Introduction

Land snails are among the agricultural pests that have attracted widespread attention in recent years as a result of the damage they can cause to crops in general1. Many fruit and vegetable trees, as well as ornamental plants, have been identified as hosts for this pest2. The continual race between researchers in numerous sectors of pest control offers many answers to the process of population diminution involving synthetic pesticides, pesticides of plant origin, natural substances, and others with a nanostructure3,4. In recent years, numerous plant components, such as flowers, roots, stems, and fruit, are used in the environmentally friendly synthesis of metal-oxide and ferrite NPs as fuel for combustion5,6,7,8,9,10. The green synthesis is an economical and sustainable method that can serve as a substitute for dangerous chemicals11. These factors are what spurred us to use plants as well as their extracts in the formation of nanomaterials. Engineered nanoparticles in general are used to perform a wide spectrum of toxicological studies against both aquatic12 and terrestrial gastropods13,14. Metal-based nanoparticles have earned a progressed attention for their numerous application; biomedical application, physicochemical properties and their safe property through the unexpected harm to human15,16,17. Many parameters were evaluated using a variety of snail species, such as exposure time, nanoparticle type, oxidative stress, and many other parameters18. Many studies have been conducted to evaluate the impact of NPs on aquatic and terrestrial gastropod animals13,19,20,21. Experiments are ongoing and the search is feverish and currently being conducted to find materials from various sources that can control these animal pests of terrestrial gastropods such as Monacha, the extent of whose harm is expanding day after day22. TiO2 was used as a toxic material for two different genera of snails (Danio rerio and Carassius gibelio); oxidative stress, genotoxicity, and many other parameters were measured as bio-response. The study results revealed a variability of alterations, including ROS production, lysosomal membrane breakdown, and damage to protein components such as DNA, lipid peroxidation, and protein carboxylation19. Cu-Pb-ferrite/TiO2 NPs was applied against the land snail Eobania vericulata; the open field control experiment was done with a satisfactory diminution percentage of 66.36 compared to the recommended molluscicide of 70.23%. Treating animals with the LC50 values; a biochemical and histological assessment was performed; a rupture for digestive cells and the foot’s epithelial layer was observed; furthermore, a significant raise of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), as well as a noticeable decrease in total protein (TP)23. Silver nanoparticles were applied to control gastropods in fresh water (Lymnaea stagnalis), and a substantial bioaccumulation rate in soft tissues demonstrated after 21 days of exposure, Also a high ROS production rate24. bismuth NPs are an effective applied toxicant for controlling the previous mentioned aquatic mollusk genus, Lymnaea luteola, with many variables such as oxidative stress, genotoxicity, and apoptosis dependent on exposure time and the concentration applied25. The toxic effect of TiO2 as a potential toxic nanomaterial was investigated against the snail Helix aspersa as a bioindicator using 70 and 140 µg/L concentrations for two successive weeks, by spraying method. The LC50 value was calculated at 544 µg/L, many biochemical parameters were assessed such as damage DNA, phagocytic activity and hematocyte count in addition to catalase (CAT) and superoxide dismutase (SOD)26. Within this research, a Ba–Ce–CuFO/TiO2 nanocomposite was prepared greenly using Eichhornia Plant aqueous extract and the structure was elucidated via different analytical methods. The important practical step of evaluating such materials in an open field experiment for controlling land gastropods took place with a satisfied reduction percentage, in addition to the positive influence of our target nanocomposite against other investigated parameters (biochemical variables and histology) that could support the control process.

Experimental

Materials

Titanium (IV) isopropoxide (Ti[OCH(CH3)2]4) ≥ 97.0%, Fe(NO3)3. 9H2O, Cu(NO3)2. 3H2O, Ba(NO3)2, Ce(NO3)2. 6H2O and NaCl (98% purity) were got from Sigma Aldrich company. Citric acid, ammonia, ethanol, acetone, NaOH and H2SO4 were got from Merck company. All materials were used without any purification.

