Prerpints.org logo
Preprint
Article

This version is not peer-reviewed.

Oviposition Deterrent Effect of a High Quality Natural Zeolite for the Olive Fruit Fly Bactrocera oleae, under Different Conditions of Temperature and Relative Humidity

A peer-reviewed version of this preprint was published in:
Insects 2024, 15(4), 256. https://doi.org/10.3390/insects15040256

Submitted:

12 February 2024

Posted:

13 February 2024

You are already at the latest version

Abstract
In recent years the number of available chemical pesticides has been significantly reduced and there have been an urgent need for discovery of alternative to chemical pesticides products, as among others the natural zeolites (zeolitic rocks). We determined the mineralogical and chemical composition of a specific and continuous layers of zeolitic rock sample (ZeotP) from Petrota, Ev-ros, Greece and evaluated its oviposition deterrent effect for the olive fruit fly Bactrocera oleae (Diptera: Terphritidae). The tested natural zeolite contained 70 wt. % Clinoptilolite, 18 wt. % Amorphous material, 7 wt. % Feldspars, 4 wt. % Cristobalite and 1 wt. % Quartz. We tested the oviposition deterrent effect of ZeotP mixed or not with an emulsifier adjuvant NU-FILM-P® in water and applied on the surface of olive fruits. The ZeotP oviposition deterrent effect for the ol-ive fly was very high under a series of tested temperatures (17°C, 20°C, 25°C and 30°C) and RHs (23%, 33%, 55%, 75%, 94%). In addition, the ZeotP residual deterrent effect after equable water spraying was high and similar to the respective effect of a pyrethroid insecticide (Decis®). Our results may contribute to an effective control of the olive fruit fly using an alternative to chemical pesticides natural zeolite (zeolitic rocks) product.
Keywords: 
olive fly
;  
control
;  
oviposition deterrent effect
;  
natural zeolite (zeolitic rock)
;  
clinoptilolite
;  
temperature
;  
relative humidity
Subject: 
Biology and Life Sciences  -   Insect Science

1. Introduction

The continuous efforts for a greener and more sustainable world, in order to protect the environment from industrial and agricultural uses, lead among others to the use of alternative to chemicals methods, efficient and friendly to the environment and public health. In agriculture, and specifically in pest control, the available chemical products (insecticides, herbicides, fungicides) have been significantly reduced in the last decades due to their multiple negative effects, such as soil and water contamination, toxicity to animal and humans and inefficient pest control due to the development of resistant populations [1,2]. Therefore, the development and use of alternative to chemical pesticides products such as the inert-sorbent materials (mineral- and dust-based products) could be a promising solution. In recent years, many studies have confirmed the effectiveness of inert dust such as kaolin, for the control of crop and stored products pests [2,3,4]. There are limited studies, however, concerning the insecticidal effect of zeolites and their use for pest control.
Zeolites are in abundance among microporous materials worldwide and characterized as hydrous, microporous crystalline aluminosilicate minerals of alkali and alkaline earth elements. They consist of a unique three-dimensional framework structure, with bonding silica (SiO4) and alumina (AlO4) tetrahedral units, connected together with the sharing of all the oxygen atoms. They can be also defined as crystalline inorganic polymers, with their frameworks forming pores and channels containing significant contents of water molecules, and exchangeable cations (calcium, potassium, sodium etc.) [1,2,3,4,5].
Approximately 67 types of zeolites occur naturally, and more than 200 types are synthetically prepared in the laboratory. Natural zeolites are hydrothermal and of mainly volcanic origin. They can occur both in crystallized forms found in igneous and metamorphic rocks, as well as in grains of smaller diameters accumulated in sedimentary rocks [6,7]. The main rock-type, which contains great amounts of zeolite minerals, is characterized as zeolitic rock. In nature, apart from the fibrous zeolites (erionite, mordenite, roggianite, mazzite, etc.), which are hazardous for animal and human health, other non-toxic types exist such as chabazite, phillipsite, clinoptilolite, heulandite, analcime. Among them, HEU-type zeolites (clinoptilolite – heulandite) are of great research and economic interest. They are characterized by the presence of tabular crystals (typically 1-100 µm in size), and the presence of micro/nano-pores, that create infinite three-dimensional frameworks of silica, alumina and oxygen atoms in a structure of 10- and 8-membered rings of dimensions (7.5×3.1 Å (V=23.25 Å3), 4.6×3.6 Å (V=16.56 Å3), and 4.7×2.8 Å (V=13.16 Å3) [7,8].
Due to their unique characteristics such as their porosity and ion-exchange properties, as well as their abundance, zeolites have generated worldwide interest for their use in a wide range of applications. Natural zeolites have been broadly used in industry, as catalysts, ion exchangers, absorbents for oil and spills, separation agents and agents for the removal of heavy metals from wastewaters [5,9,10,11,12,13]. Due to the International Agency for Research on Cancer (IARC) and Food and Drug Administration (FDA) classification of zeolites as, respectively, “non-toxic” and safe for human consumption, they are extensively used for agricultural and horticultural purposes, among others, as fertilizers, slow releasing carriers of fertilizers and insecticides, feed additives, in soil amendment, water purification and gas absorption [14,15].
The olive fruit fly, Bactrocera oleae (Diptera: Tephritidae), is the most harmful pest of olives worldwide causing serious damage to olive production [16]. It is a monophagous pest, as its larva feeds exclusively on olive fruit [17]. Adult females lay their eggs inside the olive fruit, and hatching larvae feed in the mesocarp causing fruit damage and olive oil quality deterioration [18,19], as well as premature fruit drop [20]. In Mediterranean regions, there may be more than five overlapping generations from late summer through the winter and early spring if fruits are present on the trees and field temperatures are suitable for development [21,22,23].
The olive fly's behavior is strongly dependent on its host fruit and is related to various physical and chemical plant stimuli. The olive fruit favors the ovarian maturation of the females due either to contact or volatile stimuli, as well as to bacteria that are present on its surface [21,24,25,26]. In addition, olive fruit, due to its volatile and contact chemical stimuli, favors mating success and egg production of the olive fly [23,27]. Certain olive fruit’s organic volatile compounds such as n-octane and α-pinene, as well as a mixture of n-octane, α-pinene, limonene, ethyl hexanol, nonanal, n-dodecane, decanal and n–tetradecane favored successful reproduction of the olive fruit fly [27].
The control of the olive fly is mainly based on the extensive use of chemical insecticides, which usually results in the development of olive fly populations with high levels of resistance and failure of effective control, and the presence of chemical residues in olives and olive oil [17]. It is, therefore, of crucial importance to develop innovative and environmentally safe methods and products for the control of the olive fly.
Previous studies have shown that inert dusts i.e., kaolin, diatomaceous earth (DE), and natural zeolites have an insecticidal efficacy with partially removing an insect’s outer cuticle (epicuticule) through abrasion by hard non-sorptive particles or by disrupting the epicuticle via absorption of epicuticular lipids to sorptive particles [28]. Both processes induce rapid water loss from the insect’s body and cause death by dehydration [29,30,31]. Natural zeolite was found to have a high insecticidal effect, when applied as dust, to wheat seeds infested with the rice weevil Sitophilus oryzae and the red flour beetle Tribolium castaneaum, and found high mortality percentages [31]. Zeolites are also toxic against other insect pests that infest stored products, such as Tribolium confusum, Oryzaephilus surinamensis the maize weevil Sitophilus zeamais and three other beetle pests in stored wheat [32,33,34].
In a recent preliminary study carried out by our research group, a high-quality Greek natural zeolite was found to have a high insecticidal effect for adults of the bean weevil Acanthoscelides obtectus [29]. This high-quality natural zeolite contained 92 wt. % HEU-type (clinoptilolite), 3 wt. % Mica + Clay minerals, 3 wt. % Quartz and 2 wt. % Feldspars, and has a high insecticidal effect under various temperature and humidity conditions [29].
The use of alternative to chemical insecticides products, such as inert dusts, with olive fly deterrent/repellent effects may contribute to overcome the negative consequences of the use of pesticides and efficiently protect olive production [17]. The use of oviposition repellent products, such as the earth inert dusts (kaolin, natural zeolites), appeared to be promising non-chemical methods for the control of insect pests. Kaolin is an inert dust which has been extensively used commercially with the trade name Surround® for the control of the olive fly [35]. Visual and chemical stimuli lead the female olive fly to oviposit into olive fruits [23,27,36,37,38]. Kaolin is commercially used against the olive fly, deterring egg laying, as the presence of the particles of the product on the fruit surface could be an obstacle for the olive fruit recognition by the females [39,40].
The aim of the present study is to characterize and test the insecticidal effect of a novel Greek natural zeolite product. We determined the particular mineralogical and chemical composition of a high quality Greek natural zeolitic rock, and evaluated its oviposition deterrent effect for the olive fruit fly B. oleae under different temperature and RH conditions.

