
from another approach. Such systems allow the delivery of phytoactive substances,
such as nucleotides, proteins, and genetic material, in order to directly modify and
regulate plant metabolism [7].
The design of the nanoparticles should be made conscientiously from the outset
considering the possible adverse effects on the environment and humans [7]. The use
of metallic nanoparticles such as Ag, Au, Cu, and oxidic nanoparticles (TiO2, ZnO,
CeO2, La2O3, etc.), among others, is a sensitive issue. These nanoparticles can cross
into plant tissues and organs [7]. Although many advantages of their uses have been
reported, they also might have serious problems when considering the putative
effects on ecosystems. For example, it has been reported that these nanoparticles
can cause toxicity by different pathways such as the dissolution and release of toxic
ionic species, the mechanical obstruction of certain pathways, the production of
reactive oxygen species, and the catalytic oxidation of biomolecules [7]. Moreover,
inorganic nanoparticles have been proved to be transferred to higher level organisms
fed with pastures treated with them, which is a worrying source of food contamina-
tion for humans [7].
In this scenario, polymeric materials, especially nanoparticles, represent a more
than attractive alternative. The use of plastics in agriculture began in the 1930s with
the use of greenhouse covers and agricultural mulches and has since been increasing.
4 D. Merino et al.

Nowadays, there is a great tendency in the use of natural and biodegradable poly-
mers, driven by the necessity of the replacement of the polymers derived from
petroleum. These polymers can be used in the manufacture of nanoparticles for the
slow release of agrochemicals or bioactive molecules. Another advantage of the use
of biodegradable polymers is that their degradation products can act as compost
improving soil quality and avoiding the accumulation of toxic waste of conventional
practices [8]. In turn, naturally occurring polymers are gaining considerable accep-
tance over synthetic polymers for these specific applications because of their eco-
logical nature, low cost, availability, and biodegradability [2].
In particular, this chapter highlights studies on polysaccharide-based systems as
controlled release for agrochemicals such as herbicides, pesticides, and fertilizers.
The most commonly used polysaccharides, chitosan and alginate, and the effects
produced by these polysaccharide-based nanoparticles in plants will be summarized.
Finally, updated information on the market availability of these systems for agricul-
tural applications and the existing regulation for this sector is presented.

Polysaccharides as Controlled Release Systems for Agrochemicals

Natural polymers play a very important role in the development of controlled release
systems for agrochemicals. Polysaccharides are the most suitable natural polymers to
meet these properties besides their wide availability, low cost, absence of toxicity,
biodegradability, and biocompatibility. Polysaccharides are polymeric carbohydrates
made up of units of monosaccharides linked together by glycosidic bonds. They are
highly versatile since they possess numerous reactive functional groups such as
hydroxyl, amino, and carboxyl, which can be used in chemical reactions to introduce
new groups in their structure [9]. These compounds are widely available in nature,
forming part of algae, plants, microorganisms, and animals. Their chemical struc-
tures are very diverse: linear, branched, or cyclic polysaccharides and some of them
can be neutral, while others have positive or negative charges [10]. Table 1 summa-
rizes chemical structures of main polysaccharides used for the formulation of
controlled release systems of agrochemicals and biomolecules with agricultural
applications.
According to the procedure used to obtain nanoparticles for the controlled release
of active agents and the characteristics of the agent to be conveyed, different types of
structures can be obtained (Fig. 1).
The nanocapsules are composed of a hydrophilic polymeric shell and a hydro-
phobic oily core which generally contains the active principle. Nanospheres are
polymeric aggregates where the active principle is homogeneously distributed in the
matrix and interacts with it through physical bonds [2, 12]. The following are the
main methods used to obtain these nanostructures:

Crosslinking polysaccharide nanoparticles: Crosslinking may be of chemical (cova-

lent) or physical (ionic) type. Covalent crosslinking forms chemical bonds and is
therefore irreversible. A rigid network that allows absorption of water and active
Table 1 Chemical structure, composition, and origin of the main polysaccharides commonly used in the formulation of nanoparticles for agricultural
applications
Polysaccharide Chemical structure Composition Source
Chitosan Monomers of N-acetyl-D- Derived from chitin, which is found
glucosamine and D-glucosamine in the exoskeleton of crustaceans [2]
linked by β(1!4) glycosidic bonds
[11]

