Cyanidin 3-glucoside improves diet-induced metabolic syndrome in rats

Increased consumption of dark-coloured fruits and vegetables may mitigate metabolic syndrome. This study has determined the changes in metabolic parameters, and in cardiovascular and liver structure and function, following chronic administration of either cyanidin 3-glucoside (CG) or Queen Garnet plum juice (QG) containing cyanidin glycosides to rats fed either a corn starch (C) or a high-carbohydrate, high-fat (H) diet. Eight to nine-week-old male Wistar rats were randomly divided into six groups for 16-week feeding with C, C with CG or QG, H or H with CG or QG. C or H were supplemented with CG or QG at a dose of ∼8 mg/kg/day cyanidin glycosides from week 8 to 16. H rats developed signs of metabolic syndrome including visceral adiposity, impaired glucose tolerance, hypertension, cardiovascular remodelling, increased collagen depots in left ventricle, non-alcoholic fatty liver disease, increased plasma liver enzymes and increased inflammatory cell infiltration in the heart and liver. Both CG and QG reversed these cardiovascular, liver and metabolic signs. However, no intact anthocyanins or common methylated/conjugated metabolites could be detected in the plasma samples and plasma hippuric acid concentrations were unchanged. Our results suggest CG is the most likely mediator of the responses to QG but that further investigation of the pharmacokinetics of oral CG in rats is required.

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.

Phosphorus nutrition and tolerance of cotton to water stress: II. Water relations, free and bound water and leaf expansion rate

In previous experiments, increased leaf-Phosphorus (P) content with increasing P supply enhanced the individual leaf expansion and water content of fresh cotton leaves in a severely drying soil. In this paper, we report on the bulk water content of leaves and its components, free and bound water, along with other measures of plant water status, in expanding cotton leaves of various ages in a drying soil with different P concentrations. The bound water in living tissue is more likely to play a major role in tolerance to abiotic stresses by maintaining the structural integrity and/or cell wall extensibility of the leaves, whilst an increased amount of free water might be able to enhance solute accumulation, leading to better osmotic adjustment and tolerance to water stress, and maintenance of the volumes of sub-cellular compartments for expansive leaf growth. There were strong correlations between leaf-P%, leaf water (total, free and bound water) and leaf expansion rate (LER) under water stress conditions in a severely drying soil. Increased soil-P enhanced the uptake of P from a drying soil, leading to increased supply of osmotically active inorganic solutes to the cells in growing leaves. This appears to have led to the accumulation of free water and more bound water, ultimately leading to increased leaf expansion rates as compared to plants in low P soil under similar water stress conditions. The greater amount of bound and free water in the high-P plants was not necessarily associated with changes in cell turgor, and appears to have maintained the cell-wall properties and extensibility under water stressed conditions in soils that are nutritionally P-deficient.

Phosphorus nutrition and tolerance of cotton to water stress I. Seed cotton yield and leaf morphology

Seed cotton yield and morphological changes in leaf growth were examined under drying soil with different phosphorus (P) concentrations in a tropical climate. Frequent soil drying is likely to induce a decrease in nutrients particularly P due to reduced diffusion and poor uptake, in addition to restrictions in available water, with strong interactive effects on plant growth and functioning. Increased soil P in field and in-ground soil core studies increased the seed cotton yield and related morphological growth parameters in a drying soil, with hot (daily maximum temperature >33°C) and dry conditions (relative humidity, 25% to 35%), particularly during peak boll formation and filling stage. The soil water content in the effective rooting zone (top 0.4 m) decreased to -1.5 MPa by day 5 of the soil drying cycle. However, the increased seed cotton yield for the high-P plants was closely related to increasing leaf area with increased P supply. Plant height, leaf fresh mass and leaf area per plant were positively related to the leaf P%, which increased with increasing P supply. Low P plants were lower in plant height, leaf area, and leaf tissue water in the drying soil. Individual leaf area and the water content of the fresh leaf (ratio of dry mass to fresh mass) were significantly dependent on leaf P%.

