Development of disease caused by Phytophthora cinnamomi in mature Xanthorrhoe australis

Over the past 30 years, heathland and open forest communities in south-eastern Australia dominated by Xanthorrhoea australis R.Br. have been severely affected by disease caused by Phytophthora cinnamomi Rands. The disease has caused a sharp decline in numbers of individuals within populations of X. australis; however, the etiology of the disease is unclear. The characteristics and disease symptoms induced by P. cinnamomi were analysed within nine mature X. australis plants that had been removed from the field. Seven plants showed typical disease symptoms that ranged from chlorotic leaves through to plant death. Plants showing disease symptoms had different numbers of infected roots, ranging from 0% in one dead plant, 40% infected roots in a plant showing yellowing of leaf tips and 67 and 86%, respectively, in two plants with severe chlorosis. There was variation within the roots, with some infected close to the stem while others were infected at more distal regions. Within stems of all plants, P. cinnamomi was difficult to isolate but was found in the desmium and stem apex and was associated with massive lesions within the central area of the stem. The symptoms of disease in X. australis are caused by a combination of damage to tissues of the roots and stem that may lead to a reduction in water and mineral transport throughout the plant.

Growth of Australian shortfin eel (Anguilla Australis) elvers given different dietary protein and lipid levels

The Australian shortfin eel, Anguilla australis is a potential candidate for intensive aquaculture. The present study was undertaken to evaluate the growth of elvers (5.4 g ± 0.1 initial weight) fed with diets of varying protein and lipid content, and to assess the potential of using soya-bean meal as a dietary ingredient. A 10 week experiment was conducted at 24 (±1.0) °C by rearing fish, in 60 L conical fibre glass tanks using a closed recirculation system. Diets having protein concentrations of 40 or 50% (by dry weight) were tested at three lipid levels (15, 20, 25%); diets being designated P₄₀L₁₅, P₄₀L₂₀, P₄₀L₂₅, P₅₀L₁₅, etc. All these diets contained 5% soya-bean meal. In addition P₅₀L₂₀ diets were formulated to contain 10 and 20% soya-bean meal in the diet (Diets S1 & S2). Shortfin eel grew best on the P₅₀L₁₅ diet, with an average specific growth rate of 2.26%. Food conservation ratio (FCR) and Protein efficiency ratio (PER) ranged from 1.21 (P₅₀L₁₅) to 2.12 (P₄₀L₂₅), and 0.92 (P₅₀L₂₅) to 1.65 (P₅₀L₁₅), respectively. Based on all criteria the best growth performance of shortfin eel was on the P₅₀L₁₅ diet, followed by P₄₀L₂₀ and P₄₀L₁₅. At both protein levels fish reared on diets with 25% lipid performed poorly. The performance of shortfin eel was not affected by the amount of soya-bean meal in the diet, up to a maximum of 20% dietary inclusion. No significant differences in muscle protein were evident in shortfin eel reared on different dietary treatments, nor was the lipid content of muscle related to dietary lipid level.

Weaning of Australian shortfin glass eels (Anguilla Australis): a comparison on the effectiveness of four types of fish roe

Eel culture is solely dependent on wild seed stock, caught in estuaries during the freshwater migratory phase as glass eels. The methods used for weaning glass eels are very variable, and range from the use of live zooplankton to fish roe to fines of commercial fish feeds. The present experiments were conducted on glass eels of the Australian shortfin eel, when the effectiveness of four types of readily available fish roe (European carp, mirror dory, orange roughy and warehou) were evaluated over a 42-day period, in the laboratory.

After 28 days the eels did not show an interest in orange roughy and mirror dory roe, and these two treatments were discontinued to avoid mortality. In all treatments there was a decrease in mean weight during this period, but the survival was over 99%. In the 28th to 42nd day period the mean weight and specific growth rate of glass eels reared on European carp and warehou roe increased, but the differences between these two treatments were not significant.

