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Coconut production in the Pacific Islands is threatened by coconut rhinoceros beetle (CRB). Distinctive feeding damage caused by adults is often the first indicator of CRB activity in a new location. Multiple approaches to damage assessment have been used, reflecting observers' experience. The lack of standardisation makes it difficult to compare CRB impacts on coconut palms over time. We examine damage assessment from three perspectives: participants, power and purpose. Participants applied binary, 3-point and 5-point damage scales consecutively to photographs of damaged palms. Participant responses were more accurate using the binary scale compared with the 5-point scale. Experience with CRB improved accuracy, but this did not wholly compensate for the effect of scale complexity. Calculations of statistical power determined minimum sample sizes to detect a 20% change in palm damage was 50 palms for the binary scale, 150 palms for the 3-point scale and > 200 palms for the 5-point scale, when the change in damage was spread across damage categories. More complex damage scales did not increase the likelihood of detecting significant changes in palm damage. With real-world examples, including detecting new outbreaks and managing established populations, recommendations were developed to standardise damage assessment for CRB on coconut palms. [ABSTRACT FROM AUTHOR]

Beauveria pseudobassiana formed three-dimensional aggregates of cells (CAs) in liquid culture. CAs were formed mainly by blastospores and conidia, distinct from microsclerotia formed through adhesion of hyphae. The formation, germination and sporulation of CAs were studied, as well as the pathogenicity of conidia produced from them against adults of black beetle. After 4 days of culture, CAs were formed, becoming compact and melanised after 10 days of incubation. Electron microscopy showed three-dimensional CAs averaging 431.65 µm in length with irregular shapes and rough surfaces, where cells were trapped within an extracellular matrix. CAs germinated after 2 days of incubation on agar-plates producing hyphae and forming phialides and conidia after 4 days. Produced conidia caused 45% mortality of black beetle adults. CAs germination and sporulation on soil were directly correlated with soil moisture, reaching 80% and 100% germination on the surface of soil with 17% and 30% moisture, respectively. CAs maintained 100% germination after 2 years of storage under refrigeration. These CAs could have a similar function as microsclerotia in nature, acting as resistant structures able to protect internal cells and their ability to sporulate producing infective conidia, suggesting their potential to be used as bioinsecticides to control soil-dwelling insects. [ABSTRACT FROM AUTHOR]

Some entomopathogenic fungi such as Metarhizium and Beauveria not only have the ability to infect and kill insects but also the capability to associate with plant roots. The potential benefits from this plant-fungus association include nutrient acquisition, plant growth promotion and protection against stressors. The objective of this study was to evaluate the delivery of entomopathogenic fungi in seed coating to determine whether the fungal presence affected maize performance. Additionally, fungal biocontrol ability was assessed in terms of plant resistance to the larvae of Costelytra giveni (Coleoptera: Scarabaeidae) and to the fungus Fusarium graminearum (Nectriaceae). Maize seeds were coated with conidia from Metarhizium spp. or Beauveria bassiana and plant performance was evaluated as seed germination and plant dry weight. Larval mortality and the presence of Fusarium root rot symptoms were also determined. The entomopathogenic fungal persistence on the maize rhizosphere was demonstrated with a M. anisopliae isolate expressing the green fluorescent protein. Presence of both challengers was detrimental to maize performance with 33% reduction in root dry weight in control plants while no variation was observed when the entomopathogenic fungi were used to coat seeds. Some seed coatings resulted in up to 67% mycosis of C. giveni larvae and a reduction in Fusarium rot root symptoms between 24 – 44%. This study showed that seed coating with conidia of Metarhizium or Beauveria can be used as a delivery system for pests and plant pathogen control, while at the same time hyphae formed a close association with plant roots after conidial germination. [ABSTRACT FROM AUTHOR]

