Prey stage preference of Amblyseius paraaerialis (Acari: Phytoseiidae) on varied life stages of the spider mites Tetranychus urticae, Tetranychus macfarlanei and Oligonychus biharensis (Acari: Tetranychidae) and exploring the mass rearing possibilities of this predatory mite on alternative diets.

Spider mites Tetranychus urticae, Tetranychus macfarlanei and Oligonychus biharensis are considered to be highly polyphagous in nature and causes severe damage to a wide range of plants around the world. Amblyseius paraaerialis is an efficient phytoseiid predator of spider mites with a potential to survive on both natural and alternative diets. Evaluation of predatory potential and prey stage preference provides valuable information on the efficacy of the predatory species in controlling mite population. Feeding experiments were conducted on mulberry leaf discs under the laboratory conditions of 30 ± 2 °C and 70 ± 5% relative humidity (RH). After 24 h of feeding experiment, the adult female predator exhibited a significant preference in feeding towards the eggs of T. macfarlanei (42.6%) and the larval stages of T. urticae (46%) and O. biharensis (25.3%). The mass rearing possibilities of A. paraaerialis was tested by tracking and comparing the developmental duration of individual life stages on varied food sources like, honey, castor (Ricinus communis) pollen, honey-pollen mixture and mixed life stages of T. urticae. The predator was failed to complete its development on honey and pollen when supplied separately. However it was successfully developed on honey-pollen mixture and mixed life stages of T. urticae. The developmental studies unravelled a shortest developmental duration and an extended adult longevity and lifespan of A. paraaerialis when reared on the alternative diet, thus opened up the mass rearing possibility of the predatory species under laboratory conditions.

Why are phytoseiid predatory mites not effectively controlling Echinothrips americanus?

The poinsettia thrips, Echinothrips americanus Morgan (Thysanoptera: Thripidae), is a key pest of various ornamental and vegetable greenhouse crops. As current biological control alternatives lack efficiency, applying chemicals remains the dominant control strategy, thereby heavily disturbing the biocontrol-based integrated management of other pests. For a range of other thrips pests, phytoseiid predatory mites have shown to be effective biocontrol agents, being able to overcome the thrips' physical and chemical defense armory. Here, we investigated potential underlying causes for the lack of phytoseiid efficacy in controlling E. americanus. First, we assessed the nutritional value of E. americanus for the predatory mite Amblydromalus limonicus (Garman and McGregor) (Acari: Phytoseiidae) when its physical or chemical defenses were eliminated by freezing the thrips. The phytoseiid could complete its immature development when frozen thrips instars were offered, but not when these were offered alive. Subsequently, we tested whether adult female A. limonicus had a higher predation rate on first instar E. americanus when they had been given experience with either live or frozen E. americanus during their immature development (i.e., conditioning). Conditioning significantly increased the predation capacity of the phytoseiid. Finally, we tested the control potential of conditioned A. limonicus versus naïve ones when exposed to E. americanus on sweet pepper plants. In contrast to the laboratory trials, at the plant level, conditioning did not yield better control. Possible factors explaining insufficient control of E. americanus by phytoseiids are discussed.

Expression and functional analysis of Niemann-Pick C2 gene in Phytoseiulus persimilis.

As a new protein class of semiochemical binding and transporting, Niemann-Pick proteins type C2 (NPC2) in arthropods have received more attentions in recent decade. However, the gene function has not been studied in phytoseiid mites with biocontrol potential. In the current study, we cloned a NPC2 gene PpNPC2a from the transcriptome of Phytoseiulus persimilis Athias-Henriot. By encoding 181 amino acids with a conserved ML domain, PpNPC2a was found a homolog of NPC2-1 in Galendromus occidentalis Nesbitt. We then measured the spatio-temporal expression of PpNPC2a in P. persimilis, and found the highest expression in female adults compared to other stages. Due to the tiny body size of predatory mites, we only examined tissue expressions in two sections: the anterior part (gnathosoma and the first pair of legs) and the posterior part (idiosoma without the first pair of legs). Higher transcription of PpNPC2a was found in the posterior part. To investigate the potential function of PpNPC2a in P. persimilis, we interfered gene expression in female adults by feeding dsRNA, which resulted in a decrease of relative expression by 59.1 and 78.2% after 24 and 72 h, respectively. Compared with the control, dsNPC2a-treated P. persimilis were insensitive to the scent of leaves or plants infested by spider mites, suggesting a role of PpNPC2a in response to plant volatiles. However, the dsNPC2a-interfered mites could still respond to four representative compounds of herbivore-induced plant volatiles, including 4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT), methyl salicylate (MeSA), ß-caryophyllene and linalool. In short, our results indicated PpNPC2a may be involved in the chemosensory process of P. persimilis in response to whole-plant volatiles.