Instrumentation

The Perkin Elmer fourier transform infrared spectroscopy (FTIR) (2000 System-USA) was utilized to characterize the samples. The X-ray diffraction (XRD) D2 Bruker Phaser, Bruker, Germany, equipped with a 2θ = 10–70° and Cu Kα radiation, was employed for analyzing the crystal structure. Other tools included a JSM-6460LV field emission scanning electron microscopy (FE-SEM) coupled with an energy dispersive x-ray spectroscopy (EDS) detector, a vibrating sample magnetometer (VSM, Meghnatis Kavir Kashan Co., Kashan, Iran), and selected area electron diffraction (SAED), lattice fringe patterns, and a Philips CM30 were used to record high-resolution transmission electron microscopy (HR-TEM) pictures. ER 4102 common rectangular cavity was used at room temperature, the X-band EMX spectrometer (Bruker, Germany) acquired electron paramagnetic resonance (EPR) spectra. Perkin-Elmer RBD modified the PHI-5300 C ESC system with a monochromatic Mg-Ka excitation to determine X-ray photoelectron spectroscopy (XPS) spectra.

Ba–Ce–CuFO/TiO2 green preparation

Green TiO2 was synthesized using Eichhornia plant aqueous extract in agreement with our earlier research findings27. Green Ba–Ce–CuFO magnetic nanoparticles was prepared using a citrate sol–gel auto combustion method at room temperature28,29,30 (Fig. 1). The citric acid and total metal nitrates of ratio1:1, were dissolved in 50 ml of Eichhornia plant aqueous extract as prepared previously in our work27 with subsequent addition of citric acid31. Ammonia was then added until the pH of the solution reached 9. The solution was kept under magnetic stirring for 1 h and 75 °C Because of heating, water evaporates and viscous liquid forms which automatically combusts to give dark brown fluffy powder, then 0.3 g of green TiO2 prepared previously was added under vigorous magnetic stirring for 1 h, the solutions are mixed under vigorous magnetic stirring and simultaneously heating at 75 °C for 4 h. After the solution was filtered and dried in oven at 100 °C. The mixture was repeatedly washed with ethanol, acetone, and distilled water before drying. This powder was ground and calcined at 800 °C for 3 h, with the heating rate of 5 °C/min in the open air to obtain a dense nano-powder of green Ba–Ce–CuFO/TiO2.

Fig. 1
figure 1

Schematic steps for green Ba–Ce–CuFO/ TiO2 nanocomposite preparation.

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Molluscicidal toxicity, application techniques, and LC50 values determination

Snails’ collection and adaptation

M. cartusiana snails were collected manually during the winter season from an infested farm in Mansoura University, juvenile snails were excluded, and only the healthy adult snails were kept for experimentation. Animals were transferred in cloth bags to the lab and kept in a moist, loamy soil glass box (50 × 30 × 20 cm). Gastropods were feeding lettuce (Lacuta sativa) leaves as a direct food source for two successive weeks for lab adaptation32; the lab conditions were 25 ± 5 °C and 75 ± 5% humidity.

Leaf dipping

Fresh lettuce green leaves were engrossed in each treatment for 1 min. The leaves were put in plastic containers with sterilized, humid, sandy soil. Each box contained ten adult M. cartusiana snails. Muslin material was used to cover each box and held with rubber bands to keep the snails from exiting. Four concentrations using distilled water (500, 1000, 1500 and 2000 ppm) were prepared. The treatments were replicated three times. Mortality rates were determined after one, three, five, and seven days of treatment2 for the LC50 value determination.

Contact

It is identified as the contact method or thin layer approach, and it has been authorized as a toxicity evaluation method. Four different aqueous concentrations using distilled water (250, 500, 750 and 1000 ppm) of Ba–Ce–CuFO/ TiO2 were prepared. The examined materials were put on the surface of a Petri dish (9 cm in diameter) in three duplicates, each with ten snails. Two mL of each nanomaterial were dispersed all over the inner surface of the Petri dish by gently moving in circles. The snails were exposed to all treatments, and mortality rates were calculated after one, three, five, and seven days23.