2. Materials and Methods

2.1. Stock colony and experimental flies

Our laboratory colony of B. oleae was established with adults emerged from field-collected olive fruits in October 2021 from the area of Chalkidiki, northern Greece. The stock colony flies were maintained in wooden cages (30 x 30 x 30 cm) with wire screen sides in a climatic room at 25°C, relative humidity (RH) of 65% and a 16L:8D photoperiod. The experimental flies were second-generation descendants of the stock colony. They were reared from egg to pupae in olive fruits which were collected in early August and since then, have been maintained at 5 ± 1°C for 2–3 months before being offered to the stock flies for oviposition. For the experiments, stock females oviposited in olive fruit of the variety Megaritiki. After oviposition, the infested olives were placed in plastic cups, covered with wet hessian fabric, and kept in an incubator at a constant temperature of 20 °C, 70% relative humidity (RH) and 16L:8D photoperiod. The newly formed pupae were collected daily and placed in Petri dishes. Emerging females and males were maintained in wooden cages like those used for the stock colony at 25°C, 70% RH and 16L:8D photoperiod, and used in the experiments when they were ten days old. Stock and experimental flies were provided ad libitum with a protein liquid diet of water, yeast hydrolysate and sugar (v: w: w 5: 4: 1).

2.2. Natural zeolite characterization

The natural zeolite sample (ZeotP) used in our experiments was collected from specific and continuous layers of zeolite-rich volcaniclastic tuffs in the surrounding area of Petrota, Evros region, northern Greece. Petrographic investigation of ZeotP was performed on a thin section by polarized microscope. The mineralogical composition was determined by X-Ray diffraction method (XRD), with a Philips PW1710 diffractometer equipped with Ni-filtered CuKα radiation on randomly oriented sample, from 3° to 63° 2θ at a scanning speed of 1.2°/ min. Chemical composition of ZeotP was performed using the ICP-MS method. The cation exchange capacity (sorption ability) was measured by the ammonium acetate saturation (AMAS) method [41].

