Growing Lettuce (Lactuca sativa L.) in Floating Disk Systems Under Variable and High Salinity Ranges in Water Enriched with Nanobubbles †
by
,
Konstantinos Zoukidis
1,2, 
Anastasia Giannakoula
2,
Ramonna Kosheleva
3,
Athanasios Gertsis
1,*, 
Antonios Apostolidis
1,
Georgios Strouthopoulos
1 and
Athanasios Varoutoglou
3 1
Department of Sustainable Agriculture and Management, American Farm School, Perrotis College, 54 Marinou Antypa Street, 57001 Thessaloniki, Greece
2
Department of Agriculture, International Hellenic University, 57400 Thessaloniki, Greece
3
Hephaestus Lab, Department of Chemistry, Democritus University of Thrace, 65404 Kavala, Greece
*
Author to whom correspondence should be addressed.
†
Presented at the 11th International Conference on Information and Communication Technologies in Agriculture, Food & Environment, Samos, Greece, 17–20 October 2024.
Proceedings 2025, 117(1), 33; https://doi.org/10.3390/proceedings2025117033
Published: 16 June 2025
(This article belongs to the Proceedings of 11th International Conference on Information and Communication Technologies in Agriculture, Food and Environment (HAICTA 2024), Volume II)
Abstract
:Hydroponic systems, which use commercial hydroponics technologies, are cheaper and easier to maintain than traditional farming methods in soil. The objective of this study was to evaluate various salinity ranges (E.C.i from 1 dS/m to 14 dS/m) in water enriched with nanobubbles (NBs) for the growth and productivity of lettuce plants in a floating disk hydroponic system. This research study investigated how using floating disks in a greenhouse with a nanobubble (NB) generator may affect lettuce’s (Lactuca sativa L.) morphological and physiological responses to salt stress. The goal of this experiment was to examine the results of the influence of NB and non-NB treatments on agronomic traits and yield. The results indicated that the NB device is an innovative and very effective technology for sustainable lettuce production under a high-salinity nutrient solution. This device presents a valuable solution to the global issue of the increased salinity of irrigation water.
1. Introduction
Lettuce (Lactuca sativa L.) is one of the world’s most commonly consumed vegetable horticultural crops [1]. The many natural health-promoting phytochemicals and vitamins found in lettuce include glycosylated flavonoids, hydroxycinnamic acids, sesquiterpene lactones, carotenoids, B family vitamins, ascorbic acid, and tocopherols. The secondary metabolites in lettuce have been linked to various health benefits, such as protection against cancer, diabetes, inflammation, and heart disease [2].
The salt concentration in full-strength seawater causes hazardous effects on most plants. It destroys soil structure, particularly without an adequate leaching mechanism, making large-scale sustainable agriculture dependent on seawater impossible [3]. Soil salinity significantly impacts lettuce’s fresh production and plant water consumption up to 2.17 dS/m, after which its impact levels off. The optimal salinity for iceberg lettuce plants is 1.84 dS/m, with relative yield reductions of 8.26 and 22.7% per unit salinity increase above and below the optimum level [4]. While lettuce plants cannot absorb as much water in salty environments, their morphology and histology change, leading to effects like thicker leaves, which alter the plant’s overall texture [5]. Osmotic stress, resulting from salt accumulation in the root zone, reduces water intake. Reduced water absorption slows transpiration, cell division and enlargement, metabolic activity, and development [6].
The lifetime of nanobubbles is very extended, lasting weeks or even months. Nanobubbles (NB) are gas cavities with a diameter of 1 μ or less that exist on the nanoparticle scale [7]. The incredible lifespan and the unique physicochemical properties of NB structures have piqued the scientific community’s interest [8]. The mechanisms and processes underlying this trait have been the subject of much research. Many industries, including agriculture, have taken notice of nanotechnology and nanobubbles because of their expanding importance and unique qualities [9].
Based on the principles of commercial hydroponics, SSC systems cost less money and effort to maintain than traditional farming methods [10]. There are a variety of SSC setups available, and they vary most noticeably in their choice of building materials, plant growth substrate, and nutrient solution control [11].
The objective of this study was to evaluate various water salinity ranges in irrigation water (E.C.i from 1 dS/m to 14 dS/m) enriched with nanobubbles (NB) for the growth and productivity of lettuce plants in a floating disk hydroponic system.
2. Materials and Methods
This study took place in a greenhouse at Perrotis College/American Farm School, Thessaloniki, Greece. This study was conducted from 20 November 2023 to 22 February 2024. More growing cycles with other varieties of lettuce (COS, red leaf, and butterhead) are in progress.
