T.L. Nelson

Yersinia entomophaga sp. nov., isolated from the New Zealand grass grub Costelytra zealandica

A Gram-negative, rod-shaped, non-spore-forming bacterium (MH96(T)) was isolated from diseased larvae of the New Zealand grass grub, Costelytra zealandica (Coleoptera: Scarabaeidae). On the basis of 16S rRNA gene sequence similarity, strain MH96(T) is a member of the genus Yersinia, which is a member of the class Gammaproteobacteria. The most similar 16S rRNA gene sequence to that of MH96(T) is that of the type strain of Yersinia mollaretii (98.5 % similarity) followed by those of the type strains of Yersinia aldovae, Y. frederiksenii and Y. rohdei (all 98.4 % similarity). Multilocus sequence typing of five housekeeping genes (dnaJ, glnA, gyrB, groEL and recA) identified Yersinia ruckeri (81-92 % similarity) as the closest relative. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain MH96(T) from the four most closely related Yersinia species with validly published names, including a Y. ruckeri isolate. Strain MH96(T) therefore represents a novel species, for which the name Yersinia entomophaga sp. nov. is proposed, with the type strain MH96(T) ( = DSM 22339(T)  = ATCC BAA-1678(T)).

Beauveria caledonica is a naturally occurring pathogen of forest beetles.

In New Zealand, two introduced scolytid beetles, Hylastes ater and Hylurgus ligniperda (Curculionidae: Scolytinae) are pests in pine plantations. Investigation of the naturally occurring pathogens of these exotic pests revealed that both are attacked by Beauveria caledonica, a species originally isolated and described from soil in Scotland. The isolates in New Zealand were identical in morphology and conserved DNA region (rDNA, elongation factor alpha) sequence to isolates held in the USDA-ARS insect pathogens culture collection. In bioassay, the B. caledonica isolates were highly pathogenic to adults of H. ligniperda and larvae of Tenebrio molitor. Sporulation was observed on cadavers, confirming the species can utilise the cadavers. As both species were likely to have been introduced to New Zealand from Europe, a search was made for B. caledonica in the northern UK and Ireland. The fungus was found as a naturally-occurring pathogen of the weevil pest, Hylobius abietis (Curculionidae: Scolytinae), developing in spruce and other beetles in forests in both regions.

Virulence of nucleopolyhedroviruses from Orgyia pseudotsugata and Orgyia leucostigma (Lep., Lymantriidae) for early instars of the white-spotted tussock moth, Orgyia thyellina

The white‐spotted tussock moth, Orgyia thyellina, was found in Auckland, New Zealand, in 1995. An eradication campaign against this potential pest is currently underway, but in the event that it is unsuccessful, agents for population suppression will be required. As no pathogens were available from O. thyellina, two nucleopolyhedroviruses (NPVs) from Orgyia pseudotsugara (OpMNPV ‘VirtussTM’) and Orgyia leucostigma (OrleSNPV) were imported from Canada and tested for virulence against O. thyellina. At 105 polyhedral inclusion bodies per cm2 and above, 100% mortality was achieved with first to second instar larvae by 15 days. Molecular comparison of sequences from the pol gene region of each virus demonstrated that they were from different genotypic groups of NPVs.

Formation of microsclerotia in three species of Beauveria and storage stability of a prototype granular formulation

ABSTRACT Beauveria bassiana is the only species from the Beauveria genus that has been previously described as forming microsclerotia [Wang, H.-H., Wang, J.-L., Li, Y.-P., Liu, X., Wen, J.-Z., & Lei, Z.-R. (2011). Liquid culturing of microsclerotia of Beauveria bassiana, an entomopathogenic fungus to control western flower thrip, Frankliniella occidentalis in Journal of Applied Entomology, 48(3), 588–595]. Microsclerotia (MS) are compact, melanised aggregates that have been recognised in plant pathogenic fungi as overwintering structures. MS were successfully produced in shake flask cultures from three different Beauveria species (B. bassiana, B. brogniartii and B. pseudobassiana) using liquid media containing a low carbon to nitrogen ratio and replacing glucose with a complex carbon source. Maximum MS yield after 10 days fermentation was 8.46 × 103 MS/mL obtained with B. pseudobassiana AgR-F704, a New Zealand fungus isolated from the African black beetle (Heteronychus arator F.). Using this isolate, MS were produced using two C:N compositions, formulated with diatomaceous earth and air dried and monitored for germination and storage stability. Germination of MS granules varied with C:N ratio and storage temperature. MS produced using a higher carbon concentration maintained 100% germination after six months of storage at 4°C, but when stored at 20°C, the germination rate was reduced to 68%. In contrast, MS produced using a lower carbon concentration showed reduced germination (0 to 18%) after storage for more than two months at either temperature and had lost viability completely after six months. Conidia production from surviving MS was estimated at 109 conidia/g of granules. Results demonstrated that Beauveria species are able to form MS in submerged culture and those produced in the relatively higher carbon medium were capable of surviving under low moisture, ambient temperature storage and are able to produce conidia after germination; all of these attributes suggest that MS can be incorporated into granules as a new sustained release strategy to control soil-dwelling insects.