Pasteurella multocida Expresses Two Lipopolysaccharide Glycoforms Simultaneously, but Only a Single Form Is Required for Virulence: Identification of Two Acceptor-Specific Heptosyl I Transferases

Lipopolysaccharide (LPS) is a critical virulence determinant in Pasteurella multocida and a major antigen responsible for host protective immunity. In other mucosal pathogens, variation in LPS or lipooligosaccharide structure typically occurs in the outer core oligosaccharide regions due to phase variation. P. multocida elaborates a conserved oligosaccharide extension attached to two different, simultaneously expressed inner core structures, one containing a single phosphorylated 3-deoxy-D-manno-octulosonic acid (Kdo) residue and the other containing two Kdo residues. We demonstrate that two heptosyltransferases, HptA and HptB, add the first heptose molecule to the Kdo1 residue and that each exclusively recognizes different acceptor molecules. HptA is specific for the glycoform containing a single, phosphorylated Kdo residue (glycoform A), while HptB is specific for the glycoform containing two Kdo residues (glycoform B). In addition, KdkA was identified as a Kdo kinase, required for phosphorylation of the first Kdo molecule. Importantly, virulence data obtained from infected chickens showed that while wild-type P. multocida expresses both LPS glycoforms in vivo, bacterial mutants that produced only glycoform B were fully virulent, demonstrating for the first time that expression of a single LPS form is sufficient for P. multocida survival in vivo. We conclude that the ability of P. multocida to elaborate alternative inner core LPS structures is due to the simultaneous expression of two different heptosyltransferases that add the first heptose residue to the nascent LPS molecule and to the expression of both a bifunctional Kdo transferase and a Kdo kinase, which results in the initial assembly of two inner core structures.

Spot Form of Net Blotch Resistance in a Diverse Set of Barley Lines in Australia and Canada.

The responses of 95 barley lines and cultivars to spot form of net blotch (SFNB) caused by Pyrenophora teres f. maculata were analyzed as seedlings and adults in Australia and Canada. Cluster analyses revealed complex reaction responses. Only 2 lines (Esperance Orge 289 and TR3189) were resistant to all isolates at the seedling stage, whereas 15 lines and cultivars (81-82/033, Arimont, BYDV-018, CBSS97M00855T-B2-M1-Y1-M2-Y-1M-0Y, C19776, Keel, Sloop, Torrens, TR326, VB0111, Yarra, VB0229, WI-2477, WI2553, and Wisconsin Pedigree) were resistant toward the two Canadian isolates and mixture of Australian isolates at the adult stages. In Australian field experiments, the effectiveness of SFNB resistance in three barley cultivars (Barque. Cowabbie, and Schooner) and one breeding line (VB9104) with a different source of resistance was tested. Barque, which possessed a resistance gene that provided complete resistance to SFNB, was the most effective and showed no effect on grain yield or quality in the presence of inoculum. Generally, cultivars with seedling or adult resistance had less disease and better grain quality than the susceptible control. Dash, but they were not as effective as Barque. A preliminary differential set of 19 barley lines and cultivars for P teres I. maculata is proposed.

Influence of environmental and instrumental variables on the non-invasive prediction of Brix in pineapple using near infrared spectroscopy

The Brix content of pineapple fruit can be non-invasively predicted from the second derivative of near infrared reflectance spectra. Correlations obtained using a NIRSystems 6500 spectrophotometer through multiple linear regression and modified partial least squares analyses using a post-dispersive configuration were comparable with that from a pre-dispersive configuration in terms of accuracy (e.g. coefficient of determination, R2, 0.73; standard error of cross validation, SECV, 1.01°Brix). The effective depth of sample assessed was slightly greater using the post-dispersive technique (about 20 mm for pineapple fruit), as expected in relation to the higher incident light intensity, relative to the pre-dispersive configuration. The effect of such environmental variables as temperature, humidity and external light, and instrumental variables such as the number of scans averaged to form a spectrum, were considered with respect to the accuracy and precision of the measurement of absorbance at 876 nm, as a key term in the calibration for Brix, and predicted Brix. The application of post-dispersive near infrared technology to in-line assessment of intact fruit in a packing shed environment is discussed.

