A Morphometric Method to Assist in Defining the South American Origins of Microctonus hyperodae Loan (Hymenoptera: Braconidae) Established in New Zealand

In 1991, Microctonus hyperodae Loan (Hymenoptera: Braconidae, Euphorinae), collected from eight diverse South American locations, was released in New Zealand as a biological control agent of the pasture pest Listronotus bonariensis (Kuschel) (Coleoptera: Curculionidae). Equal numbers of each South American geographical population of this thelytokous endoparasitoid of adult hosts were released in New Zealand. A morphometric method described in this paper provided a means of classifying M. hyperodae adults as originating from either east or west of the Andes. Analysis revealed that, on average, specimens collected in Chile had significantly more antennal segments, narrower stigmas and longer radial cells than those collected in Brazil, Uruguay and Argentina. Significant differences between populations originating from each side of the Andes were also recorded in the dimensions of metasomal tergum one and basitarsus three. The morphometric method should, therefore, assist in defining the origins of M. hyperodae established in New Zealand. This information will be used in attempts to ascertain the importance of climate matching and host-parasitoid co-evolution to the outcome of the L. bonariensis biological control programme.     

Allozyme markers to help define the South American origins of Microctonus hyperodae (Hymenoptera: Braconidae) established in New Zealand for biological control of Argentine stem weevil

The thelytokous parasitoid, Microctonus hyperodae Loan, was collected from eight South American locations and introduced to New Zealand in 1991 for biological control of Argentine stem weevil, Listronotus bonariensis (Kuschel) (Coleoptera: Curculionidae). Parasitoids from each population were released in equal numbers at each New Zealand site to give them the same opportunities to establish. Population markers have been sought to identify the South American geographic populations that have become most successful in New Zealand. These would assist in determining the importance of concepts such as climate matching and host-parasitoid coevolution to the establishment of natural enemies in new regions for biological control. Vertical polyacrylamide electrophoresis was used to survey 16 enzymes and ten calcium binding proteins, and this paper reports variation at three putative loci. Malate dehydrogenase, a dihydrolipoamide dehydrogenase isozyme and a calcium binding protein exhibited clear genetic variation, each with two alleles. All M. hyperodae isofemale lines from east of the Andes mountains shared one genotype, all but one from west of the Andes shared another, while a population from within the Andes contained both genotypes. This variation was highly congruent with previously described morphometric variation. At two loci, the maintenance of heterozygotes, and the absence of homozygotes, within isofemale lines suggested M. hyperodae thelytoky is apomictic.     

Microsatellites and 16S sequences corroborate phenotypic evidence of trans-Andean variation in the parasitoid Microctonus hyperodae (Hymenoptera: Braconidae)

Eight South American geographical populations of the parasitoid Microctonus hyperodae Loan were collected in South America (Argentina, Brazil, Chile and Uruguay) and released in New Zealand for biological control of the weevil Listronotus bonariensis (Kuschel), a pest of pasture grasses and cereals. DNA sequencing (16S, COI, 28S, ITS1, beta-tubulin), RAPD, AFLP, microsatellite, SSCP and RFLP analyses were used to seek markers for discriminating between the South American populations. All of the South American populations were more homogeneous than expected. However, variation in microsatellites and 16S gene sequences corroborated morphological, allozyme and other phenotypic evidence of trans-Andes variation between the populations. The Chilean populations were the most genetically variable, while the variation present on the eastern side of the Andes mountains was a subset of that observed in Chile.     

Geographic Variation in Egg Load of Microctonus hyperodae Loan (Hymenoptera: Braconidae) and its Implications for Biological Control Success

Eight South American geographic populations of the parthenogenic, proovigenic, koinobiont parasitoid Microctonus hyperodae Loan (Hymenoptera: Braconidae: Euphorinae) were introduced to New Zealand to assist management of the pasture pest Listronotus bonariensis (Kuschel) (Coleoptera: Curculionidae). Geographic variation in fecundity has been suggested as a reason for the populations' differential successes in establishing in New Zealand. This study investigated whether geographic variation in fecundity was due to corresponding variation in pre-oviposition egg load (other possible sources of fecundity variation include searching efficiency, egg survival and female longevity). Variation in egg load accounted for that in fecundity, but also showed that the variation in fecundity was not as great as it had first appeared. Geographic variation in egg load did not explain the pattern of population establishment observed in New Zealand. Egg load was proportional to parasitoid size and this relationship was stronger in populations originating from west, rather than east, of the Andes. A new method for making parasitoid eggs visible is described.     

