NANOSYSTEMS BASED ON NANOCOMPOSITES AND NATURAL EXTRACTS

Abstract

The instant invention refers to nanosystems comprising nanocomposites for adsorption or support of natural extracts; a process for the preparation thereof; formulations containing thereof, as well as a nanomaterial that adsorbs one or more essential oils in its surface. Specially, one object of the invention is the encapsulation of natural extracts, i.e., essential oils and/or natural pure bioactive compounds and optionally terpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, carotenoid, and ricinoid compounds; complementary acids, and polysaccharides; vitamins, and other organic compounds, in mesoporous materials for application in different industries such as food, agricultural, veterinary, aquacultural, pharmaceutical, cosmetic, cleaning, sanitizing, and disinfection industries, as well as in medicine.

Claims

1-90. (canceled)

91. A nanosystem comprising a mesoporous material support formed from a nanocomposite and natural extracts deposited and/or adsorbed on the surface of said mesoporous material wherein the nanocomposite comprises titanium dioxide (TiO2)-MO being M a transition metal which is not considered toxic for environment, mammals, and plants, having the nanocomposite a surface area from about 74 m.sup.2/g to 200 m.sup.2/g and wherein the concentration of natural extracts is from about 10 mg to 100 mg and wherein the morphology of the nanosystem is nanospheres or nanodrops.

92. The nanosystem of claim 91, wherein the nanocomposite is based on TiO.sub.2ZnO.

93. The nanosystem of claim 91, wherein the natural extracts are encapsulated by terpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, carotenoid, resinoids, complementary acids and polysaccharides, vitamins and other organic compounds.

94. The nanosystem of claim 91, wherein the natural extracts are essential oils and/or fruit bioactive compounds alone or combined with two or more thereof, obtained from flowers selected from Arnica montana, Lavandula sp., Chamaemelum nobile, Tanacetum cinerariifolium, Thymus sp., Syzygium aromaticum, Rosa sp., Geranium sp., Jasminum sp., Cananga odorata, Citrus aurantium var. Amara, Lavandula sp., Plumeria rubra, Borago officinalis, Erodium cicutarium, Gnaphalium sp., Heterotheca inuloides Cass., Lepidium virginicum L., Matricaria recutita L., Mirablis jalapa L., Tagetes lucida Cav; roots selected from Angelica archangelica, Asarum europaeum, Crocus sativus, Acorns calamus, Curcuma longa, Alpinia galanga, Zingiber officinale, Santalum album, Sassafras albidium, Valeriana officinalis, Chrysopogon zizanioides, Glycyrrhiza glabra L., Cinnamoum verum, Agave spp.; leaves selected from Artemisia absinthium, Ocimum basilicum, Agathosma betulina, Aloysia citrodora, Eucalyptus sp., Mentha spicata, Cymbopogon, citratus, Origanum majorana, Mentha sp., Pogostemon cablin, Chenopodium ambrosioides, Salvia rosmarinus, Salvia officinalis, Melissa officinalis, Cinnamomum verum, Moringa oleifera, Organum vulgare L., Plantago major L., Taraxacum officinale W.; the fruit pericarp selected from Citrus bergamia, Citrus x limon, Citrus reticulata, Citrus x sinensis, Citrus x aurantium, Citrus x latifolia, Citrus x paradisi, Agave spp., Juglans regia L., Punica granatum L.; seeds selected from Pimpinella anisum, Elettaria cardamomum, Morinda citrifolia, Anethum graveolens, Foeniculum vulgare, Cuminum cyminum, Salvia officinalis, Salvia hispanica, Capsicum annauum, Rosa rubiginosa, Vitis vinifera, Cocos nucifera, Argemone mexicana L., Avena sativa; fruits selected from Carum carvi, Coriandrum sativum, Laurus nobilis, Myristica fragans, Petroselinum crispum, Piper nigrum, Morinda citrifolia; stems or branches selected from Cinnamomum verum, Cedrus sp., Pinus sp., Eucalyptus sp., Abies sp., Cupressus sp., Agave spp., Aloe barbadenses Mil.

95. The nanosystem of claim 94, wherein the essential oils are selected from anethol, anisaldehyde, bomeol, carvacrol, D-carvone, I-carvone citral citronellal, geraniol, D-limonene, linalool, menthol, pinene, terpineol, thymol, vanillin, alfa-ocimene, borneol, Y-cadinene, caryophyllene, citronellal, p-cymene, aldehyde decyilic, farnesol, farnesal, fenchone, geraniol, geranyl acetate, germacrene, limonene, methyl heptenone, myrcene, nerolinol, nerol, ocimene, terpinene, -pinene, -phellandrene, -myrcene, -terpinolene, octanal, decanal, octanol, iso-citronellene, camphene, trans-p-menthane, p-mentha-1(7),8-diene, dihydromyrcenol, trans-dihydrocarvone, alpha-pinene, beta-pinene, estragole, longifolene, L-alpha-terpineol, alone or combined with two or more thereof.

96. The nanosystem of claim 91, which further optionally comprises terpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, carotenoid, and ricinoid compounds; complementary acids selected from lactic acid, palmitic acid, formic acid, citric acid, oxalic acid, ureic acid, ascorbic acid, malic acid, acetic acid, alone or combined with two or more thereof, complementary polysaccharides selected from glucose, ribose, deoxyribose, mannose, fructose, galactose, glyceraldehyde, erythrose, fucose, alone or combined with two or more thereof; vitamins selected from vitamin A, thiamin B1, riboflavin B2, niacinamide B3, pyridoxin B6, cobalamin B12, vitamin D, vitamin C, vitamin E, folic acid (vitamin B9), pantothenic acid (vitamin B5 or W), alone or combined with two or more thereof; and other organic compounds selected from bioflavonoids, glycerin, pectins, and amino acids, alone or combined with two or more thereof.

97. The nanosystem of claim 91 for use as a biocide.

98. A vermicide formulation comprising the nanosystem of claim 91.

99. An antifungal formulation comprising the nanosystem of claim 91.

100. A bactericidal formulation comprising the nanosystem of claim 91.

101. A pesticidal formulation comprising the nanosystem of claim 91.

102. A disinfectant formulation comprising the nanosystem of claim 91.

103. A sanitizing formulation comprising the nanosystem of claim 91.

104. A germicidal formulation comprising the nanosystem of claim 91.

105. A virucidal formulation comprising the nanosystem of claim 91.

106. A process for obtaining the nanosystem of claim 91 comprising a mesoporous material support formed from a nanocomposite and one or more natural extracts deposited and/or adsorbed on the surface of said mesoporous material, the process comprising the following steps: a) to obtain natural extracts by drying, milling, and sieving peels and seeds of fruits, and then carrying out a metabolic extraction by ultrasound-assisted sonication, b) to prepare the nanocomposite by the sol-gel method by mixing titanium dioxide (IV), polyethylene glycol (PEG) and ethanol, to obtain a solution; c) to heat the solution from step (b) between 60 and 120 C. under reflux; d) to add distilled water containing Zn(NO.sub.3).Math.6H.sub.2O in the preparation of each one of the series to obtain materials with 1.0, 3.0, 5.0, and 10.0% by weight; e) to add to the obtained solution of step (d) of the corresponding metallic salt, a few drops of HNO.sub.3 until a pH value of about 3 in said solution; f) to add dropwise to the solution of step (e), butoxide-ethanol having a molar rate butanol/water of 8:1; g) to mix the solution of step (f) under magnetic stirring until a gel is formed; h) to cool the solution of step (g) to about 0 C. and live it stand at about 4 C. and then dry the gel at about 100 C.; and i) to mill the solid obtained in step (h), and then calcined it at a temperature between 400 and 600 C. under an air atmosphere, and mill it again.

107. The process of claim 106 which further comprises to add in step (a) terpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, carotenoid, and ricinoid compounds; complementary acids, and polysaccharides; vitamins, and other organic compounds.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0076] FIG. 1 illustrates the adsorption-desorption isotherms for the mixed oxides of TiO.sub.2ZnO (a: 1.0% by weight of Zn, b: 3.0% by weight of Zn, c: 5.0% by weight of Zn, d: 10.0% by weight of Zn).

[0077] FIG. 2 shows the X-ray diffractograms for the mixed oxides of TiO.sub.2ZnO (a: TiO.sub.2, b: 1.0% by weight of Zn, c: 3.0% by weight of Zn, d: 5.0% by weight of Zn, and e: 10.0% by weight of Zn).

[0078] FIG. 3 shows the Raman spectra of TiO.sub.2ZnO (a: TiO.sub.2, b: 1.0% by weight of Zn, c: 3.0% by weight of Zn, d: 5.0% by weight of Zn and e: 10.0% by weight of Zn).

[0079] FIG. 4 shows the UV-Vis spectra of the TiO.sub.2ZnO semiconductors (a: TiO.sub.2, b: 1.0% by weight of Zn, c: 3.0% by weight of Zn, d: 5.0% by weight of Zn and e: 10.0% by weight of Zn).