2.3. Oviposition deterrent test effect

A portion of ZeotP with the appropriate grain size distribution (<63 μm) was prepared for the experiments. ZeotP was pulverized in a mechanical agate mortar for approximately 10 minutes, and then sieved to a size <63 μm in a mechanical sieve shaker, for 15 minutes. The procedure was repeated until the entire amount of natural zeolite was sieved, and the processed dust was collected.
ZeotP dust suspended in water (5 g dust to 100 ml water) with or without the addition of the adjuvant NU-FILM-P® (1,5 %). For the experiments, olive fruits were immersed in ZeotP suspensions in glass jars for 20 seconds or as control in water. After immersion, the fruits were allowed to dry on the surface of a metal sieve, and then were transferred to the base of a plexiglass cage (20 X 20 X 20 cm). In each cage, 5 adult females of B. oleae and 5 olives were transferred. To evaluate the oviposition deterrent effect of the tested natural zeolites, we scored under a stereoscope (Zeiss Stemi 305®), the number of oviposition holes on the olive fruits after 3 and 6 days of olives’ exposure to the egg-laying females. In each treatment, there were four replicates (cages with olives and adult females). The experiments were carried out in a climatic room with a temperature of 25°C and a 16L:8D photoperiod.

2.4. Effect of temperature

The oviposition deterrent effect of ZeotP with the adjuvant NU-FILM-P® was evaluated at four constant temperatures using the same experimental procedure explained in the above paragraph. The plexiglass cages with the treated olive fruits and the experimental females were maintained in climatic incubators at a series of constant temperatures (17°, 20°, 25° and 30 °C) with a 16L:8D photoperiod. In each tested temperature, there were four replicates, i.e., plexiglass cages with 5 adult females and 5 olive fruits.

2.5. Effect of relative humidity (RH)

In this series of experiments, the oviposition deterrent effect of ZeotP was evaluated under 5 different relative humidity (RH) levels. For the experiments, the female flies with ZeotP treated olives were maintained at different RH levels (23, 33, 55, 75 and 94%), at 25 °C and a 16L:8D photoperiod. To maintain the flies at different RHs, we used sealed plastic containers (30 × 40 × 30 cm) with an appropriate saturated water salt solution at their bottom (5 cm deep) [42,43]. A metal net was placed approximately 3 cm above the surface of the salt solution and served as the floor for the cages with experimental flies and olives. These pot cages were modified cylindrical cups of 430 cm3 volume (6.5 × 9.5 × 10 cm, diameter top × diameter base × height) with two circular openings (diameter: 2 cm) covered with nylon mesh to allow adequate ventilation. The cups were placed upside down on a petri dish bearing five equidistant circular holes (diameter: 1.5 cm) where the treated olive fruits were placed. The pot cages with the B. oleae females and the treated olives were maintained for 3 and 6 days at the different RH levels and subsequently, the number of oviposition holes on the fruits was scored. The saturated salt solutions used in the experiments and the respective RHs inside the cages are shown in Table 1. In each RH level, there were four pot cages (replicates) housing five B. oleae females and five olive fruits.

2.6. Residual oviposition deterrent effect of ZeotP on olives after water spraying

We determined the residual oviposition deterrent effect of ZeotP after spraying the treated olives with water. For the experiments, olives were immersed in ZeotP, with and without the NU-FILM-P® adjuvant, as described in the aforementioned paragraph. After ~1 h of the ZeotP application, the treated olives were placed in glass petri dishes 9 cm in diameter in the basement a potter precision spraying tower (Buckard Manufacturing Co Ltd. Rickmansworth, Hertfordshire, UK) calibrated at 0,68 atm pressure, and sprayed with two constant quantities, 5 ml or 15 ml of water/min. As control, we used olives treated with the insecticide Decis® with or without NU-FILM-P® adjuvant. After water spraying, the olive fruits were transferred to plexiglass cages with B. oleae adult females, and the number of oviposition holes on the olives was scored after 3, 6 and 9 days. In each treatment, there were four replicates, i.e., plexiglass cages with 5 adult females and 5 olive fruits. The experiments were carried out at a climatic room with a temperature of 25°C and a 16L:8D photoperiod.

2.7. Statistical analysis

A one-way ANOVA was performed to detect whether temperature, relative humidity and water spraying affect the oviposition deterrent effect of the tested natural zeolite. Subsequent significant differences (P < 0.05) of means were further separated with Student–Newman–Keul’s test using the IBM SPSS Statistics 24.0 (IBM Corp., Armonk, NY, USA). Before data analysis, Kolmogorov–Smirnov’s and Levene’s tests were used to confirm normality and homogeneity of variances, respectively. In cases in which heterogeneity of variances was significant, appropriate data transformation (log transformation) was conducted before the analysis. In those cases where the transformations failed to satisfy the criteria for parametric analysis, the Kruskal–Wallis non-parametric ANOVA followed by the Mann-Whitney U test for all possible pairwise comparisons, was conducted.

3. Results

3.1. Natural zeolite characterization

Semi-quantitative estimates of the abundance of the mineral phases were derived from the XRD data using the intensity (counts) of certain reflections, as well as the density and mass absorption coefficient for CuKa radiation of the minerals present [11]. As shown in Table 2, ZeotP consists of 70 wt% HEU-type zeolite (clinoptilolite), 1 wt% Quartz, 4 wt% Cristobalite, 7 wt% Feldspars (K-feldspars and plagioclase), and 18 wt% amorphous material. The natural zeolite sample can be characterized as clinoptilolitic zeolitic rock. The cation exchange capacity (sorption ability) was measured 193 meq/100g.
Regarding the chemical composition (Table 3), ZeotP contains 68.43 wt.% SiO2 and 11.60 wt.% Al2O3. The content of CaO is 3.80 wt.%, K2O is 2.12 wt.%, whereas MgO content is 0.96 wt.%. Low amounts of Na2O, Fe2O3T and MnO were measured.
The mineralogical composition aligns both with the chemical composition and cation exchange capacity. Except SiO2 and Al2O3, ZeotP contains sufficient amounts of CaO and K2O (Table 3).

