Production of bacteriocins by Streptococcus bovis strains from Australian ruminants.

Aims: To examine the prevalence of bacteriocin production in Streptococcus bovis isolates from Australian ruminants and the feasibility of industrial production of bacteriocin. Methods and Results: Streptococcus bovis strains were tested for production of bacteriocin-like inhibitory substances (BLIS) by antagonism assay against Lactococcus lactis. BLIS production was associated with source animal location (i.e. proximity of other bacteriocin-positive source animals) rather than ruminant species/breed or diet. One bacteriocin showing strong inhibitory activity (Sb15) was isolated and examined. Protein sequence, stability and activity spectrum of this bovicin were very similar to bovicin HC5. Production could be increased through serial culturing, and increased productivity could be partially maintained during cold storage of cultures. Conclusions: BLIS production is geographically widely distributed in Eastern Australia, and it appears that the bacteriocin+ trait is maintained in animals at the same location. The HC5-like bacteriocin, originally identified in North America, is also found in Australia. Production of bacteriocin can be increased through serial culturing. Significance and Impact of the Study: The HC5-like bacteriocins appear to have a broad global distribution. Serial culturing may provide a route towards commercial manufacturing for use in industrial applications, and purified bacteriocin from S. bovis Sb15 could potentially be used to prevent food spoilage or as a feed additive to promote growth in ruminant species.

Wood properties and knot occlusion of plantation grown Eucalyptus dunnii and Corymbia citriodora subsp. variegata in a pruning and thinning experiment.

A 2 × 2 factorial combination of thinned or unthinned, and pruned or unpruned 11-year-old Eucalyptus dunnii (DWG) and 12-year-old Corymbia citriodora subsp. variegata (CCV) was destructively sampled to provide 60 trees in total per species. Two 1.4 m long billets were cut from each tree and were rotary veneered in a spindleless lathe down to a 45 mm diameter core to expose knots which were classified as either alive, partially occluded or fully occluded. Non-destructive evaluation of a wider range of thinning treatments available in these trials was undertaken with Pilodyn and Fakopp tools. Disc samples were also taken for basic density and modulus of elasticity. Differences between treatments for all wood property assessments were generally small and not significantly different.Thinning and pruning had little effect on the stem diameter growth required to achieve occlusion, therefore occlusion would be more rapid after thinning due to more rapid stem diameter growth. The difference between the treatments of greatest management interest, thinned and pruned (T&P) and unthinned and unpruned (UT&UP) were small. The production of higher value clear wood produced after all knots had occluded, measured as the average stem diameter growth over occlusion of the three outermost knots, was approximately 2 centimetres diameter. Two of the treatments can be ruled out as viable management alternatives: (i) the effect of thinning without pruning (T&UP) is clear, leading to a large inner core of stem wood containing knots (large knotty core diameter) and (ii) pruning without thinning (UT&P) results in a small knotty core diameter, however the tree and therefore log diameters are also small.

Salmonella Virchow and Salmonella Weltevreden in a Random Survey of the Asian House Gecko, Hemidactylus frenatus, in Houses in Northern Australia.

Background: Salmonella enterica serotype Virchow is the most common cause of invasive nontyphoid salmonellosis in North Queensland, particularly in infants, but the zoonotic source is unknown. This study aimed at determining (i) the prevalence of the introduced Asian house gecko, Hemidactylus frenatus, in houses in North Queensland and (ii) whether they were a potential source of Salmonella Virchow. Methods: Asian house geckos were collected in a random survey of houses in Townsville, North Queensland. Gut contents underwent microbiological analysis within 2 h of removal using both direct plating and enrichment broth methods. Any organism found to be a presumptive Salmonella spp. was then sent to a reference lab for confirmation of genus/species, serotyping, and phage typing if indicated. Results: One hundred Asian house geckos were collected from 57 houses. Geckos were present in 100% of houses surveyed, and prevalence of Salmonella in large intestinal contents was 7% (95% confidence interval 2, 12%). Three serotypes were found: Virchow (phage type 8), Weltevreden, and an untypable subspecies 1 serotype 11:-:1,7. Conclusion: Since Salmonella Virchow (phage type 8) is associated with invasive disease, the introduced Asian house gecko may play a significant role in the epidemiology of sporadic salmonellosis in places invaded by these peridomestic reptiles. These results justify more detailed epidemiological studies on the role of the Asian house gecko in sporadic salmonellosis and development of evidence-based strategies to decrease this potential zoonotic hazard.

Gene expression evidence for off-target effects caused by RNA interference-mediated gene silencing of Ubiquitin-63E in the cattle tick Rhipicephalus microplus.

