Development of a multi-locus sequence typing scheme for avian isolates of Pasteurella multocida.

A total of 63 isolates of Pasteurella multocida from Australian poultry, all associated with fowl cholera outbreaks, and three international reference strains, representing the three subspecies within P. multocida were used to develop a multi-locus sequence typing scheme. Primers were designed for conserved regions of seven house-keeping enzymes - adk, est, gdh, mdh, pgi, pmi and zwf - and internal fragments of 570-784 bp were sequenced for all isolates and strains. The number of alleles at the different loci ranged from 11 to 20 and a total of 29 allelic profiles or sequence types were recognised amongst the 66 strains. There was a strong concordance between the MLST data and the existing multi-locus enzyme electrophoresis and ribotyping data. When used to study a sub-set of isolates with a known detailed epidemiological history, the MLST data matched the results given by restriction endonuclease analysis, pulsed-field gel electrophoresis, ribotyping and REP-PCR. The MLST scheme provides a high level of resolution and is an excellent tool for studying the population structure and epidemiology of P. multocida.

Inheritance and Characterization of Strong Resistance to Phosphine in Sitophilus oryzae(L.)

Sitophilus oryzae (Linnaeus) is a major pest of stored grain across Southeast Asia and is of increasing concern in other regions due to the advent of strong resistance to phosphine, the fumigant used to protect stored grain from pest insects. We investigated the inheritance of genes controlling resistance to phosphine in a strongly resistant S. oryzae strain (NNSO7525) collected in Australia and find that the trait is autosomally inherited and incompletely recessive with a degree of dominance of -0.66. The strongly resistant strain has an LC50 52 times greater than a susceptible reference strain (LS2) and 9 times greater than a weakly resistant strain (QSO335). Analysis of F2 and backcross progeny indicates that two or more genes are responsible for strong resistance, and that one of these genes, designated Sorph1, not only contributes to strong resistance, but is also responsible for the weak resistance phenotype of strain QSO335. These results demonstrate that the genetic mechanism of phosphine resistance in Soryzae is similar to that of other stored product insect pests. A unique observation is that a subset of the progeny of an F1 backcross generation are more strongly resistant to phosphine than the parental strongly resistant strain, which may be caused by multiple alleles of one of the resistance genes.

Cross-protection study of the nine serovars of Haemophilus paragallinarum in the Kume haemagglutinin scheme

The cross-protection and haemagglutination-inhibition antibodies present in chickens vaccinated with one of the nine currently recognized Kume haemagglutinin serovars of Haemophilus paragallinarum were investigated. The results confirmed the widely accepted dogma that serogroups A, B, and C represent three distinct immunovars. Within Kume serogroup A, there was generally good cross-protection among all four serovars. However, within Kume serogroup C, there was evidence of a reduced level of cross protection between some of the four serovars. The haemagglutination-inhibition antibody levels generally showed the same trend as with the cross-protection results. This study suggests that some apparent field failures of infectious coryza vaccines may be due to a lack of cross-protection between the vaccine strains and the field strains. Our results will help guide the selection of strains for inclusion in infectious coryza vaccines.

Baseline responses of Alphitobius diaperinus (Coleoptera : Tenebrionidae) to cyfluthrin and detection of strong resistance in field populations in eastern Australia

Resistance to cyfluthrin in broiler farm populations of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in eastern Australia was suspected to have contributed to recent control failures. In 2000-2001, beetles from 11 broiler farms were tested for resistance by comparing them to an insecticide-susceptible reference population by using topical application. Resistance was detected in almost all beetle populations (up to 22 times the susceptible at the LC50), especially in southeastern Queensland where more cyfluthrin applications had been made. Two from outside southeastern Queensland were found to be susceptible. Dose-mortality data generated from the reference population over a range of cyflutbrin concentrations showed that 0.0007% cyfluthrin at a LC99.9 level could be used as a convenient dose to discriminate between susceptible and resistant populations. Using this discriminating concentration, from 2001 to 2005, the susceptibilities of 18 field populations were determined. Of these, 11 did not exhibit complete mortality at the discriminating concentration (mortality range 2.8-97.7%), and in general, cyfluthrin resistance was directly related to the numbers of cyfluthrin applications. As in the full study, populations outside of southeastern Queensland were found to have lower levels of resistance or were susceptible. One population from an intensively farmed broiler area in southeastern Queensland exhibited low mortality despite having no known exposure to cyfluthrin. Comparisons of LC50 values of three broiler populations and a susceptible population, collected in 2000 and 2001 and recollected in 2004 and 2005 indicated that values from the three broiler populations had increased over this time for all populations. The continued use of cyfluthrin for control of A. diaperinus in eastern Australia is currently under consideration.

