Salvatore Arpaia

Biological invasion of European tomato crops by Tuta absoluta: ecology, geographic expansion and prospects for biological control.

The tomato leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating pest of tomato originating from South America. After its initial detection in eastern Spain in 2006, it rapidly invaded various other European countries and spread throughout the Mediterranean basin. If no control measures are taken, then the pest can cause up to 80–100% yield losses in tomato crops in recently invaded areas and may pose a threat to both greenhouse and open-field tomato production. The exceptional speed and extent of T. absoluta invasion have called for studies documenting its biology and ecology, while indicating an urgent need for efficient and sustainable management methods. The development of approaches to manage T. absoluta would be facilitated through a detailed revision of information on this pest in its area of origin. This review combines information on the invasion by T. absoluta, its ecology, and potential management strategies, including data that may help the implementation of efficient biological control programs. These programs, together with a variety of other management tactics, may allow efficient integrated pest management of T. absoluta in Europe and Mediterranean Basin countries.

The impact of transgenic plants on natural enemies: a critical review of laboratory studies

We reviewed laboratory tests which studied the impact of genetically modified plants on arthropod natural enemies. A total of 18 species of predators and 14 species of parasitoids have been tested, most in only a few experiments. Certain groups (braconid wasps) or species (the green lacewing, Chrysoperla carnea) have attracted much effort, while representatives of others, including whole orders (e.g., Diptera), have never had a species tested. We conclude that laboratory tests are not the ‘worst case’ scenarios intended by the experimental designs, and are not often ecologically realistic: they typically provided ad libitum feeding, no prey choice, single prey type, no combination of stress factors and usually uniform temperatures. None of these are representative of field conditions, yet most could be easily mimicked in more complex laboratory tests. In most cases (94.6%), the studies were unable to indicate the level of power required to detect any impact. Small sample size and large variability are factors that mask all but very large differences in potential effects. For predators, 126 parameters were quantified, most commonly including survival/mortality (37 cases), development time (22), and body mass/size (20). For parasitoids, 128 parameters were quantified, the majority involving lectins or proteinase inhibitors. Most frequent measurements were: fecundity (23 experiments), adult longevity, extent of parasitism (17 each), body size, mortality, and larval development time. An aggregative scoring (summarising all quantified parameters) indicated that the laboratory tests quantified a remarkable number of cases (30% for predators, 39.8% for parasitoids), where the impacts of the genetically modified plant were significantly negative. These involve various parameters, organisms, test methods, and significance levels, but collectively they indicate that the use of genetically modified crops may result in negative effects on the natural enemies of crop pests.

Transgenic Insecticidal Crops and Natural Enemies: A Detailed Review of Laboratory Studies

ABSTRACT This review uses a data-driven, quantitative method to summarize the published, peer-reviewed literature about the impact of genetically modified (GM) plants on arthropod natural enemies in laboratory experiments. The method is similar to meta-analysis, and, in contrast to a simple author-vote counting method used by several earlier reviews, gives an objective, data-driven summary of existing knowledge about these effects. Significantly more non-neutral responses were observed than expected at random in 75% of the comparisons of natural enemy groups and response classes. These observations indicate that Cry toxins and proteinase inhibitors often have non-neutral effects on natural enemies. This synthesis identifies a continued bias toward studies on a few predator species, especially the green lacewing, Chrysoperla cornea Stephens, which may be more sensitive to GM insecticidal plants (16.8% of the quantified parameter responses were significantly negative) than predators in general (10.9% significantly negative effects without C. cornea). Parasitoids were more susceptible than predators to the effects of both Cry toxins and proteinase inhibitors, with fewer positive effects (18.0%, significant and nonsignificant positive effects combined) than negative ones (66.1%, significant and nonsignificant negative effects combined). GM plants can have a positive effect on natural enemies (4.8% of responses were significantly positive), although significant negative (21.2%) effects were more common. Although there are data on 48 natural enemy species, the database is still far from adequate to predict the effect of a Bt toxin or proteinase inhibitor on natural enemies.

