Propagation of Wollemi pine from tip cuttings and lower segment cuttings does not require rooting hormones

Domestication of the recently discovered and highly endangered Wollemi pine has relied almost entirely upon serial propagation of cuttings from a very small conservation collection. This study assessed the requirement for applied auxin to induce rooting in tip cuttings and lower segment cuttings of Wollemi pine. Both types of cuttings proved easy-to-root, with mean rooting of 71% for tip cuttings and 82% for lower segments. Auxin application (at 1.5, 3.0 or 8.0 g indole-3-butyric acid/L) did not accelerate root protrusion from propagation tubes or affect final rooting percentages.

Drosophila melanogaster mounts a unique immune response to the rhabdovirus Sigma virus

Rhabdoviruses are important pathogens of humans, livestock, and plants that are often vectored by insects. Rhabdovirus particles have a characteristic bullet shape with a lipid envelope and surface-exposed transmembrane glycoproteins. Sigma virus (SIGMAV) is a member of the Rhabdoviridae and is a naturally occurring disease agent of Drosophila melanogaster. The infection is maintained in Drosophila populations through vertical transmission via germ cells. We report here the nature of the Drosophila innate immune response to SIGMAV infection as revealed by quantitative reverse transcription-PCR analysis of differentially expressed genes identified by microarray analysis. We have also compared and contrasted the immune response of the host with respect to two nonenveloped viruses, Drosophila C virus (DCV) and Drosophila X virus (DXV). We determined that SIGMAV infection upregulates expression of the peptidoglycan receptor protein genes PGRP-SB1 and PGRP-SD and the antimicrobial peptide (AMP) genes Diptericin-A, Attacin-A, Attacin-B, Cecropin-A1, and Drosocin. SIGMAV infection did not induce PGRP-SA and the AMP genes Drosomycin-B, Metchnikowin, and Defensin that are upregulated in DCV and/or DXV infections. Expression levels of the Toll and Imd signaling cascade genes are not significantly altered by SIGMAV infection. These results highlight shared and unique aspects of the Drosophila immune response to the three viruses and may shed light on the nature of the interaction with the host and the evolution of these associations.

Apparent digestibility of some nutrient sources by juvenile mud crab, Scylla serrata (Forskal 1775)

As part of preliminary work aimed at the development of a formulated diet for the mud crab, Scylla serrata, an experiment was conducted with juvenile mud crabs (95.65±2.17 g) to determine apparent digestibility coefficients (ADC) for cellulose, fish meal, shrimp meal, blood meal, soybean meal, wheat flour and cod liver oil. Apparent digestibility coefficients for dry matter (ADCdm), energy (ADCenergy) and protein (ADC protein) were in the ranges 70.0-95.7%, 77.4-97.1% and 57.7-97.9% respectively. Soybean meal had the highest ADCdm and wheat flour had the lowest value (P<0.05), while the ADCdm for fish meal, blood meal and shrimp meal were not different (P?0.05). Similarly, soybean meal had the same ADCenergy as that of fish meal, but higher than those of cod liver oil, blood meal and shrimp meal (P<0.05). Moreover, the ADC protein for blood meal or shrimp meal were not significantly different from fish meal (P?0.05); nevertheless, they were lower than that of soybean meal and higher than that of wheat flour (P<0.05). Of significant interest was the ADCdm (78.0%) and ADCenergy (77.4%) for cellulose, which indicates that plant-based nutrient sources may well be a useful component of formulated diets for mud crabs.

An evaluation of the apxIVA based PCR-REA method for differentiation of Actinobacillus pleuropneumoniae

A restriction analysis of PCR (PCR-REA) amplified apxIVA gene has been suggested as an alternative method for serotyping of Actinobacillus pleuropneumoniae by Jaglic et al. [Jaglic, Z., Svastova, P., Rychlik, I., Nedbalcova, K., Kucerova, Z., Pavlik, I., Bartos, M., 2004. Differentiation of Actinobacillus pleuropneumoniae by PCR-REA based on sequence variability of the apxIVA gene and by ribotyping. Vet. Microbiol. 103, 63-69]. The current study investigated whether this alternative method could distinguish between the reference strains of serovars 13-15 and the value of the method when applied to 47 field isolates representing serovars 1-3, 5, 7-9, 12 and 15 as well as non-typable isolates. The reference strains of serovars 13 and 14 had the same sized product after the apxIVA PCR, while the product for serovar 15 was of different size compared to all the other serovar reference strains. The CfoI digest profiles of the reference serovars 13 and 14 strains were different from each other and from all other serovars. The HpaII digest profiles of these two serovars were very similar to each other, but both were distinctively different from the other serovar profiles. The CfoI digest profile of serovar 15 strain was very similar to the serovars 3 and 12 strains except for two faint extra bands for serovar 15. The HpaII digest profiles of serovars 12 and 15 reference strains were identical. The PCR-REA method correctly recognized the serovar of 21 of 43 field isolates. It was concluded that the method was a useful additional tool to support, but could not replace, conventional serotyping.

