The Effect of Different Forms of Titanium Dioxide on the Yield, Chemical and Microbiological Parameters of Perennial Ryegrass (Lolium perenne L.) Herbage and Silage
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Physicochemical Characteristics of TiO2NPs
2.3. Experimental Design
2.4. Chemical Analysis
2.5. Quantitative PCR
2.6. Statistical Analysis
3. Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Properties | TiO2NPs1 | TiO2NPs2 | TiO2Com |
---|---|---|---|
Concentration of stock suspension [mg L−1] | 100 | 100 | 8.5 |
Conductivity [μS cm−1] | 5.4 | 8.6 | 8.6 |
pH | 5.5 | 5.9 | 5.8 |
Diameter [nm] 1 | 68 ± 7 | 207 ± 17 | ND 3 |
Hydrodynamic diameter [nm] 2 | 812 ± 30 | 441 ± 40 | ND |
Electrophoreticmobility [(μmcm) (Vs)−1 ] 2 | 2.13 ± 0.04 | −3.74 ± 0.03 | ND |
Zeta potential [mV] 2 | 31 ± 1 | −58 ± 1 | ND |
Target (Quantitative PCR) | Primer/Probe Sets | Reaction Mix | Reaction Conditions | Plasmid Preparation (Standard) 1 | References |
---|---|---|---|---|---|
Total bacterial load/Lactobacillus spp. | F_eubR_eubP_eub (probe) F_alllact_ISR_alllact_IS P_alllact_IS (probe) | Total volume of 20 µL contained 0.5 µM of each primer, 0.2 µM of probe, 2 µM of isolated DNA sample and 10 µL of Maxima Probe qPCR Master Mix 2× (Thermo Fisher Scientific, Waltham, MA, USA). | Initial denaturation step of 95 °C for 10 min, followed by 42 cycles at 95 °C for 15 s and 60 °C for 1 min. | Bacillus subtilis Lactobacillus plantarum | Haarman and Knol [30] |
Clostridium spp. | CI-F1 CI-R2 Probe-I (probe) | Initial denaturation at 95 °C for 10 min, 45 cycles at 95 °C for 20 s, at 63 °C for 30 s, and at 72 °C for 45 s. | Clostridium perfringens | Song et al. [31] | |
Yeast | YEASTF YEASTR | Total volume of 20 µL contained 0.4 µM of each primer, 2 µM of isolated DNA sample and 10 µL of Maxima SYBR Green qPCR Master Mix 2× (Thermo Fisher Scientific, Waltham, MA, USA). | Initial denaturation at 95 °C for 10 min followed by 40 cycles of 95 °C for 15 s, 60 °C for 1 min, and 72 °C for 30 s. | Saccharomyces spp. | Hierro et al. [32] |
Fungal load | NSI1 58A2R | Initial denaturation at 95 °C for 10 min, 42 cycles at 95 °C for 15 s, 52°C for 30 s, 72 °C for 30 s, and fluorescent data collection at 78 °C for 15 s. Melting curve analyses were performed as follows: denaturation step at 95 °C for 15 s, annealing at 60 °C for 1 min and melting in 0.3 °C steps up to 95 °C for 15 s. | Fusarium culmorum | Martin and Rygiewicz [33] Hemkemeyer et al. [34] | |
