TY - JOUR AU - Gondim, Manoel G, C AB - Abstract Walking is important to dispersal on plants and colonization of new plants by predatory mites, and this activity is potentially affected by the presence of acaricides. This possibility was investigated in coconut fruits infested with the coconut mite, Aceria guerreronis Keifer (Acari: Eriophyidae), where colonization by the predator Neoseiulus baraki (Athias-Henriot) (Acari: Phytoseiidae) was monitored. The following acaricides were evaluated for influence on the process of colonization by the predatory mite: abamectin, azadirachtin, and fenpyroximate. Water-treated fruits were used for comparison. Experiments were conducted with and without freedom of choice on coconut fruits with the release and recapture of females of N. baraki marked with fluorescent ink. A confinement experiment was also carried out on coconut bunch rachis sprayed or not sprayed with the acaricides. The predatory mite N. baraki avoided contact with acaricide-contaminated areas. After the predators were released on the fruits or bunch rachis, larger numbers were recaptured under the bracts than on the surface of the fruits. The number of predators recaptured in all experiments was lower in the treatments with acaricides than in the control. Among the acaricides tested, azadirachtin least affected N. baraki colonization. Therefore, the presence of the tested acaricides indeed interferes with N. baraki dispersal within plants and the coconut fruit colonization. intraplant dispersal, recapture, predator, walking activity, biocontrol agent Dispersal is an important behavior comprising distinct steps related to departure from a location (emigration), movement between habitats (displacement), and establishment elsewhere (immigration) (Begon et al. 2007, Ronce et al. 2007, Clobert et al. 2009). Through dispersal, the exploration of various environmental resources in time and space is possible (Ronce et al. 2007), which makes the behavior essential for survival (Safriel and Ritte 1980), with consequences for population dynamics, spatial distribution, and population genetics (Bowler and Benton 2005). The dispersal of mites may be either passive or active. Passive dispersal usually occurs when mites are transported on plants and plant parts by anthropic action (Evans 1992). Mites may also adhere to the bodies of vertebrate and invertebrate animals that carry them involuntarily (phoresy) (Perotti and Braig 2009) or may be transported by air currents, which can move them longer distances (Sabelis and Afman 1994, Tixier et al. 1998, Monteiro et al. 2018). Actively, mites can disperse by walking, and because of their small size and absence of wings, they normally travel short distances (Auger et al. 1999, Strong et al. 1999, Melo et al. 2014). Predatory mites of the family Phytoseiidae perform active dispersal by walking between parts of the same plant or between neighboring plants (Berry and Hotzer 1990, Auger et al. 1999, Strong et al. 1999, Croft and Jung 2001, Lopez et al. 2017). This dispersal can begin during a food shortage or when seeking sites for oviposition and shelter (Sabelis and Dicke 1985, Croft and Jung 2001). This type of dispersal allows the predatory mite to find prey on the plant (Sabelis and Dicke 1985). However, dispersal may be affected by abiotic and biotic factors (Berry and Holtzer 1990) such as temperature, relative air humidity, light intensity, food scarcity, and mite age (Berry and Holtzer 1990, Zemek and Nachman 1998, Auger et al. 1999, Ghazy et al. 2016). The presence of environmental contaminants such as pesticides can also interfere with the interspecific relations of arthropods on plants (Cordeiro et al. 2014), as these products affect not only the target species but also beneficial species such as natural enemies and other nontarget species (Guedes et al. 2016, 2017). Pesticides affect the physiology, biology, and behavior of arthropods (Ibrahin and Yee 2000, Lima et al. 2015b, 2016), so they can influence the dispersal process (Desneux et al. 2007, Guedes et al. 2016, Monteiro et al. 2018). Behavioral avoidance to pesticides may take place by means of repellence and irritability (Cordeiro et al. 2010, Guedes et al. 2016). Repellence takes place when the arthropod is able to detect the presence of toxic substances (pesticides) in the environment and avoid direct contact with the contaminated area and disperse. Alternatively, irritability may take place when the dispersal occurs after contact with the toxic substances, favoring movement to areas free of these substances (Roberts et al. 1997, Cordeiro et al. 2010, Guedes et al. 2016). Therefore, pesticide-induced stress on the behavior of arthropods can affect their dispersal and population dynamics and directly or indirectly influence pest management (Guedes et al. 2016). This study explored the effect of acaricide residues used to control the coconut mite Aceria