Comparative Study on the Resistance of Beta-Cypermethrin Nanoemulsion and Conventional Emulsion in Blattella germanica
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagent
2.2. Breeding of Sensitive and Resistant Lines
2.3. Determination of LD50 and Calculation of Its Resistance Factor
2.4. Determination of Enzyme Activity
2.4.1. Preparation of Enzyme Solution
- (1)
- Preparation of AchE enzyme solution: Blattella germanica specimens, having fasted for 24 h post-application, were washed with flowing distilled water for 2 min, and then gently dried on filter paper. The head was isolated, immersed in phosphate buffer (pH = 8.0, 1/15 mol/L) containing 0.5% TritonX-100, homogenized in an ice bath, and centrifuged at 4000× g r/min for 15 min. The resulting supernatant was collected as the enzyme solution.
- (2)
- Preparation of GST enzyme solution: Following a 24-h fasting period post-application, Blattella germanica specimens were washed with flowing distilled water for 2 min and gently dried on filter paper. At 4 °C, the specimens were dissected, the food within the digestive tract was removed, and an enzyme solution was prepared from the midgut. This solution was created by homogenizing in phosphate buffer (pH 6.5, 0.1 mol/L) in an ice bath and subsequent centrifugation at 10,000× g r/min for 15 min. The supernatant was then collected as the enzyme solution.
- (3)
- Preparation of P450-O demethylase solution: Similar to the previous steps, Blattella germanica specimens that fasted for 24 h post-application were washed with flowing distilled water for 2 min and gently dried on filter paper. They were dissected at 4 °C, with food removed from the digestive tract. The enzyme solution was prepared from the midgut by adding phosphate-buffered solution (pH 7.8, 0.1 mol/L, containing 1 mmol/L EDTA, 1 mmol/L DTT, and 1 mmol/L PMSF) in an ice bath, followed by centrifugation at 10,000× g r/min for 15 min. The supernatant was collected as the enzyme solution.
2.4.2. Determination of Enzyme Activity
- (1)
- AchE activity was determined according to Groun’s modified Ellman method [13]. The final reaction system volume was 0.2 mL. During this process, 100 μL of 1/15 mol/L phosphate buffer (pH 8.0), 50 μL of 0.75 mmol/L substrate (thioacetylcholine iodide), and 50 μL of enzyme solution (adjusted to a protein content of 40~80 μg/mL) were mixed and allowed to react at 30 °C for 15 min. Subsequently, 1.8 mL of DTNB reagent was added, and colorimetric determination was carried out at a 412 nm wavelength.
- (2)
- GST activity was determined as previously described [14]. This involved adding 100 μL of enzyme solution to the reaction system, composed of 2.5 mL of phosphate buffer (pH 6.5, 0.1 mol/L), 0.1 mL of reduced GSH (100 mmol/L), and 20 μL of 2,4-dinitrobenzene (CDNB) acetone solution (50 mmol/L). The mixture was thoroughly combined and left at 25 °C for 20 min. After adding 0.5 mL of SDS (2.5%) and thorough mixing, the absorbance value at 340 nm was recorded using an ultraviolet spectrophotometer every 1 min for a total of 3 min. The enzyme activity [m OD/(mg·min)] was expressed based on the reaction rate, calculated from the absorbance change within 3 min.
- (3)
- Determination of P450-O demethylase activity [15]: For P450-O demethylase activity determination [15], 100 μL of 2.0 mmol/L p-nitroanisole (P-NA), 10 μL of 9.6 mmol/L reduced coenzyme II (NADPH), and 90 μL of enzyme solution were added to each well of a 96-well microtiter plate. The optical density value at 412 nm wavelength was recorded every 25 s for a total of 10 min using a microplate reader. The enzymatic reaction stage was maintained at 30 °C. The reaction rate was calculated based on the optical density change within the range of 0~0.2, and the enzyme activity [nOD/(mg·min)] was expressed as the reaction rate.