Knowledge of cattle tick (Rhipicephalus (Boophilus) microplus; Acari: Ixodidae) molecular and cellular pathways has been hampered by the lack of an annotated genome. In addition, most of the tick expressed sequence tags (ESTs) available to date consist of similar to 50% unassigned sequences without predicted functions. The most common approach to address this has been the application of RNA interference (RNAi) methods to investigate genes and their pathways. This approach has been widely adopted in tick research despite minimal knowledge of the tick RNAi pathway and double-stranded RNA (dsRNA) uptake mechanisms. A strong knockdown phenotype of adult female ticks had previously been observed using a 594 bp dsRNA targeting the cattle tick homologue for the Drosophila Ubiquitin-63E gene leading to nil or deformed eggs. A NimbleGen cattle tick custom microarray based on the BmiGI.V2 database of R. microplus ESTs was used to evaluate the expression of mRNAs harvested from ticks treated with the tick Ubiquitin-63E 594 bp dsRNA compared with controls. A total of 144 ESTs including TC6372 (Ubiquitin-63E) were down-regulated with 136 ESTs up-regulated following treatment. The results obtained substantiated the knockdown phenotype with ESTs identified as being associated with ubiquitin proteolysis as well as oogenesis, embryogenesis, fatty acid synthesis and stress responses. A bioinformatics analysis was undertaken to predict off-target effects (OTE) resulting from the in silico dicing of the 594 bp Ubiquitin-63E dsRNA which identified 10 down-regulated ESTs (including TC6372) within the list of differentially expressed probes on the microarrays. Subsequent knockdown experiments utilising 196 and 109 bp dsRNAs, and a cocktail of short hairpin RNAs (shRNA) targeting Ubiquitin-63E, demonstrated similar phenotypes for the dsRNAs but nil effect following shRNA treatment. Quantitative reverse transcriptase PCR analysis confirmed differential expression of TC6372 and selected ESTs. Our study demonstrated the minimisation of predicted OTEs in the shorter dsRNA treatments (similar to 100-200 bp) and the usefulness of microarrays to study knockdown phenotypes.

Investigation of potential antibacterial action for postharvest copper treatments of cut Acacia holosericea

The mechanisms of action of Cu 2+ in improving the longevity of cut flowers and foliage have not been elucidated. Possible antimicrobial action of Cu 2+ against stem end and vase solution colonising bacteria was investigated using Cu 2+ treatments optimised for cut Acacia holosericea A. Cunn. ex G. Don foliage stems. These treatments were a 5h pulse with 2.2mM Cu 2+ or a 0.5mM Cu 2+ vase solution versus a deionised water (no Cu 2+) control. Bacterial growth over time was assessed by a standard plate count agar technique and with scanning electron microscopy. Cu 2+ treatments significantly extended the cut foliage vase life. However, they did not have sustained antibacterial activity against stem end or vase solution colonising bacteria. Also, regular recutting of 1-2cm from the stem ends did not substantially improve either cut stem water relations or longevity. The positive effects of Cu 2+ treatments were unaffected by the repeated stem end recutting. It was concluded that the primary mechanism of Cu 2+ was not antibacterial. Moreover, naturally growing vase solution and stem end microbial populations had relatively insignificant effects on cut A. holosericea vase life. Research into alternative mechanisms of Cu 2+ is required. © 2012 Elsevier B.V.

Biocontrol of Chromolaena odorata in Papua New Guinea

Chromolaena odorata (L.) King and Robinson (Asteraceae) is a significant agricultural weed in Papua New Guinea (PNG), affecting plantations, food gardens and grazing lands. It was the focus of a collaborative biocontrol program funded by the Australian Government between 1998 and 2007. Chromolaena was recorded at 680 sites in 13 provinces of PNG through surveys, field releases of biocontrol agents and feedback from public awareness programs. Three biocontrol agents, the moth Pareuchaetes pseudoinsulata Rego Barros (Lepidoptera: Arctiidae), the stemgalling fly Cecidochares connexa (Macquart) (Diptera: Tephritidae) and the leaf mining fly Calycomyza eupatorivora Spencer (Diptera: Agromyzidae), were introduced to control chromolaena. Cecidochares connexa was found to be the most effective of the agents introduced as it quickly established at over 300 sites where it was released and spread up to 100km in five years from some sites. Experimental field plots established to determine the impact of the agents on chromolaena, showed that the size of chromolaena infestations decreased with the presence of C. connexa. A survey was conducted to quantify the social and economic benefits of biocontrol of chromolaena to landholders. Chromolaena is considered to be under substantial/significant control in nine provinces in PNG, with about 50% of respondents stating that there is less than 50% of chromolaena remaining following the release of the gall fly. This has resulted in landholders spending less time clearing chromolaena and the re-establishment of small-scale subsistence farms and the regeneration of natural vegetation. Crop yield and income generated from the sale of agricultural produce have increased by at least 50% since chromolaena was brought under biocontrol. It is anticipated that the gall fly will continue to spread and control chromolaena in areas where it has not yet reached, thereby further reducing the impact of the weed in PNG.

