Evaluation of insecticide resistance in different field populations of Cabbage aphid, Brevicornye brassicae  (Hemiptera: Aphididae) to selected organophosphate insecticides

doi:10.21203/rs.3.rs-4937997/v1

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Evaluation of insecticide resistance in different field populations of Cabbage aphid, Brevicornye brassicae (Hemiptera: Aphididae) to selected organophosphate insecticides

https://doi.org/10.21203/rs.3.rs-4937997/v1

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The Cabbage aphid, Brevicoryne brassicae is the most devastating pest of cruciferous crops, and management strategies mainly rely on insecticidal treatment. In this study, we evaluated the susceptibility of different field populations of B. brassicae collected from eight major cruciferous crop-producing districts of Kashmir, India, in 2023. The Toxicity of Chlorpyrifos (25% EC Kohiban), Dimethoate (30% EC Tafgor), and Quinalphos (25% EC Flash) was evaluated by foliage spray method under laboratory conditions. Different populations showed varied levels of susceptibility to the insecticides evaluated at various concentrations. The variability of the LC₅₀ values was 3.71 mlL^− 1 for dimethoate, 4.61 mlL^− 1 for chlorpyrifos, and 3.97 mlL^− 1 for quinalphos. After comparing insecticides resistance ratios (RR) with most susceptible field populations, resistance ratio to quinalphos was highest in the range of 1.13-3.07-fold, followed by 1.10-2.17-folds for dimethoate and 1.07-2.9-fold for Chlorpyrifos. The highest level of resistance to dimethoate was found in Budgam (LC₅₀ = 7.32 ml ^L−1) and lowest in Anantnag (LC₅₀ = 2.52 ml L ^− 1). Resistance against quinalphos was highest in Baramulla (LC₅₀ = 8.45 mlL^− 1) and lowest in Anantnag (LC₅₀ = 2.75 mlL^− 1). When exposed to the same insecticide, Bandipora exhibits the lowest resistance (LC₅₀ = 3.18 mlL^− 1) and Anantnag the highest resistance (LC₅₀ = 6.92 mlL^− 1). Rotation of insecticides with different modes of action where populations have no, very low, or low levels of resistance could be helpful in the management of B. brassicae infestation.

Foliage spray

B. brassicae

Sensitivity of insecticide

Mortality

Resistance ratio

Lethal dose

Cabbage aphid, B. brassicae, is a significant pest affecting kale (Brassica oleracea var. acephala), an important vegetable cultivated globally for its high nutritional and economic value (Mutua et al 2024). As a cosmopolitan species, Europe, Asia-Pacific, North America, Africa, Australia, and New Zealand are among the regions where B. brassicae is found. Additionally, it can be found widely in both temperate and warm temperate regions of the globe (Carvalho et al., 2002, Carter and Sorensen, 2013; Raychaudhuri, 1980; Blackman and Eastop, 2000). With the exception of the Far North, these species are likewise widely distributed throughout the former US Europe, Asia-Pacific, North America, Africa, Australia, and New Zealand are among the regions where B. brassicae is found. Additionally, it can be found widely in both temperate and warm temperate regions of the globe (Carvalho et al., 2002, Carter and Sorensen, 2013; Raychaudhuri, 1980; Blackman and Eastop, 2000). With the exception of the Far North, these species are likewise widely distributed throughout the former USSR. SR. Many Indian states where cruciferous crops are grown have reported cases of it, including Uttarakhand, Punjab, West Bengal, Karnataka (Basavaraju et al., 1995), Himachal Pradesh (Tandon et al., 1977), Meghalaya (Sachan and Gangwar, 1990), Manipur (Raychaudhuri, 1978), Andhra Pradesh, Tamil Nadu, Mahastra, Gujarat, Uttar Pradesh (Singh et al., 1999), Jammu and Kashmir, Manipur, Tripura, and other states. (Ghosh et al., 1980). Compared to other aphids, B. brassicae prefers a colder temperature (Ghosh et al., 1980). It is most common on these crops from January to March, with the highest number recorded in March. This pest causes leaves to curl and turn yellow because adults and nymphs drain plant fluids. Aphids release a honeydew sticky substance, on plant foliage and increases the extension of sooty mold, which prevents photosynthesis from occurring (Anwar and Shafique (1999). Under extreme infestation conditions, reduces crop production by as much as 80% (Atwal, 1976; Khattak and Hamed, 1993). B. brassicae transmits numerous plant viruses in addition direct mechanical harm (Sobhy et al. 2020). Several techniques are employed to manage aphids, one of which is the application of pesticides. Since 1990s, it has been advised to manage aphids with organophosphates, carbamates, and neonicotinoids (DOA, 2009; DOA, 2015). Aphids are challenging pest to control due to several factors, including their reproduction, range of hosts, and the emergence of insecticide resistance. Due to insecticide resistance to a variety of substances, such as pyrethroids, carbamates, neonicotinoids, and organophosphates, most insecticides are currently inefficient worldwide (Fuentes-Contreras et al., 2013; Bass et al., 2014; Umina et al., 2014).

In India, chlorpyrifos has been registered since 1977 under the Insecticides Act. Furthermore, the Act permits the use of seven formulations of chlorpyrifos. The CIB & RC (Central Insecticide Board and Registration Committe) which is part of the PPQS (Ministry of Agriculture & Farmers Welfare, the Directorate of Plant Protection, Quarantine & Storage) and the GOI (Government of India) states that as of 2016–17, chlorpyrifos is the most widely used pesticide in India, accounting for 9.4% of all insecticide consumption. American Cyanamid patented and introduced dimethoate, an organophosphate pesticide, and acaricide, to India in the 1950s. Dimethoate inhibits acetylcholinesterase, just like other organophosphates do, impairing cholinesterase activity (Dauterman et al., 1960). Organophosphate such as quinalphos is used as a broad-spectrum insecticide and acaricide to combat a variety of common insect pests, including mites, aphids, and mealy bugs (Srivastava et al., 2016; Deosi et al., 2022).