Amblyseius orientalis shows high consumption and reproduction on Polyphagotarsonemus latus in China.

The broad mite, Polyphagotarsonemus latus (Banks) (Acari: Tarsonemidae), is a cosmopolitan pest that infests many greenhouse crops. Biological control is an important way to control P. latus, with predatory mites being the most widely used natural enemies of this pest. The purpose of this study is to evaluate the capabilities of three native phytoseiids in China (Neoseiulus californicus, Neoseiulus barkeri and Amblyseius orientalis) in controlling P. latus, using Amblyseius swirskii as a control, a commercial biocontrol agent of this pest widely used in Europe. Consumption, development, and reproduction of the four species when fed with P. latus were assessed, and their life table parameters were estimated and compared. Among the three native species, A. orientalis has the highest consumption rate of P. latus (29.0 per day), the shortest developmental duration (5.3 days), and the highest cumulative fecundity (13.5 eggs/female). Overall, its intrinsic rate of increase (rm) is 0.12, comparable to that of A. swirskii. Among the three candidates, A. orientalis is the only one whose population increase might be expected when fed with P. latus. Therefore, we propose A. orientalis to be a potential biocontrol agent for this pest in China.

Expression and functional analysis of transformer-2 in Phytoseiulus persimilis and other genes potentially participating in reproductive regulation.

Transformer-2 (tra-2) is an important sex-determining gene in insects. It also plays a role in the reproduction of phytoseiid mites. We performed bioinformatic analyses for the tra-2 ortholog in Phytoseiulus persimilis (termed Pptra-2), measured its expression at different stages and quantitatively identified its function in reproduction. This gene encodes 288 amino acids with a conserved RRM domain. The peak of its expression was observed in adult females, especially ca. 5 days after mating. In addition, expression is also higher in eggs than in other stages and adult males. When Pptra-2 was silenced through RNA interference with oral delivery of dsRNA, 56% of the females had their egg hatching rates decreased in the first 5 days, from ca. 100% to ca. 20%, and maintained at low levels during the rest of the oviposition period. To detect other genes functionally related to Pptra-2, transcriptome analyses were performed on day 5 after mating. We compared mRNA expressions among interfered females with significantly reduced egg hatching rate, interfered females without significant hatching rate and CK. In total 403 differential genes were identified, of which 42 functional genes involved in the regulation of female reproduction and embryonic development were screened and discussed.

Pollen Alone or a Mixture of Pollen Types? Assessing Their Suitability for Mass Rearing of Neoseiulus cucumeris (Acari: Phytoseiidae) Over 20 Generations.

The generalist predatory mite, Neoseiulus cucumeris (Oudemans) is known as one of the most effective natural enemies on many pests. This economically important biocontrol agent was reared for 20 generations on date palm and castor bean pollen as well as a mixture of pollen types, including date palm, castor bean, and almond. The performance of this predator was evaluated by comparing its life table parameters after different generations (G1-G20) fed on each diet in a laboratory at 25 ± 1°C, 60 ± 5% RH, and a photoperiod of 16:8 (L: D) h. The development time of the predator reared on all tested diets had no significant difference in G20. The intrinsic rate of increase (r) of N. cucumeris by feeding on the mixed pollen (0.197 day-1) was significantly higher than that on castor bean, or date palm pollen (0.146 or 0.152 day-1 in G1, respectively). Our results indicated that the predator's performance was not affected by long-term feeding on the pollen diets, as well as there was no considerable difference between pollen alone and pollen mixture diets. Furthermore, mites reared on pollen diets had higher quality than those reared on natural prey, Tetranychus urticae Koch.

Molecular characterization, expression, and function of Vitellogenin genes in Phytoseiulus persimilis.

