Comparative studies on the invasion of cattle ticks (Rhipicephalus (Boophilus) microplus) and sheep blowflies (Lucilia cuprina) by Metarhizium anisopliae (Sorokin).

Microscopic investigations over time were carried out to study and compare the pathogenesis of invasion of ticks and blowflies by Metarhizium anisopliae. The scanning electron microscope and stereo light microscope were used to observe and record processes on the arthropods' surfaces and the compound light microscope was used to observe and record processes within the body cavities. Two distinctly different patterns of invasion were found in ticks and blowflies. Fungal conidia germinated on the surface of ticks then hyphae simultaneously penetrated into the tick body and grew across the tick surface. There was extensive fungal degradation of the tick cuticle, particularly the outer endocuticle. Although large numbers of conidia adhered to the surface of blowflies, no conidia were seen to germinate on external surfaces. A single germinating conidium was seen in the entrance to the buccal cavity. Investigations of the fly interior revealed a higher density of hyphal bodies in the haemolymph surrounding the buccal cavity than in haemolymph from regions of the upper thorax. This pattern suggests that fungal invasion of the blowfly is primarily through the buccal cavity. Plentiful extracellular mucilage was seen around the hyphae on tick cuticles, and crystals of calcium oxalate were seen amongst the hyphae on the surface of ticks and in the haemolymph of blowflies killed by M. anisopliae isolate ARIM16.

Comparative effects of thermal and high pressure processing on phenolic phytochemicals in different strawberry cultivars

Abstract It is widely considered that high pressure processing (HPP) results in better retention of micronutrients and phytochemicals compared to thermal pasteurization (TP), although some studies indicate that this may not be true in all cases. The aims of this study were (1) to objectively compare the effects of HPP under commercial processing conditions with thermal pasteurization (TP) on the stability of phenolic antioxidants in strawberries following processing and during storage and (2) to evaluate the influence of varietal differences and hence differences in biochemical composition of strawberries on the stability of phenolic antioxidants. Strawberry puree samples from cultivars Camarosa, Rubygem, and Festival were subjected to HPP (600 MPa/20 °C/5 min) and TP (88 °C/2 min). The activities of oxidative enzymes were evaluated before and after processing. Furthermore, the antioxidant capacity (total phenolic content (TPC), oxygen radical absorbance capacity (ORAC), and ferric reducing antioxidant power (FRAP)) and individual anthocyanins (by HPLC) were determined prior to and following processing and after three months of refrigerated storage (4 °C). Depending on the cultivar, HPP caused 15–38% and 20–33% inactivation of polyphenol oxidase and peroxidase, respectively, compared to almost complete inactivation of these enzymes by TP. Significant decreases (p < 0.05) in ORAC, FRAP, TPC and anthocyanin contents were observed during processing and storage of both HPP and TP samples. Anthocyanins were the most affected with only 19–25% retention after three months of refrigerated storage (4 °C). Slightly higher (p < 0.05) loss of TPC and antioxidant capacity were observed during storage of HPP samples compared to TP. Industrial Relevance: The results of the study demonstrated that both high pressure processing and thermal pasteurization result in high retention of phenolic phytochemicals in strawberry products. Under the conditions investigated, high pressure processing did not result in a better retention of phenolic phytochemicals compared to thermal pasteurization. In fact, a slightly higher loss of total polyphenol content and antioxidant capacity were observed during refrigerated storage of HPP processed samples. Our results showed that, high pressure processing may not always be a better alternative to thermal processing for strawberry puree processing if the main objective is better retention of phenolic antioxidants. However, it should be noted that other quality attributes such as sensory properties, where distinct advantages of HPP are expected, were outside the scope of this study.

Maturation studies of pecan nuts grown in Queensland

Changes in chemical composition, physical and sensory characteristics were followed in two pecan cultivars Wichita and Western Schley harvested from a commercial orchard at Gatton in Queensland seven times during 1996.Testa colour of both pecan cultivars darkened and opalescence decreased as the nuts matured. Bitterness of Western Schley pecans decreased with maturity. Colour of shuck, shell and kernel of both cultivars developed as the nuts matured. Wichita pecans were larger than Western Schley at all harvest times. Both nut-in-shell and kernel moisture decreased with maturity, whereas oil and sucrose contents increased. Both pecan cultivars had reached advanced maturation by the first harvest on March 18.