Discrepancy Between Antennal and Behavioral Responses for Enantiomers of a-Pinene: Electrophysiology and Behavior of Helicoverpa Armigera (Lepidoptera)

The ability of adult cotton bollworm, Helicoverpa armigera (Hubner), to distinguish and respond to enantiomers of α-pinene was investigated with electrophysiological and behavioral methods. Electroantennogram recordings using mixtures of the enantiomers at saturating dose levels, and single unit electrophysiology, indicated that the two forms were detected by the same receptor neurons. The relative size of the electroantennogram response was higher for the (−) compared to the (+) form, indicating greater affinity for the (−) form at the level of the dendrites. Behavioral assays investigated the ability of moths to discriminate between, and respond to the (+) and (−) forms of α pinene. Moths with no odor conditioning showed an innate preference for (+)-α-pinene. This preference displayed by naıve moths was not significantly different from the preferences of moths conditioned on (+)-α-pinene. However, we found a significant difference in preference between moths conditioned on the (−) enantiomer compared to naıve moths and moths conditioned on (+)-α-pinene, showing that learning plays an important role in the behavioral response. Moths are less able to distinguish between enantiomers of α-pinene than different odors (e.g., phenylacetaldehyde versus (−)-α-pinene) in learning experiments. The relevance of receptor discrimination of enantiomers and learning ability of the moths in host plant choice is discussed.

Chemical Characterization of Urinary Pheromones in Brown Antechinus, Antechinus stuartii

In the small dasyurid marsupial, Antechinus stuartii, males exhibit scent-marking in the form of cloacal marking of nesting areas during the breeding season. Females of this species show no such behavior. To characterize the potential male pheromonal scent signal, urine-derived volatiles from sexually active males were analyzed by GC-MS and compared to that of females and a castrated male. More than 10 urinary compounds were identified. A series of homologous methylketones was observed in both males and females, whereas aldehydes were present only in female urine. Urine from the castrate was virtually compound-free except for minute concentrations of a compound tentatively identified as 2,4-dithiapentane. This compound was also found in one of the sexually active males. The GC profiles of the sexually active males contained high concentrations of two pyrazine derivatives and four methylketones that were not detected in the profiles of either females or the castrate. These compounds may influence social communication in the brown antechinus, Antechinus stuartii.

The process for implementation of a Quality Management System within a multi-functional cereal laboratory

The intent of this study was to design, document and implement a Quality Management System (QMS) into a laboratory that incorporated both research and development (R&D) and routine analytical activities. In addition, it was necessary for the QMS to be easily and efficiently maintained to: (a) provide documented evidence that would validate the system's compliance with a certifiable standard, (b) fit the purpose of the laboratory, (c) accommodate prevailing government policies and standards, and (d) promote positive outcomes for the laboratory through documentation and verification of the procedures and methodologies implemented. Initially, a matrix was developed that documented the standards' requirements and the necessary steps to be made to meet those requirements. The matrix provided a check mechanism on the progression of the system's development. In addition, it was later utilised in the Quality Manual as a reference tool for the location of full procedures documented elsewhere in the system. The necessary documentation to build and monitor the system consisted of a series of manuals along with forms that provided auditable evidence of the workings of the QMS. Quality Management (QM), in one form or another, has been in existence since the early 1900's. However, the question still remains: is it a good thing or just a bugbear? Many of the older style systems failed because they were designed by non-users, fiercely regulatory, restrictive and generally deemed to be an imposition. It is now considered important to foster a sense of ownership of the system by the people who use the system. The system's design must be tailored to best fit the purpose of the operations of the facility if maximum benefits to the organisation are to be gained.