[0080] FIGS. 5a-5d show a Scanning Electron microscopy of TiO.sub.2ZnO a: 1.0% Zn, b: 3.0% Zn, c: 5.0% Zn and d: 10.0% Zn).

[0081] FIGS. 6a-6d show Transmission Electron Microscopy (TEM) of TiO.sub.2ZnO a: 1.0% Zn, b: 3.0% Zn, c: 5.0% Zn, d: 10.0% Zn.

[0082] FIGS. 7a-7c illustrate the evaluation of 7 ATCC reference strains and 2 control strains: a) in nutritive agar medium without extract; b) nutritive agar with 100% plant extract, c) nutritive agar with 50% plant extract.

[0083] FIG. 8 is a photomicrograph using scanning electron microscopy (SEM).

DETAILED DESCRIPTION OF THE INVENTION

[0084] The embodiments described in the present invention should in no way be construed as limiting thereof, but rather should be construed as illustrative, exemplifying the principles thereof. Any of the section titles used herein have organizational purposes only and should not be construed as limiting the subject matter disclosed.

[0085] In recent years, composites or mixed oxides have become increasingly interesting biomaterials, which constitute a new perspective in drug delivery systems and nanocarrier formulations due to their beneficial properties, including biocompatibility, biodegradability and low toxicity. The potentiality of chemical modifications of a biomaterial (mixed oxide), as well as its complementary use with other nanomaterials, attracts even more the scientific community, offering improved and combined properties in the end materials. Consequently, the present invention develops nanosystems based on a TiO.sub.2 nanomaterial and/or preferably a mesoporous TiO.sub.2-MO (M=Transition Metals) nanocomposite used as a matrix for the encapsulation and transport of several valuable compounds. The present invention describes as an example the use of a nanocomposite or mixed oxides based on TiO.sub.2ZnO for the production of nanosystems, without limiting the invention, focusing on the encapsulation of natural products for the encapsulation preferably of natural extracts such as essential oils and/or bioactive compounds.

[0086] In addition to essential oils, terpeneoids, sesquiterpenoids, diterpenoids, sesterpenoids, triterpenoids, carotenoids, resinoids, complementary acids and polysaccharides, vitamins and other organic compounds may be optionally added to the nanosystem.

[0087] The essential oils may be selected from anaetol, anisaldehyde, bomeol, carvacrol, D-carvone, 1-carvone citral citronellal, geraniol, D-limonene, linalol, menthol, pinene, terpineol thymol, vanillin, alpha-ocimene, borneol, Y-cadinene, caryophyllene, citronellal, P-cymene, decyl aldehyde, farnesol, farnesal, fenchone, geraniol, geranyl acetate, germacrene, limonene, methyl, heptenone, myrcene, nerolinol, nerol, ocimene, terpinene, -pinene, -phelandrene, -myrcene, -terpinolene, octanal, decanal, octanol, isocitronelene, camphene, trans-p-menthane, p-mentha-1(7), 8-diene, dihydromyrcenol, trans-dihydrocarvone, alpha-pinene, beta-pinene, estragole, longifolene, L-alpha-terpinol and can be used individually or two or more of them or in combination thereof.

[0088] The complementary acids may be selected from lactic acid, palmitic acid, formic acid, citric acid, oxalic acid, uric acid, ascorbic acid, malic acid, acetic acid and can be used individually or two or more of them or in combination thereof.

[0089] The complementary polysaccharides may be selected from glucose, ribose, deoxyribose, mannose, fructose, galactose, glyceraldehyde, erythrose, fucose and can be used individually or two or more of them or in combination thereof.

[0090] The vitamins may be selected from vitamin A, thiamine B1, riboflavine B2, niacinamide B3, pyridoxin B6, cobalamin B12, vitamin D, vitamin C, vitamin E, folic acid (vitamin B9), pantothenic acid (vitamin B5 or W) and can be used individually or two or more of them or in combination thereof.

[0091] Other organic compounds may be selected from bioflanoids, glycerin, pectins and amino acids and can be used alone or combined with two or more thereof.

[0092] Through the process of the present invention, mesoporous nanostructures are obtained, such as mesoporous nanospheres that have a high surface area, low density and good surface permeability due to the fact that it is a type of core-shell material with a special structure, the inner cavity can accommodate a large number of guest molecules, that is, it can achieve a high loading capacity of natural or synthetic substances, and the porous layer can be used as a channel for the release of natural or synthetic substances, that is, they have a slow release yield by adjusting the thickness of the coating, the pore size, the morphology of said pores and modification of their surface.

Definitions

[0093] Unless otherwise specified, a nanosystem will be understood herein as one comprising or being formed by one or more active ingredients (active substance(s) or molecule(s)) and a carrier system that can direct the release of the substance to a place or target in a lower and more effective dose.

[0094] In the present invention the term fruits shall be understood as the ovary of any plant species from the plant kingdom, developed after fertilization of the ovules, which will form the seed, regardless of the type of fruit, for example dried fruit or fleshy fruit. In addition to the ovary, some attached parts such as the receptacle, calyx, corolla, bracts, inflorescence axes, etc. can integrate the fruit.

[0095] Essential oils in the present invention shall be understood as volatile liquid fractions and which are complex mixtures of up to more than 100 components, such as low molecular weight aliphatic compounds (alkanes, alcohols, aldehydes, ketones, esters and acids), monoterpenes, sesquiterpenes, diterpenes, sesterpenes, triterpenes, carotenoids, resinoids, phenylpropanes, anaetol, anisaldehyde, bomeol, carvacrol, D-carvone, 1-carbona citral citronellal, geraniol, D-limonene, linalol, menthol, pinene, terpineol thymol, vanillin, alpha-ocimene, borneol, Y-cadinene, caryophyllene, citronellal, ID-cymene, decyl aldehyde, farnesol, farnesal, fenchone, geraniol, geranyl acetate, germacrene, limonene, methyl, heptenone, myrcene, nerolinol, nerol, ocimene, terpinene, -pinene, -phelandrene, -myrcene, -terpinolene, octanal, decanal, octanol, isocitronlene, camphene, trans-p-menthane, p-mentha-1(7), 8-diene, dihydromyrcenol, trans-dihydrocarvone, alpha-pinene, beta-pinene, estragole, longifolene, L-alpha-terpinol. However, the term essential oils could also be understood according to the context described, as established by the International Organization for Standardization (ISO), which defines them as the product obtained from a natural plant raw material, by steam distillation, by mechanical processes of the epicarp of citrus fruits, or by dry distillation, after separation of the aqueous phase, if any, by physical processes.

[0096] The essential oils can be isolated from different parts of the plant, for example from the flowers of Arnica montana, Lavandula sp., Chamaemelum nobile, Tanacetum cinerariifolium, Thymus sp., Syzygium aromaticum, Rosa sp., Geranium sp., Jasminum sp., Cananga odorata, Citrus aurantium var. Amara, Lavandula sp., Plumeria rubra, Borago officinalis, Erodium cicutarium, Gnaphalium sp., Heterotheca inuloides Cass., Lepidium virginicum L., Matricaria recutita L., Mirablis jalapa L., Tagetes lucida Cali; from the roots of Angelica archangelica, Asarum europaeum, Crocus sativus, Acorus calamus, Curcuma longa, Alpinia galanga, Zingiber officinale, Santalum album, Sassafras albidium, Valeriana officinalis, Chrysopogon zizanioides, Glycyrrhiza glabra L., Cinnamoum verum, Agave spp.; the leaves of Artemisia absinthium, Ocimum basilicum, Agathosma betulina, Aloysia citrodora, Eucalyptus sp., Mentha spicata, Cymbopogon, citratus, Origanum majorana, Mentha sp., Pogostemon cablin, Chenopodium ambrosioides, Salvia rosmarinus, Salvia officinalis, Melissa officinalis, Cinnamomun verum, Moringa oleifera, Organum vulgare L., Plantago major L., Taraxacum officinale W.; the pericarp of the fruit of Citrus bergamia, Citrus x lemon, Citrus reticulata, Citrus x sinensis, Citrus x aurantium, Citrus x latifolia, Citrus x paradisi, Agave spp., Juglans regia L., Punica granatum L.; from the seeds of Pimpinella anisum, Elettaria cardamomum, Morinda citrifolia, Anethum graveolens, Foeniculum vulgare, Cuminum cyminum, Salvia officinalis, Salvia hispanica, Capsicum annauum, Rosa rubiginosa, Vitis vinifera, Cocos nucifera, Argemone mexicana L., Avena sativa from the fruits of Carum carvi, Coriandrum sativum, Laurus nobilis, Myristica fragans, Petroselinum crispum, Piper nigrum, Morinda citrifolia; on the stems or branches of Cinnamomum verum, Cedrus sp., Pinus sp., Eucalyptus sp., Abies sp., Cupressus sp., Agave spp., Aloe barbadenesis Mill.