Comparison of sampling sites and detection methods for Haemophilus parasuis

Objective To improve the isolation rate and identification procedures for Haemophilus parasuis from pig tissues. Design Thirteen sampling sites and up to three methods were used to confirm the presence of H. parasuis in pigs after experimental challenge. Procedure Colostrum-deprived, naturally farrowed pigs were challenged intratracheally with H parasuis serovar 12 or 4. Samples taken during necropsy were either inoculated onto culture plates, processed directly for PCR or enriched prior to being processed for PCR. The recovery of H parasuis from different sampling sites and using different sampling methods was compared for each serovar. Results H parasuis was recovered from several sample sites for all serovar 12 challenged pigs, while the trachea was the only positive site for all pigs following serovar 4 challenge. The method of solid medium culture of swabs, and confirmation of the identity of cultured bacteria by PCR, resulted in 38% and 14% more positive results on a site basis for serovars 12 and 4, retrospectively, than direct PCR on the swabs. This difference was significant in the serovar 12 challenge. Conclusion Conventional culture proved to be more effective in detecting H parasuis than direct PCR or PCR on enrichment broths. For subacute (serovar 4) infections, the most successful sites for culture or direct PCR were pleural fluid, peritoneal fibrin and fluid, lung and pericardial fluid. For acute (serovar 12) infections, the best sites were lung, heart blood, affected joints and brain. The methodologies and key sampling sites identified in this study will enable improved isolation of H parasuis and aid the diagnosis of Glässer's disease.

The effects of salivary gland extracts from Boophilus microplus ticks on mitogen-stimulated bovine lymphocytes

The effect of salivary gland extract (SGE) from the tick Boophilus microplus was examined in mitogen-stimulated lymphocytes in vitro. SGE was added to lymphocytes of seven cattle together with the mitogens concanavalin A (ConA), phytohaemagglutinin (PHA) and pokeweed mitogen (PWM). Semi-purified B cells from another seven cattle were stimulated with the mitogen lipopolysaccharide (LPS). PHA and ConA stimulated proliferation of lymphocytes to the same extent, but the inhibition due to SGE of Boophilus microplus on the proliferative response stimulated by PHA (39.0% ± 9.3%) was less than the inhibition of proliferative response stimulated by ConA (75.4% ± 6.9%). In contrast, SGE of B. microplus stimulated the proliferation of B cells in the presence of LPS in a dose-dependent manner. Enhanced stimulation of B cells by SGE at >4 μg in culture was greater than twice that observed when B cells were stimulated by LPS alone. SGE does not have a direct suppressive effect on bovine B cell proliferation; however, in vivo the effectiveness of B cell responses might be influenced by other immune factors, such as cytokine profiles.

Long-term impacts of harvest residue management on nutrition, growth and productivity of an exotic pine plantation of sub-tropical Australia

Residue retention is an important issue in evaluating the sustainability of production forestry. However, its long-term impacts have not been studied extensively, especially in sub-tropical environments. This study investigated the long-term impact of harvest residue retention on tree nutrition, growth and productivity of a F1 hybrid (Pinus elliottii var. elliottii × Pinus caribaea var. hondurensis) exotic pine plantation in sub-tropical Australia, under three harvest residue management regimes: (1) residue removal, RR0; (2) single residue retention, RR1; and (3) double residue retention, RR2. The experiment, established in 1996, is a randomised complete block design with 4 replicates. Tree growth measurements in this study were carried out at ages 2, 4, 6, 8 and 10 years, while foliar nutrient analyses were carried out at ages 2, 4, 6 and 10 years. Litter production and litter nitrogen (N) and phosphorus (P) measurements were carried out quarterly over a 15-month period between ages 9 and 10 years. Results showed that total tree growth was still greater in residue-retained treatments compared to the RR0 treatment. However, mean annual increments of diameter at breast height (MAID) and basal area (MAIB) declined significantly after age 4 years to about 68-78% at age 10 years. Declining foliar N and P concentrations accounted for 62% (p < 0.05) of the variation of growth rates after age 4 years, and foliar N and P concentrations were either marginal or below critical concentrations. In addition, litter production, and litter N and P contents were not significantly different among the treatments. This study suggests that the impact of residue retention on tree nutrition and growth rates might be limited over a longer period, and that the integration of alternative forest management practices is necessary to sustain the benefits of harvest residues until the end of the rotation.