2.4.3. Determination of Protein Concentration
2.5. Data Processing
3. Results and Analysis
3.1. Comparison of Resistance Factor of Different Resistant Strains of Two Insecticides
3.2. Comparison of AchE Activity in Blattella germanica Strains with Varied Resistance to Two Insecticides
3.3. Comparison of the Effect of GST Enzyme of Blattella germanica Strains with Different Resistance to Two Insecticides
3.4. Comparison of the Effects of P450-O Demethylase in Blattella germanica Strains with Varied Resistance to Both Insecticides
3.5. Reference Value of Enzyme Activity in Blattella germanica Resistant to Emulsion by Biochemical Method
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Jiang, Z.K.; Zhao, X.Z.; Yin, X.Y. Efficacy of beta-cypermethrin against health pests. Med. Anim. Control 1992, 8, 76–78. (In Chinese) [Google Scholar]
- Wang, Y. WHO recommended formulation of sanitary pesticides for controlling important vectors and its application. Pestic. Sci. Manag. 2021, 27, 35–43. (In Chinese) [Google Scholar]
- Zeng, X.; Yu, C.; Gao, X. Biochemical characteristics of cytochrome P450 in resistant and susceptible strains of blattella germanica. China J. Vector Biol. Control 2004, 15, 175–179. (In Chinese) [Google Scholar]
- Agosin, M. Role of microsomal oxidations in insecticide degradation. In Comprehensive Insect Physiology, Biochemistry and Pharmacology; Kerkut, G.A., Gilbert, L.I., Eds.; Insect Control: Oxford, UK, 1985; Volume 12, pp. 647–712. [Google Scholar]
- Bureau of Disease Control and Prevention, Ministry of Health. Manual of Malaria Control; People’s Medical Publishing House: Beijing, China, 2007; pp. 83–85. (In Chinese)
- Zeng, L.; Sun, D.; Zhao, W. Resistance of Culex pipiens quinquefasciatus and Anopheles dirus to pyrethroid insecticides. China J. Vector Biol. 2020, 19, 505–506. (In Chinese) [Google Scholar]
- Wu, Y.; Shen, J.; You, Z. Breeding of resistant and susceptible strains of Helicoverpa armigera to fenvalerate. Acta Entomol. Sin. 1994, 37, 129–135. (In Chinese) [Google Scholar]
- Ma, L.; Dou, F.; Cao, R. Investigation on resistance of Musca domestica and its control strategy in Chengdu City. Med. Anim. Control 2021, 4, 171–172. (In Chinese) [Google Scholar]
- Xu, Z.; Cao, L. Investigation on resistance of housefly in Zhangjiagang City. J. Med. Anim. Control 2007, 23, 112–113. (In Chinese) [Google Scholar]
- Li, F.; Chen, M. Efficacy of 9 insecticides against blattella germanica on passenger train. China J. Vector Biol. Control 2021, 12, 236. (In Chinese) [Google Scholar]
- Michael, E.S.; Walid, K. Changes in an insecticide-resistant field population of blattella germanica (Dictyoptera: Blattelidae) after expose to an insecticide mixture. Econ. Entomol. 1997, 90, 38–48. [Google Scholar]
- Valles, S.M.; Koehler, P.G. Comparative insecticide susceptibility and detoxification enzyme activities among pestiferous Blattodea. Comp. Biochem. Physiol. Part C 1999, 124, 227–232. [Google Scholar] [CrossRef]
- Gorun, V.; Proinov, L.; Baltescu, V. Modified Ellman procedure for assay of cholinesterases in crude enzymatic preparations. Anal. Chem. 1978, 86, 324–326. [Google Scholar] [CrossRef]
- Wu, X.F.; Deng, J.H. Effect of continuous use of pesticides on the development of resistance of Myzus persicae. China J. Tob. 2019, 10, 38–42. (In Chinese) [Google Scholar]
- Xia, S. Molecular Toxicology Basis; Wu, Z., Ed.; Hubei Science and Technology Press: Wuhan, China, 2000; pp. 66–67. (In Chinese) [Google Scholar]
- Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantites of protein utilizing the principle of protein dybinging. Anal. Chem. 1976, 72, 248–254. [Google Scholar]
- Ren, S.; Liu, Y.; Yang, J. Toxic effect of organophosphorus pesticide dimethoate on fish. J. Inn. Mong. Univ. Natl. (Nat. Sci. Ed.) 2020, 21, 659–661. (In Chinese) [Google Scholar]
- Zhai, D.S.; Zhao, Y.; Du, B.S. Acute toxicity test of paraquat in mice. West China J. Pharm. Sci. 2020, 22, 59–61. (In Chinese) [Google Scholar]
- Zhao, Z.; Huo, X. Insecticide resistance mechanism and control strategy of blattella germanica. Chin. J. Health Insectic. 2018, 14, 505–508. (In Chinese) [Google Scholar]
- Tang, Y.; Sun, B.X.; Zhang, L.M. Resistance test of blattella germanica to conventional insecticides in Changchun City. China Health Insectic. 2018, 14, 378–379. (In Chinese) [Google Scholar]
- Yu, X. Research progress of blattella germanica surveillance. China J. Vector Biol. Control 2018, 19, 593–594. (In Chinese) [Google Scholar]
- Ge, C.; Wang, L.; Zhang, W. Investigation on resistance of blattella germanica to five insecticides in Fushun City. China Health Insectic. 2017, 13, 459–460. (In Chinese) [Google Scholar]
- Yu, X.; Zeng, J.; Liu, J. Investigation on population density and infestation status of cockroaches in Neijiang City from 2017 to 2019. Chin. J. Health Insectic. 2021, 27, 340–342. (In Chinese) [Google Scholar]
- Li, L.; Hu, Y.; Yu, J. Surveillance of cockroach density and investigation of insecticide resistance of Blattella germanica in Sichuan Province. Zhonghua Health Insectic. Instrum. 2022, 28, 394–398. (In Chinese) [Google Scholar]
- Liu, Y.; Liu, H.; Leng, P. Laboratory efficacy of gel baits with four different active ingredients against Blattella germanica. Chin. J. Vector Biol. Control 2020, 31, 559–564. (In Chinese) [Google Scholar]
- Kobayashi, M.; Komatsu, N.; Ooi, H.K. Prevalence of Blatticola blattae (Thelastomatidae) in blattella germanicaes Blattella germanica in Japan. J. Vet. Med. Sci. 2021, 83, 174–179. [Google Scholar] [CrossRef] [PubMed]
- Su, S.; Ye, B. Modern Medical Entomology; Higher Education Press: Beijing, China, 1999; pp. 182–183. (In Chinese) [Google Scholar]
- Ma, Y.; Huo, X.; Ma, M. Inhibitory effect of beta-cypermethrin on acetylcholinesterase of blattella germanica. China J. Vector Biol. Control 2014, 15, 266–268. (In Chinese) [Google Scholar]
- Zhang, G. Insecticide resistance and its management. Anhui Agric. Sci. 2022, 30, 512–514. (In Chinese) [Google Scholar]
- Hou, W.; Xin, J.; Lu, H. Resistance development characteristics of reared blattella germanica (Blattodea:Blattellidae) to chlorpyrifos. Sci. Rep. 2021, 11, 3505. (In Chinese) [Google Scholar] [CrossRef]
- Li, T.; Wu, Y.; Liu, Q. Analysis of monitoring results of insecticide resistance of Blattella germanica in Zhejiang Province in 2018. Chin. J. Vector Biol. Control 2022, 33, 462–465. (In Chinese) [Google Scholar]
- Zhang, C.; Zhang, J. Investigation on resistance of Blattella germanica to commonly used insecticides in Pingdingshan City from 2018 to 2020. China Health Insectic. Instrum. 2022, 28, 116–118. (In Chinese) [Google Scholar]
- Lu, C.; Zhang, J.; Zhao, J. Study on the change of resistance of Blattella germanica in Yangpu District of Shanghai City from 2014 to 2020. Shanghai J. Prev. Med. 2022, 34, 123–125. (In Chinese) [Google Scholar]
- Chao, G.; Lv, W.; Lu, B. Cockroach surveillance and resistance analysis of Blattella germanica in Xianyang City in 2018. Chin. J. Health Insectic. 2021, 27, 121–123. (In Chinese) [Google Scholar]