Resistance has been reported in many countries against these insecticides. Pesticide resistance is a worldwide problem and a heritable characteristic. There may be several underlying mechanisms for the development of resistance in insects. The two most popular methods are improved enzyme-assisted pesticide detoxification and target site mutation, which results in insensitive target sites (WHO, 1998. Although B. brassicae has been the target of numerous control strategies (Munthali and Tshegofatso 2014). In the Indian subcontinent, these compounds are still used to suppress sucking insects (Sethi et al., 2008) and it frequently needs repeated applications (Joshi et al. 2010). Understanding the level of resistance exhibited by a specific insect pest is essential for assessing the efficacy of current management approaches and formulating novel tactics.

Being a predominant pest of cruciferous crops, different test populations of B. brasssicae were collected from major cruciferous crop-producing areas where the use of these insecticides occurs frequently. As the number of spray schedules of these insecticides has been found to increase year after year, however, crop quality and production are not enough to compensate for farmer’s costs. To the best of our knowledge, monitoring of resistance against insecticides in B. brassicae has not yet been reported from cruciferous-producing areas growing in Kashmir, India. The novel study was, therefore, undertaken to evaluate the resistance status and its regional variation in this pest to three conventional and commonly used insecticides chlorpyrifos, dimethoate, and quinalphos. Alternative methods of managing agricultural pests, as well as more advanced and effective formulations, can be prepared with a deeper understanding of resistance distribution.

Insects

The adults of Brevicorynae brassicae were identified following the morphological characters and were collected from infested kale plants, Brassica oleracea var. acephala. To evaluate the insecticide susceptibility and status of resistance to test insecticides, eight populations of Brassica oleracea var. acephala were individually sampled in 2023 from the eight most significant provinces as shown in Fig. 1. The geographic coordinates of sampling locations from each district are shown in Table 1. These were treated as eight different populations based on location. Infested leaves of Brassica oleracea var. acephala were collected in the crop growing season. Infested, as well as healthy leaves were individually collected in plastic bags and transferred to the Entomological laboratory after collection; they were treated directly. Adult apterous aphids were used for the study.

Table 1

Collection sites of different field populations of cabbage aphid, *B. brassiacae* in Kashmir
Region	Study location	Sampling site	Latitude	longitude
South Kashmir	Anantnag	Banghidar	33.7294^º	75.1446^º
	Pulwama	Kati Bugh	33.9000 ^º	74.9057^º
	Kulgam	Wanpoh	33.7255 ^º	75.1039^º
Central Kashmir	Budgam	Bugam Batpora	33.9545 ^º	74.8414^º
	Srinagar	Shanglipora	34.0551^º	74.4645^º
	Ganderbal	Zazna	34.2256^º	74.6868 ^º
North Kashmir	Bandipora	Plan Bandipora	34.4138^º	74.6382^º
North Kashmir	Baramulla	Stadium colony	34.2119^º	74.3510^º

Insecticides

Chlorpyrifos (Kohiban @ 25%, EC; FIL Industries Private Limited, Kashmir, India), Dimethoate (Tafgor @ 20%, EC; TATA Rallis, Maharashtra, India), Quinalphos (Flash @ 30%, EC; Indofill Industries Limited, Gujarat, India) were obtained from their respective manufacturers. These selected insecticides represent organophosphates being commonly used in the fields where the cruciferous crop is produced for commercial purposes to control B. brassicae. Moreover, these compound labels claimed their efficacy against B. brassicae by the insecticides registered use with the CIBRC (Central Insecticides Board and Registration Committee, Government of India) as of the relevant date.

Preparation of stock solution

For toxicity tests, different concentrations of each insecticide viz dimethoate, chlorpyrifos and quinalphos were prepared from their respective stock solutions. The solutions of 0.25 ml, 0.50ml, 0.75ml, 1ml and 1.5ml per liter of water were thoroughly mixed with a glass rod for proper dilution. After the preparation of the solutions, the cap of each flask was covered with parafilm to minimize evaporation and were kept in the freezer (preferably − 20°C) or in refrigerator (4°C) for further use.

Bioassay

The foliar spray method was used to conduct bioassays against three insecticides (Patil et al., 2017). Leaf discs of 5–10 cm were prepared from a selected cruciferous plant. Leaf discs (5cm) were placed in a Petri dish and directly sprayed with different concentrations viz 0.25ml, 0.50 ml, 0.75 ml,1ml, and 1.5ml of test insecticides using Potter`s tower at a particular pressure (Heron et al., 1993, Patil et al., 2017). The adaxial side of the treated leaf discs was positioned upright in plastic Petri plates and a moist filter paper was positioned underneath. The sprayed leaf discs were dried for five minutes at room temperature. Finally, 20 apterous adults from B. brassicae var. acephala were transferred on dried leaf disc with the help of camel hair brush. This procedure was repeated for each concentration with three replicates. After that, the Petri dishes were enclosed with muslin cloth and kept in a controlled conditions with a temperature of 25 ± 2°C and a relative humidity of 50 ± 10%. The light-to-darkness ratio was set at 14 hours to 10 hours. The same number of adults as a check (Control) was treated with distilled water, and air-dried.

Data analysis

Mortality counts were taken 24 hours after exposure. Adult insects displaying poor reaction to external stimuli or lack of coordinated movement when gently touched with a fine camel hairbrush was noticed. Mortality between control and treatment was corrected by using Abbot's formula (Abbot 1925. Aly et al., 2013). R analysis was performed using the R software (version 4.2.3)) to determine the lethal concentration (LC₅₀ and LC₉₀) Lethal 95% fluidical limits, χ2 significance tests, standard error and slope of the regression lines (Munir et al., 2005, Ishtiaq et al., 2012)

Percent mortality (%) =\(\:\:\:\:\frac{\:\:\:\:\text{N}\text{o}.\:\text{o}\text{f}\:\text{i}\text{n}\text{d}\text{i}\text{v}\text{i}\text{d}\text{u}\text{a}\text{l}\text{s}\:\text{i}\text{n}\:\text{t}\text{r}\text{e}\text{a}\text{t}\text{m}\text{e}\text{n}\text{t}\:\left(\text{P}\text{T}\right)\:\:}{\:\:\:\:\text{N}\text{o}.\:\text{o}\text{f}\:\text{i}\text{n}\text{d}\text{i}\text{v}\text{i}\text{d}\text{u}\text{a}\text{l}\text{s}\:\text{i}\text{n}\:\text{c}\text{o}\text{n}\text{t}\text{r}\text{o}\text{l}\:\left(\text{P}\text{C}\right)\:\:}\:\)x 100

The population with the lowest LC50 or LC90 was used as a reference, and the resistance ratio of each insecticide among different populations was calculated (Hounded et al., 2010; Shadmany et al., 2015) by dividing the LC50 of the test population with LC50 of susceptible population.

