TY - JOUR
AU - Pavan,, Francesco
AB - Abstract The effects of kaolin and bunch-zone leaf removal on populations of predatory mites Kampimodromus aberrans (Oudemans) and Typhlodromus pyri Scheuten were assessed in the context of four trials (2015–2016) carried out against Lobesia botrana (Denis and Schiffermüller) (Lepidoptera: Tortricidae) in vineyards located in north-eastern Italy. Laboratory experiments were performed to evaluate the effects of kaolin on the survival and fecundity of K. aberrans and T. pyri populations originating from the same grape-growing area. In field trials, kaolin caused a gradual decrease in population density levels of both phytoseiid species (with the maximum reduction ranging from 49 to 91%) with a complete population recovery in the next spring. In laboratory experiments, kaolin was moderately harmful to both species, reducing their fecundity significantly (around 60%). Bunch-zone leaf removal determined lower phytoseiid populations in all trials, but this effect was significant only for K. aberrans in one of them. A limited use of kaolin and the adoption of bunch-zone leaf removal did not irreversibly affect phytoseiid populations in vineyards and thus can be considered compatible with IPM strategies. Kampimodromus aberrans, Typhlodromus pyri, natural enemy, grapevine, cultural practice Kaolin is at the basis of particle film technology (Glenn et al. 1999), which consists of covering plants with a thin layer of this product. It is a cultural practice used to enhance tolerance to water stress and improve water productivity (Boari et al. 2015). It also acts as a physical barrier for arthropods, hiding the host plant visually or chemically, and thus, deterring egg laying and feeding, with negative effects on reproduction, survival, and development (Unruh et al. 2000, Vincent et al. 2003, Barker et al. 2006, Lapointe et al. 2006, Tacoli et al. 2017b). This technology has been reported as effective in reducing the infestation of several insect and mite pests (e.g., Knight et al. 2001, Glenn and Puterka 2005, Jaastad et al. 2006, Markó et al. 2008, Pascual et al. 2010, D’Aquino et al. 2011, Tyler-Julian et al. 2014, Marcotegui et al. 2015). In vineyards, kaolin is sprayed to optimize plant water use efficiency and reduce berry sunburn (Brillante et al. 2016, Ferrari et al. 2017). However, it is also effective in the control of hemipteran pests, such as the sharpshooter Homalodisca coagulata (Say), which is vector of Xylella fastidiosa Wells, the causal agent of Pierce’s disease (Wood and McBride 2001, Puterka et al. 2003, Barker et al. 2006, Tubajika et al. 2007), the leafhoppers Empoasca vitis (Göthe), Zygina rhamni Ferrari and Scaphoideus titanus Ball (Tacoli et al. 2017a,b), the grape phylloxera Daktulosphaira vitifoliae (Fitch) (Sleezer et al. 2011), and the vine cicada Psalmocharias alhageos (Kolenati) (Valizadeh et al. 2013). Kaolin is also active against the tortricid moth Lobesia botrana (Denis and Schiffermüller) (Lepidoptera: Tortricidae) (Pease et al. 2016, Tacoli et al. 2018). Even bunch-zone leaf removal, consisting in removing all the leaves that cover bunches, can reduce L. botrana infestation (Pavan et al. 2016) and a positive synergy with kaolin in the control of this tortricid pest was reported (Tacoli et al. 2018). This cultural practice can favor sunburn of berries exposed to sunlight (Mosetti et al. 2016) and kaolin can compensate for this disadvantage. Kaolin is proposed as a viable alternative to synthetic insecticides in vineyards. However, the preservation of biological control agents is essential in the framework of integrated pest management (IPM) and knowledge is limited about kaolin side effects on beneficials. Kaolin did not negatively affect the parasitization of eggs of the grapevine leafhoppers E. vitis and Z. rhamni by Anagrus spp. (Hymenoptera: Mymaridae) in vineyards, nor did it reduce parasitization of L. botrana eggs by Trichogramma cacoeciae Marchal (Hymenoptera: Trichogrammatidae) in the laboratory (Pease et al. 2016, Tacoli et al. 2017b). Even bunch-zone leaf removal, altering the grapevine canopy structure and microclimate (Kiaeian Moosavi et al. 2018), could affect nontarget arthropods. In vineyards, predatory mites belonging to the family Phytoseiidae effectively control tetranychid and eriophyoid mites (e.g., Pérez-Moreno and Moraza-Zorrilla 1998, Duso et al. 2012, McMurtry et al. 2013, Tixier et al. 2013), but their activity can be affected by fungicides