Bacillus spp. | 16SBACF 16SBACR | Total volume of 20 µL contained 0.5 µM of each primer, 2 µM of isolated DNA sample and 10 µL of Maxima SYBR Green qPCR Master Mix 2× (Thermo Fisher Scientific, Waltham, MA, USA). | Initial denaturation at 95 °C for 10 min, 45 cycles at 95 °C for 15 s, at 58 °C for 55 s, and at 72 °C for 30 s. Melting curve analyses were performed as follows: denaturation step at 95 °C for 15 s, annealing at 60 °C for 1 min and melting in 0.3 °C steps up to 95 °C for 15 s. | Bacillus subtilis | Mora et al. [35] |
Toxigenic Fusarium spp. | Tox5-1 Tox5-2 | Initial denaturation at 95 °C for 10 min, 45 cycles at 95 °C for 10 s, at 63 °C for 10 s, and at 72 °C for 30 s. Melting curve analyses were performed as follows: denaturation step at 95 °C for 15 s, annealing at 60 °C for 1 min and melting in 0.3 °C steps up to 95 °C for 15 s. | Fuariumculmorum | Schnerr et al. [36] |
Treatment | Unit | CONT | TiO2NPs1 | TiO2NPs2 | TiO2Com | St dev. 1 | St. err | p-Value |
---|---|---|---|---|---|---|---|---|
Yield | Mg ha−1 | 12.2 ab | 13.2 a | 12.5 ab | 11.5 b | 0.971 | 0.402 | 0.026 |
DM 2 | g kg−1 | 317 | 329 | 318 | 332 | 11.80 | 59.34 | 0.370 |
Ash | g kg−1 DM | 85.5 | 96.1 | 83.1 | 78.6 | 13.57 | 78.56 | 0.495 |
CP | 127 | 121 | 127 | 116 | 8.020 | 27.44 | 0.234 | |
NDF | 498 | 492 | 472 | 488 | 19.74 | 166.1 | 0.461 | |
ADF | 330 ab | 331 a | 306 b | 315 a | 17.14 | 125.3 | 0.025 | |
WSC | 155 | 163 | 161 | 164 | 10.64 | 48.31 | 0.803 | |
Total bacteria | Gene copies g−1 DM | 2.2 × 106 d | 2.1 × 107 c | 3.5 × 107 b | 5.7 × 107 a | 2.1 × 107 | 6.1 × 106 | <0.0001 |
Lactobacillus spp. | 3.1 × 103 b | 4.0 × 103 b | 6.5 × 104 a | 1.3 × 103 c | 2.8 × 104 | 8.1 × 103 | <0.0001 | |
Clostridium spp. | 7.7 × 103 a | 1.2 × 102 c | 1.0 × 102 d | 1.5 × 102 b | 3.0 × 102 | 8.5 × 101 | <0.0001 | |
Total fungi | 1.6 × 107 b | 1.6 × 107 b | 1.9 × 107 a | 1.6 × 107 b | 1.3 × 106 | 3.7 × 105 | 0.005 | |
Yeast | 6.9 × 106 c | 7.1 × 106 b | 7.4 × 106 a | 6.0 × 106 d | 5.3 × 105 | 1.5 × 105 | <0.0001 | |
Bacillus spp. | 4.4 × 108 b | 8.6 × 107 c | 5.8 × 107 c | 7.6 × 108 a | 3.1 × 108 | 8.8 × 107 | <0.0001 | |
Penicillium spp. | 7.7 × 104 b | 1.2 × 105 ab | 8.2 × 104 b | 1.4 × 105 ab | 3.0 × 104 | 8.8 × 103 | 0.020 | |
Fusarium spp. | 2.6 × 101 a | 0 b | 0 b | 0 b | 1.3 × 101 | 4 | 0.003 |
Parameter | Unit | CONT | TiO2NPs1 | TiO2NPs2 | TiO2Com | St dev. 2 | St. err | p-Value |
---|---|---|---|---|---|---|---|---|
Dry matter | g kg−1 | 316 | 333 | 321 | 332 | 13.56 | 78.44 | 0.385 |
Ash | g kg−1 DM | 122 | 111 | 101 | 101 | 19.93 | 169.4 | 0.469 |
Crude protein | 122 ab | 124 ab | 129 a | 117 b | 5.623 | 13.48 | 0.046 | |
NDF 1 | 499 | 487 | 468 | 487 | 17.83 | 135.6 | 0.189 | |