Vitellogenin (Vg) is an important factor that impacts oocyte maturation, egg formation and embryonic development in Arthropoda. Two orthologs of Vg gene were obtained from the genome of Phytoseiulus persimilis and termed as PpVg1 and PpVg2. Both orthologs belong to the large lipid transfer protein superfamily. The expression of PpVg1 and PpVg2 was low in immatures and male adults, and increased rapidly in female adults after mating, and reached a peak before the first egg was laid (168×  and 20.5×  the level in virgin females, respectively). When PpVg1 and PpVg2 were interfered with dsRNA, the relative expression decreased by 81.0 and 30.9%, respectively, and 7.8 and 31.4% interfered individuals died within 24 h. Among surviving individuals, ca. 51.1 and 44.8% are infertile. Factors that might be related to expression of Vg genes are also discussed.

Population dynamics of mites in slow-release sachets used in biological control: a new study methodology.

Predatory mite species (Acari: Phytoseiidae) are essential tools in the biological control of greenhouses pests. The natural enemies can be released directly into a crop. A better, partly preventive system is to place slow-release sachets on the plants. Inside such sachets is a factitious prey's food substrate-which also acts as refuge-and the predator. The objective of this study was to develop a new methodology to evaluate the population dynamics of this sachet system, based on the factitious prey Carpoglyphus lactis and the predatory mite Amblyseius swirskii. Through two tests carried out under laboratory conditions, the sachets were first compared to the traditional extraction method that uses Berlese-Tullgren funnels and an extraction method using flotation in hexane. The latter method proved more effective at sampling the motile states (larvae, nymphs, and adults), both for the predatory species and for the factitious prey, extracting up to 3.7 × more mites than the Berlese-Tullgren funnel. Second, the population dynamics of both mite species was studied in a laboratory test, both inside and outside the sachets. In this way, a positive correlation was demonstrated between the number of predatory mites and the number of prey mites inside the sachets. Conversely, no correlation was found between the interior population of predatory mites and the number that venture outside. We can conclude that hexane extraction is very useful both in quality control of predatory mites and in studying how the sachets behave when faced with various factors.

Review: predatory soil mites as biocontrol agents of above- and below-ground plant pests.

Biological pest control is becoming increasingly important for sustainable agriculture. Although many species of natural enemies are already being used commercially, efficient biological control of various pests is still lacking, and there is a need for more biocontrol agents. In this review, we focus on predatory soil mites, their role as natural enemies, and their biocontrol potential, mainly in vegetable and ornamental crops, with an emphasis on greenhouse systems. These predators are still underrepresented in biological control, but have several advantages compared to predators living on above-ground plant parts. For example, predatory soil mites are often easy and affordable to mass rear, as most of them are generalist predators, which also means that they may be used against various pests and can survive periods of pest scarcity by feeding on alternative prey or food. Many of them can also endure unfavourable conditions, making it easier for them to establish in various crops. Based on the current literature, we show that they have potential to control a variety of pests, both in greenhouses and in the field. However, more research is needed to fully understand and appreciate their potential as biocontrol agents. We review and discuss several methods to increase their efficiency, such as supplying them with alternative food and changing soil/litter structure to enable persistence of their populations. We conclude that predatory soil mites deserve more attention in future studies to increase their application in agricultural crops.

Arthropods associated with the lychee erinose mite, Aceria litchii (Acari: Eriophyidae) on lychee trees in Minas Gerais, Brazil.

Lychee fruit production (Litchi chinensis) has been threatened in many regions of the world where the presence has been reported of the litchii erinose mite, Aceria litchii (Keifer). This study aims to identify the arthropod community associated with A. litchii on lychee plants in Minas Gerais, Brazil, and to seek for potential natural enemies associated with this mite pest in this region. We sampled lychee leaves infested with A. litchii in commercial and non-commercial lychee orchards during seven consecutive months, covering the dry and wet seasons of the year. Arthropods found in association with A. litchii on lychee leaves were collected and identified. The results indicate that a great diversity and abundance of mites and hexapods are associated with A. litchii. A total of 985 and 1872 specimens of mites were identified in commercial and non-commercial areas, respectively, belonging to the families Cheyletidae, Cunaxidae, Eupodidae, Iolinidae, Stigmaeidae, Phytoseiidae, Tarsonemidae, Tenuipalpidae, Tetranychidae, Tryophtydeidae, Tuckerellidae, Tydeidae, Winterschmidtiidae and Xenocaligonellidae and the suborder Oribatida. Among them, Phytoseiidae was the most abundant and diverse family with a total of 11 species identified, in which Phytoseius intermedius was the most abundant predatory mite species collected. Minor specimens of hexapods were also been, belonging to the orders Collembola, Blattodea, Coleoptera, Diptera, Hemiptera, Hymenoptera, Psocoptera and Thysanoptera. The presence of a high community of predatory mites in association with A. litchii deserves attention and our results indicate that studies to test the potential of these species and the adoption of management practices that enhance this ecological service must be carried out to achieve satisfactory control of the lychee erinose mite in lychee plants.

