Temperature Affects Biological Control Efficacy: A Microcosm Study of Trichogramma achaeae.

Current quality control of mass-reared biological control agents (BCAs) is usually performed in the laboratory and often fails to include behavioural aspects of the BCAs. As a result, the use of efficacy measurements determined solely under laboratory conditions to predict field efficacy can be questioned. In this study, microcosms were designed to estimate biological control efficacy (realised parasitisation efficiency) of Trichogramma achaeae Nagaraja and Nagarkatti (Hymenoptera: Trichogrammatidae) parasitising Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs across the operational temperature range (15-30 °C). Temperature greatly affected the success of females in finding and parasitising E. kuehniella eggs, with parasitisation being reduced at 15 and 20 °C, as both the percentage of parasitised host eggs and the percentage of leaves per plant with parasitised host eggs decreased sharply compared with higher temperatures. Graphing previous data on laboratory fecundity against parasitisation efficiency shows that the laboratory-measured fecundity of T. achaeae was unlikely to predict field efficacy across temperatures. Results also showed that leaf side had no effect on the preference of T. achaeae in parasitising E. kuehniella eggs; however, T. achaeae preferred to lay their eggs on the top tier of plants. These findings suggest that more complex assays, which include behavioural responses, might be developed for optimised quality control of BCAs intended for field application.

Influence of the plant interacting entomopathogenic fungus Beauveria bassiana on parasitoid host choice-behavior, development, and plant defense pathways.

Inoculating plants with entomopathogenic fungi may influence plant nutrient uptake and growth, and herbivore performance. Knowledge is limited concerning the effects of this symbiosis on higher trophic levels. We examined how fungal treatment of faba bean seeds with the entomopathogenic fungus Beauveria bassiana influenced the choice-behavior and development of the aphid parasitoid Aphidius colemani. We also sampled plant material for analysis of changes in expression of genes related to plant defense pathways. While parasitoids were compatible with plants inoculated with B. bassiana initially (66 vs. 65% parasitization on inoculated and control plants, respectively; similar development times of parasitoids: 9.2 days), the emergence of adult parasitoids originating from aphids on fungus treated plants was significantly lower (67 vs. 76%, respectively). We also found that the defense response changed, similar to induced systemic resistance, when plants were treated with B. bassiana, similarly to what has been found for other plant symbiotic microorganisms. These novel findings show that although the application of entomopathogenic fungi to plants can alter the plants' defense against herbivores, it may also have an impact on beneficial insects, so their function and use should be evaluated on a case-by-case basis.

Demography and Mass Rearing of Amblyseius swirskii (Acari: Phytoseiidae) Fed on Two Species of Stored-Product Mites and Their Mixture.

Amblyseius swirskii (Athias-Henriot) is one of the most important biological control agents and has been used in many countries. In this study, the suitability of Carpoglyphus lactis L. (Acari: Carpoglyphidae), Tyrophagus putrescentiae (Schrank), and their mixture for rearing A. swirskii was evaluated by using the two-sex life table. When reared on C. lactis, the durations of egg, deutonymph, total preadult, total preoviposition period, and adult male longevity of A. swirskii (1.95, 1.17, 6.13, 6.30, and 10.09 d, respectively) were significantly shorter than those reared on the mixed prey (2.05, 1.45, 6.55, 6.64, and 15.56 d, respectively). When reared on the mixed prey, however, the fecundity (110.21 eggs/female) of A. swirskii was significantly higher in comparison with those solely reared on C. lactis (82.17 eggs/female) or on T. putrescentiae (98.23 eggs/female). When reared on the mixed diet, the intrinsic rate of increase (r = 0.3792 d-1), finite rate of population increase (λ = 1.4611 d-1), and net reproductive rate (R0 = 79.69 offspring) of A. swirskii were significantly higher than those on single-species diets. For a daily production of 10,000 eggs of A. swirskii, a smaller population size (2,626 individuals) of A. swirskii is needed when reared on the mixed diet, in contrast to the 4,332 individuals on C. lactis and 3,778 individuals on T. putrescentiae. The population projection and mass-rearing analysis based on life table showed that the mixed diet was the most suitable and economical diet for the mass-rearing of A. swirskii.