guerreronis Keifer (Acari: Eriophyidae) on fruit colonization by the predatory mite Neoseiulus baraki (Athias-Henriot) (Acari: Phytoseiidae). The predatory mite N. baraki is one of the natural enemies of A. guerreronis in America, Africa, and parts of Asia (Moraes et al. 2004, Lawson-Balagbo et al. 2008, Fernando et al. 2010, Negloh et al. 2011). These mites inhabit the perianth of the coconut fruit, a region covered by the bracts (Aratchige et al. 2007, Lawson-Balagbo et al. 2008, Reis et al. 2008). After colonizing the fruits, the population of A. guerreronis is reduced mainly by intraspecific competition, reduction in food quality, and predation (interspecific competition). Subsequently, the population of N. baraki also declines, with the mites dispersing due to a lack of food and intraspecific competition (Galvão et al. 2011). This dispersal occurs by the mites walking on the same plant in search of suitable feeding sites. However, these plants often exhibit acaricide residues applied to control the coconut mite A. guerreronis, which likely acts at the onset of dispersal (Melo et al. 2012, Monteiro et al. 2012), since acaricides do not affect mites that are under the bracts (Silva et al. 2017). To determine whether acaricides affect the colonization of N. baraki in fruits infested by A. guerreronis, the following aspects were determined: 1) whether the predator, when colonizing fruits infested with A. guerreronis, prefers those with or those without acaricide residue (i.e., whether the presence of the acaricide interfere in choice of the fruits by colonizing predator); 2) which acaricide least affects the colonization of fruits infested with A. guerreronis; and 3) whether N. baraki colonization depends on the location on the plant (and prey presence). Material and Methods Acquisition and Rearing of N. baraki Coconut fruits infested with A. guerreronis were collected on the island of Itamaracá, state of Pernambuco (PE), Brazil (07°46′S, 34°52′W), transported to the Laboratory of Acarology of the Federal Rural University of Pernambuco (UFRPE) in styrofoam boxes, and stored under refrigeration (approximately 10°C) for up to 5 d to obtain the mites. The fruit bracts were removed by pruning the shears and inserting a staple between the coconut surface and its bract allowing easy removal of the latter. The predatory (phytoseiid) mites were collected and mounted on slides with Hoyer’s medium and placed in an oven at 55°C. Next, the collected mites were observed on an Olympus BX41 optical microscope (Olympus Corporation, Tokyo, Japan) for identification (Moraes et al. 2004). After the species N. baraki was confirmed, the fruits were screened to leave approximately 100 individuals for each rearing unit. The rearing unit, approximately 16 cm in diameter, was made of a 1-mm-thick black polypropylene (PVC) film, filter paper, 1-cm-thick polyethylene foam, and a Petri dish, placed in this order. Hydrophilic cotton moistened with distilled water was placed on the edge of the PVC to prevent mite escape. Five fragments (approximately 1 cm3) of the perianth epidermis of coconut fruit infested with A. guerreronis (~80–100 individuals of different stages) were provided daily as food. The rearing units were kept in an incubator at 27 ± 0.5°C, 75 ± 5% relative humidity (RH), and 12 h photoperiod. Intraplant Release and Recapture of N. baraki Release of N. baraki on the Fruit Spikelet The experimental units were coconut fruits aged 3 to 4 mo (leaf 13 to 14), according to the coconut phyllotaxis (Sobral 1994), and showing 16 to 32% damage intensity (necrosis) by A. guerreronis according to the scale described by Galvão et al. (2008). Each infested fruit was fixed in a gypsum base molded in disposable plastic 200 ml cups, which were filled with 150 ml of gypsum diluted in water. A 6-cm-long nail was attached to the top of the base, with the plaster still fresh. After the plaster was dry, the plastic cup was removed. Each fruit was fixed on a gypsum base, with the nail inserted approximately 2 cm into the distal part of the fruit. In the region of contact of the fruit with the nail, a barrier was established with entomological glue (Isca Pega, Isca Ferramentas e Soluções para Manejo de Pragas), according to the methodology described by Silva et al. (2017). The spikelet of each fruit was cut 3 cm from its insertion and covered with parafilm. No-Choice Bioassay The infested fruits were sprayed in a Potter’s tower, where each face of the fruit was sprayed with 2 ml of the recommended field concentration for controlling A. guerreronis in coconut palms (MAPA 2017), namely: abamectin (Vertimec 18 EC, 18 g active ingredient (ai)/liter, emulsifiable concentrate, Syngenta, São Paulo, Brazil) at 13.5 mg ai/liter, azadirachtin (AzaMax, 12 g ai/liter, emulsifiable concentrate, VAD Brazil, SP Campinas, Brazil) at 30 mg ai/liter, and fenpyroximate (Ortus 50 SC, 50 g ai/liter, suspension