The effect of post harvest handling on selected native food plants

A commercial issue currently facing native plant food producers and food processors, and identified by the industry itself, is that of delivering quality products consistently and at reasonable cost to end users based on a sound food technology and nutrition platform. A literature survey carried out in July 2001 by the DPI&F’s Centre for Food Technology, Brisbane in collaboration with the University of Queensland to collect the latest information at that time on the functional food market as it pertained to native food plants, indicated that little or no work had been published on this topic. This project addresses two key RIRDC sub program strategies: to identify and evaluate processes or products with prospects of commercial viability and to assist in the development of integrated production, harvesting, processing and marketing systems. This project proposal also reflects a key RIRDC R&D issue for 2002-2003; that of linking with prospective members of the value chain. The purpose of this project was to obtain chemical data on the post harvest stability of functional nutritional components (bio actives) in commercially available, hand harvested bush tomato and Kakadu plum. The project concentrated on evaluating bioactive stability as a measure of ingredient quality.

Handling of Two Tropical Australian Sharks to Improve Quality and to Identify the Cause of Tough Texture

Sharks caught in tropical Australian waters occasionally exhibit tough texture. Two species of Carcharinid shark, originally known as the sorrah shark (Carcharinus sorrah) and the black spot shark (Carcharinus tilstoni), compose the majority of the catch. Experiments were conducted to identify the cause of tough texture and to improve the overall quality of the catch. The possibility that a cold shock reaction may occur was investigated by observing the contraction of fillets under a range of temperature conditions before freezing. The effect of on-board handling practices were evaluated using frozen shark fillets, which had been stored prior to freezing in refrigerated seawater at different rigor stages, temperatures and times as trunks. Fillets were analyzed for nucleotides, lactate, thaw pH, sarcomere length and raw and cooked shear force values. The existence of thaw rigor was also investigated. There was little difference in the texture between the individual strips of a fillet exposed to different temperatures but there were significant differences between individual sharks. A cold shock reaction could not be demonstrated in these species. The main influences on texture were of biological origin. The species, sex and size were found to have significant links with texture of fillets. The quality of the fillets deteriorated quicker during the warmer season and were at their worst if the trunks were kept on deck till post-rigor or held in 15 degree C refrigerated seawater before freezing

Effects of Nutrition and Time of Weaning on Dry Season Liveweight Loss of Breeder Cows

In the seasonally dry tropics of northern Australia, breeder cows may lose up to 30% liveweight during the dry season when pasture is of low nutritive value. This is a major cause of low reproductive rates and high mortality. Weaning early in the dry season is effective to reduce this liveweight loss of the breeder (Holroyd et al. 1988). An experiment examined the dry season liveweight loss of breeders for a range of weaning times and levels of nutrition. From April to October through the dry season, 209 Bos indicus x Shorthorn cross cows 4-6 years of age grazed speargrass pastures in north Queensland. The cows had been joined with bulls from late January until April. Twenty-nine breeders had not suckled a calf during the previous wet season (DRY cows). In addition 180 cows lactating in April were weaned in late April, mid July or early September. The cows were allocated by stratified randomisation based on lactational status, stage of pregnancy and body condition to 15 x 40 ha paddocks. Five paddocks with low fertility soils provided LOW nutrition, while 10 paddocks with medium fertility soils and no supplementation or with supplementation provided MEDIUM and HIGH nutrition, respectively. The supplement consisted of molasses containing 14% urea offered ad libitum. Liveweight was measured at intervals and conceptus-free liveweight (CF-LW) calculated. Data were analyses by AOV within groups of paddocks. Animal production for a consuming world : proceedings of 9th Congress of the Asian-Australasian Association of Animal Production Societies [AAAP] and 23rd Biennial Conference of the Australian Society of Animal Production [ASAP] and 17th Annual Symposium of the University of Sydney, Dairy Research Foundation, [DRF]. 2-7 July 2000, Sydney, Australia.

