Enzymatic activity other than lipase

In this chapter, enzymes (other than lipase) which are present in cream are discussed. The effects of heat treatments on the activities of these enzymes are described. The influence of residual enzyme activiv, remaining after heating, on cream quality is also discussed.

Epithiospecifier protein activity in broccoli: The link between terminal alkenyl glucosinolates and sulphoraphane nitrile

The chemical nature of the hydrolysis products from the glucosinolate-myrosinase system depends on the presence or absence of supplementary proteins, such as epithiospecifier proteins (ESPs). ESPs (non-catalytic cofactors of myrosinase) promote the formation of epithionitriles from terminal alkenyl glucosinolates and as recent evidence suggests, simple nitriles at the expense of isothiocyanates. The ratio of ESP activity to myrosinase activity is crucial in determining the proportion of these nitriles produced on hydrolysis. Sulphoraphane, a major isothiocyanate produced in broccoli seedlings, has been found to be a potent inducer of phase 2 detoxification enzymes. However, ESP may also support the formation of the non-inductive sulphoraphane nitrile. Our objective was to monitor changes in ESP activity during the development of broccoli seedlings and link these activity changes with myrosinase activity, the level of terminal alkenyl glucosinolates and sulphoraphane nitrile formed. Here, for the first time, we show ESP activity increases up to day 2 after germination before decreasing again to seed activity levels at day 5. These activity changes paralleled changes in myrosinase activity and terminal alkenyl glucosinolate content. There is a significant relationship between ESP activity and the formation of sulforaphane nitrile in broccoli seedlings. The significance of these findings for the health benefits conferred by eating broccoli seedlings is briefly discussed.

Models for the field-based toxicity of copper and zinc salts to wheat in 11 Australian soils and comparison to laboratory-based models

Laboratory-based relationships that model the phytotoxicity of metals using soil properties have been developed. This paper presents the first field-based phytotoxicity relationships. Wheat(Triticum aestivum L) was grown at 11 Australian field sites at which soil was spiked with copper (Cu) and zinc (Zn) salts. Toxicity was measured as inhibition of plant growth at 8 weeks and grain yield at harvest. The added Cu and Zn EC10 values for both endpoints ranged from approximately 3 to 4760 mg/kg. There were no relationships between field-based 8-week biomass and grain yield toxicity values for either metal. Cu toxicity was best modelled using pH and organic carbon content while Zn toxicity was best modelled using pH and the cation exchange capacity. The best relationships estimated toxicity within a factor of two of measured values. Laboratory-based phytotoxicity relationships could not accurately predict field-based phytotoxicity responses.

Bioavailability of zinc and copper in biosolids compared to their soluble salts.

For essential elements, such as copper (Cu) and zinc (Zn), the bioavailability in biosolids is important from a nutrient release and a potential contamination perspective. Most ecotoxicity studies are done using metal salts and it has been argued that the bioavailability of metals in biosolids can be different to that of metal salts. We compared the bioavailability of Cu and Zn in biosolids with those of metal salts in the same soils using twelve Australian field trials. Three different measures of bioavailability were assessed: soil solution extraction, CaCl2 extractable fractions and plant uptake. The results showed that bioavailability for Zn was similar in biosolid and salt treatments. For Cu, the results were inconclusive due to strong Cu homeostasis in plants and dissolved organic matter interference in extractable measures. We therefore recommend using isotope dilution methods to assess differences in Cu availability between biosolid and salt treatments.

An innovative microplate assay to facilitate the detection of antimicrobial activity in plant extracts.

A microplate assay was modified for the detection of antimicrobial activity in plant extracts. The aim was to develop an in vitro assay that could rapidly screen plant extracts to provide quantitative data on inhibition of microbial growth. A spectrophotometric assay using a microplate with serial dilutions of the plant extract and the bacteria was developed. Two bacteria, Staphylococcus aureus and Escherichia coli, were used for this study. Essential oils, oregano (Origanum vulgare) and lemon myrtle (Backhousia citriodora), and three active components carvacrol, thymol and citral were evaluated. The reproducibility of the assay was high, with correlation coefficients (r aureus and E. coli between 0.9321 and 0.9816. Similarly, r and 0.9814. This assay could also be used to measure antimicrobial activity in plant extracts which vary in pH and color.