Early impact of endoparasitoid Microctonus hyperodae (Hymenoptera: Braconidae) after its establishment in Listronotus bonariensis (Coleoptera: Curculionidae) populations of northern New Zealand pastures

The South American curculionid Listronotus bonariensis (Kuschel) is an important pest of pastures in New Zealand. Population census data were gathered for L. bonariensis in northern New Zealand pastures during 1980-1983 in the absence of parasitism and again in 1991-1996 after the introduction and establishment of the braconid parasitoid Microctonus hyperodae Loan as a biological control agent. M. hyperodae achieved high rates of parasitism, with 75-90% of overwintering L. bonariensis parasitized within 3 yr of the parasitoid establishing at a site. Multistratum analysis of variance (ANOVA), with allowance for variation in host plant resource (numbers of Neotyphodium-free grass tillers), indicated reduction in the abundance of L. bonariensis life stages in the early part of life cycle. Although providing evidence for suppression of L. bonariensis, these analyses indicated the regulatory role of M. hyperodae was weak because L. bonariensis populations continued to exhibit marked intergenerational variability in abundance. Analyses of life tables indicated larval + pupal survival contributed most to intergenerational changes in abundance, irrespective of presence or absence of M. hyperodae. However, the density dependence of the stage survivals was modified in the presence of the parasitoid, with loss of density-dependent mortality in overwintering adults and increased density dependence in population natality. Regression analyses indicated dual contribution of parasitism and host plant resource to regulation of population natality and population trend in L. bonariensis. We conclude that M. hyperodae is a useful adjunct to host plant resistance in reducing the economic status of L. bonariensis populations in northern New Zealand pastures.     

Argentine stem weevil ( Listronotus bonariensis, Coleoptera: Curculionidae) population dynamics in Canterbury, New Zealand dryland pasture

The Argentine stem weevil (Listronotus bonariensis) was an economically important pest in New Zealand pastures until the release of the parasitoid Microctonus hyperodae. This contribution uses historical data to investigate the regulation of the pest populations prior to, and somewhat during, the establishment of this parasitoid in dryland Canterbury, New Zealand. Thus, a significant goal of this study is to provide an L. bonariensis population dynamics baseline for any future work that aims to analyse the full effects of M. hyperodae on the weevil, now that equilibrium with the weevil host has been reached.The population dynamics of L. bonariensis, based on a life-table approach, were investigated using data collected regularly for eight years from populations in Canterbury, New Zealand. The key factor affecting end-of-season L. bonariensis density was found to be variation in second generation fourth instar prepupal and pupal mortality. This may have been caused by arrested development and ongoing mortality resulting from the onset of cooler autumnal conditions.A compensatory response was found in recruitment to the second summer weevil generation, whereby the realised fecundity of the emergent first summer generation of weevils was found to be negatively related to the density of adult weevils per ryegrass tiller. This is the first time that this has been found via long-term population analysis of L. bonariensis, although indications of this have been found elsewhere in caging, pot and small plot experiments.In this study, the effect of the parasitoid biocontrol agent Microctonus hyperodae on L. bonariensis population dynamics was unclear, as the analysis covered a period when the parasitoid Microctonus hyperodae was introduced and still establishing. It does, however, raise important questions for future analysis in terms of the interaction between parasitism and unrealised fecundity.The results in this contribution also highlighted regional differences. Overwintering mortality of adult weevils in Canterbury was constant between years, whilst earlier studies in the North Island Waikato region indicated this mortality was density dependent. In addition, the availability of tillers in endophyte-free ryegrass pastures in Canterbury had no influence on egg and early-instar larval survival, which contrasts with the finding from endophytic Waikato pastures.     

