Characterising insect plant host relationships facilitates understanding multiple host use

Weed biocontrol relies on host specificity testing, usually carried out under quarantine conditions to predict the future host range of candidate control agents. The predictive power of host testing can be scrutinised directly with Aconophora compressa, previously released against the weed Lantana camara L. (lantana) because its ecology in its new range (Australia) is known and includes the unanticipated use of several host species. Glasshouse based predictions of field host use from experiments designed a posteriori can therefore be compared against known field host use. Adult survival, reproductive output and egg maturation were quantified. Adult survival did not differ statistically across the four verbenaceous hosts used in Australia. Oviposition was significantly highest on fiddlewood (Citharexylum spinosum L.), followed by lantana, on which oviposition was significantly higher than on two varieties of Duranta erecta (‘‘geisha girl’’ and ‘‘Sheena’s gold’’; all Verbenaceae). Oviposition rates across Duranta varieties were not significantly different from each other but were significantly higher than on the two non-verbenaceous hosts (Jacaranda mimosifolia D. Don: Bignoneaceae (jacaranda) and Myoporum acuminatum R. Br.: Myoporaceae (Myoporum)). Production of adult A. compressa was modelled across the hosts tested. The only major discrepancy between model output and their relative abundance across hosts in the field was that densities on lantana in the field were much lower than predicted by the model. The adults may, therefore, not locate lantana under field conditions and/or adults may find lantana but leave after laying relatively few eggs. Fiddlewood is the only primary host plant of A. compressa in Australia, whereas lantana and the others are used secondarily or incidentally. The distinction between primary, secondary and incidental hosts of a herbivore species helps to predict the intensity and regularity of host use by that herbivore. Populations of the primary host plants of a released biological control agent are most likely to be consistently impacted by the herbivore, whereas secondary and incidental host plant species are unlikely to be impacted consistently. As a consequence, potential biocontrol agents should be released only against hosts to which they have been shown to be primarily adapted.

Classical Biological Control for the Protection of Natural Ecosystems.

Of the 70 cases of classical biological control for the protection of nature found in our review, there were fewer projects against insect targets (21) than against invasive plants (49), in part, because many insect biological control projects were carried out against agricultural pests, while nearly all projects against plants targeted invasive plants in natural ecosystems. Of 21 insect projects, 81% (17) provided benefits to protection of biodiversity, while 48% (10) protected products harvested from natural systems, and 5% (1) preserved ecosystem services, with many projects contributing to more than one goal. In contrast, of the 49 projects against invasive plants, 98% (48) provided benefits to protection of biodiversity, while 47% (23) protected products, and 25% (12) preserved ecosystem services, again with many projects contributing to several goals. We classified projects into complete control (pest generally no longer important), partial control (control in some areas but not others), and "in progress," for projects in development for which outcomes do not yet exist. For insects, of the 21 projects discussed, 59% (13) achieved complete control of the target pest, 18% (4) provided partial control, and 41% (9) are still in progress. By comparison, of the 49 invasive plant projects considered, 27% (13) achieved complete control, while 33% (16) provided partial control, and 47% (24) are still in progress. For both categories of pests, some projects' success ratings were scored twice when results varied by region. We found approximately twice as many projects directed against invasive plants than insects and that protection of biodiversity was the most frequent benefit of both insect and plant projects. Ecosystem service protection was provided in the fewest cases by either insect or plant biological control agents, but was more likely to be provided by projects directed against invasive plants, likely because of the strong effects plants exert on landscapes. Rates of complete success appeared to be higher for insect than plant targets (59% vs 27%), perhaps because most often herbivores gradually weaken, rather than outright kill, their hosts, which is not the case for natural enemies directed against pest insects. For both insect and plant biological control, nearly half of all projects reviewed were listed as currently in progress, suggesting that the use of biological control for the protection of wildlands is currently very active.

