Response of seven populations of the two-spotted spider mite (tetranychus urticae koch) for bifenazate acaricide on cucumber (cucumis sativus l.) under plastic houses in Jordan.
Two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae), is a polyphagous pest. It is a major pest on field crops, plastic-houses crops, horticultural crops, ornamentals and fruit trees [18,17,2]. It has recently become a serious problem because of the extensive use of acaricides, resulting in resistance among the mite populations [5,15]. The development of the resistance is also known to be accelerated under confined environmental conditions such as plastic-houses . Since the mite has a very short life cycle, its resistance to acaricides has more readily emerged than the rate of the resistance in other pests. In addition, the mites resistance to certain acaricides has been shown to have cross resistance to other acaricides. Thus, most commercial acaricides have been often proved to be ineffective to control the field mite populations [13,38].
From field surveillance and screening of various acaricides, it was speculated that TSSM has developed resistance to the most conventional acaricides, but toxicological data are very scarce and poorly documented . It is quite possible that TSSM susceptibility to acaricides would differ from one location to another of cucumber cultivation in Jordan. Therefore, it was important to monitor the acaricide susceptibilities of T. urticae populations that were collected from cucumber cultivation in Jordan and to evaluate the efficacy of testing acaricides. This study showed the results of laboratory-based tests that determined the response of six field populations (Al-Ramtha, Baq'a, Zyzya, Krimeh, Deir-Alla, and Karamah) and one susceptible strain of T. urticae to bifenazate acaricide.
In 2009, the total number of plastic-houses in Jordan, which were planted with vegetables, was about 66,000. More than fifty percent of them were planted with cucumber (36,000). The other fifty percent were planted with tomato, pepper, eggplant, beans and other crops. .
The Jordanian farmers rely heavily on acaricides to control the two-spotted spider mite. Therefore, they have increased the rate of application, applied a mixture of acaricides and applied acaricides more frequently than they should. They have complained about unsatisfactory results in controlling T. urticae . The total quantity of acaricides imported to Jordan in 2009 was 120,000 liters and / or kilograms. The imported acaricides belong to 18 active ingredients. Of these bifenazate recently used .
Although TSSM represents a real threat to cucumber plantation under plastic-houses in Jordan, few toxicological studies on this pest have been conducted . Therefore, this study aimed to achieve the following objectives: to evaluate the susceptibilities of TSSM collected from cucumber cultivation in Jordan to the studied bifenazate and to find the resistance factors to the tested acaricide, bifenzate among the two spotted spider mite (TSSM) populations.
Materials And Methods
Populations of T. urticae:
Six T. urticae populations were collected from cucumber plants grown under plastic houses conditions in different regions of Jordan. These geographical regions include Al-Ramtha (100 Km North West of Amman), Baq'a (20 Km north west of Amman), Zyzya (30 Km South of Amman), Krimeh (South of Jordan Valley), Deir-Alla (Central of Jordan Valley) and Karamah (South of Jordan Valley). These regions are considered the main area for cucumber production in the country. A susceptible strain of T. urticae was obtained from Lattakia Center for Rearing and Production of Biological Agents (LCRPBA) in Syria. This strain was reared in (LCRPBA) for 5 years without the application of acaricides.
Production of bean plants:
Bean (Phaseolus vulgaris L. cv. Bronco) was chosen for rearing and for the toxicological tests of the (TSSM) because it is one of the mite's favorite host, and for its ease of producing transplants. Seeds of bean (Bronco, Asgrow, USA) were directly sowed inside 10 cm. pots. Potting media used was Peatmoss and Perlite with 3:1 ratio. Plants were infested with SSS T. urticae when they reached the true leaf stage. These plants were irrigated and replaced as needed. No pesticides were applied on the plants except for acaricide tests. These plants were grown under greenhouse conditions.
For use in the toxicological tests, polystyrene trays of 84 cells were filled by Peat-moss and Perlite (3:1 ratio). Then, these trays were sown by 1-2 bean seeds for each cell, after complete germination the cotyledon leaves were used.
