Virus-like particles (MaVLP) have been discovered in the ovarial epithelial cells of the solitary, koinobiont, endoparasitoid, Microctonus aethiopoides Loan (Hymenoptera: Braconidae) introduced to New Zealand originally from Morocco to control the lucerne pest Sitona discoideus Gyllenhal (Coleoptera: Curculionidae). MaVLP have been found in all females examined. It has been suggested, although not demonstrated, that like many other such VLP found in parasitoids, MaVLP might play a role in host immunosuppression. Since another biotype of M. aethiopoides from Ireland has been proposed for introduction to control the white clover pest, Sitona lepidus Gyllenhal, in New Zealand, it was considered that females from this biotype warranted transmission electron microscope examination for VLP. No VLP were observed in ovarian tissues of specimens collected from three different locations in Ireland. Similarly, none were found in M. aethiopoides sourced from France, Wales, and Norway. These observations are discussed in relation to quarantine host specificity tests with the Irish biotype, which found that the host range of the Irish biotype is likely to be less extensive than that of the Moroccan biotype already in New Zealand.

The braconid parasitoid Microctonus aethiopoides Loan has been released in Australia and New Zealand for biological control of the lucerne pest Sitona discoideus Gyllenhal. In New Zealand, the parasitoid attacks a number of endemic weevil species. A survey of Curculionoidea found in and near lucerne in south-eastern Australia was carried out to investigate whether similar non-target parasitism was occurring, and to relate this to levels of parasitism found in the target host, S. discoideus. Some of the original M. aethiopoides release sites were particularly targeted in the survey of 25 sites in Victoria, New South Wales and South Australia. Almost 2500 weevils were collected, of which over 90% were S. discoideus, with the remaining 197 other weevils comprising 29 species found at 15 of the 25 sites. Parasitism of S. discoideus by M. aethiopoides occurred at 12 lucerne sites, with levels ranging from 0 to 25%. A single incidence of parasitism of a species of an Australian native weevil Prosayleus sp. by M. aethiopoides was recorded. No parasitism of any other weevil species was observed. The taxonomic affinities between Sitona and native Australian and New Zealand weevils are discussed, concluding that non-target host range in M. aethiopoides may be determined more by ecological factors than by taxonomic affinities among its hosts.

Abstract This contribution introduces a short series of papers on biodiversity of indigenous tussock grassland at four sites in New Zealand. A survey of invertebrates of tussock grassland sites was carried out in summer of 3 successive years, 2001–03. The sites included two in Otago in snow tussock grassland at Deep Stream and Mt Benger, a site at Cass in inland Canterbury, and one at Tukino in the central North Island. Sampling was carried out by taking turf samples from each site, and extracting the invertebrates with heat extractors. The invertebrates were divided into 30 major groups. This quantitative sampling method allowed the densities of invertebrate groups to be calculated. Total macro‐invertebrate density for all sites ranged between c. 1600 and 5600 m–2. Total invertebrate abundance was highest at the central North Island site, but this was very much dominated by Formicidae. Across all sites, Formicidae, Coleoptera, and Pseudococcidae were consistently the most abundant groups represented.

