Current profile of pyrethroid resistance in Rhipicephalus microplus populations sampled from Marathwada region of Maharashtra state, India

Current profile of pyrethroid resistance in Rhipicephalus microplus populations sampled from Marathwada region of Maharashtra state, India | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Current profile of pyrethroid resistance in Rhipicephalus microplus populations sampled from Marathwada region of Maharashtra state, India Sandip Khating, Nitin Jadhav, M. Vijay, Anil Kumar Sharma, Anand Srivastava, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4464602/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 22 Jul, 2024 Read the published version in Parasitology Research → Version 1 posted 7 You are reading this latest preprint version Abstract This study examined the pattern of resistance to widely applied synthetic pyrethroids i.e. cypermethrin and deltamethrin, against larvae of Rhipicephalus microplus ticks sampled from Marathwada region in Maharashtra, India. The study also examined the role of α and β-esterase’s and glutathione-S-transferase (GST) in resistance development. All eight R. microplus isolates tested were resistant to deltamethrin (RL IV), having RR 50 values from 6.88 to 131.26. LPT analysis exhibited the resistance level II deltamethrin resistance in Beed and Hingoli, III in Dharashiv, and IV in Sambhajinagar, Parbhani, Latur, Jalna, and Nanded isolates. The LIT analysis showed that Dharashiv field isolates had the lowest LC 50 value of 229.09 ppm against cypermethrin, while Sambhajinagar field isolates had the highest at 489.78 ppm. The RR 50 ranged from 1145.45 to 2448.9. Seven isolates were level I resistant to cypermethrin while the Jalna isolate was level II resistant. In larvae treated with deltamethrin and cypermethrin, the activity of α and β-esterase enzymes increased significantly compared to control groups. The enzyme ratios in treated larvae ranged from 0.7533 to 1.7023 for α-esterase and 0.7434 to 3.2054 for β-esterase. The Hingoli isolate treated with cypermethrin exhibited the highest α-esterase activity (903.261), whereas, Sambhajinagar isolate had the highest GST enzyme ratio (2.8224) after deltamethrin exposure. When exposed to cypermethrin, the Hingoli isolate showed the highest GST enzyme ratio, 2.0832. Rhipicephalus microplus Resistance Deltamethrin Cypermethrin Metabolic enzymes Maharashtra Figures Figure 1 Introduction Ticks and tick-borne diseases (TTBDs) cause the fourth most livestock infections impacting about eighty percent cattle population of the world. Internationally, TTBD in cattle cost US $ 13.9 to 18.7 billion (de Castro 1997 ) and US $ 498.7 million in India (Minjauw and McLeod 2003 ). Many tick species infest the livestock and exert adverse effect on animal health and their products in the world. Rhipicephalus microplus is a highly significant tick species in India that transmits Anaplasma marginale and Babesia bigemina , serious pathogens of animal diseases (Ghosh et al., 2007 ). Each fully fed female of R. microplus reduces Bos taurus bovine body weight by 1.37 ± 0.25g and daily milk output by 8.9 ± 2.1 ml (Jonsson et al. 1998 ; Rodrigues and Leite 2013 ). As per the estimate of Furlong et al. (1996), an infestation of 105 ticks per cow can reduce milk production by 23% per day and 40 ticks per animal can reduce weight by 20 kg per year and cost of hides by 20–30%. Most of the livestock owners in India adopt application of chemical acaricides for rapid and affordable suppression of tick infestations. However, the extensive utilization of acaricides with inaccurate dilution, inappropriate application techniques, persistent and indiscriminate usage, and excessive dosage has come up with the resistance emergence in field ticks (Ghosh et al. 2006 ; Abbas et al. 2014 ). Chemical acaricides also pollute the environment and leave toxic residues in milk and meat (Graf et al. 2004 ). De León et al. ( 2020 ) found that R. microplus is the sixth most resistant arthropod worldwide due to the widespread usage of acaricides. Acaricidal pressure on animals as a tick management component in different regions of India has caused low to high resistance level against pyrethroids and other commonly available classes of acaricides (Sharma et al. 2012 ; Shyma et al. 2012 ; Kumar et al. 2021 ). Various studies on acaricide resistance mechanisms like target site changes, behavioural changes, metabolic enzyme or ATP binding cassette transporter detoxification, and a decrease in chemical acaricide penetration have been done (Hemingway et al. 2004 ; Rosario-Cruz et al. 2009 ; Fular et al. 2020 ). Esterase enzyme pathways have been found in pyrethroids-resistant R. microplus (Baffi et al. 2008 ). Similarly, arthropods have been found to have an enhanced expression of GST gene transcription or higher enzyme activity, indicating the involvement of GST in the metabolism of both endogenous and exogenous substances (Hernandez et al. 2018 ). According to FAO ( 2004 ), regular screening of commonly used acaricides is critical for avoiding tick resistance, and decreasing acaricide resistance spread. However, no comprehensive research has been conducted to determine the acaricidal resistance status and to contribution of the role of metabolic enzymes in the ticks of Marathwada region of Maharashtra. In the current study, this region has been explored for identification of resistant tick population for effective execution of chemical tick control programme in Marathwada region. Materials and methods Study area Maharashtra stands third-largest state of India encompassing 307,713 km 2 and 9.36% of total area of the country. The state covers 15°35'-22°02'N and 72°36'-80°54'E on the globe. Maharashtra's tropical monsoonal climate produces 1,181 mm of rainfall annually, with 75% of it falling during the southwest monsoon season from June to September. The state is geographically partitioned into five distinct regional divisions: Western Maharashtra, Marathwada, Konkan, Vidarbha and North Maharashtra (Fig. 1 ). Out of these regions, the Marathwada region mostly relies on rainfall for agriculture and has limited access to irrigation facilities. Consequently, the abundant availability of resources in the Marathwada region has resulted in substantial growth in the dairy industry, serving as the primary catalyst for income and job creation for farmers. The cattle breeds that are most favoured in the Marathwada region include Red Kandhari, Deoni, Holstein Friesian, Jersey and Gir. As for buffalo breeds, the popular ones include Murrah, Jaffrabadi, Marathwadi, and Pandharpuri. All breeds, with significant differences, are experiencing tick infestations (Shirsikar et al. 2023 ). Collection and processing of field isolates The Marathwada region of Maharashtra state is notable for hosting several tick species and tick-borne diseases mostly due to suitable climatic conditions that facilitate tick growth and survival round the year. Furthermore, the rising practice of maintaining crossbred animals to enhance milk production and by-product yields has led to an increase in the occurrence of tick infestation and the diseases they transmit. Despite the numerous limitations of chemical acaricides, farmers in the region continue to employ them for tick control. The study was conducted in response to concerns from farmers in the region regarding the ineffective treatment and or reduced protection of chemical acaricides to control the infestation of Rhipicephalus microplus ticks despite multiple administrations. A total of eight districts in the Marathwada region of Maharashtra state, namely Parbhani, Hingoli, Nanded, Jalana, Chhatrapati Sambhajinagar, Beed, Latur, and Dharashiv, were chosen to collect ticks from both unorganized and organized sectors and were coded as PAR, HIN, NAN, JAL, SAM, BEE, LAT and DHR, respectively. The ticks were collected by a two-stage stratified sampling procedure, chosen for its adaptability and capacity to maintain a consistent equilibrium between statistical precision and cost-effectiveness. As a result, a consistent categorization method was employed to divide each area into categories, ensuring a precise and representative sampling of ticks in the district. A survey was conducted to gather information on the frequency, mode of application, and duration of protection provided by commonly used acaricides against reinfestation. Careful examinations of the animal shed were carried out during the collection procedure in order to identify and pinpoint hiding places, such as crevices, loose bricks, and other ground debris. The engorged adult female ticks of R. microplus were obtained either by manually removing them from naturally-infested calves or by directly collecting them from the animals on the same day they dropped. The ticks were rinsed in normal water and then dried on tissue paper. The obtained ticks from eight areas were identified at the Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, (MAFSU), Parbhani. The collected ticks were kept in desiccators which were placed in a BOD incubator at 28 ◦ C and relative humidity of 85 ± 5% for oviposition. After 12–14 days, the tick eggs were gathered together and placed in individual vials. The vials were then covered with muslin cloth and secured with rubber bands. The eggs were let to hatch after ten days. The larval based test i.e. larval immersion (LIT) and larval packet tests (LPT) utilized larvae that were around 10 to 14 days old. To conduct biochemical research, the larvae were preserved at -80 o C (Fular et al. 2018 ). Preparation of working concentrations of acaricides The commercial formulations are designed to include certain proprietary components and synergists that may enhance the effectiveness of acaricides against ticks. Whereas, technical grade acaricides consist of refined chemicals and are more sensitive for detection of reduction in the susceptibility of field ticks (Ghosh et al. 2017 ). For the assay, deltamethrin and cypermethrin compounds (Technical grade) were fetched from Sigma Aldrich, USA, were utilized to characterise the resistance. The solvents such as acetone and methanol were used for preparation of stock solution of deltamethrin (1000ppm) and cypermethrin (5000ppm), respectively. From the stock solution, a series of working concentrations in the multiples of LC 95 value of acaricides (x, 2x, 3x, 4x, 5x, 6x, 7x etc.) were developed by mixing in distilled water where ‘x’is the LC 95 of reference susceptible strain of R. microplus . Using LC 95 x2, the discriminating concentration (DC) of an acaricide was calculated. Reference tick strains A reference strain susceptible to deltamethrin (LC 50 = 0.02 ppm and LC 95 = 21.29 ppm) and cypermethrin (LC 50 = 0.20 and LC 95 = 26.5 ppm were used (Shyma et al. 2019 ). For LPT, deltamethrin: LC 50 = 11.8 ppm, LC 95 = 35.5 ppm; cypermethrin: LC 50 = 242.5 ppm, LC 95= 350.7 ppm was used Sharma et al. ( 2012 ). Based on RF values, R. microplus field isolates were categorized into susceptible = RF < 1.4, level I = RF 1.5–5.0, level II = RF 5.1–25.0, level III = RF 26–40 and level IV = RF 41or above. In-vitro bioassays for larvicidal efficacy of the acaricides Larval Immersion test (LIT) LIT was executed as described by Shaw ( 1966 ). In short, 100–120 larvae were immersed in 200µl of varying acaricide concentrations in a 1.5ml centrifuge tube. The larvae were immersed for 10 minutes to observe larvicidal activity. After 10 minutes, a pipette withdrew all the fluid, and the tubes were dried by gently rotating the filter paper strips. The tubes were tied with muslin cloth with the help of rubber band. Control groups were immersed in the solvents in the same way. The concentration of individual solvent was maintained below 5% in each working solution. For each acaricide concentration, three replicates were employed. Subsequently, the larvae were placed in desiccators and a BOD incubator at 85 ± 5% RH and 28◦C. By counting dead and alive larvae after 24 hours of incubation, each acaricide concentration was assessed for larvicidal efficacy. Larval packet test (LPT) The LPT was designed according to the Food and Agriculture Organization procedure (FAO 1971 ). The 0.5 ml of different acaricides was used to impregnate a larval packet prepared from 7.0 cm x7.0 cm filter paper (Whatman No. 541). Impregnated filter sheets at 37°C were incubated for 30 minutes to evaporate solvents. After complete evaporation of the solvents, filter paper sheet was diagonally folded in half and was closed with adhesive tape at one side. The tick larvae (100–150) were placed through open-end, sealed with tape and packets containing larvae were kept at 28◦C and 85 ± 5% RH in a BOD incubator. Similarly, with the suitable solvents, control packets were also treated. Larvicidal activity was tested three times per acaricide concentration. After one day, the packets were unsealed in light and the pedal movement of the larvae was used to count the dead. Quantification of enzymatic activity of esterase’s and glutathione-S-transferases (GST) Initial exposure to chemical acaricide LC 50 concentrations was done on larvae. Each isolate, treated and untreated (control group), was sampled at 10 mg in 1.5 mL eppendorf tubes. Then, the larval tube was plunged in liquid nitrogen and mashed with a motorized pestle. Next, 1 ml of chilled 20 mM Phosphate buffered saline (PBS) with a pH of 7.2 was introduced and subjected to sonication for a duration of 5 minutes, following a cycle of 5 seconds on and 10 seconds off at an amplitude of 30%. The larval homogenates underwent centrifugation at a speed of 12,000 rpm at 4 0 C for duration of 15 minutes. The supernatant was utilized to investigate biochemical alterations. Some of the homogenates were preserved at a temperature of -20°C for later use. The protein contents (g/mL) in the homogenates were obtained by calculating various quantities of bovine serum albumin, as described by Bradford in 1976. The quantification of esterase’s and glutathione-S-transferases (GST) was performed using a tick-specific standardized procedure established by Fular et al. ( 2020 ). To quantify esterase’s, 20µL of the homogenate was combined with 200µL of a 30 mM stock solution of α- or β-naphthyl acetate in phosphate buffer (0.02 M, pH 7.2). This mixture was then placed in triplicate in a 96-well microtiter plate. Instead of using larval homogenate, distilled water was employed in the control. The reaction mixture was left to react for 15 minutes at ambient temperature. Following incubation, the fast blue solution (50µL) with 15mg fast blue dissolved in distilled water (1.5 mL) and 5% SDS (3.5 mL) in sodium phosphate buffer (0.1M at pH 7), was introduced into wells. The plates were thereafter placed at room temperature for 5 minutes to facilitate the occurrence of a change in colour, and the absorbance was quantified at a wavelength of 570 nm using an ELISA reader. The enzyme level was quantified as the amount of product produced per minute per milligram of protein, measured in micromoles. For GST quantification, 10µL of larval homogenate and 200µL of a working solution were added in duplicate. The working solution contained 2.5 mL reduced glutathione (10 mM GSH) in phosphate buffer (pH 6.5) and 125µL of 63 mM chloroditrobenzene. All of the reagents were recently prepared. The plates were placed in a controlled environment for 20 minutes without light, and the amount of light absorbed at a wavelength of 340 nm was recorded with an