Effects of SmartWater, a fluorescent mark, on the dispersal, behavior, and biocontrol efficacy of Phytoseiulus persimilis.

New marking methods for studying small biocontrol agents (especially predatory mites) are needed because many current techniques are expensive, ineffective or not applicable to small organisms. The objective of this study was to determine whether SmartWater, a liquid and permanent fluorescent dye, can be used to mark Phytoseiulus persimilis for experimentation without any deleterious effects on its dispersal, behavior, reproduction, and biocontrol efficacy. Our results show that there were no significant differences in movement, inter-plant dispersal, feeding behavior, survivability, and reproduction between marked P. persimilis and control individuals sprayed with water. We also found that the SmartWater mark lasted for the duration of the mites' life, indicating strong durability over time. Marking efficacy may be reduced, due to a trade-off between batch marking efficacy and the possibility of drowning study organisms. However, we feel future research could improve liquid marking techniques that would reduce this risk. Overall, this study concludes that SmartWater could be a useful marking tool for predatory mites in both laboratory and field studies.

Molecular variation of the cytochrome b DNA and protein sequences in Phytoseiulus macropilis and P. persimilis (Acari: Phytoseiidae) reflect population differentiation.

Several phytoseiid mite species are important natural enemies used in biological control strategies. In the present study, Cytb mtDNA sequences of various populations of two species, Phytoseiulus macropolis and P. persimilis, were compared to determine whether the specimens collected in Brazil could belong to P. persimilis as this latter species is reported in South America but not in Brazil. The Cytb marker was used because of its high evolution rate, assumed to capture intraspecific variation. No overlap between intra- and interspecific distances was observed but the distances were quite low for interspecific variation. This can be due to the particular biology of Phytoseiulus species and this shows the difficulty to apply a universal threshold in genetic distances to conclude about the existence of one or several species. Cytb mtDNA sequences were also considered to assess intraspecific variation. The DNA sequences of P. persimilis populations were very similar, probably because they all originated from the West Palearctic region or because of a prevalence of commercialized specimens in natura. For P. macropilis, higher genetic distances were observed and differentiation was noted according to geographic location and, to a smaller extent, pyrethroid resistance. To determine how DNA variation might impact the protein function (CytB fragment considered), the amino acid compositions of the populations studied were compared. No diagnostic mutation was observed between pyrethroid resistant and susceptible populations, whereas four mutations were identified between populations of P. macropilis separated by 1300 km (different climatic conditions). The impact of such mutations is discussed but knowledge is scarce, which makes it difficult to root testable hypotheses. The protein analysis clearly opens new perspectives in Phytoseiidae studies.

Plant-feeding may explain why the generalist predator Euseius stipulatus does better on less defended citrus plants but Tetranychus-specialists Neoseiulus californicus and Phytoseiulus persimilis do not.

The generalist predator Euseius stipulatus (Athias-Henriot) and the Tetranychidae-specialist predators Neoseiulus californicus (McGregor) and Phytoseiulus persimilis Athias-Henriot play a key role in the regulation of Tetranychus urticae Koch in Spanish citrus orchards. Previous studies have shown that sour orange (Citrus aurantium L.) and Cleopatra mandarin (Citrus reshni hort. ex Tan.) display extreme resistance and susceptibility to T. urticae, respectively. When offered a choice between these two genotypes infested by T. urticae, E. stipulatus preferred Cleopatra mandarin, whereas the specialists did not show any preference. The present study was undertaken to check whether these preferences could be related to the feeding of E. stipulatus on the host plant and/or to differences in prey feeding on the two plants. Our results demonstrate that E. stipulatus is a zoophytophagous mite, which can engage in direct plant feeding in sour orange and Cleopatra mandarin, whereas neither N. californicus nor P. persimilis do so. Whereas Cleopatra mandarin provided a higher-quality prey/feeding substrate for E. stipulatus, which may be related to its phytophagy, no differences were observed for the two specialists. As higher constitutive and faster inducible defense against T. urticae in sour orange relative to Cleopatra mandarin plants result in sour orange supporting lower T. urticae densities and plant damage, our results demonstrate that pest regulation by specialist natural enemies may be more effective when prey feed on better defended plants.