Biochemical analysis

As a physiological response, Ba–Ce–CuFO/ TiO2 LC50 effects of the on biochemical parameters were investigated. Gastropods were treated with LC50, and after 3 days of treatment, the visceral mass was extracted by carefully cutting around the shell whorl with a particular bone scissor.

The parameters assessed included AST, ALT33, ALP34, and TP35. Tissues were homogenized using a suitable buffer solution. The homogenates were centrifuged at 3000 rpm for 20 min at 5 °C in a freezing centrifuge. The residues were rejected, while the supernatants were kept for determination of biochemical activities. For the control group, identical experiments were conducted.

Histopathological assessment

Two sets of snails were prepared; one was treated with water and the other with Ba–Ce–CuFO/ TiO2 LC50 values. The shell was discarded by its removal through cutting around the whorl with a precise bone scissor, starting from the opening aperture and ending with the whorl carefully with continuous removing of fragmented pieces. A (Zeiss binuclear) dissection microscope was set up, and a 10% prepared formalin solution was used to preserve the obtained foot and digestive gland samples for 24 successive hours. Paraffin wax was utilized for immersion of the tissue after dehydration and clearance with ethanol and xylene, respectively. A microtome is used for the paraffin block section with 5 μm thickness units. Glass slides were used for collecting the tissues, eosin and hematoxylin were used for staining the tissues. The photographing process was taking place with a lab microscope camera (Novel model NLCD, Lamp S-LED W1) for the histopathological assessment36.

Open field poison bait application

The trial was applied to a widely infested clover farm with M. catusiana gastropods, at Dakahlia Governorate, Egypt. 2 g for both the Ba–Ce–CuFO/ TiO2 nanocomposite and a Neomyl 20% SL; recommended molluscicide, was prepared using 5 g of sugarcane and 93 g of bran in a humid bait formulation, completing the 100 g. Humid sugarcane and bran without any toxicants were used as controls. Each treatment was replicated thrice. The applied poisonous baits were offered in 100 g, each replicated over moisture colorless plastic pieces. A pretreatment count of live snails per square meter was taken before applying the experiment. Live snails in the treatment, toxicant nanocomposite, and controls were recorded after 1, 3, 7, and 14 days. The diminution percentage was considered according to the Henderson and Tilton Eq. 137:

$${\text{Reduction}}\% = 100\left[ {1 - \frac{{t_{2} r_{1} }}{{t_{1} r_{2} }}} \right]$$
(1)

rl and r2 are numbers of live gastropds in untraeted plots before and after application, t1 and t2 are numbers of live gastropds in treated plots before and after application, respectively.

Statistics analysis

Utilizing Probit analysis, the LC50 was calculated, in ppm units. One way ANOVA was applied to all calculations, which were all represented as mean ± SE. Using Tukey’s approach, a confidence with a 95% level were computed. A probability of 0.05 or less was regarded as significant. Utilizing Cohort Software, all statistical analysis was done38.

Results and discussion

Characterization

Figure 2a displays the XRD patterns of spinel cubic green Ba–Ce–CuFO/TiO2 nanocomposite, by comparing XRD of green Ba–Ce–CuFO/TiO2 to green TiO2 sample XRD pattern from the prior findings39, The XRD pattern verified the existence of Ba–Ce–Cu-ferrite in green Ba–Ce–CuFO/TiO2 composite. Using Scherer’s Eq. 2, the average size of the crystals of green Ba–Ce–CuFO/TiO2 nanocomposite were determined40,

$$d_{{{\text{XRD}}}} = {\text{ }}0.{\text{89}}\lambda {\text{/}}\beta {\text{cos}}\theta$$
(2)

Where λ is the X-ray wavelength and β is the half maximum of diffraction lines full width. Ba–Ce–CuFO/TiO2 greenly synthesized nanocomposite with crystal size calculated is 20 nm.

