Updating the Quarantine Status of Prunus Infecting Viruses in Australia.

One hundred Prunus trees, including almond (P. dulcis), apricot (P. armeniaca), nectarine (P. persica var. nucipersica), peach (P. persica), plum (P. domestica), purple leaf plum (P. cerasifera) and sweet cherry (P. avium), were selected from growing regions Australia-wide and tested for the presence of 34 viruses and three viroids using species-specific reverse transcription-polymerase chain reaction (RT-PCR) or polymerase chain reaction (PCR) tests. In addition, the samples were tested using some virus family or genus-based RT-PCR tests. The following viruses were detected: Apple chlorotic leaf spot virus (ACLSV) (13/100), Apple mosaic virus (ApMV) (1/100), Cherry green ring mottle virus (CGRMV) (4/100), Cherry necrotic rusty mottle virus (CNRMV) (2/100), Cherry virus A (CVA) (14/100), Little cherry virus 2 (LChV2) (3/100), Plum bark necrosis stem pitting associated virus (PBNSPaV) (4/100), Prune dwarf virus (PDV) (3/100), Prunus necrotic ringspot virus (PNRSV) (52/100), Hop stunt viroid (HSVd) (9/100) and Peach latent mosaic viroid (PLMVd) (6/100). The results showed that PNRSV is widespread in Prunus trees in Australia. Metagenomic high-throughput sequencing (HTS) and bioinformatics analysis were used to characterise the genomes of some viruses that were detected by RT-PCR tests and Apricot latent virus (ApLV), Apricot vein clearing associated virus (AVCaV), Asian Prunus Virus 2 (APV2) and Nectarine stem pitting-associated virus (NSPaV) were also detected. This is the first report of ApLV, APV2, CGRMV, CNRNV, LChV1, LChV2, NSPaV and PBNSPaV occurring in Australia. It is also the first report of ASGV infecting Prunus species in Australia, although it is known to infect other plant species including pome fruit and citrus.

Effect of abattoir and cut on variations in microbial communities of vacuum-packaged beef.

This report builds on the earlier studies of the shelf-life of chilled Australian vacuum packaged (VP) beef primals (striploin and cube roll), products distinguished in the global marketplace for unusually long shelf-life. Notable findings in those studies were a shelf-life of at least 26weeks at -0.5°C, low microbial counts, and relatively high sensory scores. However, growth rates for total viable counts (TVC) and lactic acid bacteria (LAB) varied among the different abattoirs. The present study adds to these findings, by providing greater definition about temporal changes in bacterial communities using terminal restriction fragment length polymorphism (TRFLP) and clone library analyses of 16S ribosomal RNA (16S rRNA) gene, and measuring statistical associations among abattoir, beef cut, storage time and sensory attributes. Bacterial communities changed over time, with Carnobacterium spp. typically predominating (29-97%) at the end of storage. Variation in TRFLP profiles showed that different Carnobacterium strains predominated in different abattoirs, and that additional variation was due to the presence of other taxa typical of VP meat microbiomes. TRFLP-based community structure correlated significantly (P≤0.01) with sensorial characteristics, such as vacuum integrity, confinement odour, and intact pack appearance of beef. This study shows that Carnobacterium spp. predominate on extended shelf-life VP beef primals, while other taxa may produce subtle effects on shelf-life duration.

Multiplex RT-PCR detection of three common viruses infecting orchids.

A multiplex reverse transcription polymerase chain reaction (RT-PCR) assay was developed for simultaneous detection of three orchid viruses: cymbidium mosaic virus (CymMV), odontoglossum ringspot virus (ORSV), and orchid fleck virus (OFV). Primers were used to amplify nucleocapsid protein gene fragments of 845 bp (ORSV), 505 bp (CymMV) and 160 bp (OFV). A 60-bp amplicon of plant glyceraldehyde-3-phophate dehydrogenase mRNA was included as an internal control against false negatives. The assay was validated against 31 collected plants from six orchid genera and compared with results obtained by transmission electron microscopy (TEM). The RT-PCR assay proved more sensitive than TEM for detection of OFV.

Microbial growth, communities and sensory characteristics of vacuum and modified atmosphere packaged lamb shoulders.

Packaging fresh lamb in a vacuum (VAC) versus a 100% CO2 modified atmosphere (MAP) may influence product shelf-life and the bacterial communities. While VAC is a common packing method and 100% CO2 MAP is used in some countries, there is little information about how these different techniques affect the growth of spoilage bacteria and sensory attributes of lamb. The aim of this study was to assess changes in microbiological and organoleptic properties, and determine differences in microbial communities by terminal restriction fragment length polymorphism (TRFLP) and 454 pyrosequencing, in bone-in (BI) and bone-out (BO) MAP- and VAC-packed lamb shoulders stored at -0.3 °C over 12 wk. VAC and MAP lamb shoulders were acceptable in sensory test scores over 12 wk of storage at -0.3 °C, despite total viable count (TVC) and lactic acid bacteria (LAB) levels increasing to 8 log10 CFU/cm(2) for VAC lamb and 4-6 log10 CFU/cm(2) for MAP lamb. Similar to the sensory results, there were no significant differences in microbial communities between BI and BO product. However, types of bacteria were different between VAC and MAP packaging. Specifically, while VAC shoulder became dominated by Carnobacterium spp. in the middle of the storage period, the MAP shoulder microbial population remained similar from the start until later storage times.