[0097] The term biocide in a broad sense shall be understood as a chemical substance or of natural origin intended to destroy, counteract, neutralize, prevent action or exert any other type of control over any organism considered as harmful to both the animal and plant kingdoms. The foregoing includes viricides, antifungals, bactericides, pesticides, disinfectants, sanitizers, against all types of unicellular microorganisms, and germicide, etc.

[0098] Nevertheless, the term biocide could also be understood according to the context described, as established in Annex V of the Regulation on biocides of the EUROPEAN CHEMICAL AGENCY (ECHEA).

[0099] In the present application, the term nanomaterial will correspond to the definition adopted by the European Commission which establishes that a nanomaterial will be understood as a natural, incidental or manufactured material containing particles in an unbond state or as an aggregate or as an agglomerate and where, for 50% or more of the particles in the number size distribution, one or more external dimensions is in the size range 1 nm and 100 nm.

[0100] Germicide shall be understood as an agent that destroys germs that are harmful to both the animal kingdom and the plant kingdom.

[0101] The term nanocomposite refers to a one-dimensional, two-dimensional, three-dimensional system and amorphous materials, obtained from one or two components or materials on a nanometric scale, unless it is interpreted otherwise according to the context in which it is mentioned.

[0102] Transition metals will be understood as those classified in the Periodic Table that are not considered as toxic metals to mammals and plants, for example, manganese, iron, cobalt, nickel, copper, zinc, silver, platinum, gold.

[0103] Toxic metals are those whose concentration in the environment can cause damage to both the animal and plant kingdoms.

[0104] Synthesis of Materials

[0105] TiO.sub.2ZnO nanocomposite was prepared by the sol-gel process using titanium (IV) butoxide (Aldrich 97%) and their respective salts as precursors: In a 3-neck flask, 30 to 50 mL of titanium (IV) butoxide, 4 g of polyethylene glycol (PEG) and 40 to 60 mL of ethanol (Aldrich 99.4%) were mixed. The obtained solution was heated to 60 to 120 C. under reflux and 18 mL of distilled water containing the appropriate average of Zn(NO.sub.3).Math.6H.sub.2O were added in a separation funnel in the preparation of the materials with 1.0, 3.0, 5.0 and 10.0% by weight, then a few drops of HNO.sub.3 (1 mol) were added to the solution of the respective metal salt until a pH=3 in the solution was achieved. Finally, the solution was added dropwise to abutoxide-ethanol solution (with a molar ratio of 8:1 butanol/water) after this the solution was mixed under magnetic stirring to form the gel. Then the solution was brought to 0 C. The solution was taken to the maturation process at 4 C.

[0106] The gel was then dried at 90 to 120 C. and the solid was ground to a fine powder in an agate mortar. The xerogel obtained was calcined between 400 and 600 C. in an air atmosphere, with a heating ramp of about 2 C./min, finally the product was ground again. As a reference, the pure TiO.sub.2 sample was prepared in the same manner but no salt was added.

[0107] Functionalization

[0108] In order to functionalize the mixed oxide, the material was placed in a test tube in a microwave oven and the extract was added in a range of between 10 ml to 50 ml, in a pressure reactor and at a temperature between 60 to 90 C. for 10 min, after completion of said time, the supernatant was removed and the sample was dried and taken to the ultrasonicator for one hour and then to a centrifuge for 10 minutes at 6000 rpm where the supernatant was removed and the NPs acquired a color from orange to red. The material was dried for 1 day at 60 to 90 C. to prevent volatilization of the desired bioactive compounds.

[0109] Obtaining the Extract

[0110] Material such as fruit peels and seeds were dried and ground and sieved with a 500 mesh. The dry matter was subjected to an ethanolic extraction using ultrasound-assisted extraction (UAE) (sonication). A dispersion with a 1:7.5 ratio of sample to solvent (95% EtOH) was prepared to a volume of 150 ml. The sonication conditions were between 40 and 60% amplifications, 0.6 s cycle for 10 to 30 min. After the UAE, the extracts were vacuum filtered using 20 membranes. The extracts were then concentrated in a rotary evaporator. The solvent residues were removed in a hood under air recirculation at room temperature and with magnetic stirring. Finally, the extracts were stored at room temperature for later use in formulating the emulsions.

[0111] Determination of Surface Areas and Size Distribution of Solid Pores

[0112] Nitrogen Physisorption

[0113] The specific areas of the samples were calcined at 500 C., which are reported in Table 1. The results showed that the specific area calculated by the BET method of the TiO.sub.2ZnO semiconductors was larger than that obtained with TiO.sub.2. A decrease in the areas is observed from 159 to 85 m.sup.2/g as the Zn.sup.2+ content increases for the TiO.sub.2ZnO solids from 1 to 10%. This can be observed in the adsorption-desorption isotherms (FIG. 1) since the ones with the largest volume is the TiO.sub.2ZnO material at 1% by weight of Zn. The tendency of the materials can also be observed as the Zn content increases.

[0114] The isotherms of the materials (a-b) in FIG. 1, including pure TiO.sub.2, were identified as type IV, typical features of mesoporous materials. In addition, the samples exhibited three different types of hysteresis: type H1 for 3%, type H2 for pure TiO.sub.2, (1%) and 5%, and type H3 for the treatments (10%) and (TiO.sub.2). The hysteresis loop associated with isotherms is attributed to capillary condensation of N.sub.2 gas that occurs in the pores, which also confirms the presence of a mesoporous structure. The change in the hysteresis loop could be due to the existence of smaller or larger pores in the samples, when metal was synthesized with TiO.sub.2 (Cu, Co, Ni, Cr, Pd, Zn and Sn) support.

TABLE-US-00001 TABLE 1 Nitrogen Physisorption ZnO Pore (% by Area Diameter weight) (m.sup.2/g) (nm) 1.0 159 7.8 3.0 104 9.4 5.0 102 9.6 10.0 85 7.6 TiO.sub.2 64 6.5

[0115] The crystalline structure of pure TiO.sub.2 and mixed oxides was determined by X-ray diffraction (XRD) analysis (see FIG. 2). Diffractograms of the materials show the anatase phase of TiO.sub.2 corresponding to peaks at 2=25.3, 37.9, 47.8, 54.5, 55, 62.5, 69, 70, 75 and 82, with a respective Miller index of (101), (103), (200), (105), (211), (204), (116), (220), (215) and (303) planes (JCPDS 21-1272). Furthermore, the characteristic diffraction peaks of the ZnO structures around 2=31.7, 34.5, 36.3, 47.5, 56 and 62.7, are not observed, therefore these results suggest that some Zn.sup.2+ cations were incorporated into the titanium dioxide (or titania) lattice, as evidenced by the increase in the cell parameter with respect to the Zn content (Table 1). However, due to the high specific area shown in mixed oxides, it is highly likely that ZnO is so dispersed forming conglomerates on the surface of titanium dioxide (or titania) and is not detectable by XRD.

[0116] RAMAN Spectroscopy

[0117] The Raman spectra of the TiO.sub.2ZnO samples are shown in FIG. 3. All Raman peaks are characteristic of the anatase phase. Bands at 145 cm.sup.1, 395 cm.sup.1, 513 cm.sup.1 and 640 cm.sup.1 are related to the TiO.sub.2 nanocrystals, which were previously identified. The intensity of the peaks decreases as Zn.sup.2+ content increases, indicating a significant decrease in the crystallinity of the mixed oxides. These results are consistent with those obtained by XRD, where the smallest crystal size corresponds to the highest Zn.sup.2+ content.

[0118] Diffuse Reflectance UV-Vis Spectroscopy

[0119] UV-Vis spectra were performed to investigate the effect of ZnO on the photophysical properties of TiO.sub.2ZnO semiconductors. FIG. 4 shows the diffuse reflectance UV-Vis spectra of the materials. All samples show an optical absorption below 400 nm, which can be attributed to the electronic TiO transition of the TiO.sub.2 nanocrystals. The results show a small shift in the red region (3.12-3.03 eV) for TiO.sub.2ZnO samples compared to the reference of anatase phase of the TiO.sub.2 semiconductor (3.2 eV). Thus, the effect of ZnO within the TiO.sub.2 lattice exerts only small variations in the forbidden energy band.