In Fig. 2b, the bands in the region up to 700 cm− 1 are assigned to the fundamental vibrations of the ferrite crystal lattice. the absorption band (υ1) at 613 cm− 1 was ascribed to the tensile vibration of the tetrahedral complex and the vibration of the band (υ2) and octahedral complex at 479 cm− 141,42. A close examination of the FTIR spectra in Fig. 2b reveals an absorption peak at 479 cm− 1, which is indicative of intrinsic stretching vibrations of the metallic substance at the tetrahedral position. On the other hand, the octahedral–metal stretching is responsible for the lowest band, v2−, which is detected in the 400 cm− 1 region and lower. It is common for Fe3+ ions to be found in tetrahedral and octahedral positions43. The bands observed at 3382 and 1616 cm− 1 attributed to H–O–H stretching and bending patterns were interpreted as evidence of free (or absorbed) water44. The stretching vibration of (M-O) corresponds to the strong peak at 1616.9 cm− 1 and 1119.6 cm− 1, confirming the production of the metal-oxygen in ferrite-based. When TiO2 suspension were added to Ba–Ce–CuFO/TiO2, the 1458.3 cm− 1 band of absorption, which corresponded to the nitrates that emerged when the Ce, Cu, and Fe functional groups and their interactions left vanished45,46. The spectra show a distinctive bands at 479 cm− 1 (metal ion– oxygen (Mocta–O) at octahedral-site) and 613 cm− 1 (metal ion– oxygen (Mtetra–O) at tetrahedral-site), that are the -Fe2O4 groups significant bands, as a result of the green Ba–Ce–CuFO/TiO2 nanocomposite fabrication47. The FT-IR measurement confirmed by XRD results, therefore, also confirmed the formation of spinel structures. In the present work, occupation of Ba2+ in octahedral B site with larger ionic radii is responsible for the observed shift in absorption bands compared to our previous work29.

Fig. 2
figure 2

XRD pattern (a), ATR-FTIR spectra (b) of greenly prepared Ba–Ce–CuFO/TiO2 nanocomposite.

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The surface morphology of green Ba–Ce–CuFO/TiO2 (Fig. 3a–b) has been studied by the usage of FE-SEM. It could be seen from Fig. 3a–b that sample have particles with spherical morphology. The existence of the holes and nano-porous features in the green Ba–Ce–CuFO/TiO2 surface could be ascribed to the successful formation during the calcination process at 800 °C compared to our previous work29. Figure 3c–d shows the HR-TEM images green Ba–Ce–CuFO/TiO2 after calcination at 800 °C, reveals a regular distribution of nano-cubic and nano-spheres with almost an average size of particle from 15 nm and up to 25 nm, which indicate the common nano-scale nature of the synthesized magnetic matrix. The particles have distributed uniformly and are spherical in shape. Their magnetic nature is the primary cause of the high aggregation shown in these TEM images. The XRD measurements and these outcomes accord well. Furthermore, the average distribution of particle sizes results was dependable with the values derived from the XRD spectra. EDS analysis was used to confirm the composition of the product. Typical EDS elemental analysis spectrum of the sample synthesized at 800 ∘C is shown in Fig. 4a. It was evident that the powders contained Fe, Ce, Cu, O, Ba and Ti elements without extra foreign signals which confirms the purity of our prepared nanocomposite.

Fig. 3
figure 3

FE-SEM images (a, b) and HR-TEM (c, d) spectra of greenly Ba–Ce–CuFO/TiO2 nanocomposite.