The European biotype of the parasitoid Microctonus aethiopoides Loan ( Hymenoptera: Braconidae) is being considered for release against Sitona lepidus Gyllenhal ( Coleoptera: Curculionidae) in New Zealand. Host specificity was evaluated in the laboratory using both endemic and introduced weed biological control curculionid species, with 12 no-choice and three choice experiments carried out comparing the S. lepidus and test weevils. Two further no-choice tests used the Moroccan M. aethiopoides biotype to compare attack rate between European and Moroccan M. aethiopoides, the latter released in 1982 to control the lucerne pest S. discoideus. Across all experiments, total parasitism of S. lepidus was 69% compared with 15% for the test weevils. European M. aethiopoides was able to develop in the native weevils Irenimus aequalis, Nicaeana cervina, Catoptes cuspidatus, Protolobus porculus and Steriphus variabilis with parasitism rates of 13, 28, 2, 7 and 8%, respectively. These levels were significantly less than those in the corresponding S. lepidus control. Total parasitism of I. aequalis and C. cuspidatus increased significantly in the presence of S. lepidus than recorded under no-choice conditions. The presence of European M. aethiopoides caused minor, if any, test weevil mortality prior to the onset of prepupal emergence and there was no significant reproductive suppression in parasitoid-exposed test weevils. Parasitism of the introduced weed control agent R. conicus by European M. aethiopoides was significantly lower (1.1%) compared to the Moroccan biotype (47.5%). Based on these and other experiments, should the European M. aethiopoides be released as a biological control agent of S. lepidus, its ecological impacts are likely to be less severe than those already exhibited by the Moroccan M. aethiopoides.

Bacterial and fungal communities in indigenous tussock grassland soils were studied at four locations, Mt Benger and Deep Stream (Otago), Cass (Canterbury) and Tukino (central North Island). Soil samples collected from inter-tussock and tussock areas were used to enumerate total culturable bacteria (colony forming units (CFU)) as well as a number of specific groups of bacteria, fungal types and diversity, and soil microbial functional diversity. Soil microbial biomass carbon (C) and nitrogen (N) as well as extractable C and N were also determined. Fungal populations were lowest at Mt Benger while fluorescent Pseudomonas was lowest at Cass. In these indigenous soils, bacterial CFU were c. 30 times lower and fungal populations c. 10 times greater than in developed New Zealand pastoral soils. CFU of bacteria in complex media (r-strategists) were similar in magnitude to bacteria growing on weak CA medium (K-strategists). The microbial biomass C in indigenous grassland soils were also significantly greater than that found in developed pastoral soils, possibly due to dominance of fungi in indigenous soils. Microbial C and N were lower in Cass and Tukino sites compared to Mt Benger and Deep Stream sites. Microbial nitrogen was also significantly greater (P < 0.001) in inter-tussock than under tussock samples. The functional diversity of soil microbes, was significantly greater (P < 0.01) in inter-tussock samples compared to tussock samples at all sites except Mt Benger. The total microbial activity, as measured by the colour development in Biolog plates (Average Well Colour Development (AWCD)), was low in Cass soil compared to others. Eighty-three types of fungi belonging to 30 genera were recovered from the four sites. The diversity of fungi found in Otago sites was greater than in the other two sites (P < 0.001). Fusarium spp. that are often common in pastoral soils were conspicuous by their rarity in indigenous soils.

A method is described for estimating the impact of a parasitoid on the abundance of a nontarget host, using the intrinsic rate of host increase, the average abundance of the host in the presence of parasitism, and the estimated mortality caused by the parasitoid. The method is applied to the braconid Microctonus aethiopoides Loan, introduced to New Zealand to control Sitona discoideus Gyllenhal in lucerne but also attacking native weevils Irenimus spp. and Nicaeana spp. The nontarget host population was modeled using discrete Ricker or continuous logistic models, tuning the models to host population data in the presence of parasitism, then removing parasitism and determining the increase in predicted equilibrium host density. In an area where up to 30% parasitism of a nontarget host population has been recorded, the model estimated an 8% reduction of the nontarget host. In another area, where the parasitoid has not established, the method was applied in reverse to predict the parasitoid's impact if it did establish. In this case, the model predicted a 30% suppression of population density. The host's intrinsic rate of increase, r(m), accounts for this difference in predicted impact, which was small in the low altitude area where r(m) was high, and the impact was larger in the higher altitude site where r(m) was smaller.