[0120] For the evaluations of the TiO.sub.2ZnO functionalized extract, seven reference strains (6 bacteria and 1 yeast) according to the American Type Culture Collection (ATCC), and two control bacterial strains (see Table 2) were used:

TABLE-US-00002 TABLE 2 Strains used in the extract evaluations ID number 1.-Aeromonas hydrophyla ATCC 12600 (reference strain) 2.-Escherichia coli ATCC 8739 (reference strain) 3.-Enterococcus fecalis ATCC 10541 (reference strain) 4.-Staphylococcus aureus ATCC 12600 (reference strain) 5.-Candida albicans ATCC 90028 (reference strain) 6.-Listeria monocytogenes ATCC 19115 (reference strain) 7.-Escherichia coli ATCC 25922 (control strain) 8.-Staphylococcus aureus ATCC 25923 (control strain) 9.-Salmonella enterica subsp. ATCC 9150 (reference strain) enterica serovar Paratyphi

[0121] All the nine strains were inoculated in three different media at a concentration of 0.5 according to the McFarland nephelometer. No growth of any of the strains was observed in both concentrations 9:1 and 9.5:5 after incubating 48 h at 37 C. In contrast, growth of all strains was indeed observed in GN (culture medium without extract) (FIG. 1).

[0122] FIGS. 5a to 5d show the distribution and the manner in which the TiO.sub.2ZnO nanoparticles agglomerate (a): 1.0% Zn, (b): 3.0% Zn, (c): 5.0% Zn and (d): 10.0% Zn).

[0123] FIGS. 6a-6d, corresponding to a transmission electron microscopy (TEM) study, show in detail the morphology, size (20 nm) and crystallization of the nanosystem obtained by the method of the present invention.

[0124] FIGS. 7a-7c show the Evaluation of 7 ATCC reference strains and 2 control strains: 7a) nutritive agar medium without extract; 7b) nutritive agar with 100% plant extract, 7c) nutritive agar with 50% plant extract. On each plate: A1-D1 Aeromonas hydrophyla ATCC 12600, A2-D2 Escherichia coli ATCC 8739, A3-D3 Enterococcus fecalis ATCC 10541, A4-D4 Staphylococcus aureus ATCC 12600, A5-D5 Candida albicans ATCC 90028, A6-D6 Listeria monocytogenes ATCC 19115, E1-F2 Escherichia coli ATCC 25922 (control strain), E3-F4 Staphylococcus aureus ATCC 25923 (control strain), E5-F6 Salmonella enterica subsp. enterica serovar Paratyphi AATCC 9150.

[0125] Therefore, samples 9:1 and 9.5:5 do inhibit the growth of Aeromonas hydrophyla ATCC12600, Escherichia coli ATCC 8739, Enterococcus fecalis ATCC 10541, Staphylococcus aureus ATCC 12600, Candida albicans ATCC 90028 and Listeria monocytogenes ATCC 19115 and Salmonella enterica subsp. enterica serovar Paratyphi AATCC 9150 strains.

[0126] FIG. 8 is a photomicrograph showing the variations in size and nanospheric or nano-droplet shape of the nanoparticles of the nanosystem, for which the particle sizes shown therein are not entirely representative of the real size, since it was only an attempt to obtain the size of the particles of the nanosystem, in addition to the fact that one skilled in the art should understand that the determination of particle size is more reliable through the TEM characterization (as in FIGS. 5a to 6d) than in the SEM study.

Examples and Results

[0127]