Alginate Monomers of β-D-manuronic acid Brown macroalgae and bacteria [12]

and α-L-guluronic acid linked by
β(1!4) and α(1!4) bonds,
respectively [2]
Polysaccharides as Eco-Nanomaterials for Agricultural Applications

Cellulose Units of β-D-glucopyranose linked Cell wall of plants [2]

by β(1!4) glycosidic bonds [2]
5

(continued)
 6

Table 1 (continued)
Polysaccharide Chemical structure Composition Source
Starch Composed by two polymers: Plants, cereal grains, fruits, legumes,
amylopectin and amylose. The and roots [2]
former is branched and composed
by glucose units linked by α(1!4)
bonds and with branches linked by
β(1!4) bonds. Amylose is lineal
and is composed by glucose units
linked by α(1!4) bonds [2]
D. Merino et al.
Cyclodextrin Composed by α-D-glucopyranose
units linked by α(1!4) glycosidic
bonds [2]
Enzymatic hydrolysis of starch
molecules [13]
Polysaccharides as Eco-Nanomaterials for Agricultural Applications
7
8 D. Merino et al.

Fig. 1 Nanocapsules obtained by (a) ionic crosslinking, (b) covalent crosslinking, (c) complexa-
tion with a polyelectrolyte, and (d) self-assembled; nanospheres obtained by (e) ionic crosslinking,
(f) covalent crosslinking, and (g) complexation with a polyelectrolyte

compounds with high stability is formed. These particles are resistant in any pH
range. Glutaraldehyde is one of the most commonly used cross-linking agents,
but because of its toxicity, its application is limited and instead of it di- and
tricarboxylic acids such as maleic, tartaric, succinic, and citric acid are commonly
used [10, 11]. Physical crosslinking, on the other hand, is based on the electro-
static interactions between charged polysaccharides and their ionic crosslinking
agents. Ionic crosslinking has more advantages than covalent since it uses mild
and simple preparation conditions [10]. These nanoparticles are sensitive to
changes in pH, which can be used as a method to regulate the release of retained
substances. The most used crosslinking agent in this case is sodium tri-
polyphosphate mainly for the preparation of chitosan nanoparticles and encapsu-
lation of numerous active ingredients. Other crosslinking agents are citrates and
calcium ions which are generally used for the preparation of alginate nano-
particles [11]. The covalent or ionic crosslinking method then allows the forma-
tion of nanogels in the form of nanospheres that can retain and release a wide
range of substances (Fig. 1e, f) [10]. This technique can also be used for the
formation of nanocapsules as in Fig. 1a, b. For this purpose, generally an aqueous
solution of the polysaccharide is prepared and mixed with an organic solution of
the drug to be incorporated, and by sonication or agitation, oil-in-water type
emulsions are formed where the drug is surrounded by the polymer. Once the
emulsion is obtained, the physical or chemical cross-linking agent can be added
for the formation of the nanocapsules [9–11].
Polysaccharide nanoparticles by complexation of poly-ions: This method is similar
to the method of ionic crosslinking, with the difference that in this case poly-
electrolytes are used as complexing agents. Generally, chitosan is used as a
cationic polysaccharide and an anionic polysaccharide to form the complex
Polysaccharides as Eco-Nanomaterials for Agricultural Applications 9

[10]. The most commonly used anionic polysaccharides are carboxymethyl

cellulose, hyaluronic acid, and alginate. The complex formed has a structure as
outlined in Fig. 1g, and its stability depends on many variables, namely: solution
pH, ionic strength, temperature, molecular weight of the involved polyelectro-
lytes, and the flexibility of their chains. Another possibility is to use polypeptides
instead of polysaccharides to complex with chitosan [10, 11]. As with the
crosslinking method, polysaccharide nanocapsules can also be obtained by com-
plexation of polyelectrolyte polymers to the previously formed emulsion giving a
structure as shown in Fig. 1c [12].
Self-assembled polysaccharide nanoparticles: Since the polysaccharides have a
large number of reactive functional groups such as hydroxyl, carboxyl, or
amino groups, they can be chemically modified to introduce hydrophobic seg-
ments into their structure. These types of reactions are called “graft reactions”
[11]. Upon contact with water, the polymer molecules resulting from the modi-
fication are amphiphilic, and organized spontaneously by forming micelles or
aggregates by inter- or intramolecular associations of the hydrophobic moieties to
minimize the interfacial energy of the nanoparticles [10]. The self-assembly
depends on factors such as the molar ratio between the polysaccharide and the
hydrophobic segments and the length of the latter. In this way, those drugs that are
insoluble in water can be encapsulated in a hydrophobic core and become soluble
in water, thanks to the external hydrophilic layer. The release is then susceptible
to changes in pH, ionic strength, temperature, and other external stimuli [11].

