Effectiveness of different release rates of Trichogramma cacoeciae (Hymenoptera: Trichogrammatidae) against Tuta absoluta (Lepidoptera: Gelechiidae) in protected and open field tomato crops in Tunisia

Tuta absoluta (Meyrick) is a key insect pest of tomato crop causing major economic losses worldwide. Biological control using Trichogramma parasitoids is considered a promising, eco-friendly management tactic of this pest. We performed inundative releases of Trichogramma cacoeciae (Marchal) (Hymenoptera: Trichogrammatidae) against T. absoluta in Takelsa (northeastern Tunisia). Three weekly releases of either (i) 10, 20 or 30 Trichogramma per plant in protected (greenhouse) tomato crops or (ii) 20 or 40 Trichogramma per plant in open field tomatoes were evaluated for their effectiveness. Results indicated that 20 Trichogramma per plant was the most effective release rate in significantly decreasing the pest’s life stage densities. Parasitism rates were significantly higher on apical tomato leaves (estimated at 58.6% for the rate 20 Trichogramma per plant in greenhouses and at about 56% for the rate 40 Trichogramma per plant in open field) compared to those observed on middle leaves (24.6% and 45.26% for rate 2 respectively under greenhouse and open field conditions). Furthermore, the overall number of T. absolua eggs and larvae was significantly reduced after releases of Trichogramma parasitoids. These results clearly indicate that T. cacoeciae could be a promising biocontrol agent of T. absoluta in the largest tomato-producing area in Tunisia.     

Efficacy of entomopathogenic nematodes against the tomato leafminer <Emphasis Type="Italic">Tuta absoluta</Emphasis> in laboratory and greenhouse conditions

The tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is an important pest of tomato crops in South America and it has recently arrived in Europe affecting tomato plantations. The susceptibility of T. absoluta larvae and pupae to three species of entomopathogenic nematodes (Steinernema carpocapsae, Steinernema feltiae and Heterorhabditis bacteriophora) was examined under laboratory conditions. Leaf bioassays were conducted to evaluate the nematode’s capability to reach the larvae and kill them within the galleries. The efficacy of the three nematode species after foliar application to potted tomato plants was evaluated under greenhouse conditions. High larval mortality (78.6–100%) and low pupae mortality (<10%) was determined in laboratory experiments. In the leaf bioassay a high level of larval parasitation (77.1–91.7%) was recorded revealing the nematode’s capacity to kill the larvae inside the galleries. In the pot experiments nematode treatment reduced insect infection of tomato plants by 87–95%. The results demonstrate the suitability of entomopathogenic nematodes for controlling T. absoluta.     

Role of larval host plant experience and solanaceous plant volatile emissions in Tuta absoluta (Lepidoptera: Gelechiidae) host finding behavior

The tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae), is considered to be a major pest that damages tomato (Solanum lycopersicum L; Solanaceae) crops in South American, European, and Mediterranean countries. This insect species is polyphagous (i.e., feeds on many types of food); hence, it could also develop on other cultivated host plants, principally solanaceous plants, such as potato (S. tuberosum L.; Solanaceae) and eggplant (S. melongena L.; Solanaceae). Therefore, we tested the hypothesis that host plant choice by adult T. absoluta is influenced by plant volatile organic compounds and larval host plant experience. One tomato cultivar (cv.) ‘Moneymaker’ and three potato cv. ‘Charlotte’ ‘Bintje,’ and ‘Nicola’ were tested. Using a flying tunnel, we observed that females reared on tomato preferred flying toward tomato and, to a lesser extent, potato cv. ‘Charlotte.’ These preferences might be explained by the high release of terpenes by these two plants. When conducting oviposition choice assays, we found no preference between tomato and potato in the number of eggs laid by females that had been previously reared on either host plant. This study indicates that the host finding behavior of T. absoluta is mediated by solanaceous volatiles, while oviposition behavior appears to depend on additional stimuli. These results provide baseline information for use in the development of new control strategies against T. absoluta using semiochemicals and plant breeding.     