Biodiversity studies are often limited by unavailability or inaccessibility of taxonomic expertise; in New Zealand, taxonomic revisions and keys to many invertebrate groups are far from complete. To make progress with ecological and biodiversity studies, the separation of organisms into recognisable taxonomic units or morphospecies has sometimes been adopted. Coleoptera are speciose, trophically diverse, and taxonomically well known compared with other large trophically diverse groups and so they are useful to include in biodiversity studies. This study opportunistically examined the accuracy of Coleoptera species separation using morphologically recognisable features of specimens collected from three different vegetation communities, by three student researchers with different levels of training and previous expertise. Their morphospecies were examined by a single researcher with experience in taxonomy of Coleoptera. In total, 155 morphospecies were separated by the three students, compared with 151 determined by the specialist, which included representatives from 23 families of Coleoptera. All three students identified a total number of morphospecies within about 10% of the actual number, irrespective of previous training. However, the proportion of correct species separation increased from 63 to 87% in accordance with the level of previous experience. Common errors in species separation made by parataxonomists in relation to coleopteran families are discussed.

Abstract Invertebrates exhibit exceptional levels of diversity and endemism in New Zealand where, historically, they have received only limited consideration in land management decisions. Many species exist outside the habitats typically set aside for conservation, such as lowland to subalpine Chionochloa tussock grasslands. These habitats are under‐represented as protected areas due to their modified state and their invertebrate fauna is poorly understood. Compiling inventories has been suggested as one means of facilitating a greater awareness of invertebrate diversity and ecology. This study presents an inventory of Curculionoidea recorded during a single quantitative sampling event in mid summer 2001, from two Otago Chionochloa tall‐tussock grasslands. Species diversity is compared with that of other southern South Island tussock grassland areas, and notes on weevil ecology and distribution are given. Of the 35 species known from the two sites, only 17 were recorded from samples taken in January 2001, demonstrating the importance of factors such as seasonality and microhabitat to study design. Genera recorded showed affinities with those of grassland studies in neighbouring ecological districts. Almost 50% of the species collected from the two sites were undescribed; this not only limits the capability of land managers to compare areas under consideration for protection or other land uses, but also indicates an abundance of unrecorded and unprotected biological diversity.

The yellow flower wasp Radumeris tasmaniensis Saussure was first reported in Northland New Zealand in February 2000 R tasmaniensis is a solitary wasp which occurs naturally in Australia and Papua New Guinea It is an ectoparasitoid of scarabaeid larvae A survey was carried out during February and March 2001 to determine the distribution and potential host range of R tasmaniensis in Northland This confirmed that R tasmaniensis was present at the three sites from which it was first reported in 2000 but appeared not to have established more widely A small extension to its known range was discovered in a further survey in MarchApril 2002 Parasitised scarabaeid larvae were not detected by soil sampling but Pericoptus spp (Scarabaeidae Dynastinae) was the species most commonly found The rationale and methodology of the survey is presented and the conservation implications of the establishment of this species in New Zealand are discussed

This study assessed the effects of host plant volume on invertebrate density and taxon richness on a native New Zealand shrub, Olearia bullata (Asteraceae). Specimens were collected by beating during a single summer/autumn sampling event from 30 O. bullata plants. The influence of the surrounding environment was also examined by assessing canopy connectivity between O. bullata shrubs and their nearest neighbours (O. bullata and other shrub species). Linear regression analyses suggested a positive association between invertebrate taxon richness and shrub volume, a relationship that was also apparent at the Order level for Coleoptera, Diptera, and Psocoptera. No significant results were found between shrub volume and invertebrate density at any taxonomic level. Negative associations were obtained between the density of Araneae and Hemiptera against the distance between O. bullata shrubs.