- Oppenoorth, F.J. Biochemistry and genetics of insecticide resistance. In Comprehensive Insect Physiology. Biochemistry and Pharmacology; Kerkut, G.A., Gibert, I., Eds.; Pergamon, Press: New York, NY, USA, 1985; Volume 12, pp. 731–773. [Google Scholar]
- Fournier, D.; Mutero, A. A modification of acetylcholinesterase as a mechanism of resistance to insecticides. Comp. Biochem. Physiol. 1994, 108c, 91–131. [Google Scholar] [CrossRef]
- Huo, X.; Yu, G.; Ma, Y. Induction of glutathione peroxidase by cypermethrin in blattella germanica. China Public Health 2021, 20, 1318–1319. (In Chinese) [Google Scholar]
- Cochran, D.G. Monitoring for Insecticide Resistance in Filed-Collected Strains of German Cochroach (Dictyoptera:Blattellidate). Econ. Entomol. 1989, 82, 336–341. [Google Scholar] [CrossRef] [PubMed]
- Zhou, H.; Ma, Y.; Chen, Z. Resistance surveillance of blattella germanica to 5 insecticides in Wuxi city. Jiangsu Prev. Med. 2021, 12, 51–52. (In Chinese) [Google Scholar]
Resistant Strain | Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | ||
---|---|---|---|---|
LD50 (µg/Insect) | Resistance Factor (R) | LD50 (µg/Insect) | Resistance Factor (R) | |
Sensitive | 0.0097 | 1.0000 | 0.0338 | 1.000 |
First Generation | 0.0105 | 1.0825 | 0.0500 | 1.4793 |
Second Generation | 0.0112 | 1.1546 | 0.0625 | 1.8491 |
Three Generations | 0.0124 | 1.2784 | 0.0719 | 2.1272 |
Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | ||||||
---|---|---|---|---|---|---|---|
Resistant Strain | Resistance Factor (R) | Insect Number | AchE Activity nmol/(mg·min) | Resistant Strain | Resistance Factor (R) | Insect Number | AchE Activity nmol/(mg·min) |
Sensitive (0) | 1.0000 | 10 | 80.94 ± 1.68 | Sensitive (0) | 1.0000 | 10 | 80.94 ± 1.68 |
First Generation (1) | 1.0825 | 10 | 75.36 ± 4.01 | First Generation (1) | 1.4793 | 10 | 85.48 ± 2.26 |
Second Generation (2) | 1.1546 | 10 | 83.02 ± 3.32 | Second Generation (2) | 1.8491 | 10 | 91.21 ± 2.87 |
Third Generations (3) | 1.2784 | 10 | 87.71 ± 1.85 | Third Generation (3) | 2.1272 | 10 | 99.37 ± 2.29 |
Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | |||||
---|---|---|---|---|---|---|
Comparison between Groups | Lower 95% Confidence Interval | Mean Difference | Upper 95% Confidence Interval | Lower 95% Confidence Interval | Mean Difference | Upper 95% Confidence Interval |
(1)–(0) | −8.7395 | −5.5750 * | −2.4105 | 1.9998 | 4.5380 * | 7.0762 |
(2)–(0) | −1.0835 | 2.0810 | 5.2455 | 7.7378 | 10.2760 * | 12.8142 |
(3)–(0) | 3.6075 | 6.7720 * | 9.9365 | 15.8898 | 18.4280 * | 20.9662 |
Model Name | Regression Equation | F Value | p Value | R2 Value |
---|---|---|---|---|
simple linear | y = 15.751151x + 63.811891 | 30.50020 | 0.0313 * | 0.93846 |
logarithmic function | y = 22.601107ln(x) + 79.286586 | 14.67241 | 0.0619 | 0.88004 |
quadratic function | y = 90.365633 − 21.184139x + 11.873487x2 | 144.63696 | 0.0587 | 0.99655 |
Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | ||||||
---|---|---|---|---|---|---|---|
Resistant Strain | Resistance Factor (R) | Insect Number | GST Activity mOD/(mg·min) | Resistant Strain | Resistance Factor (R) | Insect Number | GST Activity mOD/(mg·min) |
Sensitive (0) | 1.0000 | 10 | 0.74 ± 0.01 | Sensitive (0) | 1.0000 | 10 | 0.74 ± 0.01 |
First Generation (1) | 1.0825 | 10 | 0.58 ± 0.02 | First Generation (1) | 1.4793 | 10 | 0.82 ± 0.02 |
Second Generation (2) | 1.1546 | 10 | 0.76 ± 0.03 | Second Generation (2) | 1.8491 | 10 | 0.88 ± 0.01 |