A precision apparatus, with solid phase micro-extraction monitoring capability, for incorporation studies of gaseous precursors into insect-derived metabolites

An apparatus is described that facilitates the determination of incorporation levels of isotope labelled, gaseous precursors into volatile insect-derived metabolites. Atmospheres of varying gas compositions can be generated by evacuation of a working chamber followed by admission of the required levels of component gases, using a precision, digitised pressure read-out system. Insects such as fruit-flies are located initially in a small introduction chamber, from which migration can occur downwards into the working chamber. The level of incorporation of labelled precursors is continuously assayed by the Solid Phase Micro Extraction (SPME) technique and GC-MS analyses. Experiments with both Bactrocera species (fruit-flies) and a parasitoid wasp, Megarhyssa nortoni nortoni (Cresson) and oxygen-18 labelled dioxygen illustrate the utility of this system. The isotope effects of oxygen-18 on the carbon-13 NMR spectra of 1,7- dioxaspiro[5,5]undecane are also described.

Comparative immunogenicity of M. hyopneumoniae NrdF encoded in different expression systems delivered orally via attenuated S. typhimurium aroA in mice.

The Mycoplasma hyopneumoniae ribonucleotide reductase R2 subunit (NrdF) gene fragment was cloned into eukaryotic and prokaryotic expression vectors and its immunogenicity evaluated in mice immunized orally with attenuated Salmonella typhimurium aroA CS332 harboring either of the recombinant expression plasmids. We found that NrdF is highly conserved among M. hyopneumoniae strains. The immunogenicity of NrdF was examined by analyzing antibody responses in sera and lung washes, and the cell-mediated immune (CMI) response was assessed by determining the INF-[gamma] level produced by splenocytes upon in vitro stimulation with NrdF antigen. S. typhimurium expressing NrdF encoded by the prokaryotic expression plasmid (pTrcNrdF) failed to elicit an NrdF-specific serum or secretory antibody response, and IFN-[gamma] was not produced. Similarly, S. typhimurium carrying the eukaryotic recombinant plasmid encoding NrdF (pcNrdF) did not induce a serum or secretory antibody response, but did elicit significant NrdF-specific IFN-[gamma] production, indicating induction of a CMI response. However, analysis of immune responses against the live vector S. typhimurium aroA CS332 showed a serum IgG response but no mucosal IgA response in spite of its efficient invasiveness in vitro. In the present study we show that the DNA vaccine encoding the M. hyopneumoniae antigen delivered orally via a live attenuated S. typhimurium aroA can induce a cell-mediated immune response. We also indicate that different live bacterial vaccine carriers may have an influence on the type of the immune response induced.

Process studies of odour emissions from effluent ponds using machine-based odour measurement

Replicable experimental studies using a novel experimental facility and a machine-based odour quantification technique were conducted to demonstrate the relationship between odour emission rates and pond loading rates. The odour quantification technique consisted of an electronic nose, AromaScan A32S, and an artificial neural network. Odour concentrations determined by olfactometry were used along with the AromaScan responses to train the artificial neural network. The trained network was able to predict the odour emission rates for the test data with a correlation coefficient of 0.98. Time averaged odour emission rates predicted by the machine-based odour quantification technique, were strongly correlated with volatile solids loading rate, demonstrating the increased magnitude of emissions from a heavily loaded effluent pond. However, it was not possible to obtain the same relationship between volatile solids loading rates and odour emission rates from the individual data. It is concluded that taking a limited number of odour samples over a short period is unlikely to provide a representative rate of odour emissions from an effluent pond. A continuous odour monitoring instrument will be required for that more demanding task.

Comparative productivity of irrigated short-term ryegrass (Lolium multiflorum) pasture receiving nitrogen, grown alone or in a mixture with white (Trifolium repens) and Persian (T. resupinatum) clovers