TABLE-US-00003 BIOLOGICAL EFFECTIVITY (BE) TESTS IN AGRICULTURAL CROPS ACCORDING WITH THE Official Mexican Standard NOM-032-FITO-1995 COMMON NAME COMMON NAME SCIENTIFIC NAME SCIENTIFIC NAME OF THE DISEASE OF THE CROP OF THE CROP OF THE PATHOGEN AND/OR PATHOGEN Fungi (fungi spp.) Tomato Solanum Phytophthora Late blight Aubergine Lycopersicum infestans Chili Solanum Pepper melongena Tobacco Capsicum Potato annum Capsicum annum Nicotiana tabacum Solanum tuberosum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare, commercial, control cooper sulphate 350 g per hectare, and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 83% of control and caused no negative phytotoxic effects on the crop. Coffee Coffea Hemilela Coffe leaf rust vastatrix Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare, commercial, control cooper sulphate 350 g per hectare, and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 85% of control and caused no negative phytotoxic effects on the crop. Potato Solanum Fusarium sp. Dry rot tuberosum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare, commercial, control cooper sulphate 350 g per hectare, and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosytem dose was 1.0 liter per hectare with an 81% of control and caused no negative phytotoxic effects on the crop. Papaya Carica papaya Colletotrichum Anthracnose sp Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. EB measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 81% of control and caused no negative phytotoxic effects on the crop. Raspberry Rubus idaeus Botrytis sp. Botrytis fruit rot Strawberry Fragaria Blackberry mexicana Blueberry Rubus Currant ulmifolius Vaccinium myrtillus Ribes rubrum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare, commercial, control cooper sulphate 350 g per hectare, and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 84% of control and caused no negative phytotoxic effects on the crop. Avocado Persea Phytophthora Avocado americana spp. sadness Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 84% of control and caused no negative phytotoxic effects on the crop. Bell Pepper Capsicum Pythium Damping off annuum aphanidermatum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L of the active agent water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 86% of control and caused no negative phytotoxic effects on the crop. Bell Pepper Capsicum Rhizoctonia Damping off annuum solani Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L of the active agent water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 87% of control and caused no negative phytotoxic effects on the crop. Cut Rose Rosa canina Peronospora sp. Peronospora Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 89% of control and caused no negative phytotoxic effects on the crop. Cut Rose Rosa canina Podosphaera Powdery pannosa mildew Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Cut Rose Rosa canina Verticillum spp. Yellowing Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 89% of control and caused no negative phytotoxic effects on the crop. Green tomato Physalis Phytophthora Late blight ixocarpa infestans Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 89% of control and caused no negative phytotoxic effects on the crop. Banana Musa Sigatoka negra Sigatoka paradisiaca Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 86% of control and caused no negative phytotoxic effects on the crop. Pumpkin Cucurbita pepo Sphaerotheca Powdery fuliginea mildew Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 87% of control and caused no negative phytotoxic effects on the crop. Cucumber Cucumis Sphaerotheca Powdery mildew sativus fuliginea Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test Statistic design: complete random blocks with five method treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Cocoa Theobroma Moniliophthora Witches' broom cacao perniciosao Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 90% of control and caused no negative phytotoxic effects on the crop. Cocoa Theobroma Crinipellis Witches' broom cacao perniciosao Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 85% of control and caused no negative phytotoxic effects on the crop. Mango Mangifera Fusarium Witches' broom indica moniliforme Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 86% of control and caused no negative phytotoxic effects on the crop. Mango Mangifera Fusarium Witches' broom indica oxysporum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 86% of control and caused no negative phytotoxic effects on the crop. Avocado Persea Colletotrichum Anthracnose americana acuatatum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Test method Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Avocado Persea Sphaceloma Avocado scab americana perseae Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 85% of control and caused no negative phytotoxic effects on the crop. Garlic Allium sativum Sclerotium White rot cepivorum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Onion Allium cepa Pyrenochaeta Pink root terrestris Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 89% of control and caused no negative phytotoxic effects on the crop. Onion Allium cepa Sclerotium White rot cepivorum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Barley Hordeum Puccinia hordei Barley rust vulgare Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: of the nanosystem that is being treatments, three complete random blocks with five tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 90% of control and caused no negative phytotoxic effects on the crop. Bean Phaseolus Uromyces Bean rust vulgaris phaseoli Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 84% of control and caused no negative phytotoxic effects on the crop. Peach Prunus persica Sphaerotheca Powdery panosa mildew Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 82% of control and caused no negative phytotoxic effects on the crop. Strawberry Fragaria Botrytis cinerea Botrytis fruit rot mexicana Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 86% of control and caused no negative phytotoxic effects on the crop. Olive Olea europaea Cycloconium Peacock eye oleaginea Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Carrot Daucus carota Rhizoctonia Crater rot carotae Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: of the nanosystem that is being treatments, three complete random blocks with five tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 81% of control and caused no negative phytotoxic effects on the crop. Grape Vitis vinifera Botrytis cinerea Gray-mold rot Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Raspberry Rubus idaeus Pucciniastrum Rust americanum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Guava Psidium Colletotrichum Guava cloves guajava gloeosporioides Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 87% of control and caused no negative phytotoxic effects on the crop. Mexican Lemon Citrus x Phytophthora Gummosis Persian Lime aurantiifolia citrophthora Orange Citrus Grapefruit limettioides Citrus x sinensis Citrus aurantium Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 90% of control and caused no negative phytotoxic effects on the crop. Mexican Lemon Citrus x Fusarium Root rot Persian Lime aurantiifolia oxysporum Orange Citrus Grapefruit limettioides Citrus x sinensis Citrus aurantium Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayere calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 91% of control and caused no negative phytotoxic effects on the crop. Tomato Solanum Phytophtora Smut and wilt in Lycopersicum capsici chili and pepper Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO fungicidal activity (percentage of inhibition) of the nanosystem that is being tested with 1:2500, 1:5000, and 1:7500 ml of product/ml of water; absolute control. The culture medium was used as product diluent, with 4 repetitions for each treatment. All treatments were cultured at the same time and incubated at 28 C. for 6 and 8 days according to the growth of each microorganism. Measurements were made every 24 h. Results The optimal dose of nanosystem was 1 mL in 2500 mL with 100% of inhibition. Tomato Solanum Fusarium Wilt Lycopersicum oxysporum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO fungicidal activity (percentage of inhibition) of the nanosystem that is being tested with 1:2500, 1:5000, and 1:7500 ml of product/ml of water; absolute control. The culture medium was used as product diluent, with 4 repetitions for each treatment. All treatments were cultured at the same time and incubated at 28 C. for 6 and 8 days according to the growth of each microorganism. Measurements were made every 24 h. Results The optimal dose of nanosystem was 1 mL in 2500 mL with 100% of inhibition. Tomato Solanum Fusarium Vascular wilt Lycopersicum Lycopersici Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO fungicidal activity (percentage of inhibition) of the nanosystem that is being tested with 1:2500, 1:5000, and 1:7500 mL of product/mL of water; absolute control. The culture medium was used as product diluent, with 4 repetitions for each treatment. All treatments were cultured at the same time and incubated at 28 C. for 6 and 8 days according to the growth of each microorganism. Measurements were made every 24 h. Results The optimal dose of nanosystem was 1 mL in 2500 ml with 100% of inhibition. Tomato Solanum Rhizoctonia Smut and root rot Lycopersicum solani Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO fungicidal activity (percentage of inhibition) of the nanosystem that is being tested with 1:2500, 1:5000, and 1:7500 mL of product/mL of water; absolute control. The culture medium was used as product diluent, with 4 repetitions for each treatment. All treatments were cultured at the same time and incubated at 28 C. for 6 and 8 days according to the growth of each microorganism. Measurements were made every 24 h. Results The optimal dose of nanosystem was 1 mL in 2500 ml with 100% of inhibition. Bacteria Prickly pear Opuntia spp. Erwinia spp. White rot (nopal) Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliageprickly pear (or flower) with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with an 88% of control and caused no negative phytotoxic effects on the crop. Lettuce Lactuca sativa Escherichia coli E. coli Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 98% of control and caused no negative phytotoxic effects on the crop. Blue Agave Agave tequilana Erwinia spp. Agave's bacteria Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 92% of control and caused no negative phytotoxic effects on the crop. Potato Solanum Clostridium spp. Botulism tuberosum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 93% of control and caused no negative phytotoxic effects on the crop. Tomato Solanum Streptomyces Common scab Lycopersicum scabies Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO; culture media Mueller- Hinton in complete blocks with four repetitions in each treatment, three of the nanosystem that is being tested with 1:1000, 1:2000, and 1:4000 mL of product/mL of water; absolute control. Incubated at 28 C. for 48 hours with measurements made every 24 h. Bactericidal activity parameter (inhibition halos) was measured in vitro by the Kirby-Bauer method. Results The optimal dose of nanosystem was 1 mL in 2500 mL with 100% of inhibition. Tomato Solanum Xantomona Bacterial spot Lycopersicum euvesicatoria Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO; culture media Mueller- Hinton in complete blocks with four repetitions in each treatment, three of the nanosystem that is being tested with 1:1000, 1:2000, and 1:4000 mL of product/mL of water; absolute control. Incubated at 28 C. for 48 hours with measurements made every 24 h. Bactericidal activity parameter (inhibition halos) was measured in vitro by the Kirby-Bauer method. Results The optimal dose of nanosystem was 1 mL in 1000 mL with control of 13.01 mm. Tomato Solanum Ralstonia Bacterial wilt Lycopersicum solanacearum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO; culture media Mueller- Hinton in complete blocks with four repetitions in each treatment, three of the nanosystem that is being tested with 1:1000, 1:2000, and 1:4000 mL of product/mL of water; absolute control. Incubated at 28 C. for 48 hours with measurements made every 24 h. Bactericidal activity parameter (inhibition halos) was measured in vitro by the Kirby-Bauer method. Results The optimal dose of nanosystem was 1 mL in 1000 mL with control of 14.05 mm. Tomato Solanum Pseudomona Tomato pith Lycopersicum corrugata necrosis Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistical design: IN VITRO; culture media Mueller- Hinton in complete blocks with four repetitions in each treatment, three of the nanosystem that is being tested with 1:1000, 1:2000, and 1:4000 mL of product/mL of water; absolute control. Incubated at 28 C. for 48 hours with measurements made every 24 h. Bactericidal activity parameter (inhibition halos) was measured in vitro by the Kirby-Bauer method. Results The optimal dose of nanosystem was 1 mL in 1000 mL with control of 12.87 mm. Tomato Solanum Clavibacter Bird's eye, Lycopersicum michiganensis bird's eye spot, and bacterial wilt Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Viruses Chili Capsicum PVH Huasteco virus annum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 92% of control and caused no negative phytotoxic effects on the crop. Serrano chili Capsicum TMV Tobacco pepper annuum mosaic Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 91% of control and caused no negative phytotoxic effects on the crop. Bell Pepper Capsicum TEV Tobacco Etch annuum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 94% of control and caused no negative phytotoxic effects on the crop. Habanero chili Capsicum CMV Cytomegalovirus annuum Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 94% of control and caused no negative phytotoxic effects on the crop. Pasilla Pepper Capsicum TMV Tobacco annuum mosaic Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 91% of control and caused no negative phytotoxic effects on the crop. Papaya Carica papaya PRSV Papaya Virus Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 92% of control and caused no negative phytotoxic effects on the crop. Melon Cucumis melo CYSDV Yellow Stunting Watermelon Citrullus lanatus Virus Zucchini Cucurbita pepo Cucumber Cucumis sativus Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 91% of control and caused no negative phytotoxic effects on the crop. Tomato Solanum TBSV Tomato stunt Lycopersicum viroid Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 90% of control and caused no negative phytotoxic effects on the crop. Chili Capsicum PHYVV Pepper yellow annuum vein Dose per 0.75, 1.0 and Concentration of 10% hectare 1.25 L in 200 L the active agent of water (nanosystem) Test method Statistic design: complete random blocks with five treatments, three of the nanosystem that is being tested with 0.75, 1.0 and 1.25 L per hectare; commercial control, cooper sulphate 350 g per hectare; and Absolute control with no application. Total applications: 5 at 7-day intervals. BE measurement parameters and phytotoxicity: Townsend and Heuberger Formula; Variance analysis, Turkey's multiple comparison test. Phenological state of the crop: Growing period Application was done on foliage with a motorized knapsack sprayer calibrated according to the doses used. Results The optimal nanosystem dose was 1.0 liter per hectare with a 91% of control and caused no negative phytotoxic effects on the crop.