ELISA reader. The estimation of GST concentration was done by Lambert-Beer's law. A = εCL. Where, ε = Extinction coefficient of product (3-(2-chloro-4-nitrophenyl)-glutathione) = 4.39 mM − 1 , L = Length of path (in this case was 6 mm), C represents variables of concentration, A represents absorbance at 340 nm after 20 minutes. Statistical analysis After testing different concentrations of acaricides, the entomological parameters have been analyzed applying probit analysis (Finney, 1962) applying GraphPad Prism 8 statistical computer application (GraphPad Software, San Diego, California, USA) to estimate LC 50 and LC 95 doses. The resistance ratio of field isolates of R. microplus was determined by dividing their LC 50 value by a reference susceptible strain. The data obtained from the biochemical assay was analysed using analysis of variance (ANOVA) to identify any statistically significant variations among the groups. Results Following the LIT protocol, different isolates were characterized using the deltamethrin and it was found that the RR 50 (RL) values ranged from 6745 to 34591.5, indicating resistance to deltamethrin in all field isolates. The JAL isolate had the highest LC 50 of 691.83 ppm, while the HIN isolate demonstrated a relatively lower i.e., 66.07 ppm. Out of the eight samples studied using LPT revealed the resistance level II in BEE and HIN, level III resistance in DHR isolates and resistance level IV in five isolates from the SAM, PAR, LAT, JAL, and NAN (Table 1 ). This study found considerable deltamethrin resistance in both bioassays. It could be due to widespread deltamethrin use to control mosquitoes and other pests in agriculture and veterinary has led to a greater threshold of resistance to deltamethrin in cattle ticks in this region. Table 1 Resistance pattern to deltamethrin in field population of R. microplus using the LIT and LPT Tick isolates LPT LPT Slope LC 50 (ppm) LC 95 (ppm) RR 50 (RL) Slope LC 50 (ppm) LC 95 (ppm) RR 50 (RL) BEE 2.395 134.90 645.65 6745(IV) 3.37 275.42 831.76 23.34(II) DHA 3.2922 151.36 478.63 7568(IV) 4.6748 316.23 707.95 26.80(III) SAM 4.7214 257.04 630.96 12852(IV) 8.0299 512.86 831.76 43.46(IV) HIN 3.0748 66.07 223.87 3303.5(IV) 2.7931 81.18 316.23 6.88(II) PAR 3.682 204.17 575.44 10208.5(IV) 8.3195 645.65 1000.00 54.72(IV) LAT 3.0826 199.53 691.83 9976.5(IV) 10.308 1023.29 1479.11 86.72(IV) JAL 10.058 691.83 1023.29 34591.5(IV) 13.288 1548.82 2041.74 131.26(IV) NAN 3.9156 275.42 724.44 13771(IV) 10.381 741.31 1071.52 62.82(IV) RR 50 (median) resistance ratio, RL resistance level (susceptible [S] = RR < 1.4; level I: 1.5 < RR < 5; level II: 5.1 < RR < 25; level III: 26 < RR 41 In addition, cypermethrin is next to deltamethrin which is widely used for tick management in different parts of the country. Present findings using cypermethrin against larvae of R. microplus revealed level IV resistance to cypermethrin in all the isolates in which SAM isolate showed the highest RR 50 (2448.9), while the DHR isolate had lowest (1145.45) (Table 2 ). Table 2 Resistance pattern to cypermethrin in field population of R. microplus using the LIT and LPT Tick Isolates LPT LIT Slope LC 50 (ppm) LC 95 (ppm) RR 50 (RL) Slope LC 50 (ppm) LC 95 (ppm) RR 50 (RL) BEE 4.5178 446.68 1023.29 2233.4 (IV) 2.7871 549.54 2089.30 2.27 (I) DHA 2.6542 229.09 954.99 1145.45(IV) 2.6948 478.63 1949.84 1.97 (I) SAM 2.7889 489.78 1862.09 2448.9 (IV) 3.6655 891.25 2511.89 3.68 (I) HIN 3.4548 446.68 1318.26 2233.4 (IV) 2.6958 501.19 2041.74 2.07 (I) PAR 2.3855 426.58 2089.30 2132.9 (IV) 3.1997 794.33 2630.27 3.28 (I) LAT 2.6011 309.03 1318.26 1545.15 (IV) 2.5129 575.44 2570.40 2.37 (I) JAL 2.777 478.63 1862.09 2393.15 (IV) 12.209 3630.78 4897.79 14.97 (II) NAN 3.6375 478.63 1348.96 2393.15 (IV) 2.6633 489.78 2041.74 2.02 (I) RR50 (median) resistance ratio, RL resistance level (susceptible [S] = RR < 1.4; level I: 1.5 < RR < 5; level II: 5.1 < RR < 25; level III: 26 < RR 41 The present study aimed to investigate the involvement of metabolic enzymes responsible for resistance development to SP acaricides. The results of the study demonstrated that R. microplus larvae, when exposed to deltamethrin and cypermethrin at LC 50 concentrations, exhibited elevated levels of esterase enzymes. Deltamethrin exposed larvae had α and β-esterase activity ranging from 252.931 to 425.255 and 136.268 to 271.715, respectively. The NAN isolate showed the highest value for α (1.7023) and β-esterase (3.2054) enzyme ratios, with a resistance factor (RF) of 62.82. In the HIN isolate (RF = 2.07), α- and β-naphthyl acetate had the greatest enzyme ratios, 3.7610 and 3.8343, respectively (Table 3 , 4 ). Table 3 α-esterase and β-esterase activity in various field population of R. microplus exposed to deltamethrin Tick Isolates Group RR 50 α-esterase activity (nanomolar/min.µg of protein) Enzyme ratio β-esterase activity (nanomolar/min.µg of protein) Enzyme ratio BEE Treated 23.34 361.996 a 0.9163 271.715 0.9473 Control 395.049 b 286.826 OSM Treated 26.80 342.654 0.8882 254.388 0.9330 Control 385.802 272.651 AUR Treated 43.46 339.300 a 1.0326 245.664 a 1.0418 Control 328.600 b 235.799 b HIN Treated 6.88 284.809 a 1.0098 162.943 a 1.0647 Control 282.058 b 153.043 b PAR Treated 54.72 252.931 a 1.1085 136.268 a 1.0796 Control 228.179 b 126.215 b LAT Treated 86.72 341.458 a 0.7533 250.935 a 0.7434 Control 453.254 b 337.554 b JAL Treated 131.26 413.761 0.9771 212.795 a 1.5759 Control 423.478 135.034 b NAN Treated 62.82 425.255 a 1.7023 254.453 a 3.2054 Control 249.808 b 79.383 b a, b Superscript shows significant difference between treated and control group for respective field isolates Table 4 α-esterase and β-esterase activity in various field population of R. microplus exposed to cypermethrin Tick Isolates Group RR 50 α-esterase activity (nanomolar/min.µg of protein) Enzyme ratio β-esterase activity (nanomolar/min.µg of protein) Enzyme ratio BEE Treated 2.27 328.047 a 0.9658 288.986 a 0.9952 Control 339.674 b 290.389 b OSM Treated 1.97 444.504 a 1.3305 216.845 a 0.7869 Control 334.093 b 275.581 b AUR Treated 3.68 297.025 0.9466 201.733 a 0.9711 Control 313.773 207.731 b HIN Treated 2.07 903.261 a 3.7610 557.868 a 3.8343 Control 240.167 b 145.494 b PAR Treated 3.28 256.491 1.0558 145.494 a 0.9859 Control 242.941 147.582 b LAT Treated 2.37 221.034 a 0.7816 73.499 a 0.7125 Control 282.784 b 103.158 b JAL Treated 14.97 294.176 a 1.3763 161.275 a 1.5261 Control 213.736 b 105.675 b NAN Treated 2.02 316.720 a 2.0082 193.234 a 1.4151 Control 157.713 b 136.552 b a,b Superscripts shows significant difference between treated and control group for respective districts Similarly, the role of GST activity in eight R. microplus field isolates was quantified and the deltamethrin exposed larvae had a GST activity range of 80.250-714.030, while the control group had a range of 64.547-615.728 nanomolar/min µg. The HIN isolate showed the highest GST activity with an RF of 2.07, followed by SAM (3.68), and DHR (1.97). After treatment with LC 50 of cypermethrin, the GST activity ranged from 0.3711 to 2.0832. The HIN isolate indicated the highest GST enzyme ratio (2.0832), followed by SAM (1.2433) and DHR (1.2132) while the lowest GST enzyme ratio (0.3711) occurred in JAL isolates (Table 5 ). Table 5 GST activity in field isolates of R. microplus exposed to deltamethrin and cypermethrin Tick Isolates Group Deltamethrin treated larvae Cypermethrin treated larvae Resistance factor GST activity (nanomolar/min.µg of protein) Enzyme ratio Resistance factor GST activity (nanomolar/min.µg of protein) Enzyme ratio BED Treated 23.34 335.034 1.0144 2.27 128.594 0.6153 Control 330.278 208.989 OSM Treated 26.80 324.330 0.9067 1.97 121.956 1.2132 Control 357.700 100.521 AUR Treated 43.46 216.226 a 2.8224 3.68 80.250 1.2433 Control 76.609 b 64.547 HIN Treated 6.88 248.046a 1.5295 2.07 714.030 a 2.0832 Control 162.174 b 342.752 b PAR Treated 54.72 175.067 a 0.9645 3.28 229.970 0.5868 Control 181.520 b 391.885 LAT Treated 86.72 359.119 a 0.9811 2.37 81.152 a 0.6007 Control 366.024 b 135.096 b JAL Treated 131.26 269.601 a 1.9630 14.97 228.473 0.3711 Control 137.341 b 615.728 NAN Treated 62.82 352.266 a 2.2904 2.02 369.180 0.7289 Control 153.802 b 506.515 a,b superscripts shows significant difference between treated and control group for respective field isolates Discussion Due to favourable tick growth and survival circumstances throughout the year, tick species and tick-borne diseases are common in Maharashtra. The increased practice of rearing crossbred animals for milk production has increased the rate of tick infestation. R. microplus is the most prevalent species in Maharashtra (Maske et al. 1997 ). Farmers in the region continue employ chemical acaricides to manage ticks, despite their drawbacks. The shorter lifetime and higher rates of reproduction of R. microplus , which need frequent application of synthetic chemicals, have played a major role in the worldwide emergence of resistance (Guerrero et al. 2014 ). Additionally, an easy availability of acaricides to farmers, insufficient application and utilization of acaricides with different brand names but the same active ingredient, and the lack of a monitoring system have also contributed to resistance in India (Kumar et al. 2020 ). Among the available chemical acaricides, synthetic pyrethroids are the most frequently promoted acaricides which make over 25% of the global pesticide market and have been widely used to manage cattle ticks for 15 years (Kumar et al. 2020 ). The accessible availability, high safety margin, and indiscriminate use of these compounds created widespread resistance and the biggest resistance factors (> 100) compared to other chemical acaricides worldwide (Van Wyk et al. 2016 ). In this study, resistance in field isolates was estimated using LC 50 instead of LC 95 , as Sindhu et al. ( 2022 ) mentioned that the LC 50 is statistically more effective than the LC 99 in any bioassay, estimating the RF 99 using the LC 99 may introduce inaccuracy, hence it is recommended to consider the RF 50 . Earlier, Robertson et al. ( 2007 ) found that the use of LC 99 in LPT bioassay resulted in an inaccurate assessment of the resistance factor. All eight field isolates showed level IV resistance to deltamethrin. Sharma et al. ( 2012 ) reported a maximum (95.7) and highest RF (288.72) in the Sikar isolate (Rajasthan) for deltamethrin, while Ahanger et al. ( 2015 ) reported the highest RF in R. (B.) microplus obtained from Jammu. Consistent with the findings of the current investigation, Sharma et al. ( 2012 ) documented the most significant degree of resistance to deltamethrin in R. (B.) microplus across six different agro-climatic zones in India. Furthermore, reports showed various pictures of deltamethrin resistance in R. (B.) microplus from various regions of India have been published (Chigure et al. 2018 ; Godara et al. 2019 ; Shakya et al. 2021 ; Fular et al., 2021; Gupta et al.2023). All field isolates were exposed to identical agroclimatic conditions, however resistance levels varied. Gaur et al. ( 2016 ) also found different levels of deltamethrin resistance in field isolates collected from the same agro-climatic zones identifying Agroha isolates as susceptible and Tohana isolate as resistant. The factors such as prolonged exposure to deltamethrin, improper use of acaricides, and mutation may contribute to variations in resistance levels. A significant disparity in the resistance level of deltamethrin across tick populations in field isolates with same environmental conditions was also reported (Sindhu et al. 2022 ). Alonso-Díaz, et al. (2006) suggested that the various factors, such as tick migration between farms, animal breed, favourable conditions for the survival of ticks in their non-parasitic phase, the presence or absence of refuges, and the intensity of acaricide use attributed to the variation in resistance levels. SP resistance is often referred to as family resistance, as ticks develop resistance to many chemicals within the same group simultaneously (Miller1988). In accordance with observations of present study, Kumar et al. ( 2017 ) found cypermethrin resistance in Chittoor (RL = II), Prakasam and YSR Kadapa (RL = I) of Andhra Pradesh. In this investigation, LPT showed increased deltamethrin resistance and sluggish cypermethrin resistance against all field isolates. Earlier investigations by Sharma et al. ( 2012 ), Ahanger and Ghosh (2015), Godara (2019) found comparatively lower level cypermethrin than deltamethrin. Several factors including target site modifications, individual behaviour toward acaricides changes, detoxification by metabolic enzymes or ATP binding cassette transporters and less penetration can reduce arthropods' chemical acaricide susceptibility (Hemingway et al. 2004 ; Rosario-Cruz et al. 2009 ). Arthropods like R. microplus build resistance by detoxification process which is mostly carried out by metabolic enzymes. Earlier reports indicated the potential involvement of general esterase’s in the metabolic resistance of the Indian field tick population to synthetic pyrethroids (Shyma et al. 2012 ; Rinesh Kumar et al. 2013). The metabolism and detoxification of foreign compounds and native substances depend on glutathione-S-transferases (GSTs). Similarly, Enayati et al. (2005) also stressed the importance of GST in SP acaricide metabolism. Previous studies have documented the involvement of GST in the breakdown of both endogenous and exogenous substances through either an elevated expression of the GST gene or an increase in enzyme activity (Hernandez et al. 2018 ). The current findings corroborate the previously reported data on increased level of esterase of resistant R. microplus ticks procured from Haryana, as shown by Gaur et al. ( 2016 ). In a similar vein, the research carried out by Ghosh et al. ( 2015 ) demonstrated a noteworthy escalation in the levels of esterase, monooxygenase, and GST activity in the resistant R. microplus isolates obtained from Bihar in Eastern India. In another study, Kumar et al. ( 2021 ) noted the significantly higher esterase activity in the tick populations sampled from different states of India compared to the reference IVRI-I strain ranging from 0.096 ± 0.045 to 7.893 ± 0.302. Conclusion The resistance development status of ticks procured from eight districts of Marathwada region of Maharashtra clearly indicates that the SP compounds (deltamethrin and cypermethrin) are not working efficiently for tick control. Ticks infested animals in Marathwada region responded differently to these acaricides, and so region-specific control strategies in such a large state need to be worked out. The use of SP compounds should be regulated in this region to prevent further establishment of resistant populations and other newer drugs for tick management should be introduced. Besides, more studies on acaricide resistance in other divisions of the state needs to be carried out to work out present status of tick resistance. Effective management of the current situation and the preservation of a secure environment require collaborative efforts including all stakeholders. Declarations Author contribution Conceptualisation: GC, NJ. Data curation: AS. Formal analysis: SK. Investigation: SK, MV, SKu, MFMFS. Methodology: GC, AD. Supervision: GC, AS. Writing—original draft: SK, MR. Writing — review and editing: AKS, MR, GC Data availability Not applicable. Ethics approval Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Acknowledgements The