Native parasitoids associated with Tuta absoluta in the tomato production areas of the Spanish Mediterranean Coast

The tomato borer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is an invasive pest that produces significant damage to tomato crops in the Mediterranean area. Although several species of predatory bugs are successfully being used for biological control of the pest, little is known about the parasitoids that are able to exploit T. absoluta as a host. With the aim of better understanding parasitoid species richness of T. absoluta along the Mediterranean Spanish Coast, we conducted an extensive survey to determine distribution, host plants and habitats where parasitoids are present. Our results indicated that egg parasitoids are naturally scarce but that the species richness of larval/pupal parasitoids is high and includes 20 different species. Seven of these had not been previously reported as T. absoluta parasitoids. The most frequent parasitoid species recovered were Necremnus sp. nr. artynes (Walker), Stenomesius cf. japonicus (Ashmead) and Neochrysocharis formosa (Westwood) (Hymenoptera: Eulophidae).     

Sharing a predator: can an invasive alien pest affect the predation on a local pest?

Invasive species can strongly affect biotic interactions in ecosystems, interacting both directly and indirectly with local species. In European tomato greenhouses, the invasive alien pest Tuta absoluta may impact the population dynamics of other pests like whiteflies. Besides inducing damages to the host plant and competing for resources with local pests, this alien species may exert a predator-mediated interaction on local pests sharing common natural enemies. Biocontrol agents usually used against whiteflies may also prey upon T. absoluta and this could alter the dynamics of local pest populations. We evaluated possible resource competition and predator-mediated interactions in a system involving one mirid predator Macrolophus pygmaeus and two pests, T. absoluta and a local whitefly, Bemisia tabaci, on greenhouse tomatoes. Results showed that both resource competition and predator-mediated interactions occurred simultaneously. In the presence of the shared predator, there was a short-term positive effect of T. absoluta on B. tabaci [up to 5.9-fold increase of B. tabaci juveniles (egg + larvae) after four weeks]. However, in the long-term there was a negative predator-mediated interaction of T. absoluta on B. tabaci, i.e., after ten weeks the density of B. tabaci was 7.3-fold lower in the presence of the invasive pest. We emphasize the critical role of generalist predators in managing both local and invasive alien pest populations and that the strength and direction of predator-mediated indirect interactions can depend on the time scale considered.     

Modeling the risk of invasion and spread of Tuta absoluta in Africa

Tuta absoluta is an invasive insect that originated from South America and has spread to Europe Africa and Asia. Since its detection in Spain in 2006, the pest is continuing to expand its geographical range, including the recent detection in several Sub-Saharan African countries. The present study proposed a model based on cellular automata to predict year-to-year the risk of the invasion and spread of T. absoluta across Africa. Using, land vegetation cover, temperature, relative humidity and yield of tomato production as key driving factors, we were able to mimic the spreading behavior of the pest, and to understand the role that each of these factors play in the process of propagation of invasion. Simulations by inferring the pest’s natural ability to fly long distance revealed that T. absoluta could reach South of Africa ten years after being detected in Spain (Europe). Findings also reveal that relative humidity and the presence of T. absoluta host plants are important factors for improving the accuracy of the prediction. The study aims to inform stakeholders in plant health, plant quarantine, and pest management on the risks that T. absoluta may cause at local, regional and event global scales. It is suggested that adequate measures should be put in place to stop, control and contain the process used by this pest to expand its range.     

New records of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) predation by Brazilian Hemipteran predatory bugs

The tomato borer Tuta absoluta, native to western South America, is an extremely devastating pest in tomato crops in most of South America, Europe and Africa North of the Sahel, causes yield losses up to 100% and decreases fruit quality in open field and greenhouse crops if control methods are not applied. In Brazil two other important lepidopteran pests – Neoleucinodes elegantalis and Helicoverpa zea – occur in tomato, as well as thrips, whiteflies and aphids. For control of these pests, frequent applications of pesticides of up to 5 times per week are needed, and these resulted in the appearance of resistant populations to a number of active ingredients and decimation of natural enemies. Biological control may offer a better, safer and more sustainable opportunity for pest management. Mirid predatory bugs are currently used with success in southern Europe to control T. absoluta and other pests. In Brazil, four Hemipteran predatory bugs, not yet known to attack T. absoluta, were found to successfully prey on eggs and larvae of this pest. The first results on their predation capacity, development, survival and reproduction on T. absoluta on tomato plants are presented.     