TABLE-US-00004 BIOLOGICAL EFFECTIVITY (BE) TESTS IN ANIMAL SPECIES (LIVESTOCK, ZOOTECHNICS, VETERINARY) COMMON NAME OF THE SCIENTIFIC DISEASE COMMON SCIENTIFIC NAME OF THE AND/OR NAME NAME PATHOGEN PATHOGEN Hen Gallus gallus domesticus Aspergillus Aspergillosis fumigatu. Dose per animal 500 ml in 1000 L of water Concentration of 10% the active agent (nanosystem) Fungi (fungi spp.) Test method Statistical design: 3 groups of fowl were given the nanosystem as follows: 2 preventive treatments with dilutions 0.5 ml:1000 ml (TP-1) and 1 ml:1000 ml (TP-2), and 1 control group: Control 1 (C1) with no application of nanosystem; with conventional farm management. 100 fowls were included per group. An exclusive corral was assigned in which three 3 3 m-squares were prepared. Total applications: 2 (weekly). BE measurement parameters and cytotoxicity: Applications were made in the drinking water. Physical inspections of vomit were made on the animal's feed until this adverse effect was contained. Lessons in proventriculus and mortality were registered. Results The optimal nanosystem dose was 1.0 liter in 1000 L with a control of 95% and caused no negative cytotoxic effects in this species. Bacteria Bees Apis mellifera Melissococcus European plutonius foulbrood Dose per 5.0 and 10 mL in 1000 mL Concentration of 10% application the active agent (colony) (nanosystem) Test method Statistical design: 3 groups of which 2 groups of colonies were given the nanosystem as follows: 2 curative treatments with dilutions 5 ml:1000 ml (TP-1) and 10 mi:1000 ml (TP-2), and 1 colony as a control group: Control 1 (C1) with no application of nanosystem; with conventional management schedule of tetracycline complex. 4 colonies were included per group. Total applications: 2 (weekly). BE measurement parameters and cytotoxicity: Applications were made as fine spray (fog effect). Physical checks of dead organisms were made. Daily measurements were made for two weeks. Results Both of the doses were statistically equivalent with a control of 98% and caused no negative cytotoxic effects in this species. Cattle Bos Taurus/indicus Syndrome: Metritis Arcanobacterium pyogenes Fusobacterium n. Bacteroides spp. Escherichia coli Dose per 10 mL in 1000 mL Concentration of 10% application the active agent (intrauterine) (nanosystem) Test method Statistic design: Five groups as follows: 4 cow groups were given the nanosystem with curative treatment with dilutions 1 ml, 2.5 ml, 5 ml, 7.5 ml:1000 ml, 1 control group of cows with application of commercial antibiotics (Neomycin) and conventional managing schedule. 4 cows per group. Total applications: 3 (weekly) BE measurement parameters and cytotoxicity: Application was made by intrauterine infiltration with syringe. Physical inspection was made and swab samples were taken for ELISA test in petri dishes in lab. Daily measurements were made for two weeks. Results The optimal control dose was 10 mL. Diluted in 990 mL of distilled water with a 98% of control and caused no negative cytotoxic effects in this species. All cows treated excluding those of the control group, showed these results within 21 days after starting the applications. Bovines Bos Taurus/indicus Mycobacterium Tuberculosis Ovines Ovis aries tuberculosis Caprine Capra aegagrus hircus Dose per 20 mL not diluted Concentration of 10% application the active agent (intravenous, (nanosystem) jugular vein) Test method Statistic design: Five groups as follows: 4 cow groups were given the nanosystem with curative treatment with a single 20 ml dose, 1 control group of cows with application of commercial antibiotics (Streptomycin) and conventional managing schedule. 15 cows per group. Total applications: 2 (initial and after 20 days) BE measurement parameters and cytotoxicity: Application was made by intravenous injection in the jugular vein. Physical inspection was made and blood samples were taken before starting the application, and after 30 days for PCR laboratory tests. Positive diagnosis of M. tuberculosis in the first test and negative result after 30 days. Results The optimal control dose was 20 mL with a 92% of control and caused no negative cytotoxic effects in this species. Bovines Bos Taurus/indicus Brucella abortus Brucellosis Ovines Ovis aries Goats Capra aegagrus hircus Dose per 20 mL not diluted Concentration of 10% application the active agent (intravenous, (nanosystem) jugular vein) Test method Statistic design: Five groups as follows: 4 cow groups were given the nanosystem with curative treatment with a single 20 ml dose, 1 control group of cows with application of commercial antibiotics (Streptomycin) and conventional managing schedule. 15 cows per group. Total applications: 2 (initial and after 20 days) BE measurement parameters and cytotoxicity: Application was made by intravenous injection in the jugular vein. Physical inspection was made and blood samples were taken before starting the application, and after 30 days for PCR laboratory tests. Positive diagnosis of B. abortus in the first test and negative result after 30 days. Results The optimal control dose was 20 mL with a 92% of control and caused no negative cytotoxic effects in these species. Fowl Gallus gallus Escherichia coli Avian diarrhea domesticus Dose per 5.0 and 10 mL in 1000 mL Concentration of 10% application the active agent (nanosystem) Test method Statistical design: 3 groups of which 2 groups of fowl were given the nanosystem as follows: 2 curative treatments with dilutions 5 ml:1000 ml (TP-1) and 10 ml:1000 ml (TP-2), and 1 fowl group as a control group: Control 1 (C1) with no application of nanosystem; with conventional management schedule of Amoxycillin complex. 4 animals were included per group. Total applications: 2 (weekly). BE measurement parameters and cytotoxicity: Applications were made by oral route. Physical check of dead organisms was made. Daily measurements were made for two weeks. Results Both of the doses were statistically equivalent with a control of 93.5% and caused no negative cytotoxic effects in this species.

TABLE-US-00005 BIOLOGICAL EFFECTIVITY (BE) TESTS IN AQUATIC SPECIES COMMON NAME OF THE SCIENTIFIC DISEASE COMMON SCIENTIFIC NAME OF THE AND/OR NAME NAME PATHOGEN PATHOGEN Bacterial control Tilapia Oreochromis niloticus Pseudomonas sp. i Pseudomonasis Dose per 200 mL in 1000 L Concentration of 8% application the active agent (nanosystem) Test method Statistical design: 3 groups of which 2 groups of tilapia in bucket of water with a capacity of 1,000 L received the nanosystem, with 1 healing treatment with dilution 200 ml:1000 lt and 1 group of control tilapia (C1) without application of nanosystem, with the conventional management schedule. Each group with 500 organisms each. Total applications: 2 (15 days). BE measurement parameters and cytotoxicity: The administration was directly carried out in dissolution of the container with physical review of the effect of dead organisms. Daily measurements were made for two weeks. Results Both of the doses were statistically equivalent with a control of 89.5% and caused no negative cytotoxic effects in this species. Neon tetra Paracheridon Pleistophora Neon disease innesi hyphessobryconis Dose per 200 mL in 1000 L Concentration of 8% application the active agent (nanosystem) Test method Statistical design: 3 groups of which 2 groups of neon tetra in bucket of water with a capacity of 1,000 L received the nanosystem, with 1 healing treatment with dilution 200 ml:1000 lt and 1 group of control Neon tetra (C1) without application of nanosystem, with the conventional management schedule. Each group with 500 organisms each. Total applications: 2 (15 days). BE measurement parameters and cytotoxicity: The administration was directly carried out in dissolution of the container with physical review of the effect of dead organisms. Daily measurements were made for two weeks. Results Both of the doses were statistically equivalent with a control of 89.5% and caused no negative cytotoxic effects in this species. Virus White shrimp Panaeus vannamei White spot White Spot syndrome virus Syndrome (WSSV) Dose per 200 ml in 1000 L Concentration of 8% application the active agent (nanosystem) Test method Statistical design: 3 groups of which 2 groups of white shrimp in bucket of water with a capacity of 1,000 L received the nanosystem, with 1 healing treatment