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The magnetic measurement of the green Ba–Ce–CuFO/TiO2 sample, obtained using a VSM instrument, is shown in Fig. 4b. Green Ba–Ce–CuFO/TiO2 ‘s S-shaped curve magnetization was displayed. At room temperature, this analysis was conducted. The saturation is 2.9862 emu/ g, respectively. Previous findings state that the first level of magnetization saturation for Fe3O4 is 50 emu/g. Fe3O4’s saturation magnetization dropped to 16 emu/g with TiO2 coating, and decreases to 2.38 for Cu-Ce ferrite/TiO2 nanocomposite, therefore these values confirm the achievement of the synthesis process of green Ba–Ce–CuFO/TiO2 nanocomposite29,48,49. The presence of non-magnetic TiO2 is responsible for the lower magnetic values50. Previous study of preparation of Fe3O4@TiO2 nanocomposite obtained a saturation magnetization intensity of 49.2 emu/g− 151. Nonetheless, the green Ba–Ce–CuFO/TiO2 nanocomposite that was synthesized still has superparamagnetic characteristics that are powerful enough for magnetic recovery. Using a standard permanent magnet, the green Ba–Ce–CuFO/TiO2 nanocomposite may be readily isolated from solid mixtures or aqueous solutions due to its superparamagnetic properties.

The BET specific surface area of green Ba–Ce–CuFO/TiO2 was 115 cm3g− 1 with a total pore volume of 0.28 cm3g− 1, which compares with TiO2 with BET-specific surface area of 68 cm3g− 1 and pore volume of 0.17 from earlier research52. The surface area of Fe3O4–TiO2 NPs was as much as 94.9 m2/g as obtained from the previous study53. Green Ba–Ce–CuFO/TiO2 nanocomposite has a comparatively large surface area when compared to pure TiO2 (Fig. 4c) owing to its nanosized particles, indicating that its surface area was enhanced by the modification process, which might be due to TiO2 growth being constrained in the presence of doped ferrite and its doped components. It is well known that substances with larger surface areas improve efficiency regarding surface reaction efficiency by offering more reaction sites. The pore volume of TiO2 increased following modification using the Ba–Ce–CuFO method.

Fig. 4
figure 4

EDS (a), Vibrating sample magnetic hysteresis loop (b), and N2 adsorption/desorption isotherms (c) of greenly prepared Ba–Ce–CuFO/TiO2 nanocomposite at room temperature.

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The high-resolution XPS spectrum of the Cu 2p core levels on the CPF/CT surface were displayed in Fig. 5a. Cu(I) and Cu(II) were ascribed to the two sub-peaks at 933.2 and 941.03.5 eV that were deconvoluted from the wide peak of Cu 2p3/254. The shake-up satellite structure, Fe 2p1/2, and Fe 2p3/2 were represented by the peaks in Fig. 5c at ∼725.7, ∼718.8, and ∼711.9 eV, respectively55. Two peaks with binding energies at around 711.9 and 714.9 eV were identified in the deconvoluted Fe 2p3/2 spectra, and they were ascribed to octahedral Fe(II) and tetrahedral Fe(III), correspondingly56. The XPS spectra of O 1s are shown in Fig. 5c. The three distinctive peaks at 530.2 eV and 531.4 eV were identified as surface lattice and adsorbed oxygen, respectively. Surface adsorbed oxygen was more fluid than surface lattice oxygen, making it more suitable for the catalyst’s catalysis. The large Ce 3d peaks of the green Ba–Ce–CuFO/TiO2 suggest that it contains multi-valence ions (Fig. 5d).