A mathematical model of exposure of non- target Lepidoptera to Bt-maize pollen expressing Cry1Ab within Europe

Genetically modified (GM) maize MON810 expresses a Cry1Ab insecticidal protein, derived from Bacillus thuringiensis (Bt), toxic to lepidopteran target pests such as Ostrinia nubilalis. An environmental risk to non-target Lepidoptera from this GM crop is exposure to harmful amounts of Bt-containing pollen deposited on host plants in or near MON810 fields. An 11-parameter mathematical model analysed exposure of larvae of three non-target species: the butterflies Inachis io (L.), Vanessa atalanta (L.) and moth Plutella xylostella (L.), in 11 representative maize cultivation regions in four European countries. A mortality–dose relationship was integrated with a dose–distance relationship to estimate mortality both within the maize MON810 crop and within the field margin at varying distances from the crop edge. Mortality estimates were adjusted to allow for physical effects; the lack of temporal coincidence between the susceptible larval stage concerned and the period over which maize MON810 pollen is shed; and seven further parameters concerned with maize agronomy and host-plant ecology. Sublethal effects were estimated and allowance made for aggregated pollen deposition. Estimated environmental impact was low: in all regions, the calculated mortality rate for worst-case scenarios was less than one individual in every 1572 for the butterflies and one in 392 for the moth.

Estimating the effects of Cry1F Bt‐maize pollen on non‐target Lepidoptera using a mathematical model of exposure

Summary 1. In farmland biodiversity, a potential risk to the larvae of non‐target Lepidoptera from genetically modified (GM) Bt‐maize expressing insecticidal Cry1 proteins is the ingestion of harmful amounts of pollen deposited on their host plants. A previous mathematical model of exposure quantified this risk for Cry1Ab protein. We extend this model to quantify the risk for sensitive species exposed to pollen containing Cry1F protein from maize event 1507 and to provide recommendations for management to mitigate this risk. 2. A 14‐parameter mathematical model integrating small‐ and large‐scale exposure was used to estimate the larval mortality of hypothetical species with a range of sensitivities, and under a range of simulated mitigation measures consisting of non‐Bt maize strips of different widths placed around the field edge. 3. The greatest source of variability in estimated mortality was species sensitivity. Before allowance for effects of large‐scale exposure, with moderate within‐crop host‐plant density and with no mitigation, estimated mortality locally was <10% for species of average sensitivity. For the worst‐case extreme sensitivity considered, estimated mortality locally was 99·6% with no mitigation, although this estimate was reduced to below 40% with mitigation of 24‐m‐wide strips of non‐Bt maize. For highly sensitive species, a 12‐m‐wide strip reduced estimated local mortality under 1·5%, when within‐crop host‐plant density was zero. Allowance for large‐scale exposure effects would reduce these estimates of local mortality by a highly variable amount, but typically of the order of 50‐fold. 4. Mitigation efficacy depended critically on assumed within‐crop host‐plant density; if this could be assumed negligible, then the estimated effect of mitigation would reduce local mortality below 1% even for very highly sensitive species. 5.  Synthesis and applications. Mitigation measures of risks of Bt‐maize to sensitive larvae of non‐target lepidopteran species can be effective, but depend on host‐plant densities which are in turn affected by weed‐management regimes. We discuss the relevance for management of maize events where cry1F is combined (stacked) with a herbicide‐tolerance trait. This exemplifies how interactions between biota may occur when different traits are stacked irrespective of interactions between the proteins themselves and highlights the importance of accounting for crop management in the assessment of the ecological impact of GM plants.

The usefulness of a mathematical model of exposure for environmental risk assessment.

We respond to the Comment of Lang et al . [[1][1]] regarding our mathematical model [[2][2]] of exposure of non-target Lepidoptera to Bt -maize pollen expressing Cry1Ab within Europe. Lang et al . remark on the degree to which the model was subject to uncertainty. Perry et al . [[2][2]] did indeed

Shared flowering phenology, insect pests, and pathogens among wild, weedy, and cultivated rice in the Mekong Delta, Vietnam: implications for transgenic rice

Many varieties of transgenic rice are under development in countries where wild and weedy relatives co-occur with the crop. To evaluate possible risks associated with pollen-mediated transgene dispersal, we conducted a two-year survey in Vietnam to examine overlapping flowering periods of rice (Oryza sativa L.), weedy rice (O. sativa), and wild rice (O. rufipogon Griff.), all of which are inter-fertile. We surveyed populations in two regions of the Mekong Delta, northern and southern, and at three sites in each of three habitats per region: fresh water, saline water, and acid sulfate soil. Weedy rice frequently flowered simultaneously with neighboring cultivated rice plants. Flowering was more seasonal in wild rice and often peaked in November and December. Peak flowering times of wild rice overlapped with adjacent rice fields at all of the saline sites and half of the acid sulfate sites. The longer flowering season of wild rice ensured that crop-to-wild gene flow was possible in fresh water habitats as well. Our second objective was to determine whether wild and weedy rice populations are exposed to pests that could be targeted by future transgenes, which may then provide fitness benefits. These populations shared many pathogen and insect herbivore species with cultivated rice (leaffolder, locust, cricket, planthoppers, rice bug, stem borer, sheath blight, blast, bacterial leaf blight, and brown spot). Damage by leaffolders and locusts was the most frequently observed insect feeding damage on all three rice types. Indicator species analysis revealed that most of the insect herbivores were associated with particular habitats, demonstrating the importance of broad geographic sampling for transgenic rice risk assessment. These survey data and the strong likelihood of gene flow from cultivated rice suggest that further studies are needed to examine the effects of transgenic traits such as resistance to pests on the abundance of wild and weedy rice.