Relative host plant species use by the lantana biological control agent Aconophora compressa (Membracidae) across its native and introduced ranges

Aconophora compressa Walker (Hemiptera: Membracidae) was released in 1995 against the weed lantana in Australia, and is now found on multiple host plant species. The intensity and regularity at which A. compressa uses different host species was quantified in its introduced Australian range and also its native Mexican range. In Australia, host plants fell into three statistically defined categories, as indicated by the relative rates and intensities at which they were used in the field. Fiddlewood (Citharexylum spinosum L.: Verbenaceae) was used much more regularly and at higher densities than any other host sampled, and alone made up the first group. The second group, lantana (Lantana camara L.: Verbenaceae; pink variety) and geisha girl (Duranta erecta L.: Verbenaceae), were used less regularly and at much lower densities than fiddlewood. The third group, Sheena’s gold (another variety of D. erecta), jacaranda (Jacaranda mimosifolia D. Don: Bignoniaceae) and myoporum (Myoporum acuminatum R. Br.: Myoporaceae), were used infrequently and at even lower densities. In Mexico, the insect was found at relatively low densities on all hosts relative to those in Australia. Densities were highest on L. urticifolia, D. erecta and Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae), which were used at similar rates to one another. It was found also on a few other verbenaceous and non-verbenaceous host species but at even lower densities. The relative rate at which Citharexylum spp. and L. urticifolia were used could not be assessed in Mexico because A. compressa was found on only one plant of each species in areas where these host species co-occurred. The low rate at which A. compressa occurred on fiddlewood in Mexico is likely to be an artefact of the short-term nature of the surveys or differences in the suites of Citharexylum and Lantana species available there. These results provide further incentive to insist on structured and quantified surveys of non-target host use in the native range of potential biological control agents prior to host testing studies in quarantine.

Assessing the biological control potential of an adventitiously-established "pest", Scirtothrips aurantii (Faure), on a weed, Bryophyllum delagoense (Eckl. & Zeyh.), in Queensland

South African citrus thrips (Scirtothrips aurantii) established adventitiously in Australia. Although it is a major horticultural pest in Africa, it is now advocated as a possible biological control agent against Bryophyllum delagoense Eckl. & Zeyh. (Crassulaceae). To evaluate the biocontrol potential of S. aurantii a two year field study was conducted on the western Darling Downs of southern Queensland. Imidacloprid insecticide was applied to two quadrats at each of 18 field sites to assess, in the absence of S. aurantii, the persistence of individual plants and to quantify propagule production and recruitment by this declared weed. A third quadrat was left, as a control, to be infested naturally by S. aurantii. When released from herbivory by thrips in the field, plants grew significantly more, flowered more, and were significantly more fecund than plants in the quadrats with S. aurantii. Increases in growth and fecundity translated into significantly increased plant numbers but not increased recruitment. Recruitment even declined in experimental quadrats, through the indirect effects of releasing plants from herbivory. Field sampling also revealed that S. aurantii may be sensitive to seasonal climatic fluctuations. These and other local climatic influences may limit the biological control potential of the insect.

Biological Control of Eucalypt Pests Overseas and Australia

This project aims to address the growing need for a coordinated approach to research into the biological control of Australian eucalypt insect pests by scoping the formation of a Centre in Australia which would (a) coordinate the evaluation and provision of biological control agents (initially to South Africa and Brazil, but in future years more widely), (b) research the role natural enemies play in pest population regulation in Australian eucalypt plantations and how this may be enhanced as a management tool, and (c) form a network focussed on forest biosecurity with an emphasis on eucalypt pests and pathogens.

CRC50089 Grain Insect Ecology

Resistance to phosphine in target pests threatens market access for Australian grain. While the grains industry is now attempting to develop an effective and sustainable strategy to manage this resistance, action is severely limited by significant gaps in our knowledge of the key ecological factors that influence the development of resistance. There is a need to research this information as a foundation for a rational approach to managing phosphine resistance in the Australian grains industry. Research outcomes: The project has provided critical research methodologies and preliminary data to fill the large gaps in our knowledge of the ecology of two key pests, Rhyzopertha dominica and Tribolium castaneum, and how this may drive the development of phosphine resistance. This information will contribute to the groundwork for future research needed to provide a scientific basis for a rational resistance management strategy.