Increased fecundity of Aphis fabae on Vicia faba plants following seed or leaf inoculation with the entomopathogenic fungus Beauveria bassiana.

Since the discovery that entomopathogenic fungi can live inside plants as endophytes, researchers have been trying to understand how this affects mainly plants and herbivores. We studied how inoculation of Vicia faba L. (Fabales: Fabaceae) plants with Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Ascomycota: Hypocreales) (strain GHA) either via the seeds or leaves influenced the nymph production of two successive generations of Aphis fabae Scopoli (Hemiptera: Aphididae). While we did not find any difference in nymph production for the first generation of aphids, second-generation aphids on both seed- and spray inoculated plants produced significantly higher numbers of nymphs than aphids on uninoculated plants. This emphasizes the importance of two (or multi-) generational experimentation. Beauveria bassiana was recovered from 26.0, 68.8 and 6.3% of respectively seed-, spray inoculated and control plants, thus, demonstrating its ability to live as an endophyte in V. faba. The confirmation that plants inoculated with entomopathogenic fungi can have a positive effect on pest insects makes careful consideration of these multi-trophic interactions imperative.

Role of the aphid species and their feeding locations in parasitization behavior of Aphelinus abdominalis, a parasitoid of the lettuce aphid Nasonovia ribisnigri.

Aphid species feeding on lettuce occupy distinct feeding sites: the lettuce aphid Nasonovia ribisnigri prefers to feed on heart leaves, whereas the potato aphid Macrosiphum euphorbiae feeds only on outer leaves. The aphid parasitoid Aphelinus abdominalis, known to be able to regulate M. euphorbiae on many crops, has recently been indicated as a promising biocontrol candidate also for use against N. ribisnigri, a major pest of lettuce. This study therefore examined A. abdominalis parasitization preference between N. ribisnigri and M. euphorbiae and its ability to parasitize aphids feeding on different parts of lettuce plants. In addition, life history traits of A. abdominalis on these aphid species were investigated. In no-choice laboratory experiments on leaf discs and 24 h exposure, A. abdominalis successfully parasitized 54% and 60% of the offered N. ribisnigri and M. euphorbiae, respectively, with no significant difference. In the corresponding choice experiment, however, A. abdominalis had a tendency for a significantly higher preference for M. euphorbiae (38%) compared to N. ribisnigri (30%). Growth chamber experiments on whole plants demonstrated that A. abdominalis was able to parasitize aphids, regardless of their feeding locations on lettuce plants. However, aphid feeding behavior had a significant effect on the parasitization rate. A. abdominalis parasitized significantly higher percentages of M. euphorbiae or N. ribisnigri when aphids were exposed separately to parasitoids on whole lettuce plants as compared with N. ribisnigri exposed only on heart leaf. A significant preference of A. abdominalis for M. euphorbiae compared to N. ribisnigri was also observed in the growth chamber choice experiment. A high percentage of adult emergence (> 84%) and female-biased sex ratio (> 83%) were found irrespective of the aphid species.

Sublethal and Transgenerational Effects of Abamectin on the Biological Performance of the Predatory Thrips Scolothrips longicornis (Thysanoptera: Thripidae).

Determination of sublethal and transgenerational effects of pesticides on natural enemies is necessary for successful implementation of biocontrol in integrated pest management programs. In this study, these effects of abamectin on the predatory thrips Scolothrips longicornis Priesner fed on Tetranychus urticae Koch were estimated under laboratory conditions in which adult predators were exposed to pesticide residues on bean leaves. The estimated values of LC50 for female and male predators were 0.091 and 0.067 µg a.i./ml, respectively. Low-lethal concentrations (LC10, LC20 and LC30) of abamectin severely affected fecundity and longevity of treated females of S. longicornis. In addition, transgenerational effects on reproductive and life table parameters of the subsequent generation were observed. The results from this research can be used to develop guidelines for the use of abamectin to minimize the impact on S. longicornis.

The effects of strawberry cropping practices on the strawberry tortricid (Lepidoptera: Tortricidae), its natural enemies, and the presence of nematodes.