and broad-spectrum insecticides (e.g., Duso 1994; Pozzebon et al. 2002, 2010, 2015; Tirello et al. 2013). For this reason, phytoseiid mites are considered as representative environmental indicators of the impact of pesticides in agriculture and the selectivity of pesticides to predatory mites (and other beneficials) is carefully evaluated by regulatory organizations (for Europe see, Candolfi et al. 1999). Negative side effects of kaolin on phytoseiid mites have been recorded in American and European orchards (Benedict 2005, Jaastad et al. 2006, Villanueva and Walgenbach 2010), although Bostanian and Racette (2008) did not observe such effects under laboratory conditions. No influence of canopy architecture on the density of generalist phytoseiids was previously found (Prishmann et al. 2006). In the current study, the side effects of kaolin were evaluated on Kampimodromus aberrans (Oudemans) and Typhlodromus pyri Scheuten populations under field and laboratory conditions. These species are key biocontrol agents of phytophagous mites in vineyards and orchards and nontarget species in the evaluation of pesticide side effects (Candolfi et al. 1999; Duso et al. 2009; Pozzebon et al. 2010, 2015; Tirello et al. 2013; Wearing 2014; Ahmad et al. 2015). The impact of the bunch-zone leaf removal on predatory mite abundance was also evaluated. Materials and Methods Field Trials The influence of kaolin and bunch-zone leaf removal on predatory mites was studied in the framework of four field trials against L. botrana (Tacoli et al. 2018) carried out in three vineyards in north-eastern Italy (Gorizia district, cultivar Pinot Gris), designated as Vineyard A (2015 and 2016 growing seasons), Vineyard B (2015), and Vineyard C (2016). Vineyard A (45°57′51″N, 13°26′49″E, 56 m a.s.l.) is a 10-yr-old conventional vineyard with vines grown using the Guyot system with distances between and along rows of 2.5 and 0.8 m, respectively. Vineyard B (45°57′20″N, 13°26′50″E, 50 m a.s.l.) is a 30-yr-old organic vineyard with vines grown using the double arched Guyot system with distances between and along rows of 2.8 and 1 m, respectively. Vineyard C (45°58′02″N, 13°31′31″E, 53 m a.s.l.) is a 15-yr-old organic vineyard with vines grown using the Guyot system with distances between and along rows of 2.2 and 0.7 m, respectively. In experimental plots, standard fungicide programs were followed and insecticides were not applied. In the four trials (Vineyard A 2015, Vineyard B 2015, Vineyard A 2016, and Vineyard C 2016), kaolin (Surround WP, Tessenderlo Kerley Inc., Phoenix, AZ, 2% w/v, Surround WP/water) was sprayed at the rate of 1,000 liter/ha, and an untreated control was included for a comparison. Kaolin was applied twice in 2015 (18 and 24 June both in Vineyards A and B) and three times in 2016 (10, 20, and 28 June in Vineyard A; 10 and 24 June and 1 July in Vineyard C). Although two applications of kaolin were sufficient to ensure satisfactory bunch coverage and consequently good L. botrana control, in 2016, a further application was carried out as the product had been partially washed off by rain (Tacoli et al. 2018). In all cases, kaolin was applied with a backpack sprayer (Oleo-Mac, Sp-126, Emak S.p.A., Bagnolo in Piano, RE, Italy). In all trials, a randomized block design with four replicates was adopted. Each block consisted of a vineyard row divided into two plots (kaolin and control) of 28 (Vineyard A both 2015 and 2016), 20 (Vineyard B), or 24 (Vineyard C) vines. Plots were divided into two subplots of 14 (Vineyard A), 10 (Vineyard B), or 12 (Vineyard C) vines that were subjected or not to bunch-zone leaf removal (17 June 2015 and 10 June 2016). In Vineyard A, the plots and subplots submitted, respectively, to kaolin applications and bunch-zone leaf removal were the same in both years. Preliminary observations revealed that the dominant species were K. aberrans in Vineyards A and B and T. pyri in Vineyard C. To assess phytoseiid densities, five samplings were carried out in each year and trial (2015: 11, 22, and 29 June, 06 July, and 20 August both in Vineyard A and Vineyard B; 2016: 6, 20, and 28 June, 6 July, and 24 August in Vineyard A; 7 and 21 June, 1 and 8 July, and 24 August in Vineyard C). On each sampling date, 10 leaves were collected from the mid parts of the main vine shoots in each subplot (40 leaves per subplot); they were enclosed in plastic bags and cool stored until being transferred to the laboratory. The leaves were checked