Cat's claw creeper leaf-mining jewel beetle Hylaeogena jureceki Obenberger (Coleoptera: Buprestidae), a host-specific biological control agent for Dolichandra unguis-cati (Bignoniaceae) in Australia

Cat's claw creeper, Dolichandra unguis-cati (L.) L.G. Lohman (syn: Macfadyena unguis-cati (L.) A.H. Gentry) (Bignoniaceae), a major environmental weed in Queensland and New South Wales, is a Weed of National Significance and an approved target for biological control. A leaf-mining jewel beetle, Hylaeogena jureceki Obenberger (Coleoptera: Buprestidae), first collected in 2002 from D. unguis-cati in Brazil and Argentina, was imported from South Africa into a quarantine facility in Brisbane in 2009 for host-specificity testing. H. jureceki adults chew holes in leaves and lay eggs on leaf margins and the emerging larvae mine within the leaves of D. unguis-cati. The generation time (egg to adult) of H. jureceki under quarantine conditions was 55.4 ± 0.2 days. Host-specificity trials conducted in Australia on 38 plant species from 11 families supplement and support South African studies which indicated that H. jureceki is highly host-specific and does not pose a risk to any non-target plant species in Australia. In no-choice tests, adults survived significantly longer (>32 weeks) on D. unguis-cati than on non-target test plant species (<3 weeks). Oviposition occurred on D. unguis-cati and one non-target test plant species, Citharexylum spinosum (Verbenaceae), but no larval development occurred on the latter species. In choice tests involving D. unguis-cati, C. spinosum and Avicennia marina (Avicenniaceae), feeding and oviposition were evident only on D. unguis-cati. The insect was approved for field release in Australia in May 2012.

Impact of a biological control agent, Chiasmia assimilis, on prickly acacia (Acacia nilotica ssp. indica) seedlings

A leaf-feeding geometrid, Chiasmia assimilis (Warren), was introduced into northern Queensland from South Africa in 2002 as a biological control agent for the invasive woody weed, prickly acacia, Acacia nilotica subsp. indica (Bentham) Brenan. The insect established in infestations in coastal areas between the townships of Ayr and Bowen where the larvae periodically cause extensive defoliation at some localities during summer and autumn. The impact of this herbivory on a number of plant parameters, including shoot length, basal stem diameter, root length, number of leaves, number of branches, and above and below ground biomass was investigated at one coastal site through an insect exclusion trial using potted seedlings and regular spray applications of a systemic insecticide to exclude the biological control agent. Half the seedlings, both sprayed and unsprayed, were placed beneath the prickly acacia canopy, the other half were placed in full sunlight. Larvae of C. assimilis were found on unsprayed seedlings in both situations. The effects of herbivory, however, were significant only for seedlings grown beneath the canopy. At the end of the five-month trial period, shoot length of these seedlings was reduced by 30%, basal stem diameter by 44%, root length by 15%, number of leaves by 97%, above ground biomass by 87%, and below ground biomass by 77% when compared to sprayed seedlings. Implications are that the insect, where established, may reduce seedling growth beneath existing canopies and in turn may help limit the formation of dense infestations. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.