Isolate-dependent impacts of fungal endophytes in a multitrophic interaction

Neotyphodium lolii , an endophytic fungus of perennial ryegrass, deters Argentine stem weevil, Listronotus bonariensis , an important insect pest of pastures in New Zealand. Deterrence is apparently due to several alkaloids the fungus produces. We asked if the fungus also affects the third trophic level. Specifically, we tested if several different isolates of the fungus altered the growth and survival of the parasitoid, Microctonus hyperodae . Adult weevils were collected from paddocks near Lincoln, New Zealand and maintained in an environmental chamber. Weevils were assorted into treatment groups and fed perennial ryegrass (cv. Nui) lacking endophyte infection (nil) or containing one of the following endophyte strains: ARW, AR1, AR6, AR37. All endophyte strains differed from one another with respect to the profile of alkaloids they produce. Following two weeks of acclimation, weevils were placed with M. hyperodae to allow for parasitization. Weevils were then placed into Petri dishes (10 per dish) and fed grass clippings from the appropriate treatment group. We collected weevils as they died and dissected them to assess the development of parasitoids. Emergence of prepupae from hosts and survival to adulthood were also recorded. Fungal isolates did not differ in their influence on weevil feeding or survival. In contrast, the effect of the endophyte on the parasitoid varied among isolates of the fungus. Isolates ARW and AR6 reduced parasitoid adult emergence relative to nil endophyte. In contrast, AR37 had no negative effect on survival of the parasitoid. Furthermore, an index of developmental rate showed that parasitoids developed more slowly when reared from hosts fed grass containing any of the strains of N. lolii , except AR37, compared to endophyte-free grass. Negative effects of the endophyte on parasitoid survival were associated with the presence of ergovaline while effects on parasitoid development rate were associated with the presence of any alkaloid.     

A prospective model for the phenology of Microctonus hyperodae (Hymenoptera: Braconidae), a potential biological control agent of argentine stem weevil in New Zealand

A predictive phenological model is described for the parasitoid Microctonus hyperodae, introduced to New Zealand as a potential biological control agent against Argentine stem weevil Listronotus bonariensis. The model is based on development/temperature relationships obtained from experiments on the parasitoid in quarantine prior to its release, allowing early predictions of its phenology in different parts of the target pest's New Zealand range. In particular the model was used to predict the number of parasitoid generations each year, the degree of temporal synchrony between parasitoid adults and the susceptible adult pest stage, the order of parasitism and reproduction in the pest's life cycle as a possible basis for a simplified, discrete host/parasitoid population model, and the likely significance of ecotypic differences in development and diapause characteristics of the parasitoid. These applications demonstrate the potential for simple models to help in climate matching of classical biological control agents and estimation of their interaction with pest dynamics, using data obtainable prior to their introduction and release. In addition the model proved useful as a decision aid during the release programme, by indicating the likely effects of unusual weather and the need or otherwise for further parasitoid releases.     

The biogeography of the austral, subalpine genus Ourisia (Plantaginaceae) based on molecular phylogenetic evidence: South American origin and dispersal to New Zealand and Tasmania

Molecular phylogenetic analyses of 26 of the 28 species of Ourisia , including eight of ten subspecies and two purported natural hybrids, are presented and used to examine the biogeography of the genus, which is distributed in subalpine to alpine habitats of South America, New Zealand and Tasmania. Gondwanan vicariance, often cited as the cause of this classic austral biogeographical pattern, was rejected by parametric bootstrapping of our combined dataset. Alternatively, various lines of evidence are presented in favour of a South American origin of Ourisia and subsequent dispersal to Australasia. Specifically, the genus likely arose in the Andes of central Chile and spread to southern Chile and Argentina, to the north-central Andes, and finally to Tasmania and New Zealand. The ancestor of the New Zealand species probably first arrived on the South Island, where the New Zealand species of Ourisia are most diverse, and migrated to the North and Stewart Islands. Because the Tasmanian and New Zealand species are sister to one another, the direction of dispersal between these two areas is equivocal. These results agree with other molecular phylogenetic studies that show that past dispersal between southern hemisphere continents has played an important role in the evolutionary history of many high-elevation austral plants. Our data also show that within South America, many of the geographical barriers (with the exception of the Atacama Desert) that have played a role in the evolution of other plant groups have not affected Ourisia species. Within New Zealand, the phylogeny and biogeography of species of Ourisia coincide with the geological history of the country and patterns of other alpine plants. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 479–513.     

