Getting value out of your sensory testing - the difference from control test

Non-parametric difference tests such as triangle and duo-trio tests traditionally are used to establish differences or similarities between products. However they only supply the researcher with partial answers and often further testing is required to establish the nature, size and direction of differences. This paper looks at the advantages of the difference from control (DFC) test (also known as degree of difference test) and discusses appropriate applications of the test. The scope and principle of the test, panel composition and analysis of results are presented with the aid of suitable examples. Two of the major uses of the DFC test are in quality control and shelf-life testing. The role DFC takes in these areas and the use of other tests to complement the testing is discussed. Controls or standards are important in both these areas and the use of standard products, mental and written standards and blind controls are highlighted. The DFC test has applications in products where the duo-trio and triangle tests cannot be used because of the normal heterogeneity of the product. While the DFC test is a simple difference test it can be structured to give the researcher more valuable data and scope to make informed decisions about their product.

A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish bananas (Musa spp. AAA)

The dwarf somaclonal variant is a major problem affecting micropropagation of the banana cultivar Williams (Musa spp. AAA; subgroup Cavendish). This problem arises from genetic changes that occur during the tissue culture process. Early identification of this problem is difficult and propagators must wait until plants are ex vitro in order to visualise the dwarfism phenotype. In this study, we have improved a SCAR-based molecular diagnostic technique, developed by Damasco et al. [Acta Hortic. 461 (1997) 157], for the early identification of dwarf off-types. We have included a positive internal control in a multiplex PCR and adapted the technique for use with small amounts of fresh in vitro leaf material as PCR template. The control product is a 500 bp fragment from 18S rRNA and is amplified in all tissues irrespective of phenotype. The use of small in vitro leaf material removing the need for genomic DNA extraction.

Comparison of the indirect haemagglutination and gel diffusion test for serotyping Haemophilus parasuis

The aim of this study was to compare the use of indirect haemagglutination (IHA) and gel diffusion (GD) tests for serotyping Haemophilus parasuis by the Kielstein-Rapp-Gabrielson scheme. All 15 serovar reference strains, 72 Australian field isolates, nine Chinese field isolates, and seven isolates from seven experimentally infected pigs were evaluated with both tests. With the IHA test, 14 of the 15 reference strains were correctly serotyped – with serovar 10 failing to give a titre with serovar 10 antiserum. In the GD test, 13 reference strains were correctly serotyped – with antigen from serovars 7 and 8 failing to react with any antiserum. The IHA methodology serotyped a total of 45 of 81 field isolates while the GD methodology serotyped a total of 48 isolates. For 29 isolates, the GD and IHA methods gave discordant results. It was concluded that the IHA is a good additional test for the serotyping of H. parasuis by the KRG scheme if the GD methodology fails to provide a result or shows unusual cross-reactions.

A universal test to determine the integrity of RNA, and its suitability for reverse transcription, in animal tissue laboratory specimens

Degradation of RNA in diagnostic specimens can cause false-negative test results and potential misdiagnosis when tests rely on the detection of specific RNA sequence. Current molecular methods of checking RNA integrity tend to be host species or group specific, necessitating libraries of primers and reaction conditions. The objective here was to develop a universal (multi-species) quality assurance tool for determining the integrity of RNA in animal tissues submitted to a laboratory for analyses. Ribosomal RNA (16S rRNA) transcribed from the mitochondrial 16S rDNA was used as template material for reverse transcription to cDNA and was amplified using polymerase chain reaction (PCR). As mitochondrial DNA has a high level of conservation, the primers used were shown to reverse transcribe and amplify RNA from every animal species tested. Deliberate degradation of rRNA template through temperature abuse of samples resulted in no reverse transcription and amplification. Samples spiked with viruses showed that single-stranded viral RNA and rRNA in the same sample degraded at similar rates, hence reverse transcription and PCR amplification of 16S rRNA could be used as a test of sample integrity and suitability for analysis that required the sample's RNA, including viral RNA. This test will be an invaluable quality assurance tool for determination of RNA integrity from tissue samples, thus avoiding erroneous test results that might occur if degraded target RNA is used unknowingly as template material for reverse transcription and subsequent PCR amplification.