Measurement and Interpretation of Salinity Tolerance in Four Perennial Grasses

Whilst the topic of soil salinity has received a substantive research effort over the years, the accurate measurement and interpretation of salinity tolerance data remain problematic. The tolerance of four perennial grass species (non-halophytes) to sodium chloride (NaCl) dominated salinity was determined in a free-flowing sand culture system. Although the salinity tolerance of non-halophytes is often represented by the threshold salinity model (bent-stick model), none of the species in the current study displayed any observable salinity threshold. Further, the observed yield decrease was not linear as suggested by the model. On re-examination of earlier datasets, we conclude that the threshold salinity model does not adequately describe the physiological processes limiting growth of non-halophytes in saline soils. Therefore, the use of the threshold salinity model is not recommended for non-halophytes, but rather, a model which more accurately reflects the physiological response observed in these saline soils, such as an exponential regression curve.

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.

Thermal tolerance and potential distribution of Carvalhotingis visenda (Hemiptera: Tingidae), a biological control agent for cat's claw creeper, Macfadyena unguis-cati (Bignoniaceae)

The specialist tingid, Carvalhotingis visenda, is a biological control agent for cat's claw creeper, Macfadyena unguis-cati (Bignoniaceae). Cat's claw creeper is an invasive liana with a wide climatic tolerance, and for biological control to be effective the tingid must survive and develop over a range of temperatures. We evaluated the effect of constant temperatures (0-45°C) on the survival and development of C. visenda. Adults showed tolerance for wider temperature ranges (0-45°C), but oviposition, egg hatching and nymphal development were all affected by both high (>30°C) and low (<20°C) temperatures. Temperatures between 20°C and 30°C are the most favourable for adult survival, oviposition, egg hatching and nymphal development. The ability of adults and nymphs to survive for a few days at high (40°C and 45°C) and low (0°C and 5°C) temperatures suggest that extreme temperature events, which usually occur for short durations (hours) in cat's claw creeper infested regions in Queensland and New South Wales states are not likely to affect the tingid population. The potential number of generations (egg to adult) the tingid can complete in a year in Australia ranged from three to eight, with more generations in Queensland than in New South Wales.

Salinity tolerance of twelve hybrid Bermudagrass (Cynodon Dactylon (L.) Pers. x C. Transvaalensis Burtt Davy) genotypes

Salinity is an increasingly important issue in both rural and urban areas throughout much of Australia. The use of recycled/reclaimed water and other sources of poorer quality water to irrigate turf is also increasing. Hybrid Bermudagrass (Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt Davey), together with the parent species C. dactylon, are amongst the most widely used warm-season turf grass groups. Twelve hybrid Bermudagrass genotypes and one accession each of Bermudagrass (C. dactylon), African Bermudagrass (C. transvaalensis) and seashore paspalum (Paspalum vaginatum Sw.) were grown in a glasshouse experiment with six different salinity treatments applied hydroponically through the irrigation water (ECW = <0.1, 6, 12, 18, 24 or 30 dSm-1) in a flood-and-drain system. Each pot was clipped progressively at 2-weekly intervals over the 12-week experimental period to determine dry matter production; leaf firing was rated visually on 3 occasions during the last 6 weeks of salinity treatment. At the end of the experiment, dry weights of roots and crowns below clipping height were also determined. Clipping yields declined sharply after about the first 6 weeks of salinity treatment, but then remained stable at substantially lower levels of dry matter production from weeks 8 to 12. Growth data over this final 4-week experimental period is therefore a more accurate guide to the relative salinity tolerance of the 15 entries than data from the preceding 8 weeks. Based on these data, the 12 hybrid Bermudagrass genotypes showed moderate salinity tolerance, with FloraDwarfM, 'Champion Dwarf', NovotekM and 'TifEagle' ranking as the most salt tolerant and 'Patriot', 'Santa Ana', 'Tifgreen' and TifSport M the least tolerant within the hybrid group. Nevertheless, Santa Ana, for example, maintained relatively strong root growth as salinity increased, and so may show better salt tolerance in practice than predicted from the growth data alone. The 12 hybrid Bermudagrasses and the single African Bermudagrass genotype were all ranked above FloraTeXM Bermudagrass in terms of salt tolerance. However, seashore paspalum, which is widely acknowledged as a halophytic species showing high salt tolerance, ranked well above all 14 Cynodon genotypes in terms of salinity tolerance.