under a dissecting microscope to assess mite numbers. At least 100 specimens per trial were slide mounted in Berlese medium and identified under 400× magnification, using current keys (Cargnus et al. 2012, Tixier et al. 2013). In Vineyard A, overwintering phytoseiid populations were sampled (5 December 2016) after two successive years (2015 and 2016) of kaolin applications on the same vines. Pruned 2-yr-old cane portions consisting of three nodes and internodes were collected from 10 grapevines per replicate (subplot without bunch-zone leaf removal) for a total of 40 cane portions per treatment. The number of predatory mites, overwintering as adult females under the bark and inside the buds, was counted under a dissecting microscope. At least 100 specimens were prepared as above to allow the identification at species level (Cargnus et al. 2012, Tixier et al. 2013). Laboratory Experiments Laboratory experiments were performed to evaluate the effects of kaolin on K. aberrans and T. pyri populations originating, respectively, from Vineyard A and an organic vineyard (Udine district, 46°06′45″N, 13°24′40″E, 140 m a.s.l., cultivar Merlot) located in the same grape-growing area as Vineyard C. Predatory mites were reared in the laboratory for some generations according to Tirello et al. (2013). For both species, toxicological tests were performed using insecticide-free grapevine leaf discs (4.5 cm in diameter). Kaolin (Surround WP, 4% w/v, Surround WP/water) was applied to leaf discs with a Potter spray tower (Burkard Scientific Ltd, Uxbridge, United Kingdom) spraying 1.4 ml of suspension per leaf disc at 103 kPa (15 psi) to obtain an amount of fluid of 1.9–2.0 mg/cm2, as recommended by the IOBC guidelines (Sterk et al. 1999). The untreated leaf discs (control) were sprayed with water following the same procedure. Leaf discs were subsequently placed on wet cotton pads and wet cotton barriers were created along their perimeter to prevent predatory mites from escaping. Two mated females (about 12-d old) were placed on each leaf disc. Fresh pollen was provided every 2 d as food. The experiments were conducted under controlled conditions (25°C, 70% RH, and photoperiod of 16:8 [L:D] h). Toxicological tests were run following Tirello et al. (2013). Female mortality was checked at 72 h after spraying and fecundity was assessed daily for four additional days. After 7 d, the remaining females and juvenile stages were removed and eggs were monitored until they had completely hatched in the control. In total, 50 and 38 females per treatment were assessed for K. aberrans and T. pyri, respectively. Statistical Analyses To compare field data, one-way and mixed analysis of variance (ANOVA) with Bonferroni adjustment and Tukey’s post hoc test were performed after logarithmic transformation using IBM SPSS Statistics 20 (SPSS 2011). The reduction effect of kaolin was calculated according to Henderson and Tilton (1955). To compare field data in the sampling before the first kaolin application, a Student’s unpaired t-test was used. Data on fecundity were analyzed with a one-way ANOVA using GLM procedure of SAS (SAS Institute Inc. 1999) considering leaf disks treatment as source of variation and testing its effect with an F-test (α = 0.05). A Tukey’s test (α = 0.001) was used in post hoc analysis to test differences among means. The data on fecundity were square-root transformed prior to the analyses, in order to meet the ANOVA assumptions. Possible changes in the number of females present on the test units during the reproduction period and the hatching of larvae from eggs between the assessment dates were taken into account by using the Blümel and Hausdorf (2002) formula with some modifications, considering the number of eggs laid from the first day: Rry= [nEd3(nFd3+ nFd1)/2]+ ∑7 x = 4 [nEdx− nEdx−1(nFdx+ nFdx−1)/2] where Rry is the reproduction in replicate number y; d1, d3, and dx are evaluation days; nEdx is the number of eggs (in replicate number y) on day x; and nFdx is the number of females (in replicate number y) on day x. The overall effect of kaolin was expressed as E = 100% − (100% − M) × R (Tirello et al. 2013), where E is the coefficient of toxicity; M is the mortality percentage of females calculated following Abbott (1925); and R is the ratio between the average number of hatched eggs produced by females in kaolin treatment and the average number of hatched eggs produced by females in the control treatment. Results Field Trials Vineyard