The vaccination-challenge trial: the gold standard test to evaluate the protective efficacy of infectious coryza vaccines

Infectious coryza is an upper respiratory tract disease of chickens with the major impact occurring in multi-age flocks. We investigated the relationship between the level of antibodies, as detected by a haemagglutination-inhibition (HI) assay, in infectious coryza-vaccinated chickens and the protection against challenge in those chickens. In one experiment, chickens given a single dose of either of two infectious coryza vaccines lacked a detectable HI response to vaccination but showed significant levels of protection 11 weeks after vaccination. In contrast, in chickens given two doses of an infectious coryza vaccine and challenged 3 weeks after the second vaccine dose, there was a strong serological response with 36/40 birds having a HI titre of 1/20 or greater. In this trial there was an apparent relationship between titre and subsequent protection, with none of the 32 chickens with a titre of 1/40 or 1/80 showing any clinical signs and only one of the same group yielding the challenge organism on culture. In contrast, three of the four vaccinated chickens with a HI titre less than 1/5 developed the typical clinical signs of coryza and yielded the challenge organism on culture. Overall, our results suggest that HI titres cannot be regarded as a definitive predictor of vaccine efficacy. We suggest that the vaccination-challenge trial is the gold standard for the evaluation of the immune response to infectious coryza vaccines.

Summative evaluation of climate application activities with pastoralists in western Queensland

Survey methods were engaged to measure the change in use and knowledge of climate information by pastoralists in western Queensland. The initial mail survey was undertaken in 2000-01 (n=43) and provided a useful benchmark of pastoralists climate knowledge. Two years of climate applications activities were completed and clients were re-surveyed in 2003 (n=49) to measure the change in knowledge and assess the effectiveness of the climate applications activities. Two methods were used to assess changes in client knowledge, viz., self-assessment and test questions. We found that the use of seasonal climate forecasts in decision making increased from 36% in 2001 (n=42) to 51% in 2003 (n=49) (P=0.07). The self-assessment technique was unsatisfactory as a measure of changing knowledge over short periods (1-3 years), but the test question technique was successful and indicated an improvement in climate knowledge among respondents. The increased levels of use of seasonal climate forecasts in management and improved knowledge was partly attributed to the climate applications activities of the project. Further, those who used seasonal forecasting (n=25) didn't understand key components of forecasts (e.g. probability, median) better than those who didn't use seasonal forecasts (n=24) (P>0.05). This identifies the potential for misunderstanding and misinterpretation of forecasts among users and highlights the need for providers of forecasts to understand the difficulties and prepare simply written descriptions of forecasts and disseminate these with the maps showing probabilities. The most preferred means of accessing climate information were internet, email, 'The Season Ahead' newsletter and newspaper. The least preferred were direct contact with extension officers and attending field days and group meetings. Eighty-six percent of respondents used the internet and 67% used ADSL broadband internet (April 2003). Despite these findings, extension officers play a key role in preparing and publishing the information on the web, in emails and newsletters. We also believe that direct contact with extension officers trained in climate applications is desirable in workshop-like events to improve knowledge of the difficult concepts underpinning climate forecasts, which may then stimulate further adoption.

Test of the enemy release hypothesis: the native magpie moth prefers a native fireweed (Senecio pinnatifolius) to its introduced congener (S. madagascariensis).

The enemy release hypothesis predicts that native herbivores will either prefer or cause more damage to native than introduced plant species. We tested this using preference and performance experiments in the laboratory and surveys of leaf damage caused by the magpie moth Nyctemera amica on a co-occuring native and introduced species of fireweed (Senecio) in eastern Australia. In the laboratory, ovipositing females and feeding larvae preferred the native S. pinnatifolius over the introduced S. madagascariensis. Larvae performed equally well on foliage of S. pinnatifolius and S. madagascariensis: pupal weights did not differ between insects reared on the two species, but growth rates were significantly faster on S. pinnatifolius. In the field, foliage damage was significantly greater on native S. pinnatifolius than introduced S. madagascariensis. These results support the enemy release hypothesis, and suggest that the failure of native consumers to switch to introduced species contributes to their invasive success. Both plant species experienced reduced, rather than increased, levels of herbivory when growing in mixed populations, as opposed to pure stands in the field; thus, there was no evidence that apparent competition occurred.

Simulating the evolution of glyphosate resistance in grains farming in northern Australia.