In this experimental design, a floating disk hydroponic cultivation system was chosen, and the Batavia Epsilone lettuce variety was used. The lettuce was in plastic containers (6 plants per plastic container) of 30 L volume (Figure 1) filled with irrigation water with nutrient solution (50% of Hoagland solution). We performed different treatments, which were as follows: The first part of experiment was the control (not enriched with NBs) group, for which we filled 18 plastic containers with regular tap water/nutrient solution with a range of salinity concentrations (E.C.i: ~1, 2, 4, 6, 8, and 10 dS/m). In the second part of the experiment, the lettuce was grown in 24 plastic containers with a range of salinity concentrations in water (E.C.i: ~1, 2, 4, 6, 8, 10, 12, and 14 dS/m) enriched with NBs every 3 days [12]. In the third part of the experiment, the lettuce was grown in 4 plastic containers that were enriched with NBs (E.C.i: ~1 dS/m) every 7 days, every 3 days, every day, and just once.
Various vegetable agronomic parameters (total fresh green weight and root weight and length), NDVI (Normalized Difference Vegetation Index) (measured by the Trimble GreenSeeker Handheld Crop Sensor, Trimble Inc., Westminster, CO, USA), and Relative Leaf Chrolophyll Levels (measured by SPAD 502 Minolta, KONICA MINOLTA, Inc. Tokyo, Japan) were measured at the end of the experiment, and water parameters (Dissolved Oxygen, pH, EC, temperature, and the size and concentration of NBs) were also periodically recorded. Measurements of the environmental conditions inside in the greenhouse such as air temperature (°C) and air humidity (%), were continuously recorded from a portable meteorological station.
The measured data were statistically analyzed for mean differences between the treatments using Student’s t test, since the data were normally distributed, and any outlier values were excluded. The statistical software JMP v 18 was used (www.jmp.com accessed on 2 April 2024).
3. Results and Discussion
Our study of Batavia lettuce showed very good results. Most of the treatments of Batavia lettuce (E.C.i: 1, 2, 4, 6, 8, 10, and 12 dS/m) enriched with NBs showed the maximum fresh green biomass weight (Table 1) and demonstrated a statistically significant difference from the regular tap water treatment (E.C.i: ~1, 2, 4, 6, 8, and 10 dS/m). In Table 1, it can be seen that the treatments (E.C.i: 2 and 4 dS/m) produced the maximum fresh green root weight. The results for root length reflected the same trend shown for fresh green root weight (Table 1). Relative Leaf Chlorophyll Level (SPAD units, Table 2) and chlorophyll fluorescence emission (PSII) (Table 2) confirmed the trend shown for the fresh weight of lettuce.
The generator of AgroNBs enriched with NBs every 3 days had the same results as the NB enrichment treatment performed every day, and the two treatments did not demonstrate a statistically significant difference in terms of all the agronomic traits measured [fresh green biomass and root weight and fresh green biomass and root length (Table 3)]. Furthermore, some other agronomic properties measured, such as NDVI (Normalized Difference Vegetation Index) (Table 4), Relative Leaf Chlorophyll Level (SPAD units) (Table 4), and chlorophyll fluorescence emission (PSII) (Table 4), confirmed the trend shown for the previous agronomic traits. The results of this study are confirmed by the results of other researchers [13].
The generator of AgroNBs produced the best/optimum size (Table 5) of NBs (200 nm) in plastic containers filled with regular tap water/nutrient solution E.C.i (2 dS/m) enriched with NBs (290.5 nm), and the maximum concentration (Table 5) of NB (106) particles, measured in nanobubbles/mL, was produced in plastic containers filled with regular tap water/nutrient solution with high E.C.i (10 dS/m) and E.C.i (12 dS/m) enriched with NBs.
4. Conclusions and Recommendations
The results show that NB-enriched treatments increased the growth and other productivity characteristics of lettuce almost directly proportionally with ECi. Tap water results showed the opposite effect with increasing ECi level. The NB device is highly recommended for floating disk production systems if the water used has a prohibitive value of EC and the plant species grown are sensitive to elevated EC values. Complimentary studies are in progress to test the device under soil-grown vegetables in pots and in the field.