authors are grateful to Associate Dean, College of Veterinary & Animal Sciences, MAFSU, Parbhani for providing facilities to carry out research work. References Abbas RZ, Zaman MA, Colwell DD, Gilleard J, Iqbal Z (2014) Acaricide resistance in cattle ticks and approaches to its management: the state of play. Vet Parasitol 203(1–2): 6–20. https://doi.org/10.1016/j.vetpar.2014.03.006 Ahanger RR, Godara R, Katoch R, Yadav A, Bhutyal ADS, Katoch M, Bader MA (2015) Deltamethrin resistance in field populations of Rhipicephalus ( Boophilus ) microplus (Acari: Ixodidae) in Jammu and Kashmir, India.Exp Appl Acarol 67:467–475. Alonso-Diaz MA, Rodriguez-Vivas RI, Fragoso-Sanchez H, Rosario-Cruz R (2006) Ixodicide resistance of the the Boophilus microplus tick to ixodicides. Archivos de medicina veterinaria 38(2):105–113.DOI: 10.4067/S0301-732X2006000200003 Baffi MA, De Souza GRL, De Sousa CS, Ceron CR, Bonetti AM (2008) Esterase enzymes involved in pyrethroid and organophosphate resistance in a Brazilian population of Riphicephallus ( Boophilus ) microplus (Acari, Ixodidae). Mol Biochem Parasitol 160(1):70–73. https://doi.org/10.1016/j.molbiopara.2008.03.009 Bradford M M (1976) A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principles of protein dye binding. Anal Biochem 72: 248–254. Chigure GM, Sharma AK, Kumar S, Fular A, Sagar SV, Nagar G, Ghosh S (2018) Role of metabolic enzymes in conferring resistance to synthetic pyrethroids, organophosphates, and phenylpyrazole compounds in Rhipicephalus microplus . Int J Acarol 44(1):28–34. https://doi.org/10.1080/01647954.2017.1400588 de Castro JJ (1997) Sustainable tick and tickborne disease control in livestock improvement in developing countries. Vet Parasitol 71(2–3):77–97. https://doi.org/10.1016/S0304-4017(97)00033-2 De León AAP, Mitchell RD III, Watson DW (2020) Ectoparasites of cattle. Vet Clin North Am Food Anim Pract 36(1):173–185. https://doi.org/10.1016/j.cvfa.2019.12.004 FAO (1971) Recommended methods for the detection and measurement of resistance of agricultural pests to pesticides – tentative method for larvae of cattle tick, Boophilus microplus spp . FAO method No. 7. FAO Plant Protect. Bull. 19: 15–18. FAO (2004) Resistance Management and Integrated Parasite Control in Ruminants: Guidelines. Module 1. Ticks: Acaricide Resistance: Diagnosis, Management and Prevention. Animal Production and Health Division, Rome. FAO: pp. 25–77. Finny DJ (1962) Probit Analysis- A Statistical Treatment of the Response Curve. Cambridge University Press, Cambridgepp. 1–318. Fular A, Gupta S, Sharma AK, Kumar S, Upadhaya D, Shakya M, Ghosh S (2020) Standardization of tick specific biochemical tools for estimation of esterases, monooxygenases and glutathione S-transferase for characterization of acaricide resistance. Pestic Biochem Phy 164:130–139.DOI: 10.1016/j.pestbp.2020.01.008 Fular A, Sharma AK, Kumar S, Nagar G, Chigure G, Ray DD, Ghosh S (2018) Establishment of a multi-acaricide resistant reference tick strain (IVRI-V) of Rhipicephalus microplus . Ticks Tick Borne Dis9(5):1184–1191. https://doi.org/10.1016/j.ttbdis.2018.04.014 Gaur RS, Sangwan AK, Sangwan N, Kumar S (2016) Acaricide resistance in Rhipicephalus ( Boophilus ) microplus and Hyalommaanatolicum collected from Haryana and Rajasthan states of India. Exp Appl Acarol69:487–500. DOI: 10.1007/s10493-016-0046-1 Ghosh S, Azhahianambi P, De la Fuente J (2006) Control of ticks of ruminants, with special emphasis on livestock farming systems in India: present and future possibilities for integrated control—a review. Exp Appl Acarol 40:49–66. DOI: 10.1007/s10493-006-9022-5 Ghosh S, Bansal GC, Gupta SC, Ray D, Khan MQ, Irshad H, Ahmed JS (2007) Status of tick distribution in Bangladesh, India and Pakistan. Parasitol Res 101:207–216. DOI: 10.1007/s00436-007-0684-7 Ghosh S, Gupta S, Kumar KA, Sharma AK, Kumar S, Nagar G, Ray D (2017) Characterization and establishment of a reference deltamethrin and cypermethrin resistant tick line (IVRI-IV) of Rhipicephalus ( Boophilus ) microplus . Pestic Biochem Phys 138:66–70. https://doi.org/10.1016/j.pestbp.2017.03.002 Ghosh S, Kumar R, Nagar G, Kumar S, Sharma AK, Srivastava A, Saravanan BC (2015) Survey of acaricides resistance status of Rhipiciphalus ( Boophilus ) microplus collected from selected places of Bihar, an eastern state of India. Ticks Tick Borne Dis 6(5):668–675. https://doi.org/10.1016/j.ttbdis.2015.05.013 Ghosh SS, Azhahianambi PA, Yadav MP (2007) Upcoming and future strategies of tick control: a review. J Vect Borne Dis 44(2):79–89 Godara R, Katoch R, Rafiqi SI, Yadav A, Nazim K, Sharma R, Katoch M (2019) Synthetic pyrethroid resistance in Rhipicephalus (Boophilus) microplus ticks from north-western Himalayas, India. Trop Anim Health Prod 51:1203–1208. DOI: 10.1007/s11250-019-01810-8 Graf JF, Gogolewski R, Leach-Bing N, Sabatini GA, Molento MB, Bordin EL, Arantes GJ (2004) Tick control: an industry point of view. Parasitol 129(S1):S427-S442. DOI: https://doi.org/10.1017/S0031182004006079 Guerrero F, Pérez de León A, Rodriguez-Vivas R, Jonsson N, Miller R, Andreotti R. E. N. A. T. O, Roe RM (2014) Acaricide research and development, resistance and resistance monitoring. Sonenshine DE, Roe RM, editors, 2:353–374. Gupta S, Sangwan N, Sangwan AK, Mann S, Gupta S, Kumar A, Kumar S (2023) Understanding the resistance mechanisms of Rhipicephalus microplus ticks to synthetic pyrethroids and organophosphates in south-west regions of Haryana, North India. Pestic Biochem Physiol 196: 105634. https://doi.org/10.1016/j.pestbp.2023.105634 Hemingway J, Hawkes NJ, McCarroll L, Ranson H (2004) The molecular basis of insecticide resistance in mosquitoes. Insect Biochem Mol Biol 34(7):653–665. https://doi.org/10.1016/j.ibmb.2004.03.018 Hernandez EP, Kusakisako K, Talactac MR, Galay RL, Hatta T, Fujisaki K, Tanaka T (2018) Glutathione S-transferases play a role in the detoxification of flumethrin and chlorpyrifos in Haemaphysalis longicornis .Parasit Vectors 11:1–14.DOI: 10.1186/s13071-018-3044-9 Jonsson NN, Mayer DG, Matschoss AL, Green PE, Ansell J (1998) Production effects of cattle tick ( Boophilus microplus ) infestation of high yielding dairy cows. Vet Parasitol 78(1):65–77. https://doi.org/10.1016/S0304-4017(98)00118-6 Rinesh Kumar, Nagar G, Sharma AK, Kumar S, Ray DD, Chaudhuri P, Ghosh S (2013) Survey of pyrethroids resistance in Indian isolates of Rhipicephalus ( Boophilus ) microplus : identification of C190A mutation in the domain II of the para-sodium channel gene. Acta Trop 125(2): 237–245. https://doi.org/10.1016/j.actatropica.2012.10.006 Kumar R, Sharma AK, Ghosh S (2020) Menace of acaricide resistance in cattle tick, Rhipicephalus microplus in India: Status and possible mitigation strategies. Vet Parasitol 278, 108993. https://doi.org/10.1016/j.vetpar.2019.108993 Kumar S, Sharma AK, Kumar B, Shakya M, Patel JA, Kumar B, Ghosh S (2021) Characterization of deltamethrin, cypermethrin, coumaphos and ivermectin resistance in populations of Rhipicephalus microplus in India and efficacy of an antitick natural formulation prepared from Ageratum conyzoides . Ticks Tick Borne Dis 12(6):101818. https://doi.org/10.1016/j.ttbdis.2021.101818 Kumar SS, Rayulu VC, Rao KS, Kumar NV (2017) Acaricidal resistance in Rhipicephalus ( Boophilus ) Microplus ticks infesting cattle of Andhra Pradesh. J Entomol Zool Stud 5(6):580–584. Maske DK, Sardey MR, Bhilegaonkar NG (1997). Prevalence of ixodid ticks on cattle in Vidarbha region of Maharashtra. Indian I Anim Sci67(9): 768–769. Miller TA (1988) Mechanisms of resistance to pyrethroid insecticides. Parasitology Today 4(7):S8-S12. DOI:https://doi.org/10.1016/0169-4758(88)90080-4 Minjauw B, McLeod A (2003) Tick-borne diseases and poverty: the impact of ticks and tick-borne diseases on the livelihoods of small-scale and marginal livestock owners in India and eastern and southern Africa. Tick-borne diseases and poverty: the impact of ticks and tick-borne diseases on the livelihoods of small-scale and marginal livestock owners in India and eastern and southern Africa.124-pp Singh O, Bhardwaj P, Singh J (2017) Distribution of Lightning Casualties over Maharashtra, India. J Ind Geophys Union 21(5): 415–424 Robertson JL,Russell RM, Priesler HK,Savin NE (2007) Bioassay with arthropods, second edn. CRC Press, Boca Raton. https://doi.org/10.1201/9781420004045 Rodrigues DS, Leite RC (2013) Economic impact of Rhipicephalus ( Boophilus ) microplus : estimate of decreased milk production on a dairy farm. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 65:1570–1572. Rosario-Cruz R, Almazan C, Miller R, Dominguez-Garcia DI, Hernandez-Ortiz R (2009) Front Biosci (Landmark Ed) 14(7): 2657–65. doi: 10.2741/3403 . Shakya M, Singh M, Kumar S, Jayraw AK, Jatav GP, Agrawal V, Jamara N (2021) Efficacy of commercially available chemical compound used against management of tick infestation in Mhow, Madhya Pradesh. Int J Trop Insect Sci 41:2469–2475. DOI: 10.1007/s42690-020-00424-0 Sharma AK, Kumar R, Kumar S, Nagar G, Singh NK, Rawat SS, Ghosh S (2012) Deltamethrin and cypermethrin resistance status of Rhipicephalus ( Boophilus ) microplus collected from six agro-climatic regions of India. Vet Parasitol 188(3–4):337–345. https://doi.org/10.1016/j.vetpar.2012.03.050 Shaw RD (1966) Culture of an organophosphorus-resistant strain of Boophilus microplus (Can.) and an assessment of its resistance spectrum. Bull Ento Res 56(3):389–405. DOI: https://doi.org/10.1017/S0007485300056480 Shyma KP, Kumar S, Sharma AK, Ray DD, Ghosh S (2012) Acaricide resistance status in Indian isolates of Hyalomma anatolicum . Exp Appl Acarol 58:471–481. DOI: 10.1007/s10493-012-9592-3 Shyma KP, Singh V, Gupta JP (2019) Susceptibility Level of Ornithodoros spp (Acari: Argasidae) to Some Commercial Synthetic Pyrethroids in North Gujarat. Int J Livest Res 9(3):62–67. DOI: 10.5455/ijlr.20180522085604 Shirsikar PM, Narladkar BW, Jadhav ND, Rajurkar SR, Jadhav SG, Channa GR, Gaikwad SS, Chigure GM (2023) Morphological characterization and prevalence of hard ticks infesting cattle of Marathwada Region, Maharashtra state, India. J Exp Zool India26(2). 10.51470/jez.2023.26.2.2575 Sindhu ZUD, Naseer MU, Raza A, Aslam B, Ahmad J, Abbas RZ, Khattak B (2022) Resistance to Cypermethrin Is Widespread in Cattle Ticks ( Rhipicephalus microplus ) in the Province of Punjab, Pakistan: In Vitro Diagnosis of Acaricide Resistance. Pathogens 11(11):1293. https://doi.org/10.3390/pathogens11111293 Van Wyk RD, Baron S, Maritz-Olivier C (2016) An integrative approach to understanding pyrethroid resistance in Rhipicephalus microplus and R . decoloratus ticks. Ticks Tick Borne Dis 7(4):586–594. DOI: 10.1016/j.ttbdis.2016.01.007 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 22 Jul, 2024 Read the published version in Parasitology Research → Version 1 posted Reviewers agreed at journal 05 Jun, 2024 Reviewers agreed at journal 04 Jun, 2024 Reviewers agreed at journal 04 Jun, 2024 Reviewers invited by journal 04 Jun, 2024 Editor assigned by journal 29 May, 2024 Submission checks completed at journal 27 May, 2024 First submitted to journal 23 May, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4464602","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":310708394,"identity":"12655af5-1177-44e7-9230-5b0338f87690","order_by":0,"name":"Sandip Khating","email":"","orcid":"","institution":"MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sandip","middleName":"","lastName":"Khating","suffix":""},{"id":310708397,"identity":"3fc5cfbc-1e39-4049-a74e-51d161de8381","order_by":1,"name":"Nitin Jadhav","email":"","orcid":"","institution":"MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Nitin","middleName":"","lastName":"Jadhav","suffix":""},{"id":310708398,"identity":"0b0d53fc-1f42-4d79-832e-9e17bfbc15f5","order_by":2,"name":"M. Vijay","email":"","orcid":"","institution":"DBT-National Institute of Animal Biotechnology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"M.","middleName":"","lastName":"Vijay","suffix":""},{"id":310708399,"identity":"e654e253-263a-4304-aeb4-946ffda659b8","order_by":3,"name":"Anil Kumar Sharma","email":"","orcid":"","institution":"Uttaranchal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Anil","middleName":"Kumar","lastName":"Sharma","suffix":""},{"id":310708400,"identity":"6318e98e-0219-4d0c-a61a-58e7a46f7a21","order_by":4,"name":"Anand Srivastava","email":"","orcid":"","institution":"DBT-National Institute of Animal Biotechnology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Anand","middleName":"","lastName":"Srivastava","suffix":""},{"id":310708401,"identity":"a42d35e8-030f-4b6f-b7a8-9a199ca24522","order_by":5,"name":"Suresh Jadhao","email":"","orcid":"","institution":"Nagpur Veterinary College, MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Suresh","middleName":"","lastName":"Jadhao","suffix":""},{"id":310708402,"identity":"167b5d89-cd5a-4c8d-9a1c-64bef0635764","order_by":6,"name":"Sachin Kumar","email":"","orcid":"","institution":"Constituent Government College (MJP Rohilkhand University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sachin","middleName":"","lastName":"Kumar","suffix":""},{"id":310708403,"identity":"bcbdcf14-bb56-4348-81b1-56c021c9c653","order_by":7,"name":"Shrikant Kalwaghe","email":"","orcid":"","institution":"MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shrikant","middleName":"","lastName":"Kalwaghe","suffix":""},{"id":310708404,"identity":"42caf2a4-cf01-4372-87c6-62aeaaa972ca","order_by":8,"name":"M. F.M.F. Siddiqui","email":"","orcid":"","institution":"MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"M.","middleName":"F.M.F.","lastName":"Siddiqui","suffix":""},{"id":310708405,"identity":"da0a5efd-ab79-4b4e-9d1a-a9bcbf78fde4","order_by":9,"name":"Mahima Narawade","email":"","orcid":"","institution":"MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mahima","middleName":"","lastName":"Narawade","suffix":""},{"id":310708406,"identity":"7336d5cf-53d4-46aa-ad38-152b8ae35eee","order_by":10,"name":"Ankush Dhabale","email":"","orcid":"","institution":"MAFSU","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ankush","middleName":"","lastName":"Dhabale","suffix":""},{"id":310708407,"identity":"9ebf990d-31e8-4086-805e-29e1fe3b7510","order_by":11,"name":"Gajanan Chigure","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBElEQVRIiWNgGAWjYHCCBDBpwAymbBgYmA+AuMRrSWNgYEsgqAUCoGoOQ7XgAfLuBx5+Lqi5I2/Ozn5NuqLifDR/G/szCYaCOzi1GJ5JSJaeceyZ4c5mnjLJM2du5844xmMmwWDwDLeWhoQEaR62wwkGh3nSJBvbbuc23O9hA2o5jFtL/4Pk3zz/YFr+ncudfwzkMDxa5CUS0qR520Ba2I9JNjYcyN1wjMEMrxYDiQdp1rx9hw03HOZhtmw4lpy78RiPsUUCPlv6c5Jv83w7LG9w/vjDmw01drnzjrE/vPHhDx5bDvAkQJk8SNGXgEUp3JYG9gNQJvsDPOpGwSgYBaNgJAMArLZZUhZZAlAAAAAASUVORK5CYII=","orcid":"","institution":"MAFSU","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Gajanan","middleName":"","lastName":"Chigure","suffix":""}],"badges":[],"createdAt":"2024-05-23 06:21:32","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4464602/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4464602/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00436-024-08306-x","type":"published","date":"2024-07-22T16:15:02+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":58025628,"identity":"cce28621-574a-4780-9692-07977a682ca2","added_by":"auto","created_at":"2024-06-10 06:29:44","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":294664,"visible":true,"origin":"","legend":"\u003cp\u003eMap showing locations of tick collection from Mara thwada region (Aurangabad division) of Maharashtra state of India (Source: Singh et al. 2017)\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-4464602/v1/e90dacf8c51320c99e5a24ac.png"},{"id":61596939,"identity":"ca42418d-141e-4851-82e0-0590d0203825","added_by":"auto","created_at":"2024-08-01 17:30:44","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1294020,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4464602/v1/c14e0aa8-f774-4993-a625-33cf58e9d959.