Background and Aims: The evolution of resistance to herbicides is a substantial problem in contemporary agriculture. Solutions to this problem generally consist of the use of practices to control the resistant population once it evolves, and/or to institute preventative measures before populations become resistant. Herbicide resistance evolves in populations over years or decades, so predicting the effectiveness of preventative strategies in particular relies on computational modelling approaches. While models of herbicide resistance already exist, none deals with the complex regional variability in the northern Australian sub-tropical grains farming region. For this reason, a new computer model was developed. Methods: The model consists of an age- and stage-structured population model of weeds, with an existing crop model used to simulate plant growth and competition, and extensions to the crop model added to simulate seed bank ecology and population genetics factors. Using awnless barnyard grass (Echinochloa colona) as a test case, the model was used to investigate the likely rate of evolution under conditions expected to produce high selection pressure. Key Results: Simulating continuous summer fallows with glyphosate used as the only means of weed control resulted in predicted resistant weed populations after approx. 15 years. Validation of the model against the paddock history for the first real-world glyphosate-resistant awnless barnyard grass population shows that the model predicted resistance evolution to within a few years of the real situation. Conclusions: This validation work shows that empirical validation of herbicide resistance models is problematic. However, the model simulates the complexities of sub-tropical grains farming in Australia well, and can be used to investigate, generate and improve glyphosate resistance prevention strategies.

Management Guidelines for New Warm-Season Grasses in Australia

After more than 30 years in which ‘Tifgreen’ and ‘Tifdwarf’ were the only greens-quality varieties available, the choice for golf courses and bowls clubs in northern Australia has been expanded to include six new Cynodon hybrids [Cynodon dactylon (L.) Pers x Cynodon transvaalensis Burtt-Davy]. Five of these – ‘Champion Dwarf’ (Texas), ‘MS-Supreme’ (Mississippi), FloraDwarf™ (Florida), ‘TifEagle’ (Georgia), MiniVerde™ (Arizona) - are from US breeding programs, while the sixth, ‘TL2’ (marketed as Novotek™) was selected in north Queensland. The finer, denser and lower growing habit of the “ultradwarf” cultivars allows very low mowing heights (e.g. 2.5 mm) to be imposed, resulting in denser and smoother putting and bowls surfaces. In addition to the Cynodon hybrids, four new greens quality seashore paspalum (Paspalum vaginatum O. Swartz) cultivars including ‘Sea Isle 2000’, Sea Isle Supreme™, Velvetene™ and Sea Dwarf™ (where tolerance of salty water is required) expands the range of choices for greens in difficult environments. The project was developed to determine (a) the appropriate choice of cultivar for different environments and budgets, and (b) best management practices for the new cultivars which differ from the Cynodon hybrid industry standards ‘Tifgreen’ and ‘Tifdwarf’. Management practices, particularly fertilising, mowing heights and frequency, and thatch control were investigated to determine optimum management inputs and provide high quality playing surfaces with the new grasses. To enable effective trialling of these new and old cultivars it was essential to have a number of regional sites participating in the study. Drought and financial hardship of many clubs presented an initial setback with numerous clubs wanting to be involved in the study but were unable to commit due to their financial position at the time. The study was fortunate to have seven regional sites from Queensland, New South Wales, Victoria and South Australia volunteer to be involved in the study which would add to the results being collected at the centralised test facility being constructed at DEEDI’s Redlands Research Station. The major research findings acquired from the eight trial sites included: • All of the new second generation “ultradwarf” couchgrasses tend to produce a large amount of thatch with MiniVerde™ being the greatest thatch producer, particularly compared to ‘Tifdwarf’ and ‘Tifgreen’. The maintenance of the new Cynodon hybrids will require a program of regular dethatching/grooming as well as regular light dustings of sand. Thatch prevention should begin 3 to 4 weeks after planting a new “ultradwarf” couchgrass green, with an emphasis on prevention rather than control. • The “ultradwarfs” produced faster green speeds than the current industry standards ‘Tifgreen’ and ‘Tifdwarf’. However, all Cynodon hybrids were considerably faster than the seashore paspalums (e.g. comparable to the speed diference of Bentgrass and couchgrass) under trial conditions. Green speed was fastest being cut at 3.5 mm and rolled (compared to 3.5 mm cut, no roll and 2.7 mm cut, no roll). • All trial sites reported the occurrence of disease in the Cynodon hybrids with the main incidence of disease occurring during the dormancy period (autumn and winter). The main disease issue reported was “patch diseases” which includes both Gaumannomyces and Rhizoctonia species. There was differences in the severity of the disease between cultivars, however, the severity of the disease was not consistent between cultivars and is largely attributed to an environment (location) effect. In terms of managing the occurrence of disease, the incidence of disease is less severe where there is a higher fertility rate (about 3 kgN/100m2/year) or a preventitatve fungicide program is adopted. • Cynodon hybrid and seashore paspalum cultivars maintained an acceptable to ideal surface being cut between 2.7 mm and 5.0 mm. “Ultradwarf” cultivars can tolerate mowing heights as low as 2.5 mm for short periods but places the plant under high levels of stress. Greens being maintained at a continually lower cutting height (e.g. 2.7 mm) of both species is achievable, but would need to be cut daily for best results. Seashore paspalums performed best when cut at a height of between 2.7 mm and 3.0 mm. If a lower cutting height is adopted, regular and repeated mowings are required to reduce scalping and produce a smooth surface. • At this point in time the optimum rate of nitrogen (N) for the Cynodon hybrids is 3 kg/100m2/year and while the seashore paspalums is 2 to 3 kg/100m2/year. • Dormancy occurred for all Cynodon and seashore paspalum culitvars from north in Brisbane (QLD) to south in Mornington Peninsula (VIC) and west to Novar Gardens (SA). Cynodon and Paspalum growth in both Victoria and South Australia was less favourable as a result of the cooler climates. • After combining the data collected from all eight sites, the results indicated that there can be variation (e.g. turfgrass quality, colour, disease resistance, performace) depending on the site and climatic conditions. Such evidence highlights the need to undertake genotype by environment (G x E) studies on new and old cultivars prior to conversion or establishment. • For a club looking to select either a Cynodon hybrid or seashore paspalum cultivar for use at their club they need to: - Review the research data. - Look at trial plots. - Inspect greens in play that have the new grasses. - Select 2 to 3 cultivars that are considered to be the better types. - Establish them in large (large enough to putt on) plots/nursery/practice putter. Ideally the area should be subjected to wear. - Maintain them exactly as they would be on the golf course/lawn bowls green. This is a critical aspect. Regular mowing, fertilising etc. is essential. - Assess them over at least 2 to 3 years. - Make a selection and establish it in a playing green so that it is subjected to typical wear.