The above-described structures are often used for transport and release of herbi-
cides, pesticides, and fertilizers in a controlled manner. Nanoherbicides are nano-
particles that house and release herbicidal substances. Their function is to prevent
weed growth as they significantly reduce crop strength and yield. Conventionally,
the herbicides are applied directly on the aerial part of the weeds, so that their roots
remain unaffected and after a time reappear. The application of nanoherbicides in the
soil and their slow release provides small doses that control weed growth, avoiding
potential adverse effects on crops [4–14]. Fertilizer nanoparticles are used to
improve the efficiency of the use of nutrients since these are released depending
on the requirements of plants, thus avoiding premature conversion of these into
chemical forms inaccessible to the plant [7]. Nitrogen is one of the most important
nutrients in fertilizers and the efficiency in its use is currently very low. Reducing the
efficiency of fertilizers increases crop costs. The indiscriminate use of fertilizers can
contaminate natural resources such as soil, water, and air, creating serious dangers
for the entire ecosystem. Nanopesticides are release systems of chemicals commonly
used for pest control. Conventionally, these are added directly causing great con-
tamination of soil and water and generating high costs to producers [4]. Usually,
these active ingredients are poorly soluble in water, harmful to nontarget organisms,
and produce resistance in target ones. In turn, nanoencapsulation helps to make their
management and administration safe and reduces current pollution levels [5]. Addi-
tionally, nanoparticles have also been employed for the controlled release within the
plant cells of molecules with biological activity such as genetic material and elicitors
10 D. Merino et al.

of gene expression among others. This particular strategy would induce gene
expression without integrating plasmid DNA with the plant genome and thus
avoid the transfer of the changes introduced to the next generation [5, 11]. This
technique is highly efficient since it uses up to 1000 times less DNA than traditional
techniques and allows obtaining “nontransgenic” modified plants [4, 7].

Chitosan and Alginate-Based Nanomaterials for Promoting Plant

Growth and Protection Against Stresses

Chitosan and alginate have been the most widely studied biopolymers. Chitosan is a
linear cationic polymer and is obtained by the partial deacetylation of chitin. Chitin is
found extensively in nature in the cell walls of certain fungi and bacteria and forming
part of the exoskeletons of marine crustaceans. Meanwhile, alginate is majority
obtained from brown macroalgae and bacteria [2, 12]. Chitosan is one of the most
abundant renewable resources on earth. Chitosan is notable for its chemical and
biological properties. It has been widely studied in biomedical, foods, agriculture,
engineering, and environmental sciences [11]. It is a hydrophilic polymer with low
solubility in water, so it is generally dissolved in mild acid medium by protonation of
its amino groups [11]. Chitosan is consumer-safe, nonallergenic, and biocompatible
compound [15]. On the other hand, alginate is an anionic linear polysaccharide,
soluble in water with high availability, biodegradability, biocompatibility, and
absence of toxicity [11, 12]. These two polymers, chitosan and alginate, are widely
used for the formation of particles called hydrogels. Hydrogels are three-dimensional
crosslinked polymer networks with high water absorption and swelling capacity
[16]. In particular, nanogels are polymeric nanoparticles of hydrogels. They have
greater mechanical strength and surface area than micro or macrogels and thanks to
their nanometric size they can cross cell nozzles and channels [12].
The single alginate as matrix to prepare controlled delivery compounds has
inherent poor mechanical strength and uncontrolled degradation resulting in
unpredicted release rates of the bioactive. For these reasons, these traits are
counteracted by blending with other polymers such as chitosan, gelatin, or
carboxymethyl cellulose [17, 18].
In particular, chitosans are able to interact with different compounds since they
are linear copolymers of acetylated and nonacetylated residues. This fact yields free
amino groups which at slightly acidic pH values convey positive charges to chitosan,
making it the only polycationic polysaccharide. This property confers to them
remarkable properties and biological functionalities including electrostatic interac-
tions with polyanionic molecules such as proteins, nucleic acids, polyanionic
phospholipidic, and sulfated polysaccharides [19]. Traditional approaches for pro-
ducing chitosans have used physical and chemical means inevitably leading to broad
mixtures of polysaccharide containing a variety of degrees of polymerization and
acetylation. It has been summarized that chitosan production has crossed at least
three periods: (i) decades ago agro-chitosans have implied weakly defined polymeric
mixtures with varied purity and composition, (ii) then, chitosans have represented
Polysaccharides as Eco-Nanomaterials for Agricultural Applications 11