Impact of a tarsonemid prey mite and its fungal diet on the reproductive performance of a predatory mite.

Phytoseiid predatory mites are the most important group of biocontrol agents currently used in protected cultivations worldwide. The possibility to produce these predators at high densities on factitious prey mites is a crucial factor for their success. Commonly used factitious prey mites comprise mainly species belonging to the cohort of Astigmatina. In the present study, we investigated the potential of tarsonemid prey mites as a food source for the spider mite predator Neoseiulus californicus (McGregor) (Acari: Phytoseiidae). The oviposition of N. californicus on mixed stages of Tarsonemus fusarii Cooreman (Acari: Tarsonemidae) was similar to that on its natural prey, the two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae). As most tarsonemids are specialized fungus-feeders, we tested the effect of different fungal species on the growth of T. fusarii. Subsequently, we analysed the impact on the fungal growing medium on the oviposition of N. californicus. The fungal growing medium of T. fusarii had a significant negative effect on the reproductive output of the predatory mite. When T. fusarii was separated from the rearing medium, these detrimental effects were not observed. The present study shows the potential of using tarsonemid prey mites in the production of phytoseiid predatory mites.

Maternal effect determines drought resistance of eggs in the predatory mite Phytoseiulus persimilis.

The ability of an organism to adapt to short-term environmental changes within its lifetime is of fundamental importance. This adaptation may occur through phenotypic plasticity. Insects and mites, in particular, are sensitive to changes in temperature and humidity, especially during the juvenile stages. We studied the role of phenotypic plasticity in the adaptation of eggs to different relative humidity conditions, in the predatory mite Phytoseiulus persimilis, used worldwide as a biological control agent of the spider mite Tetranychus urticae. The biocontrol efficacy of P. persimilis decreases under dry conditions, partly because P. persimilis eggs are sensitive to drought. We exposed P. persimilis adult females from two different strains to constant and variable humidity regimes and evaluated the hatching rate of their eggs in dry conditions, as well as the survival and oviposition rates of these females. Whereas the eggs laid by P. persimilis females exposed to constant high humidity did not survive in dry conditions, females exposed to constant low humidity started laying drought-resistant eggs after 24 h of exposure. Survival and oviposition rates of the females were affected by humidity: females laid fewer eggs under constant low humidity and had a shorter lifespan under constant high and constant low humidity. The humidity regimes tested had similar effects across the two P. persimilis strains. Our results demonstrate that transgenerational phenotypic plasticity, called maternal effect, allows P. persimilis females to prepare their offspring for dry conditions.

Does feeding on pollen grains affect the performance of Amblyseius swirskii (Acari: Phytoseiidae) during subsequent generations?

Diet is a critical component of the mass-rearing of biological control agents, but the impacts of diet are not always immediately obvious and can take several generations to manifest, resulting in poor survival, reproduction, and ability to kill prey under natural conditions. Our present study aimed to investigate the performance of a commercially-reared phytoseiid mite, Amblyseius swirskii, after four (G4) and six (G6) consecutive generations on pollen grains of two plant species, as well as its ability to find and kill its natural prey, Tetranychus urticae, after long-term rearing on each diet. We found no significant difference between the two diets in intrinsic and finite rates in G4. However, both diet and generation exerted a significant influence on the fecundity of A. swirskii. By G6, females reared on almond pollen had greater net reproductive and intrinsic rate compared to those reared on maize pollen. Conversely, A. swirskii fed on maize pollen consumed fewer prey than those reared on other diets, especially at higher prey densities. The findings have important implications for developing the mass-rearing program of A. swirskii on non-prey diets. Further research must explore the suitability of almond pollen in the large-scale culture of A. swirskii.

Comparison of the phytoseiid mites Amblyseius swirskii and Amblydromalus limonicus for biological control of chilli thrips, Scirtothrips dorsalis (Thysanoptera: Thripidae).