Resistance ratio|(RR)=\(\:\:\:\:\frac{\:\:\:\:\text{L}\text{e}\text{t}\text{h}\text{a}\text{l}\:\text{c}\text{o}\text{n}\text{c}\text{e}\text{n}\text{t}\text{r}\text{a}\text{t}\text{i}\text{o}\text{n}\:\left(\text{L}\text{C}50\right)\:\text{l}\text{e}\text{t}\text{h}\text{a}\text{l}\:\text{o}\text{f}\:\text{t}\text{h}\text{e}\:\text{t}\text{e}\text{s}\text{t}\:\text{p}\text{o}\text{p}\text{u}\text{l}\text{a}\text{t}\text{i}\text{o}\text{n}}{\:\:\:\:\:\text{L}\text{e}\text{t}\text{h}\text{a}\text{l}\:\text{c}\text{o}\text{n}\text{c}\text{e}\text{n}\text{t}\text{r}\text{a}\text{t}\text{i}\text{o}\text{n}\:\left(\text{L}\text{C}50\right)\text{l}\text{e}\text{t}\text{h}\text{a}\text{l}\:\text{o}\text{f}\:\text{t}\text{h}\text{e}\:\text{s}\text{u}\text{s}\text{c}\text{e}\text{p}\text{t}\text{i}\text{b}\text{l}\text{e}\:\text{p}\text{o}\text{p}\text{u}\text{l}\text{a}\text{t}\text{i}\text{o}\text{n}\:}\:\)

Regression analysis was employed to assess the significance of pairwise comparisons between test insecticide concentrations and mortality rates. Correlation analysis was performed using R Studio software (version 4.2.3). The 95% confidence limits and the estimated LC₅₀ and LC₉₀ to the maximum recommended field dose established by the (CIBRC) was compared. In India, the advised label dose for test insecticides against sucking pests range from 0.25 ml/L to 0.75 ml/L. The mortality attained at the label rate would be regarded as notably lower than the complying label if the LC50 at 95% fiducial limits was discovered to be greater than the advised rate.

Concentration-mortality pairwise correlation analysis of three insecticides against eight populations of B. brassicae.

Percent mortality at different concentrations of the tested insecticides against B. brassicae field-collected populations was assessed by parson correlation analysis as shown in Fig. 2. Upon testing varying dosages of dimethoate, Chlorpyriphos, and quinalphos (0.25 ml, 0.50 ml, 0.75 ml, 1.0 ml, and 1.5 ml) against distinct populations of cabbage aphids, B. brassicae variable range of mortality was found. Treatment with dimethoate showed a positive correlation between insecticide concentration and mortality in Anantnag populations ((Pearson’s correlation coefficient R² = 0.84, P < 0.001), while treatment with chlorpyrifos shows weakly positive correlation (R² = 0.51, P < 0.001). When aphid populations from Bandipora were treated with quinalphos, mortality was found significantly correlated (R² = 0.86, P < 0.001), while treating them with dimethoate showed a weakly positive correlation (R² = 0.58, P < 0.001). In contrast, the correlation between insecticide concentration and mortality in Baramulla population was found to be lowest when treated with quinalphos, mortality was found significantly correlated (R2 = 0.86, P < 0.001), while treating them with dimethoate showed a weakly positive correlation (R² = 0.58, P < 0.001). In contrast, the correlation between insecticide concentration and mortality in Baramulla population was found to be lowest when treated with quinalphos concentrations (R² = 0.40, P = 0.005) and positively correlated when treated with dimethoate (R² = 0.84, P < 0.001). When Chlorpyriphos was used, mortality was shown to be positively correlated (R² = 85, P < 0.001), whereas, at greater doses, dimethoate showed saturation (R² = 0.70, P < 0.001) in the Budgam population. In the case of Ganderbal populations, mortality was found weakly correlated when treated with chlorpyrifos (R² = 0.82, P < 0.001) and highly positively correlated when treated with dimethoate (R² = 0.83, p < 0.001). Population mortality in Kulgam was found to be low when treated with quinalphos showing saturation at lower concentrations (R² = 0.77, P = 0.005) and saturation was found after 30% mortality when treated with increasing concentrations of dimethoate (R² = 0.80, P = 0.001). Mortality was found positively correlated with insecticide concentration of Dimethoate and Chlorpyrifos (R² = 0.89, P < 0.001, R² = 0.83, P < 0.001) in Pulwama populations. When quinalphos was administered to the Srinagar population, mortality showed a sigmoid pattern of mortality (R² = 0.84, p < 0.001), but dimethoate treatment resulted in an almost linear trend in mortality (R² = 0.82, P < 0.001). Our findings are consistent with the work of Zhang et al., 2022, which assessed insecticide resistance in field-collected populations of S. litura sampled from eleven Chinese provinces in 2022.

Baseline Susceptibility of B. brassicae populations to three Insecticides

Table 2. summarizes the results of the probit analysis's dose mortality response for each insecticide that was tested. The results indicate chlorpyrifos to have LD₅₀ and LD₉₀ values of 4.6 mlL^− 1 (3.24 ± 7.86) and 78.77 mlL^− 1 (33.54 ± 300.22) followed by quinalphos with LD₅₀ and LD₉₀ of 3.97 mlL^− 1 (2.89 ± 6.43) and dimethoate with LD₅₀ and LD₉₀ of 3.71 mlL^− 1 (2.77 ± 5.69) and 50.68 mlL^− 1 (24.65 ± 150.15). Recorded slope values for chlorpyrifos, quinalphos and dimethoate were 1.04 ± 0.12, 1.04 ± 0.15 and 1.13 ± 0.12, respectively.