mixtures chemically better characterized in terms of degrees of polymerization and

acetylation; and (iii) the last chitosan generation includes novel polysaccharides
biotechnologically synthesized in heterologous expression systems [20–22].
In agriculture, the first two periods of chitosans have conferred a wide framework
of applications as plant growth promoters and seed priming [23, 24], in addition to
improve tolerance to abiotic such as temperature, salinity, and drought [25] and
biotic stresses including, virus, bacteria, fungi, and insects [26–37]. However, in
addition to the low water solubility at neutral pH, solution of chitosans weakly
characterized has conducted to a rather reproducibility in terms of biological effects.
Likely, these two traits have been partly responsible for their nonexpansive applica-
tion in agriculture. An alternative approach to counter the solubility problem has
been to use chitooligosaccharides rather than polymers [38, 39]. Research has
elucidated detailed molecular structure-function relationships of chitosan in a high
number of plant species including mono- and dicotyledonous. More recently, the
design of chitosan-based nanomateriales which can be dispersed in aqueous solution
have represented new opportunities to enlarge its applications in agroindustry. Major
advantages of encapsulating agrochemicals in chitosan nanoparticles include con-
trolled release, stability, and protection of active ingredients from the surrounding
environment allowing them to serve as efficient bioactive delivery systems [40]. In
the same direction with the previous summarized background for solutions of
chitosans, chitosan-based nanoparticles prepared from different carbon sources
might evidence different structure-functional relationships [15]. Either way, the
efficiency or efficacy of chitosan-based nanoformulations will be valued according
to higher surface area, lower toxicity, and induction of desired biological activity
[41]. Indeed, selected procedure for the synthesis, strategies for loading bioactive
compound into nanoparticles, type and molecular mass of chitosan, concentration,
and ionic strength, among other parameters, can affect the efficacy of chitosan-based
systems [15, 40].
Compared with synthesis and action of metal nanoparticles with action in plants,
there is much lower number of studies that refer to chitosan-based nanoparticles [42,
43]. In summary, supporting by their associated properties and biological functions,
chitosans have been proposed as useful polymers for the efficient delivery of
agrochemicals, including hormones, pesticides, and herbicides with lower toxicity
and genotoxicity than free compounds [44–46], and, in turn, allowing new market
opportunities in the agricultural field [17, 45, 47]. Chitosan-based nanoparticles have
also demonstrated positive action on germination promoting and stimulation of plant
growth [48]. Other example includes chitosan nanoparticles with antifungal activity
resulting in promissory biological pesticides [49, 50].
Novelty in the study on the immunomodulatory role of chitosan-based nano-
particles in plants has been lately initiated [51]. Today, polysaccharides and free
sugars are considered as signaling molecules rather than just carbon sources and
energy in plants. Both types of molecules would compromise what are now
known as classic and sweet immunities [52, 53]. However, it remains unclear
about the receptors and molecular mechanisms that plant utilizes to recognize and
transduces each poly and monosaccharide signals [54–59]. In addition to the nature
12 D. Merino et al.

of carbohydrate molecules, size, acetylation, and concentration can determine the

intensity and temporality of plant defense responses [60]. The diversity of responses
observed in a variety of plant species probably shows the complexity of the
perception and transduction mediated by mono-, oligo-, and polysaccharides.
Another important aspect to clarify involves the way the materials are absorbed
and translocated through the plant cells and tissues. Likely, the accumulation of
antioxidant compounds, the participation of nitric oxide as a signaling molecule, and
activation of defense-related gene expression are common components among the
mechanisms of action described for both free chitosan and chitosan-based nano-
particles [23, 51, 61–68]. At cellular level, oligo- and polysaccharides have been
shown to induce various types of responses, including transient depolarization of the
membrane, generation of hormones, and induction of specific defense genes, all of
them integrated in complex and uncompleted deciphered networks [68–72]. Hence,
the picture that might emerge from the study on mechanism of actions mediated by
these types of biomaterials could be complex. For all these reasons, it is still an active
area of research but evidencing that most of these cellular events can also be
associated to the action of chitosan-based nanomaterials. In the near future, the
integration of smart systems for plant growth promotion, food processing, and
nano-based technologies to characterize plant gene expression could ensure more
successful sustainable food production.