Biological Control of Oilseed Rape Pests with Entomopathogenic Nematodes

Biological control techniques attract increasing attention as one of the sustainable alternatives to pesticide use in integrated pest management programs. In order to develop sustainable pest management methods for arable crops based on entomopathogenic nematodes (EPN), their efficacy and persistence needed to be investigated, and an economically feasible delivery system had to be developed. In this study, first a survey of entomopathogens was conducted, and a system approach was tested, using the oilseed Brassica (OSB) growing system (OSB, spring wheat, and red clover) as a model. The system approach aimed at determining the potential of Steinernema feltiae (Filipjev) for the control of OSB pests, developing OSB rotation schemes that support EPN persistence, and investigating the impact of the selected biotic and abiotic factors on efficacy and persistence of EPN. This study employed abductive logic (which employs constant interplay between the theory and empirical observation), quantitative methods, and a case study on OSB. Laboratory and field experiments were carried out, and two types of pathogen surveys. A horizontal survey included OSB fields across Estonia, Germany, Poland, Sweden and the UK, while a vertical survey included sampling from two sets of differently managed experimental fields during three years. A new approach was introduced for measuring occurrence, where the prevalence and relative intensity of entomopathogens, biotic agents, and unidentified insect antagonists were determined. The effect of dose, timing, and the application method on S. feltiae in the control of pests in OSB, and the potential of a controlled release delivery system (CRS) were evaluated in the field. Studies on the impact of selected biotic and abiotc factors (Brassica plant, bait insects, developmental stages of Meligethes aeneus Fab., Isaria fumosorosea Wize (Ifr), and organic and synthetic fertilizers) on the efficacy of S. feltiae were conducted in the laboratory. Persistence of S. feltiae in the OSB growing system, and the effect of dose, timing, and the application method, was assessed in the field as part of the efficacy experiments. The impact of selected biotic and abiotic factors on S. feltiae persistence was assessed in laboratory experiments. The pathogen survey showed that the occurrence of entomopathogens is low in the OSB growing system, and that a management system causing less disturbance (ICM) to the soil increases the relative intensity of insect parasitic nematodes and other insect antagonists. A longer study period is required to show any possible impact of ICM on the relative intensity of entomopathogenic fungi, or on the prevalence of entomopathogens. Two different measures of the occurrence yielded different results: the relative intensity revealed the difference between the two different crop management methods, while prevalence did not. The highest efficacy of S. feltiae was achieved by using a low dose and targeting all stages of M. aeneus. When only the larval stage was targeted, the application method and dose had no significant effect. The CRS decreased the pest abundance significantly more than the surface application method. S. feltiae persisted in the OSB fields in Finland for several months, but did not survive the winter. The strain survived for 7 months when it was applied in autumn in Germany, but its populations declined rapidly after winter. The examined biotic and abiotic factors had variable impacts on S. feltiae efficacy and persistence. The two measures, prevalence and relative intensity of entomopathogens, gave valuable information for their use in biocontrol programs. The recommended biocontrol strategy for OSB growing in Finland is inundation and seasonal inoculation of EPN. The impact of some biotic and abiotic factors on S. feltiae efficacy and persistence is significant, and can be used to improve the efficacy of EPN. The CRS is a novel alternative for EPN application, and should also be considered for use on other crops. Keywords: Biological control, inundation, inoculation, conservation, formulation, slow release method, crop rotation, Entomopathogenic nematodes, Steinernema feltiae, oilseed rape pests, Meligethes aeneus, Phyllotreta spp., occurrence, prevalence, intensity, efficacy, persistence, field, Isaria fumosorosea, biotic factors, abiotic factors, interaction, impact, insect stages, integrated crop management, standard (conventional) crop management

Field Assessments of Control Agents for Lesser Mealworm (Coleoptera: Tenebrionidae) Using Litter Sampling

Spinosad, diatomaceous earth, and cyfluthrin were assessed on two broiler farms at Gleneagle and Gatton in southeastern Queensland, Australia in 2004-2005 and 2007-2009, respectively to determine their effectiveness in controlling lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Insecticide treatments were applied mostly to earth or 'hard' cement floors of broiler houses before the placement of new bedding. Efficacy of each agent was assessed by regular sampling of litter and counting of immature stages and adult beetles, and comparing insect counts in treatments to counts in untreated houses. Generally, the lowest numbers of lesser mealworm were recorded in the house with hard floors, these numbers equalling the most effective spinosad applications. The most effective treatment was a strategic application of spinosad under feed supply lines on a hard floor. In compacted earth floor houses, mean numbers of lesser mealworms for two under-feed-line spinosad treatments (i.e., 2-m-wide application at 0.18 g of active insecticide (g [AI]) in 100-ml water/m(2), and 1-m-wide application at 0.11 g ([AI] in 33-ml water/m(2)), and an entire floor spinosad treatment (0.07 g [AI] in 86-ml water/m2) were significantly lower (i.e., better control) than those numbers for cyfluthrin, and no treatment (controls). The 1-m-wide under-feed-line treatment was the most cost-effective dose, providing similar control to the other two most effective spinosad treatments, but using less than half the active component per broiler house. No efficacy was demonstrated when spinosad was applied to the surface of bedding in relatively large volumes of water. All applications of diatomaceous earth, applied with and without spinosad, and cyfluthrin at the label rate of 0.02 g (AI)/100-ml water/m(2) showed no effect, with insect counts not significantly different to untreated controls. Overall, the results of this field assessment indicate that cyfluthrin (the Australian industry standard) and diatomaceous earth were ineffective on these two farms and that spinosad can be a viable alternative for broiler house use.

