Influence of Amitraz and Oxalic Acid on the Cuticle Proteolytic System of Apis mellifera L. Workers
Abstract
:1. Introduction
2. Results and Discussion
Control | Amitraz | Oxalic acid | |||
---|---|---|---|---|---|
Substrate | proteases | pH | Mean ± se | Mean ± se | Mean ± se |
Albumin | Hydrophilic (rinsed in water) | 2.4 | 2.17 ± 0.77 | 4.74 ± 3.21 | 2.55 ± 1.79 |
7 | 13.88 ± 6.82 | 8.63 ± 3.74 | 7.74 ± 3.42 | ||
11.2 | 21.889a ± 5.06 | 20.15a ± 4.61 | 10.52b ± 1.55 | ||
Hydrophobic (rinsed in triton) | 2.4 | 12.66b ± 0.03 | 13.74b ± 1.93 | 19.57a ± 4.72 | |
7 | 7.867 ± 0.05 | 14.53 ± 1.85 | 20.13 ± 5.12 | ||
11.2 | 167.71a ± 0.22 | 13.91b ± 1.77 | 19.54c ± 4.70 | ||
hemoglobin | Hydrophilic (rinsed in water) | 2.4 | 1.87 ± 0.87 | 1.00 ± 0.50 | 1.00 ± 1.00 |
7 | 1.73 ± 0.16 | 1.08 ± 0.43 | 1.79 ± 1.54 | ||
11.2 | 2.56a ± 0.33 | 0.44b ± 0.16 | 1.55ab ± 0.83 | ||
Hydrophobic(rinsed in triton) | 2.4 | 15.58a ± 0.14 | 5.67b ± 0.99 | 2.87c ± 0.71 | |
7 | 20.64a ± 0.07 | 5.22b ± 1.15 | 4.38b ± 1.17 | ||
11.2 | 10.72a ± 0.07 | 2.96b ± 1.23 | 3.29b ± 0.72 | ||
cytochrome C | Hydrophilic (rinsed in water) | 2.4 | 4.65a ± 0.61 | 0.15b ± 0.07 | 1.53c ± 0.15 |
7 | 7.42a ± 0.82 | 0.93b ± 0.11 | 0.68b ± 0.25 | ||
11.2 | 9.54a ± 0.31 | 2.19b ± 0.23 | 1.05c ± 0.11 | ||
Hydrophobic (rinsed in triton) | 2.4 | 12.27a ± 0.10 | 8.92b ± 1.15 | 12.33a ± 1.77 | |
7 | 6.03b ± 0.19 | 6.57a ± 0.57 | 6.71a ± 0.21 | ||
11.2 | 22.13a ± 0.16 | 6.66b ± 0.44 | 3.39c ± 0.26 | ||
ovoalbumin | Hydrophilic (rinsed in water) | 2.4 | 1.85a ± 0.16 | 0.51b ± 0.09 | 0.65c ± 0.22 |
7 | 14.54a ± 0.08 | 1.86b ± 0.36 | 0.00c ± 0.00 | ||
11.2 | 4.18a ± 0.21 | 0.00b ± 0.00 | 0.69b ± 0.17 | ||
Hydrophobic (rinsed in triton) | 2.4 | 51.18a ± 0.30 | 4.17b ± 0.31 | 2.72c ± 0.43 | |
7 | 23.71a ± 0.09 | 3.00b ± 0.49 | 2.178c ± 0.36 | ||
11.2 | 48.32a ± 0.22 | 3.02b ± 0.56 | 3.35b ± 0.53 |
2.1. Cuticle Proteases
Control—no treatment | Amitraz treatment | Oxalic acid treatment | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
hydrophilic protein | hydrophobic protein | hydrophilic protein | hydrophobic protein | hydrophilic protein | hydrophobic protein | ||||||||
pH | Rf (mm) | Band number | OD | Band number | OD | Band number | OD | Band number | OD | Band number | OD | Band number | OD |
2.4 | 0–40 | 1 | 0.25 | 3 | 0.86 | 3 | 0.21 | 4 | 0.25 | 3 | 0.15 | 1 | 0.20 |
41–60 | 1 | 0.40 | 2 | 0.35 | 1 | 0.26 | 3 | 0.43 | 1 | 0.35 | 1 | 0.23 | |
61–100 | 4 | 0.28 | 0 | 0 | 3 | 0.36 | 4 | 0.46 | 1 | 0.49 | 3 | 0.53 | |
7.0 | 0–40 | 5 | 0.18 | 2 | 0.54 | 1 | 0.49 | 4 | 0.66 | 0 | 0 | 3 | 0.48 |
41–60 | 2 | 0.20 | 0 | 0 | 1 | 0.63 | 1 | 0.92 | 2 | 0.47 | 1 | 0.74 | |
61–100 | 4 | 0.25 | 3 | 0.36 | 2 | 0.61 | 2 | 0.68 | 3 | 0.52 | 2 | 0.74 | |
11.0 | 0–40 | 3 | 0.25 | 0 | 0 | 1 | 0.30 | 5 | 0.23 | 1 | 0.12 | 2 | 0.14 |
41–60 | 0 | 0 | 0 | 0 | 1 | 0.23 | 1 | 0.15 | 1 | 0.19 | 1 | 0.20 | |
61–100 | 1 | 0.32 | 0 | 0 | 4 | 0.25 | 1 | 0.18 | 2 | 0.18 | 3 | 0.38 |
2.2. Cuticle Protease Inhibitors
Control—no treatment | Amitraz treatment | Oxalic acid treatment | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
hydrophilic protein | hydrophobic protein | hydrophilic protein | hydrophobic protein | hydrophilic protein | hydrophobic protein | ||||||||