Dairy farms in subtropical Australia use irrigated, annually sown short-term ryegrass (Lolium multiflorum) or mixtures of short-term ryegrass and white (Trifolium repens) and Persian (shaftal) (T. resupinatum) clover during the winter-spring period in all-year-round milk production systems. A series of small plot cutting experiments was conducted in 3 dairying regions (tropical upland, north Queensland, and subtropical southeast Queensland and northern New South Wales) to determine the most effective rate and frequency of application of nitrogen (N) fertiliser. The experiments were not grazed, nor was harvested material returned to the plots, after sampling. Rates up to 100 kg N/ha.month (as urea or calcium ammonium nitrate) and up to 200 kg N/ha every 2 months (as urea) were applied to pure stands of ryegrass in 1991. In 1993 and 1994, urea, at rates up to 150 kg N/ha.month and to 200 kg N/ha every 2 months, was applied to pure stands of ryegrass; urea, at rates up to 50 kg N/ha.month, was also applied to ryegrass-clover mixtures. The results indicate that applications of 50-85 kg N/ha.month can be recommended for short-term ryegrass pastures throughout the subtropics and tropical uplands of eastern Australia, irrespective of soil type. At this rate, dry matter yields will reach about 90% of their potential, forage nitrogen concentration will be increased, there is minimal risk to stock from nitrate poisoning and there will be no substantial increase in soil N. The rate of N for ryegrass-clover pastures is slightly higher than for pure ryegrass but, at these rates, the clover component will be suppressed. However, increased ryegrass yields and higher forage nitrogen concentrations will compensate for the reduced clover component. At application rates up to 100 kg N/ha.month, build-up of NO3--N and NH4+-N in soil was generally restricted to the surface layers (0-20 cm) of the soil, but there was a substantial increase throughout the soil profile at 150 kg N/ha.month. The build-up of NO3--N and NH4+-N was greater and was found at lower rates on the lighter soil compared with heavy clays. Generally, most of the soil N was in the NO3--N form and most was in the top 20 cm.

Comparative performance of Corymbia hybrids and parental species in subtropical Queensland and implications for breeding and deployment.

Eighty six full-sib Corymbia F1 hybrid families (crosses between C. torelliana and four spotted gum taxa: C. citriodora subsp. variegata, C. citriodora subsp. citriodora, C. henryi and C. maculata), were planted in six trials across six disparate sites in south-eastern Queensland to evaluate their productivity and determine their potential utility for plantation forestry. In each trial, the best-growing 20% of hybrid families grew significantly faster (P=0.05) than open-pollinated seedlots of the parent species Corymbia citriodora subsp. variegata, ranging from 107% to 181% and 127% to 287% of the height and diameter respectively. Relative performance of hybrid families growing on more than one site displayed consistency in ranking for growth across sites and analysis showed low genotype-by-environment interaction. Heritability estimates based on female and male parents across two sites at age six years for height and diameter at breast height, were high (0.62±0.28 to 0.64±0.35 and 0.31±0.21 to 0.69±0.37 respectively), and low to moderate (0.03±0.04 to 0.33±0.22) for stem straightness, branch size, incidence of ramicorns, and frost and disease resistance traits at ages one to three years. The proportion of dominance variance for height and diameter had reduced to zero by age six years. Based on these promising results, further breeding and pilot-scale family forestry and clonal forestry deployment is being undertaken. These results have also provided insights regarding the choice of a future hybrid breeding strategy.

Comparative analysis of Tritrichomonas foetus (Riedmuller, 1928) cat genotype, T. foetus (Riedmuller, 1928) cattle genotype and Tritrichomonas suis (Davaine, 1875) at 10 DNA loci

The parasitic protists in the genus Tritrichomonas cause significant disease in domestic cattle and cats. To assess the genetic diversity of feline and bovine isolates of Tritrichomonas foetus (Riedmuller, 1928) Wenrich and Emmerson, 1933, we used 10 different genetic regions, namely the protein coding genes of cysteine proteases 1,2 and 4-9 (CP1, 2, 4-9) involved in the pathogenesis of the disease caused by the parasite. The cytosolic malate dehydrogenase 1 (MDH1) and internal transcribed spacer region 2 of the rDNA unit (ITS2) were included as additional markers. The gene sequences were compared with those of Tritrichomonas suis (Davaine. 1875) Morgan and Hawkins, 1948 and Tritrichomonas mobilensis Culberson et al., 1986. The study revealed 100% identity for all 10 genes among all feline isolates (=T. foetus cat genotype), 100% identity among all bovine isolates (=T. foetus cattle genotype) and a genetic distinctness of 1% between the cat and cattle genotypes of T. foetus. The cattle genotype of T. foetus was 100% identical to T. suis at nine loci (CP1, 2,4-8, ITS2, MDH1). At CP9, three out of four T. suis isolates were identical to the T. foetus cattle genotype, while the T. suis isolate SUI-H3B sequence contained a single unique nucleotide substitution. Tritrichomonas mobilensis was 0.4% and 0.7% distinct from the cat and cattle genotypes of T. foetus, respectively. The genetic differences resulted in amino acid changes in the CP genes, most pronouncedly in CP2, potentially providing a platform for elucidation of genotype-specific host-pathogen interactions of T. foetus. On the basis of this data we judge T. suis and T. foetus to be subjective synonyms. For the first time, on objective nomenclatural grounds, the authority of T. suis is given to Davaine, 1875, rather than the commonly cited Gruby and Delafond, 1843. To maintain prevailing usage of T. foetus, we are suppressing the senior synomym T. suis Davaine, 1875 according to Article 23.9, because it has never been used as a valid name after 1899 and T. foetus is widely discussed as the cause of bovine trichomonosis. Thus bovine, feline and porcine isolates should all be given the name T. foetus. This promotes the stability of T. foetus for the veterinary and economically significant venereal parasite causing bovine trichomonosis. (C) 2012 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Eriophyid mites on spotted gums: population and histological damage studies of an emerging pest