Comparative assessment of genetic and epigenetic variation among regenerants of potato (Solanum tuberosum) derived from long-term nodal tissue-culture and cell selection.

Three long-term nodal tissued cultured Russet Burbank potato clones and nine thaxtomin A-treated regenerant lines, derived from the nodal lines, were assessed for genetic and epigenetic (in the form of DNA methylation) differences by AFLP and MSAP. The treated regenerant lines were originally selected for superior resistance to common scab disease and acceptable tuber yield in pot and field trials. The long-term, tissue culture clone lines exhibited genetic (8.75-15.63% polymorphisms) and epigenetic (12.56-26.13% polymorphisms) differences between them and may represent a stress response induced by normal plant growth disruption. The thaxtomin A-treated regenerant lines exhibited much higher significant (p < 0.05) genetic (2-29.38%) and epigenetic (45.22-51.76%) polymorphisms than the nodal cultured parent clones. Methylation-sensitive mutations accumulated within the regenerant lines are significantly correlated (p < 0.05) to disease resistance. However, linking phenotypic differences that could be of benefit to potato growers, to single gene sequence polymorphisms in a tetraploid plant such as the potato would be extremely difficult since it is assumed many desirable traits are under polygenic control.

Systematic prediction error correction: a novel strategy for maintaining the predictive abilities of multivariate calibration models.

The development of reliable multivariate calibration models for spectroscopic instruments in on-line/in-line monitoring of chemical and bio-chemical processes is generally difficult, time-consuming and costly. Therefore, it is preferable if calibration models can be used for an extended period, without the need to replace them. However, in many process applications, changes in the instrumental response (e.g. owing to a change of spectrometer) or variations in the measurement conditions (e.g. a change in temperature) can cause a multivariate calibration model to become invalid. In this contribution, a new method, systematic prediction error correction (SPEC), has been developed to maintain the predictive abilities of multivariate calibration models when e.g. the spectrometer or measurement conditions are altered. The performance of the method has been tested on two NIR data sets (one with changes in instrumental responses, the other with variations in experimental conditions) and the outcomes compared with those of some popular methods, i.e. global PLS, univariate slope and bias correction (SBC) and piecewise direct standardization (PDS). The results show that SPEC achieves satisfactory analyte predictions with significantly lower RMSEP values than global PLS and SBC for both data sets, even when only a few standardization samples are used. Furthermore, SPEC is simple to implement and requires less information than PDS, which offers advantages for applications with limited data.

Investigation and optimization of a passively operated compost-based system for remediation of acidic, highly iron- and sulfate-rich industrial waste water.

A passively operated multi-stage bioremediation system utilizing composted agricultural waste products and an artificial wetland system was found to be effective for purification of acidic, iron- and sulfate-rich waste water derived from titanium mineral processing. The main microbial players involved in the remediation system processes and the dynamics were investigated; mineral processing waste water-filled sludge dams possessed stable microbial communities that included Acidithiobacillus, Desulfurella, and acidophilic, anaerobic fermenters of the order Bacteroidales. These groups were enriched in a subsequent potato waste-based iron mobilization pre-treatment stage. Within downstream reduction treatment stages ("reduction cells"), compost/straw decomposition and associated sulfur/sulfate and iron reduction were carried out by a complex mix of aerobic and anaerobic bacteria. The efficaciousness of the system without replacement of the compost was found to steadily decline following 2 years of operation and corresponded with the reduction cell communities becoming simultaneously more diverse and homogenous. Microcosm-based experiments demonstrated that operational declines were due to unsustained supply of suitable labile carbon sources combined with spatial heterogeneity within the layered design of the reduction stage of the system resulting in inadequate redox conditions. Temperature was not found to be a critical performance factor in the range of 10-25 degrees C. Application of a combined emulsified oil/molasses amendment was found to be highly effective in promoting a microbial community capable of remediating waste water with high iron and sulfate levels. Acidophilic members of the order Bacteroidales were found to be critical in the investigated remediation system, providing organic donors for subsequent metal and sulfur transformations and could have a broader ecological significance than previously suspected.

In situ monitoring of the seed stage of a fermentation process using non-invasive NIR spectrometry.

Non-invasive NIR spectrometry has been used to monitor in situ the seed stage of a streptomyces fermentation process. The main spectral change occurred at 7263 cm(-1) in the 1st derivative spectrum, and from comparison with off-line NIR spectra acquired of components present in the fermentation broth, can be attributed to biomass. The biomass signal was constant for the first 20 h of the seed stage, after which it decreased before increasing again. The time at which the minimum occurred in the NIR profile was either the same or slightly earlier than that at which a maximum in the carbon dioxide evolution rate (CER) occurred. The changes observed for the biomass signal in the NIR spectra can be attributed to growth and then fragmentation of mycelia, which indicates a change in metabolic activity. Hence, it may be possible to use NIR spectrometry in situ to determine the optimum transfer time for the seed stage of a fermentation process. Spectra were also acquired of the final stage of the same fermentation process. The variation in the biomass signal in the NIR spectra was more complicated in the final stage owing to changes in stir rate, and biomass concentration and morphology.