The physical features of the roe and the oocytes thereof, the proximate composition, amino acid and fatty acid composition indicated major differences amongst the roe types, particularly with regard to the amount of n−6 polyunsaturated fatty acids (PUFA) and the ratio of n−3 to n−6. European carp and warehou roe (and oocytes) had a significantly higher arachidonic acid (AA-20:4n−6; over 60% of PUFA) content and a considerably lower n−3 to n−6 ratio (n−3 to n−6 ratio being 1.32, 5.92, 3.77 and 2.67 for roe types, and 1.25, 4.83, 2.91 and 2.42 for oocytes, of European carp, mirror dory, orange roughy and warehou, respectively), than in the other two roe types. The fatty acid profiles of European carp and warehou roe were similar to that of metamorphosing glass eels.

M'Coy and the Australian Ichthyosaur Ichthyosaurus australis (M'Coy, 1867)

The first Australian ichthyosaur fossils were described by Frederick M'Coy in 1867 from a series of fossil specimens collected by James Sutherland in the Flinders River region, northern Queensland. An initial case of fossils collected was primarily used by M'Coy to provide the first incontrovertible proof of the existence of the Cretaceous System in Australia. Subsequent follow-up work was undertaken and further specimens were collected, including fossiI vertebrae that were named by M 'Coy, lchthyosaurus australis (M'Coy 1867). Despite describing the species as 'the most interesting fossil animal yet found in Australia' his descriptions were brief and limited and have been criticized by a number of later workers.

Lipid and fatty acid digestibility of three oil types in the Australian shortfin eel, Anguilla australis

The lipid and fatty acid digestibilities of three semi-purified, isonitrogenous (48.9–50.8% protein) and isocalorific (19.1–20.8 kJ g⁻¹) diets, in which the lipid source was either cod liver oil (CLO), linseed oil (LO) or sunflower oil (SFO), were estimated in the Australian shortfin eel (Anguilla australis) using chromic oxide as an external marker. Apparent percent protein and energy digestibilities of the diets were not significantly (P>0.05) affected by the lipid source, but the lipid digestibility was. The percent apparent lipid digestibility was lowest in the LO diet (90.2±0.6) and highest in the CLO diet (95.6±0.2).

Not all the fatty acids present in any one diet were recovered in the faecal samples. In diets with CLO, only three saturates (out of five), five monoenes and six (out of 11) PUFAs were detected in faecal samples. With all the diets, 20:0 and 22:0, and none of the n−6 HUFA were detected in the faecal samples. The digestibility of all the fatty acids, except 18:3n−3, was lowest in the diet with LO, and significantly so (P>0.05) from the other diets.

In shortfin eel, there was a trend for the digestibility of saturated fatty acids of diets with the animal oil as the lipid source to decrease with increasing chain length, and in diets with vegetable oil to increase initially and then decrease. A somewhat comparable trend was also evident in respect of monoenes.

When the digestibility of different categories of fatty acids is considered, the digestibility of saturates, monoenes, unsaturates, n−6, PUFA, HUFA and total fatty acid digestibilities of LO diet were the lowest, and differed significantly (P<0.05) from those of the CLO and SFO diets, except in the case of n−3 fatty acids.

Nitric oxide control of the dorsal aorta and the intestinal vein of the Australian short-finned eel Anguilla australis

This study investigated the mechanisms by which nitric oxide (NO) regulates the dorsal aorta and the intestinal vein of the Australian short-finned eel Anguilla australis. NADPH diaphorase histochemistry and immunohistochemistry using a mammalian endothelial nitric oxide synthase (NOS) antibody could not demonstrate NOS in the endothelium of either blood vessel; however, NOS could be readily demonstrated in the endothelium of the rat aorta that was used as a control. Both blood vessels contained NADPH diaphorase positive nerve fibres and nerve bundles, and immunohistochemistry using a neural NOS antibody showed a similar distribution of neural NOS immunoreactivity in the perivascular nerves. In vitro organ bath physiology showed that a NO/soluble guanylyl cyclase (GC) system is present in the dorsal aorta and the intestinal vein, since the soluble GC inhibitor oxadiazole quinoxalin^-1 (ODQ; 10^&ndash;5 mol l^&ndash;1) completely abolished the vasodilatory effect of the NO donor, sodium nitroprusside (SNP; 10^&ndash;4 mol l^&ndash;1). In addition, nicotine (3x10^&ndash;4 mol l^&ndash;1) mediated a vasodilation that was not affected by removal of the endothelium. The nicotine-mediated dilation was blocked by the NOS inhibitor, N&omega;-nitro-arginine (L-NNA; 10^&ndash;4 mol l^&ndash;1), and ODQ (10^&ndash;5 mol l^&ndash;1). More specifically, the neural NOS inhibitor, N&omega;-propyl-L-arginine (10^&ndash;5 mol l^&ndash;1), significantly decreased the dilation induced by nicotine (3x10^&ndash;4 mol l^&ndash;1). Furthermore, indomethacin (10^&ndash;5 mol l^&ndash;1) did not affect the nicotine-mediated dilation, suggesting that prostaglandins are not involved in the response. Finally, the calcium ionophore A23187 (3x10^&ndash;6 mol l^&ndash;1) caused an endothelium-dependent dilation that was abolished in the presence of indomethacin. We propose the absence of an endothelial NO system in eel vasculature and suggest that neurally derived NO contributes to the maintenance of vascular tone in this species. In addition, we suggest that prostaglandins may act as endothelially derived relaxing factors in A. australis.