A 2015 and 2016 In both years, K. aberrans was the only species recorded and its population density in the control plots ranged from 6.0 to 17.0 motile forms per leaf. In both years, phytophagous mite densities were negligible. In both years, K. aberrans densities were not significantly different in the two treatments in the sampling made before the first kaolin application (2015: t = 0.69; df = 6; P = 0.52; 2016: t = 1.49; df = 6; P = 0.19; Fig. 1). After kaolin applications, phytoseiid densities were significantly lower in the kaolin than in the control plots (2015: F = 9.013; df = 1,12; P = 0.011; 2016: F = 80.802; df = 1,12; P = 0.0001; Fig. 1). Kampimodromus aberrans densities became significantly lower in the kaolin compared with the control plots after the second application in 2015 and after the first application in 2016 (i.e., 18 and 11 d after first application, respectively). The reduction effect of kaolin at about 10 d from the last application was higher in 2016 (Henderson-Tilton reduction of 69%) than in 2015 (49%). After two successive years of kaolin applications (2015 and 2016) in the same plots, the overwintered populations were not significantly different between the kaolin and the control (F = 0.735; df = 1; P = 0.424; Fig. 2). Fig. 1. Open in new tabDownload slide Number of phytoseiid mites recorded in kaolin and the control plots of the four trials before the first (early June) and after two (2015) or three (2016) kaolin applications (arrows). NS, *, **, *** indicate nonsignificant and significant differences for α = 0.05, α = 0.01¸ α = 0.0001, respectively, between treatments according to Tukey’s post hoc test. Fig. 1. Open in new tabDownload slide Number of phytoseiid mites recorded in kaolin and the control plots of the four trials before the first (early June) and after two (2015) or three (2016) kaolin applications (arrows). NS, *, **, *** indicate nonsignificant and significant differences for α = 0.05, α = 0.01¸ α = 0.0001, respectively, between treatments according to Tukey’s post hoc test. Fig. 2. Open in new tabDownload slide Number of overwintered females of the phytoseiid mite K. aberrans (mean ± SD) recorded in the kaolin and the control plots in December 2016 in Vineyard A. NS indicates nonsignificant differences between treatments according to one-way ANOVA. Fig. 2. Open in new tabDownload slide Number of overwintered females of the phytoseiid mite K. aberrans (mean ± SD) recorded in the kaolin and the control plots in December 2016 in Vineyard A. NS indicates nonsignificant differences between treatments according to one-way ANOVA. In both years, bunch-zone leaf removal did not influence significantly phytoseiid densities (2015: F = 1.029; df = 1,12; P = 0.33; 2016: F = 4.573; df = 1,12; P = 0.054; Fig. 3). The effect of the interaction ‘kaolin * bunch-zone leaf removal’ was not significant (2015: F = 1.120; df = 1,12; P = 0.311; 2016: F = 0.35; df = 1,12; P = 0.855). Fig. 3. Open in new tabDownload slide Number of phytoseiid mites recorded in the four trials in the subplots with and without bunch-zone leaf removal, which was carried out the day after the first sampling date. NS and ** indicate, respectively, nonsignificant and significant differences for α = 0.01 between treatments according to Tukey’s post hoc test. Fig. 3. Open in new tabDownload slide Number of phytoseiid mites recorded in the four trials in the subplots with and without bunch-zone leaf removal, which was carried out the day after the first sampling date. NS and ** indicate, respectively, nonsignificant and significant differences for α = 0.01 between treatments according to Tukey’s post hoc test. Vineyard B Phytoseiid identification revealed the dominance of K. aberrans (89%) over Amblyseius andersoni (Chant) (11%) and their population densities in the control plots ranged from 1.8 to 2.3 motile forms per leaf. Phytophagous mite densities were negligible. In the sampling before the first kaolin application, phytoseiid densities in kaolin and the control plots were not significantly different (t = 0.40; df = 6; P = 0.70; Fig. 1). After two kaolin applications, phytoseiid densities were significantly lower in kaolin than in the control plots (F = 16.575; df = 1,12; P = 0.002; Fig. 1). In particular, K. aberrans densities were significantly lower in kaolin plots 10 d after the first application. The reduction effect of kaolin at about 10 d from the last application was substantial (Henderson–Tilton reduction: 91%). Bunch-zone