Glucosinolate composition and anti-cancer potential of daikon and radish sprouts

Daikon and radish sprouts contain high levels of glucoraphenin, a glucosinolate which hydrolyses to form sulphoraphene. Sulphoraphene, like sulphoraphane from broccoli, is a potent inducer of phase 2 detoxification enzymes and consequently has potential anti-cancer action. Unlike broccoli however, daikon and radish do not possess epithiospecifier protein, a protein that inhibits conversion of glucosinolates to isothiocyanates, and consequently they may represent more suitable sources of phyto-chemicals with anti-cancer potential. Concentrations of glucoraphenin were highest in the seed, declining exponentially with sprout development. The rate of decline was observed to vary considerably between varieties of daikon and radish, with some varieties maintaining significantly high levels of glucoraphenin. Varieties maintaining a high level of glucoraphenin included 'Cherry Belle' and 'French Breakfast'.

Floating row covers and pyriproxyfen help control silverleaf whitefly Bemisia tabaci (Gennadius) Biotype B (Homoptera: Aleyrodidae) in zucchini

In zucchini, the use of row covers until flowering and the insect growth regulator (IGR) pyriproxyfen are effective methods of reducing the number of insects, especially silverleaf whitefly (Bemisia tabaci (Gennadius) Biotype B), on plants. We compared floating row covers (FRCs) up until flowering with silverleaf whitefly (SLW) introduced (FRC + SLW), or not introduced (FRC-only), or with introduction of SLW in open plots (SLW-only), or with introduction of SLW in open plots with IGR (SLW + IGR). FRC increased temperature and humidity compared with the uncovered treatments. Average fruit weight was less (P < 0.01) for the FRC + SLW treatment compared with the other treatments and the percentage of marketable fruit was less for the FRC + SLW than for the other three treatments. This result indicates that the use of either row covers or IGR controls whiteflies, reduces fruit damage and increases the size, weight, and quality of fruit, and may also control other sap-sucking insects. However, if SLW are already present on plants, the use of FRC may reduce predation and favour build up of SLW. Thus, FRC and IGR, if used judiciously, may provide an effective alternative to broad-spectrum pesticides in small-scale cucurbit production.

Identification of small juvenile scombrids from northwest tropical Australia using mitochondrial DNA cytochrome b sequences

Small juveniles of the nine species of scombrids in Australian waters are morphologically similar to one another and, consequently, difficult to identify to species level. We show that the sequence of the mitochondrial DNA cytochrome b gene region is a powerful tool for identification of these young fish. Using this method, we identified 50 juvenile scombrids collected from Exmouth Bay, Western Australia. Six species of scombrids were apparent in this sample of fish: narrow-barred Spanish mackerel (Scomberomorus commerson), Indian mackerel (Rastrelliger kanagurta), frigate tuna (Auxis thazard), bullet tuna (Auxis rochei), leaping bonito (Cybiosarda elegans), and kawakawa (Euthynnus affinis). The presence of Indian mackerel, frigate tuna, leaping bonito, and kawakawa is the first indication that coastal waters may be an important spawning habitat for these species, although offshore spawning may also occur. The occurrence of small juvenile S. commerson was predicted from the known spawning patterns of that species, but other mackerel species (Scomberomorus munroi, Scomberomorus queenslandicus, Scomberomorus semifasiciatus) likely to be spawning during the sampling period were not detected among the 50 small juveniles analyzed here.

Sampling and management of Bemisia tabaci (Genn.) biotype B in Australian cotton

Data on seasonal population abundance of Bemisia tabaci biotype B (silverleaf whitefly (SLW)) in Australian cotton fields collected over four consecutive growing seasons (2002/2003-2005/2006) were used to develop and validate a multiple-threshold-based management and sampling plan. Non-linear growth trajectories estimated from the field sampling data were used as benchmarks to classify adult SLW field populations into six density-based management zones with associated control recommendations in the context of peak flowering and open boll crop growth stages. Control options based on application of insect growth regulators (IGRs) are recommended for high-density populations (>2 adults/leaf) whereas conventional (non-IGR) products are recommended for the control of low to moderate population densities. A computerised re-sampling program was used to develop and test a binomial sampling plan. Binomial models with thresholds of T=1, 2 and 3 adults/leaf were tested using the field abundance data. A binomial plan based on a tally threshold of T=2 adults/leaf and a minimum sample of 20 leaves at nodes 3, 4 or 5 below the terminal is recommended as the most parsimonious and practical sampling protocol for Australian cotton fields. A decision support guide with management zone boundaries expressed as binomial counts and control options appropriate for various SLW density situations is presented. Appropriate use of chemical insecticides and tactics for successful field control of whiteflies are discussed.