Light environment and tree development of young Acacia melanoxylon in mixed-species regrowth forest, Tasmania, Australia

Blackwood (Acacia melanoxylon R. Br.) is a valuable leguminous cabinetwood species which is commonly found as a canopy or subcanopy tree in a broad range of mixed-species moist forests on tablelands and coastal escarpments in eastern Australia. This paper reports on the competitive light environment of a commercially valuable multi-species regrowth forest in NW Tasmania, in order to define some of the functional interactions and competitive dynamics of these stands. Comparative observations were made of the internal forest light environment in response to small-gap silvicultural treatments, in a young regenerative mix of three codominant tree species. Light measurements were made during periods of maximum external irradiance of the regrowth Eucalyptus obliqua/A. melanoxylon forest canopy at age 10.5 years. This was at a time of vigourous stand development, 4.5 years following the application of three experimental silvicultural treatments whose effects were observed in comparison with an untreated canopy sample designed as a control. Minimal irradiance was observed within and beneath the dense subcanopy of the native nurse species (Pomaderris apetala) which closely surrounds young blackwood regeneration. Unlike current plantation nurse systems, the dense foliage of the native broadleaved Pomaderris all but eliminated direct side-light and low-angle illumination of the young blackwood, from the beginning of tree establishment. The results demonstrated that retention of these densely stocked native codominants effectively suppressed both size and frequency of blackwood branches on the lower bole, through effective and persistent interception of sunlight. Vigorous young blackwood crowns later overtopped the codominant nurse species, achieving a predictable height of branch-free bole. This competitive outcome offers a valuable tool for management of blackwood crown dynamics, stem form and branch habit through manipulation of light environment in young native regrowth systems. Results demonstrate that effective self-pruning in the lower bole of blackwood is achieved through a marked reduction in direct and diffuse sunlight incident on the lower crown, notably to less than 10-15% of full sunlight intensity during conditions of maximum insolation. The results also contain insights for the improved design of mixed-species plantation nurse systems using these or functionally similar species' combinations. Based on evidence presented here for native regrowth forest, plantation nurse systems for blackwood will need to achieve 85-90% interception of external side-light during early years of tree development if self-pruning is to emulate the results achieved in the native nurse system.

Ceratocystis atrox sp. nov. associated with Phoracantha acanthocera infestations on Eucalyptus grandis in Australia

Ceratocystis spp. include important pathogens of trees as well as apparently saprophytic species. Four species have been recorded on Eucalyptus grandis in Australia, of which only one, C. pirilliformis Barnes and M.J. Wingf., is known to be pathogenic. A recent survey of pests and diseases of Eucalyptus trees in northern Queensland revealed a species of Ceratocystis associated with the tunnels made by the aggressive wood-boring insect Phoracantha acanthocera (Macleay) (Cerambicydae: Coleoptera). The aim of the present study was to identify the fungus based on morphological characteristics and comparisons of DNA sequence data for three gene regions. The fungus peripherally resembles C. fimbriata Ell. and Halst. but differs from this species most obviously by having much darker mycelium, longer ascomatal necks, segmented hyphae and an absence of aleuroconidia. Comparisons of combined sequence data confirmed that the Ceratocystis sp. from P. acanthocera represents an undescribed taxon, which is provided with the name Ceratocystis atrox sp. nov. C. atrox appears to have a close relationship with P. acanthocera, although its role in the biology of the insect is unknown and its pathogenicity has not been considered.