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Current profile of pyrethroid resistance in Rhipicephalus microplus populations sampled from Marathwada region of Maharashtra state, India","fulltext":[{"header":"Introduction","content":"\u003cp\u003eTicks and tick-borne diseases (TTBDs) cause the fourth most livestock infections impacting about eighty percent cattle population of the world. Internationally, TTBD in cattle cost US\u003cspan\u003e$\u003c/span\u003e 13.9 to 18.7\u0026nbsp;billion (de Castro \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1997\u003c/span\u003e) and US\u003cspan\u003e$\u003c/span\u003e 498.7\u0026nbsp;million in India (Minjauw and McLeod \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Many tick species infest the livestock and exert adverse effect on animal health and their products in the world. \u003cem\u003eRhipicephalus microplus\u003c/em\u003e is a highly significant tick species in India that transmits \u003cem\u003eAnaplasma marginale\u003c/em\u003e and \u003cem\u003eBabesia bigemina\u003c/em\u003e, serious pathogens of animal diseases (Ghosh et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Each fully fed female of \u003cem\u003eR. microplus\u003c/em\u003e reduces \u003cem\u003eBos taurus\u003c/em\u003e bovine body weight by 1.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25g and daily milk output by 8.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1 ml (Jonsson et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Rodrigues and Leite \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). As per the estimate of Furlong et al. (1996), an infestation of 105 ticks per cow can reduce milk production by 23% per day and 40 ticks per animal can reduce weight by 20 kg per year and cost of hides by 20\u0026ndash;30%.\u003c/p\u003e \u003cp\u003eMost of the livestock owners in India adopt application of chemical acaricides for rapid and affordable suppression of tick infestations. However, the extensive utilization of acaricides with inaccurate dilution, inappropriate application techniques, persistent and indiscriminate usage, and excessive dosage has come up with the resistance emergence in field ticks (Ghosh et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Abbas et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Chemical acaricides also pollute the environment and leave toxic residues in milk and meat (Graf et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). De Le\u0026oacute;n et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) found that \u003cem\u003eR. microplus\u003c/em\u003e is the sixth most resistant arthropod worldwide due to the widespread usage of acaricides. Acaricidal pressure on animals as a tick management component in different regions of India has caused low to high resistance level against pyrethroids and other commonly available classes of acaricides (Sharma et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Shyma et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Kumar et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eVarious studies on acaricide resistance mechanisms like target site changes, behavioural changes, metabolic enzyme or ATP binding cassette transporter detoxification, and a decrease in chemical acaricide penetration have been done (Hemingway et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Rosario-Cruz et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Fular et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Esterase enzyme pathways have been found in pyrethroids-resistant \u003cem\u003eR. microplus\u003c/em\u003e (Baffi et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Similarly, arthropods have been found to have an enhanced expression of GST gene transcription or higher enzyme activity, indicating the involvement of GST in the metabolism of both endogenous and exogenous substances (Hernandez et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). According to FAO (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2004\u003c/span\u003e), regular screening of commonly used acaricides is critical for avoiding tick resistance, and decreasing acaricide resistance spread. However, no comprehensive research has been conducted to determine the acaricidal resistance status and to contribution of the role of metabolic enzymes in the ticks of Marathwada region of Maharashtra. In the current study, this region has been explored for identification of resistant tick population for effective execution of chemical tick control programme in Marathwada region.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy area\u003c/h2\u003e \u003cp\u003eMaharashtra stands third-largest state of India encompassing 307,713 km\u003csup\u003e2\u003c/sup\u003e and 9.36% of total area of the country. The state covers 15\u0026deg;35'-22\u0026deg;02'N and 72\u0026deg;36'-80\u0026deg;54'E on the globe. Maharashtra's tropical monsoonal climate produces 1,181 mm of rainfall annually, with 75% of it falling during the southwest monsoon season from June to September. The state is geographically partitioned into five distinct regional divisions: Western Maharashtra, Marathwada, Konkan, Vidarbha and North Maharashtra (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Out of these regions, the Marathwada region mostly relies on rainfall for agriculture and has limited access to irrigation facilities. Consequently, the abundant availability of resources in the Marathwada region has resulted in substantial growth in the dairy industry, serving as the primary catalyst for income and job creation for farmers. The cattle breeds that are most favoured in the Marathwada region include Red Kandhari, Deoni, Holstein Friesian, Jersey and Gir. As for buffalo breeds, the popular ones include Murrah, Jaffrabadi, Marathwadi, and Pandharpuri. All breeds, with significant differences, are experiencing tick infestations (Shirsikar et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eCollection and processing of field isolates\u003c/h2\u003e \u003cp\u003eThe Marathwada region of Maharashtra state is notable for hosting several tick species and tick-borne diseases mostly due to suitable climatic conditions that facilitate tick growth and survival round the year. Furthermore, the rising practice of maintaining crossbred animals to enhance milk production and by-product yields has led to an increase in the occurrence of tick infestation and the diseases they transmit. Despite the numerous limitations of chemical acaricides, farmers in the region continue to employ them for tick control. The study was conducted in response to concerns from farmers in the region regarding the ineffective treatment and or reduced protection of chemical acaricides to control the infestation of \u003cem\u003eRhipicephalus microplus\u003c/em\u003e ticks despite multiple administrations. A total of eight districts in the Marathwada region of Maharashtra state, namely Parbhani, Hingoli, Nanded, Jalana, Chhatrapati Sambhajinagar, Beed, Latur, and Dharashiv, were chosen to collect ticks from both unorganized and organized sectors and were coded as PAR, HIN, NAN, JAL, SAM, BEE, LAT and DHR, respectively. The ticks were collected by a two-stage stratified sampling procedure, chosen for its adaptability and capacity to maintain a consistent equilibrium between statistical precision and cost-effectiveness. As a result, a consistent categorization method was employed to divide each area into categories, ensuring a precise and representative sampling of ticks in the district.\u003c/p\u003e \u003cp\u003eA survey was conducted to gather information on the frequency, mode of application, and duration of protection provided by commonly used acaricides against reinfestation. Careful examinations of the animal shed were carried out during the collection procedure in order to identify and pinpoint hiding places, such as crevices, loose bricks, and other ground debris. The engorged adult female ticks of \u003cem\u003eR. microplus\u003c/em\u003e were obtained either by manually removing them from naturally-infested calves or by directly collecting them from the animals on the same day they dropped. The ticks were rinsed in normal water and then dried on tissue paper. The obtained ticks from eight areas were identified at the Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, (MAFSU), Parbhani.\u003c/p\u003e \u003cp\u003eThe collected ticks were kept in desiccators which were placed in a BOD incubator at 28\u003csup\u003e◦\u003c/sup\u003eC and relative humidity of 85\u0026thinsp;\u0026plusmn;\u0026thinsp;5% for oviposition. After 12\u0026ndash;14 days, the tick eggs were gathered together and placed in individual vials. The vials were then covered with muslin cloth and secured with rubber bands. The eggs were let to hatch after ten days. The larval based test i.e. larval immersion (LIT) and larval packet tests (LPT) utilized larvae that were around 10 to 14 days old. To conduct biochemical research, the larvae were preserved at -80\u003csup\u003eo\u003c/sup\u003eC (Fular et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of working concentrations of acaricides\u003c/h2\u003e \u003cp\u003eThe commercial formulations are designed to include certain proprietary components and synergists that may enhance the effectiveness of acaricides against ticks. Whereas, technical grade acaricides consist of refined chemicals and are more sensitive for detection of reduction in the susceptibility of field ticks (Ghosh et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). For the assay, deltamethrin and cypermethrin compounds (Technical grade) were fetched from Sigma Aldrich, USA, were utilized to characterise the resistance. The solvents such as acetone and methanol were used for preparation of stock solution of deltamethrin (1000ppm) and cypermethrin (5000ppm), respectively. From the stock solution, a series of working concentrations in the multiples of LC\u003csub\u003e95\u003c/sub\u003e value of acaricides (x, 2x, 3x, 4x, 5x, 6x, 7x etc.) were developed by mixing in distilled water where \u0026lsquo;x\u0026rsquo;is the LC\u003csub\u003e95\u003c/sub\u003eof reference susceptible strain of \u003cem\u003eR. microplus\u003c/em\u003e. Using LC\u003csub\u003e95\u003c/sub\u003ex2, the discriminating concentration (DC) of an acaricide was calculated.\u003c/p\u003e \u003cp\u003e \u003cb\u003eReference tick strains\u003c/b\u003e \u003c/p\u003e \u003cp\u003eA reference strain susceptible to deltamethrin (LC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.02 ppm and LC\u003csub\u003e95\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;21.29 ppm) and cypermethrin (LC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.20 and LC\u003csub\u003e95\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;26.5 ppm were used (Shyma et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). For LPT, deltamethrin: LC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;11.8 ppm, LC\u003csub\u003e95\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;35.5 ppm; cypermethrin: LC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;242.5 ppm, LC\u003csub\u003e95=\u003c/sub\u003e350.7 ppm was used Sharma et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Based on RF values, \u003cem\u003eR. microplus\u003c/em\u003e field isolates were categorized into susceptible\u0026thinsp;=\u0026thinsp;RF\u0026thinsp;\u0026lt;\u0026thinsp;1.4, level I\u0026thinsp;=\u0026thinsp;RF 1.5\u0026ndash;5.0, level II\u0026thinsp;=\u0026thinsp;RF 5.1\u0026ndash;25.0, level III\u0026thinsp;=\u0026thinsp;RF 26\u0026ndash;40 and level IV\u0026thinsp;=\u0026thinsp;RF 41or above.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIn-vitro\u003c/b\u003e \u003cb\u003ebioassays for larvicidal efficacy of the acaricides\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eLarval Immersion test (LIT)\u003c/h2\u003e \u003cp\u003eLIT was executed as described by Shaw (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1966\u003c/span\u003e). In short, 100\u0026ndash;120 larvae were immersed in 200\u0026micro;l of varying acaricide concentrations in a 1.5ml centrifuge tube. The larvae were immersed for 10 minutes to observe larvicidal activity. After 10 minutes, a pipette withdrew all the fluid, and the tubes were dried by gently rotating the filter paper strips. The tubes were tied with muslin cloth with the help of rubber band. Control groups were immersed in the solvents in the same way. The concentration of individual solvent was maintained below 5% in each working solution. For each acaricide concentration, three replicates were employed. Subsequently, the larvae were placed in desiccators and a BOD incubator at 85\u0026thinsp;\u0026plusmn;\u0026thinsp;5% RH and 28◦C. By counting dead and alive larvae after 24 hours of incubation, each acaricide concentration was assessed for larvicidal efficacy.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eLarval packet test (LPT)\u003c/h2\u003e \u003cp\u003eThe LPT was designed according to the Food and Agriculture Organization procedure (FAO \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1971\u003c/span\u003e). The 0.5 ml of different acaricides was used to impregnate a larval packet prepared from 7.0 cm x7.0 cm filter paper (Whatman No. 541). Impregnated filter sheets at 37\u0026deg;C were incubated for 30 minutes to evaporate solvents. After complete evaporation of the solvents, filter paper sheet was diagonally folded in half and was closed with adhesive tape at one side. The tick larvae (100\u0026ndash;150) were placed through open-end, sealed with tape and packets containing larvae were kept at 28◦C and 85\u0026thinsp;\u0026plusmn;\u0026thinsp;5% RH in a BOD incubator. Similarly, with the suitable solvents, control packets were also treated. Larvicidal activity was tested three times per acaricide concentration. After one day, the packets were unsealed in light and the pedal movement of the larvae was used to count the dead.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eQuantification of enzymatic activity of esterase\u0026rsquo;s and glutathione-S-transferases (GST)\u003c/h2\u003e \u003cp\u003eInitial exposure to chemical acaricide LC\u003csub\u003e50\u003c/sub\u003e concentrations was done on larvae. Each isolate, treated and untreated (control group), was sampled at 10 mg in 1.5 mL eppendorf tubes. Then, the larval tube was plunged in liquid nitrogen and mashed with a motorized pestle. Next, 1 ml of chilled 20 mM Phosphate buffered saline (PBS) with a pH of 7.2 was introduced and subjected to sonication for a duration of 5 minutes, following a cycle of 5 seconds on and 10 seconds off at an amplitude of 30%. The larval homogenates underwent centrifugation at a speed of 12,000 rpm at 4\u003csup\u003e0\u003c/sup\u003eC for duration of 15 minutes. The supernatant was utilized to investigate biochemical alterations. Some of the homogenates were preserved at a temperature of -20\u0026deg;C for later use. The protein contents (g/mL) in the homogenates were obtained by calculating various quantities of bovine serum albumin, as described by Bradford in 1976.