Light environment and tree development of young Acacia melanoxylon in mixed-species regrowth forest, Tasmania, Australia

Blackwood (Acacia melanoxylon R. Br.) is a valuable leguminous cabinetwood species which is commonly found as a canopy or subcanopy tree in a broad range of mixed-species moist forests on tablelands and coastal escarpments in eastern Australia. This paper reports on the competitive light environment of a commercially valuable multi-species regrowth forest in NW Tasmania, in order to define some of the functional interactions and competitive dynamics of these stands. Comparative observations were made of the internal forest light environment in response to small-gap silvicultural treatments, in a young regenerative mix of three codominant tree species. Light measurements were made during periods of maximum external irradiance of the regrowth Eucalyptus obliqua/A. melanoxylon forest canopy at age 10.5 years. This was at a time of vigourous stand development, 4.5 years following the application of three experimental silvicultural treatments whose effects were observed in comparison with an untreated canopy sample designed as a control. Minimal irradiance was observed within and beneath the dense subcanopy of the native nurse species (Pomaderris apetala) which closely surrounds young blackwood regeneration. Unlike current plantation nurse systems, the dense foliage of the native broadleaved Pomaderris all but eliminated direct side-light and low-angle illumination of the young blackwood, from the beginning of tree establishment. The results demonstrated that retention of these densely stocked native codominants effectively suppressed both size and frequency of blackwood branches on the lower bole, through effective and persistent interception of sunlight. Vigorous young blackwood crowns later overtopped the codominant nurse species, achieving a predictable height of branch-free bole. This competitive outcome offers a valuable tool for management of blackwood crown dynamics, stem form and branch habit through manipulation of light environment in young native regrowth systems. Results demonstrate that effective self-pruning in the lower bole of blackwood is achieved through a marked reduction in direct and diffuse sunlight incident on the lower crown, notably to less than 10-15% of full sunlight intensity during conditions of maximum insolation. The results also contain insights for the improved design of mixed-species plantation nurse systems using these or functionally similar species' combinations. Based on evidence presented here for native regrowth forest, plantation nurse systems for blackwood will need to achieve 85-90% interception of external side-light during early years of tree development if self-pruning is to emulate the results achieved in the native nurse system.

Perennial grassland dynamics on fertile plains: Is coexistence mediated by disturbance?

The response of grasslands to disturbance varies with the nature of the disturbance and the productivity of the landscape. In highly productive grasslands, competitive exclusion often results in decreased species richness and grazing may allow more species to coexist. Once widespread, grasslands dominated by Dichanthium sericeum (Queensland bluegrass) and Astrebla spp. (Mitchell grass) occur on fertile plains but have been reduced in extent by cultivation. We tested the effects of exclusion of livestock grazing on these grasslands by comparing the floristic composition of sites in a nature reserve with an adjacent stock reserve. In addition, sites that had been cultivated within the nature reserve were compared with those where grazing but no cultivation had occurred. To partition the effects of temporal variation from spatial variation we sampled sites in three different years (1998, 2002 and 2004). Some 194 taxa were recorded at the nature reserve and surrounding stock routes. Sampling time, the occurrence of past cultivation and livestock grazing all influenced species composition. Species richness varied greatly between sampling periods relating to highly variable rainfall and water availability on heavy clay soils. Native species richness was significantly lower at previously cultivated sites (13-22 years after cultivation), but was not significantly influenced by grazing exclusion. After 8 years it appears that reintroducing disturbance in the form of livestock grazing is not necessary to maintain plant species richness in the reserve. The highly variable climate (e.g. droughts) probably plays an important role in the coexistence of species by negating competitive exclusion and allowing interstitial species to persist.