3.2. ZeotP oviposition deterrent effect

The application on olives of an aqueous solution of ZeotP with and without NU-FILM-P® resulted in a significant decrease in the number of eggs laid on them. As shown in Table 4, when the female flies had access to olive fruits for 3 and 6 days, a mean number of 107.0 and 168.5 eggs respectively were laid in the non-treated olives whereas no eggs were laid in olives treated with a mixture of ZeotP and NU-FILM-P®. An intermediate mean number of 16.2 and 59.0 eggs were laid in olives sprayed with ZeotP without NU-FILM-P®. These results show that the addition of NU-FILM-P® adjuvant significantly increased the oviposition deterrent effect of ZeotP.

3.3. Effect of temperature

As shown in Table 5, at all the tested temperatures, there was a significant oviposition deterrent effect of ZeotP (with or without the addition of NU-FILM-P®). The number of eggs laid in olives, after the application of ZeotP with or without NU-FILM-P®) was very low to zero, in all the tested temperatures. These results show that, irrespective of temperature conditions, the oviposition deterrent effect of ZeotP is high and in particular when applied in combination with the adjuvant NU-FILM-P®.

3.4. Effect of relative humidity (RH)

As shown in Table 6, in all RHs tested, there was a significant oviposition deterrent effect of ZeotP. The number of eggs laid in olives after the application of ZeotP was significantly lower compared to the control, at all levels of relative humidity (23, 33, 55, 75 and 94% RH).

3.5. Residual oviposition deterrent effect of ZeotP on olives after water spraying

As shown in Table 7, after spraying with water the ZeotP treated olives, the oviposition deterrent effect of ZeotP, with or without NU-FILM-P®, was remained high, and the number of eggs laid in the fruits was significantly lower than the respective number of eggs laid in the control (non-treated olives). However, in almost all the cases, the addition of NU-FILM-P® in ZeotP suspension, resulted in a higher residual oviposition deterrent effect after water spraying. The residual ZeotP oviposition deterrent effect after water spraying was similar to the respective effect of the pyrethroid insecticide Decis®.

4. Discussion and conclusions

Inert dusts such as kaolin, diatomaceous earth (DE) and natural zeolites have an insecticidal efficacy which is mainly due to disruption of the insect’s outer cuticle (epicuticule) through abrasion by hard non-sorptive particles or by adsorption of epicuticular lipids to sorptive particles [28]. Both processes induce rapid water loss from the insect’s body and cause death by dehydration [29]. The physical properties of natural zeolites such as particle size, shape, surface area etc., as well as the chemical and thermal stability that depends on the Si/Al ratios (i.e., the least stable natural zeolites have low Si/Al ratios) affect their activity [29]. Here, we used a high quality natural zeolite with unique characteristics such as high content of clinoptilolite and microporous minerals, high cation exchange capacity (sorption ability) and content of K and Ca.
Zeolites, are highly toxic against a number of insect pests that infest stored products, such as Sitophilus oryzae and Tribolium castaneaum [30], Tribolium confusum and Oryzaephilus surinamensis [31], Sitophilus zeamais [32,33] and bean weevils Acanthoscelides obtectus [29]. Among the earth’s inert dusts, kaolin has been extensively used commercially with the trade name Surround® for the control of the olive fly [34]. Our present results show that ZeotP, after appropriate processing, and application in the form of aqueous suspension on the surface of the olive fruit, can significantly prevent the egg laying of the olive fruit fly and therefore protect the olives’ injury. In addition, the adjuvant NU-FILM-P®, mixed with ZeotP significantly increases the oviposition deterrent effect. The XRD analysis of ZeotP has shown that quartz is in very low quantities (1 wt. %), and fibrous minerals are completely absent which makes ZeotP safe for human health.
Previous studies of our group have shown that a high-quality Greek zeolite has a high insecticidal activity for the bruchid Acanthoscelides obtectus, under different temperature and RH conditions [29]. Here, we found that ZeotP, mixed with the adjuvant NU-FILM-P® and applied on olives, has a high oviposition deterrent effect, under different temperature and RH conditions. This high oviposition deterrent effect of ZeotP with NU-FILM-P® is maintained even after spraying with water the ZeotP treated olives, and is similar to the respective effect of the pyrethroid insecticide Decis® (deltamethrine), which is extensively used in the field against the olive fly.
In conclusion, the studied high quality zeolitic rock, rich in HEU-type zeolite, has a high oviposition deterrent effect against the olive fly B. oleae, particularly in preventing adult females from laying their eggs on olive fruits. This prevention of oviposition may be due to the creation of a thin layer of natural zeolite on the surface of the olives which seems to mechanically prevent females to adhere to the surface of the fruit and lay their eggs. Adult females lay their eggs inside the olive fruit, and the hatching larvae feed in the mesocarp causing fruit damage and olive oil quality deterioration [18,19], as well as premature fruit drop [20]. The thin layer of ZeotP formed on the olive surface prevents the oviposition process and fruit injury at a range of different temperatures and RH levels. The combination of ZeotP with an adjuvant (NU-FILM-P®) increased the oviposition deterrent effect and is a promising approach for the olive fly control.
The use of alternatives to pesticides methods, such as natural zeolite products with olive fly oviposition deterrent effect, may contribute to overcome the negative consequences of the use of chemical pesticides and efficiently protect olive production [17]. Field experiments with ZeotP, according the requirements of European legislation for the specific applications, are required to confirm its oviposition deterrent effect and efficacy for the control of the olive fly.