Preliminary observations on the tank and pond culture of the glass eels of the Australian shortfin eel, Anguilla Australis Richardson

There is expanding interest in the culture of the Australian shortfin eel Anguilla australis Richardson; however, there is a lack of fundamental biology and husbandry information necessary to further develop an industry within Australia. The present study was undertaken to gain a preliminary understanding of basic husbandry requirements for rearing of juvenile A. australis (glass eels and elvers) in tanks and earthen ponds. Newly caught glass eels were successfully acclimated to culture conditions. During tank culture trials, specific growth rates (SGR) and survival rates ranged from −2.1 to 2.8% day⁻¹ and 52% to 100% respectively. Glass eels weaned onto a commercial eel diet exhibited a significantly greater SGR and survival rate than those weaned onto a commercial trout diet. Glass eels weaned onto an eel diet over a 15-day period grew slightly faster than eels weaned over a 5-day period, but survival rates were not significantly different for each treatment. SGRs (up to 2.8% day⁻¹) were significantly higher for glass eels fed at 9 and 12% day⁻¹ than at 6% day⁻¹. Stocking densities between 2.5 kg m⁻³ and 30 kg m⁻³ did not influence either SGR or survival rates. SGRs were significantly higher for glass eels cultured at 25 °C than at lower temperatures. During pond culture trials, SGRs and survival rates ranged from 1.36 to 1.65% day⁻¹ and 39% to 77% respectively. The SGR and survival rates of juvenile eels stocked into ponds receiving supplementary feeding with a commercial eel diet were not significantly different to those of eels stocked into ponds that did not receive supplementary feeding.

Immunolocalization of Na+/K+-ATPase, carbonic anhydrase II, and vacuolar H+-ATPase in the gills of freshwater adult lampreys, Geotria australis

As adults, anadromous lampreys migrate from seawater into freshwater rivers, where they require branchial ion (NaCl) absorption for osmoregulation. In teleosts and elasmobranchs, pharmological, immunohistochemical, and molecular data support roles for Na<sup>+/K⁺-ATPase (NPPase), carbonic anhydrase II (CAII), and vacuolar H⁺-ATPase (V-ATPase) in two different models of branchial ion absorption. To our knowledge, these transport-related proteins have not been studied in adult freshwater lampreys, and therefore it is not known if they are expressed, or have similar functions, in lampreys. The purpose of this study was to localize NPPase, CAII, and V-ATPase in the gills of adult freshwater lampreys and determine if any of these transport-related proteins are expressed in the same cells. Heterologous antibodies were used to localize the three proteins in gill tissue from pouched lamprey (Geotria australis). Immunoreactivity (IR) for all three proteins occurred between, and at the base of, lamellae in cells that match previous descriptions of mitochondrion-rich-cells (MRCs). NPPase-IR was always on the basolateral side of cells that did not stain for CAII or V-ATPase. In contrast, CAII-IR was always on the apical side of cells that also contained diffuse V-ATPase-IR. Therefore, we have identified two types of MRC in adult freshwater lamprey gills based on immunohistochemical staining for three transport proteins. A model of ion transport, based on our results, is proposed for adult freshwater lampreys. 