Legal Aspects and Current Regulation on Nanomaterials Usage

Unique physiochemical properties of nanoparticles make the investigation of their

toxic consequences intricate and challenging, so that the knowledge of different
mechanisms involved in nanomaterials’s action as well as toxicity is required.
Nanotoxicity has various effects on human health and diseases as they can
easily enter into the humans via different routes: mainly respiratory, dermal, and
gastrointestinalones.
Nanoparticles toxicity is very complex and multifaceted as it depends on a variety
of physicochemical and surfacial properties like their size, shape, charge, area, and
reactivity. The nanoparticles (<100 nm) are more toxic than larger particles of
identical chemical composition [73].
A systematic understanding of the nanomaterials interactions with biological
systems at cellular, molecular, and physiological levels is essential for understanding
the possible unsafe responses. The exposure to nanoparticles adversely affect mam-
mals and other species at cellular, organ, and tissue level by causing oxidative stress
and inflammation. It also leads to the altered function of the autonomic nervous
system that in turn results in enhanced respiratory and cardiovascular diseases. NPs
can enter the blood circulation and migrate to different organs and tissues, and injure
oxidative stress-sensitive organs. In general, the toxicology related with nano-
systems affects lungs functioning, alters heart rate and blood pressure, and displays
respiratory symptoms, thrombosis, myocardial infarction, arrhythmia, and strokes
causing shorter life expectancy [74] (Fig. 2).
Polysaccharides as Eco-Nanomaterials for Agricultural Applications 13

• Reduce Risk • Exposure

• Physicichemical Properties • Biodegradation
• Studies (in vitro and in vivo) • Interaction with
• Duration ecosystem
• Dosis controlled release
•Specific'site release Products Final
•Release on demand
based on deposition in
nanoparticles the
design environment

Risk
Regulation
Estimation

•Poor or absent Availability • Experimental Toxicity Data

•Yes: Healt Risk? • Modelling of toxicity
•No: Formulation guidelines • Real conditions to test

Fig. 2 Scheme of considerations related with risk associated with nanoparticle-based products

Nanotechnology Risk Assessment and Regulation in the EU and

Worldwide

Due to the variety of applications of nanotechnology, different parts of legislation are

concerned in the European Union, including horizontal legislation and product-
specific legislation. The most comprehensive legislation relevant to nanomaterials
is the EU Regulation on Registration, Evaluation, Authorization and Restriction of
Chemicals (REACH), which addresses chemical substances, in whatever size, shape,
or physical state. Substances at the nanoscale are therefore covered by REACH, and
its provisions apply.
Among product-specific legislation, some already explicitly address nano-
materials (cosmetics, foods additives, provision of food information to consumers,
and biocides) while others do not (for example: toys, electrical equipment, and waste
environmental legislation). At international level, there are several activities in place
on risk analysis of nanomaterials in the food and agricultural sectors. Overall,
definitions of nanomaterials developed in different countries result in different risk
management measures. So far, apart from the EU, no country has set a regulatory
framework for the mandatory labeling of nanomaterials.
14 D. Merino et al.

Table 2 Regulation regarding nanomaterials

Document References
European Parliament 2006 – Nanosciences and nanotechnologies: an action plan [79]
for Europe 2005–2009
European Parliament 2009 – Regulatory aspects of nanomaterials [80, 81]
European Commission 2004 – Towards a European strategy for nanotechnology [82]
European Commission 2005 – Nanosciences and nanotechnologies: an action plan [83]
for Europe 2005–2009
European Commission 2007 – Nanosciences and nanotechnologies: an action plan [81]
for Europe 2005–2009. First implementation report 2005–2007
European Commission 2008 – Code of conduct for responsible nanosciences and [84]
nanotechnologies research
European Commission 2011 – Definition of nanomaterial [85]
EFSA 2015 annual report of the EFSA scientific network of risk assessment of [86]
nanotechnologies in food and Feed1 for 2014