The invertebrate pests most commonly affecting New Zealand's pastoral-based production in 'average' years cause losses of between $1.7B and $2.3B p.a. of which up to $0.9B occur on sheep and beef farms and $1.4B on dairy farms. The native scarab grass grub is the most costly pest causing losses of $140-380 M on dairy farms and $75-205 M on sheep and beef farms annually. The exotic scarab, black beetle, although only affecting approximately 1 M ha, costs dairy farmers up to $223 M and sheep and beef farmers up to $19 M annually. Porina cause losses up to $84 M and $88 M respectively. Pasture nematodes are estimated to cost up to $274 M p.a. for dairy farmers and $326 M p.a. for sheep and beef farmers. Two exotic pests, Argentine stem weevil (ASW) and clover root weevil (CRW) are causing damage estimated at up to $200 M p.a. and $235 M p.a. respectively in dairy and sheep and beef pastures. While CRW is subject to successful biological control management it still causes considerable losses. Lesser pests also contribute to lost production, particularly as they often coexist with more major pests. However, their economic cost to New Zealand is difficult to calculate due to the variable nature of infestations on both temporal and spatial scales. At farm and paddock level, it is abundantly clear that substantial savings could be made if pest management is achieved. It is equally clear that in many instances the tools to do so are limited but if developed would contribute substantially to farm profitability. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The nonspore-forming bacteriumSerratia entomophilamay be used to control the New Zealand grass grub (Costelytra giveni) but is sensitive to environmental stress and must be formulated to improve survival. Existing formulations require subsurface application limiting the area that can be treated. Formulations that allow delivery by broadcast methods are desirable to reduce application costs and increase the potential for aerial application to inaccessible areas. Two formulations were prepared for use in experiments examining the persistence and movement of inoculum through soil. When granules were applied to the soil surface, bacterial survival was negligible in uncoated core, but improved with increasing thickness of the coating. Both survival of bacteria and release into the soil were influenced by soil moisture content. Granules at <12% soil moisture showed high bacterial mortality and reduced delivery to the soil, while at 28% soil moisture most bacteria were released to the soil. There was a high level of survival of the applied bacteria within granules at 20% and 28% soil moisture. The formulations maintained viability ofS. entomophilain granules stored under ambient conditions for more than 6 months. In laboratory and field tests, the application of granules caused disease in the target grass grub larvae, whether application was applied to the surface or subsurface. In field trials, broadcast applied granules could produce equivalent disease to thin-coat granules drilled into the soil, but these levels of disease were associated with the occurrence of precipitation shortly after application. [ABSTRACT FROM AUTHOR]

The bacterium Serratia entomophila (Enterobacteriaceae) has been developed as a commercially available biopesticide for control of the pasture pest Costelytra zealandica. The influence of culture medium composition, dissolved oxygen (DO) concentration and harvesting time were investigated in order to optimise the production of S. entomophila. In batch fermentations, highest yields were achieved using sucrose (40 g L-1) as the carbon source, followed closely by fructose and molasses. The effect of yeast extract (YE), marmite and bakery yeast as cell growth enhancers was also examined in both batch and fed-batch mode. Culture medium containing 20 g L-1 of YE (fed-batch) produced the highest cell density. No significant effect on cell yield was detected when cultures were supplemented with bakery yeast or marmite. The DO concentration influenced biomass production: a 5-fold increase in cell density was achieved when the concentration of DO was maintained in the range of 20-50% (5.7×1010 CFUs mL-1) in comparison with 1% (1.2×1010 CFUs mL-1). In cultures maintained at 1 and 20% DO concentration, cells harvested from the exponential growth phase survived for less than 2 weeks when stored at 4°C. In contrast, high cell survival (85-100%) was achieved when cells were harvested after they had entered the stationary growth phase. Recommendations are provided for the production of robust, high cell density cultures of S. entomophila. [ABSTRACT FROM AUTHOR]

Bacteria of the genus Serratia are often associated with insects, often carry distinctive pigments and can be readily isolated onto diagnostic selective media. Bacterial marking of parasitoids has been used in studies to determine rate of oviposition and host acceptance, and it provides a rapid indication of ovipositor penetration. The technique has been applied to parasitoid-weevil and parasitoid-lepidoptera interactions. Bacterial marking has the potential to be used in parasitoid dispersal studies and its use in IPM is discussed. [ABSTRACT FROM AUTHOR]

The Melolonthinae branch of the beetle family Scarabaeidae has evolved in isolation in Aotearoa, radiating into >100 endemic species, since Aotearoa separated from Gondwanaland 82 million years ago. The group includes important pasture pests, such as the New Zealand grass grub Costelytra giveni and the manuka beetle Pyronota festiva. These beetles, like other organisms, host their own distinctive microflora including beneficial microbial symbionts and pathogens. A wide range of microbial pathogens infect the Scarabaeidae, but in Aotearoa the bacteria Serratia entomophila, S. proteamaculans and S. quinivorans (Enterobacteriaceae) are frequently found causing natural disease epizootics in C. giveni. S. entomophila is widespread in Aotearoa pasture soils, with only rare isolations of S. entomophila documented in other countries. In contrast S. proteamaculans and S. quinivorans are globally ubiquitous, and are widely distributed within Aotearoa, with some isolates active against either C. giveni or Pyronota spp. larvae, or both. Virulence determinants that impart differential host specificity and potency are located on variants of the amber disease associated plasmid (pADAP). The host specificity of the Serratia-scarab system and the absence of similar systems in other geographies, suggests that the relationship between Serratia spp. and endemic scarabs has evolved in Aotearoa. [ABSTRACT FROM AUTHOR]