\u003c/p\u003e \u003cp\u003eThe quantification of esterase\u0026rsquo;s and glutathione-S-transferases (GST) was performed using a tick-specific standardized procedure established by Fular et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). To quantify esterase\u0026rsquo;s, 20\u0026micro;L of the homogenate was combined with 200\u0026micro;L of a 30 mM stock solution of α- or β-naphthyl acetate in phosphate buffer (0.02 M, pH 7.2). This mixture was then placed in triplicate in a 96-well microtiter plate. Instead of using larval homogenate, distilled water was employed in the control. The reaction mixture was left to react for 15 minutes at ambient temperature. Following incubation, the fast blue solution (50\u0026micro;L) with 15mg fast blue dissolved in distilled water (1.5 mL) and 5% SDS (3.5 mL) in sodium phosphate buffer (0.1M at pH 7), was introduced into wells. The plates were thereafter placed at room temperature for 5 minutes to facilitate the occurrence of a change in colour, and the absorbance was quantified at a wavelength of 570 nm using an ELISA reader. The enzyme level was quantified as the amount of product produced per minute per milligram of protein, measured in micromoles.\u003c/p\u003e \u003cp\u003eFor GST quantification, 10\u0026micro;L of larval homogenate and 200\u0026micro;L of a working solution were added in duplicate. The working solution contained 2.5 mL reduced glutathione (10 mM GSH) in phosphate buffer (pH 6.5) and 125\u0026micro;L of 63 mM chloroditrobenzene. All of the reagents were recently prepared. The plates were placed in a controlled environment for 20 minutes without light, and the amount of light absorbed at a wavelength of 340 nm was recorded with an ELISA reader. The estimation of GST concentration was done by Lambert-Beer's law. A\u0026thinsp;=\u0026thinsp;εCL. Where, ε\u0026thinsp;=\u0026thinsp;Extinction coefficient of product (3-(2-chloro-4-nitrophenyl)-glutathione)\u0026thinsp;=\u0026thinsp;4.39 mM\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, L\u0026thinsp;=\u0026thinsp;Length of path (in this case was 6 mm), C represents variables of concentration, A represents absorbance at 340 nm after 20 minutes.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAfter testing different concentrations of acaricides, the entomological parameters have been analyzed applying probit analysis (Finney, 1962) applying GraphPad Prism 8 statistical computer application (GraphPad Software, San Diego, California, USA) to estimate LC\u003csub\u003e50\u003c/sub\u003e and LC\u003csub\u003e95\u003c/sub\u003e doses. The resistance ratio of field isolates of \u003cem\u003eR. microplus\u003c/em\u003e was determined by dividing their LC\u003csub\u003e50\u003c/sub\u003e value by a reference susceptible strain. The data obtained from the biochemical assay was analysed using analysis of variance (ANOVA) to identify any statistically significant variations among the groups.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eFollowing the LIT protocol, different isolates were characterized using the deltamethrin and it was found that the RR\u003csub\u003e50\u003c/sub\u003e (RL) values ranged from 6745 to 34591.5, indicating resistance to deltamethrin in all field isolates. The JAL isolate had the highest LC\u003csub\u003e50\u003c/sub\u003e of 691.83 ppm, while the HIN isolate demonstrated a relatively lower i.e., 66.07 ppm. Out of the eight samples studied using LPT revealed the resistance level II in BEE and HIN, level III resistance in DHR isolates and resistance level IV in five isolates from the SAM, PAR, LAT, JAL, and NAN (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). This study found considerable deltamethrin resistance in both bioassays. It could be due to widespread deltamethrin use to control mosquitoes and other pests in agriculture and veterinary has led to a greater threshold of resistance to deltamethrin in cattle ticks in this region.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eResistance pattern to deltamethrin in field population of \u003cem\u003eR. microplus\u003c/em\u003e using the LIT and LPT\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTick isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eLPT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e \u003cp\u003eLPT\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSlope\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLC\u003csub\u003e50\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLC\u003csub\u003e95\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e (RL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSlope\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eLC\u003csub\u003e50\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLC\u003csub\u003e95\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e (RL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBEE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.395\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e134.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e645.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6745(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e275.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e831.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e23.34(II)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDHA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.2922\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e151.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e478.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7568(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4.6748\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e316.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e707.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e26.80(III)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSAM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.7214\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e257.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e630.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12852(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e8.0299\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e512.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e831.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e43.46(IV)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHIN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.0748\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e66.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e223.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3303.5(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.7931\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e81.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e316.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e6.88(II)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePAR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.682\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e204.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e575.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10208.5(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e8.3195\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e645.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1000.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e54.72(IV)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.0826\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e199.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e691.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9976.5(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.308\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1023.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1479.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e86.72(IV)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eJAL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.058\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e691.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1023.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34591.5(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13.288\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1548.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2041.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e131.26(IV)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNAN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.9156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e275.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e724.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13771(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.381\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e741.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1071.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e62.82(IV)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e (median) resistance ratio, RL resistance level (susceptible [S]\u0026thinsp;=\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;1.4; level I: 1.5\u0026thinsp;\u0026lt;\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;5; level II: 5.1\u0026thinsp;\u0026lt;\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;25; level III: 26\u0026thinsp;\u0026lt;\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;40; level IV: RR\u0026thinsp;\u0026gt;\u0026thinsp;41\u003c/p\u003e \u003cp\u003eIn addition, cypermethrin is next to deltamethrin which is widely used for tick management in different parts of the country. Present findings using cypermethrin against larvae of \u003cem\u003eR. microplus\u003c/em\u003e revealed level IV resistance to cypermethrin in all the isolates in which SAM isolate showed the highest RR\u003csub\u003e50\u003c/sub\u003e (2448.9), while the DHR isolate had lowest (1145.45) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eResistance pattern to cypermethrin in field population of \u003cem\u003eR. microplus\u003c/em\u003e using the LIT and LPT\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTick Isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eLPT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e \u003cp\u003eLIT\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSlope\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLC\u003csub\u003e50\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLC\u003csub\u003e95\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e(RL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSlope\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eLC\u003csub\u003e50\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLC\u003csub\u003e95\u003c/sub\u003e (ppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e(RL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBEE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.5178\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e446.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1023.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2233.4 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.7871\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e549.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2089.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.27 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDHA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.6542\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e229.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e954.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1145.45(IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.6948\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e478.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1949.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.97 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSAM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.7889\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e489.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1862.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2448.9 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.6655\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e891.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2511.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.68 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHIN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.4548\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e446.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1318.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2233.4 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.6958\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e501.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2041.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.07 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePAR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.3855\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e426.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2089.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2132.9 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.1997\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e794.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2630.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.28 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.6011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e309.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1318.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1545.15 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.5129\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e575.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2570.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.37 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eJAL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.777\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e478.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1862.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2393.15 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e12.209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3630.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4897.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e14.97 (II)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNAN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.6375\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e478.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1348.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2393.15 (IV)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.6633\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e489.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2041.