Author Contributions

Conceptualization, D. Koveos (D.K.), N. Kantiranis (N.Ka.), N. Kouloussis (N.K.), and A. Filippidis (A.F.), S.K. Kovaiou (S.K.K.), and A. Kokkari (A.K.); Methodology, D.K, N.Ka., A.F., and N.K..; Software, S.K.K., A.K., and G.F.; Validation, D.K., N.Ka., N.K., A.F., S.K.K., and A.K.; Formal analysis, D.K., S.K.K., and A.K.; Investigation, D.K., N.Ka., N.K., A.F., S.K.K., A.K., and G.F.; Resources, D.K., N.Ka., and N.K..; Data curation, S.K.K., and A.K.; Writing—original draft preparation, S.K.K., and A.K.; Writing—review and editing, S.K.K., A.K., G.F., N.Ka., and D.K..; Visualization, S.K.K., and A.K.; Supervision, D.K., N.Ka., and N.K..; Project administration, D.K., and N.Ka. All authors have read and agreed to the published version of the manuscript.

Data Availability Statement

Not applicable.

Acknowledgments

The present study was conducted in interdisciplinary collaboration of the Laboratory of Mineralogy and Petrology, School of Geology, and the Laboratory of Applied Zoology and Parasitology, School of Agriculture, of the Aristotle University of Thessaloniki, Greece.

Conflicts of Interest

The authors declare no conflict of interest. All the authors approve its publication in your journal.