Detection and characterisation of strong resistance to phosphine in Brazilian Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae)

As failure to control Rhyzopertha dominica (F.) with phosphine is a common problem in the grain-growing regions of Brazil, a study was undertaken to investigate the frequency, distribution and strength of phosphine resistance in R. dominica in Brazil. Nineteen samples of R. dominica were collected between 1991 and 2003 from central storages where phosphine fumigation had failed to control this species. Insects were cultured without selection until testing in 2005. Each sample was tested for resistance to phosphine on the basis of the response of adults to discriminating concentrations of phosphine (20 and 48 h exposures) and full dose-response assays (48 h exposure). Responses of the Brazilian R. dominica samples were compared with reference susceptible, weak-resistance and strong-resistance strains from Australia in parallel assays. All Brazilian population samples showed resistance to phosphine: five were diagnosed with weak resistance and 14 with strong resistance. Five samples showed levels of resistance similar to the reference strong-resistance strain. A representative highly resistant sample was characterised by exposing mixed-age cultures to a range of constant concentrations of phosphine for various exposure periods. Time to population extinction (TPE) and time to 99.9% suppression of population (LT99.9) values of this sample were generally similar to those of the reference strong-resistance strain. For example, at 0.1, 0.5 and 1.0 mg L-1, LT99.9 values for BR33 and the reference strong-resistance strain were respectively 21, 6.4 and 3.7 days and 17, 6.2 and 3.8 days. With both strains, doubling phosphine concentrations to 2 mg L -1 resulted in increased LT99.9 and TPE. High level and frequency of resistance in all population samples, some of which had been cultured without selection for up to 12 years, suggest little or no fitness deficit associated with phosphine resistance. The present research indicates that widespread phosphine resistance may be developing in Brazil. Fumigation practices should be monitored and resistance management plans implemented to alleviate further resistance development.

Action learning in partnership with Landcare and catchment management groups to support increased pasture sowings in southern inland Queensland

The incorporation of sown pastures as short-term rotations into the cropping systems of northern Australia has been slow. The inherent chemical fertility and physical stability of the predominant vertisol soils across the region enabled farmers to grow crops for decades without nitrogen fertiliser, and precluded the evolution of a crop–pasture rotation culture. However, as less fertile and less physically stable soils were cropped for extended periods, farmers began to use contemporary farming and sown pasture technologies to rebuild and maintain their soils. This has typically involved sowing long-term grass and grass–legume pastures on the more marginal cropping soils of the region. In partnership with the catchment management authority, the Queensland Murray–Darling Committee (QMDC) and Landcare, a pasture extension process using the LeyGrain™ package was implemented in 2006 within two Grain & Graze projects in the Maranoa-Balonne and Border Rivers catchments in southern inland Queensland. The specific objectives were to increase the area sown to high quality pasture and to gain production and environmental benefits (particularly groundcover) through improving the skills of producers in pasture species selection, their understanding and management of risk during pasture establishment, and in managing pastures and the feed base better. The catalyst for increasing pasture sowings was a QMDC subsidy scheme for increasing groundcover on old cropping land. In recognising a need to enhance pasture knowledge and skills to implement this scheme, the QMDC and Landcare producer groups sought the involvement of, and set specific targets for, the LeyGrain workshop process. This is a highly interactive action learning process that built on the existing knowledge and skills of the producers. Thirty-four workshops were held with more than 200 producers in 26 existing groups and with private agronomists. An evaluation process assessed the impact of the workshops on the learning and skill development by participants, their commitment to practice change, and their future intent to sow pastures. The results across both project catchments were highly correlated. There was strong agreement by producers (>90%) that the workshops had improved knowledge and skills regarding the adaptation of pasture species to soils and climates, enabling a better selection at the paddock level. Additional strong impacts were in changing the attitudes of producers to all aspects of pasture establishment, and the relative species composition of mixtures. Producers made a strong commitment to practice change, particularly in managing pasture as a specialist crop at establishment to minimise risk, and in the better selection and management of improved pasture species (particularly legumes and the use of fertiliser). Producers have made a commitment to increase pasture sowings by 80% in the next 5 years, with fourteen producers in one group alone having committed to sow an additional 4893 ha of pasture in 2007–08 under the QMDC subsidy scheme. The success of the project was attributed to the partnership between QMDC and Landcare groups who set individual workshop targets with LeyGrain presenters, the interactive engagement processes within the workshops themselves, and the follow-up provided by the LeyGrain team for on-farm activities.