Rearing of the Syrian susceptible strain)
Rearing of the Syrian susceptible strain:
(SSS) was done inside special insectaria in University of Jordan at Al Jubaiha area. The Syrian TSSM was reared and maintained on Phaseolus vulgaris plants under greenhouse conditions. P. vulgaris Plants were irrigated and replaced as needed.
P. vulgaris seeds were sowed in 84 polystyrene trays filled with Peat-moss and Perlite at ratio of 3:1. Each cell was sowed with one or two seeds. Prior to germination, cotyledon leaves of seedling were used for all experimental sets.
Bifenazate 24% W/V, SC (Floramite(r)). Produced by Uniroyal Chemical Co., Inc. (now part of Crompton Corp.) in 2000, with higher recommended rate of 96 mg/LH2O. Its Molecular formula is C17H20N2O3 .
Preparing of stock solutions:
For each test, fresh stock solution was prepared by dissolving a calculated quantity of the acaricide enough to run the whole concentrations needed. The acaricide was used as its commercial formulation (Floramite 24% SC).
Bioassay of the acaricide toxicity:
Toxicological bioassays were conducted according to the procedures described by IRAC . Cotyledon leaves from untreated bean plants were placed, lower side up, in Petri-dishes lined with water-saturated cotton wool. 25 adult females of T. urticae were introduced into each cotyledon leaf by using a binocular microscope and a fine paintbrush. 24 hours after mite release, each Petri-dish was sprayed with a constant amount of the acaricide solutions for 2 seconds using a hand sprayer. The sprayed Petri-dishes were left to dry for 30 minutes, then they were placed under room temperature. Mites condition was assessed by gentle probing with a fine paintbrush. Mites were classed as dead when they didn't move or displayed some movement not exceeding their body length. Mites which were able to move were considered alive. This assessment of mites conditions were recorded 48 hours after acaricide application. The LC50, and LC90 values and their 95% confidence limits were calculated from probit regressions using the SPSS13 program [14,20].
The application of the acaricide was done with four replicates per each concentration, and with seven concentrations for the acaricide. These concentrations were chosen based on preliminary studies and they were different for each T. urticae population. Tap water was sprayed as control. These experiments were carried out in the Pesticide laboratory at temperature between 25,5 C and 57,8% relative humidity.
Data were subject to probit analysis  which incorporated Abbott's correction for natural mortality . The SPSS (version 13 USA) computer program was used for data analysis to estimate LC50 and LC90 values, regression coefficient (slope) and its standard error, intercept and its standard error, Pearson goodness of fit chi-square, expected mortality, and 95% confidence limits (95%CL) for effective level of concentrations. This programme used normal equivalent deviate (NED) instead of probit numbers. However, NED numbers can be readily adjusted to probit by adding 5 to each NED number (Finney, 1971). Y value for each line estimated by probit regression was equal to 0.0 and 1.28 when LC50 and LC90 (X) value was converted to log base 10, respectively.
LC50 and LC90 values were considered significant when (95% CL) did not overlap. To determine the resistance factor (RF) for each population, the LC50 of each acaricide of the field population was divided by the corresponding LC50 for the susceptible strain. The resistance factors were categorized according to Fukami (1983) as follows: low. RF<10, moderate 10<RF[less than or equal to]40, high 40-60 and very high resistance >60. LC90 values in ppm divided by the higher recommended field rate in ppm were calculated and tabulated for each TSSM strain (Ratio value). Goodness of line fitting was checked by Chi-square test X2. According to Finney (1971), the value of X2 at 0.05 level of probability equals to 14.1 at 5 degree of freedom (df). Results obtained in this study revealed that (X2) Goodness of fit chi-square were less than that tabulated for each regression line indicating goodness of fit at 0.05 level of probability.