Key Role of Fe2+ in Epithiospecifier Protein Activity.

The chemical nature of the hydrolysis products from the glucosinolate-myrosinase system depends on the presence or absence of supplementary proteins such as epithiospecifier proteins (ESPs). ESPs promote the formation of epithionitriles from terminal alkenyl glucosinolates and, as recent evidence suggests, simple nitriles at the expense of isothiocyanates. From a human health perspective isothiocyanates are the most important because they are major inducers of carcinogen-detoxifying enzymes. Fe2+ is an essential factor in ESP activity, although several recent studies have highlighted discrepancies in the understanding of the ESP-iron interaction. To investigate further the role iron species play in regulating ESP activity, four ESP-containing seedpowders were analyzed for ESP and myrosinase activities, endogenous iron content, and glucosinolate degradation products after the addition of iron species, specific chelators, and reducing agents. For the first time this paper shows the effect of these additions on the hydrolysis of individual glucosinolates that constitute the total pool. Aged seeds and 3-day seedlings were also tested to investigate the effects of seed storage and early plant development on iron levels and ESP activity. The four ESP-containing plant systems tested gave two distinctive responses, thus providing strong evidence that ESPs vary markedly in their Fe2+ requirement for activity. The results also indicated that reduction of ferric to ferrous iron drives variations in ESP activity during early plant development. The reverse oxidation reaction provided a convincing explanation for the loss of ESP activity during seed storage. Aged seeds produced seedlings with substantially lower ESP activity, and there was a concomitant loss in germination rate. It was concluded that manipulation of endogenous iron levels of ESP-containing plants could increase the conversion of glucosinolates to isothiocyanates and enhance potential health benefits.

A hidden Markov model approach for determining vessel activity from vessel monitoring system data.

Many fisheries worldwide have adopted vessel monitoring systems (VMS) for compliance purposes. An added benefit of these systems is that they collect a large amount of data on vessel locations at very fine spatial and temporal scales. This data can provide a wealth of information for stock assessment, research, and management. However, since most VMS implementations record vessel location at set time intervals with no regard to vessel activity, some methodology is required to determine which data records correspond to fishing activity. This paper describes a probabilistic approach, based on hidden Markov models (HMMs), to determine vessel activity. A HMM provides a natural framework for the problem and, by definition, models the intrinsic temporal correlation of the data. The paper describes the general approach that was developed and presents an example of this approach applied to the Queensland trawl fishery off the coast of eastern Australia. Finally, a simulation experiment is presented that compares the misallocation rates of the HMM approach with other approaches.

An exploratory study to assess the activity of the acarine growth inhibitor, fluazuron, against Sarcoptes scabei infestation in pigs

Background: The most common treatments for scabies in human and veterinary settings are topical 5% permethrin or systemic treatment with ivermectin. However, these treatments have very little activity against arthropod eggs, and therefore repeated treatment is frequently required. In-vitro, biochemical and molecular studies have demonstrated that human mites are becoming increasingly resistant to both acaricides. To identify alternate acaricides, we undertook a pilot study of the in vivo activity of the benzoylphenyl urea inhibitor of chitin synthesis, fluazuron, in pigs with sarcoptic mange. Findings: Pigs (n = 5) were infested with S. scabei var suis, and randomised to treatment at the start of peak infestation with fluazuron at a dose of 10 mg/kg/day per os for 7 days (n = 3) or no treatment (n = 2). Clinical scores, skin scrapings for mite counts and blood sampling for pharmacokinetic analysis were undertaken. Fluazuron was well absorbed in treated pigs with measureable blood levels up to 4 weeks post treatment. No adverse effects were observed. Modest acaricidal activity of the compound was observed, with a reduction in severity of skin lesions in treated pigs, as well as a reduction in number of scabies mite's early life stages. Conclusions: The moderate efficacy of fluazuron against scabies mites indicates a lead to the development of alternate treatments for scabies, such as combination therapies that maybe applicable for human use in the future.