Mitochondrial DNA diversity of Cleruchoides noackae (Hymenoptera: Mymaridae): a potential biological control agent for Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae)

Carpintero and Dellap, (Hemiptera: Thaumastocoridae) is a native Australian sap-feeding insect that has become invasive and seriously damaging to commercially grown in the Southern Hemisphere. Lin and Huber (Hymenoptera: Mymaridae) was recently discovered as an egg parasitoid of the Thaumastocoridae in Australia. Mitochondrial DNA (mtDNA; cytochrome oxidase subunit I, COI) sequence diversity amongst 104 individuals from these native populations revealed 24 sequence haplotypes. The COI haplotypes of individuals collected from the Sydney and Southeast Queensland clustered in distinct groups, indicating limited spread of the insect between the regions. Individuals collected from Perth in Western Australia were represented by four COI haplotypes. Although this population is geographically more isolated from other populations, two COI haplotypes were identical to haplotypes found in the Sydney region. The results suggest that has recently been introduced into Perth, possibly from the Sydney area. The high mtDNA diversity and limited spread that is suggested for is in contrast to the lack of geographic associated mtDNA diversity and extensive spread of . If implemented as a biological control agent, this factor will need to be considered in collecting and releasing .

Biological control of prickly acacia: Current research and future prospects

Prickly acacia, Vachellia nilotica subsp. indica (syn. Acacia nilotica subsp. indica) (Fabaceae), a major weed in the natural grasslands of western Queensland, has been a target of biological control since the 1980s with limited success to date. Surveys in India, based on genetic and climate matching, identified five insects and two rust pathogens as potential agents. Host-specificity tests were conducted for the insects in India and under quarantine conditions in Australia, and for the rust pathogens under quarantine conditions at CABI in the UK. In no-choice tests, the brown leaf-webber, Phycita sp. A, (Lepidoptera: Pyralidae) completed development on 17 non-target plant species. Though the moth showed a clear preference for prickly acacia in oviposition choice trials screening of additional test-plant species was terminated in view of the potential non-target risk. The scale insect Anomalococcus indicus (Hemiptera: Lecanodiaspididae) developed into mature gravid females on 13 out of 58 non-target plant species tested. In the majority of cases very few female scales matured but development was comparable to that on prickly acacia on four of the non-target species. In multiple choice tests, the scale insect showed a significant preference for the target weed over non-target species tested. In a paired-choice trial under field conditions in India, crawler establishment occurred only on prickly acacia and not on the non-target species tested. Further choice trials are to be conducted under natural field conditions in India. A colony of the green leaf-webber Phycita sp. B has been established in quarantine facilities in Australia and host-specificity testing has commenced. The gall-rust Ravenelia acaciae-arabicae and the leaf-rust Ravenelia evansii (Puccineales: Raveneliaceae) both infected and produced viable urediniospores on Vachellia sutherlandii (Fabaceae), a non-target Australian native plant species. Hence, no further testing with the two rust species was pursued. Inoculation trials using the gall mite Aceria liopeltus (Acari: Eriophyidae) from V. nilotica subsp. kraussiana in South Africa resulted in no gall induction on V. nilotica subsp. indica. Future research will focus on the leaf-weevil Dereodus denticollis (Coleoptera: Curculionidae) and the leaf-beetle Pachnephorus sp. (Coleoptera: Chrysomelidae) under quarantine conditions in Australia. Native range surveys for additional potential biological control agents will also be pursued in northern and western Africa.

Biological control of bellyache bush (Jatropha gossypiifolia) in Australia: South America as a possible source of natural enemies