Salmonella and on-farm risk factors in healthy slaughter-age cattle and sheep in eastern Australia

To examine healthy slaughter-age cattle and sheep on-farm for the excretion of Salmonella serovars in faeces and to identify possible risk factors using a questionnaire. The study involved 215 herds and flocks in the four eastern states of Australia, 56 with prior history of salmonellosis. Production systems examined included pasture beef cattle, feedlot beef cattle, dairy cattle, prime lambs and mutton sheep and animals were all at slaughter age. From each herd or flock, 25 animals were sampled and the samples pooled for Salmonella culture. All Salmonella isolated were serotyped and any Salmonella Typhimurium isolates were phage typed. Questionnaires on each production system, prepared in Epi Info 6.04, were designed to identify risk factors associated with Salmonella spp excretion, with separate questionnaires designed for each production system. Salmonellae were identified in all production systems and were more commonly isolated from dairies and beef feedlots than other systems. Statistical analysis revealed that dairy cattle were significantly more likely to shed Salmonella in faeces than pasture beef cattle, mutton sheep and prime lambs (P < 0.05). A wide diversity of Salmonella serovars, all of which have been isolated from humans in Australia, was identified in both cattle and sheep. Analysis of the questionnaires showed access to new arrivals was a significant risk factor for Salmonella excretion on dairy properties. For beef feedlots, the presence of large numbers of flies in the feedlot pens or around stored manure were significant risk factors for Salmonella excretion. Dairy cattle pose the highest risk of all the slaughter-age animals tested. Some of the identified risk factors can be overcome by improved management practices, especially in relation to hygiene.

Building better information products by assessing the information needs of growers of subtropical strawberries in Queensland

Timely access to effective technical information is a key ingredient of profitable strawberry production. Through the Better Berries Program, a joint RD&E initiative of government and industry, a number of information products and services have been provided in recent years to Australia's subtropical strawberry industry, centred in southern Queensland. However, there is a lack of knowledge of how well these are meeting the information needs of growers, both in content and delivery. To better understand grower information use and needs, a stratified sample of 25 growers was interviewed on-farm during the 2004 season. Growers were asked about how they currently accessed information, what they thought of a range of information products and ideas on show, and what advice they would provide for information development in the future. Results indicated that information sought by growers and the style in which it is best presented, varied considerably with grower experience, but little with farm size. New growers had a wide range of needs while the needs of experienced growers were focused mainly on problem identification and new production development. Interestingly, the overwhelming majority across all sectors still preferred paper-based information products despite their extensive use of computers for business purposes. The findings were used to develop a strategy for an improved range of technical information products and services that are more accessible, easier to use, more timely, and more relevant to the needs of growers.

Concordant molecular and phenotypic data delineate new taxonomy and conservation priorities for the endangered mountain yellow-legged frog

The mountain yellow-legged frog Rana muscosa sensu lato, once abundant in the Sierra Nevada of California and Nevada, and the disjunct Transverse Ranges of southern California, has declined precipitously throughout its range, even though most of its habitat is protected. The species is now extinct in Nevada and reduced to tiny remnants in southern California, where as a distinct population segment, it is classified as Endangered. Introduced predators (trout), air pollution and an infectious disease (chytridiomycosis) threaten remaining populations. A Bayesian analysis of 1901 base pairs of mitochondrial DNA confirms the presence of two deeply divergent clades that come into near contact in the Sierra Nevada. Morphological studies of museum specimens and analysis of acoustic data show that the two major mtDNA clades are readily differentiated phenotypically. Accordingly, we recognize two species, Rana sierrae, in the northern and central Sierra Nevada, and R. muscosa, in the southern Sierra Nevada and southern California. Existing data indicate no range overlap. These results have important implications for the conservation of these two species as they illuminate a profound mismatch between the current delineation of the distinct population segments (southern California vs. Sierra Nevada) and actual species boundaries. For example, our study finds that remnant populations of R. muscosa exist in both the southern Sierra Nevada and the mountains of southern California, which may broaden options for management. In addition, despite the fact that only the southern California populations are listed as Endangered, surveys conducted since 1995 at 225 historic (1899-1994) localities from museum collections show that 93.3% (n=146) of R. sierrae populations and 95.2% (n=79) of R. muscosa populations are extinct. Evidence presented here underscores the need for revision of protected population status to include both species throughout their ranges.