A suite of co-occurring eriophyid mite species are significant pests in subtropical Australia, causing severe discolouration, blistering, necrosis and leaf loss to one of the region's most important hardwood species, Corymbia citriodora subsp. variegata (F. Muell.) K. D. Hill & L. A. S. Johnson (Myrtaceae). In this study, we examined mite population dynamics and leaf damage over a 1-year period in a commercial plantation of C. citriodora subsp. variegata. Our aims were to link the incidence and severity of mite damage, and mite numbers, to leaf physical traits (moisture content and specific leaf weight (SLW)); to identify any seasonal changes in leaf surface occupancy (upper vs. lower lamina); and host tree canopy strata (upper, mid or lower canopy). We compared population trends with site rainfall, temperature and humidity. We also examined physical and anatomical changes in leaf tissue in response to mite infestation to characterize the plants' physiological reaction to feeding, and how this might affect photosynthesis. Our main findings included positive correlations with leaf moisture content and mite numbers and with mite numbers and damage severity. Wet and dry leaf mass and SLW were greater for damaged tissue than undamaged tissue. Mites were distributed equally throughout the canopy and on both leaf surfaces. No relationships with climatic factors were found. Damage symptoms occurred equally and were exactly mirrored on both leaf surfaces. Mite infestation increased the overall epidermal thickness and the number and size of epidermal cells and was also associated with a rapid loss of chloroplasts from mesophyll cells beneath damage sites. The integrity of the stomatal complex was severely compromised in damaged tissues. These histological changes suggest that damage by these mites will negatively impact the photosynthetic efficiency of susceptible plantation species.

Eriophyid mites on spotted gums: population and histological damage studies of an emerging pest

A suite of co-occurring eriophyid mite species are significant pests in subtropical Australia, causing severe discolouration, blistering, necrosis and leaf loss to one of the region's most important hardwood species, Corymbia citriodora subsp. variegata (F. Muell.) K. D. Hill & L. A. S. Johnson (Myrtaceae). In this study, we examined mite population dynamics and leaf damage over a 1-year period in a commercial plantation of C. citriodora subsp. variegata. Our aims were to link the incidence and severity of mite damage, and mite numbers, to leaf physical traits (moisture content and specific leaf weight (SLW)); to identify any seasonal changes in leaf surface occupancy (upper vs. lower lamina); and host tree canopy strata (upper, mid or lower canopy). We compared population trends with site rainfall, temperature and humidity. We also examined physical and anatomical changes in leaf tissue in response to mite infestation to characterize the plants' physiological reaction to feeding, and how this might affect photosynthesis. Our main findings included positive correlations with leaf moisture content and mite numbers and with mite numbers and damage severity. Wet and dry leaf mass and SLW were greater for damaged tissue than undamaged tissue. Mites were distributed equally throughout the canopy and on both leaf surfaces. No relationships with climatic factors were found. Damage symptoms occurred equally and were exactly mirrored on both leaf surfaces. Mite infestation increased the overall epidermal thickness and the number and size of epidermal cells and was also associated with a rapid loss of chloroplasts from mesophyll cells beneath damage sites. The integrity of the stomatal complex was severely compromised in damaged tissues. These histological changes suggest that damage by these mites will negatively impact the photosynthetic efficiency of susceptible plantation species.