Table 2

Effect of insecticides against adult *B. brassicae*, slope, chi-square, P. value, LC₅₀, and LC₉₀ of Chlorpyrifos, dimethoate, and quinalphos after 24 hours of treatment.
Insecticide	N	Slope	chi-square	df	P	LC₅₀ (95%FL)	LC₉₀ (95%FL)
Quinalphos	120	1.04 ± 0.12	65.65	118	0.99997	3.97(2.89 ± 6.43)	67.16(29.92 ± 234.65)
Chlorpyriphos	120	1.04 ± 0.12	57.77	118	0.99999	4.61(3.24 ± 7.86)	78.77(33.54 ± 300.22)
Dimethoate	120	1.13 ± 0.12	33.55	118	0.99999	3.71(2.77 ± 5.69)	50.68(24.65 ± 150.15)
N- Number of individuals tested; df-degree of freedom; P - Significance value; FL- fiducial limit

Lethal concentrations of Chlorpyriphos, Dimethoate and Quinalphos.

The LC₅₀ and LC₉₀ values were calculated by r analysis using R studio. Abbott (1925) formula of corrected mortality were used to find out mortality rates. LC₅₀ concentration of quinalphos was found approximately 3.97mlL^− 1 and LC₉₀ concentration was approximately around 68 ml. Chlorpyriphos requires the highest LC₅₀ concentration, with over 4.61 ml^− 1, and an LC₉₀ concentration of nearly 78 mlL^− 1. In the case of dimethoate LC₅₀ concentration was around 3.71 mlL^− 1 ml and LC₉₀ concentration was significantly less 50.68 mlL^− 1 as shown in Fig. 3.

Dose-effect curves of dimethoate, Chlorpyriphos and quinalphos against B. brassicae populations

Dose-effect mortality curves shows percent mortality of Chlorpyriphos (blue line) starts at a lower concentration but increases steadily as the concentration rises. Dimethoate (red line) begins at a similar point as Chlorpyrifos but has a steeper increase in percent mortality with increasing concentrations. Quinalphos (green line) also starts at low concentration, but its increase is more gradual compared to dimethoate. Overall, there is a positive correlation between insecticide concentration and effectiveness in causing mortality. As the concentration of each insecticide increases, the % mortality also increases. Dimethoate shows the most pronounced effect on mortality, followed by chlorpyrifos and then quinalphos as shown in Fig. 4.

Regional variation in lethal concentrations (LC ₅₀ ) of dimethoate insecticide against different populations of Brevicorynae brassicae.

The highest LC₅₀ value 7.32 mlL^− 1 was found in the case of Budgam populations, followed by a decreasing trend from Budgam (7.32 mlL^− 1) to Anantnag (2.52 mlL^− 1) populations. However, an abrupt decrease in LC₅₀ values was found in Pulwama populations measuring 3.57 mlL^− 1, and the lowest LC₅₀ was found in Anantnag populations measuring 2.52 mlL^− 1. The results have shown the Budgam population most resistant to dimethoate and the Anantnag population to be most susceptible as shown in Fig. 5.

Variation in lethal concentration (LC ₅₀ ) of Chlorpyrifos insecticide against different populations of B. brassicae.

In case of chlorpyrifos, the highest LC₅₀ value measuring 6.92 mlL^− 1 was found in the case of Anantnag populations, falling lowed by a gradual decrease in LC₅₀ values from Anantnag with 6.92 ml^− 1to Bandipora 3.18 mlL^− 1. However, the LC₅₀ value was found similar in case of the Budgam and Kulgam populations measuring 3.53 mlL^− 1 respectively. The results have shown Anantnag population was most resistant to chlorpyrifos and the Bandipora population was found most susceptible to test insecticide as shown in Fig. 6.

Regional variation lethal concentration (LC ₅₀ ) of quinalphos insecticide against different populations of B. brassicae.

The highest LC₅₀ value measuring 8.45 mlL^− 1 was found in the case of Baramulla populations followed by a gradual decrease in trend from Ganderbal (5.57 mlL^− 1) when treated with Chlorpyrifos to Srinagar (3.17 mlL^− 1) populations. However abrupt decrease in LC₅₀ values was found in Anantnag populations measuring 2.75 mlL^− 1. The results have shown the Baramulla population as most resistant and the Anantnag population is most susceptible when treated with quinalphos as shown in Fig. 7.

Sensitivity of three Insecticides in different populations of B. brassicae

Resistance ratios (RR) also have shown variation among the eight field-collected populations of B. brassicae from Kashmir. Anantnag population (RR = 1) was found most susceptible when treated with dimethoate. Pulwama (RR = 1.41), Baramulla (RR = 1.24), Kulgam (RR = 1.07), and Ganderbal (RR = 1.08) populations showed almost similar levels of resistance. Bandipora (RR = 2.35) and Srinagar (RR = 2.64) showed a moderate level of resistance, Highest level of resistance against dimethoate was found in the Budgam population (RR = 2.90) as shown in Table 3. The tested populations displayed low to moderate levels of resistance against Chlorpyriphos. Likewise, Budgam (RR = 1.11), Pulwama (RR = 1.62), Kulgam (RR = 1.10), and Srinagar ((RR = 1.26) populations displayed the same level of resistance to chlorpyrifos. High resistance level against Chlorpyriphos was found in the Ganderbal population (RR = 2.02), Baramulla (RR = 2.13), and Anantnag (2.17) as you compared to the most susceptible Bandipora population (RR = 1) as shown in Table 4. The field-collected populations from Budgam (RR = 1.17), Pulwama (RR = 1.95), Kulgam (RR = 1.23), Srinagar (RR = 1.15), and Bandipora (RR = 1.13) showed lowest level of resistance to quinalphos. Baramulla (RR = 3.07) and Ganderbal (RR = 2.02) populations displayed higher resistance level to quinalphos as compared to the most susceptible population Anantnag (RR = 1.1) as shown in Table 5.