Effects of host-egg ages on host selection and suitability of four Chinese Trichogramma species,egg parasitoids of the rice striped stem borer,Chilo suppressalis

The striped stem borer, Chilo suppressalis (Walker), is one of the most economically important rice pests worldwide. However, biological control of this pest using natural-enemy insects has been rarely documented to date. With the objective of screening suitable candidate species for controlling the striped stem borer, we investigated the effect of the age of host eggs on the host selection and suitability by four indigenous Trichogramma species on their native host, C. suppressalis. The results indicated that the differently aged eggs of C. suppressalis were all accepted by T. japonicum, T. dendrolimi and T. chilonis, and there was a clear tendency to parasitize older eggs less under no-choice and choice conditions. The number of parasitized host eggs by T. ostriniae also decreased with the increasing host age in the no-choice test, but more 2-day-old host eggs were parasitized in the choice test. When 0-, 2-day-old eggs were offered, T. dendrolimi, T. japonicum and T. chilonis exhibited similar parasitism ability, whereas T. ostriniae appeared to have a stronger ability to attack the older host eggs (4-day-old). Trichogramma japonicum developed and emerged on parasitized C. suppressalis eggs of all ages tested, while showing a better adaptation to younger host eggs with significantly faster developmental time, higher survival and more female progeny on 0-day-old eggs. No adults for each of the other three Trichogramma species emerged from parasitized 4-day-old host eggs, and they had similar developmental time, survival and female progeny on parasitized 0-, 2-day-old host eggs with an exception of female progeny for T. chilonis. On 0-day-old host eggs, T. japonicum developed faster and T. ostriniae had lower progeny survival than the other three Trichogramma species evaluated, respectively. The current study provides useful information to select suitable Trichogramma species for controlling the striped stem borer, C. suppressalis.     

Molecular and morphological diagnoses of five species of Trichogramma: biological control agents of Chrysodeixis chalcites (Lepidoptera: Noctuidae) and Tuta absoluta (Lepidoptera: Gelechiidae) in the Canary Islands

Prospecting for potential natural enemies of the invasive lepidopteran tomato pest Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) and the banana pest Chrysodeixis chalcites (Esper) (Lepidoptera: Noctuidae) on the Canary Islands archipelago, where no Trichogramma species were previously recorded, has led to the discovery of five distinct species. T. achaeae Nagaraja & Nagarkatti, T. bourarachae Pintureau & Babault, T. euproctidis (Girault) and T. evanescens Westwood are relatively widespread species. The fifth is close to T. brassicae Bezdenko, but differs sufficiently in the sequence of the ITS2 region of ribosomal RNA to warrant further investigation as a species probably new to science. Each species is treated in detail in order to facilitate identification in future using molecular and/or morphological characters, or a combination of both. All species are newly recorded for the Canary Islands, and the distribution of each within the islands and elsewhere is provided. Known host records are given within the Canary Islands and elsewhere. The most common species found, T. achaeae, is already being used in biological control programmes against T. absoluta in mainland Spain and field trials are ongoing to evaluate its effectiveness as a biological control agent of C. chalcites in banana crops.     

Preference and Prey Switching in a Generalist Predator Attacking Local and Invasive Alien Pests

Invasive pest species may strongly affect biotic interactions in agro-ecosystems. The ability of generalist predators to prey on new invasive pests may result in drastic changes in the population dynamics of local pest species owing to predator-mediated indirect interactions among prey. On a short time scale, the nature and strength of such indirect interactions depend largely on preferences between prey and on predator behavior patterns. Under laboratory conditions we evaluated the prey preference of the generalist predator Macrolophus pygmaeus Rambur (Heteroptera: Miridae) when it encounters simultaneously the local tomato pest Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) and the invasive alien pest Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae). We tested various ratios of local vs. alien prey numbers, measuring switching by the predator from one prey to the other, and assessing what conditions (e.g. prey species abundance and prey development stage) may favor such prey switching. The total predation activity of M. pygmaeus was affected by the presence of T. absoluta in the prey complex with an opposite effect when comparing adult and juvenile predators. The predator showed similar preference toward T. absoluta eggs and B. tabaci nymphs, but T. absoluta larvae were clearly less attacked. However, prey preference strongly depended on prey relative abundance with a disproportionately high predation on the most abundant prey and disproportionately low predation on the rarest prey. Together with the findings of a recent companion study (Bompard et al. 2013, Population Ecology), the insight obtained on M. pygmaeus prey switching may be useful for Integrated Pest Management in tomato crops, notably for optimal simultaneous management of B. tabaci and T. absoluta, which very frequently co-occur on tomato.     