concentrate, Arysta LifeScience, Salto de Pirapora, SP, Brazil) at 100 mg ai/liter, with the solution always totaling 6 ml per fruit. Control was performed with distilled water. The fruits were subsequently allowed to dry at room temperature for approximately 30 min. After this period, 30 adult females of N. baraki were marked on the opisthosoma with fluorescent paint (Blacklight Reactive Fluorescent Acrylic Paint), applied with a No. 000 brush with a single hair. These females were released on the spikelet of each fruit. Five replicates (i.e., fruits) were used per treatment. After 4 h of the mite release, the number of mites marked with fluorescent paint present on the surface of the fruit was quantified. The bracts were then removed with a pruning shear and a staple remover, and the mites marked with fluorescent paint under the bracts were counted. A 20 W ultraviolet lamp (Blacklight, Ourolux, 50/60 Hz) was used to identify the marked mites. Free-Choice Bioassay The experimental unit was similar to the experiment with no chance of choice; however, eight infested fruits were used per treatment, four of which were treated with acaricide and four with distilled water. The spraying of fruits and marking of mites was similar to the no-choice bioassay. The fruits (treated fruit with distilled water + acaricide treated fruit) were joined by the ends of the spikelets with the aid of an entomological pin and parafilm. The distance between the fruits was 10 cm. Thirty adult females of N. baraki were released at the meeting point between the spikelets. Four replicates (i.e., water-treated fruit + acaricide-treated fruit) were used per treatment. After 24 h, the evaluation was performed, as in the no-choice bioassay, for the treated and non-treated fruits. Release of N. baraki on the Bunch The experimental units in this case were clusters of coconut fruits aged 3 to 4 mo (leaf 13 to 14), according to the coconut phyllotaxis (Sobral 1994), whose fruits had 16 to 32% damage intensity (necrosis) by A. guerreronis (Galvão et al. 2008). These bunches were collected on the Island of Itamaracá, PE (07°46′S, 34°52′W), transported to the Laboratory of Acarology of UFRPE and used on the same day to install the experiment. The bunch was cut at 30 cm from the basal part of the bunch rachis before the first spikelet insertion, and 10 fruits infested with the coconut mite A. guerreronis were left per bunch (one per spikelet). A plastic bucket with a capacity of 10 liters was filled with gypsum diluted in water to form a support base for the bunch. Then, the bunch rachis was inserted into a central hole made in the plaster base so that the bunch remained erect. A barrier with entomological glue was made around the base of the rachis. Each bunch was sprayed with the aid of a backpack sprayer, with 300 ml of the recommended field dose for the control of A. guerreronis in coconut palms of the acaricides using either abamectin (13.5 mg ai/liter), azadirachtin (30 mg ai/liter), or fenpyroximate (100 mg ai/liter); a control treatment was sprayed only with distilled water. The sprayed bunch was allowed to dry at room temperature for 45 min. Finally, 100 adult N. baraki females marked with fluorescent ink were released above the glue barrier, as per the previous experiment. After 24 h, the fruits were removed, and the number of mites marked with fluorescent paint was quantified as described for the previous bioassays. Four replicates (i.e., bunches) were used per treatment. Statistical Analyses The mean number of fluorescence-tagged mites recaptured from each treatment in the different release bioassays (fruits spikelet and bunch rachis) were submitted to normality (Kolmogorov’s test) and homogeneity tests (Barlett’s test). The mean mite counts (at the fruit surface and under the bracts) were compared by Student’s t-test (Satterthwaite Method: Equality of Variance) using Proc TTEST (SAS Institute 2008). To determine the difference between the treatments and the control in the mean of recaptured mites in the two regions (under the bracts and surface of the fruit), in the bioassays of release in the spikelet and bunch rachis, the data were submitted to analysis of variance and the Tukey’s HSD test (P < 0.05) (proc ANOVA; SAS Institute 2008). For the free-choice bioassay, the difference in the mean number of marked mites found in the treated and non-treated fruits was compared by the χ2 test (P < 0.05) using the procedure FREQ from SAS (SAS Institute 2008). Results Release and Recapture of N. baraki Free-Choice Bioassays on Fruits Mite recapture was in the range between 40 and 60% with the other initially released mites leaving the coconut arena. Regardless, a significant difference between the number of mites recaptured on the fruit surface and under the bracts was observed in all treatments: control (t5.3 = 4.54, P = 0.005), azadirachtin (t7.9 = 6.34, P = 0.0002), abamectin (t7.72 = −9.07, P < 0.0001), and fenpyroximate (t7.06 = −4.64, P = 0.002). A statistically significant difference was also observed among acaricides for the number of recaptured mites under the bracts (F3.16 = 5.75, P = 0.007). Fenpyroximate differed from the other treatments, exhibiting a lower number of recaptured mites under the bracts. However, no significant difference was observed among the treatments for the number of recaptured mites on the fruit surface (F3.16 = 2.23, P = 0.12) (Fig. 1). Fig. 1. Open in new tabDownload slide Number (±SE) of Neoseiulus baraki recaptured on the surface and under the bracts of the fruits (perianth) after release in the spikelet (evaluation after 4 h). Bars with different capital letters indicate significant differences between the two fruit parts (surface and bract) within the treatment; different lowercase letters indicate significant differences among acaricidal treatments within a given the fruit part (P < 0.05). Fig. 1. Open in new tabDownload slide Number (±SE) of Neoseiulus baraki recaptured on the surface and under the bracts of the fruits (perianth) after release in the spikelet (evaluation after 4 h). Bars with different capital letters indicate significant differences between the two fruit parts (surface and bract) within the treatment; different lowercase letters indicate significant differences among acaricidal treatments within a given the fruit part (P < 0.05). No-choice Bioassays on Fruit Again mite recapture was in the range of 40 to 60%, as in the previous (free-choice) bioassay with the other mites leaving the arena. Among the recaptured mites, most of them dispersed to the non-treated fruits (control). Thus, a statistically significant difference existed between the number of mites recaptured in the non-treated fruit (control) and the abamectin-treated fruit (χ2 = 39.09, df = 1, P < 0.001), between the control and the azadirachtin-treated fruit (χ2 = 32.40, df = 1, P < 0.001), and between the control and the fenpyroximate-treated fruit (χ2 = 28.90, df = 1, P < 0.001) (Fig. 2). Fig. 2. Open in new tabDownload slide Number of predatory mites (Neoseiulus baraki) recaptured in free-choice bioassays between non-treated and acaricide-treated fruits. Asterisks indicate significant difference between non-treated and acaricide-treated fruits (P < 0.05). Fig. 2. Open in new tabDownload slide Number of predatory mites (Neoseiulus baraki) recaptured in free-choice bioassays between non-treated and acaricide-treated fruits. Asterisks indicate significant difference between non-treated and acaricide-treated fruits (P < 0.05). Bioassays on the Bunch Mite recapture was smaller on the bioassay with coconut bunches, between 5 and 40%, as expected due to the greater chance of escaping unrecorded from the fruits of the bunch. Nonetheless, the recapture achieved was large enough to allow recognition of differences among treatments. A significant difference was observed in all treatments between the mean number of mites recaptured on the fruit surface and under the bracts: control (t3.09 = 10.17, P = 0.002), azadirachtin (t4.64 = 7.47; P = 0.001), abamectin (t3.51 = 6.18; P = 0.005), and fenpyroximate (t3 = 6.33; P = 0.008) (Fig. 3). A difference also existed between the acaricides in the number of recaptured mites under the bracts (F3,12 = 59.62; P < 0.0001), where all acaricides differed from the control (Fig. 3). The number of recaptured mites in fruits with acaricides was lower than the number of mites recaptured in the control fruits. A higher number of mites were observed under the bracts on the azadirachtin-treated fruits than in the other acaricide-treated fruits. However, no significant difference existed between the acaricide treatments and the control for the number of mites recaptured on the fruit surface (F3,12 = 1.04, P = 0.41) (Fig. 3). Fig. 3. Open in new tabDownload slide Number (±SE) of marked predatory mites (Neoseiulus baraki) recaptured on the surface and under the bracts of the fruits (perianth) after release on the bunch rachis (evaluation after 24 h). Bars with different capital letters indicate significant differences between the two fruit parts (surface and bract) within the treatment; different lowercase letters indicate significant differences among acaricidal treatments within a given the fruit part (P < 0.05). Fig. 3. Open in new tabDownload slide Number (±SE) of marked predatory mites (Neoseiulus baraki) recaptured on the surface and under the bracts of the fruits (perianth) after release on the bunch rachis (evaluation after 24 h). Bars with different capital letters indicate significant differences between the two fruit parts (surface and bract) within the treatment; different lowercase letters indicate significant differences among acaricidal treatments within a given the fruit part (P < 0.05). Discussion The results show that 1) N. baraki dispersed to fruits not sprayed with acaricides; 2) among the