Five peaks with binding energies of 883.3, 890.04, 898.9, 908.5, and 917.4 eV can be used to fit the first peak of Ce 3d of the green Ba–Ce–CuFO/TiO2, suggesting the presence of Ce3+ and Ce4+ species57. Standard values and those documented in the literature for CeO2 and Ce2O3 are in good agreement with the peak forms and binding energies of Ce 3d58. The peaks at 782.6 eV and 793.01 eV in the green Ba-Ce–CuFO/TiO2 Nanocomposite are ascribed to Ba 3d5/2 and Ba 3d3/259, respectively, as shown in Fig. 5e. The high-resolution Ti 2p doublet of the TiO2 ( Fig. 5f) is visible in the green Ba–Ce–CuFO/TiO2 XPS data. The binding energies for the green Ba–Ce–CuFO/TiO2 nanocomposite are shown on the spectrum to be 459.1 eV for Ti 2p3/2 and 463.3 eV for Ti 2p1/2, respectively. These values are extremely near to those of the Ti4+ valence state, while the XPS spectra of Ti 2p TiO2’s spin-orbit splitting was seen in the literature60 XPS spectra, yielding the doublets “Ti 2p3/2” (binding energy 458.77 eV) and “Ti 2p1/2” (binding energy 462.19 eV), which correspond to Ti⁴⁺ in the TiO2 lattice. Furthermore, having the binding energy of 460.11 eV, the Ti3⁺ in Ti2O3 corresponds to the shoulder area of Ti 2p1/2. This shifting in the XPS peaks indicates the successful of carbon doping into TiO2 lattice61. The existence of Ba, Ce, Cu, Fe, Ti, and O is confirmed by the green Ba–Ce–CuFO/TiO2 XPS full scan spectrum. Fe 2p1/2 & 3/2 and Cu 2p1/2 & 3/2 XPS peak positions and band separations (Fig. 5a,b) matched the obtained XPS data of copper ferrite perfectly62. Additionally, the existence of a satellite peak verified that Cu and Fe were present in the Cu2+ and Fe2+ & 3+ oxidation states. Additionally, the Cu 2p1/2 & 3/2 peak intensity changed (Fig. 5a).

Fig. 5
figure 5

High-resolution XPS spectra of Cu 2p (a), Fe 2p (b), O 1s (c), Ce 3d (d), Ba 3d (e), and Ti 2p (e) of greenly prepared Ba–Ce–CuFO/TiO2 nanocomposite.

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Toxicity assessment test for greenly prepared Ba–Ce–CuFO/TiO 2 nanocomposite on M. cartusiana under laboratory conditions

Data in Table 1 Indicated the LC50 and LC90 values of green Ba–Ce–CuFO/TiO2 nanocomposite through leaf dipping method 1218.79 and 6886.23 ppm while, for contact were 289.19 and 870.48 ppm, respectively. Trials have been performed to investigate and identify more promising compounds that might be used as pesticides built on their source (synthetic, natural, or even nano compounds)63,64,65,66. Compounds with a low LC50 value have a noteworthy toxicity effect, whereas other compounds with high LC50 values do not have a noteworthy toxicity impact. Our nanocomposite displayed an undeniable molluscicidal significant impact via its LC50 using two well-established procedures: leaf dipping and contact32. Comparing our promising nanocomposite green Ba–Ce–CuFO/TiO2 with other compounds2 who reported a nano F-ZnO with 1381.55 and 237.51 ppm for LC50 values using the previous methods of application, leaf dipping and contact, respectively, while the LC50 values were 631.33 and 193.67 ppm for CuPb-ferrite/TiO223when the same previously mentioned methods were applied, respectively. Our composite LC50 values are significantly comparable to those of F-ZnO and CuPb-ferrite/TiO2; the LC50 is good likened to those reported composites controlling two types of gastropod mollusks, M. cartusiana and E. vermiculata.

Table 1 Toxicity assessment, LC50 and LC90 values of green Ba–Ce–CuFO/TiO2 against M. cartusiana under lab conditions.
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Biochemical parameters evaluation

Analyzing biochemical parameters (Table 2) reveals a significant rise in ALT and AST values when exposed to The LC50 value of green Ba–Cu–CeFO/TiO2 nanocomposite; 2500.7 and 3554.3 mU/mg protein, respectively, compared to 2191.0 and 3335.0 mU/mg protein of control values of. ALP values showed a decrease of 163.67 mU/mg protein from the control 198 mU/mg protein, also a considerable decrease in the TP of M. cartusiana gastropod when exposed to the LC50 value of green Ba–Cu–CeFO/TiO2 nanocomposite with a value of 11.76 mg/g, which is much lower percentage compared to the control, in which the percentage of total protein is 15.3 mg/g.

Table 2 Effect of green Ba–Cu-CeFO/TiO2 nanocomposite at different biochemical variables against M. Cartusiana gastropods.
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Histological examination

The digestive gland, also known as the hepatopancreas, its general description involves digestive tubules separated by connective tissue, many types of different functional cells are positioned in the tubules, like digestive cells, execratory cells, and calcium cells. The excretory cells number is much lower than that of the digestive ones26. Calcium cells are categorized as small cells with bulky nucleus (Fig. 6a).