Cropping practice can affect pests and natural enemies. A three-year study of the strawberry tortricid, Acleris comariana (Lienig and Zeller) (Lepidoptera: Tortricidae), its parasitoid Copidosoma aretas Walker (Hymenoptera: Encyrtidae), and its entomopathogenic fungi was conducted in seven pairs of organic and conventional farms to test the hypothesis that farming practice (organic versus conventional) will affect the level of pest infestation and will affect the natural enemies. In addition, the number of years with strawberries on the farm, field age, and other factors that may affect pests and their natural enemies were considered. Farms were characterized by their cropping practices, cropping history, and other parameters. Field-collected larvae were laboratory reared to assess mortality from parasitoids and entomopathogenic fungi. In 2010, a survey of nematodes was made to assess the response of an unrelated taxonomic group to cropping practice. 2,743 larvae were collected. Of those, 2,584 were identified as A. comariana. 579 A. comariana were parasitized by C. aretas and 64 A. comariana were parasitized by other parasitoid species. Finally, 28% of the larvae and pupae of A. comariana died from unknown causes. Only two of the field-collected A. comariana larvae were infected by entomopathogenic fungi; one was infected by Isaria sp. and the other by Beauvaria sp. The density of A. comariana was on average four times lower in organic farms, which was significantly lower than in conventional farms. A. comariana was more dominant on conventional farms than on organic farms. The effect of crop age (One, two, or three years) on A. comariana infestation was significant, with higher infestations in older fields. Crop age had no effect on A. comariana infestation in a comparison of first- and second-year fields in 2010. Cropping practice did not lead to significant differences in the level of total parasitism or in C. aretas parasitism; however, C. aretas contributed to a higher proportion of the parasitized larvae on conventional farms than on organic farms. Mortality from unknown causes of A. comariana was higher in organic farms than conventional farms, and unknown mortality was two to seven times higher in second-generation A. comariana than in first generation. Entomopathogenic nematodes were found on three organic farms and one conventional farm. Plant parasitic nematodes were found in more samples from conventional farms than from organic farms. The low density of A. comariana in organic farms exposes the specialist C. aretas to a higher risk of local extinction. In organic farms, where the density of A. comariana is low, other parasitoids may play an important role in controlling A. comariana by supplementing C. aretas. Other tortricid species may serve as alternative hosts for these other parasitoids, contributing to conserving them in the habitat. The higher unknown mortality of larvae from organic fields may be the result of non-consumptive parasitoid or predator effects. This study reports an example of the effects of cropping practice on an insect pest, with similar effects on nematodes. An understanding of the responsible factors could be used to develop more sustainable cropping systems.

Influence of cornicle droplet secretions of the cabbage aphid, Brevicoryne brassicae, on parasitism behavior of naïve and experienced Diaeretiella rapae.

Insects have evolved amazing methods of defense to ward off enemies. Many aphids release cornicle secretions when attacked by predators and parasitoids. These secretions contain an alarm pheromone that alerts other colony members of danger, thereby providing indirect fitness benefits to the releaser. In addition, contact with cornicle secretions could also threaten an attacker and could provide direct fitness to the releaser. However, cornicle secretions may also be recruited as a kairomonal cue by aphid natural enemies. In this study, we investigated the effect of the cornicle droplet volatiles of the cabbage aphid, Brevicoryne brassicae (L.), on the host-searching behavior of naïve and experienced female Diaeretiella rapae (M(') Intosh) parasitoids in olfactometer studies. In addition, we evaluated the role of B. brassicae cornicle droplets on the oviposition preference of the parasitoid in a two-choice bioassay. Naïve females did not exhibit any preference between volatiles from aphids secreting cornicle droplets over non-secreting aphids, while experienced parasitoids exploited the secretions in their host location. Experienced females were also able to choose volatiles from both secreting and non-secreting aphids over clean air, while this ability was not observed in naïve females. Although secretion of cornicle droplets did not influence the percentage of first attack in either naïve or experienced females, the success of attack (i.e. resulting in a larva) was significantly different between secreting and non-secreting aphids in the case of experienced parasitoids.

Shallot aphids, Myzus ascalonicus, in strawberry: biocontrol potential of three predators and three parasitoids.