Based on resistance factor values, Deir-Alla (RF=1.72) and Krimeh (RF=1.14) populations seem to initiate resistance development to bifenazate with RF of 1.7 and 1.1, respectively. However, Populations from Al-Ramtha with (RF=1.0), Zyzya with (RF=0.86), Karamah with (RF=0.58), Baq'a with (RF=0.53) were susceptible to bifenazate (Table1). LC50 for Baq'a, Karamah, and Zyzya populations were 12.13, 13.90, 19.73ppm, respectively which were significantly susceptible to bifenazate compared to SSS population with 23.06 ppm. Populations from Al-Ramtha region (LC50=23.1 ppm) showed almost equal susceptibility to bifenazate compared to SSS strain, but LC50 of 39.93 ppm for Deir-Alla population was significantly less susceptible than SSS strain.
However, there were no significant differences between LC50 values for the populations from Karamah, and Baq'a, as well as LC50 values were not significantly different for the populations from Zyzya, Al-Ramtha and the SSS strain, while LC50 value for Deir-Alla population was significantly different from all the other populations.
The ratio between LC90 and higher recommended field rate indicated that Baq'a population was the most susceptible to bifenazate toxicity (Ratio =0.37) followed by Karamah (0.38), Al-Ramtha (0.66), Krimeh (0.74), Zyzya (0.84) and Deir-Alla (1.07) populations (Table 1).
Monitoring of local populations for susceptibility towards acaricides is the first step in resistance management of T. urticae. It is essential to carry out acaricide resistance tests regularly to avoid resistance development in target mites. In addition, control tactics must depend on the use of different acaricides to avoid or delay resistance.
The toxicity of bifenazate on one foreign susceptible mite strain obtained from Syria and six local populations collected from different locations revealed that two of local populations of T. urticae (Deir-allon and krimeh) were resistance to bifenazate whereas the other four tested populations and foreign population was susceptible. Resistance to bifenazate could be because of the frequent use of bifenazate by Jordanian farmers in a large scale. Since Deir-Allah and krimeh region are mainly cultivated with eggplant throughout the year which is a favorite host for T urticae, farmers use acaricides including bifenazate in a large scale. Therefore, mite populations originated from this location exhibited the highest resistance to bifenazate. In agreement with this result, Lee et al.,  pointed out that the resistance factors of some acaricides particularly abamectin varied between 0.3 to 19.5 towards T. urticae eight populations collected from different locations in Korea. Bifenazate was found to provide excellent control against adult stages of T. urticae at low concentrations. Deir-Alla population was the most resistant population with 1.7 resistance factor, while other populations were nearly susceptible to bifenazate. Explanations for that can be rely on the fact that bifenazate is anew acaricide, it was registered in Jordan in 2005, it has different mode of action in comparison with other used acaricides and its cost is high compared with abamectin and amitraz In Japan, Ochiai et al.  concluded that bifenazate remained effective on controlling T. urticae adult females. Their estimated LC50 for bifenazate against T. urticae was 0.63 ppm. The authors reported that bifenazate provided excellent control against adult and larval stages of T. urticae at low concentrations but was less effective on the egg stage. In Belgium, Van Leeuwen et al.,  reported that bifenazate-resistant strain lacked cross resistance to many different chemical classes and modes of action of other acaricides. In South Korea, Lee et al.  concluded that all T. urticae populations tested had low to moderate resistance to bifenazate. Resistance factors ranged from 0.8 to 11.0. The same authors mentioned that the resistance to other acaricides might result in the cross resistance to bifenazate. However, Martinson et al. (1991) concluded that the reference susceptible population, and the bioassay methods differ among researchers, and it is therefore not easy to compare these results.