Regulatory Issues

Most of the countries do not have defined regulations for the marketing and use of
nano products. European policy is among those which tried to establish a strong and
defined regulation. Although the existing laws were considered for conventional
food products, the same laws have been also considered on a broad aspect of nano-
foods. In the last decade, the European regulatory debate has been characterized by a
change of perspective notably supported by the European Parliament, which in 2009
required that the Commission review all relevant legislation within 2 years to ensure
that legislative provisions and instruments of implementation reflect the particular
features of nanomaterials to which workers, consumers, and/or the environment may
be exposed – European Parliament Resolution on regulatory aspects of nano-
materials [75]. Consequently, the vast need of some regulations containing specific
provisions addressing nanomaterials have made the entrance of laws/regulations.
However, specific regulations do not exist for all food and agro categories; therefore,
it is quite difficult for food industry and private sectors to have clear guidance on the
applicable regulatory framework [76]. Literature and earlier studies on nano-food
regulation identifies the incapability of the existing regulatory processes and keep a
rapid policymaking and regulations that are required for entering the marketplace
[77, 78]. Regulations related with nanomaterials are summarized in Table 2.

Current Nanomaterials in the Agricultural Field (Commercial and

in Developing)

The application of nanomaterials in agriculture aims in particular to reduce applica-

tions of plant protection products, minimize nutrient losses in fertilization, and
increase yields through optimized nutrient management. Despite these potential
advantages, the agricultural sector is still comparably marginal and has not yet
made it to the market to any larger extent in comparison with other sectors of
Polysaccharides as Eco-Nanomaterials for Agricultural Applications 15