Field inoculation with arbuscular-mycorrhizal fungi overcomes phosphorus and zinc deficiencies of linseed (Linum usitatissimum) in a vertisol subject to long-fallow disorder

Long-fallow disorder is expressed as exacerbated deficiencies of phosphorus (P) and/or zinc (Zn) in field crops growing after long periods of weed-free fallow. The hypothesis that arbuscular-mycorrhizal fungi (AMF) improve the P and Zn nutrition, and thereby biomass production and seed yield of linseed (Linum usitatissimum) was tested in a field experiment. A factorial combination of treatments consisting of +/- fumigation, +/- AMF inoculation with Glomus spp., +/- P and +/- Zn fertilisers was used on a long-fallowed vertisol. The use of such methods allowed an absolute comparison of plants growing with and without AMF in the field for the first time in a soil disposed to long-fallow disorder. Plant biomass, height, P and Zn concentrations and contents, boll number and final seed yield were (a) least in fumigated soil with negligible AMF colonisation of the roots, (b) low initially in long-fallow soil but increased with time as AMF colonisation of the roots developed, and (c) greatest in soil inoculated with AMF cultures. The results showed for the first time in the field that inflows of both P and Zn into linseed roots were highly dependent on %AMF-colonisation (R-2 = 0.95 for P and 0.85 for Zn, P < 0.001) in a soil disposed to long-fallow disorder. Relative field mycorrhizal dependencies without and with P+Zn fertiliser were 85 % and 86 % for biomass and 68 % and 52 % for seed yield respectively. This research showed in the field that AMF greatly improved the P and Zn nutrition, biomass production and seed yield of linseed growing in a soil disposed to long-fallow disorder. The level of mycorrhizal colonisation of plants suffering from long-fallow disorder can increase during the growing season resulting in improved plant growth and residual AMF inoculum in the soil, and thus it is important for growers to recognise the cause and not terminate a poor crop prematurely in order to sow another. Other positive management options to reduce long fallows and foster AMF include adoption of conservation tillage and opportunity cropping.

Alk (en) ylresorcinol concentrations in'Kensington Pride'mango peel and antifungal activity against Colletotrichum gloeosporioides

Two preformed alk(en)ylresorcinols, 5-n-heptadecenylresorcinol and 5-n-pentadecylresorcinol, were identified in ‘Kensington Pride’ mango fruit peel. The alk(en)ylresorcinols had antifungal activity against C. gloeosporioides, as determined from thin layer chromatography bioassays. Soil-applied activators of plant defence (Acibenzolar at 150 mg L-1, and soluble potassium silicate at 200 and 1000 mg L-1) did not influence concentrations of 5-n-heptadecenylresorcinol or 5-n-pentadecyl¬resorcinol in mango peel when applied 2 months after fruit set and one month later. Concentrations of both alk(en)ylresorcinols were high 2 months after fruit set but levels declined by 50% within 1 month (2 months before commercial harvest) and did not change significantly from commercial harvest until eating-ripe.

Infrared spectroscopy as a rapid tool to detect methylglyoxal and antibacterial activity in Australian honeys

Methylglyoxal (2-oxopropanal) is a compound known to contribute to the non-peroxide antimicrobial activity of honeys. The feasibility of using infrared spectroscopy as a predictive tool for honey antibacterial activity and methylglyoxal content was assessed. A linear relationship was found between methylglyoxal content (279–1755 mg/kg) in Leptospermum polygalifolium honeys and bacterial inhibition for Escherichiacoli (R2 = 0.80) and Staphylococcusaureus (R2 = 0.64). A good prediction of methylglyoxal (R2 0.75) content in honey was achieved using spectroscopic data from the mid infrared (MIR) range in combination with partial least squares regression. These results indicate that robust predictive equations could be developed using MIR for commercial application where the prediction of bacterial inhibition is needed to ‘value’ honeys with methylglyoxal contents in excess of 200 mg/kg.