Feeding behaviour and reproductive biology of Colorado potato beetle adults fed transgenic potatoes expressing the Bacillus thuringiensis Cry3B endotoxin

Colorado potato beetle (Leptinotarsa decemlineata Say) adult longevity and fecundity were studied on transgenic potato clones expressing a Cry3B endotoxin of Bacillus thuringiensis (Bt). Adult longevity and fitness were studied for the first 3 weeks after emergence. Beetle reproductive biology on highly resistant clones, intermediary resistant clones and control potato plants was monitored by dissecting females after 7–15 days of feeding and also by analysing haemolymph protein content after 3 days of feeding. Feeding behaviour on transgenic plants expressing high toxin concentrations and on control plants was monitored individually for 36 newly emerged adult beetles feeding on leaf disks during the first two meals. Lethal Time50 for adult beetles feeding on transgenic clones as the sole source of food was not significantly shorter than for beetles on control clones reared in a growth chamber. Differences tended to be larger when the experiment was conducted in a greenhouse with a less optimal temperature range (LT50= 9.52 and 10.45 days for two transgenic clones and 13.86 for control). In contrast, female egg production on transgenic plants was almost totally inhibited. Dissection studies indicated that adult males living on high‐level Bt‐expressing transgenic potatoes were still able to mate and produce mobile sperm, but the females were impaired in their reproductive ability since their ovaries were generally not fully developed. An examination of the haemolymph revealed the protein concentration in females living on transgenic plants to be dramatically reduced (≅ 50%), and electrophoresis showed a reduced content of vitellogenin in these samples.

Interactions between Bt-expressing tomato and non-target insects: the aphid Macrosiphum euphorbiae and its natural enemies.

Bacillus thuringiensis Berliner toxins are highly specific in terms of insect pest targeted. For example, Cry3Bb is active against Coleoptera. As a consequence, in the framework of using genetically modified plants expressing different Bt toxins in sustainable agriculture, there is a growing interest about the impact of these crops on the environment and on non-target organisms. The effects of genetically modified Bt-tomato plants expressing the toxin Cry3Bb on the aphid Macrosiphum euphorbiae (Thomas) and its natural enemies, the generalist predator Macrolophus caliginosus Wagner and the endoparasitoid wasp Aphidius ervi Haliday, were assessed under laboratory conditions. No significant differences between performance of M. euphorbiae were observed on genetically modified tomato plants (line UC82Bt) with respect to their near-isogenic control line (line UC82). Immunoassays did not detect Cry3Bb protein in M. euphorbiae developing on Bt-expressing tomato plants. Similarly, no significant differences were found on the longevity and prey consumption of M. caliginosus when fed aphids reared on UC82Bt or on UC82. Moreover, the genetic modification did not affect the attractiveness of uninfested tomato plants toward A. ervi. It is therefore concluded that Cry3Bb-expressing tomato plants did not show any acute adverse effects on the biological parameters of the non-target herbivore M. euphorbiae or its natural enemies, M. caliginosus and A. ervi, under laboratory conditions.

Effects of azadirachtin after systemic uptake into Brassica oleracea on larvae of Pieris brassicae

Petioles of intact cabbage leaves were placed in azadirachtin solutions for 24 h. Azadirachtin or active metabolites of this compound appeared to be translocated systemically as performance of first instar larvae of Pieris brassicae L. placed on disks from those leaves was affected. The position within a leaf from which disks were excised did not affect the extent of these effects, indicating a homogeneous distribution of active compounds in the laminar tissue. At doses ranging from 1–10 ppm of azadirachtin in the solution taken up by the leaves, gain of larval fresh weight and leaf area consumed were reduced. These effects were dose dependent up to 60–90 ppm. At 10 ppm and higher doses no larva reached the second instar within 72 h. Mortality during the 72 h period was highest at 10 ppm, due to the coincidence of lethal action with ecdysis in combination with the effect on developmental rate. Azadirachtin solutions applied directly to the surface of leaf disks in order to produce known doses on a leaf fresh weight basis gave results similar to the systemic treatment for weight gain, leaf area consumed and developmental rate, while mortality was higher in the latter treatment.