pH | Rf (mm) | Band number | OD | Band number | OD | Band number | OD | Band number | OD | Band number | OD | Band number | OD |
2.4 | 0–40 | 4 | 0.21 | 0 | 0 | 5 | 0.73 | 2 | 0.74 | 4 | 0.65 | 5 | 0.63 |
41–60 | 0 | 0 | 1 | 0.20 | 1 | 0.58 | 1 | 0.67 | 1 | 0.55 | 0 | 0 | |
61–100 | 3 | 0.21 | 4 | 0.14 | 5 | 0.53 | 4 | 0.56 | 2 | 0.58 | 3 | 0.61 | |
7.0 | 0–40 | 2 | 0.16 | 1 | 0.23 | 2 | 0.23 | 4 | 0.35 | 3 | 0.26 | 1 | 0.23 |
41–60 | 0 | 0 | 0 | 0 | 1 | 0.30 | 2 | 0.52 | 0 | 0 | 0 | 0 | |
61–100 | 0 | 0 | 1 | 0.21 | 1 | 0.27 | 4 | 0.34 | 2 | 0.26 | 5 | 0.39 | |
11.0 | 0–40 | 3 | 0.37 | 4 | 0.23 | 2 | 0.14 | 2 | 0.16 | 2 | 0.08 | 0 | 0 |
41–60 | 0 | 0 | 0 | 0 | 2 | 0.10 | 0 | 0 | 1 | 0.11 | 0 | 0 | |
61–100 | 3 | 0.36 | 0 | 0 | 0 | 0 | 3 | 0.28 | 1 | 0.04 | 3 | 0.23 |
2.3. Apian Cuticle Proteolytic System — General Comments
2.4. Activity towards Model Microorganisms
Group | Antifungal activity | Antibacterial activity | ||||
---|---|---|---|---|---|---|
Candida albicans | Aspergillus niger | Bacillus subtilis | Staphylococcus aureus | Salmonella typhimurium | Pseudomonas aeruginosa | |
Control | 978.57 | 23.36 | 78.36 | 308.61 | 126.49 | 101.33 |
Amitraz | 212.50 | 0 | 756.51 | 0 | 259.62 | 207.24 |
Oxalic acid | 340.69 | 0 | 154.18 | 0 | 0 | 0 |
3. Experimental Section
3.1. The Experimental Design
3.2. Sampling and the Data Base
- 2 mL—for determining protease and protease inhibitor activities (portion a),
- 2 mL—for electrophoretic assays (portion b),
- 2 mL—for determining antifungal and antibacterial activities in vivo (portion c),
- 2 mL—reserve (portion d).
3.3. The Analytical Procedures
- the general protein content by the Lowry method, as modified by Schacterle and Pollack [31];
- the proteolytic activity in relation to the diagnostic inhibitors of proteolytic enzymes (pepstatin A, PMSF, iodoacetamide, o-phenanthroline), by the Lee and Lin method [33];
- the activities of acidic, neutral, and alkaline proteases according to the Anson method [32];
- the levels of natural inhibitors of acidic, neutral, and alkaline proteases, based on the Lee and Lin method [33].
3.4. In Vivo Laboratory Tests
- SABG [36]—to determine activity in relation to Aspergillus niger,
- YPD [37]—to determine activity in relation to Candida albicans,
- LB [38]—to determine activity in relation to Staphylococcus ureus (ATCC 25923), Bacillus subtilis (ATCC 6633), Micrococcus luteus (ATCC 7468), Salmonella typhimurium (ATCC 13311), Pseudomonas aeruginosa (ATCC 17853), and Escherichia coli (ATCC 10536).
3.5. Statistics
4. Conclusion
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Strachecka, A.; Paleolog, J.; Olszewski, K.; Borsuk, G. Influence of Amitraz and Oxalic Acid on the Cuticle Proteolytic System of Apis mellifera L. Workers. Insects 2012, 3, 821-832. https://doi.org/10.3390/insects3030821
Strachecka A, Paleolog J, Olszewski K, Borsuk G. Influence of Amitraz and Oxalic Acid on the Cuticle Proteolytic System of Apis mellifera L. Workers. Insects. 2012; 3(3):821-832. https://doi.org/10.3390/insects3030821
Chicago/Turabian StyleStrachecka, Aneta, Jerzy Paleolog, Krzysztof Olszewski, and Grzegorz Borsuk. 2012. "Influence of Amitraz and Oxalic Acid on the Cuticle Proteolytic System of Apis mellifera L. Workers" Insects 3, no. 3: 821-832. https://doi.org/10.3390/insects3030821