leaf removal significantly reduced phytoseiid densities (F = 11.926; df = 1,12; P = 0.005; Fig. 3). In particular, the negative effect of bunch-zone leaf removal was significant in July and August. However, the negative effect of this practice (maximum Henderson–Tilton reduction of 34% in the last sampling) was lower than the kaolin applications. The effect of the interaction ‘kaolin * bunch-zone leaf removal’ was not significant (F = 0.499; df = 1,12; P = 0.493). Vineyard C Typhlodromus pyri (97%) dominated over A. andersoni (3%) and their population densities in the control plots ranged from 0.6 to 1.6 motile forms per leaf. Phytophagous mite densities were negligible. In the sampling before the first kaolin application, T. pyri densities in kaolin and the control plots were not significantly different (t = 0.70; df = 6; P = 0.51; Fig. 1). Kaolin applications significantly reduced phytoseiid densities in the kaolin plots compared with the control plots (F = 8.218; df = 1,12; P = 0.014; Fig. 1). Typhlodromus pyri densities were significantly lower in the kaolin plots 7 d after the second kaolin application (i.e., 24 d after first application). As in Vineyard B, the negative effect of kaolin at about 10 d from the last application was substantial (Henderson–Tilton reduction of 88%). Bunch-zone leaf removal did not significantly reduce phytoseiid densities (F = 1.577; df = 1,12; P = 0.233; Fig. 3) and the interaction ‘kaolin * bunch-zone leaf removal’ was not significant (F = 0.044; df = 1,12; P = 0.837). Laboratory Experiments In the control, the fecundity rates (mean ± SD) of both K. aberrans and T. pyri were 0.78 ± 0.23 and 0.88 ± 0.14 eggs/female/day, respectively. Kampimodromus aberrans female survival was 100% in both treatments but kaolin significantly reduced fecundity (F = 124.78; df = 45; P < 0.0001; Fig. 4) affecting the toxicity coefficient E (62.8%). In both treatments, the hatching rate was 100%. Fig. 4. Open in new tabDownload slide Daily number of eggs (mean ± SD) laid on grapevine leaf discs by females of the two phytoseiid species in the kaolin and the control. Different letters indicate significant differences for α = 0.001 according to Tukey’s post hoc test. Fig. 4. Open in new tabDownload slide Daily number of eggs (mean ± SD) laid on grapevine leaf discs by females of the two phytoseiid species in the kaolin and the control. Different letters indicate significant differences for α = 0.001 according to Tukey’s post hoc test. Typhlodromus pyri female survival was 100% in both treatments. However, kaolin significantly reduced female fecundity (F = 94.26; df = 33; P < 0.0001; Fig. 4) affecting the toxicity coefficient E (62.5%). In both treatments, the hatching rate was 100%. Discussion In field trials, kaolin affected K. aberrans and T. pyri populations negatively. For both species, the differences between treatments reached maximum levels on the second or third sampling date after the first kaolin application, i.e., after about 20 d. These data suggest that kaolin did not cause knock down effects on predatory mites. The negative effect of kaolin on K. aberrans in Vineyard A did not persist over the two seasons. In fact, the negative effect on K. aberrans densities observed in the summer of 2015 was no longer recorded in the pretrial sampling in 2016 and that observed in the summer of 2016 had no consequence on the abundance of overwintering females. We can suggest that in the kaolin-treated plots the predatory mites moved to kaolin-free leaves, developed after the product applications, where they had more chance to reproduce and multiply. A lower intraspecific competition can be also hypothesized during the late season in kaolin-treated plots (Duso 1989, Bengochea et al. 2013, Ahmad et al. 2015, Pozzebon et al. 2015). The negative effects of kaolin on phytoseiid mites, recorded in the present study, have been reported previously. Trials carried out in two subsequent years in plum and apple orchards in Norway showed that kaolin sprayed against phytophagous mites and tortricid moths affected phytoseiid mites in three out of four cases (Jaastad et al. 2006), even though part of the effect might have been associated with lower prey populations. In an apple orchard located in New England (United States), Benedict (2005) found both significantly higher levels of tetranychid mites and lower levels of phytoseiid mites on trees treated with kaolin compared with those left unsprayed. In Vineyard A, a greater