Effect of container type on suitability of the pathogen Serratia marcescens - Microctonus hyperodae (Hym., Braconidae) association to indicate parasitoid oviposition attempts

To minimize the risk that introduced entomophagous biological control agents may pose to native and beneficial insect species, identification of likely nontarget hosts is required before release is considered. A novel technique, in which the parasitoid acts as a vector of a bacterium pathogenic to potential hosts, has been used as an indicator of parasitoid ovipositional activity even when subsequent development of the parasitoid egg within the 'host' does not occur. In this study the application of this technique was examined in relation to container type using the facultative bacterium Serratia marcescens in association with the parasitoid Microctonus hyperodae. Two permissive host weevil species, Listronotus bonariensis and Irenimus aequalis , were exposed for 72 h to M. hyperodae treated with either distilled water or S. marcescens suspension in either Petri dishes or cages. Weevil septicaemia was significantly higher when exposed to S. marcescens -treated parasitoids than water-treated parasitoids (P < 0.001), with no difference in mortality between weevil species. Septicaemia was significantly higher in the Petri dishes than in cages (P < 0.001). This was attributed to improved survival of S. marcescens in the Petri dishes compared with cages. Total parasitoid activity (combined septicaemia and parasitism) for I. aequalis was higher in Petri dishes than in cages and far exceeded field parasitism. Petri dishes provide an environment for constant interaction between parasitoid and target host and thereby provide a conservative indication of parasitoid host range. However, cages may be more suitable to conduct host range studies because the ability to manipulate the environment. Potential refinements to the parasitoid-bacteria technique include methodology that enhances viability of the bacterium under cage conditions.     

A Retrospective Analysis of the Establishment and Dispersal of the Introduced Australian Parasitoids Xanthopimpla rhopaloceros (Krieger) (Hymenoptera: Ichneumonidae) and Trigonospila brevifacies (Hardy) (Diptera: Tachinidae) within New Zealand

Two Australian parasitoids, Xanthopimpla rhopaloceros (Krieger) and Trigonospila brevifacies (Hardy), were introduced to New Zealand to control the light-brown apple moth (Epiphyas postvittana Walker). Dispersal by the parasitoids has since occurred naturally and with the aid of releases in fruit-growing areas. The present geographical range of the parasitoids includes all the North Island and some offshore islands to latitude 41 20 S. X. rhopaloceros is also present to latitude 41 48 S in the South Island. Comparisons of these distributions with those in Australia indicate that climatic conditions may have played a major role in the areas of establishment of both species in New Zealand. The mean winter temperature may be a limiting factor in the dispersal of T. brevifacies and X. rhopaloceros in New Zealand. Other factors that have probably aided the successful dispersal of the parasitoids include the wide distribution of host Tortricidae and the occurrence of tortricid host plants. The areas of New Zealand that appear suitable for further colonization by T. brevifacies include northern areas of the South Island, and both parasitoids could disperse further into suitable climate areas of the east and west coasts of the central South Island. The rate of dispersal for X. rhopaloceros was estimated at 13-24 km/year, and for T. brevifacies at 8-15 km/year.     

The galaxiid fishes of South America

Four galaxiid species in two genera are recognized and redescribed from the South American region, viz., Galaxias maculatus (Jenyns), G. platei Steindachner, G. globiceps Eigenmann, and Brachygalaxias bullocki (Regan). All of these species occur in Chile, but G. maculatus and G. platei occur also east of the Andean divide in Argentina, in Tierra del Fuego, and the Falkland Islands. The two species east of the Andes are regarded as having crossed the mountains from west to east. The galaxiids, together with Aplochiton (family Aplochitonidae) and Geotria (family Petromyzonidae), are regarded as southern circumpolar elements in the South American freshwater fish fauna, distinct from the group of species of Amazonian derivation (trichomycterids, Nematogenys, Diplomystes, Cheirodon , etc.) and distinct also from oceanic derivatives, either recent (atherinids) or ancient (serranids).     

Chloroplast DNA variation in the Turnera sidoides L. complex (Turneraceae): biogeographical implications

Aim To analyse the current geographical structure of chloroplast DNA variation in the Turnera sidoides L. complex in order to establish historical biogeographical hypotheses for the mid‐latitude South American lowlands. During the Quaternary, the climate shifted from tropical humid to cold dry, and the vegetation cover has not been stable. The consequences of these processes on the current distribution of the vegetation of this area have received very little attention. Location The mid‐latitude South American lowlands extend between c. 20 and 40°S and include Uruguay, northern, central and eastern Argentina, southern Brazil, and parts of southern Paraguay and Bolivia. They are surrounded by higher‐elevation systems. Methods Turnera sidoides is a well‐studied polyploid complex of perennial rhizomatous herbs occurring throughout the area of interest. We analysed 321 individuals from 79 populations of the five recognized subspecies. We also included progenies from artificial crosses in order to analyse chloroplast inheritance. After screening sequences for four non‐coding chloroplast DNA regions, the trnL–trnF spacer was selected to characterize the collection. Results Three haplotypes can be easily identified, with each differing from the others in two independent characters. A clear geographic structure is revealed when haplotypes are plotted for the complex as a whole regardless of subspecies and cytotype. Three distinct regions can be identified. Main conclusions We propose three putative refugial areas for the Turnera sidoides complex, which are associated with the orographical systems of the region. Ravines and slopes in the Haedo Cuchilla system in northern Uruguay, the elevations of the western side of the area in Argentina, and the eastern Serranías system in south‐eastern Uruguay may each have served as refugia in which the A, B and C haplotypes became fixed during the drier climatic phases. Biogeographical patterns in the area covered by T. sidoides, particularly east of the Uruguay River, have not previously been analysed from a historical perspective.     