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.02 (I)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003eRR50 (median) resistance ratio, RL resistance level (susceptible [S]\u0026thinsp;=\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;1.4; level I: 1.5\u0026thinsp;\u0026lt;\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;5; level II: 5.1\u0026thinsp;\u0026lt;\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;25; level III: 26\u0026thinsp;\u0026lt;\u0026thinsp;RR\u0026thinsp;\u0026lt;\u0026thinsp;40; level IV: RR\u0026thinsp;\u0026gt;\u0026thinsp;41\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe present study aimed to investigate the involvement of metabolic enzymes responsible for resistance development to SP acaricides. The results of the study demonstrated that \u003cem\u003eR. microplus\u003c/em\u003e larvae, when exposed to deltamethrin and cypermethrin at LC\u003csub\u003e50\u003c/sub\u003e concentrations, exhibited elevated levels of esterase enzymes. Deltamethrin exposed larvae had α and β-esterase activity ranging from 252.931 to 425.255 and 136.268 to 271.715, respectively. The NAN isolate showed the highest value for α (1.7023) and β-esterase (3.2054) enzyme ratios, with a resistance factor (RF) of 62.82. In the HIN isolate (RF\u0026thinsp;=\u0026thinsp;2.07), α- and β-naphthyl acetate had the greatest enzyme ratios, 3.7610 and 3.8343, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eα-esterase and β-esterase activity in various field population of \u003cem\u003eR. microplus\u003c/em\u003e exposed to deltamethrin\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTick Isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eα-esterase activity (nanomolar/min.\u0026micro;g of protein)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eEnzyme ratio\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eβ-esterase activity (nanomolar/min.\u0026micro;g of protein)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eEnzyme ratio\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eBEE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e23.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e361.996\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9163\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e271.715\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9473\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e395.049\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e286.826\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eOSM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e26.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e342.654\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.8882\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e254.388\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9330\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e385.802\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e272.651\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAUR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e43.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e339.300\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0326\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e245.664\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0418\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e328.600\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e235.799\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eHIN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e6.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e284.809\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0098\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e162.943\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0647\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e282.058\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e153.043\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePAR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e54.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e252.931\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.1085\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e136.268\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0796\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e228.179\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e126.215\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e86.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e341.458\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.7533\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e250.935\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.7434\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e453.254\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e337.554\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eJAL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e131.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e413.761\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9771\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e212.795\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.5759\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e423.478\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e135.034\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNAN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e62.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e425.255\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.7023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e254.453\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.2054\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e249.808\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e79.383\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e\u003csup\u003ea, b\u003c/sup\u003e Superscript shows significant difference between treated and control group for respective field isolates\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eα-esterase and β-esterase activity in various field population of \u003cem\u003eR. microplus\u003c/em\u003e exposed to cypermethrin\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTick Isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRR\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eα-esterase activity (nanomolar/min.\u0026micro;g of protein)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eEnzyme ratio\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eβ-esterase activity (nanomolar/min.\u0026micro;g of protein)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eEnzyme ratio\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eBEE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e328.047\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9658\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e288.986\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9952\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e339.674\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e290.389\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eOSM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e444.504\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.3305\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e216.845\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.7869\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e334.093\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e275.581\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAUR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e297.025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9466\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e201.733\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9711\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e313.773\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e207.731\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eHIN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e903.261\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.7610\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e557.868\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.8343\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e240.167\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e145.494\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePAR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e256.491\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0558\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e145.494\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9859\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e242.941\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e147.582\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e221.034\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.7816\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e73.499\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.7125\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e282.784\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e103.158\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eJAL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e14.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e294.176\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.3763\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e161.275\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.5261\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e213.736\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e105.675\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNAN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e316.720\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.0082\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e193.234\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.4151\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e157.713\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e136.552\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e\u003csup\u003ea,b\u003c/sup\u003eSuperscripts shows significant difference between treated and control group for respective districts\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eSimilarly, the role of GST activity in eight \u003cem\u003eR. microplus\u003c/em\u003e field isolates was quantified and the deltamethrin exposed larvae had a GST activity range of 80.250-714.030, while the control group had a range of 64.547-615.728 nanomolar/min \u0026micro;g. The HIN isolate showed the highest GST activity with an RF of 2.07, followed by SAM (3.68), and DHR (1.97). After treatment with LC\u003csub\u003e50\u003c/sub\u003e of cypermethrin, the GST activity ranged from 0.3711 to 2.0832. The HIN isolate indicated the highest GST enzyme ratio (2.0832), followed by SAM (1.2433) and DHR (1.2132) while the lowest GST enzyme ratio (0.3711) occurred in JAL isolates (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eGST activity in field isolates of \u003cem\u003eR. microplus\u003c/em\u003e exposed to deltamethrin and cypermethrin\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTick Isolates\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003eDeltamethrin treated larvae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c8\" namest=\"c6\"\u003e \u003cp\u003eCypermethrin treated larvae\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eResistance factor\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGST activity (nanomolar/min.\u0026micro;g of protein)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eEnzyme ratio\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eResistance factor\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGST activity (nanomolar/min.\u0026micro;g of protein)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eEnzyme ratio\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eBED\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e23.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e335.034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.0144\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e128.594\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.6153\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e330.278\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e208.989\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eOSM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e26.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e324.330\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9067\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e121.956\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.2132\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e357.700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100.521\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eAUR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e43.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e216.226\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.8224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e80.250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.2433\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e76.609\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e64.547\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eHIN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e6.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e248.046a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.5295\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e714.030\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.0832\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e162.174\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e342.752\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePAR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e54.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e175.067\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9645\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e3.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e229.970\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.5868\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e181.520\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e391.885\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e86.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e359.119\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.9811\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e81.152\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.6007\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e366.024\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e135.096\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eJAL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e131.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e269.601\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.9630\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e14.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e228.473\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.3711\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e137.341\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e615.728\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNAN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e62.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e352.266\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.2904\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e2.