Timing of weaning and processing: effects on liveweight changes of weaner cattle

To reduce the number of musters and handling costs, calves in extensive cattle herds in northern Australia are processed (vaccinated, ear-marked, de-horned, branded and males castrated) shortly after they are weaned. As stress has adverse effects on health and growth, and weaning is a stressful time for calves, this experiment asked if calf health, welfare and performance were improved when calves had a period with their mothers post-processing, before they were weaned.

Predicting survival of western rock lobsters Panulirus cygnus using discriminant analysis of hemolymph parameters taken immediately following simulated handling treatments

Instances of morbidity amongst rock lobsters (Panulirus cygnus) arriving at factories in Western Australia (WA) have been attributed to stress during post-harvest handling. This study used discriminant analysis to determine whether physiological correlates of stress following a period of simulated post-harvest handling had any validity as predictors of future rejection or morbidity of western rock lobsters. Groups of 230 western rock lobsters were stored for 6 h in five environments (submerged/flowing sea water, submerged/re-circulating sea water, humid air, flowing sea water spray, and re-circulated sea water spray). The experiment was conducted in late spring (ambient sea water 22°C), and repeated again in early autumn (ambient sea water 26°C). After 6 h treatment, each lobster was graded for acceptability for live export, numbered, and its hemolymph was sampled. The samples were analysed for a number of physiological and health status parameters. The lobsters were then stored for a week in tanks in the live lobster factory to record mortality. The mortality of lobsters in the factory was associated with earlier deviations in hemolymph parameters as they emerged from the storage treatments. Discriminant analysis (DA) of the hemolymph assays enabled the fate of 80-90% of the lobsters to be correctly categorised within each experiment. However, functions derived from one experiment were less accurate at predicting mortality when applied to the other experiments. One of the reasons for this was the higher mortality and the more severe patho-physiological changes observed in lobsters stored in humid air or sprays at the higher temperature. The analysis identified lactate accumulation during emersion and associated physiological and hemocyte-related effects as a major correlate of mortality. Reducing these deviations, for example by submerged transport, is expected to ensure high levels of survival. None of the indicators tested predicted mortality with total accuracy. The simplest and most accurate means of comparing emersed treatments was to count the mortality afterwards.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy® or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to 1 infected in 800 samples with pepper but never detecting more than 1 infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Assessing the relationship between patch type and soil mites: A molecular approach

An urgent need exists for indicators of soil health and patch functionality in extensive rangelands that can be measured efficiently and at low cost. Soil mites are candidate indicators, but their identification and handling is so specialised and time-consuming that their inclusion in routine monitoring is unlikely. The aim of this study was to measure the relationship between patch type and mite assemblages using a conventional approach. An additional aim was to determine if a molecular approach traditionally used for soil microbes could be adapted for soil mites to overcome some of the bottlenecks associated with soil fauna diversity assessment. Soil mite species abundance and diversity were measured using conventional ecological methods in soil from patches with perennial grass and litter cover (PGL), and compared to soil from bare patches with annual grasses and/or litter cover (BAL). Soil mite assemblages were also assessed using a molecular method called terminal-restriction fragment length polymorphism (T-RFLP) analysis. The conventional data showed a relationship between patch type and mite assemblage. The Prostigmata and Oribatida were well represented in the PGL sites, particularly the Aphelacaridae (Oribatida). For T-RFLP analysis, the mite community was represented by a series of DNA fragment lengths that reflected mite sequence diversity. The T-RFLP data showed a distinct difference in the mite assemblage between the patch types. Where possible, T-RFLP peaks were matched to mite families using a reference 18S rDNA database, and the Aphelacaridae prevalent in the conventional samples at PGL sites were identified, as were prostigmatids and oribatids. We identified limits to the T-RFLP approach and this included an inability to distinguish some species whose DNA sequences were similar. Despite these limitations, the data still showed a clear difference between sites, and the molecular taxonomic inferences also compared well with the conventional ecological data. The results from this study indicated that the T-RFLP approach was effective in measuring mite assemblages in this system. The power of this technique lies in the fact that species diversity and abundance data can be obtained quickly because of the time taken to process hundreds of samples, from soil DNA extraction to data output on the gene analyser, can be as little as 4 days.

Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction.

Equid herpesvirus 1 (EHV1) is a major disease of equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-binding probe (MGB) technology for the diagnosis of EHV1. This technique does not require post-PCR manipulations thereby reducing the risk of cross-contamination. Most importantly, the technique is specific; it was able to differentiate EHV1 from the closely related member of the Alphaherpesvirinae, equid herpesvirus 4 (EHV4). It was not reactive with common opportunistic pathogens such as Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Enterobacter agglomerans often involved in abortion. Similarly, it did not react with equine pathogens such as Streptococcus equi, Streptococcus equisimilis, Streptococcus zooepidemicus, Taylorella equigenitalis and Rhodococcus equi, which also cause abortion. The results obtained with this technique agreed with results from published PCR methods. The assay was sensitive enough to detect EHV1 sequences in paraffin-embedded tissues and clinical samples. When compared to virus isolation, the test was more sensitive. This test will be useful for the routine diagnosis of EHV1 based on its specificity, sensitivity, ease of performance and rapidity.

Multiplex real-time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4).

A multiplex real-time PCR was designed to detect and differentiate equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB?) labelled with 6-carboxy-fluorescein (FAM?) and VIC® for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaherpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.

How effective are broad-scale nutrient mass balances for determining the sustainability of lot-feed manure application?

Nutrient mass balances have been used to assess a variety of land resource scenarios, at various scales. They are widely used as a simple basis for policy, planning, and regulatory decisions but it is not clear how accurately they reflect reality. This study provides a critique of broad-scale nutrient mass balances, with particular application to the fertiliser use of beef lot-feeding manure in Queensland. Mass balances completed at the district and farm scale were found to misrepresent actual manure management behaviour and potentially the risk of nutrient contamination of water resources. The difficulties of handling stockpile manure and concerns about soil compaction mean that manure is spread thickly over a few paddocks at a time and not evenly across a whole farm. Consequently, higher nutrient loads were applied to a single paddock less frequently than annually. This resulted in years with excess nitrogen, phosphorus, and potassium remaining in the soil profile. This conclusion was supported by evidence of significant nutrient movement in several of the soil profiles studied. Spreading manure is profitable, but maximum returns can be associated with increased risk of nutrient leaching relative to conventional inorganic fertiliser practices. Bio-economic simulations found this increased risk where manure was applied to supply crop nitrogen requirements (the practice of the case study farms, 200-5000 head lot-feeders). Thus, the use of broad-scale mass balances can be misleading because paddock management is spatially heterogeneous and this leads to increased local potential for nutrient loss. In response to the effect of spatial heterogeneity policy makers who intend to use mass balance techniques to estimate potential for nutrient contamination should apply these techniques conservatively.

Value-adding Australian sardines: Factors affecting rates of deterioration in sardine (Sardinops sagax) quality during post-harvest handling

Deficiencies in sardine post-harvest handling methods were seen as major impediments to development of a value-adding sector supplying Australian bait and human consumption markets. Factors affecting sardine deterioration rates in the immediate post-harvest period were investigated and recommendations made for alternative handling procedures to optimise sardine quality. Net to factory sampling showed that post-mortem autolysis was probably caused by digestive enzyme activity contributing to the observed temporal increase in sardine Quality Index. Belly burst was not an issue. Sardine quality could be maintained by reducing tank loading, and rapid temperature reduction using dedicated, on-board value-adding tanks. Fish should be iced between the jetty and the processing factory, and transport bins chilled using an efficient cooling medium such as flow ice.