In times of biodiversity crisis there is an increasing need for faster and cheaper methods by which to achieve conservation goals. This situation is especially troublesome for invertebrates, and the use of morphospecies instead of taxonomic species has been proposed as a way around the taxonomic constraints in particular situations. We conducted a study in a modified native shrubland on New Zealand's South Island in which we sampled Lepidoptera, Coleoptera, and Araneae in autumn by beating and pitfall traps. All specimens were separated into morphospecies by a nonspecialist and identified by specialized taxonomists, and the results were compared. Results were analyzed with respect to correct separations (one taxonomic species to one morphospecies), lumping (more than one species classified as a single morphospecies), and splitting (one species separated into more than one morphospecies). Among the individual orders, Lepidoptera yielded the most accurate results (91% correct separation), whereas Coleoptera and Araneae yielded poor results (63% and 50%, respectively). The overall difference between the morphospecies and taxonomic species estimates for the site was only 3.3%, but this was an artifact caused by the splitting and lumping results balancing each other out. The accuracy of morphospecies separation varies greatly among different invertebrate groups, so the relationship between morphospecies and taxonomic species for a particular target group must be established beforehand. We recommend that some prior orientation should be given by expert taxonomists. When adopted with care, morphospecies present a useful tool for conservation, particularly for environmental impact assessment and when inventorying diversity does not require information on particular species.

Microctonus aethiopoides was first recorded parasitising the weed biocontrol agent Rhinocyllus conicus in 1994 Subsequent studies found parasitism at several sites in Otago and South Canterbury In this study a further 15 sites in Otago and South Canterbury were sampled to determine the extent and level of parasitism of R conicus by M aethiopoides Rhinocyllus conicus adults were collected from nodding thistles and were either reared or dissected to determine parasitism levels For dissected weevils the stages of parasitoid development presence of teratocytes weevil size and female weevil reproductive status were recorded Parasitism was recorded at 12 sites Lucerne the host plant of the weevil Sitona discoideus for which M aethiopoides was originally introduced into New Zealand was absent from most of these sites Levels of parasitism were similar to those recorded in previous studies reaching up to 17 in the Hakataramea Valley Results relating to weevil size and female weevil reproductive state are discussed

A laboratory study investigated development of teratocytes derived from the parasitoid Microctonus aethiopoides Loan in the natural host, Sitona discoideus Gyllenhal, and in three novel hosts, the introduced weed biological control agent Rhinocyllus conicus (Froehlich), and two New Zealand native species Nicaeana cervina Broun and Irenimus stolidus Broun. Weevils were exposed to parasitoids and then examined 6, 10 and 15 days post-parasitism for parasitoid stage and size, and teratocyte number and size. In all hosts, teratocyte numbers decreased and size increased as parasitoid development progressed, although 6 days after parasitism, fewer, larger teratocytes were found in I. stolidus than S. discoideus or N. cervina. In weevils containing second-third instar parasitoid larvae, the most permissive hosts, S. discoideus and N. cervina contained more teratocytes than the least permissive hosts I. stolidus and R. conicus. Host gender influenced some aspects of parasitoid and teratocyte development. Total teratocyte volume was greater in female than male S. discoideus at all sampling times, and at 10 days post-parasitism in N. cervina. A possible relationship between host suitability and teratocyte development is discussed.

This is the first published invertebrate survey focusing on a low-altitude shrubland community in New Zealand. Invertebrates were collected from a remnant native shrubland (450 m) protected by the Brookdale Conservation Covenant, Rock and Pillar Range, Otago, New Zealand in late summer/autumn 1999. Sampling was carried out by beating 30 randomly chosen shrubs of each of two native species: Olearia bullata H. D. Wilson & Garnock-Jones (Asteraceae) and Coprosma propinqua A. Cunn. (Rubiaceae). Fifty pitfall traps were also set under the same shrubs and on nearby open patches of exotic grassland. Three Phyla, six Classes, 25 Orders and approximately 280 species were recorded. An annotated list of taxa is presented, and plant/host associations plus other observations on the fauna are discussed. Approximately 90% of the identified species were endemic, emphasising the importance of such remnant habitats for the protection of New Zealand's biodiversity.