The chilli thrips, Scirtothrips dorsalis Hood, is a recently established pest in the USA and poses a serious risk to many economically important ornamental and food crops. In this study the biological control potential of the phytoseiid mites Amblydromalus limonicus (Garman and McGregor) and Amblyseius swirskii (Athias-Henriot) was compared by examining their predation and oviposition rates when fed different developmental stages of S. dorsalis. Gravid females were offered 10 individuals of either first instar, second instar, or adult S. dorsalis using a no-choice leaf disc bioassay and oviposition and predation rates were assessed daily for 2 and 3 days, respectively. There was no significant difference in predation and oviposition rates between mite species fed specific S. dorsalis life stages. There was, however, a significant effect of S. dorsalis life stage on the oviposition and predation rates observed for each mite species. The larval stage was the most preferred stage for both mite species, with A. swirskii consuming 4.6-6.3 and A. limonicus 4.8-6.4 individuals/day compared to only 1.6-1.7 adults/day consumed by both species. Female A. swirskii and A. limonicus laid 0.55-0.75 and 0.73 eggs/day on the two larval stages, respectively, compared to only 0.25-0.30 eggs/day observed for individuals feeding on adults. Although the results showed that the biological control potential of both mite species was similar, having an additional predator available that may be as effective as A. swirskii, a proven control agent against S. dorsalis in the field, warrants additional research into its potential utility.

Microscopic analysis of the microbiota of three commercial Phytoseiidae species (Acari: Mesostigmata).

Microbes associated with the external and internal anatomy of three commercially available predatory mite species-Phytoseiulus persimilis, Typhlodromips (= Amblyseius) swirskii, and Neoseiulus (= Amblyseius) cucumeris-were examined using light microscopy, confocal laser scanning microscopy and fluorescence in-situ hybridization (FISH). Four microbe morphotypes were observed on external body regions. These included three microfungi-like organisms (named T1, T2 and T3) and rod-shaped bacteria (T4). Morphotypes showed unique distributions on the external body regions and certain microbes were found only on one host species. Microfungi-like T1 were present in all three species whereas T2 and T3 were present in only P. persimilis and T. swirskii, respectively. T1 and T2 microbes were most abundant on the ventral structures of the idiosoma and legs, most frequently associated with coxae, coxal folds, ventrianal shields and epigynal shields. T3 microbes were most abundant on legs and dorsal idiosoma. T4 microbes were less abundant and were attached to epigynal shields of N. cucumeris and T. swirskii. Significant differences in distribution between batches suggest temporal fluctuations in the microbiota of phytoseiids in mass-reared systems. FISH showed bacteria within the alimentary tract, in Malpighian tubules and anal atria. These may aid absorption of excretory products or maintaining gut physiology. We suggest a mechanism by which microbes may be transmitted to offspring and throughout populations. This study aims to improve our knowledge of this poorly understood area and highlights the necessity of understanding the microbiota of Acari.

Molecular characterization of phytoseiid mites in Turkey based on the internal transcribed spacer (ITS) region, with a new record for the country.

The family Phytoseiidae contains many predatory mite species and some are used in biological control programs worldwide. The identification of phytoseiid mites is based on tiny morphological structures and sometimes species diagnosis is not easy especially for non-taxonomists. DNA-based approaches may offer a fast and accurate diagnosis to overcome these difficulties, nevertheless more DNA sequences are needed to determine intra- and inter-specific variations and to provide accurate decision rules based on genetic distances between the taxa considered. In this study, we provide the molecular characterization of seven phytoseiid species based on the internal transcribed spacer (ITS) region. Several populations of these species collected in Turkey were considered. A phylogenetic tree was also constructed. Finally, we record the presence of Neoseiulus reductus (Wainstein) in Turkey.

Compatibility of two predator species for biological control of the two-spotted spider mite.

Because predators may interfere with each other, an important step towards the implementation of successful release of multiple predators in biocontrol programs requires resolving how predators respond to the presence of heterospecific competitors. Several species of predatory mites are important biocontrol agents and the species Phytoseiulus macropilis and Neoseiulus californicus are used to control the two-spotted spider mite, Tetranychus urticae, an important pest in agriculture worldwide. We investigated their compatibility showing that the two predators do not avoid plants on which the other species is present together with their common prey, and demonstrated that their oviposition rates are not affected by the presence of the other species. However, the distribution of the eggs on leaf discs was affected by the presence of the heterospecific predator. This behaviour might weaken possible interference between these two biocontrol agents, which, in turn, may enable their persistence on plants and favour pest suppression. The increased joint use of several natural enemies for biological control highlights the importance of studies on predator-predator interactions.