References

  1. Jha, B.; Singh, D.N. A review on synthesis, characterization and industrial applications of fly ash zeolites. J. Matter. Educ. 2011, 33, 65. [Google Scholar] [CrossRef]
  2. Novembre, D.; Gimeno, D. Synthesis and characterization of analcime (ANA) zeolite using a kaolinitic rock. Sci. Rep. 2021, 11, 1–9. [Google Scholar] [CrossRef]
  3. Christidis, G.E.; Moraetis, D.; Keheyan, E.; Akhalbedashvili, L.; Kekelidze, N.; Gevorkyan, R.; Sargsyan, H. Chemical and thermal modification of natural HEU-type zeolitic materials from Armenia, Georgia and Greece. Appl. Clay Sci. 2003, 24, 79–91. [Google Scholar] [CrossRef]
  4. Kantiranis, N.; Sikalidis, K.; Godelitsas, A.; Squires, C.; Papastergios, G.; Filippidis, A. Extra-framework cation release from heulandite-type rich tuffs on exchange with NH4+. J. Environ. Manage. 2011, 92, 1569–1576. [Google Scholar] [CrossRef]
  5. Filippidis, A.; Papastergios, G.; Kantiranis, N.; Filippidis, S. Neutralization of dyeing industry wastewater and sludge by fixation of pollutants in very high quality HEU-type zeolitic tuff. J Ecol Environ 2015, 2, 221–226. [Google Scholar]
  6. Gottardi, G.; Galli, E. Natural Zeolites, Mineral and Rocks. Springer: Berlin/Heidelberg, Germany, 1985, 18.
  7. Baerlocher, C.; McCusker, L.B.; Olson, D.H. Atlas of Zeolite Framework Types. Elsevier, 2001.
  8. Dyer, A.; Tangkawanit, S.; Rangsriwatananon, K. Exchange diffusion of Cu2+, Ni2+, Pb2+ and Zn2+ into analcime synthesized from perlite. Microporous Mesoporous Mater. 2004, 75, 273–279. [Google Scholar] [CrossRef]
  9. Król, M. Natural vs. synthetic zeolites. Crystals 2020, 10, 622. [Google Scholar] [CrossRef]
  10. Tsitsishvili, G.V.; Andronikashvili, T.G.; Kirov, G.N. Natural zeolites. Ellis Horwood Limited, 1992.
  11. Filippidis, A.; Kantiranis, N. Experimental neutralization of lake and stream waters from N. Greece using domestic HEU-type rich natural zeolitic material. Desalination 2007, 213, 47–55. [Google Scholar] [CrossRef]
  12. Filippidis, A.; Apostolidis, N.; Paragios, I.; Filippidis, S. Zeolites clean up. Indust. Miner. 2008, 487, 68–71. [Google Scholar]
  13. Filippidis, A.; Tziritis, E.; Kantiranis, N.; Tzamos, E.; Gamaletsos, P.; Papastergios, G.; Filippidis, S. Application of Hellenic Natural Zeolite in Thessaloniki industrial area wastewater treatment. Desalination Water Treat 2016, 57, 19702–19712. [Google Scholar] [CrossRef]
  14. Papastergios, G.; Kantiranis, N.; Filippidis, A.; Sikalidis, C.; Vogiatzis, D.; Tzamos, E. HEU-type zeolitic tuff in fixed bed columns as decontaminating agent for liquid phases. Desalination Water Treat 2017, 59, 94–98. [Google Scholar] [CrossRef]
  15. Cerri, G.; Farina, M.; Brundu, A.; Dakovi´c, A.; Giunchedi, P.; Gavini, E.; Rassu, G. Natural zeolites for pharmaceutical formulations: Preparation and evaluation of a clinoptilolite-based material. Microporous Mesoporous Mater. 2016, 223, 58–67. [Google Scholar] [CrossRef]
  16. Cataldo, E.; Salvi, L.; Paoli, F.; Fucile, M.; Masciandaro, G.; Manzi, D.; Masini, C.M.; Mattii, G.B. Application of Zeolites in Agriculture and Other Potential Uses: A Review. Agronomy 2021, 11, 1547. [Google Scholar] [CrossRef]
  17. Malheiro, R.; Casal, S.; Cunha, S.C.; Baptista, P.; Pereira, J.A. Olive volatiles from Portuguese Cultivars Cobrançosa, Madural and Verdeal Transmontana: Role in Oviposition Preference of Bactrocera oleae (Rossi) (Diptera: Τephritidae). PLoS One 2015, 10, 1–15. [Google Scholar] [CrossRef] [PubMed]
  18. Daane, K.M.; Johnson, M. Olive fruit fly: Managing an ancient pest in modern times. Annu. Rev. Entomol. 2010, 55, 151–169. [Google Scholar] [CrossRef] [PubMed]
  19. Gucci, R.; Caruso, G.; Canale, A.; Loni, A.; Raspi, A.; Urbani, S.; Taticchi, A.; Esposto, S.; Servili, M. Qualitative changes of olive oils obtained from fruits damaged by Bactrocera oleae (Rossi). HortScience 2012, 47, 301–307. [Google Scholar] [CrossRef]
  20. Tzanakakis, M.E. Insects and mites feeding on olive: Distribution, importance, habits, seasonal development and dormancy. Brill Academic Publishers, 2006.
  21. Tzanakakis, M.E.; Koveos, D.S. Inhibition of ovarian maturation in the olive fruit fly, Dacus oleae (Diptera: Tephritidae), under long photophase and an increase of temperature. Ann. Entomol. Soc. Am. 1986, 79, 15–18. [Google Scholar] [CrossRef]
  22. Koveos, D.S.; Tzanakakis, M.E. Effect of the presence of olive fruit on ovarian maturation in the olive fruit fly, Dacus oleae, under laboratory conditions. Entomol. Exp. Appl. 1990, 55, 161–168. [Google Scholar] [CrossRef]
  23. Koveos, D.S.; Tzanakakis, M.E. Diapause aversion in the adult olive fruit fly through effects of the host fruit, bacteria, and adult diet. Ann. Entomol. Soc. Am. 1993, 86, 668–673. [Google Scholar] [CrossRef]
  24. Neuenschwander, P.; Michelakis, G. Infestation of Dacus oleae (Gmel.) (Diptera: Tephritidae) at harvest time and its influence on yield and quality of olive oil in Crete. J. Appl. Entomol. 1978, 86, 420–433. [Google Scholar] [CrossRef]
  25. Fletcher, B.S.; Pappas, S.; Kapatos, E. Changes in the ovaries of olive flies (Dacus oleae (Gmelin)) during the summer, and their relationship to temperature, humidity and fruit availability. Ecol. Entomol. 1978, 3, 99–107. [Google Scholar] [CrossRef]
  26. Fletcher, B.S.; Kapatos, E. The influence of temperature, diet and olive fruits on maturation rates of female olive flies at different times of the year. Entomol. Exp. Appl. 1983, 33, 244–252. [Google Scholar] [CrossRef]
  27. Kokkari, A.I.; Pliakou, O.D.; Floros, G.D.; Kouloussis, N.A.; Koveos, D.S. Effect of fruit volatiles and light intensity on the reproduction of Bactrocera (Dacus) oleae. J. Appl. Entomol. 2017, 141, 841–847. [Google Scholar] [CrossRef]
  28. Kokkari, A.I.; Milonas, P.G.; Anastasaki, E.; Floros, G.D.; Kouloussis, N.A.; Koveos, D.S. Determination of volatile substances in olives and their effect on reproduction of the olive fruit fly. J. Appl. Entomol. 2021, 00, 1–15. [Google Scholar] [CrossRef]
  29. Floros, G.D.; Kokkari, A.I.; Kouloussis, N.A.; Kantiranis, N.; Damos, P. , Filippidis, A.; Koveos, D. S. Evaluation of the natural zeolite lethal effects on adults of the bean weevil under different temperatures and relative humidity regimes. J. Econ. Entomol. 2018, 111, 482–490. [Google Scholar] [CrossRef] [PubMed]