ADF | 334 | 324 | 313 | 321 | 15.25 | 99.20 | 0.431 | |
WSC | 17.4 c | 18.8 bc | 22.0 ab | 24.9 a | 3.238 | 4.470 | 0.000 | |
pH | 4.68 a | 4.64 a | 4.49 b | 4.63 ab | 0.089 | 0.003 | 0.017 | |
N-NH3 | g kg−1 DM | 105 | 111 | 103 | 109 | 9.899 | 41.78 | 0.796 |
Lactic acid | 83.2 b | 94.8 a | 86.3 ab | 77.8 b | 7.086 | 21.41 | 0.002 | |
Acetic acid | 3.30 | 3.75 | 4.21 | 3.40 | 0.847 | 0.306 | 0.618 | |
Propionic acid | 0.06 | 0.05 | 0.04 | 0.04 | 0.034 | 0.000 | 0.948 | |
Ethanol | 1.09 | 1.31 | 2.83 | 0.46 | 2.142 | 1.956 | 0.643 | |
Butyric acid | 1.33 | 0.48 | 0.42 | 0.34 | 0.735 | 0.230 | 0.347 | |
Valeric acid | 0.03 | 0.03 | 0.03 | 0.04 | 0.011 | 0.000 | 0.529 | |
Total bacteria | Gene copies g−1 DM | 1.4 × 1010 | 8.7 × 109 | 5.5 × 109 | 8.4 × 109 | 4.8 × 109 | 1.39 × 109 | 0.270 |
Lactobacillus spp. | 1.33 × 109 | 1.3 × 109 | 1.5 × 109 | 9.2 × 108 | 3.8 × 108 | 1.1 × 108 | 0.450 | |
Clostridium spp. | 4.0 × 108 a | 6.8 × 108 a | 8.0 × 105 b | 3.3 × 106 b | 4.5 × 108 | 1.3 × 108 | 0.018 | |
Total fungi | 4.4 × 1010 | 4.8 × 109 | 6.7 × 109 | 4.2 × 109 | 1.1 × 107 | 3.1 × 106 | 0.697 | |
Yeast | 1.2 × 107 ab | 1.2 × 107 ab | 1.3 × 107 b | 8.9 × 106 a | 1.6 × 1011 | 4.7 × 1010 | 0.050 | |
Bacillus spp. | 6.4 × 107 | 5.2 × 107 | 5.7 × 107 | 4.7 × 107 | 5.7 × 107 | 1.6 × 107 | 0.285 | |
Penicillium spp. | 2.8 × 107 a | 3.0 × 106 b | 5.4 × 106 b | 3.3 × 106 ab | 8.7 × 107 | 2.5 × 107 | 0.038 | |
Fusarium spp. | 0 * | 0 | 0 | 0 | 1.7 × 1010 | 4.8 × 109 | 1.000 |
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Przemieniecki, S.W.; Borsuk-Stanulewicz, M.; Purwin, C.; Kosewska, O.; Oćwieja, M. The Effect of Different Forms of Titanium Dioxide on the Yield, Chemical and Microbiological Parameters of Perennial Ryegrass (Lolium perenne L.) Herbage and Silage. Agriculture 2023, 13, 1588. https://doi.org/10.3390/agriculture13081588
Przemieniecki SW, Borsuk-Stanulewicz M, Purwin C, Kosewska O, Oćwieja M. The Effect of Different Forms of Titanium Dioxide on the Yield, Chemical and Microbiological Parameters of Perennial Ryegrass (Lolium perenne L.) Herbage and Silage. Agriculture. 2023; 13(8):1588. https://doi.org/10.3390/agriculture13081588
Chicago/Turabian StylePrzemieniecki, Sebastian Wojciech, Marta Borsuk-Stanulewicz, Cezary Purwin, Olga Kosewska, and Magdalena Oćwieja. 2023. "The Effect of Different Forms of Titanium Dioxide on the Yield, Chemical and Microbiological Parameters of Perennial Ryegrass (Lolium perenne L.) Herbage and Silage" Agriculture 13, no. 8: 1588. https://doi.org/10.3390/agriculture13081588