Potassium phosphonate alters the defence response of Xanthorrhoea australis following infection by Phytophthora cinnamomi

Potassium phosphonate (phosphite) is widely used in the management of Phytophthora diseases in agriculture, horticulture and natural environments. The Austral grass tree, Xanthorrhoea australis, a keystone species in the dry sclerophyll forests of southern Australia, is susceptible to Phytophthora cinnamomi, but is protected by applications of phosphite. We examined the effect of phosphite application on the infection of X. australis seedlings and cell suspension cultures by zoospores of P. cinnamomi. Phosphite induced more intense cellular responses to pathogen challenge and suppressed pathogen ingress in both seedlings and cell cultures. In untreated X. australis seedlings, hyphal growth was initially intercellular, became intracellular 24 h after inoculation, and by 48 h had progressed into the vascular tissue. In phosphite-treated seedlings, growth of P. cinnamomi remained intercellular and was limited to the cortex, even at 72 h after inoculation. The cell membrane retracted from the cell wall and phenolic compounds and electron dense substances were deposited around the wall of infected and neighbouring cells. Suspension cells were infected within 6 h of inoculation. Within 24 h of inoculation, untreated cells were fully colonised, had collapsed cytoplasm and died. The protoplast of phosphite-treated suspension cells collapsed within 12 h of inoculation, and phenolic material accumulated in adjacent, uninfected cells. No anatomical response to phosphite treatment was observed before infection of plant tissues, suggesting that the phosphite-associated host defence response is induced following pathogen challenge. Anatomical changes provide evidence that phosphite stimulates the host defence system to respond more effectively to pathogen invasion.

Weaning and growth of Anguilla australis glass eels and elvers

Anguilla australis glass eels proved to be resilient and present strong aquaculture potential. General husbandry techniques, anaesthesia and prophylactic treatments were established for glass eels between 0.1 g and 2.0 g and elvers between 2.0 g and 8.0 g, caught in rivers and estuaries along the South East Coast of Victoria. The protozoan parasites Ichthyobodo and Trichodina were found to be present on arrival to the hatchery developed during different rearing treatments, and were successfully eradicated. A. australis glass eels accepted artificial food, but it was recommended first be fed a preweaning diet of minced fish flesh. A weaning regime from minced fish flesh to commercially available eel grower mash, over 15 days was established. Growth rate proved to be highly variable, both between and within groups. The highest growth rate of 2.71%/day was found when the natural diet of minced fish and Artemia was fed. The maximum growth rate when reared on an artificial diet of 1.63%/day was observed at 25°C. Growth was affected by the presence or absence of a preweaning diet, weaning diet, weaning period, temperature, but not by size or density. Once weaned, glass eels were found to perform better on commercially available grower mash than on the minced fish flesh, which was used to aid in weaning them to artificial diets. Of the water quality parameters measured stocking density was found to affect pH, Total Ammonia Nitrogen, Total Phosphorus, and Dissolved Oxygen, through not to an extent which affected growth.

Aspects of the interaction between Xanthorrhoea australis and Phytophthora cinnamomi in south-western Victoria, Australia