  30. Glenn, D.M.; Puterka, G.J.; Drake, S.R.; Unruh, T.R.; Knight, A.L.; Baherle, P. Particle film application influences apple leaf physiology, fruit yield, and fruit quality. J. Amer. Soc. Hortic. Sci 2001, 126, 175–181. [Google Scholar] [CrossRef]
  31. Andrić, G.G.; Marković, M.M.; Adamović, M.; Daković, A.; Golić, M.P.; Kljajić, P.J. Insecticidal potential of natural zeolite and diatomaceous earth formulations against rice weevil (Coleoptera: Curculionidae) and red flour beetle (Coleoptera: Tenebrionidae). J. Econ. Entomol. 2012, 105, 670–678. [Google Scholar] [CrossRef] [PubMed]
  32. Roumbos, C.I.; Sakka, M.; Berillis, P.; Athanassiou, C.G. Insecticidal potential of zeolite formulations against three stored-grain insects, particle size effect, adherence to kernels and influence on test weight of grains. J. Stored Prod. Res 2016, 68, 93–101. [Google Scholar] [CrossRef]
  33. Eroglu, N.; Sakka, M.K.; Emekci, M.; Athanassiou, C.G. Effects of zeolite formulations on the mortality and progeny production of Sitophilus oryzae and Oryzaephilus surinamensis at different temperature and relative humidity levels. J. Stored Prod. Res. 2019, 81, 40–45. [Google Scholar] [CrossRef]
  34. Kljajić, P.; Andrić, G.; Adamović, M.; Bodroža-Solarov, M.; Marković, M.; Perić, I. Laboratory assessment of insecticidal effectiveness of natural zeolite and diatomaceous earth formulations against three stored-product beetle pests. J. Stored Prod. Res 2010, 46, 1–6. [Google Scholar] [CrossRef]
  35. Saour, G.; Makee, H. A kaolin-based particle film for suppression of the olive fruit fly Bactrocera oleae Gmelin (Dip., Tephritidae) in olive groves. J. Appl. Entomol. 2004, 128, 28–31. [Google Scholar] [CrossRef]
  36. Katsoyannos, B.I.; Kouloussis, N.A. Captures of the olive fruit fly Bactrocera oleae on spheres of different colours. Entomol. Exp. Appl. 2001, 100, 165–172. [Google Scholar] [CrossRef]
  37. Rotundo, G.; Germinara, G.S.; De Cristofaro, A.; Rama, F. Identificazione di composti volatili in estratti da diverse cultivar di Olea europaea L. biologicamente attivi su Bactrocera oleae (Gmelin) (Diptera: Tephritidae). Boil. Lab. Ent. agr. Filippo Silvestri 2001, 57, 25–34. [Google Scholar]
  38. Solinas, M.; Rebora, M.; De Cristofaro, A.; Rotundo, G.; Girolami, V.; Mori, N.; Di Bernardo, A. Functional morphology of Bactrocera oleae (Gmel.) (Diptera: Tephritidae) tarsal chemosensilla involved in interactions with the host-plant. Entomologica 2001, 35, 103–123. [Google Scholar] [CrossRef]
  39. González-Núñez, M.; Pascual, S.; Cobo, A.; Seris, E.; Cobos, G.; Fernández, C.E.; Sánchez-Ramos, I. Copper and kaolin sprays as tools for controlling the olive fruit fly. Entomol. Gen. 2021, 97–110. [Google Scholar] [CrossRef]
  40. Caleca, V.; Rizzo, R. Tests on the effectiveness of kaolin and copper hydroxide in the control of Bactrocera oleae (Gmelin). IOBC WPRS 2007, 39, 111–117. [Google Scholar]
  41. Kantiranis, N.; Filippidis, A.; Georgakopoulos, A. Investigation of the uptake ability of fly ashes produced after lignite combustion. J. Environ. Manage. 2005, 76, 119–123. [Google Scholar] [CrossRef]
  42. Wexler, A.; Hasegawa, S. Relative Humidity-Temperature Relationships of Some Saturated Salt Solutions in the Temperature. J Res Natl Inst 1954, 53, 19–26. [Google Scholar] [CrossRef]
  43. Winston, P.W.; Bates, D.H. Saturated solutions for the control of humidity in biological research. Ecology 1960, 41, 232–237. [Google Scholar] [CrossRef]
Table 1. Saturated salt solutions used with the corresponding RHs and saturation deficit.
Table 1. Saturated salt solutions used with the corresponding RHs and saturation deficit.
Solutions Relative humidity (%) Saturation deficit (kPa)
LiCl.H2O 23 2.96
MgCl2.6H2O 33 2.25
Mg(NO3)2.6H2O 55 1.51
NaCl 75 0.84
KNO3 94 0.17
Table 2. Semi-quantative mineralogical composition (wt.%) and ion exchange capacity (meq/100g) of Petrota natural zeolite sample ZeotP.
Table 2. Semi-quantative mineralogical composition (wt.%) and ion exchange capacity (meq/100g) of Petrota natural zeolite sample ZeotP.
Sample ZeotP
HEU-type zeolite (clinoptilolite) 70
Quartz 1
Cristobalite 4
Feldspars 7
Amorphous material 18
Total 100
Ion exchange capacity (meq/100g) 193
Table 3. Chemical composition of Petrota natural zeolite sample ZeotP.
Table 3. Chemical composition of Petrota natural zeolite sample ZeotP.
Sample SiO2 Al2O3 Fe2O3tot MnO MgO CaO SrO BaO Na2O K2O H2O Total
ZeotP 68.43 11.60 bdl* bdl 0.96 3.80 bdl bdl 0.03 2.12 13.03 99.97
* bdl: below detection limit.
Table 4. Mean (±SE) number of B. oleae oviposition holes in olives immersed to 5% ZeotP aqueous solution, with and without NU-FILM-P® adjuvant. ZeotP treated and non-treated (control) olives were maintained in cages with 5 B. oleae females and the number of oviposition holes was determined after 3 and 6 days at a temperature of 25°C and a 16L:8D photoperiod.
Table 4. Mean (±SE) number of B. oleae oviposition holes in olives immersed to 5% ZeotP aqueous solution, with and without NU-FILM-P® adjuvant. ZeotP treated and non-treated (control) olives were maintained in cages with 5 B. oleae females and the number of oviposition holes was determined after 3 and 6 days at a temperature of 25°C and a 16L:8D photoperiod.
Product Mean (±SE) number of eggs (oviposition holes) after
3 6
days
Control 107.0 ± 8.4a* 168.5 ± 12.2a
ZeotP without NU-FILM-P® 16.2 ± 6.4ab 59.0 ± 15.7ab
ZeotP with NU-FILM-P® 0.0 ± 0.0b 0.0 ± 0.0b
* Means in a column followed by the same letter are not significantly different (P > 0.05).
Table 5. Mean (±SE) number of B. oleae oviposition holes on olives immersed to 5%/ ZeotP aqueous solution, with NU-FILM-P® adjuvant, at four (17, 20, 25, 30°C) constant temperatures. ZeotP treated and non-treated (control) olives were maintained in cages with 5 B. oleae females and the number of oviposition holes was determined after 3 and 6 days at a 16L:8D photoperiod.
Table 5. Mean (±SE) number of B. oleae oviposition holes on olives immersed to 5%/ ZeotP aqueous solution, with NU-FILM-P® adjuvant, at four (17, 20, 25, 30°C) constant temperatures. ZeotP treated and non-treated (control) olives were maintained in cages with 5 B. oleae females and the number of oviposition holes was determined after 3 and 6 days at a 16L:8D photoperiod.
Temperature Mean (±SE) number of eggs (oviposition holes) after
3 6
days
Control ZeotP with NU-FILM-P® Control ZeotP with NU-FILM-P®