The parasitization capacity of 3 parasitoids and the predation capacity of 3 predators towards the shallot aphid, Myzus ascalonicus Doncaster (Homoptera: Aphididae), on strawberry, Fragaria x ananassa Duchesne (Rosales: Rosaceae) cv. Honeoye, were examined in laboratory experiments. In Petri dish assays, both Aphidius colemani Viereck (Hymenoptera: Aphidiidae) and A. ervi Haliday readily stung shallot aphids, with no significant difference in stinging frequency between the two species. A. ervi induced a significantly higher mortality (79.0 ± 7.2%) in terms of stung aphids compared with A. colemani (55.3 ± 4.1%); however, only a minor fraction (2.7 ± 1.8% and 7.1 ± 3.1%, respectively) of the killed aphids resulted in formation of mummies, presumably due to a physiological response to parasitism. The low percentage of mummification precludes the use of either Aphidius species in anything but inundative biocontrol. In similar set-ups, Aphelinus abdominalis (Dalman) (Hymenoptera: Aphelinidae) killed almost half (49.6 ± 5.3%) of the exposed aphids through host feeding. In addition, 23.2 ± 7.3% of non-host-fed aphids developed into mummified aphids, and 38.1 ± 13.2% of non-host-fed aphids died from other parasitoid-induced causes. However, the host feeding rate was reduced to only 1.2 ± 0.8%, and no significant parasitization mortality was observed on strawberry plants, suggesting that host plants interfered with A. abdominalis activity. This parasitoid does not, therefore, seem to be suited to either inoculative or inundative biocontrol of shallot aphids in strawberry. The three predators studied were the green lacewing, Chrysoperla carnea Steph. (Neuroptera: Chrysopidae), the two-spotted lady beetle, Adalia bipunctata L. (Coleoptera: Coccinellidae), and the gall midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae). Third instars of all 3 predators readily preyed upon the shallot aphid in Petri dish set-ups with significant differences in daily predation (34.62 ± 3.45, 25.25 ± 3.18, and 13.34 ± 1.45, respectively). Further studies on A. bipunctata revealed that the larvae maintained their daily predation capacity (32.0 ± 6.3) on strawberry plants. About 60% of already ovipositing A. bipunctata refrained from laying any eggs on the first day after transfer to set-ups with combinations of shallot or peach-potato aphids, Myzus persicae (Sulzer) (Homoptera: Aphididae), and strawberry or sweet pepper leaves. The aphid species and the plant species did not, however, have a significant influence on the number of females laying eggs, the average number of eggs laid during the first day being 6.37±1.28 per female. Adult lady beetles had a significant preference for odor from controls without plants over odors from uninfested strawberry or pepper plants, but they showed no preference for either of the plant species, whether infested with aphids or not. The predation capacity of A. bipunctata on shallot aphids holds promise for its use in inundative biocontrol, and the results on egg laying cues suggests that inoculative biocontrol may be possible, although further studies will be needed for a complete evaluation.

The green lacewing, Chrysoperla carnea: preference between lettuce aphids, Nasonovia ribisnigri, and Western flower thrips, Frankliniella occidentalis.

This study investigated the prey preference of 3(rd) instar green lacewing, Chrysoperla carnea Stephens (Neuroptera: Chrysopidae), between western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), and lettuce aphids, Nasonovia ribisnigri (Mosley) (Hemiptera: Aphididae) in laboratory experiments at 25 ± 1° C and 70 ± 5% RH with five prey ratios (10 aphids:80 thrips, 25 aphids:65 thrips, 45 aphids:45 thrips, 65 aphids:25 thrips, and 80 aphids:10 thrips). Third instar C. carnea larvae readily preyed upon both thrips and aphids, with thrips mortality varying between 40 and 90%, and aphid mortality between 52 and 98%. Chrysoperla carnea had a significant preference for N. ribisnigri at two ratios (10 aphids:80 thrips, 65 aphids:25 thrips), but no preference for either prey at the other ratios. There was no significant linear relationship between preference index and prey ratio, but a significant intercept of the linear regression indicated an overall preference of C. carnea for aphids with a value of 0.651 ± 0.054. The possible implications of these findings for control of N. ribisnigri and F. occidentalis by C. carnea are discussed.

Lethal and sublethal effects of fenpropathrin on the biological performance of Scolothrips longicornis (Thysanoptera: Thripidae).