In rabbit-prone areas of the South Island of New Zealand, a field survey was undertaken of the incidence, relative abundance and seasonality of possible rabbit haemorrhagic disease virus (RHDV) vectors, blowflies (Diptera: Calliphoridae) and fleshflies (Diptera: Sarcophagidae). Flies were trapped over a 4-day period each month for a year except during the winter months. Liver-baited Western Australian fly traps were set in Marlborough (three sites), the Mackenzie Basin (three sites) and Central Otago (six sites). Eight species of calliphorids (blowflies) and the sarcophagid (fleshfly), Hybopygia varia (Walker) were trapped. Numbers were generally higher at shrubby or grassy sites compared with bare open sites with little vegetation. More H. varia were trapped at sites grazed by cattle than at other sites. In March and November 1999, and January 2000, sticky traps attached to cages containing either a dead, live, or no rabbit were set up at two of the Central Otago sites where rabbit control had been poor (Sugar Loaf), and good (Locharburn), respectively. Blowflies were more abundant on sticky traps at the Locharburn site than at Sugar Loaf. Both groups of flies were attracted in significantly greater numbers to dead rabbits compared with live rabbits and empty cages. No evidence of RHDV was found on flies from either site, on any of the three dates. The implications of these results are discussed in the context of rabbit biological control.

A laboratory study of aspects of parasitoid host acceptance, suitability and physiological regulation in natural and novel host species was carried out to investigate the degree of variability encountered with different hosts and to determine the value of such observations in host range determination. The parasitoid Microctonus aethiopoides Loan was exposed to a natural host, Sitona discoideus Gyllenhal (Coleoptera: Curculionidae) and three novel hosts, the New Zealand native Nicaeana cervina Broun, the introduced weed biological control agent Rhinocyllus conicus (Froehlich), and a congeneric pest species, Sitona lepidus Gyllenhal (all Coleoptera: Curculionidae). Per cent parasitism of these species was 54%, 43%, 39% and 0%, respectively. The results indicated that for both S. discoideus and R. conicus more males than females were parasitized (69% cf. 45%, and 49% cf. 32% respectively) but host size was not a significant factor. Overall, superparasitism was recorded in about 29% of parasitized weevils and there was evidence that host discrimination to avoid superparasitism occurred in the natural host. Conversely, superparasitism occurred more frequently than would be expected in N. cervina (42%) coupled with higher survival of larvae in superparasitized hosts in this species. The frequency distribution of attack of R. conicus by M. aethiopoides was not different from random. Parasitoid development was more rapid in the natural host, S. discoideus, and parasitoid size was positively correlated with host size. There was a strong positive relationship between parasitoid larval survival and the presence of teratocytes in all hosts. Host fecundity and fertility were reduced by parasitism for most species, and in some cases, by exposure to parasitoids in the absence of detectable parasitism. It was concluded that laboratory observations can provide useful information on the compatibility between host and parasitoid which can complement traditional host range tests to predict field host range.

Entomologia Experimentalis et ApplicataVolume 95, Issue 2 p. 213-216 Effects of Microtonus aethiopoides parasitism on hemolymph protein composition across alternate hosts Naomi Lovallo, Naomi Lovallo Department of Entomology, The Pennsylvania State University, 501 Agricultural Sciences and Industries Building, University Park, PA 16802, USA (Phone: (814) 865-3345; E-mail: [email protected])Search for more papers by this authorB.I.P. Barratt, B.I.P. Barratt AgResearch, Invermay Agricultural Centre Puddle Alley, Private Bag 50034 Mosgiel, New ZealandSearch for more papers by this authorM. Legge, M. Legge Department of Biochemistry, University of Otago, 676 Cumberland Street, Dunedin, New ZealandSearch for more papers by this authorD.L. Cox-Foster, D.L. Cox-Foster Department of Entomology, The Pennsylvania State University, 501 Agricultural Sciences and Industries Building, University Park, PA 16802, USA (Phone: (814) 865-3345; E-mail: [email protected])Search for more papers by this author Naomi Lovallo, Naomi Lovallo Department of Entomology, The Pennsylvania State University, 501 Agricultural Sciences and Industries Building, University Park, PA 16802, USA (Phone: (814) 865-3345; E-mail: [email protected])Search for more papers by this authorB.I.P. Barratt, B.I.P. Barratt AgResearch, Invermay Agricultural Centre Puddle Alley, Private Bag 50034 Mosgiel, New ZealandSearch for more papers by this authorM. Legge, M. Legge Department of Biochemistry, University of Otago, 676 Cumberland Street, Dunedin, New ZealandSearch for more papers by this authorD.L. Cox-Foster, D.L. Cox-Foster Department of Entomology, The Pennsylvania State University, 501 Agricultural Sciences and Industries Building, University Park, PA 16802, USA (Phone: (814) 865-3345; E-mail: [email protected])Search for more papers by this author First published: 07 October 2003 https://doi.org/10.1046/j.1570-7458.2000.00660.xCitations: 1AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume95, Issue2May 2000Pages 213-216 RelatedInformation