Many histopathological changes were obviously recorded through the examined digestive gland when treated with our green Ba–Cu–CeFO/TiO2 nanocomposite, a noticeable width increase in the lumen of the most digestive tubules was also observed, as was a significant obliteration of the digestive cells with hemocyte infiltration accompanied by the appearance of pyknotic nuclei67(Fig. 6b).

Fig. 6
figure 6

M. cartusiana photomicrographic section in digestive gland (a): The control, showing the normal digestive cell containing lumen (L), execratory cells (ECs), execratory vacuoles (EVs), calcium cells (CCs), connective tissue (CT), and muscle fibre surrounding the digestive cells (M); (b): After treatment with the LC50 of Ba–Cu–CeFO/TiO2, showed different histological abnormalities like ruptured digestive cells (RDCs), destructed digestive tubules (DDT), an enlarged lumen (L), hemocyte infiltration (HI), and pyknotic nuclei (PN).

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These animals are commonly referred to as terrestrial gastropods. As a result, the foot is the abdomen, or the organ of movement in the animal, and the imbedded glands, as well as the connective tissue and the sole of the foot, are located there68 (Fig. 7a). Foot histological assessment (Fig. 7b–d) showed an appearance of deep folds accompanied by an empty space vacuole, with a noticeable epithelial cover layer rupture when treated with the LC50 of green Ba–Cu–CeFO/TiO2.

Fig. 7
figure 7

M. cartusiana photomicrographic section: (a) normal foot sole, connective tissue layer (CT), and embedded foot glands (EFG); (b) many empty clear spaces (ES) after exposure to LC50 of green Ba–Cu–CeFO/TiO2; (c) foot with deep folds (DF) and empty spaces (ES); (d) sole of the foot with ruptured epithelial cells (REC). Sections were stained with hematoxylin and eosin.

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Field application

Table 3 shows field application data for M. cartusiana gastropods exposed to green Ba–Cu–CeFO/TiO2 nanocomposite through the common bait’s method. The snail’s reduction percentages decreased after the first and third days of application to 51.25 and 71.91%, respectively. The residual effects (Mean reduction for 7–14 days exposure) of the NP-applied bait was 82.82%, and the average reduction was 72.2%, compared to Neomyl 20% SL as a recommended molluscicide, which gives a 74.27% reduction.

Table 3 Population diminution percentage for the applied nanocomposite Ba–Cu–CeFO/TiO2 and the recommended pesticide neomyl 20% SL in open field conditions.
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Discussion

The evaluated enzymes are mainly distributed in the cytoplasm and are released into the bloodstream after hepatocellular harm, resulting in an increase in serum levels, it was considered that the increase in ALT and AST after exposure to nanocomposite might be influenced by a series of factors including damage of muscle, toxic hepatitis, hepatopancreatic injury, and, intestine, which result in a noteworthy reduction in the activity level of ALP and TP concentrations in the hemolymph of B. alexandrina snails when subjected to ZnO NPs at sublethal doses69.

Decreased protein content and ALP values in gastropods are often corresponds to an increased levels of AST and ALT, especially when exposed to potentially toxic substances, including nanocomposites2. Eugenol (the main constituent in clove oil) was examined against different biochemical variables in the terrestrial snails Theba pisana, the animals exposed to eugenol at both sub-lethal effects had much higher γ-glutamyltransferase (γ-GT) and AST activities. Moreover, noticeably increase in ALT ratio using sub-lethal doses at all exposure period4. A significant increase for ALT, and AST values were clearly noticed, while the TP activity was decreased significantly when M. cartusiana snails after exposed to Methomyl (Copter 90%) pesticide after one, two, and three days of exposure using two applied concentrations 0.0.75 g/L and 0.180 g/L70.