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e369.180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.7289\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e153.802\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e506.515\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003e\u003csup\u003ea,b\u003c/sup\u003esuperscripts shows significant difference between treated and control group for respective field isolates\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eDue to favourable tick growth and survival circumstances throughout the year, tick species and tick-borne diseases are common in Maharashtra. The increased practice of rearing crossbred animals for milk production has increased the rate of tick infestation. \u003cem\u003eR. microplus\u003c/em\u003e is the most prevalent species in Maharashtra (Maske et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). Farmers in the region continue employ chemical acaricides to manage ticks, despite their drawbacks. The shorter lifetime and higher rates of reproduction of \u003cem\u003eR. microplus\u003c/em\u003e, which need frequent application of synthetic chemicals, have played a major role in the worldwide emergence of resistance (Guerrero et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Additionally, an easy availability of acaricides to farmers, insufficient application and utilization of acaricides with different brand names but the same active ingredient, and the lack of a monitoring system have also contributed to resistance in India (Kumar et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong the available chemical acaricides, synthetic pyrethroids are the most frequently promoted acaricides which make over 25% of the global pesticide market and have been widely used to manage cattle ticks for 15 years (Kumar et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The accessible availability, high safety margin, and indiscriminate use of these compounds created widespread resistance and the biggest resistance factors (\u0026gt;\u0026thinsp;100) compared to other chemical acaricides worldwide (Van Wyk et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In this study, resistance in field isolates was estimated using LC\u003csub\u003e50\u003c/sub\u003e instead of LC\u003csub\u003e95\u003c/sub\u003e, as Sindhu et al. (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) mentioned that the LC\u003csub\u003e50\u003c/sub\u003e is statistically more effective than the LC\u003csub\u003e99\u003c/sub\u003e in any bioassay, estimating the RF\u003csub\u003e99\u003c/sub\u003e using the LC\u003csub\u003e99\u003c/sub\u003e may introduce inaccuracy, hence it is recommended to consider the RF\u003csub\u003e50\u003c/sub\u003e. Earlier, Robertson et al. (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) found that the use of LC\u003csub\u003e99\u003c/sub\u003e in LPT bioassay resulted in an inaccurate assessment of the resistance factor.\u003c/p\u003e \u003cp\u003eAll eight field isolates showed level IV resistance to deltamethrin. Sharma et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) reported a maximum (95.7) and highest RF (288.72) in the Sikar isolate (Rajasthan) for deltamethrin, while Ahanger et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) reported the highest RF in \u003cem\u003eR. (B.) microplus\u003c/em\u003e obtained from Jammu. Consistent with the findings of the current investigation, Sharma et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) documented the most significant degree of resistance to deltamethrin in \u003cem\u003eR. (B.) microplus\u003c/em\u003e across six different agro-climatic zones in India. Furthermore, reports showed various pictures of deltamethrin resistance in \u003cem\u003eR. (B.) microplus\u003c/em\u003e from various regions of India have been published (Chigure et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Godara et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Shakya et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Fular et al., 2021; Gupta et al.2023).\u003c/p\u003e \u003cp\u003eAll field isolates were exposed to identical agroclimatic conditions, however resistance levels varied. Gaur et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) also found different levels of deltamethrin resistance in field isolates collected from the same agro-climatic zones identifying Agroha isolates as susceptible and Tohana isolate as resistant. The factors such as prolonged exposure to deltamethrin, improper use of acaricides, and mutation may contribute to variations in resistance levels. A significant disparity in the resistance level of deltamethrin across tick populations in field isolates with same environmental conditions was also reported (Sindhu et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Alonso-D\u0026iacute;az, et al. (2006) suggested that the various factors, such as tick migration between farms, animal breed, favourable conditions for the survival of ticks in their non-parasitic phase, the presence or absence of refuges, and the intensity of acaricide use attributed to the variation in resistance levels.\u003c/p\u003e \u003cp\u003eSP resistance is often referred to as family resistance, as ticks develop resistance to many chemicals within the same group simultaneously (Miller1988). In accordance with observations of present study, Kumar et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) found cypermethrin resistance in Chittoor (RL\u0026thinsp;=\u0026thinsp;II), Prakasam and YSR Kadapa (RL\u0026thinsp;=\u0026thinsp;I) of Andhra Pradesh. In this investigation, LPT showed increased deltamethrin resistance and sluggish cypermethrin resistance against all field isolates. Earlier investigations by Sharma et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), Ahanger and Ghosh (2015), Godara (2019) found comparatively lower level cypermethrin than deltamethrin.\u003c/p\u003e \u003cp\u003eSeveral factors including target site modifications, individual behaviour toward acaricides changes, detoxification by metabolic enzymes or ATP binding cassette transporters and less penetration can reduce arthropods' chemical acaricide susceptibility (Hemingway et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Rosario-Cruz et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Arthropods like \u003cem\u003eR. microplus\u003c/em\u003e build resistance by detoxification process which is mostly carried out by metabolic enzymes. Earlier reports indicated the potential involvement of general esterase\u0026rsquo;s in the metabolic resistance of the Indian field tick population to synthetic pyrethroids (Shyma et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Rinesh Kumar et al. 2013). The metabolism and detoxification of foreign compounds and native substances depend on glutathione-S-transferases (GSTs). Similarly, Enayati et al. (2005) also stressed the importance of GST in SP acaricide metabolism. Previous studies have documented the involvement of GST in the breakdown of both endogenous and exogenous substances through either an elevated expression of the GST gene or an increase in enzyme activity (Hernandez et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe current findings corroborate the previously reported data on increased level of esterase of resistant \u003cem\u003eR. microplus\u003c/em\u003e ticks procured from Haryana, as shown by Gaur et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In a similar vein, the research carried out by Ghosh et al. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) demonstrated a noteworthy escalation in the levels of esterase, monooxygenase, and GST activity in the resistant \u003cem\u003eR. microplus\u003c/em\u003e isolates obtained from Bihar in Eastern India. In another study, Kumar et al. (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) noted the significantly higher esterase activity in the tick populations sampled from different states of India compared to the reference IVRI-I strain ranging from 0.096\u0026thinsp;\u0026plusmn;\u0026thinsp;0.045 to 7.893\u0026thinsp;\u0026plusmn;\u0026thinsp;0.302.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe resistance development status of ticks procured from eight districts of Marathwada region of Maharashtra clearly indicates that the SP compounds (deltamethrin and cypermethrin) are not working efficiently for tick control. Ticks infested animals in Marathwada region responded differently to these acaricides, and so region-specific control strategies in such a large state need to be worked out. The use of SP compounds should be regulated in this region to prevent further establishment of resistant populations and other newer drugs for tick management should be introduced. Besides, more studies on acaricide resistance in other divisions of the state needs to be carried out to work out present status of tick resistance. Effective management of the current situation and the preservation of a secure environment require collaborative efforts including all stakeholders.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contribution\u0026nbsp;\u003c/strong\u003eConceptualisation: GC, NJ. Data curation: AS. Formal analysis: SK. Investigation: SK, MV, SKu, MFMFS. Methodology: GC, AD. Supervision: GC, AS. Writing\u0026mdash;original draft: SK, MR. Writing \u0026mdash; review and editing: AKS, MR, GC\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e Not applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e Not applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e Not applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e The authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding \u0026nbsp;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to Associate Dean, College of Veterinary \u0026amp; Animal Sciences, MAFSU, Parbhani for providing facilities to carry out research work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbbas RZ, Zaman MA, Colwell DD, Gilleard J, Iqbal Z (2014) Acaricide resistance in cattle ticks and approaches to its management: the state of play. Vet Parasitol 203(1\u0026ndash;2): 6\u0026ndash;20.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.vetpar.2014.03.006\u003c/span\u003e\u003cspan address=\"10.1016/j.vetpar.2014.03.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAhanger RR, Godara R, Katoch R, Yadav A, Bhutyal ADS, Katoch M, Bader MA (2015) Deltamethrin resistance in field populations of \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e (Acari: Ixodidae) in Jammu and Kashmir, India.Exp Appl Acarol 67:467\u0026ndash;475.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlonso-Diaz MA, Rodriguez-Vivas RI, Fragoso-Sanchez H, Rosario-Cruz R (2006) Ixodicide resistance of the the \u003cem\u003eBoophilus microplus\u003c/em\u003e tick to ixodicides. Archivos de medicina veterinaria 38(2):105\u0026ndash;113.DOI:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4067/S0301-732X2006000200003\u003c/span\u003e\u003cspan address=\"10.4067/S0301-732X2006000200003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaffi MA, De Souza GRL, De Sousa CS, Ceron CR, Bonetti AM (2008) Esterase enzymes involved in pyrethroid and organophosphate resistance in a Brazilian population of \u003cem\u003eRiphicephallus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e (Acari, Ixodidae). Mol Biochem Parasitol 160(1):70\u0026ndash;73. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.molbiopara.2008.03.009\u003c/span\u003e\u003cspan address=\"10.1016/j.molbiopara.2008.03.009\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBradford M M (1976) A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principles of protein dye binding. Anal Biochem 72: 248\u0026ndash;254.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChigure GM, Sharma AK, Kumar S, Fular A, Sagar SV, Nagar G, Ghosh S (2018) Role of metabolic enzymes in conferring resistance to synthetic pyrethroids, organophosphates, and phenylpyrazole compounds in \u003cem\u003eRhipicephalus microplus\u003c/em\u003e. Int J Acarol 44(1):28\u0026ndash;34. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/01647954.2017.1400588\u003c/span\u003e\u003cspan address=\"10.1080/01647954.2017.1400588\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ede Castro JJ (1997) Sustainable tick and tickborne disease control in livestock improvement in developing countries. Vet Parasitol 71(2\u0026ndash;3):77\u0026ndash;97.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0304-4017(97)00033-2\u003c/span\u003e\u003cspan address=\"10.1016/S0304-4017(97)00033-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Le\u0026oacute;n AAP, Mitchell RD III, Watson DW (2020) Ectoparasites of cattle. Vet Clin North Am Food Anim Pract 36(1):173\u0026ndash;185. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cvfa.2019.12.004\u003c/span\u003e\u003cspan address=\"10.1016/j.cvfa.2019.12.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFAO (1971) Recommended methods for the detection and measurement of resistance of agricultural pests to pesticides \u0026ndash; tentative method for larvae of cattle tick, \u003cem\u003eBoophilus microplus spp\u003c/em\u003e. FAO method No. 7. FAO Plant Protect. Bull. 19: 15\u0026ndash;18.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFAO (2004) Resistance Management and Integrated Parasite Control in Ruminants: Guidelines. Module 1. Ticks: Acaricide Resistance: Diagnosis, Management and Prevention. Animal Production and Health Division, Rome. FAO: pp. 25\u0026ndash;77.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFinny DJ (1962) Probit Analysis- A Statistical Treatment of the Response Curve. Cambridge University Press, Cambridgepp. 1\u0026ndash;318.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFular A, Gupta S, Sharma AK, Kumar S, Upadhaya D, Shakya M, Ghosh S (2020) Standardization of tick specific biochemical tools for estimation of esterases, monooxygenases and glutathione S-transferase for characterization of acaricide resistance. Pestic Biochem Phy 164:130\u0026ndash;139.DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.pestbp.2020.01.008\u003c/span\u003e\u003cspan address=\"10.1016/j.pestbp.2020.01.008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFular A, Sharma AK, Kumar S, Nagar G, Chigure G, Ray DD, Ghosh S (2018) Establishment of a multi-acaricide resistant reference tick strain (IVRI-V) of \u003cem\u003eRhipicephalus microplus\u003c/em\u003e. Ticks Tick Borne Dis9(5):1184\u0026ndash;1191.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ttbdis.2018.04.014\u003c/span\u003e\u003cspan address=\"10.1016/j.ttbdis.2018.04.014\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGaur RS, Sangwan AK, Sangwan N, Kumar S (2016) Acaricide resistance in \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e and \u003cem\u003eHyalommaanatolicum\u003c/em\u003e collected from Haryana and Rajasthan states of India. Exp Appl Acarol69:487\u0026ndash;500. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10493-016-0046-1\u003c/span\u003e\u003cspan address=\"10.1007/s10493-016-0046-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh S, Azhahianambi P, De la Fuente J (2006) Control of ticks of ruminants, with special emphasis on livestock farming systems in India: present and future possibilities for integrated control\u0026mdash;a review. Exp Appl Acarol 40:49\u0026ndash;66. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10493-006-9022-5\u003c/span\u003e\u003cspan address=\"10.1007/s10493-006-9022-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh S, Bansal GC, Gupta SC, Ray D, Khan MQ, Irshad H, Ahmed JS (2007) Status of tick distribution in Bangladesh, India and Pakistan. Parasitol Res 101:207\u0026ndash;216. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s00436-007-0684-7\u003c/span\u003e\u003cspan address=\"10.1007/s00436-007-0684-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh S, Gupta S, Kumar KA, Sharma AK, Kumar S, Nagar G, Ray D (2017) Characterization and establishment of a reference deltamethrin and cypermethrin resistant tick line (IVRI-IV) of \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e. Pestic Biochem Phys 138:66\u0026ndash;70. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.pestbp.2017.03.002\u003c/span\u003e\u003cspan address=\"10.1016/j.pestbp.2017.03.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh S, Kumar R, Nagar G, Kumar S, Sharma AK, Srivastava A, Saravanan BC (2015) Survey of acaricides resistance status of \u003cem\u003eRhipiciphalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e collected from selected places of Bihar, an eastern state of India. Ticks Tick Borne Dis 6(5):668\u0026ndash;675. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ttbdis.2015.05.013\u003c/span\u003e\u003cspan address=\"10.1016/j.ttbdis.2015.05.013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh SS, Azhahianambi PA, Yadav MP (2007) Upcoming and future strategies of tick control: a review. J Vect Borne Dis 44(2):79\u0026ndash;89\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGodara R, Katoch R, Rafiqi SI, Yadav A, Nazim K, Sharma R, Katoch M (2019) Synthetic pyrethroid resistance in \u003cem\u003eRhipicephalus (Boophilus) microplus\u003c/em\u003e ticks from north-western Himalayas, India. Trop Anim Health Prod 51:1203\u0026ndash;1208. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11250-019-01810-8\u003c/span\u003e\u003cspan address=\"10.1007/s11250-019-01810-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGraf JF, Gogolewski R, Leach-Bing N, Sabatini GA, Molento MB, Bordin EL, Arantes GJ (2004) Tick control: an industry point of view. Parasitol 129(S1):S427-S442. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0031182004006079\u003c/span\u003e\u003cspan address=\"10.1017/S0031182004006079\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuerrero F, P\u0026eacute;rez de Le\u0026oacute;n A, Rodriguez-Vivas R, Jonsson N, Miller R, Andreotti R. E. N. A. T. O, Roe RM (2014) Acaricide research and development, resistance and resistance monitoring. Sonenshine DE, Roe RM, editors, 2:353\u0026ndash;374.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGupta S, Sangwan N, Sangwan AK, Mann S, Gupta S, Kumar A, Kumar S (2023) Understanding the resistance mechanisms of \u003cem\u003eRhipicephalus microplus\u003c/em\u003e ticks to synthetic pyrethroids and organophosphates in south-west regions of Haryana, North India. Pestic Biochem Physiol 196: 105634. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.pestbp.2023.105634\u003c/span\u003e\u003cspan address=\"10.1016/j.pestbp.2023.105634\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHemingway J, Hawkes NJ, McCarroll L, Ranson H (2004) The molecular basis of insecticide resistance in mosquitoes. Insect Biochem Mol Biol 34(7):653\u0026ndash;665.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ibmb.2004.03.018\u003c/span\u003e\u003cspan address=\"10.1016/j.ibmb.2004.03.018\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHernandez EP, Kusakisako K, Talactac MR, Galay RL, Hatta T, Fujisaki K, Tanaka T (2018) Glutathione S-transferases play a role in the detoxification of flumethrin and chlorpyrifos in \u003cem\u003eHaemaphysalis longicornis\u003c/em\u003e.Parasit Vectors 11:1\u0026ndash;14.DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s13071-018-3044-9\u003c/span\u003e\u003cspan address=\"10.1186/s13071-018-3044-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJonsson NN, Mayer DG, Matschoss AL, Green PE, Ansell J (1998) Production effects of cattle tick (\u003cem\u003eBoophilus microplus\u003c/em\u003e) infestation of high yielding dairy cows. Vet Parasitol 78(1):65\u0026ndash;77.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0304-4017(98)00118-6\u003c/span\u003e\u003cspan address=\"10.1016/S0304-4017(98)00118-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRinesh Kumar, Nagar G, Sharma AK, Kumar S, Ray DD, Chaudhuri P, Ghosh S (2013) Survey of pyrethroids resistance in Indian isolates of \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e: identification of C190A mutation in the domain II of the para-sodium channel gene. Acta Trop 125(2): 237\u0026ndash;245. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.actatropica.2012.10.006\u003c/span\u003e\u003cspan address=\"10.1016/j.actatropica.2012.10.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar R, Sharma AK, Ghosh S (2020) Menace of acaricide resistance in cattle tick, \u003cem\u003eRhipicephalus microplus\u003c/em\u003e in India: Status and possible mitigation strategies. Vet Parasitol 278, 108993. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.vetpar.2019.108993\u003c/span\u003e\u003cspan address=\"10.1016/j.vetpar.2019.108993\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar S, Sharma AK, Kumar B, Shakya M, Patel JA, Kumar B, Ghosh S (2021) Characterization of deltamethrin, cypermethrin, coumaphos and ivermectin resistance in populations of \u003cem\u003eRhipicephalus microplus\u003c/em\u003e in India and efficacy of an antitick natural formulation prepared from \u003cem\u003eAgeratum conyzoides\u003c/em\u003e. Ticks Tick Borne Dis 12(6):101818.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ttbdis.2021.101818\u003c/span\u003e\u003cspan address=\"10.1016/j.ttbdis.2021.101818\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar SS, Rayulu VC, Rao KS, Kumar NV (2017) Acaricidal resistance in \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003eMicroplus\u003c/em\u003e ticks infesting cattle of Andhra Pradesh. J Entomol Zool Stud 5(6):580\u0026ndash;584.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaske DK, Sardey MR, Bhilegaonkar NG (1997). Prevalence of ixodid ticks on cattle in Vidarbha region of Maharashtra. Indian I Anim Sci67(9): 768\u0026ndash;769.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiller TA (1988) Mechanisms of resistance to pyrethroid insecticides. Parasitology Today 4(7):S8-S12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003eDOI:https://doi.org/10.1016/0169-4758(88)90080-4\u003c/span\u003e\u003cspan address=\"DOI:10.1016/0169-4758(88)90080-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMinjauw B, McLeod A (2003) Tick-borne diseases and poverty: the impact of ticks and tick-borne diseases on the livelihoods of small-scale and marginal livestock owners in India and eastern and southern Africa. Tick-borne diseases and poverty: the impact of ticks and tick-borne diseases on the livelihoods of small-scale and marginal livestock owners in India and eastern and southern Africa.124-pp\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh O, Bhardwaj P, Singh J (2017) Distribution of Lightning Casualties over Maharashtra, India. J Ind Geophys Union 21(5): 415\u0026ndash;424\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRobertson JL,Russell RM, Priesler HK,Savin NE (2007) Bioassay with arthropods, second edn. CRC Press, Boca Raton.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1201/9781420004045\u003c/span\u003e\u003cspan address=\"10.1201/9781420004045\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRodrigues DS, Leite RC (2013) Economic impact of \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e: estimate of decreased milk production on a dairy farm. Arquivo Brasileiro de Medicina Veterin\u0026aacute;ria e Zootecnia, 65:1570\u0026ndash;1572.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRosario-Cruz R, Almazan C, Miller R, Dominguez-Garcia DI, Hernandez-Ortiz R (2009) Front Biosci (Landmark Ed) 14(7): 2657\u0026ndash;65. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2741/3403\u003c/span\u003e\u003cspan address=\"10.2741/3403\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShakya M, Singh M, Kumar S, Jayraw AK, Jatav GP, Agrawal V, Jamara N (2021) Efficacy of commercially available chemical compound used against management of tick infestation in Mhow, Madhya Pradesh. Int J Trop Insect Sci 41:2469\u0026ndash;2475. DOI:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s42690-020-00424-0\u003c/span\u003e\u003cspan address=\"10.1007/s42690-020-00424-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma AK, Kumar R, Kumar S, Nagar G, Singh NK, Rawat SS, Ghosh S (2012) Deltamethrin and cypermethrin resistance status of \u003cem\u003eRhipicephalus\u003c/em\u003e (\u003cem\u003eBoophilus\u003c/em\u003e) \u003cem\u003emicroplus\u003c/em\u003e collected from six agro-climatic regions of India. Vet Parasitol 188(3\u0026ndash;4):337\u0026ndash;345. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.vetpar.2012.03.050\u003c/span\u003e\u003cspan address=\"10.1016/j.vetpar.2012.03.050\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShaw RD (1966) Culture of an organophosphorus-resistant strain of \u003cem\u003eBoophilus microplus\u003c/em\u003e (Can.) and an assessment of its resistance spectrum. Bull Ento Res 56(3):389\u0026ndash;405. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0007485300056480\u003c/span\u003e\u003cspan address=\"10.1017/S0007485300056480\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShyma KP, Kumar S, Sharma AK, Ray DD, Ghosh S (2012) Acaricide resistance status in Indian isolates of \u003cem\u003eHyalomma anatolicum\u003c/em\u003e. Exp Appl Acarol 58:471\u0026ndash;481. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10493-012-9592-3\u003c/span\u003e\u003cspan address=\"10.1007/s10493-012-9592-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShyma KP, Singh V, Gupta JP (2019) Susceptibility Level of \u003cem\u003eOrnithodoros\u003c/em\u003e spp (Acari: Argasidae) to Some Commercial Synthetic Pyrethroids in North Gujarat. Int J Livest Res 9(3):62\u0026ndash;67. DOI:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5455/ijlr.20180522085604\u003c/span\u003e\u003cspan address=\"10.5455/ijlr.20180522085604\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShirsikar PM, Narladkar BW, Jadhav ND, Rajurkar SR, Jadhav SG, Channa GR, Gaikwad SS, Chigure GM (2023) Morphological characterization and prevalence of hard ticks infesting cattle of Marathwada Region, Maharashtra state, India. J Exp Zool India26(2). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.51470/jez.2023.26.2.2575\u003c/span\u003e\u003cspan address=\"10.51470/jez.2023.26.2.2575\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSindhu ZUD, Naseer MU, Raza A, Aslam B, Ahmad J, Abbas RZ, Khattak B (2022) Resistance to Cypermethrin Is Widespread in Cattle Ticks (\u003cem\u003eRhipicephalus microplus\u003c/em\u003e) in the Province of Punjab, Pakistan: In Vitro Diagnosis of Acaricide Resistance. Pathogens 11(11):1293. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pathogens11111293\u003c/span\u003e\u003cspan address=\"10.3390/pathogens11111293\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan Wyk RD, Baron S, Maritz-Olivier C (2016) An integrative approach to understanding pyrethroid resistance in \u003cem\u003eRhipicephalus microplus\u003c/em\u003e and \u003cem\u003eR\u003c/em\u003e. \u003cem\u003edecoloratus\u003c/em\u003e ticks. Ticks Tick Borne Dis 7(4):586\u0026ndash;594. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ttbdis.2016.01.007\u003c/span\u003e\u003cspan address=\"10.1016/j.ttbdis.2016.01.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"parasitology-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pare","sideBox":"Learn more about [Parasitology Research](http://link.springer.com/journal/436)","snPcode":"436","submissionUrl":"https://submission.nature.com/new-submission/436/3","title":"Parasitology Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Rhipicephalus microplus, Resistance, Deltamethrin, Cypermethrin, Metabolic enzymes, Maharashtra","lastPublishedDoi":"10.21203/rs.3.rs-4464602/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4464602/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study examined the pattern of resistance to widely applied synthetic pyrethroids i.e. cypermethrin and deltamethrin, against larvae of \u003cem\u003eRhipicephalus microplus\u003c/em\u003e ticks sampled from Marathwada region in Maharashtra, India. The study also examined the role of α and β-esterase\u0026rsquo;s and glutathione-S-transferase (GST) in resistance development. All eight \u003cem\u003eR. microplus\u003c/em\u003e isolates tested were resistant to deltamethrin (RL IV), having RR\u003csub\u003e50\u003c/sub\u003e values from 6.88 to 131.26. LPT analysis exhibited the resistance level II deltamethrin resistance in Beed and Hingoli, III in Dharashiv, and IV in Sambhajinagar, Parbhani, Latur, Jalna, and Nanded isolates. The LIT analysis showed that Dharashiv field isolates had the lowest LC\u003csub\u003e50\u003c/sub\u003e value of 229.09 ppm against cypermethrin, while Sambhajinagar field isolates had the highest at 489.78 ppm. The RR\u003csub\u003e50\u003c/sub\u003e ranged from 1145.45 to 2448.9. Seven isolates were level I resistant to cypermethrin while the Jalna isolate was level II resistant. In larvae treated with deltamethrin and cypermethrin, the activity of α and β-esterase enzymes increased significantly compared to control groups. The enzyme ratios in treated larvae ranged from 0.7533 to 1.7023 for α-esterase and 0.7434 to 3.2054 for β-esterase. The Hingoli isolate treated with cypermethrin exhibited the highest α-esterase activity (903.261), whereas, Sambhajinagar isolate had the highest GST enzyme ratio (2.8224) after deltamethrin exposure. When exposed to cypermethrin, the Hingoli isolate showed the highest GST enzyme ratio, 2.0832.\u003c/p\u003e","manuscriptTitle":"Current profile of pyrethroid resistance in Rhipicephalus microplus populations sampled from Marathwada region of Maharashtra state, India","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-10 06:29:40","doi":"10.21203/rs.3.rs-4464602/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"25240797634649840630996883815656267483","date":"2024-06-05T04:32:49+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"335275773748746888329181330619344291226","date":"2024-06-04T15:50:42+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"257656504328258396301184231645472928259","date":"2024-06-04T13:33:18+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-06-04T11:24:06+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-05-29T07:10:33+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-05-28T02:35:39+00:00","index":"","fulltext":""},{"type":"submitted","content":"Parasitology Research","date":"2024-05-23T06:19:14+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"parasitology-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pare","sideBox":"Learn more about [Parasitology Research](http://link.springer.com/journal/436)","snPcode":"436","submissionUrl":"https://submission.nature.com/new-submission/436/3","title":"Parasitology Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"53928c6e-d139-4ce5-a65f-3864f3896920","owner":[],"postedDate":"June 10th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-08-01T17:16:55+00:00","versionOfRecord":{"articleIdentity":"rs-4464602","link":"https://doi.org/10.1007/s00436-024-08306-x","journal":{"identity":"parasitology-research","isVorOnly":false,"title":"Parasitology Research"},"publishedOn":"2024-07-22 16:15:02","publishedOnDateReadable":"July 22nd, 2024"},"versionCreatedAt":"2024-06-10 06:29:40","video":"","vorDoi":"10.1007/s00436-024-08306-x","vorDoiUrl":"https://doi.org/10.1007/s00436-024-08306-x","workflowStages":[]},"version":"v1","identity":"rs-4464602","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4464602","identity":"rs-4464602","version":["v1"]},"buildId":"ehx78VzkSd0WSzXnipQa-","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}