Test and develop sustainable and profitable grazing strategies to manage for rainfall variability

The Wambiana grazing trial started in 1997 to test and develop sustainable and profitable grazing strategies to manage for rainfall variability. It is important that this trial continue as the results are still relatively short-term relative to rainfall cycles and significant treatment changes are still occurring. This new proposal will maintain baseline treatments but will modify others based on trial learning’s to date. It builds on treatment differences and evidence collected over the last 12 years to deliver evidence-based guidelines and principles for sustainable and productive management. The trial also links to other projects modelling water quality, climate change, methane emissions and soil C sequestration on grazing lands.

Development of commercial application of an avocado fruit robustness test

Fruit quality is one of the major factors limiting growth in avocado retail sales. Avocado growers are often unaware of their end-use fruit quality since quality problems only manifest upon fruit ripening and growers receive limited feedback from the supply chain. If growers were aware of their expected fruit quality they would be equipped to make better marketing decisions and if necessary to take remedial actions to improve their fruit quality. Avotest is being developed as a quick and easy method of determining expected end-use fruit quality before the start of the commercial fruit harvest. The test aims at distinguishing between blocks with robust fruit and those with less robust fruit. The test could also be used to predict the resulting fruit quality after the implementation of new farming practices.

Corymbia leaf oils, latitude, hybrids and herbivory: a test using common-garden field trials

Foliar oils, particularly monoterpenes, can influence the susceptibility of plants to herbivory. In plants, including eucalypts, monoterpenes are often associated with plant defence. A recent analysis revealed an increase in foliar oil content with increasing latitudinal endemism, and we tested this pattern using three eucalypt taxa comprising a latitudinal replacement cline. We also examined the relative concentrations of two monoterpenes (alpha-pinene and 1,8-cineole), for which meta-analyses also showed latitudinal variation, using hybrids of these three taxa with Corymbia torelliana. These, and pure C. torelliana, were then assessed in common-garden field plots for the abundance and distribution of herbivory by four distinct herbivore taxa. Differing feeding strategies among these herbivores allowed us to test hypotheses regarding heritability of susceptibility and relationships to alpha-pinene and 1,8-cineole. We found no support for an increase in foliar oil content with increasing latitude, nor did our analysis support predictions for consistent variation in alpha-pinene and 1,8-cineole contents with latitude. However, herbivore species showed differential responses to different taxa and monoterpene contents. For example, eriophyid mites, the most monophagous of our censused herbivores, avoided the pure species, but fed on hybrid taxa, supporting hypotheses on hybrid susceptibility. The most polyphagous herbivore (leaf blister sawfly Phylacteophaga froggatti) showed no evidence of response to plant secondary metabolites, while the distribution and abundance patterns of Paropsis atomaria showed some relationship to monoterpene yields.