nanotechnology application. Nanotechnology devices and tools, like nanocapsules,

nanoparticles, and even viral capsids, are examples of uses for the detection and
treatment of diseases, the enhancement of nutrients absorption by plants, and the
delivery of active ingredients to specific sites and water treatment processes. The use
of target-specific nanoparticles can reduce the damage to nontarget plant tissues and
the amount of chemicals released into the environment. Nanotechnology-derived
devices are also explored in the field of plant breeding and genetic transformation.
The potential of nanotechnology in agriculture is large, but a few issues are still to
be addressed, such as increasing the scale of production processes and lowering
costs, as well as risk assessment issues. In this respect, particularly attractive are
nanoparticles derived from biopolymers such as proteins and carbohydrates with low
impact on human health and the environment. For instance, the potential of starch-
based nanoparticles as nontoxic and sustainable delivery systems for agrochemicals
and bioestimulants is being extensively investigated.
Nanomaterials and nanostructures with unique chemical, physical, and mechan-
ical properties – e.g., electrochemically active carbon nanotubes, nanofibers, and
fullerenes – have been recently developed and applied for highly sensitive biochem-
ical sensors. These nanosensors have also relevant implications for application in
agriculture, in particular for soil analysis, easy biochemical sensing and control,
water management, and delivery, pesticide and nutrient delivery.
From a commercial perspective, existing agrochemical companies are investigat-
ing the potential of nanotechnologies and, in particular, whether intentionally
manufactured nano-size active ingredients can give increased efficacy or greater
penetration of useful components in plants. However, the nano-size so far did not
demonstrate to hold key improvements in product’s characteristics, especially con-
sidering the interest of large-scale production and the costs involved in it.
Some specific nano-products for the agricultural sector have been put on the
market by technology-oriented smaller companies, like soil-enhancer products that
promote even water distribution, storage, and consequently water saving. However,
the commercial market application of these products is so far only achieved at small
scale, due to the high costs involved in their development. These costs are normally
compensated by higher returns in the medical or pharmaceutical sectors, but so far
there are no such returns in the agricultural sector. Research continues in the
commercial agrochemical sector to evaluate potential future advantages.
Companies are also facing challenges derived from the definition of nano-
materials that is adopted by the EU. One crucial point related to the EU definition
is the possibility that nonactive substances already used for many decades in
commercial products formulations will fall within the scope of the nano definition,
although not intentionally developed as nanoparticles or having specific nanoscale
properties. Nanoscale formulants (e.g., clay, silica, polymers, pigments, macromol-
ecules) have been used for many decades and are also ubiquitous in many daily
household products.
The concern is that the need for labeling of products that are already on the
market since decades results in a scenario, in which the technology is stigma-
tized, preventing further and innovative applications of nanotechnology in agricul-
ture (Table 3).
16
Table 3 Some applications of nanotechnology in agriculture
Application
Fertilizers: Nanocapsules,
nanoparticles, and viral capsids for the
enhancement of nutrients absorption by
plants and the delivery of nutrients to
specific sites
Nanocides: Pesticides encapsulated in
nanoparticles for controlled release.
Nanoemulsions for greater efficiency
Nanosensors and diagnostic devices:
Nanomaterials and nanostructures (e.g.,
electrochemically active carbon
nanotubes, nanofibers, and fullerenes)
that are highly sensitive biochemical
sensors to closely monitor
environmental conditions, plant health,
and growth
Nanosensors: Contamination of
packaged food. Pathogen detection
Buckyball fertilizer: Ammonia from
buck balls
Water/liquid retention:
Nanomaterials, e.g., zeolites and nano-
clays, for water or liquid agrochemicals
retention in the soil for their slow
release to the plants
Plant protection products:
Nanocapsules, nanoparticles,
nanoemulsions, and viral capsids as
smart delivery systems of active
ingredients for disease and pest control
in plants
Water purification and pollutant
remediation: Nanomaterials, e.g.,
nano-clays, filtering, and binding to a
variety of toxic substances, including
pesticides to be removed from the
environment
Plant genetic modification:
Nanoparticles carrying DNA or RNA
to be delivered to plant cells for their
genetic transformation or to trigger
defense responses against pathogens
D. Merino et al.
Example References
Macronutrient fertilizers coated with [87]
zinc oxide nanoparticles (University of
Adelaide, AUCSIRO Land and Water,
AU Kansas State University, USA)
BASF, Ludwigshafen, Germany [88]
Pesticide detection with a Liposome- [89]
based nano-biosensor (University of
Crete, GR)
Nestle, Kraft, Chicago, USA; Cornell [90, 91]
University, Vevey, Switzerland
Kyoto University, Kyoto, Japan [92]
Soil-enhancer product, based on a [93]
nano-clay component, for water
retention and release (Geohumus-
Frankfurt, DE)
Neem oil (Azadirachta indica) [94]
nanoemulsion as larvicidal agent (VIT
University, IN)
Filters coated with TiO2 nanoparticles [95]
for the photocatalytic degradation of
agrochemicals in contaminated waters
(University of Ulster, UK)
Mesoporous silica nanoparticles [96]
transporting DNA to transform plant
cells (Iowa State university, USA)
Polysaccharides as Eco-Nanomaterials for Agricultural Applications 17

Conclusions and Perspectives

The use of polysaccharide-based nanoparticles as smart delivery system in the

agriculture field is still in its infancy. Significant outcomes have been reported
on plant growth promotion and stress protection by chitosan and alginate-
chitosan–based nanomaterials at the laboratory and green house scales. Since poly-
saccharide-based nanomateriales have evidenced highly successful results for the
controlled delivery of different agrocompounds (pesticides, plant growth regulators,
herbicides, nutrients) at the laboratory scale, each of them would have great potential
to be applied in the field in the near future. Chitosan-based nanomaterials could be
the most promising polysaccharides for the development of controlled release
systems for agricultural application. The combination and vehiculization of more
than one bioactive (i.e., two herbicides or herbicide plus fungicide) and matrix
(chitosan/alginate) might be potential and enriches the functionality of a defined
chitosan-based nanomaterial. In spite of all the previously mentioned advantages, the
scarce and nascent information on the chitosan-based nanoparticules still makes it
necessary to raise caution about the potential use in the natural environment. In order
to improve our understanding and assessment of chitosan-based nanomaterials, the
following issues have yet to be deepened:

– Specificity versus universality of their biological properties

– Biophysical and chemical interactions with other components abundant in plants
and the environment where they inhabit
– Mechanisms of action on specific cells, tissues, organs, and plants
– Interactions with biota they interact
– Life cycle assessments

Other important point to remark is that several countries over the world have been
active in examining the appropriateness of their regulatory frameworks for dealing
with nanotechnologies. Collaboration among countries worldwide is required in
order to exchange information and to ensure a high level of protection for humans
and the environment, while not hampering the development of new beneficial
products and their global marketing.

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