In general, higher ALT and AST levels are related with lower TP even in mammals (the Norway rat, R. norvegicus) when exposed to synthetic organic compounds, where ALT and AST, as well as total protein, have been studied as biological response markers. When rats treated with two triazole acetamide derivatives were compared to controls, both AST and ALT levels increased significantly, but total protein levels decreased considerably71. NPs generally have a high surface area and a high dissolving potential, as well as the facility to enter the cellular membranes and cause cell damage15,72.

Field application assessment is an applicable real technique for testing accuracy, quality, and the capability for any potential toxic controlling materials including nanocomposites to be well characterized in open field conditions. The aspect of field application of any materials under investigations takes great importance over ordinary laboratory tests and assessments, especially in the field of pest control73. Many species of land snails were documented as destructive invasive pests: M. cartusiana, Eobania vermiculata, Succinea putris, Cochlicella acuta, Theba pisana, and Helicella vestalis. M. cartusiana had the highest number during the investigation period and was considered the main infestation everywhere. With the gradual population increasing from winter to reach the highest density in the spring season, the control process through field application tests is the true choice to eliminate these different invasive gastropods74. Bee venom and propolis have been applied in the field against M. cartusiana using the toxic bait technique, the experiment showed a considerable diminution using propolis (43.42%), while the venom was (20.09%) when compared to the recommended Agrinate that posed a (62.68) reduction percentage32. Field application of F-ZnO and ZnO NPs against the terrestrial gastropod (M. cartusiana) was tested in an open clover farm; the results obtained by applying these nanocomposites through the common bait method, at a concentration of 2%, showed a noticeable diminution in the total number of animals over the course of 21 days of taking the results, and the general mean of the reduction was 60.08 and 56.39% for F-ZnO and ZnO respectively, which is a satisfactory result compared to the recommended Neomyl molluscicide that poses 69.55% reduction percentage2. Field evaluation of another group of CuPb-ferrite/TiO2 nanocomposites against Eobania vermiculata (Müller) in an infested citrus farm also exhibited a noteworthy reduction mean of 66.36% of the pest population23. The field experiment results are like those of the prescribed molluscicide, which we can attribute to the presence of several elements combined in this nanocomposite. These elements can easily penetrate the animal’s organs and tissues alone or in combination, they can cause toxicity and cell damage, as evidenced by enzymes and histopathological examination of cells and tissues.

Conclusions

This study is one of many persistent attempts to search for and explore new green materials with biological activity within pest control operations, especially the land snail’s control. Ba–Cu–CeFO/TiO2 nanocomposite was effectively green manufactured by using of citrate sol–gel auto combustion method using aqueous extract of Eichhornia plant, and the sample was then heated for 3 h at 800 °C. The size of particle and crystallinity of the manufactured sample were confirmed using XRD, and the size of particle is about 20 nm. The existence of every metal ion and its associated electronic state was verified by XPS data. The presence of more than one metal in our green nanocomposite Ba–Cu–CeFO/TiO2 gives it a relative advantage in the ability to cause harm and reach more than one site to affect the animal, as the analysis showed many biochemical parameters were proved the effectiveness of our nanocomposite through the noticeable increase in AST and ALT levels and a clear decrease in TP and ALP values when treating gastropods with the LC50 value compared to control. The damage caused by our nanocomposite to the muscular foot and hepatopancreas was obvious in the histopathological assay. Field application of the nanocomposite proved the achievement of the control procedure, and the results were satisfactory: a 72.2% population diminution compared to the recommended local pesticide Neomyl 20% SL, which poses a 74.27% reduction. The relative safety represented by the field application through the bait technique by applying it in separated plastic pieces that don’t allow it to affect with the soil or irrigation water and also being easy to dispose of after the control process gave an additional relative advantage to our applied composite, in addition to the possibility of collecting the nanocomposite being tested using magnets, which is an easy and effective method through which we can recycle and use again in order to save costs and also preserve the environment from any possible source during control processes. Therefore, we strongly recommend using this green multiple metal nanocomposite in land snail control operations.