Table 3

Insecticide (dimethoate) susceptibility of *B, brassiacae* collected from different location of Kashmir.
Site	N	Slope	chi-square	df	p	LC₅₀	RR
Baramulla	15	1.28228	1.59699	13	0.99994	3.13001	1.24176
Budgam	15	0.8712	3.39925	13	0.99607	7.3206	2.90429
Pulwama	15	1.15154	1.38044	13	0.99997	3.57199	1.41711
Kulgam	15	1.13771	1.68763	13	0.99991	2.69983	1.0711
Srinagar	15	0.84294	1.16516	13	0.99999	6.67911	2.64979
Anantnag*	15	1.29009	2.37147	13	0.99942	2.52062	1
Bandipora	15	0.9908	7.95306	13	0.84664	5.92345	2.35
Ganderbal	15	1.5174	3.38217	13	0.99617	2.74344	1.0884
*: Most Susceptible. N: Number of individuals tested. df: degree of freedom. P: significant value. RR: Resistance Ratio = LC50 (field-collected population)/LC₅₀ (most Susceptible (S)

Table 4

Insecticide (chlorpyrifos) susceptibility of *B, brassiacae* collected from different location of Kashmir.
Site	N	slope	chi-square	df	p	LC₅₀	RR
Baramulla	15	0.7527	14.63189	13	0.33089	6.781	2.13151
Budgam	15	1.3777	2.52684	13	0.99917	3.53381	1.1108
Pulwama	15	1.11308	1.9718	13	0.99979	5.15421	1.62015
Kulgam	15	0.9888	2.02809	13	0.99976	3.52535	1.10814
Srinagar	15	1.13417	1.97153	13	0.99979	4.03241	1.26753
Anantnag	15	0.90672	9.57914	13	0.72793	6.92045	2.17535
Bandipora*	15	1.30762	5.0675	13	0.97368	3.18131	1
Ganderbal	15	0.86901	6.81247	13	0.91156	6.45642	2.02949
*: Most Susceptible. N: Number of individuals tested. df: degree of freedom. P: significant value. RR: Resistance Ratio = LC₅₀ (field-collected population)/LC₅₀ (most Susceptible).

Table 5

Insecticide (quinalphos) susceptibility of *B, brassiacae* collected from different location of Kashmir.
Site	N	slope	chi-square	df	p	LC₅₀	RR
Baramulla	15	0.50927	12.9064	13	0.45507	8.44862	3.07402
Budgam	15	1.19589	3.44459	13	0.99579	3.2269	1.1741
Pulwama	15	1.04141	3.25863	13	0.99683	5.36862	1.95337
Kulgam	15	1.20865	5.17753	13	0.97107	3.40166	1.23769
Srinagar	15	1.25042	4.19575	13	0.98893	3.16725	1.1524
Anantnag^S	15	1.30577	10.08295	13	0.68714	2.7484	1.0000
Bandipora	15	1.06725	0.82213	13	1	3.82606	1.39211
Ganderbal	15	0.9623	6.47152	13	0.9273	5.56766	2.02578
*: Most Susceptible. N: Number of individuals tested. df: degree of freedom. P: significant value. RR: Resistance Ratio = LC₅₀ (field-collected population)/LC₅₀ (field collected most Susceptible).

Insecticide resistance monitoring is a critical component of resistance management (Sparks et al., 2020). In the current study upon testing varying dosages of dimethoate, chlorpyriphos, and quinalphos against different populations of cabbage aphid, B. brassicae, a wide range of mortality was observed. Dimethoate is marginally more effective than Chlorpyrifos at lower concentrations. However, in Budgam, Srinagar, and Bandipora populations, dimethoate has the steepest slope and shows a mortality rate lower than the other two. The LC₅₀ of dimethoate against these populations have shown resistance ratio (RR) of 2.9, 2.6, and 2.3-fold higher than the field susceptible population. Resistance against dimethoate has also been found in Cotton seed bug, Oxycarenus hyalinipennis where some populations have developed high level of resistance (Banazeer et al., 2020). In Anantnag, Baramulla, and Pulwama populations the concentration of all three insecticides shows variable correlation between mortality rate and insecticide concentration. Chlorpyrifos, which has the steepest slope, shows a mortality rate lower than the other two. The LC₅₀ of chlorpyrifos against these populations was found 2.17, 2.13, and 1.6-fold higher than the field susceptible population. Research has also shown that numerous insects, including Bemisia tabaci (Zhang et al., 2012), Phenacoccus solenopsis (Afzal et al., 2015), Drosophila melanogaster (Daisley et al., 2018), and Spodoptera littoralis (Ismail, 2020), have developed resistance to chlorpyrifos pesticides. The LC50 of Baramulla, Ganderbal, and Pulwama against quinalphos was 2.9, 2.0, and 1.9.3-fold higher than the field susceptible population. The results revealed that above tested organophosphates have shown low to moderate levels of resistance. Additionally, it has been reported that the oriental fruit fly, Bactrocera dorsalis to trichlorphon (Yu-ping et al., 2010), cotton mealybug, Phenacoccus solenopsis to chlorpyrifos (Afzal et al., 2015), cotton leafworm, S. litura to profenofos (Abbas et al., 2014), and southern house mosquito, Culex quinquefasciatus to dimethoate (Alam et al., 2017), are also reported to be resistant to insecticides of the organophosphate group. Removing that specific insecticide from the spray schedule and vice versa may diminish resistance in certain places, as indicated by the decline in the resistance ratio in some populations (Ejaz et al., 2017). However, no insecticide was found fully toxic against all tested populations. The cause of high susceptibility against a particular insecticide in some field populations could be related to indiscriminate use of that particular insecticide in these areas and vice versa (Ishtiaq et al., 2012). Base susceptibility analysis has shown higher Chlorpyrifos resistance with LC50 and LC90 of 4.61 and 78.77 than other tested insecticides. Very high resistance to Chlorpyrifos has been documented in provinces of Mexico and Guatemala (Delorme et al., 1988, Teran-Vargas et al., 1999). In Punjab, Pakistan, resistance to chlorpyrifos was found in the cotton leafworm S. littoralis (Ahmad et al., 2007a, b; Saleem et al., 2008), suggesting different level of resistance variation in resistance to organophosphates. Similar variability was seen in tested populations, with some field populations showing higher tolerance to multiple insecticides compared to the reference susceptible strain (Toscano et al., 2001). Additionally, correlation analysis also revealed cross-resistance pattern in B. brassicae to all tested insecticides (Zhang et al., 2022). Variations in resistance levels to the insecticide triazophos have been observed in various populations of B. tabaci from Turkey, Pakistan, and India. The study's findings about the emergence of chlorpyrifos resistance are consistent with reports of resistance in B.tabaci from Pakistan. While the current level of resistance is not immediately alarming and is unlikely to significantly affect field efficacy, it does indicate a potential risk of increasing resistance, especially if the cabbage aphid, B. brassicae, becomes too reliant on these insecticides (Hopkinson et al., 2023).