Evidence of Host Discrimination by Microctonus hyperodae Loan (Hymenoptera: Braconidae, Euphorinae), a Parasitoid of the Argentine Stem Weevil Listronotus bonariensis (Kuschel) (Coleoptera: Curculionidae)

Microctonus hyperodae is a solitary endoparasitoid of the Argentine stem weevil, Listronotus bonariensis. Early investigation into the biology of the parasitoid indicated that there was no discrimination between parasitized and unparasitized hosts. However, dissection data from two experiments were analyzed according to three mathematical models. Model I was based on the Poisson distribution and assumed random selection of hosts. The host discrimination model (model II) assumed that parasitized hosts had reduced attractiveness to searching parasitoids. A competition model (model III) made the assumption that competition between the early immature parasitoid stages led to premature mortality, which was not accounted for in the dissection results. The dissection data indicated statistically significant departure from the Poisson model. Results from the laboratory experiment indicated that host discrimination explained the results more accurately than parasitoid larval competition. Models II and III both provided adequate fits to the field data, although the departure from the observed data was greatest for the competition model. Both intra- and inter-ecotypic host recognition were evident, and there was some indication that clonal recognition took place. This is the first evidence of host discrimination by a member of the genus Microctonus. It has provided further indication of the success of M. hyperodae as a biological control agent.     

Low genetic differentiation between two geographically separated populations of demersal gadiform fishes in the Southern Hemisphere

The distribution patterns of many fishes between the three continents (Africa, Australia, and South America) in the Southern Hemisphere have been uncovered to be influenced by mostly vicariance or historical dispersal. Although some demersal fishes with intercontinental distribution are suggested to be more influenced by current/recent dispersal, few genetic studies have been made for demersal fishes so far. To provide more information for such fishes, genetic divergence was analyzed for two pairs of gadiform species and subspecies distributed around Australasia and South America: the blue grenadier, Macruronus novaezelandiae (from New Zealand) and the Patagonian grenadier, M. magellanicus (from South America) as well as two subspecies of the southern blue whiting, Micromesistius australis pallidus (from New Zealand) and M. a. australis (from South America). The sequence analyses of two mitochondrial DNA regions showed no divergence between Australasian and South American populations of the grenadiers and the southern blue whiting. The microsatellite DNA analysis also indicated significant but very minimal genetic differentiation between the two geographic populations of each pair. These results imply rather recent separation of the two geographic populations. Current/recent dispersal may be an important common factor for determining the distribution of demersal fishes in the Southern Hemisphere. Nonetheless, low but significant genetic differentiation observed requires treating the two populations of the economically important grenadiers and southern blue whiting, respectively, as different stocks for proper resource management.     

Review of the genus Hippidion Owen, 1869 (Mammalia: Perissodactyla) from the Pleistocene of South America

This review of Hippidion is based on a multivariate analysis of the foot, and some morphological characteristics of the skull and dentition. We recognize only one genus ( Hippidion ) including all the hippidiform horses, with three different species: H. principale, H. devillei and H. saldiasi. The latter species is stratigraphically and geographically restricted to the period from 13000 to 8000 years BP in the southern part of South America. Hippidion principale and H. devillei have a large geographical distribution (Argentina, Bolivia, Chile, Perú, Uruguay, Brazil) through the Upper Pliocene-Upper Pleistocene. Both species show some morphometric variations across their geographic range; these features may result from the environmental characteristics.     