Influence of food supplementation on the fitness of two biological control agents: a predatory nabid bug and a bollworm pupal parasitoid

Many arthropod predators and parasitoids exhibit either stage-specific or lifetime omnivory, in that they include extra-floral nectar, floral nectar, honeydew or pollen in their immature and/or adult diet. Access to these plant-derived foods can enhance pest suppression by increasing both the individual fitness and local density of natural enemies. Commercial products such as Amino-Feed®, Envirofeast®, and Pred-Feed® can be applied to crops to act as artificial-plant-derived foods. In laboratory and glasshouse experiments we examined the influence of carbohydrate and protein rich Amino-Feed UV® or Amino-Feed, respectively, on the fitness of a predatory nabid bug Nabis kinbergii Reuter (Hemiptera: Nabidae) and bollworm pupal parasitoid Ichneumon promissorius (Erichson) (Hymenoptera: Ichneumonidae). Under the chosen conditions, the provision of either wet or dry residues of Amino-Feed UV had no discernable effect on immediate or longer-term survival and immature development times of N. kinbergii. In contrast, the provision of honey, Amino-Feed plus extrafloral nectar, and extrafloral nectar alone had a marked effect on the longevity of I. promissorius, indicating that they were limited by at least carbohydrates as an energy source, but probably not protein. Compared with a water only diet, the provision of Amino-Feed plus extrafloral nectar increased the longevity of males and females of I. promissorius by 3.0- and 2.4-fold, respectively. Not only did female parasitoids live longer when provided food, but the total number of eggs laid and timing of deposition was affected by diet under the chosen conditions. Notably, females in the water and honey treatments deposited greater numbers of eggs earlier in the trial, but this trend was unable to be sustained over their lifetime. Egg numbers in these treatments subsequently fell below the levels achieved by females in the Amino-Feed plus extrafloral nectar and cotton extrafloral nectar only treatments. Furthermore, there were times when the inclusion of the Amino-Feed was beneficial compared with cotton extrafloral nectar only. Artificial food supplements and plant-derived foods are worthy of further investigation because they have potential to improve the ecosystem service of biological pest control in targeted agroecosystems by providing natural enemies with an alternative source of nutrition, particularly during periods of prey/host scarcity.

Temporal variation in arthropod sampling effectiveness: The case for using the beat sheet method in cotton

Predatory insects and spiders are key elements of integrated pest management (IPM) programmes in agricultural crops such as cotton. Management decisions in IPM programmes should to be based on a reliable and efficient method for counting both predators and pests. Knowledge of the temporal constraints that influence sampling is required because arthropod abundance estimates are likely to vary over a growing season and within a day. Few studies have adequately quantified this effect using the beat sheet, a potentially important sampling method. We compared the commonly used methods of suction and visual sampling to the beat sheet, with reference to an absolute cage clamp method for determining the abundance of various arthropod taxa over 5 weeks. There were significantly more entomophagous arthropods recorded using the beat sheet and cage clamp methods than by using suction or visual sampling, and these differences were more pronounced as the plants grew. In a second trial, relative estimates of entomophagous and phytophagous arthropod abundance were made using beat sheet samples collected over a day. Beat sheet estimates of the abundance of only eight of the 43 taxa examined were found to vary significantly over a day. Beat sheet sampling is recommended in further studies of arthropod abundance in cotton, but researchers and pest management advisors should bear in mind the time of season and time of day effects.

Pest management in Queensland strawberries: Reality bites, and growers' perspectives change!

Prior to the 1980s, arthropod pest control in Queensland strawberries was based entirely on calendar sprays of insecticides (mainly endosulfan, triclorfon, dimethoate and carbaryl) and a miticide (dicofol). These chemicals were applied frequently and spider mite outbreaks occurred every season. The concept of integrated pest management (IPM) had not been introduced to growers, and the suggestion that an alternative to the standard chemical pest control recipe might be available, was ignored. Circumstances changed when the predatory mite, Phytoseiulus persimilis Athios-Henriot, became available commercially in Australia, providing the opportunity to manage spider mites, the major pests of strawberries, with an effective biological agent. Trials conducted on commercial farms in the early 1980s indicated that a revolution in strawberry pest management was at hand, but the industry generally remained sceptical and afraid to adopt the new strategy. Lessons are learnt from disasters and the consequent monetary loss that ensues, and in 1993, such an event relating to ineffective spider mite control, spawned the revolution we had to have. Farm-oriented research and evolving grower perspectives have resulted in the acceptance of biological control of spider mites using Phytoseiulus persimilis and the 'pest in first' technique, and it now forms the basis of an IPM system that is used on more than 80% of the Queensland strawberry crop.