Responses in carcass lean pH at 24 hours post-mortem in lines of Large White pigs selected for growth rate on a fixed ration over a set time

A restricted maximum likelihood analysis applied to an animal model showed no significant differences (P > 0.05) in pH value of the longissimus dorsi measured at 24 h post-mortem (pH24) between high and low lines of Large White pigs selected over 4 years for post-weaning growth rate on restricted feeding. Genetic and phenotypic correlations between pH24 and production and carcass traits were estimated using all performance testing records combined with the pH24 measurements (5.05-7.02) on slaughtered animals. The estimate of heritability for pH24 was moderate (0.29 ± 0.18). Genetic correlations between pH24 and production or carcass composition traits, except for ultrasonic backfat (UBF), were not significantly different from zero. UBF had a moderate, positive genetic correlation with pH24 (0.24 ± 0.33). These estimates of genetic correlations affirmed that selection for increased growth rate on restricted feeding is likely to result in limited changes in pH24 and pork quality since the selection does not put a high emphasis on reduced fatness.

Time-dependent instantaneous mortality rates from multiple tagging experiments with exact times of release and recovery

Instantaneous natural mortality rates and a nonparametric hunting mortality function are estimated from a multiple-year tagging experiment with arbitrary, time-dependent fishing or hunting mortality. Our theory allows animals to be tagged over a range of times in each year, and to take time to mix into the population. Animals are recovered by hunting or fishing, and death events from natural causes occur but are not observed. We combine a long-standing approach based on yearly totals, described by Brownie et al. (1985, Statistical Inference from Band Recovery Data: A Handbook, Second edition, United States Fish and Wildlife Service, Washington, Resource Publication, 156), with an exact-time-of-recovery approach originated by Hearn, Sandland and Hampton (1987, Journal du Conseil International pour l'Exploration de la Mer, 43, 107-117), who modeled times at liberty without regard to time of tagging. Our model allows for exact times of release and recovery, incomplete reporting of recoveries, and potential tag shedding. We apply our methods to data on the heavily exploited southern bluefin tuna (Thunnus maccoyii).

Absolute quantitation of Marek's disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR

The further development of Taqman quantitative real-time PCR (qPCR) assays for the absolute quantitation of Marek's disease virus serotype 1 (MDV1) and Herpesvirus of turkeys (HVT) viruses is described and the sensitivity and reproducibility of each assay reported. Using plasmid DNA copies, the lower limit of detection was determined to be 5 copies for the MDV1 assay and 75 copies for the HVT assay. Both assays were found to be highly reproducible for Ct values and calculated copy numbers with mean intra- and inter-assay coefficients of variation being less than 5% for Ct and 20% for calculated copy number. The genome copy number of MDV1 and HVT viruses was quantified in PBL and feather tips from experimentally infected chickens, and field poultry dust samples. Parallelism was demonstrated between the plasmid-based standard curves, and standard curves derived from infected spleen material containing both viral and host DNA, allowing the latter to be used for absolute quantification. These methods should prove useful for the reliable differentiation and absolute quantitation of MDV1 and HVT viruses in a wide range of samples.

Testing the simulation capability of APSIM-ORYZA under different levels of nitrogen fertiliser and transplanting time regimes in Korea

APSIM-ORYZA is a new functionality developed in the APSIM framework to simulate rice production while addressing management issues such as fertilisation and transplanting, which are particularly important in Korean agriculture. To validate the model for Korean rice varieties and field conditions, the measured yields and flowering times from three field experiments conducted by the Gyeonggi Agricultural Research and Extension Services (GARES) in Korea were compared against the simulated outputs for different management practices and rice varieties. Simulated yields of early-, mid- and mid-to-late-maturing varieties of rice grown in a continuous rice cropping system from 1997 to 2004 showed close agreement with the measured data. Similar results were also found for yields simulated under seven levels of nitrogen application. When different transplanting times were modelled, simulated flowering times ranged from within 3 days of the measured values for the early-maturing varieties, to up to 9 days after the measured dates for the mid- and especially mid-to-late-maturing varieties. This was associated with highly variable simulated yields which correlated poorly with the measured data. This suggests the need to accurately calibrate the photoperiod sensitivity parameters of the model for the photoperiod-sensitive rice varieties in Korea.