Efficacy of sulphur on Tuta absoluta and its side effects on the predator Nesidiocoris tenuis

The South American tomato pinworm, Tuta absoluta (Meyrick), is one of the major pests of tomato crop. Since its detection in the Mediterranean basin, it has been commonly controlled using chemical insecticides. However, inoculation and conservation of predatory mirids, integrated with sprays of selective insecticides, has been demonstrated to be a cost‐effective strategy for controlling this pest. In this work, we tested the efficacy of two sulphur formulations, dustable and wettable powder, for controlling T. absoluta on tomato under greenhouse and open‐field conditions. In addition, the side effects of both sulphur formulations on the predator, Nesidiocoris tenuis (Reuter), were evaluated under laboratory conditions. Dustable sulphur, applied weekly on tomato seedlings artificially infested with T. absoluta in greenhouse conditions, significantly reduced the infestation levels and was demonstrated to have a repellent effect on oviposition. Wettable sulphur was not effective for controlling T. absoluta populations in both greenhouse and open‐field experiments. In the side effect trials conducted with N. tenuis, only dustable sulphur resulted in being moderately harmful as a fresh residue and slightly harmful as a 7‐day‐old residue; no effects were recorded exposing the predator to 14‐day‐old sulphur residues. In contrast, wettable sulphur was classified as harmless to N. tenuis. Our results suggest that the use of sulphur, especially as dustable powder, could be considered as a tool in T. absoluta management strategies, although its side effects on N. tenuis should be taken into account. The implications of these results for the use of sulphur formulations in pest and disease management programmes in tomato crops are discussed.     

Suitability of the tomato borer Tuta absoluta as prey for Macrolophus pygmaeus and Nesidiocoris tenuis

The tomato borer, Tuta absoluta (Meyrick) (Lep.: Gelechiidae), is an important tomato pest native to South America, which appeared in eastern Spain at the end of 2006. Prey suitability of T. absoluta eggs and larval instars was examined under laboratory conditions to evaluate whether two indigenous predators, Macrolophus pygmaeus (Rambur) and Nesidiocoris tenuis Reuter (Hem.: Miridae), can adapt to this invasive pest. Both predators preyed actively on T. absoluta eggs and all larval stages, although they preferred first‐instar larvae. Our results demonstrate that both mirids can adapt to this invasive pest, contributing to their value as biological control agents in tomato crops.     

Humic Fertilizer and Vermicompost Applied to the Soil Can Positively Affect Population Growth Parameters of <Emphasis Type="Italic">Trichogramma brassicae</Emphasis> (Hymenoptera: Trichogrammatidae) on Eggs of <Emphasis Type="Italic">Tuta absoluta</Emphasis> (Lepidoptera: Gelechiidae)

The tomato leaf miner, Tuta absoluta (Meyrick), is a devastating pest of tomato worldwide. One of the control measures of T. absoluta is the use of biological control agents, such as Trichogramma wasps. Interactions between natural enemies and insect pests may be affected by application of fertilizers, because changes in plant quality through the fertilizer application may therefore affect herbivore characteristics and suitability of them to parasitism. Laboratory tests were carried out to evaluate the life table parameters of Trichogramma brassicae Bezdenko on T. absoluta eggs reared on tomato plants treated either with vermicompost (40%), humic fertilizer (2 g/kg soil), or control (suitable mixture of field soil and sand). Population growth parameters of T. brassicae were affected by fertilizer treatments. Significant differences were found for immature life period and total fecundity of T. brassicae on the treatments. Differences of intrinsic rate of natural increase (r _m), finite rate of increase (λ), net reproductive rate (R ₀), mean generation time (T), and doubling time (DT) of T. brassicae among treatments were also significant. The lowest values of r _m, λ, and R ₀ were recorded for T. brassicae developed on T. absoluta eggs on control treatment, whereas the highest values of these parameters were observed on 2 g/kg humic fertilizer. Furthermore, T. brassicae had the shortest T and DT values on 2 g/kg humic fertilizer and 40% vermicompost treatments. Our results showed that application of humic fertilizer and vermicompost could positively affect population growth parameters of T. brassicae on eggs of T. absoluta fed on tomato plants.     