Third Generations (3) | 1.2784 | 10 | 0.82 ± 0.02 | Third Generations (3) | 2.1272 | 10 | 0.91 ± 0.02 |
Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | |||||
---|---|---|---|---|---|---|
Comparison between Groups | Lower 95% Confidence Interval | Mean Difference | Upper 95% Confidence Interval | Lower 95% Confidence Interval | Mean Difference | Upper 95% Confidence Interval |
(1)–(0) | −0.1867 | −0.1610 * | −0.1353 | 0.0635 | 0.0830 * | 0.1025 |
(2)–(0) | −0.0067 | 0.0190 | 0.0447 | 0.1165 | 0.1360 * | 0.1555 |
(3)–(0) | 0.0583 | 0.0840 * | 0.1097 | 0.1525 | 0.1720 * | 0.1915 |
Model Name | Regression Equation | F Value | p Value | R2 Value |
---|---|---|---|---|
simple linear | y = 0.152950x + 0.590485 | 322.48702 | 0.0031 * | 0.99384 |
logarithmic function | y = 0.226949ln(x) + 0.737452 | 587.26554 | 0.0017 * | 0.99661 |
quadratic function | y = 0.517671 + 0.254233x − 0.032559x2 | 397.10658 | 0.0355 * | 0.99874 |
Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | ||||||
---|---|---|---|---|---|---|---|
Resistant Strain | Resistance Factor (R) | Insect Number | P450-O Activity nOD/(mg·min) | Resistant Strain | Resistance Factor (R) | Insect Number | P450-O Activity nOD/(mg·min) |
Sensitive (0) | 1.0000 | 10 | 2.33 ± 0.07 | Sensitive (0) | 1.0000 | 10 | 2.33 ± 0.07 |
First Generation (1) | 1.0825 | 10 | 2.50 ± 0.15 | First Generation (1) | 1.4793 | 10 | 2.88 ± 0.19 |
Second Generation (2) | 1.1546 | 10 | 2.86 ± 0.15 | Second Generation (2) | 1.8491 | 10 | 3.50 ± 0.15 |
Third Generation (3) | 1.2784 | 10 | 3.06 ± 0.20 | Third Generation (3) | 2.1272 | 10 | 4.08 ± 0.23 |
Beta-Cypermethrin Nanoemulsion | Beta-Cypermethrin Emulsion | |||||
---|---|---|---|---|---|---|
Comparison between Groups | Lower 95% Confidence Interval | Mean Difference | Upper 95% Confidence Interval | Lower 95% Confidence Interval | Mean Difference | Upper 95% Confidence Interval |
(1)–(0) | 0.0141 | 0.1780 * | 0.3419 | 0.3706 | 0.5540 * | 0.7374 |
(2)–(0) | 0.3651 | 0.5290 * | 0.6929 | 0.9926 | 1.1760 * | 1.3594 |
(3)–(0) | 0.5681 | 0.7320 * | 0.8959 | 1.5766 | 1.7600 * | 1.9434 |
Model Name | Regression Equation | F Value | p Value | R2 Value |
---|---|---|---|---|
simple linear | y = 1.537104x + 0.715077 | 113.43873 | 0.0087 * | 0.98267 |
logarithmic function | y = 2.232321ln(x) + 2.213410 | 33.70863 | 0.0284 * | 0.94399 |
quadratic function | y = 2.097981 − 0.386465x + 0.618365x2 | 1,217,237.88239 | 0.0006 * | 1.00000 |
Resistance Factor (R) | AchE Activity nmol/(mg·min) | GST Activity mOD/(mg·min) | P450-O Demethylase nOD/(mg·min) |
---|---|---|---|
2.0 | 95.31 | 0.89 | 3.80 |
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. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Shen, Y.; Li, Q.; Fang, F.; Yang, C.; Dong, Y.; Li, X.; Luo, Z.; Shen, X. Comparative Study on the Resistance of Beta-Cypermethrin Nanoemulsion and Conventional Emulsion in Blattella germanica. Toxics 2023, 11, 834. https://doi.org/10.3390/toxics11100834
Shen Y, Li Q, Fang F, Yang C, Dong Y, Li X, Luo Z, Shen X. Comparative Study on the Resistance of Beta-Cypermethrin Nanoemulsion and Conventional Emulsion in Blattella germanica. Toxics. 2023; 11(10):834. https://doi.org/10.3390/toxics11100834
Chicago/Turabian StyleShen, Yan, Qiong Li, Fujin Fang, Chuanli Yang, Yu Dong, Xiaoqin Li, Zhizhi Luo, and Xiaobing Shen. 2023. "Comparative Study on the Resistance of Beta-Cypermethrin Nanoemulsion and Conventional Emulsion in Blattella germanica" Toxics 11, no. 10: 834. https://doi.org/10.3390/toxics11100834