Behavioural Acceptability of Sitona lepidus (Coleoptera: Curculionidae) to the Parasitoid Microctonus aethiopoides (Hymenoptera: Braconidae) Using the Pathogenic Bacterium Serratia marcescens Bizio

Microctonous aethiopoides Loan has been introduced into New Zealand to control the lucerne pest Sitona discoideus Gyllenhal (Coleoptera: Curculionidae). Sitona lepidus Gyllenhal (Coleoptera: Curculionidae) a pest of clover ( Trifolium spp.), has recently established in New Zealand. Laboratory experiments to test the potential of M. aethiopoides to parasitize S. lepidus has resulted in very low levels of parasitism. To investigate whether there were behavioural or physiological barriers to successful parasitism, two experiments were conducted using the insect pathogenic bacterium. Serratia marcescens Bizio as a marker for parasitoid ovipositor penetration. Firstly, M. aethiopoides 'treated' with S. marcescens were exposed to weevils and rapid weevil mortality was used to indicate ovipositor penetration. Up to 50% mortality of S. lepidus occurred, which was comparable with mortality observed in the permissive host Listronotus bonariensis . Dissection of S. lepidus exposed to parasitoids treated with distilled water showed that ca. 21% contained parasitoid eggs of which 98% were nonviable. In the second experiment, exposure periods of 24, 48 and 72 h to S. marcescens -treated parasitoids produced an increase in S. lepidus mortality of 14, 28 and 38%, respectively. There was 3% successful parasitoid development in weevils exposed for 72 h to parasitoids treated with distilled water. M. aethiopoides has been shown to develop successfully in a wide range of non-target weevil species both in the laboratory and field. Possible reasons for poor survival of M. aethiopoides immature stages in S. lepidus are discussed.     

Spread and impact of introduced conifers in South America: Lessons from other southern hemisphere regions

The history of conifers introduced earlier elsewhere in the southern hemisphere suggests that recent invasions in Argentina, Brazil, Chile and Uruguay are likely to increase in number and size. In South Africa, New Zealand and Australia, early ornamental introductions and small forestry plantations did not lead to large‐scale invasions, while subsequent large plantations were followed with a lag of about 20–30 years by troublesome invasions. Large‐scale conifer plantation forestry in South America began about 50–80 years later than in South Africa, Australia and New Zealand, while reports of invasions in South America lagged behind those in the latter nations by a century. Impacts of invading non‐native conifers outside South America are varied and include replacement of grassland and shrubland by conifer forest, alteration of fire and hydrological regimes, modification of soil nutrients, and changes in aboveground and belowground biotic communities. Several of these effects have already been detected in various parts of South America undergoing conifer invasion. The sheer amount of area planted in conifers is already very large in Chile and growing rapidly in Argentina and Brazil. This mass of reproductive trees, in turn, produces an enormous propagule pressure that may accelerate ongoing invasions and spark new ones at an increasing rate. Regulations to control conifer invasions, including measures to mitigate spread, were belatedly implemented in New Zealand and South Africa, as well as in certain Australian states, inspired by observations on invasions in those nations. Regulations in South America are weaker and piecemeal, but the existing research base on conifer invasions elsewhere could be useful in fashioning effective regulations in South America. Pressure from foreign customers in South Africa has led most companies there to seek certification through the Forestry Stewardship Council; a similar programme operates in Australia. Such an approach may be promising in South America.     

Should the flower bud weevil Anthonomus santacruzi (Coleoptera: Curculionidae) be considered for release against the invasive tree Solanum mauritianum (Solanaceae) in New Zealand?

The invasive tree Solanum mauritianum Scop. has been targeted for biological control in South Africa and New Zealand, by deploying insect agents that could constrain its excessive reproductive output. The flower-feeding weevil Anthonomus santacruzi (Curculionidae) was approved for release in South Africa in 2007 but following the loss of the original culture in quarantine, new stocks were introduced from Argentina in 2008–2009. This study was initiated to confirm that the host range of the new culture was the same as that of the previous one, but also to assess the risks associated with the weevil's release in New Zealand. Different testing procedures, including no-choice tests and multi-choice tests in different arenas, produced inconsistent and ambiguous results. During no-choice tests, oviposition and larval development to adulthood occurred on three non-target species including two native South African and one native New Zealand Solanum species. However, subsequent multi-choice tests and a risk assessment suggested that the risks of anything more than collateral damage to non-target Solanum species are low. Overall, these data do not deviate substantially from the results of the original quarantine tests which facilitated the release of A. santacruzi in South Africa in 2009. Although we argue that none of the New Zealand native and cultivated species are at risk, stronger evidence from open-field trials and chemical ecology studies may be required to convince the regulatory authorities that A. santacruzi is suitable for release in New Zealand.