Assessment of Tuta absoluta yield loss in Iranian tomato crops

The South American tomato pinworm, Tuta absoluta Meyrick (Lepidoptera: Gelechiidae), a devastating pest of tomato plants, originates from South America but it has been considered an invasive pest for Iran since 2010. To assess crop loss caused by this pest, a study was conducted in a 600 m² field under a randomised complete block design. The impact of different initial densities of T. absoluta, including control (without infestation), one, two, four, and eight T. absoluta eggs on each host plant in the first and second year was investigated with an additional treatment (16 pinworm eggs per host plant) used in the second year. Yield attributes including length and width of fruits, numbers of infested and intact fruits along with their weights were sampled and assessed on a weekly basis. In addition, the correlation between the number of galleries and these characteristics was also investigated. Crop loss data demonstrated that the effect of different infestation levels on fruit size and the weight of intact fruits was not significant over both years. However, the effect of treatment on the percentage and weight of infested fruits was shown to be significantly different in 2014 and 2015. Furthermore, there was a negative significant correlation between the number of galleries and weight of infested fruits in both years. Collectively, these results will be useful for enhancing pest management programs of T. absoluta in Iran.     

Mutation in the ace‐1 gene of the tomato leaf miner (Tuta absoluta) associated with organophosphates resistance

The tomato leaf miner, Tuta absoluta (Lepidoptera: Gelechiidae), is a major invasive pest that has spread throughout many countries in the Mediterranean basin and parts of Asia over the last decade. The control of T. absoluta has relied heavily on the use of chemical insecticides, a strategy that has led to the evolution of resistance. In this study, biological and molecular methods were used to determine the susceptibility of five strains of T. absoluta to the organophosphate chlorpyrifos and to investigate the molecular mechanisms underlying resistance to this class of insecticides. High levels of resistance to chlorpyrifos were observed in all five strains tested. Cloning and sequencing of the gene encoding the organophosphate target site, ace‐1, of T. absoluta revealed the presence of an alanine to serine substitution at a position that has been previously linked with organophosphate resistance across a range of different insect and mite species. The presence of this mutation at high frequency in T. absoluta populations originating from various countries further supports the suggestion that the rapid expansion of this species is, in part, mediated by the resistance of this pest to chemical insecticides.     

A comparative life history study of two mirid bugs preying on Tuta absoluta and Ephestia kuehniella eggs on tomato crops: implications for biological control

The omnivorous predators Nesidiocoris tenuis (Reuter) and Macrolophus pygmaeus Rambur (Hemiptera: Miridae) are indigenous natural enemies that commonly inhabit tomato crops in the Mediterranean basin. Both predators are mass-reared and primarily released to control whiteflies, although recently they have also contributed to the control of the invasive tomato pest Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae). The life history traits of these two predators have been studied in the laboratory under the conditions of being fed exclusively the eggs of T. absoluta or the eggs of the factitious prey Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae). Immature stages of both predator species successfully developed while preying on eggs of T. absoluta. However, the mature M. pygmaeus females produced significantly lower numbers of offspring in comparison to the offspring produced when preying on E. kuehniella eggs. This resulted in higher than expected demographic indexes for N. tenuis when compared to M. pygmaeus (e.g., the intrinsic rates of increase were 0.127 and 0.005, respectively). Our results support previous studies on the potential of N. tenuis has as biological control agent of T. absoluta, and indicate that the role of M. pygmaeus in controlling T. absoluta in the absence of other food sources is possibly limited.     

Adaptation of indigenous larval parasitoids to Tuta absoluta (Lepidoptera: Gelechiidae) in Italy