The accuracy of varietal selection using factor analytic models for multi-environment plant breeding trials

Modeling of cultivar x trial effects for multienvironment trials (METs) within a mixed model framework is now common practice in many plant breeding programs. The factor analytic (FA) model is a parsimonious form used to approximate the fully unstructured form of the genetic variance-covariance matrix in the model for MET data. In this study, we demonstrate that the FA model is generally the model of best fit across a range of data sets taken from early generation trials in a breeding program. In addition, we demonstrate the superiority of the FA model in achieving the most common aim of METs, namely the selection of superior genotypes. Selection is achieved using best linear unbiased predictions (BLUPs) of cultivar effects at each environment, considered either individually or as a weighted average across environments. In practice, empirical BLUPs (E-BLUPs) of cultivar effects must be used instead of BLUPs since variance parameters in the model must be estimated rather than assumed known. While the optimal properties of minimum mean squared error of prediction (MSEP) and maximum correlation between true and predicted effects possessed by BLUPs do not hold for E-BLUPs, a simulation study shows that E-BLUPs perform well in terms of MSEP.

Stocking density and artificial habitat influence stock structure and yield from intensive nursery systems for mud crabs Scylla serrata (Forsskål 1775)

Intensive nursery systems are designed to culture mud crab postlarvae through a critical phase in preparation for stocking into growout systems. This study investigated the influence of stocking density and provision of artificial habitat on the yield of a cage culture system. For each of three batches of postlarvae, survival, growth and claw loss were assessed after each of three nursery phases ending at crab instars C1/C2, C4/C5 and C7/C8. Survival through the first phase was highly variable among batches with a maximum survival of 80% from megalops to a mean crab instar of 1.5. Stocking density between 625 and 2300 m-2 did not influence survival or growth in this first phase. Stocking densities tested in phases 2 and 3 were 62.5, 125 and 250 m -2. At the end of phases 2 and 3, there were five instar stages present, representing a more than 20-fold size disparity within the populations. Survival became increasingly density-sensitive following the first phase, with higher densities resulting in significantly lower survival (phase 2: 63% vs. 79%; phase 3: 57% vs. 64%). The addition of artificial habitat in the form of pleated netting significantly improved survival at all densities. The mean instar attained by the end of phase 2 was significantly larger at a lower stocking density and without artificial habitat. No significant effect of density or habitat on harvest size was detected in phase 3. The highest incidence of claw loss was 36% but was reduced by lowering stocking densities and addition of habitat. For intensive commercial production, yield can be significantly increased by addition of a simple net structure but rapidly decreases the longer crablets remain in the nursery.

Mapping of adult plant resistance to net form of net blotch in three Australian barley populations

Net form of net blotch (NFNB), caused by Pyrenophora teres Drechs. f. teres Smedeg., is a serious disease problem for the barley industry in Australia and other parts of the world. Three doubled haploid barley populations, Alexis/Sloop, WI2875-1/Alexis, and Arapiles/Franklin, were used to identify genes conferring adult plant resistance to NFNB in field trials. Quantitative trait loci (QTLs) identified were specific for adult plant resistance because seedlings of the parental lines were susceptible to the NFNB isolates used in this study. QTLs were identified on chromosomes 2H, 3H, 4H, and 7H in both the Alexis/Sloop and WI2875-1/Alexis populations and on chromosomes 1H, 2H, and 7H in the Arapiles/Franklin population. Using QTLNetwork, epistatic interactions were identified between loci on chromosomes 3H and 6H in the Alexis/Sloop population, between 2H and 4H in the WI2875-1/Alexis population, and between 5H and 7H in the Arapiles/Franklin population. Comparisons with earlier studies of NFNB resistance indicate the pathotype-dependent nature of many resistance QTLs and the importance of establishing an international system of pathotype nomenclature and differential testing.