detrimental effect of kaolin on K. aberrans was observed in the trial of 2016 than in that of 2015, probably due to the additional kaolin application done in 2016 (i.e., three instead of two). In 2015 in Vineyard B, the peak of the K. aberrans population was almost 10-fold lower than in Vineyard A. One factor involved in this difference could have been lower pollen availability in Vineyard B, due to less dense inter-row herbaceous vegetation following tillage carried out in early spring. In Vineyard C, T. pyri population densities were lower than those of K. aberrans recorded in Vineyards A and B. This is in agreement with studies that reported a lower density of T. pyri than K. aberrans in similar vineyard conditions (Duso 1989, 1992). T. pyri populations declined in summer probably due to the high susceptibility of this species to high temperatures combined with low relative humidity (Duso et al. 1991, Duso and Pasqualetto 1993). In these conditions, pollen availability declines, and thus predatory mites suffer from food shortage (Duso et al. 1997, Pozzebon et al. 2005). Moreover, the leaf morphology of Pinot Gris leaves (Vineyard C) is a further factor that can explain the low population size of T. pyri in this vineyard (Duso and Vettorazzo 1999, Roda et al. 2003). The impact of kaolin on these populations can represent an additional factor affecting predatory mite persistence and activity. In the laboratory experiments, the fecundity rates of both K. aberrans and T. pyri in the control were comparable with those reported for these species by Lorenzon et al. (2012). Laboratory data showed that kaolin reduced the fecundity of K. aberrans and T. pyri females but not their survival. Therefore, the decrease observed in fecundity did not occur as a consequence of a decrease in survival. The negative effect of kaolin on oviposition could explain the progressive decline in phytoseiid mite densities observed in the field for both species. In contrast with our data, Benedict (2005) reported greater T. pyri mortality on kaolin treated leaves (25%). This effect was associated with a reduction in Tetranychus urticae Koch (Acari: Tetranychidae) availability caused by kaolin, whereas in our study, the mortality did not increase because phytoseiid mites were fed on pollen, similar to the situation in the trial vineyard. In the case of another phytoseiid species, i.e., Neoseiulus fallacis (Garman), Bostanian and Racette (2008) observed no negative effects on either on survival or on fecundity, whereas Benedict (2005) reported a decrease in survival (12%) that was always associated with a lower T. urticae consumption. Bunch-zone leaf removal caused in one out of three cases a significant reduction in K. aberrans population. Reducing canopy density could determine a microclimate characterized by higher temperature and lower relative humidity, known to be less favourable to several phytoseiid mites (Sabelis 1981). However, for T pyri no negative influence of canopy density was observed in agreement with Prishmann et al. (2006). In conclusion, the repeated use of kaolin in viticulture was harmful to phytoseiid populations when these effects were measured within the growing season. Nevertheless, K. aberrans populations recovered after two successive years of kaolin applications. Localizing kaolin applications to the bunch zone when controlling L. botrana could mitigate the impact of kaolin. Based on these outcomes, a moderate use of kaolin and the adoption of leaf removal do not irreversibly affect phytoseiid populations in vineyards and thus can be considered compatible with IPM strategies. Acknowledgments We would like to thank all the vineyard owners who kindly offered their properties as trial sites: Renzo Sgubin, Federico Bigot, and Denis and Patrick Sturm. We would also like to thank all the people who collaborated in the trials: Giovanni Bigot, Davide Cisilino, Davide Mosetti, and Michele Stecchina. References Cited Abbott W. S . 1925 . A method for computing the effectiveness of an insecticide . J. 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TI - Side Effects of Kaolin and Bunch-Zone Leaf Removal on Predatory Mite Populations (Acari: Phytoseiidae) Occurring in Vineyards
JF - Journal of Economic Entomology
DO - 10.1093/jee/toy431
DA - 2019-05-22
UR - https://www.deepdyve.com/lp/oxford-university-press/side-effects-of-kaolin-and-bunch-zone-leaf-removal-on-predatory-mite-0rDGCtKbWf
SP - 1292
VL - 112
IS - 3
DP - DeepDyve
ER -