Using UHF proximity loggers to quantify male–female interactions: A scoping study of estrous activity in cattle

Reproductive efficiency is an important determinant of profitable cattle breeding systems and the success of assisted reproductive techniques (ART) in wildlife conservation programs. Methods of estrous detection used in intensive beef and dairy cattle systems lack accuracy and remain the single biggest issue for improvement of reproductive rates and such methods are not practical for either large-scale extensive beef cattle enterprises or free-living mammalian species. Recent developments in UHF (ultra high frequency) proximity logger telemetry devices have been used to provide a continuous pair-wise measure of associations between individual animals for both livestock and wildlife. The objective of this study was to explore the potential of using UHF telemetry to identify the reproductive cycle phenotype in terms of intensity and duration of estrus. The study was conducted using Belmont Red (interbred Africander Brahman Hereford–Shorthorn) cattle grazing irrigated pasture on Belmont Research Station, northeastern Australia. The cow-bull associations from three groups of cows each with one bull were recorded over a 7-week breeding season and the stage of estrus was identified using ultrasonography. Telemetry data from bull and cows, collected over 4 8-day logger deployments, were log transformed and analyzed by ANOVA. Both the number and duration of bull-cow affiliations were significantly (P < 0.001) greater in estrous cows compared to anestrus cows. These results support the development of the UHF technology as a hands-off and noninvasive means of gathering socio-sexual information on both wildlife and livestock for reproductive management.

Electrophysiological activity of the Sirex noctilio ovipositor: You know the drill?

Sirex woodwasp (Sirex noctilio F: Hymenoptera: Siricidae) is a major worldwide pest of pine (Pinus) species. The female woodwasp undertakes exploratory drills with the ovipositor prior to egg-laying to assess tree suitability. Previous work has shown that this behaviour is associated with assessing the osmotic pressure of the tree. Here we show that, in addition, the ovipositor is electrophysiologically active and capable of detecting ethanol and chemical components of solvent extracts of pine needles and bark. Scanning electron micrographs of the ovipositor show the presence of structures which may have a chemoreceptive function. Our research expands our knowledge of the role that the Sirex ovipositor plays in egg-laying site selection.

Cytorhabdovirus phosphoprotein shows RNA silencing suppressor activity in plants, but not in insect cells

RNA silencing in plants and insects provides an antiviral defense and as a countermeasure most viruses encode RNA silencing suppressors (RSS). For the family Rhabdoviridae, no detailed functional RSS studies have been reported in plant hosts and insect vectors. In agroinfiltrated Nicotiana benthamiana leaves we show for the first time for a cytorhabdovirus, lettuce necrotic yellows virus (LNYV), that one of the nucleocapsid core proteins, phosphoprotein (P) has relatively weak local RSS activity and delays systemic silencing of a GFP reporter. Analysis of GFP small RNAs indicated that the P protein did not prevent siRNA accumulation. To explore RSS activity in insects, we used a Flock House virus replicon system in Drosophila S2 cells. In contrast to the plant host, LNYV P protein did not exhibit RSS activity in the insect cells. Taken together our results suggest that P protein may target plant-specific components of RNA silencing post siRNA biogenesis.

Water activity of poultry litter: Relationship to moisture content during a grow-out

Poultry grown on litter floors are in contact with their own waste products. The waste material needs to be carefully managed to reduce food safety risks and to provide conditions that are comfortable and safe for the birds. Water activity (Aw) is an important thermodynamic property that has been shown to be more closely related to microbial, chemical and physical properties of natural products than moisture content. In poultry litter, Aw is relevant for understanding microbial activity; litter handling and rheological properties; and relationships between in-shed relative humidity and litter moisture content. We measured the Aw of poultry litter collected throughout a meat chicken grow-out (from fresh pine shavings bedding material to day 52) and over a range of litter moisture content (10–60%). The Aw increased non-linearly from 0.71 to 1.0, and reached a value of 0.95 when litter moisture content was only 22–33%. Accumulation of manure during the grow-out reduced Aw for the same moisture content. These results are relevant for making decisions regarding litter re-use in multiple grow-outs as well as setting targets for litter moisture content to minimise odour, microbial risks and to ensure necessary litter physical conditions are maintained during a grow-out. Methods to predict Aw in poultry litter from moisture content are proposed.