Determination of negative nontarget effects of pesticides on beneficial organisms by measuring only lethal effects is likely to underestimate effects of sublethal doses. In this study, the sublethal effects of fenpropathrin on the predatory thrips Scolothrips longicornis Priesner (Thysanoptera: Thripidae) fed on Tetranychus urticae Koch (Acari: Tetranychidae) were evaluated under laboratory conditions. The estimated values of LC50 for female and male predators were 6.53 and 5.47 microg a.i./ml, respectively. Exposure to low-lethal concentrations (LC10, LC20, and LC30) of fenpropathrin significantly affected the biological characteristics of treated females of S. longicornis, the most noticeable effects being a shortening of female life span by > 70% accompanied by large reductions in oviposition period and fecundity. The offspring of females treated with low-lethal concentrations of fenpropathrin likewise had significantly reduced longevity, oviposition period, and fecundity, although not to the same extent as experienced by their mothers. Their juvenile development time was, however, not affected. These effects on the offspring were reflected in reduced rates of population increase and increased doubling times.

The effect of floral resources on parasitoid and host longevity: prospects for conservation biological control in strawberries.

The strawberry tortricid, Acleris comariana Lienig and Zeller (Lepidoptera: Tortricidae) is an important pest in Danish strawberry production. Its most common parasitoid is Copidosoma aretas (Walker) (Hymenoptera: Chalcidoidea: Encyrtidae). To identify selective flowering plants that could be used to increase functional biodiversity, the longevity of C. aretas and its host A. comariana was assessed on 5 flowering species: buckwheat, Fagopyrum esculentum Moench (Caryophyllales: Polygonaceae); borage, Borago officinalis L. (Boraginaceae); strawberry, Fragaria x ananassa Duchesne (Rosales: Rosaceae); phacelia, Phacelia tanacetifolia Bentham (Boraginaceae); and dill, Anethum graveolens L. (Apiales: Apiaceae). Dill was only tested with C. aretas. Sucrose and pollen served as positive controls, and pure water as a negative control. In a subsequent field experiment, A. comariana larval density was assessed at 1, 6, and 11 m distances from buckwheat flower strips in 3 fields. The proportion of field-collected larvae that were parasitized by C. aretas or fungi was assessed. Among the tested floral diets, buckwheat was superior for C. aretas, increasing its longevity by 1.4 times compared to water. Although buckwheat also increased longevity of A. comariana, its longevity and survival on buckwheat, borage, and strawberry was not significantly different, so buckwheat was chosen for field experiments. A. comariana densities in the 3 fields with sown buckwheat flower strips were 0.5, 4.0, and 8.3 larvae per m per row of strawberry respectively. Of the collected larvae, a total of 1%, 39%, and 65% were parasitized by C. aretas, respectively. The density of A. comariana and the proportion parasitized by C. aretas were highly significantly correlated. Distance from floral strips had no significant effect on either A. comariana larval density or on the proportion of individuals parasitized by C. aretas. Few other parasitoids emerged from collected larvae, and no larvae were infected by entomopathogenic fungi. Still, total A. comariana mortality was significantly affected by distance to flower strips, with the highest mortality near the flower strips. As no effect of buckwheat flower strips on C. aretas parasitism was found, the positive effect they had on A. comariana control stems from unknown mortality factors. As literature indicates that buckwheat for flower strips can augment a more complex suite of natural enemies, one such mortality factor could be a non-consumptive predator and/or parasitoid effect, but this requires further study. If confirmed, buckwheat may be utilized together with a selective food plant, once identified.

Effect of different temperatures on consumption of two spotted mite, Tetranychus urticae, eggs by the predatory thrips, Scolothrips longicornis.