Preference and performance of Diachasmimorpha kraussii (Fullaway) (Hymenoptera: Braconidae) on five commercial fruit species

Diachasmimorpha kraussii is a polyphagous endoparasitoid of dacine fruit flies. The fruit fly hosts of D. krausii, in turn, attack a wide range of fruits and vegetables. The role that fruits play in host selection behaviour of D. kraussii has not been previously investigated. This study examines fruit preference of D. kraussii through a laboratory choice-test trial and field fruit sampling. In the laboratory trial, oviposition preference and offspring performance measures (sex ratio, developmental time, body length, hind tibial length) of D. kraussii were investigated with respect to five fruit species [Psidium guajava L. (guava), Prunis persica L. (peach), Malus domestica Borkh. (apple), Pyrus communis L. (pear) and Citrus sinensis L. (orange)], and two fruit fly species (Bactrocera jarvisi and B. tryoni). Diachasmimorpha kraussii responded to infested fruit of all fruit types in both choice and no-choice tests, but showed stronger preference for guava and peach in the choice tests irrespective of the species of fly larvae within the fruit. The wasp did not respond to uninfested fruit. The offspring performance measures differed in a non-consistent fashion between the fruit types, but generally wasp offspring performed better in guava, peach and orange. The offspring sex ratio, except for one fruit/fly combination (B. jarvisi in apple), was always female biased. The combined results suggest that of the five fruits tested, guava and peach are the best fruit substrates for D. krausii. Field sampling indicated a non-random use of available, fruit fly infested fruit by D. kraussii. Fruit fly maggots within two fruit species, Plachonia careya and Terminalia catappa, had disproportionately higher levels of D. krausii parasitism than would be expected based on the proportion of different infested fruit species sampled, or levels of fruit fly infestation within those fruit.

Preliminary assessment of a rapid leaf nitrogen test in mango

Nitrogen (N) is an essential nutrient in mango, influencing both productivity and fruit quality. In Australia, tree N is traditionally assessed once a year in the dormant pre-flowering stage by laboratory analysis of leaf N. This single assessment is insufficient to determine tree N status at all stages of the annual phenological cycle. Development of a field-based rapid N test would allow more frequent monitoring of tree N status and improved fertiliser management. This experiment examined the accuracy and useability of several devices used in other horticultural crops to rapidly assess mango leaf N in the field; the Konica Minolta 'SPAD-502 chlorophyll meter', Horiba 'Cardy Meter' and the Merck 'RQflex 10'. Regression and correlation analyses were used to determine the relationship between total leaf N and the measurements from the rapid test devices. The relationship between the chlorophyll index measured by the SPAD-502 meter and leaf N is highly significant at late fruit set (R 2=0.72, n=40) and post-harvest (R2=0.81, n=40) stages in the mango cultivar 'Kensington Pride' and significant (R2=0.51, n=40) at the flowering stage, indicating the device can be used to rapidly assess mango leaf N in the field. Correlation analysis indicated the relationship between petiole sap measured with the Cardy or Merck devices and leaf N is non-significant. © 2013 ISHS.

Sources of vase life variation in cut roses: A review

n determining vase life (VL), it is often not considered that the measured VL in a particular experiment may greatly depend on both the preharvest and evaluation environmental conditions. This makes the comparison between studies difficult and may lead to erroneous interpretation of results. In this review, we critically discuss the effect of the growth environment on the VL of cut roses. This effect is mainly related to changes in stomatal responsiveness, regulating water loss, whereas cut flower carbohydrate status appears less critical. When comparing cultivars, postharvest water loss and VL often show no correlation, indicating that components such as variation in the tissue resistance to cavitate and/or collapse at low water potential play an important role in the incidence of water stress symptoms. The effect of the growth environment on these components remains unknown. Botrytis cinerea sporulation and infection, as well as cut rose susceptibility to the pathogen are also affected by the growth environment, with the latter being largely unexplored. A huge variability in the choices made with respect to the experimental setup (harvest/conditioning methods, test room conditions and VL terminating symptoms) is reported. We highlight that these decisions, though frequently overlooked, influence the outcome of the study. Specifications for each of these factors are proposed as necessary to achieve a common VL protocol. Documentation of both preharvest conditions and a number of postharvest factors, including the test room conditions, is recommended not only for assisting comparisons between studies, but also to identify factors with major effects on VL.