Estimating an Aggregate Economic Genotype to Facilitate Breeding and Selection of Strawberry Genotypes in Southeast Queensland

The Queensland strawberry (Fragaria ×ananassa) breeding program in subtropical Australia aims to improve sustainable profitability for the producer. Selection must account for the relative economic importance of each trait and the genetic architecture underlying these traits in the breeding population. Our study used estimates of the influence of a trait on production costs and profitability to develop a profitability index (PI) and an economic weight (i.e., change in PI for a unit change in level of trait) for each trait. The economic weights were then combined with the breeding values for 12 plant and fruit traits on over 3000 genotypes that were represented in either the current breeding population or as progenitors in the pedigree of these individuals. The resulting linear combination (i.e., sum of economic weight × breeding value for all 12 traits) estimated the overall economic worth of each genotype as H, the aggregate economic genotype. H values were validated by comparisons among commercial cultivars and were also compared with the estimated gross margins. When the H value of ‘Festival’ was set as zero, the H values of genotypes in the pedigree ranged from –0.36 to +0.28. H was highly correlated (R2 = 0.77) with the year of selection (1945–98). The gross margins were highly linearly related (R2 > 0.98) to H values when the genotype was planted on less than 50% of available area, but the relationship was non-linear [quadratic with a maximum (R2 > 0.96)] when the planted area exceeded 50%. Additionally, with H values above zero, the variation in gross margin increased with increasing H values as the percentage of area planted to a genotype increased. High correlations among some traits allowed the omission of any one of three of the 12 traits with little or no effect on ranking (Spearman’s rank correlation 0.98 or greater). Thus, these traits may be dropped from the aggregate economic genotype, leading to either cost reductions in the breeding program or increased selection intensities for the same resources. H was efficient in identifying economically superior genotypes for breeding and deployment, but because of the non-linear relationship with gross margin, calculation of a gross margin for genotypes with high H is also necessary when cultivars are deployed across more than 50% of the available area.

Field and laboratory evaluation of fungicides for the control of Phytophthora fruit rot of papaya in far north Queensland, Australia

Results from the first of two artificially inoculated field experiments showed foliar applications of copper hydroxide (Blue Shield Copper) at 600 g a.i./100 L−1 (0% infected fruit), copper hydroxide + metalaxyl-M (Ridomil Gold Plus.) at 877.5 g a.i./100 L−1 (0.27%), metiram + pyraclostrobin (Aero) at 720 g a.i./100 L−1 (0.51%), chlorothalonil (Bravo WeatherStik) at 994 g a.i./100 L−1 (0.63%) and cuprous oxide (Nordox 750 WG) at 990 g a.i./100 L−1 (0.8%) of water significantly reduced the percentage of infected fruit compared to potassium phosphonate (Agri-Fos 600) at 1200 g a.i./100 L−1 (8.22%), dimethomorph (Acrobat) at 108 g a.i./100 L−1 (11.18%) and the untreated control (16%). Results from the second experiment showed fruit sprayed with copper hydroxide (Champ Dry Prill) at 300 (2.0% infected fruit), 375 (0.4%) and 450 g a.i./100 L−1 (0.6%) and metiram + pyraclostrobin (Aero) at 360 (2.8%), 480 (0.6%) and 600 g a.i./100 L−1 of water (1.0%) significantly reduced the percentage of infected fruit compared to the untreated control (19.4%). Foliar sprays of copper hydroxide at 375 g a.i./100 L−1 in rotation with chlorothalonil at 994 g a.i./100 L−1 every two weeks is now recommended to growers for controlling Phytophthora fruit rot of papaya.

Longitudinal cohort study of horse owners

This report summarises the findings of a three-year mixed methods research study designed to capture factors that influence horse owner Hendra virus (HeV) risk mitigation practices. The research project focuses on horse owners; their knowledge, attitudes, and risk mitigation practices, i.e. uptake of vaccination, property management, and biosecurity practices. A flexible research methodology enabled the tracking of core subject areas over time whilst also responding to new or evolving shifts in the HeV landscape, e.g. new HeV cases, event management, and issues arising in the vaccine roll-out. By tracking relationships within the data and engaging with stakeholders and the horse owner population, it is hoped that findings from the study will help to identify important linkages and effective strategies for communication/information and policy implementation.