Cynodon dactylon, Green Couch Grass 'TL1'

Chance seedling: observed in about 1989 as a distinctly coarser textured, densely matting, darker green mutant bermuda grass plant growing among the hybrid ‘Tifgreen’ on the eighth green at the Townsville Golf Course. Although ‘TL1’ was selected from a sward of the hybrid Bermuda grass ‘Tifgreen’, its inflorescence structure (4, not 3, racemes per inflorescence), agronomic attributes (e.g. its tolerance to certain herbicides), and its DNA profile are consistent with a chance seedling of Cynodon dactylon rather than a mutant plant of hybrid (C. dactylon x transvaalensis) origin. Selection criteria: exceptionally short stolon internodes resulting in an extremely tight knit stolon mat under close (c. 5-6 mm) but not very close (c. 3-4 mm) mowing; very deep, strong rhizome system; very dark green colour; tolerates shade better than other Australian bermuda grass varieties of common knowledge (except for ‘Plateau’A); and remains low growing under heavy tropical cloud cover even after 6-8 months. Designated ‘TL1’ by Tropical Lawns Pty Ltd and trialed successfully during the late 1990s and early 2000s in high wear situations (e.g. golf tees) in north Queensland. Propagation: vegetative. Breeder: Barry McDonagh, Townsville, QLD. PBR Certificate Number 2638, Application Number 2002/267, granted 24 February 2005.

Sorghum Midge Tested Scheme

Scheme provides independant testing and official industry midge resistance rating of pre-commercial sorghum varieities. Continuation of current project.

Sorghum Midge Testing Scheme

Scheme provides independant testing and official industry midge resistance rating of pre-commercial sorghum varieities.

Short-range resource location by Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) demonstrates a strong preference for fungi associated with cotton seed

Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is a common stored grain pest for which a wide range of suitable resources has been recorded. These beetles are facultatively fungivorous and their resource range may extend to fungi associated with non-grain resources (e.g. cotton seed) and other decaying plant matter. Little is known with respect to fungi in terms of resource location by these beetles in the field. We, therefore, conducted a series of experiments in laboratory arenas, glasshouse cages and the field to determine how beetles respond to grain resources in relation to cotton seed (together with its lint stubble and associated fungal flora). Results from the tests conducted in relatively small arenas and cages in the laboratory and glasshouse reveal that the responses of T. castaneum adults to food resources were twice as strong when walking as when flying (as measured by the proportion of the released beetles that were trapped). Also, a clear preference for linted cotton seeds was evident in walking T. castaneum, especially in small-scale arenas in the laboratory, where at least 60% of beetles released preferred linted cotton seeds over wheat and sorghum. Similarly, in cages (1 m3) they responded five times more strongly to linted cotton seed than to conventional grain resources. However, this pattern was not consistent with those obtained from field trapping over 20 m and the beetles did not show any particular preference to any of the resources tested above. Our results suggest a focus on walking beetles in trapping studies for population estimations and, for developing effective food-based trapping lures, the potential use of active volatiles from the fungi associated with linted cotton seed. © 2012 Elsevier Ltd.

Strong resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae): its characterisation, a rapid assay for diagnosis and its distribution in Australia

BACKGROUND: The recent development of very high resistance to phosphine in rusty grain beetle, Cryptolestes ferrugineus (Stephens), seriously threatens stored-grain biosecurity. The aim was to characterise this resistance, to develop a rapid bioassay for its diagnosis to support pest management and to document the distribution of resistance in Australia in 20072011. RESULTS: Bioassays of purified laboratory reference strains and field-collected samples revealed three phenotypes: susceptible, weakly resistant and strongly resistant. With resistance factors of > 1000 x , resistance to phosphine expressed by the strong resistance phenotype was higher than reported for any stored-product insect species. The new time-to-knockdown assay rapidly and accurately diagnosed each resistance phenotype within 6 h. Although less frequent in western Australia, weak resistance was detected throughout all grain production regions. Strong resistance occurred predominantly in central storages in eastern Australia. CONCLUSION: Resistance to phosphine in the rusty grain beetle is expressed through two identifiable phenotypes: weak and strong. Strong resistance requires urgent changes to current fumigation dosages. The development of a rapid assay for diagnosis of resistance enables the provision of same-day advice to expedite resistance management decisions. (c) 2012 Commonwealth of Australia. Published by John Wiley & Sons, Ltd.