Studies on insecticide resistance can be utilized to create evidence-based insecticide control programs for B. brassicae, reducing the excessive insecticides use. Evaluation of B.brassicae resistance against synthetic insecticides in other crops will improve our understanding of management strategies for this pest. However, major issue is that most of the farmers untrained and illiterate, resulting in frequent errors when insecticides are applied to crops. Formers often apply incorrect doses of insecticide for specific pests and are usually unaware of the EILs (Economic Injury Levels) and ETLs (Economic Threshold Levels) of pests. The indiscriminate application of pesticides at regular intervals results from lack of knowledge. It's possible that this may be also the reason why pests gain resistance. The growing resistance problem requires the development of region-specific strategies to address resistance issues for nearly all pests.

Cabbage aphid, a substantial pest of cruciferous crops, that are very important vegetable crops grown worldwide for their high economic and value nutritional. Synthetic insecticides chlorpyrifos, dimethoate, and quinalphos are the most utilized to manage this pest, often requiring repeated applications. The current study of insecticide resistance to the above-tested insecticides in B. brassicae has not been yet reported in Kashmir, India to the best of our knowledge. The test insecticides had varying effects on different populations of B. brassicae. At lower concentrations, dimethoate showed a slight advantage over chlorpyrifos. Dimethoate had the steepest slope yet the lowest mortality rates in certain areas (Budgam, Srinagar, and Bandipora). Chlorpyrifos had a steeper slope but still had lower fatality rates in other regions (such as Anantnag, Baramulla, and Pulwama). Quinalphos efficacy was found inconsistent. All studied populations were not completely susceptible to any single tested insecticide. Optimally, non-chemical control approaches should be employed in conjunction with insecticide-based strategies like the insecticide-based "MoA (Mode of Action) treatment windows" approach suggested by IRAC (Insecticide Resistance Action Committee). This is part of a larger integrated pest management (IPM) strategy. Treating successive generations of the target pest with insecticides having the same MoAc (Mode of Action Classification) is not a recommended course of action. Using insecticides with a different MOA, any necessary further treatments after the specified treatment window should be carried out. The development of resistance to these pesticides may be slowed down by rotating the treatment schedule of organophosphate insecticides with insecticides of other classes. Growers should also carefully follow the product label regarding the quantity, procedure, target pest, and pace of insecticide treatment.

Conflict of interest

The authors have no competing interests either financially or non-financially.

Funding

No funding was obtained for this study.

Author Contribution

Tariq Ahmad and Barkat Hussain gave original concept, experimental design of this manuscript. Irfan Ahmad Mir carried out the experiments and wrote initial draft & data analysis. The final draft was reviewed and approved by all authors

Availability of Data

The present manuscript does not have any research data outside the submitted manuscript file