Environmental variables such as temperature are important factors affecting the efficacy of biological control agents. This study evaluated the predation rate of the predatory thrips Scolothrips longicornis Priesner (Thysanoptera: Thripidae) against the two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae) under laboratory conditions. Based on daily and total prey consumption of different life stages of S. longicornis on spider mite eggs at temperatures covering the range suitable for development and survival of the predator (15° C to 37° C, 60 ± 10% RH, 16:8 L:D), there was a significant effect of temperature on prey consumption. The number of prey consumed daily by first and second instar larvae increased linearly with increasing temperature from 15 (°)C to 37 (°)C, whereas daily consumption of preovipositing and postovipositing females was uninfluenced by temperature. Lower temperature thresholds for consumption by first and second instar larvae of S. longicornis was estimated to be 6.8 ± 0.04° C and 4.6 ± 0.03° C, respectively. The daily consumption of ovipositing females followed a nonlinear pattern, with maximum daily predation estimated at 32.8° C. From the model used to describe consumption of ovipositing females, an upper threshold for consumption of 41.4° C was estimated. The performance of S. longicornis at the different temperatures is discussed in relation to its practical use in integrated pest control programs.

Effect of temperature on life table parameters of predatory thrips Scolothrips longicornis (Thysanoptera: Thripidae) fed on twospotted spider mites (Acari: Tetranychidae).

Environmental variables such as temperature are important factors that affect the efficiency of biological control agents. This study examined the effect of temperature on the sex ratio, longevity, oviposition periods, fecundity and life table parameters of the predatory thrips Scolothrips longicornis Priesner (Thysanoptera: Thripidae) fed on twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), at six constant temperatures: 15, 20, 26, 30, 35, and 37 degrees C. Approximately 75% of the progeny were female, except at 37 degrees C, at which temperature the proportion of males increased. Adult longevity as well as the preoviposition, oviposition, and postoviposition periods decreased significantly with temperature. Thus, adults lived for approximately 5 wk at 15 degrees C and < 1 wk at 37 degrees C with preoviposition, oviposition, and postoviposition periods ranging from 6.4 to 0.4, 24.4-3.1, and 7-0.8 d between the two temperature extremes, respectively. The maximum (56.48 eggs) and minimum (11.69 eggs) value of total fecundity was recorded at 26 and 37 degrees C, respectively. The intrinsic rate of natural increase (r(m)) of S. longicornis increased linearly with increasing temperature from 0.056 d(-1) at 15 degrees C to 0.310 d(-1) at 35 degrees C. The lower temperature threshold for the population increase of S. longicornis was estimated to be around 5 degrees C. The data suggest that the upper temperature threshold for the predatory thrips is approximately 37 degrees C. The results showed that populations of S. longicornis are able to develop at a broad range of temperatures and that this predator is well adapted to the high temperatures that occur in the Mediterranean region.

Prey preference of the predatory mite, Amblyseius swirskii between first instar western flower thrips Frankliniella occidentalis and nymphs of the twospotted spider mite Tetranychus urticae.

The prey preference of polyphagous predators plays an important role in suppressing different species of pest insects. In this study the prey preference of the predatory mite, Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) was examined between nymphs of the twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) and first instar larvae of the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), as well as between active and chrysalis spider mite protonymphs and active and chrysalis spider mite deutonymphs. The study was done in the laboratory on bean leaf discs at 25 ± 1° C and 70 ± 5% RH. Amblyseius swirskii had a clear preference for thrips compared to both spider mite protonymphs and deutonymphs. About twice as many thrips as spider mites were consumed. Amblyseius swirskii did not show a preference between active and chrysalis stages of spider mites.

Odour-mediated responses of a predatory mirid bug and its prey, the two-spotted spider mite.

It has been shown that many natural enemies of herbivorous arthropods use herbivore induced plant volatiles (HIPVs) to locate their prey. Herbivores can also exploit cues emitted by plants infested with heterospecifics or conspecifics. A study was conducted to test whether green bean HIPVs as well as odours emitted directly by spider mites influenced the orientation behaviour of the predatory mirid bug, Macrolophus caliginosus and its prey, Tetranychus urticae in a Y-tube olfactometer. Our results show that both spider mites and M. caliginosus preferred spider mite infested green bean plants to uninfested plants. For M. caliginosus this response was mediated by HIPVs whereas for T. urticae it was mediated through a composite response to both HIPVs and odours emitted directly by the conspecifics (and their associated products). The results may be of use in practical biocontrol situations, through e.g., plant breeding for improved HIPV production, conditioning of mass-reared predators to appropriate cues, and employment of "push-pull-strategies" by using HIPVs.