with dilution 200 ml:1000 L and 1 group of control white shrimp (C1) without application of nanosystem, with the conventional management schedule. Each group with 4000 organisms each. Total applications: 2 (15 days). BE measurement parameters and cytotoxicity: The administration was directly carried out in dissolution of the container with physical review of the effect of dead organisms. Daily measurements were made for two weeks. Results Both of the doses were statistically equivalent with a control of 83.5% and caused no negative cytotoxic effects in this species. Fowl Gallus gallus AIBV (avian Avian infectious domesticus coronavirus) bronchitis, Massachusetts strain Dose per 5.0 and 10 mL Concentration of 10% application in 1000 mL the active agent (nanosystem) Test method Statistical design: 3 groups of which 2 groups of fowl were given the nanosystem as follows: 2 treatments with dilutions 5 ml:1000 ml (TP-1) and 10 ml:1000 ml (TP-2), and 1 fowl group as a control group: Control 1 (C1) with no application of nanosystem; contact time, 15 minutes. Total applications: 1 during the test (7 days). BE measurement parameters and cytotoxicity: The application was made by fine spraying (fog effect). Physical check of dead organisms was made. Daily measurements were made for one week. Results Both of the doses were statistically equivalent; in treated groups no embryonic mortality or characteristic lesions were found. A 100% of viral INACTIVATION was obtained and caused no negative cytotoxic effects in this species. Bees Apis mellifera BQCV Black queen cell Dose per 5.0 and 10 mL Concentration of 10% application in 1000 mL the active agent (colony) (nanosystem) Test method Statistical design: 3 groups of which 2 groups of colonies were given the nanosystem as follows: 2 curative treatments with dilutions 5 ml:1000 ml (TP-1) and 10:1000 (TP-2), and 1 colony as a control group: Control 1 (C1) with no application of nanosystem; with conventional management schedule of tetracycline complex. 4 colonies were included per group. Total applications: 2 (weekly). BE measurement parameters and cytotoxicity: Applications were made as fine spraying (fog effect). Physical check of dead organisms was made. Daily measurements were made for two weeks. Results Both of the doses were statistically equivalent with a control of 99% and caused no negative cytotoxic effects in this species. Fowl Gallus gallus VENC Newcastle domesticus Disease virus Dose per 5.0 and 10 mL in Concentration of 10% application 1000 mL the active agent (nanosystem) Test method Statistical design: 3 groups of inoculum in chicken embryo: where 2 groups were given the nanosystem, as follows: 2 treatments with dilutions 5 ml:1000 ml (TP-1) and 10 ml:1000 ml (TP-2), and 1 fowl group as a control group: Control 1 (C1) with no application of nanosystem; contact time, 15 minutes. Total applications: 1 during the test (7 days). BE measurement parameters and cytotoxicity: The application was made by fine spraying (fog effect). Physical check of dead organisms was made. Daily measurements were made for one week. Results Both of the doses were statistically equivalent; in treated groups no embryonic mortality or characteristic lesions were found. A 100% of viral INACTIVATION was obtained and caused no negative cytotoxic effects in this species. Fowl Gallus gallus VIA Avian influenza domesticus Dose per 5.0 and 10 mL in Concentration of 10% application 1000 mL the active agent (nanosystem) Test method Statistical design: 3 groups of inoculum in chicken embryo: where 2 groups were given the nanosystem, as follows: 2 treatments with dilutions 5 ml:1000 ml (TP-1) and 10 ml:1000 ml (TP-2), and 1 fowl group as a control group: Control 1 (C1) with no application of nanosystem; contact time, 15 minutes. Total applications: 1 during the test (7 days). BE measurement parameters and cytotoxicity: The application was made by fine spraying (fog effect). Physical check of dead organisms was made. Daily measurements were made for one week. Results Both of the doses were statistically equivalent; in treated groups no embryonic mortality or characteristic lesions were found. A 100% of viral INACTIVATION was obtained and caused no negative cytotoxic effects in this species. Equines Equus caballus BPV Papilloma/ sarcoma Dose per 100 mL in 1000 mL Concentration of 10% application the active agent (nanosystem) Test method Statistical design: Cases: 2 horses received the nanosystem at a dilution of (topical dermal) 100 ml:1000 ml by topical application for 14 days. BE measurement parameters and cytotoxicity: The application was made by fine spraying (fog effect). Physical inspection of reduction of the tumor or critical mass. Daily measurements were made for up to one month after treatment starting. Results Evolution of total cicatrization. The tumor did not grow again as before when surgically extracted and after six months a new tumoral mass appeared. A 100% of viral INACTIVATION was obtained and caused no negative cytotoxic effects in this species. Canines Canis lupus VCPV-2 Parvovirus familiaris Dose per 100 mL in 1000 mL Concentration of 10% application the active agent (nanosystem) Test method Statistical design: Cases: 13 dogs (non vaccinated puppies not older than (oral route) 90 days) received the nanosystem. The first dose was undiluted (3 mL- syringe shot with no water). After that, the nanosystem was administered in the drinking water at a dilution of 10 ml:1000 ml for 5 days until recovery was evident. BE measurement parameters and cytotoxicity: The application was made by oral syringe shot and in drinking water. Physical inspection of reduction of symptoms. Daily measurements were made for up to 15 days after treatment starting. Results Evolution of symptoms with a 100% of viral INACTIVATION, and caused no negative cytotoxic effects in this species. In Vitro/In Vivo Sus scrofa PRRSV Porcine Swine domesticus reproductive and respiratory syndrome Dose per 0.005%. 0.01% Concentration of 10% application and 0.015% the active agent (nanosystem) Test method Statistical design: Lab-tek chambers were inoculated with a lung cell line which was infected according to the protocol. Three factors were considered to carry out the test: 1. Active agent concentration 2. Virus concentration 3. Incubation time First, the virus/active agent mixture was made at the three different concentrations of each, virus and activ eagent: 0.005%, 0.01% and 0.015% of active with a virus concentration of 106 DICT 50%/mL; these same active concentrations with virus concentrations of 105, and 104. Exposition time: 10 and 15 minutes. Immediately after this time, cells were inoculated and incubated at 37 C. for 1 h; then, the inoculum was removed and maintaining culture medium added. Each test was made in triplicate. Plates were fixed at 24, 48, and 72 hours for a Direct Immunofluorescence Test. Results The nanosystem was able to inactivate PRRSV at the indicated concentrations in an exposition time of 15 minutes at the maximal test concentration studied of one million (1 10.sup.6) viral particles per milliliter, causing a 100% of viral INACTIVATION and caused no negative cytotoxic effects in this species. In Vitro/In Vivo Sus scrofa H3N2 Porcine Swine domesticus H1N1 influenza Dose per 1:2500, 1:5000, Concentration of 10% application and 1:7500 mL the active agent (nanosystem) Test method Statistical design: Challenge with two virus strains: Influenza Type A H3N2 and H1N1 at a concentration of 1:256 HAU/50 L and then inoculated in MDCK cells to evaluate viral replication in the presence of the active agent that is being tested. Three dilutions were made: 1:2500 mL, 1:5000 mL, and 1:7500 mL. The nanosystem-virus mixture was left in contact during 15 minutes so that the effect on viruses could take place; then, 6-well plates with MDCK cells were inoculated with the different concentrations of nanosystem-virus, three serial passages were carried out, and after each passage a hemagglutination test was made. After the third passage, cells were harvested and a PCR test was carried out to confirm the diagnosis, Results The nanosystem was able to inactivate the two strains of porcine influenza virus type A, H3N2 and H1N1 at the concentrations 1:5000 mL, and 1:7500 mL after the third passage, causing a 100% of viral INACTIVATION and no negative cytotoxic effects.