Abbas, N., Shad, S. A., Razaq, M., Waheed, A., & Aslam, M. (2014). Resistance of Spodoptera litura (Lepidoptera: Noctuidae) to profenofos: Relative fitness and cross resistance. Crop Protection, 58, 49–54.
Abbott, S. W. (1925). A method of computing the effectiveness of an insecticide, Journal of Economic Entomology, 18, 265–267.
Afzal, M. B. S., Shad, S. A., & Abbas, N. (2015). Genetics, realized heritability and preliminary mechanism of spinosad resistance in Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae): An invasive pest from Pakistan. Genetica, 143, 741–749.
Afzal, M.B.S., M. Ijaz, Z. Farooq, S.A. Shad, and N. Abbas. 2015. Genetics and preliminary mechanism of chlorpyrifos resistance in Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae). Pesticide Biochemistry and Physiology 119: 42–47.
Ahmad, M. U. N. I. R., & Aslam, M. U. H. A. M. M. A. D. (2005). Resistance of cabbage aphid, Brevicoryne brassicae (Linnaeus) to endosulfan, organophosphates and synthetic pyrethroids. Pakistan Journal of Zoology, 37(4), 293.
Alam, M., Sumra, M. W., Ahmad, D., Shah, R. M., Binyameen, M., & Shad, S. A. (2017). Selection, realized heritability, and fitness cost associated with dimethoate resistance in a field population of Culex quinquefasciatus (Diptera: Culicidae). Journal of Economic Entomology, 110, 1252–1258.
Aly, A. Ella, A. (2013). Toxicity and persistence of selected neonicotinoid insecticides on cowpea aphid, Aphis craccivora Koch (Homoptera: Aphididae), Archives of Phytopathology and Plant Protection, 3, 366–376.
Anwar, M. and Shafique, M., (1999). Relative development of aphids on different Brassica cultivars. Pakistan J. Zool., 31: 357–359.
Atwal, a.s., (1976). Agricultural pests of India and Southeast Asia, Kalyani Publishers, New Delhi, India, pp. 310–311.
Banazeer, A., Afzal, M. B. S., & Shad, S. A. (2020). Characterization of dimethoate resistance in Oxycarenus hyalinipennis (Costa): resistance selection, cross-resistance to three insecticides and mode of inheritance. Phytoparasitica, 48, 841–849.
Basanth Y S, Sanna veerappanavar V T and Gowda D K S. 2013. Susceptibility of different populations of Nilaparvata lugens from major rice growing areas of Karnataka, India to different groups of insecticides. Rice Science 20(5): 371–73.
Basavaraju, B.S., Rajagopal, B.K., Sherrif, R.A., Rajagopal, D. and Jagadish, K.S. (1995). Seasonal abundance of aphids on mustard Brassica juncea (L.) Czern and Coss at Bangalore. Mysore J. Agric. Sci., 29(3): 225–229.
Bass, C., & Nauen, R. (2023). The molecular mechanisms of insecticide resistance in aphid crop pests. Insect Biochemistry and Molecular Biology, 103937.
Bass, C., Puinean, A.M., Zimme, C., Denholm, I., Field, L. M., Foster, S.P., Gutbrod, O., Nauen, R., Slater, R., & Williamson, M.S., (2014). The evolution of insecticide resistance in the peach potato aphid, Myzus persicae. Insect Biochemistry and Molecular Biology, 51, 41–51.
Blackman, R.L. and Eastop, V.F. (2000). Aphids on the World’s Crops: an Identification Guide, 2nd ed. Wiley, New York.
Calculated using data from the PPQS Statistical Database. http://ppqs.gov.in/statistical-database.
Carter, C.C. and Sorensen, K.A. (2013). Insect and related pests of vegetables. Cabbage and turnip aphid. Center for Integrated Pest Management. North Carolina State University, Raleigh, NC.
Carvalho, L.M. de., Bueno, V.H.P. and Martinez, R.P. (2002). Alate aphids survey on vegetable crops in Lavras (MG). Cien.Agrotecnol. 26(3): 523–532.
Chen, B.K., Wang, K.Y., Jiang, X.Y., Yi, M.Q., 2002. Studies and surveys on the insecticide resistance of Spodoptera exigua. Acta Phytoph. Sin. 29, 366e370.
Daisley, B.A., M. Trinder, T.W. McDowell, S.L. Collins, M.W. Sumarah, and G. Reid. 2018. Microbiota-mediated modulation of organophosphate insecticide toxicity by species-dependent interactions with lactobacilli in a Drosophila melanogaster insect model. Applied and Environmental Microbiology 84(9): e02820–e2917.
Dauterman WC, Viado GB, Casida JE, O'brien RD (1960). "Insecticide Residues, Persistence of Dimethoate and Metabolites Following Foliar Application to Plants". Journal of Agricultural and Food Chemistry. 8 (2): 115–9.
Delorme, R., Fournier, D., Chaufaux, J., Cuany, A., Bride, J.M., Auge, D., Berge, J.B., 1988. Esterase metabolism and reduced penetration are causes of resistance to deltamethrin in Spodoptera exigua (Hubner) (Noctuidae: Lepidoptera). Pestic. Biochem. Physiol. 32, 240e246.
Deosi, K. K., & Suri, K. S. (2022). Status of insecticide resistance in rice brown planthopper (Nilaparvata lugens) in Punjab. Indian Journal of Agricultural Sciences, 92(2), 203–207.
Ejaz, M., Afzal, M. B. S., Shabbir, G., Serrão, J. E., Shad, S. A., & Muhammad, W. (2017). Laboratory selection of chlorpyrifos resistance in an Invasive Pest, Phenacoccus solenopsis (Homoptera: Pseudococcidae): Cross-resistance, stability and fitness cost. Pesticide biochemistry and physiology, 137, 8–14.
Fuentes-Contreras, E., Figueroa, C. C., Silva, A.X., Bacigalupe, L. D., Briones, L. M., Foster, S. P.& Unruh, R. (2013). Survey of Resistance to Four Insecticides and their Associated Mechanisms in Different Genotypes of the Green Peach Aphid (Hemiptera: Aphididae) From Chile. Journal of Economic Entomology,106(1), 400–407.
Ghosh, L.K., Raychaudhuri, D., Raychaudhuri D.N. and Raha, S.K. (1980). An account of the genus Brevicoryne Goot (Homoptera: Aphididae) in India with description of sexual morph of B. barbarae Nevsky. Bull. Zool. Surv. India, 3: 63–68.
Heron, A. G., Gibson, S. T. and M. A. Horwood. (1993). Insecticide resistance in Myzus persicae (Sulzer) (Hemipetera: Aphididae) in southern Australia, Journal of Entomology society 32, 23–27.
Hopkinson, J., Balzer, J., Fang, C., & Walsh, T. (2023). Insecticide resistance management of Bemisia tabaci (Hemiptera: Aleyrodidae) in Australian cotton–pyriproxyfen, spirotetramat and buprofezin. Pest Management Science, 79(5), 1829–1839.
Hounded, T. A. et al. Insecticide resistance in field populations of Bemisia tabaci (Hemiptera: Aleyrodidae) in West Africa. Pest Manag. Sci. 66, 1181–1185 (2010).