acaricides tested, azadirachtin was the one least-affecting fruit colonization by N. baraki; and 3) the bracts show a higher probability of colonization by the predator, regardless of the presence of acaricide residues. The free-choice bioassay demonstrated that the predatory mites exhibit preferential dispersal to the non-treated surface. According to Lima et al. (2013a), the irritability caused by acaricides may favor the escape of N. baraki to avoid contaminated surfaces. These authors determined that azadirachtin and fenpyroximate cause irritability to N. baraki when exposed to the treated surface, unlike abamectin. However, abamectin affected the speed and distance traveled by N. baraki, with a reduction in mobility. In the bioassay with N. baraki released on the spikelet of fruits under no-choice conditions, no significant difference was observed between the number of recaptured mites under the bracts of non-treated fruits and fruits contaminated with abamectin and azadirachtin. Fenpyroximate differed from the other acaricides, leading to a low recapture of N. baraki under the bracts. According to Lima et al. (2013a,b), fenpyroximate, which is an acaricide selective for this predator, does not affect the walking of N. baraki. Therefeore, N. baraki should have shown greater mobility resulting in dispersal from the experimental system and, consequently, low colonization of the fruit explaining the low recapture achieved with this acaricide in the present study. In the release experiments on the bunch rachis, a larger quantity of N. baraki was recaptured in the region under the bracts. This predatory mite is a generalist and lives in confined spaces of monocotyledonous plants (Subtype III-d) (McMurtry et al. 2013). N. baraki, in order to escape the surface contaminated with acaricide left the experimental system or sought its habitat, which is the region of the fruit protected by the bracts. The bracts of the fruit confer protection against acaricides for the species that inhabit the perianth, such as A. guerreronis (Silva et al. 2017), and also N. baraki. The number of mites recaptured in the acaricide treatments differed statistically from the control, which may be due to the change in the foraging capacity of N. baraki caused by acaricides, which leads to the inability of the predator to distinguish fruits that are infested from those not infested with A. guerreronis (Lima et al. 2013a, 2016). Interestingly, azadirachtin stood out from the other acaricides because of allowing a higher number of recaptured mites. Despite the irritability that azadirachtin causes (Lima et al. 2013a), N. baraki was able to access the bracts instead of leaving the experimental system. Most likely, this occurred because of the ability of N. baraki to escape and avoid surfaces contaminated with azadirachtin, as reported by Lima et al. (2013a), accessing the region under the bracts free of acaricide residues. Possibly, the process of walking dispersal within the same bunch involves relatively small distances for this predator, allowing it to penetrate the perianth of other fruits of the same bunch before even trying to passively leave the bunch via wind transport. The effects caused by sublethal acaricide concentrations on N. baraki may compromise its effectiveness for biological control (Lima et al. 2013a, 2016). Insecticide exposure causes N. baraki to avoid contaminated areas and also favors the predator aerial dispersal, effect particularly strong with azadirachtin (Lima et al. 2013a, Monteiro et al. 2018). On the other hand, once the predators reach the perianth region (under the bracts), they are protected from acaricide exposure (Silva et al. 2017). Thus, sublethal concentrations may cause N. baraki to remain under the bracts for longer and, consequently, may result in reduction of the local population of A. guerreronis. However, some acaricides like abamectin and fenpyroximate compromise the population growth of N. baraki, and these effects are extended to the first generation (Lima et al. 2016). This emphasizes the need of comprehensive assessments to recognize the impact and compatibility of these compounds with this biocontrol agent. The presence of acaricides negatively affected the colonization by N. baraki, what may affect the efficiency of the natural biological control of the coconut mite A. guerreronis in the field (Lima et al. 2013a,b; Monteiro 2018; and present study). 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This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) TI - Acaricide-Mediated Colonization of Mite-Infested Coconuts by the Predatory Phytoseiid Neoseiulus baraki (Acari: Phytoseiidae) JF - Journal of Economic Entomology DO - 10.1093/jee/toy291 DA - 2019-02-12 UR - https://www.deepdyve.com/lp/oxford-university-press/acaricide-mediated-colonization-of-mite-infested-coconuts-by-the-m8JaB9HrqD SP - 213 VL - 112 IS - 1 DP - DeepDyve ER -