Author Contributions
Conceptualization, K.Z. and A.G. (Athanasios Gertsis); methodology, K.Z., A.G. (Athanasios Gertsis) and A.G. (Anastasia Giannakoula); software, A.G. (Athanasios Gertsis) and K.Z.; validation, K.Z., A.G. (Athanasios Gertsis) and A.G. (Anastasia Giannakoula); formal analysis, K.Z. and A.G. (Athanasios Gertsis); investigation, K.Z., A.G. (Athanasios Gertsis), A.G. (Anastasia Giannakoula), R.K. and A.V.; resources, K.Z., A.G. (Athanasios Gertsis), R.K. and A.V.; data curation, K.Z., A.G. (Athanasios Gertsis), R.K., A.A. and G.S.; writing—original draft preparation, K.Z. and A.G. (Athanasios Gertsis); writing—review and editing, K.Z. and A.G. (Athanasios Gertsis); visualization, K.Z., A.G. (Athanasios Gertsis) and A.G. (Anastasia Giannakoula); supervision, K.Z., A.G. (Athanasios Gertsis) and A.G. (Anastasia Giannakoula); project administration, K.Z., A.G. (Athanasios Gertsis) and A.A.; funding acquisition, K.Z., A.G. (Athanasios Gertsis) and A.A. All authors have read and agreed to the published version of the manuscript.
Funding
This research received no external funding.
Institutional Review Board Statement
Not applicable.
Informed Consent Statement
Not applicable.
Data Availability Statement
The data presented in this study are available on request from the corresponding author due to ongoing analyses and planned future publications.
Acknowledgments
The authors express their sincere gratitude to the two companies that provided the generator of Agro-nanobubbles (NBs) (Hephaestus Lab, Department of Chemistry, Democritus University of Thrace) and the lettuce plants from nurseries of Sporofyta Vasilikon in Greece.
Conflicts of Interest
The authors declare no conflicts of interest.
Abbreviations
The following abbreviations are used in this manuscript:
| NBs | Nanobubbles |
| NDVI | Normalized Difference Vegetation Index |
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Figure 1.
Experimental design: layout of the treatments in plastic containers with variable- and high-salinity water enriched with NBs.
Figure 1.
Experimental design: layout of the treatments in plastic containers with variable- and high-salinity water enriched with NBs.
Figure 2.
Experimental design: layout of the treatments in plastic containers with variable- and high-salinity water (control; no NBs).
Figure 2.
Experimental design: layout of the treatments in plastic containers with variable- and high-salinity water (control; no NBs).
Figure 3.
Experimental design: layout of the treatments in plastic containers with different days of NB application.
Figure 3.
Experimental design: layout of the treatments in plastic containers with different days of NB application.
Figure 4.
A comprehensive picture of all treatments with the actual pants shown: plot plan.
Table 1.
Various vegetable agronomic parameters of Batavia lettuce (g/plant) by water treatment.
| Fresh Green Biomass (g/plant) | Fresh Root Weight (g/plant) | Fresh Root Length (cm/plant) | |
|---|---|---|---|
| Average and ± Standard Deviation Weight * | |||
| Main Treatment | Batavia Lettuce | ||
| Control, tap water (E.C. < 1 dS/m) | 12.8 Ɨ ± 3.4 ŧ ab * | 3.57 ± 1.6 abc | 11.59 ± 1.8 a |
| Control, tap water (E.C. 2 dS/m) | 4.29 ± 0.8 cd | 4.09 ± 2.3 a | 9.81 ± 1.3 ab |
| Control, tap water (E.C. 4 dS/m) | 4.37 ± 1.3 cd | 3.68 ± 0.9 ab | 7.60 ± 1.3 b |
| Control, tap water (E.C. 6 dS/m) | 4.4 ± 1.4 cd | 2.44 ± 2.0 ef | 3.14 ± 1.2 c |
| Control, tap water (E.C. 8 dS/m) | 3.50 ± 1.6 cd | 1.68 ± 1.0 f | 2.93 ± 1.1 c |
| Control, tap water (E.C. 10 dS/m) | 0.0 ± 0.0 d | 0.00 ± 0.0 g | 0.00 ± 0.0 c |
| Tap water (E.C. < 1 dS/m) + NBs | 14.24 ± 1.1 a | 3.36 ± 0.5 abcd | 11.58 ± 1.8 a |
| Tap water (E.C. 2 dS/m) + NBs | 15.72 ± 2.9 a | 3.04 ± 1.2 bcde | 11.89 ± 1.3 a |
| Tap water (E.C. 4 dS/m) + NBs | 15.93 ± 2.2 a | 2.95 ± 1.1 bcde | 11.75 ± 1.4 a |
| Tap water (E.C. 6 dS/m) + NBs | 16.17 ± 2.2 a | 2.78 ± 1.1 cde | 10.86 ± 1.3 ab |
| Tap water (E.C. 8 dS/m) + NBs | 16.23 ± 2.8 a | 2.27 ± 0.9 ef | 11.05 ± 1.2 ab |
| Tap water (E.C. 10 dS/m) + NBs | 16.84 ± 1.4 a | 2.39 ± 0.8 ef | 10.65 ± 1.7 ab |
| Tap water (E.C. 12 dS/m) + NBs | 15.5 ± 5.4 a | 2.38 ± 0.7 ef | 10.30 ± 1.0 ab |
| Tap water (E.C. 14 dS/m) + NBs | 7.55 ± 3.1 bc | 2.41 ± 1.2 def | 7.29 ± 1.7 b |
* The values are the means of 18 plants/main treatment; different letters in the same column indicate statistically significant differences among all main treatments, according Oneway Analysis-Student’s t test independent samples t-test and one-way ANOVA test based on gender, at a level of significance of 5% (p < 0.05) in the overall comparison. Ɨ: Average. ŧ: STDEV.