Factors contributing to wear tolerance of Bermudagrass (Cynodon dactylon (L.) Pers., C. dactylon x C. transvaalensis Burtt-Davey) on a sand-based profile under simulated sports field conditions

Wear resistance and recovery of 8 Bermudagrass (Cynodon dactylon (L.) Pers.) and hybrid Bermudagrass (C. Dactylon x C. transvaalensis Burtt-Davey) cultivars grown on a sandbased soil profile near Brisbane, Australia, were assessed in 4 wear trials conducted over a two year period. Wear was applied on a 7-day or a 14-day schedule by a modified Brinkman Traffic Simulator for 6-14 weeks at a time, either during winter-early spring or during summer-early autumn. The more frequent wear under the 7-day treatment was more damaging to the turf than the 14-day wear treatment, particularly during winter when its capacity for recovery from wear was severely restricted. There were substantial differences in wear tolerance among the 8 cultivars investigated, and the wear tolerance rankings of some cultivars changed between years. Wear tolerance was associated with high shoot density, a dense stolon mat strongly rooted to the ground surface, high cell wall strength as indicated by high total cell wall content, and high levels of lignin and neutral detergent fiber. Wear tolerance was also affected by turf age, planting sod quality, and wet weather. Resistance to wear and recovery from wear are both important components of wear tolerance, but the relative importance of their contributions to overall wear tolerance varies seasonally with turf growth rate.

Physiological Basis for Genotypic Variation in Tolerance to and Recovery from Pre-flowering Drought in Peanut.

Drought during the pre-flowering stage can increase yield of peanut. There is limited information on genotypic variation for tolerance to and recovery from pre-flowering drought (PFD) and more importantly the physiological traits underlying genotypic variation. The objectives of this study were to determine the effects of moisture stress during the pre-flowering phase on pod yield and to understand some of the physiological responses underlying genotypic variation in response to and recovery from PFD. A glasshouse and field experiments were conducted at Khon Kaen University, Thailand. The glasshouse experiment was a randomized complete block design consisting of two watering regimes, i.e. fully-irrigated control and 1/3 available soil water from emergence to 40 days after emergence followed by adequate water supply, and 12 peanut genotypes. The field experiment was a split-plot design with two watering regimes as main-plots, and 12 peanut genotypes as sub-plots. Measurements of N-2 fixation, leaf area (LA) were made in both experiments. In addition, root growth was measured in the glasshouse experiment. Imposition of PFD followed by recovery resulted in an average increase in yield of 24 % (range from 10 % to 57 %) and 12 % (range from 2 % to 51 %) in the field and glasshouse experiments, respectively. Significant genotypic variation for N-2 fixation, LA and root growth was also observed after recovery. The study revealed that recovery growth following release of PFD had a stronger influence on final yield than tolerance to water deficits during the PFD. A combination of N-2 fixation, LA and root growth accounted for a major portion of the genotypic variation in yield (r = 0.68-0.93) suggesting that these traits could be used as selection criteria for identifying genotypes with rapid recovery from PFD. A combined analysis of glasshouse and field experiments showed that LA and N-2 fixation during the recovery had low genotype x environment interaction indicating potential for using these traits for selecting genotypes in peanut improvement programs.

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.