17°C
13.5 ± 7.7ab*
0.0 ± 0.0e
35.0 ± 17.0ac
0.2 ± 0.2e
20°C 9.7 ± 2.9b 0.0 ± 0.0e 37.5 ± 10.5ac 0.0 ± 0.0e
25°C 81.5 ± 30.8d 0.2 ± 0.2e 129.0 ± 28.7b 0.7 ± 0.7e
30°C 49.2 ± 14.9cd 0.2 ± 0.2e 100.7 ± 22.2bd 1.2 ± 1.2ef
* Means in two consecutive columns followed by the same letter are not significantly different (P > 0.05).
Table 6. Mean (±SE) number of B. oleae oviposition holes in olives immersed to 5% ZeotP aqueous solution, with NU-FILM-P® adjuvant, at five different RH levels (23, 33, 55, 75 and 94%). ZeotP treated and non-treated (control) olives were maintained in cages with 5 B. oleae females and the number of oviposition holes was determined after 3 and 6 days, at a temperature of 25°C and a 16L:8D photoperiod.
Table 6. Mean (±SE) number of B. oleae oviposition holes in olives immersed to 5% ZeotP aqueous solution, with NU-FILM-P® adjuvant, at five different RH levels (23, 33, 55, 75 and 94%). ZeotP treated and non-treated (control) olives were maintained in cages with 5 B. oleae females and the number of oviposition holes was determined after 3 and 6 days, at a temperature of 25°C and a 16L:8D photoperiod.
RH % Mean (±SE number of eggs (oviposition holes) after
3 6
days
Control ZeotP with NU-FILM-P® Control ZeotP with NU-FILM-P®
23 33.2 ± 17.0ac* 4.7 ± 2.7d 47.5 ± 21.4ab 9.5 ± 0.6d
33 34.5 ± 10.4ab 9.2 ± 1.1df 69.5 ± 8.3bc 10.0 ± 1.1de
55 73.2 ± 10.2c 4.2 ± 1.9d 84.2 ± 5.6bc 7.2 ± 3.1d
75 60.7 ± 8.4ce 3.0 ± 2.3d 90.5 ± 2.7c 6.5 ± 3.1d
94 27.0 ± 3.3bd 7.0 ± 1.1df 43.0 ± 3.4a 11.0 ± 1.1de
*Means in two consecutive columns followed by the same letter are not significantly different (P > 0.05).
Table 7. Residual oviposition deterrent effect of ZeotP after spraying with water the treated olive fruit. Olives were immersed to 5% ZeotP with or without NU-FILM-P® and subsequently sprayed with 5 and 15 ml of water. Then, the olives were maintained in cages (5 olives with 5 B. oleae females) and the number of oviposition holes was determined after 3, 6 and 9 days. Temperature 25°C, 16L:8D photoperiod.
Table 7. Residual oviposition deterrent effect of ZeotP after spraying with water the treated olive fruit. Olives were immersed to 5% ZeotP with or without NU-FILM-P® and subsequently sprayed with 5 and 15 ml of water. Then, the olives were maintained in cages (5 olives with 5 B. oleae females) and the number of oviposition holes was determined after 3, 6 and 9 days. Temperature 25°C, 16L:8D photoperiod.
Product Mean (±SE) number of eggs (oviposition holes) after
3 6 9
days
Water
L Volume
(ml)		 With
NU-FILM-P® 		Without
NU-FILM-P® 		With
NU-FILM-P® 		Without
NU-FILM-P® 		With
NU-FILM-P® 		Without
NU-FILM-P®
ZeotP 0 0.0 ± 0.0a* 7.5 ± 4.1ab 0.0 ± 0.0a 39.7 ± 12.8ac 0.0 ± 0.0a 56.5 ± 14.1a
5 1.0 ± 0.7a 10.5 ± 6.1b 1.7 ± 0.5a 34.2 ± 19.9a 3.0 ± 1.1ab 42.3 ± 14.6b
15 0.0 ± 0.0a 13.8 ± 5.5b 0.0 ± 0.0a 41.8 ± 8.0b 1.8 ± 1.4a 60.8 ± 20.5bc
Decis® 0 2.8 ± 0.9ab 4.5 ± 1.8ab 3.0 ± 1.1ab 16.5 ± 7.0ab 4.7 ± 2.1b 32.2 ± 18.0b
5 1.0 ± 1.0a 13.5 ± 6.2b 1.5 ± 1.2a 36.2 ± 11.6b 6.2 ± 0.9c 66.2 ± 18.7c
15 3.0 ± 1.3a 0.8 ± 0.8a 4.5 ± 0.5b 4.5 ± 4.5d 4.5 ± 2.4bc 22.5 ± 10.7a
* Means in a column followed by the same letter are not significantly different (P > 0.05).
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.
Copyright: This open access article is published under a Creative Commons CC BY 4.0 license, which permit the free download, distribution, and reuse, provided that the author and preprint are cited in any reuse.
Prerpints.org logo

Preprints.org is a free preprint server supported by MDPI in Basel, Switzerland.

facebook logotwitter logolinkedin logo
bsky
MDPI Initiatives
Important Links
Subscribe

Disclaimer

Terms of Use

Privacy Policy

Privacy Settings

© 2026 MDPI (Basel, Switzerland) unless otherwise stated