Ishtiaq, M., Saleem, M. A., & Razaq, M. (2012). Monitoring of resistance in Spodoptera exigua (Lepidoptera: Noctuidae) from four districts of the Southern Punjab, Pakistan to four conventional and six new chemistry insecticides. Crop Protection, 33, 13–20.
Ismail, S. M. (2023, March). Laboratory Evaluation of Chlorpyrifos against Resistant and Susceptible Strains of a Moth Pest of Crop Plants, Spodoptera littoralis (Lepidoptera: Noctuidae). In Proceedings of the Zoological Society (Vol. 76, No. 1, pp. 1–5).
Ismail, S.M. 2020. Joint toxic action of spinosad with fenpropathrin and chlorpyrifos and its latent effect on different Egyptian field populations of Spodoptera littoralis. Asian Journal of Biological Sciences 13(4): 2–7.
Khattak, s. d. and Hamed, M., (1993). Effect of different dates of sowing on rapeseed aphid (Brevicoryne brassicae L.) infestation. Proc. Pakistan Con
Mu, W., Wu, K.M., Zhang, W.J., Guo, Y.Y., 2005. Cross-resistance and relative fitness of lambda-cyhalothrin resistant near isogenic lines in Spodoptera exigua (Hubner). Sci. Agric. Sin. 38, 2007e2013.
Munthali DC, Tshegofatso AB (2014). Factors affecting abundance and damage caused by cabbage aphid, Brevicoryne brassicae on four Brassica leafy vegetables: Brassica oleracea var. Acephala, B. ch`inense, B. napus and B. carinata. Open Entomol J 8:1–9.
Mutua, B. K., Dubois, T., Akutse, K. S., Muli, B., Karanja, E. N., & Mutyambai, D. M. (2024). Electrophysiological and behavioural responses of cabbage aphid (Brevicoryne brassicae) to rosemary (Rosmarinus officinalis) volatiles, a potential push plant for vegetable push-pull cropping system.
Naveen, N. C., Chaubey, R., Kumar, D., Rebijith, K. B., Rajagopal, R., Subrahmanyam, B., & Subramanian, S. (2017). Insecticide resistance status in the whitefly, Bemisia tabaci genetic groups Asia-I, Asia-II-1 and Asia-II-7 on the Indian subcontinent. Scientific reports, 7(1), 40634.
Ninsin, K. D. (2004a) Selection for resistance to acetamiprid and various other insecticides in the diamondback moth, Plutella xylostella (L.) (Lep, Plutellidae). J. Appl. Entomol.128, 445–451.
Ninsin, K. D. (2011) Laboratory selection for resistance and susceptibility to acetamiprid in the diamondback moth, Plutella xylostella (L.) from Japan. Ghana Jnl agric. Sci. 44,41–51.
Ninsin, K. D., Mo, J. & Miyata, T. (2000). Decreased susceptibilities of four field populations of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Ypono-meutidae), to acetamiprid. Appl. Entomol. Zool. 35, 591–595.
Patil, S., Sridevi, D., Babu, R. T. and Pushpavathi, B. (2017). Relative efficacy of selected insecticides on cowpea aphid, Aphis craccivora (Koch), Journal of Entomology and Zoological studies, 5, 1603–1607.
Raychaudhuri, D. (1978). Taxonomy and biology of aphids (Homoptera: Aphididae) of Manipur. Ph. D. thesis,
Raychaudhuri, D.N. (1980) (ed.). Aphids of North-East India and Bhutan. Zoological Society, Calcutta, pp. 521.
Sachan, J.N. and Gangwar, S.K. (1990). Seasonal incidence of insect pests of cabbage, cauliflower and knoll-kohl. Indian J. Ent., 52(1): 111–124.
Saeed, S., Ahmad, M., Ahmad, M., Kwon, J.Y., 2007. Insecticidal control of the mealybug Phenacoccus gossypiphyllous (Hemiptera: Pseudococcidae), a new pest of cotton in Pakistan. Entomol. Res. 37, 76e80.
Saleem, M.A., Ahmad, M., Ahmad, M., Aslam, M., 2008. Resistance to selected organochlorine, carbamate and pyrethroid in Spodoptera litura (Lepidoptera: Noctuidae) from Pakistan. J. Econ. Entomol. 101, 1667e1675.
Sethi, A. & Dilawari, V. K. (2008). Spectrum of insecticide resistance in whitefly from upland cotton in Indian subcontinent. J Entomol 5, 138–147 Singh, R., Upadhyay, B.S., Singh, D. and Chaudhary, H.S. (1999).
Shadmany, M., Omar, D. & Muhamad, R. Biotype and insecticide resistance status of Bemisia tabaci populations from Peninsular Malaysia. J. Appl. Entomol. 139, 67–75 (2015).
Sobhy IS, Caulfield JC, Pickett JA, Birkett MA (2020). Sensing the danger signals: cis-Jasmone reduces aphid performance on potato and modulates the magnitude of released volatiles. Front Ecol Evol 7:499.
Sparks, T.C.; Crossthwaite, A.J.; Nauen, R.; Banba, S.; Cordova, D.; Earley, F.; Ebbinghaus-Kintscher, U.; Fujioka, S.; Hirao, A.; Karmon, D.; et al. Insecticides, biologics and nematicides: Updates to IRAC’s mode of action classification—A tool for resistance management. Pestic. Biochem. Physiol. 2020, 167, 104587.
Srivastava, A., Rana, S., Rana, V., & Manuja, S. (2016). Efficacy of quinalphos 25 EC for the management of wheat aphid. Agricultural Science Digest-A Research Journal, 36(4), 337–339.
Tandon, P.L. Bhalla, O.P.and Verma, A.K. (1977). A note on the seasonal population fluctuation of the cabbage aphid on seed cauliflower. Veg. Sci. 4:15–17. University of Calcutta, India, pp. 308 + XVI.
Teran-Vargas, A.P., Garza-Urbina, E., Blanco-Montero, C.A., Perez-Carmona, G., Pellegaud-Rabago, J.M., 1997. Efficacy of new insecticides to control beet armyworm in north eastern Mexico. In: Proc. Beltwide Cotton Conf. National Cotton Council, Memphis, TN, pp. 1030e1031.
Umina, P. A., Edwards, O., Carson, P., VanRooyen, A., & Anderson, A. (2014). High Levels of Resistance to Carbamate and Pyrethroid Chemicals Widespread in Australian Myzus persicae (Hemiptera: Aphididae) Populations. Journal of Economic Entomology, 107(4), 1626–1638
Yu-ping, Z., Yong-yue, L., Ling, Z., & Guang-wen, L. (2010). Life-history traits and population relative fitness of trichlorphon-resistant and-susceptible Bactrocera dorsalis (Diptera: Tephritidae). Psyche: A Journal of Entomology, 2010, 1–8.
Zhang, N.N., C.F. Liu, F. Yang, S.H. Dong, and J.H. Zhao. 2012. Resistance mechanisms to chlorpyrifos and F392W mutation frequencies in the acetylcholine esterase ace1 allele of field populations of the tobacco whitefly, Bemisia tabaci in China. Journal of Insect Science 12(1): 41.

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Evaluation of insecticide resistance in different field populations of Cabbage aphid, Brevicornye brassicae (Hemiptera: Aphididae) to selected organophosphate insecticides

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Insecticides

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Bioassay

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