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a serious threat to tomato (Solanum lycopersicum L.) crops in South America. In Europe, after its first detection in Spain in 2006, it rapidly spread through the Mediterranean basin, reaching Italy 2 yr later. The aim of our work was to find indigenous effective biological control agents and to evaluate their potential role in the control of larval populations of T. absoluta in controlled conditions. Nine species of larval parasitoids emerged from field-collected tomato leaves infested by T. absoluta. The most abundant, Necremnus near artynes (Walker) and Necremnus near tidius (Walker) (Hymenoptera: Eulophidae), were tested in laboratory parasitism trials. Furthermore, because the species N. artynes and N. tidius are each reported in literature as an ectoparasitoid of Cosmopterix pulchrimella Chambers (Lepidoptera: Cosmopterigidae) on upright pellitory plants, olfactometer bioassays were performed to assess the response of our parasitoids to the odors of tomato and pellitory leaves infested by T absoluta and C. pulchrimella, respectively, compared with healthy ones. Both Necremnus species showed good adaptation to the invasive pest, and we observed a high larval mortality of T. absoluta because of host feeding and parasitism. Even olfactory responses highlighted a preference of both wasps for tomato plants infested by the exotic pest. These preliminary results demonstrated a high suitability of these indigenous natural enemies for controlling T. absoluta. Further investigations are needed to confirm their role as potential biological agents in commercial tomato plantations.     

Are egg parasitoids of the genus Trichogramma (Hymenoptera: Trichogrammatidae) promising biological control agents for regulating the invasive Box tree pyralid,Cydalima perspectalis (Lepidoptera: Crambidae)?

The Box tree pyralid, Cydalima perspectalis Walker (Lepidoptera: Crambidae), is an invasive species in Europe, not yet regulated by natural enemies. It is highly destructive to ornamental and native Buxus due to defoliation by larval feeding. The ability of eight Trichogramma species (T. brassicae Bezdenko, T. bourarachae Pintureau & Babault, T. cacoeciae Marchal, T. cordubensis Vargas & Cabello, T. dendrolimi Matsumura, T. evanescens Westwood, T. nerudai Pintureau & Gerding and T. pintoi Voegele) to parasitise and locate eggs of C. perspectalis was assessed in the laboratory. In direct observation tests, all species started to examine and oviposit into exposed eggs. Resulting egg parasitism was low and ranged between 4% and 20% for most of the parasitoid species after three days of exposure. Only T. dendrolimi parasitised significantly more host eggs with parasitism rates above 40%. On average, two to three progeny emerged from one host egg and the resulting sex ratio was female-biased, indicating suitability of the target host for progeny production. In subsequent cage tests on potted Buxus plants, again T. dendrolimi was the most efficient species in locating host eggs on foliage. We conclude that C. perspectalis eggs are attractive to various Trichogramma species. T. pintoi had the highest number of egg encounters and active females. T. dendrolimi females were most able to locate host eggs and to oviposit and produce successfully on them. Nonetheless, further study is needed to clarify the suspected role of potentially egg defensive mechanisms that may impede higher parasitisation in C. perspectalis.     

Natural occurrence of Wolbachia-infected and uninfected Trichogramma species in tomato fields in Portugal

Minute egg parasitoids of the genus Trichogramma (Hymenoptera; Trichogrammatidae) are promising candidates for biological control of lepidopteran pests in tomato in Portugal. This certainly applies to native Trichogramma strains that have thelytokous reproduction, i.e., produce only daughters. In Trichogramma wasps, thelytoky is mostly induced by the intracellular bacterium Wolbachia. In this study, we carried out a field survey of native Trichogramma species in four locations in Ribatejo, the main processing tomato region of Portugal, and determined the prevalence of Wolbachia in those species. Five Trichogramma species were found to emerge from lepidopteran eggs collected in the field, namely Trichogramma bourarache, Trichogramma cordubensis, Trichogramma evanescens, Trichogramma pintoi, and Trichogramma turkestanica. T. evanescens and T. pintoi were by far the dominating species representing, respectively, 64.9 and 26.4% of the trichogrammatids collected. Total natural parasitism rates of the collected lepidopteran eggs by Trichogramma wasps ranged from 28.2 to 64.6%. Three Trichogramma species were found to be infected with Wolbachia, namely T. cordubensis, T. evanescens, and T. turkestanica. All the wasp broods belonging to T. cordubensis were infected, whereas low infection rates were found in T. evanescens (0.9% of the broods) and T. turkestanica (4.5% of the broods). The latter represents the first record of a Wolbachia infection in T. turkestanica. Sequencing of the Wolbachia surface protein, wsp, revealed this Wolbachia infection to be related to other Wolbachia infections in Trichogramma wasps. As Wolbachia-infected thelytokous strains exist for T. evanescens, the most abundant Trichogramma species naturally occurring in the tomato fields of the Ribatejo region, this species offers interesting and powerful options for biological control of lepidopteran pests in processing tomato in this region.