Diseases in natural ecosystems are often assumed to be less severe than those observed in domestic cropping systems due to the extensive biodiversity exhibited in wild vegetation communities. In Australia, it is this natural biodiversity that is now under threat from Phytophthora cinnamomi. The soilborne Oomycete causes severe decline of native vegetation communities in south-western Victoria, Australia, disrupting the ecological balance of native forest and heathland communities. While the effect of disease caused by P. cinnamomi on native vegetation communities in Victoria has been extensively investigated, little work has focused on the Anglesea healthlands in south-western Victoria. Nothing is known about the population structure of P. cinnamomi at Anglesea. This project was divided into two main components to investigate fundamental issues affecting the management of P. cinnamomi in the Anglesea heathlands. The first component examined the phenotypic characteristics of P. cinnamomi isolates sampled from the population at Anglesea, and compared these with isolates from other regions in Victoria, and also from Western Australia. The second component of the project investigated the effect of the fungicide phosphonate on the host response following infection by P. cinnamomi. Following soil sampling in the Anglesea heathlands, a collection of P, cinnamomi isolates was established. Morphological and physiological traits of each isolate were examined. All isolates were found to be of the A2 mating type. Variation was demonstrated among isolates in the following characteristics: radial growth rate on various nutrient media, sporangial production, and sporangial dimensions. Oogonial dimensions did not differ significantly between isolates. Morphological and physiological variation was rarely dependant on isolate origin. To examine the genetic diversity among isolates and to determine whether phenotypic variation observed was genetically based, Random Amplified Polymorphic DNA (RAPD) analyses were conducted. No significant variation was observed among isolates based on an analysis of molecular variance (AMQVA). The results are discussed in relation to population biology, and the effect of genetic variation on population structure and population dynamics. X australis, an arborescent monocotyledon indigenous to Australia, is highly susceptible to infection by P. cinnamomi. It forms an important component of the heathland vegetation community, providing habitat for native flora and fauna, A cell suspension culture system was developed to investigate the effect of the fungicide phosphonate on the host-pathogen interaction between X. australis and P. cinnamomi. This allowed the interaction between the host and the pathogen to be examined at a cellular level. Subsequently, histological studies using X. australis seedlings were undertaken to support the cellular study. Observations in the cell culture system correlated well with those in the plant. The anatomical structure of X australis roots was examined to assist in the interpretation of results of histopathological studies. The infection of single cells and roots of X. australis, and the effect of phosphonate on the interaction are described. Phosphonate application prior to inoculation with P. cinnamomi reduced the infection of cells in culture and of cells in planta. In particular, phosphonate was found to stimulate the production of phenolic material in roots of X australis seedlings and in cells in suspension cultures. In phosphonate-treated roots of X australis seedlings, the deposition of electron dense material, possibly lignin or cellulose, was observed following infection with P. cinnamomi. It is proposed that this is a significant consequence of the stimulation of plant defence pathways by the fungicide. Results of the study are discussed in terms of the implications of the findings on management of the Anglesea heathlands in Victoria, taking into account variation in pathogen morphology, pathogenicity and genotype. The mode of action of phosphonate in the plant is discussed in relation to plant physiology and biochemistry.

Singing as a handicap : the effects of food availability and weather on song output in the Australian reed warbler Acrocephalus australis

Bird song is generally regarded as a sexually selected trait, and may represent a reliable handicap signal under at least certain conditions. Females may use the degree of male song production as a reliable cue to male condition or territory quality. We investigated the effect of supplementary feeding on song output in the migratory Australian reed warbler Acrocephalus australis. We experimentally increased the food availability on alternate days, and recorded several weather variables. We measured song rate and song length independently. Supplementary fed birds sang more on feeding days than on non-feeding days, while control birds did not show this effect. Song output was not significantly associated with any of the weather variables examined. Our results indicate that singing has the potential to serve as a reliable handicap signal to territorial food availability irrespective of the prevailing weather conditions. We discuss the role of energetic constraints and behavioural flexibility on the signaling function of song.

The functional significance of multiple nest-building in the Australian Reed Warbler Acrocephalus australis

The vast majority of bird species build a nest in which to breed. Some species build more than one nest, but the function of most multiple nest-building remains unclear. Here we describe the unusual nest-building behaviour of the Australian Reed Warbler Acrocephalus australis, and test experimentally the hypotheses that multiple nest-building is related to individual condition or territory quality, and plays a role in mate assessment. Australian Reed Warblers built two types of nest structures: 'type I' nests, which were used for eggs and nestlings, and 'type II' nests, which were structurally distinct from type I nests, did not support eggs, nestlings or adults and were not essential for successful breeding. The number of type II nests built in each territory varied. Type II nests were only built before breeding had commenced in a territory and females were not observed participating in their construction, supporting a role in female mate choice. Birds provided with supplementary food built significantly more type II nests than control birds. However, supplementary-fed birds did not have greater pairing success, and the addition of further type II nests to territories did not increase the pairing rate or type II nest construction in those territories. There was no relationship between the presence of type II nests and either reproductive success or likelihood of nest predation. We discuss the implications of these results in light of previous suggestions regarding the function of multiple nest-building in birds.