Table 2.
Relative Leaf Chlorophyll Level (SPAD units) and chlorophyll fluorescence emission (PSII) of Batavia lettuce by water treatment.
Table 2.
Relative Leaf Chlorophyll Level (SPAD units) and chlorophyll fluorescence emission (PSII) of Batavia lettuce by water treatment.
| Chlorophyll Fluorescence Emission (PSII) | Chlorophyll (SPAD Units) | |
|---|---|---|
| Average and ± Standard Deviation * | ||
| Main Treatment | Batavia Lettuce | |
| Control, tap water (E.C. < 1 dS/m) | 0.80 ± 1.4 a * | 22.65 ± 0.0 a |
| Control, tap water (E.C. 2 dS/m) | 0.79 ± 1.9 a | 23.3 ± 0.1 a |
| Control, tap water (E.C. 4 dS/m) | 0.77 ± 1.3 a | 19.45 ± 0.2 abc |
| Control, tap water (E.C. 6 dS/m) | 0.73 ± 1.1 b | 17.89 ± 0.1 bc |
| Control, tap water (E.C. 8 dS/m) | 0.69 ± 1.2 b | 14.96 ± 0.1 c |
| Control, tap water (E.C. 10 dS/m) | 0.00 ± 0.0 c | 0.00 ± 0.0 d |
| Tap water (E.C. < 1 dS/m) + NBs | 0.81 ± 1.1 a | 22.60 ± 0.1 ab |
| Tap water (E.C. 2 dS/m) + NBs | 0.80 ± 1.5 a | 21.30 ± 0.1 ab |
| Tap water (E.C. 4 dS/m) + NBs | 0.78 ± 1.7 a | 21.05 ± 0.0 ab |
| Tap water (E.C. 6 dS/m) + NBs | 0.79 ± 0.9 a | 21.38 ± 0.0 ab |
| Tap water (E.C. 8 dS/m) + NBs | 0.80 ± 1.2 a | 20.09 ± 0.0 ab |
| Tap water (E.C. 10 dS/m) + NBs | 0.78 ± 1.2 a | 19.13 ± 0.0 abc |
| Tap water (E.C. 12 dS/m) + NBs | 0.80 ± 1.1 a | 19.43 ± 0.0 abc |
| Tap water (E.C. 14 dS/m) + NBs | 0.77 ± 1.7 a | 19.05 ± 0.0 abc |
* The different letters indicate a significant (p < 0.05) difference. Data are presented as the mean ± STDEV of 18 replicates (plants/main treatment).
Table 3.
Various vegetable agronomic parameters of Batavia lettuce (g/plant) by main treatment.
| Fresh Green Biomass (g/plant) | Fresh Green Biomass Length (cm/plant) | Fresh Root Weight (g/plant) | Fresh Root Length (cm/plant) | |
|---|---|---|---|---|
| Main Treatment | Batavia Lettuce | |||
| NB enrichment once per day (E.C. < 1 dS/m) | 16.5 ± 2.2 a * | 8.83 ± 1.0 a | 3.23 ± 0.8 a | 25.33 ± 1.6 a |
| NB enrichment once per 3 days (E.C. < 1 dS/m) | 16.45 ± 0.9 a | 8.00 ± 1.5 a | 2.95 ± 0.2 a | 30.00 ± 1.7 ab |
| NB enrichment once per 7 days (E.C. < 1 dS/m) | 12.93 ± 2.9 b | 7.50 ± 1.9 a | 1.43 ± 1.9 b | 18.67 ± 1.7 b |
| NB enrichment only once (E.C. < 1 dS/m) | 11.73 ± 1.6 b | 5.00 ± 1.1 b | 0.97 ± 1.1 b | 10.50 ± 1.5 c |
* Means with the same letter are not significantly different at a level of significance of 5% (p < 0.05) in the overall comparison.