In couclusion, two of the tested field populations Deir-Allah and krimeh were resistant to bifenazate. The resistance factor ranged from 0.58 to 1.73. At its high recommended field rate bifenazate was effective in controlling T. urticae. It had good residual effect for all tested population except against the Deir-Allah and krimeh populations. However, Bifenazate was a very useful acaricide giving a high efficacy, long lasting effect against T. urticae even when it was used with concentrations less than its recommended application rate. However, In order to have safe and high quality and quantity of cucumber product and to manage resistance development by T. urticae to acaricides, the following are recommended., Regular monitoring should be carried out to detect the extent of resistance to the pesticides used in Jordan or every where, particularly bifenazate, restricting the use of acaricides to which the magnitude of resistance is high, establishing of baseline LC50 to new acaricide before widespread use which allow better monitoring of changes in susceptibility over time. applying acaricides that have different active ingredients and different mode of action., training of growers on alteration of acaricides based on mode of action to facilitate long term sustainable spider mite management for agriculture in Jordan and studying the mechanisms of resistance to acaricides is very important to prevent cross resistance between closely related groups.
Thanks are extended to the Deanship of research at university of Jordan and Agriculture Materials company administration for finance support. Thanks are also to the staff at the lattakia (enter for rearing and production of biological Agents in Syria for providing us the sensitive strain of the spider mite.
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(1) Tawfiq M.Al-Antary, (2) Mohammad Raed kame l Al Lala and (3) Marwan I. Abdel-Wali
(1) Plant Protection Dept, Faculty of Agriculture, University of Jordan, Amman, Jordan.
(2) Ph.D student
(3) Pesticide Toxicology, Ph.D, National Center of Agriculture Research and Extension, Ministry of Agriculture, Al-Baqae, Jordan.
Tawfiq M. Al-Antary, Plant Protection Dept, Faculty of Agriculture, University of Jordan, Amman, Jordan. E-mail: firstname.lastname@example.org
Table 1: Susceptibilities of field-collected populations of T. urticae adult females to bifenazate (Higher recommended field rate = 96 mg/L). Population LC50(mg/l) LC90(mg/l) L.E.P.R2 name 95%CL1 95%CL Y=a+b(x) Ar-Ramtha 23.10 bc6 62.97b6 Y=-4.01+2.94(x) 20.31-25.62 55.11-75.32 Baq'a 12.13 d 35.52c Y=-2.98+2.75(x) 10.74-13.44 30.91-42.63 Zyzya 19.73 c 80.79ab Y=-2.71+2.09(x) 17.02-22.41 67.12-102.90 Krimeh 26.21 b 70.71b Y=-4.22+2.97(x) 23.51-28.77 62.16-83.66 Deir-Alla 39.93 a 102.96a Y=-4.99+3.12(X) 35.94-43.67 91.12-120.62 Karamah 13.36 d 36.38c Y=-3.32+2.95(x) 11.99-14.66 31.91-43.20 SSS 23.06 bc 63.70b Y=-3.96+2.90(x) 20.50-25.41 56.23-74.66 Population Slope [+ or -] Ratio4 RF5 Chi Square name S.E3. LC90 LC50 Calculated Ar-Ramtha 2.94 [+ or -] 0.66 1.0 4.3 0.28 Baq'a 2.75 [+ or -] 0.37 0.53 5.5 0.23 Zyzya 2.09 [+ or -] 0.84 0.86 7.4 0.17 Krimeh 2.97 [+ or -] 0.74 1.14 7.7 0.25 Deir-Alla 3.12 [+ or -] 1.07 1.73 6.3 0.26 Karamah 2.95 [+ or -] 0.38 0.58 5.2 0.25 SSS 2.90 [+ or -] 0.66 -- 3.6 0.24 95% Confidence limits for LC50 or LC90 in ppm. L.E.P.R. = Line Estimated by Probit Regression. S.E. = Standard error. Ratio LC90 = LC90 / higher recommended field rate. R.F. Resistance Factor = LC50 of field population / LC50 of susceptible population (SSS). LC50 or LC90 values having different letters are significantly different (95% CL did not overlap).
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|Title Annotation:||Original Article|
|Author:||Antary, Tawfiq M. Al-; Lala, Mohammad Raed kame l Al; Abdel-Wali, Marwan I.|
|Publication:||Advances in Environmental Biology|
|Date:||Jun 1, 2012|
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