Bellyache bush (Jatropha gossypiifolia, Euphorbiaceae), a deciduous shrub introduced as an ornamental from tropical America, is a major and expanding weed of rangelands and riparian zones in northern Australia. Biological control is the most economically viable and long-term management solution for this weed. Surveys for potential biological control agents for J gossypiifolia in Mexico,Central America and the Caribbean resulted in release of the seed-feeding jewel bug Agonosoma trilineatum (Hemiptera: Scutelleridae), which failed to establish, and prioritisation of a leaf-rust Phakopsora arthuriana (Puccineales: Phakopsoraceae) for host-specificity testing, which is ongoing. With poor prospects for new agents from Mexico and Central America and the Caribbean, the search for candidate agents on J gossypiifolia shifted to localities south of the equator. Surveys were conducted on the purple-leaf form of J gossypiifolia, Jatropha excisa, Jatropha clavuligera and Jatropha curcas in Peru, Bolivia and Paraguay in 2012 and 2013. A total of 11 insect species, one mite species and the leaf-rust (P. arthuriana) were observed. These include a yet to be described leafmining moth (Stomphastis sp.) (Lepidoptera: Gracillaridae), a shoot and leaf-galling midge Prodiplosis longifila, and leaf-feeding midge Prodiplosis sp. near longifila (both Diptera:Cecidomyiidae) and an unidentified leaf-feeding moth larva (Lepidoptera: Notodontidae). The leafminer is widespread and damaging and has a field host range restricted to the genus Jatropha in Peru and Bolivia, holds the greatest promise as a biological control agent in Australia. Phakopsora arthuriana was recorded for the first time ever from Bolivia and Peru. Further exploration will be conducted in Peru and Bolivia during the wet season to confirm the field host range of collected agents,and to look for more new agents. Promising agents with field host-range restricted to Jatropha spp. will be imported into a quarantine facility in Australia for host-specificity testing.

Life cycle and host range of Phycitasp. rejected for biological control of prickly acacia in Australia

Prickly acacia (Vachellia nilotica subsp. indica), a native of the Indian subcontinent, is a serious weed of the grazing areas of northern Australia and is a target for classical biological control. Native range surveys in India identified a leaf webber, Phycita sp. (Lepidoptera: Pyralidae) as a prospective biological control agent for prickly acacia. In this study, we report the life cycle and host-specificity test results Phycita sp. and highlight the contradictory results between the no-choice tests in India and Australia and the field host range in India. In no-choice tests in India and Australia, Phycita sp. completed development on two of 11 and 16 of 27 non-target test plant species, respectively. Although Phycita sp. fed and completed development on two non-target test plant species (Vachellia planifrons and V. leucophloea) in no-choice tests in India, there was no evidence of the insect on the two non-target test plant species in the field. Our contention is that oviposition behaviour could be the key mechanism in host selection of Phycita sp., resulting in its incidence only on prickly acacia in India. This is supported by paired oviposition choice tests involving three test plant species (Acacia baileyana, A. mearnsii and A. deanei) in quarantine in Australia, where eggs were laid only on prickly acacia. However, in paired oviposition choice trials, only few eggs were laid, making the results unreliable. Although oviposition choice tests suggest that prickly acacia is the most preferred and natural host, difficulties in conducting choice oviposition tests with fully grown trees under quarantine conditions in Australia and the logistic difficulties of conducting open-field tests with fully grown native Australian plants in India have led to rejection of Phycita sp. as a potential biological control agent for prickly acacia in Australia.

The leaf-feeding geometrid Isturgia disputaria (Guenee)-A potential biological control agent for prickly acacia, Vachellia nilotica subsp. indica (Benth.) Kyal. & Boatwr. (Mimosaceae) in Australia

Prickly acacia (Vachellia nilotica subsp. indica), a native multipurpose tree in India, is a weed of National significance, and a target for biological control in Australia. Based on plant genetic and climatic similarities, native range surveys for identifying potential biological control agents for prickly acacia were conducted in India during 2008-2011. In the survey leaf-feeding geometrid, Isturgia disputaria Guenee (syn. Tephrina pulinda), widespread in Tamil Nadu and Karnataka States, was prioritized as a potential biological control agent based on field host range, damage potential and no choice test on non target plant species. Though the field host range study exhibited that V. nilotica ssp. indica and V. nilotica ssp. tomentosa were the primary hosts for successful development of the insect, I. disputaria, replicated no - choice larval feeding and development tests conducted on cut foliage and live plants of nine non-target acacia test plant species in India revealed the larval feeding and development on three of the nine non-target acacia species, V. tortilis, V. planiferons and V. leucophloea in addition to the V. nilotica ssp. indica and V. nilotica ssp. tomentosa. However, the proportion of larvae developing into adults was higher on V. nilotica subsp. indica and V. nilotica subsp. tomentosa, with 90% and 80% of the larvae completing development, respectively. In contrast, the larval mortality was higher on V. tortilis (70%), V. leucophloea (90%) and V. planiferons (70%). The no-choice test results support the earlier host specificity test results of I. disputaria from Pakistan, Kenya and under quarantine in Australia. Contrasting results between field host range and host use pattern under no-choice conditions are discussed.