Table 4.
Relative Leaf Chlorophyll Level (SPAD units), chlorophyll fluorescence emission (PSII), and NDVI of Batavia lettuce by main treatment.
Table 4.
Relative Leaf Chlorophyll Level (SPAD units), chlorophyll fluorescence emission (PSII), and NDVI of Batavia lettuce by main treatment.
| Chlorophyll (SPAD Units) | Chlorophyll Fluorescence Emission (PSII) | NDVI | |
|---|---|---|---|
| Main Treatment | Batavia Lettuce | ||
| NB enrichment once per day (E.C. < 1 dS/m) | 19.2 ± 1.4 a * | 0.84 ± 0.0 a | 0.41 ± 0.0 a |
| NB enrichment once per 3 days (E.C. < 1 dS/m) | 18.9 ± 1.3 a | 0.84 ± 0.0 a | 0.39 ± 0.0 a |
| NB enrichment once per 7 days (E.C. < 1 dS/m) | 12.5 ± 1.8 b | 0.81 ± 0.0 ab | 0.36 ± 0.0 a |
| NB enrichment only once (E.C. < 1 dS/m) | 7.2 ± 1.5 c | 0.80 ± 0.0 b | 0.29 ± 0.1 b |
* Means with the same letter are not significantly different at a level of significance of 5% (p < 0.05) in the overall comparison.
Table 5.
Size (nm) of NBs and concentration of NB particles (10^6)/ml by salinity treatment.
| NB Size (nm) | Concentration of NB Particles (106)/mL | |
|---|---|---|
| Main Treatment | Batavia Lettuce | |
| Tap water (E.C. < 1 dS/m) + NBs | 444.2 ± 6.2 a * | 5.84 ± 1.8 b |
| Tap water (E.C. 2 dS/m) + NBs | 290.5 ± 1.0 d | 7.5 ± 0.8 b |
| Tap water (E.C. 4 dS/m) + NBs | 338.3 ± 1.4 cd | 8.79 ± 1.5 b |
| Tap water (E.C. 6 dS/m) + NBs | 339.8 ± 5.3 cd | 10.24 ± 0.7 b |
| Tap water (E.C. 8 dS/m) + NBs | 351.6 ± 4.7 bcd | 12.72 ± 6.8 ab |
| Tap water (E.C. 10 dS/m) + NBs | 417.6 ± 4.6 ab | 17.94 ± 4.6 a |
| Tap water (E.C. 12 dS/m) + NBs | 406.9 ± 2.2 abc | 18.59 ± 4.7 a |
* Means with the same letter are not significantly different at a level of significance of 5% (p < 0.05) in the overall comparison.
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MDPI and ACS Style
Zoukidis, K.; Giannakoula, A.; Kosheleva, R.; Gertsis, A.; Apostolidis, A.; Strouthopoulos, G.; Varoutoglou, A. Growing Lettuce (Lactuca sativa L.) in Floating Disk Systems Under Variable and High Salinity Ranges in Water Enriched with Nanobubbles. Proceedings 2025, 117, 33. https://doi.org/10.3390/proceedings2025117033
AMA Style
Zoukidis K, Giannakoula A, Kosheleva R, Gertsis A, Apostolidis A, Strouthopoulos G, Varoutoglou A. Growing Lettuce (Lactuca sativa L.) in Floating Disk Systems Under Variable and High Salinity Ranges in Water Enriched with Nanobubbles. Proceedings. 2025; 117(1):33. https://doi.org/10.3390/proceedings2025117033
Chicago/Turabian StyleZoukidis, Konstantinos, Anastasia Giannakoula, Ramonna Kosheleva, Athanasios Gertsis, Antonios Apostolidis, Georgios Strouthopoulos, and Athanasios Varoutoglou. 2025. "Growing Lettuce (Lactuca sativa L.) in Floating Disk Systems Under Variable and High Salinity Ranges in Water Enriched with Nanobubbles" Proceedings 117, no. 1: 33. https://doi.org/10.3390/proceedings2025117033
APA StyleZoukidis, K., Giannakoula, A., Kosheleva, R., Gertsis, A., Apostolidis, A., Strouthopoulos, G., & Varoutoglou, A. (2025). Growing Lettuce (Lactuca sativa L.) in Floating Disk Systems Under Variable and High Salinity Ranges in Water Enriched with Nanobubbles. Proceedings, 117(1), 33. https://doi.org/10.3390/proceedings2025117033