The phenology of native brachycerine weevil species at seven pasture sites in Otago, Canterbury and Waikato was studied by regular quantitative sampling of adults. Weevils were identified to species, and dissected to record reproductive status and parasitism by introduced braconid parasitoids in the genus Microctonus. Climatic data assisted in the interpretation of some population density patterns. Weevil population density was estimated for periods of two to five years at the selected sites. Species in the Entimini (species of Irenimus and Nicaeana) were generally univoltine, with adults emerging in winter-spring. The main period of reproductive activity was spring, and parasitism by Microctonus aethiopoides reached its highest incidence in January. Low level parasitism of native weevil species by M. aethiopoides was detected at all sites, and by M. hyperodae at two sites. At one site in Otago, parasitism by M. aethiopoides was higher and could have affected the population density of Irenimus aemulator (Broun) and Nicaeana sp. Most parasitism occurred after the main reproductive period of weevils in spring, but a putative second generation in some species might be more affected by parasitoid attack. A native rhytirhinine species, Steriphus variabilis, differed from the entimines because adults emerged in autumn and spring, and may be bivoltine. Mechanisms of M. aethiopoides parasitism of non-target species in the field are discussed.

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.

General biogeographic features of the two Ecological Districts - Umbrella c. 150 000 ha and Nokomai, c. 110 000 ha - of the Waikaia Ecological Region, south-central South Island, are described. Results of normal and inverse cluster analyses of plot samples of the full range of indigenous vegetation remaining in each District are presented. Up to 17 plant communities from each District are characterised in terms of physiognomy, flora, and physiography. These range from beech forests and lowland red tussock grassland through upland shrublands, shrub-tussocklands, tussocklands, sedgelands, and wetlands to high-alpine communities (snowbank, cushionfield, scree).The 649 indigenous and 97 adventive vascular plant taxa plus 21 hybrids are listed by District. Plant distributions, particularly altitudinal and geographic limits for many alpine and some threatened taxa, are described.Of the region's fauna, 61 birds, 268 Lepidoptera, 202 Coleoptera (some in both orders undescribed), and several other invertebrates including the rare land snail Powelliphanta spedeni spedeni are listed by District. Distributions of several rare andi or local taxa are described both within and beyond the Region. Aquatic fauna of unmodified upland lakes and ponds of Umbrella District are recorded.Within the Region, 31 areas ranging from 25 ha to 2620 ha are recommended for protection, 20 (6260 ha or 2.4%) from the Umbrella District and 11 (11 615 ha or 9.5%) from the Nokomai District. These were selected so as to adequately represent the full range of remaining indigenous ecosystems and their associated landforms.Details of progress (to July 1997) with the implementation phase of the programme are outlined. Of the 31 Recommended Areas for Protection (RAPs) identified in the field surveys, seven now have some degree of protection over their full extent and nine are partly protected. These protected areas represent 44% (c. 7985 ha) of the total area of the 31 RAPs (18 060 ha) identified. Tenure review of pastoral leasehold land continues to be an important means of protecting conservation values in these two Districts, as in other parts of the South Island high country.