[0128] From the above described, unless stated otherwise, that all numbers expressing ingredient quantities, reaction conditions, etc., used in this application and claims should be understood that constitute approximate quantities; therefore, they may vary depending on the desirable properties wanted to be obtained with the instant invention.

[0129] Furthermore, it is clear that other embodiments would be evident to a skilled person from the specification and practice of the invention described herein. Therefore, it is expected that the specification and examples herein are considered as illustrative only.

BIBLIOGRAPHY

[0130] 1. Katz, L.; Baltz, R. H. Natural product discovery: Past, present, and future. J. Ind. Microblol. Biotechno 2016, 43, 155-176. [0131] 2. Das, S.; Singh, V. K.; Dwivedy, A. K., Chaudhari, A. K.; Upadhyay, N.; Singh, P.; Sharma, S.; Dubey, N. K. Encapsulation in chitosan-based nanomatrix as an efficient green technology to boost the antimicrobial, antioxidant and in situ efficacy of Coriandrum sativum essential oil. Int. J. Biol. Macromol. 2019, 133, 294-305. [0132] 3. Shetta, A.; Kegere, J.; Mamdouh, W. Comparative study of encapsulated peppermint and green tea essential oils in chitosan nanoparticles: Encapsulation, thermal stability, in-vitro release, antioxidant and antibacterial activities. Int. J. Biol. Macromol. 2019, 126, 731-742, [0133] 4. Feyzioglu, G. C.; Tornuk, F. Development of chitosan nanoparticles loaded with summer savory (Saturejahortensis L.) essential oil for antimicrobial and antioxidant delivery applications, LWT 2016, 70, 104-110. [0134] 5. Ntohogian, S.; Gavriliadou, V.; Christodoulou, E.; Nanaki, S.; Lykidou, S.; Naidis, P.; Mischopoulou, L.; Barmpalexis, P.; Nikolaidis, N.; Bikiaris, D. N. Chitosan nanoparticles with encapsulated natural and uf-purified annatto and saffron for the preparation of uv protective cosmetic emulsions. Molecules 2018, 23, 2107. [0135] 6. Hussein, A. M.; Kamil, M. M.; Lotfy, S. N.; Mahmoud, K. F.; Mehaya, F. M.; Mohammad, A. A. Influence of nano-encapsulation on chemical composition, antioxidant activity and thermal stability of rosemary essential oil. Am. J. Food Technol. 2017, 12, 170-177. [0136] 7. Nedovic, V.; Kalusevic, A.; Manojlovic, V.; Levic, S.; Bugarski, B. An overview of encapsulation technologies for food applications. Procedia Food Sci. 2011, 1, 1806-1815. [0137] 8. Munin, A.; Edwards-Lvy, F. Encapsulation of Natural Polyphenolic Compounds; a Review. Pharmaceutics 2011, 3, 793-829. [0138] 9. Casanova, F.; Santos, L. Encapsulation of cosmetic active ingredients for topical applicationA review. J. Microencapsul. 2016, 33, 1-17. [0139] 10. Jafari, S. M. An overview of nanoencapsulation techniques and their classification. In Nanoencapsulation Technologies for the Food and Nutraceutical Industries; Academic Press: Cambridge, MA, USA; 2017; pp. 1-34. [0140] 11. Suganya, V.; Anuradha, V. Microencapsulation and Nanoencapsulation: A Review. Int. J. Pharm. Clin Res. 2017, 9. [0141] 12. Jyothi, N. V.; Prasanna, P. M.; Sakarkar, S. N.; Prabha, K. S.; Rarnaiah, P. S.; Srawan, G. Y. Microencapsulation techniques, factors influencing encapsulation efficiency. J. Microencapsul. 2010, 27, 187-197. [0142] 13. Gibbs, B. F.; Kermasha; S.; Inteaz, A.; Catherine, N.; Mulligan, B. Encapsulation in the food industry: A review. Int. J. Food Sci. Nutr. 1999, 50, 213-224. [0143] 14. Assadpour, E.; Jafari, S. M. Advances in Spray-Drying Encapsulation of Food Bioactive Ingredients: From Microcapsules to Nanocapsules, Annu. Rev. Food Sci. Technol. 2019, 10, 103-131. [0144] 15. Lohith Kumar, D. H.; Sarkar, P. Encapsulation of bioactive compounds using nanoemulsions. Environ. Chem. Lett. 2018, 16, 59-70. [0145] 16, Bakry, A. M.; Abbas, S.; Ali, B.; Majeed, H.; Abouelwafa, M. Y.; Mousa, A.; Liang, L. Microencapsulation of Oils: A Comprehensive Review of Benefits, Techniques, and Applications. Compr. Rev. Food Sol. Food Saf. 2016, 15, 143-182. [0146] 17. Coates, A. R.; Halls, G.; Hu, Y. Novel classes of antibiotics or more of the same. Br. J. Pharmacol. 2011 163, 184-194. [0147] 18. Bassetti, M.; Merelli, M.; Temperoni, C.; Astilean, A. New antibiotics for bad bugs: Where are we. Ann. Clin. Microbial. Antimicrob. 2013, 12, 22. [0148] 19. Leucuta, S. E. Nanotechnology for delivery of drugs and biomedical applications. Curr. Clin. Pharmacol. 2010, 5, 257-280. [0149] 20. Mahapatro, A.; Singh, D. K. Biodegradable nanoparticles are excellent vehicle for site directed in-vivo delivery of drugs and vaccines. J. Nanobiotechnol. 2011, 9, 55. [0150] 21. Gunn, J.; Zhang, M. Polyblend nanofibers for biomedical applications: Perspectives and challenges. Trends Biotechnol. 2010, 28, 189-197. [0151] 22. Vasita, R.; Katti, D. S. Nanofibers and their applications in tissue engineering. Int. J. Nanomed. 2006, 1, 15-30. [0152] 23. Guo, G.; Fu, S.; Zhou, L.; Liang, H.; Fan, M.; Luo, F.; Qian, Z.; Wei, Y. Preparation of curcumin loaded poly(epsilon-caprolactone)-polyethylene glycol)-poly(epsilon-caprolactone) nanofibers and their in vitro antitumor activity against glioma 91 cells. Nanoscale. 2011, 3, 3825-3832. [0153] 24. Yoo, J. J.; Kim, C.; Chung, C. W.; Jeong, Y. I.; Kang, D. H. 5-aminolevulinic acid-incorporated poly(vinyl alcohol) nanofiber-coated metal stent for application in photodynamic therapy. Int. J. Nanomed. 2012, 7, 1997-2005 [0154] 25. J. Moma, J. Baloyi, Modified titanium dioxide for photocatalytic applications, Photocatalysts-Applications and Attributes, Intech Open, 2018. [0155] 26. M. Landmann, E. W. G. S. Rauls, W. G. Schmidt, The electronic structure and optical response of rutile, anatase and brookite TiO.sub.2, J. Phys.: Condens. Matter 24 (19) (2012) 195503. [0156] 27. V. Etacheri, C. Di, J. Valentin, D. Bahnemann Schneider, C. S. Pillai, Visible-light activation of TiO.sub.2 photocatalysts: advances in theory and experiments, J. Photochem. Photobiol. C 25 (2015) 1-29. [0157] 28. W. Xie, R. Li et, O. Xu., Enhanced photocatalytic activity of Si-doped TiO.sub.2 under visible light irradiation, Sci. Rep. 8 (1) (2018) 1-10. [0158] 29. C. Dette, M. A. Perez-Osorio, C. S. Kley, P. Punke, C. E. Patrick, P. Jacobson, K. Kern, TiO.sub.2 anatase with a bandgap in the visible region, Nano Lett. 14 (11) (2014) 6533-6538. [0159] 30. M. E. Khan, M. M. Khan, B. K. B. K. Min, M. H. Cho, Microbial fuel cell assisted band gap narrowed TiO.sub.2 for visible-light-induced photocatalytic activities and power generation, Sci. Rep. 8 (1) (2018) 1-12. [0160] 31. A. Fujishima, T. X. Zheng, A. D. Tryk, Surf. Sci. Rep. J. 63 (2008) 515. [0161] 32. Y. Umemura, E. Shinohara, A. Koura, T. Nishioka, T. Sasaki, Photocatalytic decomposition of an alkylammonium cation in a Langmuir-Blodgett film of a titania nanosheet, Langmuir 22 (8) (2006) 3870-3877. [0162] 33. J. Zheng, H. Yu, X. Li, S. Zhang, Enhanced photocatalytic activity of TiO.sub.2 nanostructured thin film with a silver hierarchical configuration, Appl. Surf. Sci. 254 (6) (2008) 1630-1635. [0163] 34. T. Irugnanam, Effect of polymers (PEG and PVP) on the sol-gel synthesis of microsized zinc oxide, J. Nanomater. 2013 (2013) 7, Article ID 362175. [0164] 35. A. A. Haidry, J. Puskelova, T. Plecenik, P. During, J. Gregus, M. Truchly, A. Plecenik., Characterization and hydrogen gas sensing properties of TiO.sub.2 thin films prepared by sol-gel method, Appl. Surf. Sci. 259 (2012) 270-275. [0165] 36. M. Kashif, U. Hashim, M. E. Ali, K. L. Foo, S. M. U. Ali, Morphological, structural, and electrical characterization of sol-gel-synthesized ZnO nanorods, J. Nanomater. 2013 (2013) 7, Article ID 478942. [0166] 37. P. Sun, H. Liu, H. Yang, Synthesis and characterization of TiO.sub.2 thin films coated on a metal substrate, Appl. Surf. Sci. 256 (10) (2010) 3170-3173. [0167] 38. W. Guo, C. Xu, G. Zhu, C. Pan, C. Lin, Z. L. Wang, Optical fiber/TiO.sub.2-nanowirearrays hybrid structures with tubular counter electrode for dye-sensitized solar cell, Nano Energy 1 (1) (2012) 176-182. [0168] 39. R. Palomino-Merino, O. Portillo-Moreno, L. A. Chaltel-Lima, R. Gutierrez Perez, M. de Icaza-Herrera, V. M. Castano, Chemical bath deposition of PbS: Hg.sup.2+ nanocrystalline thin films, J. Nanomater. 2013 (2013) 6. [0169] 40. Z. S. Khalifa, H. Lin, S. Ismat Shah, Structural and electrochromic properties of TiO.sub.2 thin films prepared by metallorganic chemical vapor deposition, Solid Films 518 (19) (2010) 5457-5462. [0170] 41. C. Jiang, M. Y. Leung, W. L. Koh, Y. Li, Influences of deposition and postannealing temperatures on properties of TiO.sub.2 blocking layer prepared by spray pyrolysis for solid-state dye-sensitized solar cells, Solid Films 519 (22) (2011) 7850-7854. [0171] 42. B. Barrocas, O. C. Monteiro, M. E. Melo Jorge, Photocatalytic activity and reusability study of nanocrystalline TiO.sub.2 films prepared by sputtering technique, Appl. Surf. Sci. 264 (2013) 111-116. [0172] 43. M. Maaza, 35 Natural Dyes for Photonics Applications, 2014 [0173] 44. P. Yuvasree, K. Nithya, N. Neelakandeswari, Biosynthesis of silver nanoparticles from Aloe vera plant extract and its antimicrobial activity against multidrug-resistant pathogens, in: International Conference on Advanced Nanomaterials & Emerging Engineering Technologies, IEEE, 2013, pp. 84-86. [0174] 45. Makarov, W.; Love, A J.; Sinitsyna, O V; Makarova, S S.; Yaminsky, I V.; Taliansky, M E, Kalinina, N O. Green nanotechnologies: synthesis of metal nanoparticles using plants. Acta. Naturae, 6(1): 35-44. (2014). 306 J. M. Abisharani et al./Materials Today: Proceedings 14 (2019) 302-307 [0175] 46. Hoffmann M R, Martin S T, Choi W Y, Bahnemann D W. Environmental applications of semiconductor photocatalysis. Chem Rev; 95: 69-96. 1995. [0176] 47. Fujishima A, Rao T N, Truk D A. Titanium dioxide photocatalysis. J Photochem Photobiol C: Photochern, 1: 1-21. 2000. [0177] 48. Aadarsh Mishra, Analysis Of Titanium Dioxide And Its Application In Industry, Int. J. Mech. Eng. & Rob. Res., 3(3), 7 pages, 2014. [0178] 49. Gelis C, Girard S, Mavon A, Delverdier M, Pailous N, Vicendo P. Assessment of the skin photo protective capacities of an organomineral broad spectrum sunblock on two ex vivo skin models. Photodermatol Photoimmunol Photomed; 19: 242-253. 2003. [0179] 50. Trouiller B, Reliene R, Westbrook A, Solaimani P, Schiestl R H. Titanium dioxide nanoparticles induce DNA damage and genetic instability in vivo in mice. Cancer Res; 69: 8784-8789. 2009. [0180] 51. Carlos Martin Shiva Ramayoni. Estudio de la actividad antimicrobiana de extractos naturales y cidos orgnicos. Posible alternativa a los antibiticos promotores de crecimiento. Department de Sanitat i d'Anatomia Animals. Facultat de Veterinarria. Universitat Autnoma de Barcelona