Emerging henipaviruses and flying foxes - Conservation and management perspectives

Wildlife populations are affected by a series of emerging diseases, some of which pose a significant threat to their conservation. They can also be reservoirs of pathogens that threaten domestic animal and human health. In this paper, we review the ecology of two viruses that have caused significant disease in domestic animals and humans and are carried by wild fruit bats in Asia and Australia. The first, Hendra virus, has caused disease in horses and/or humans in Australia every five years since it first emerged in 1994. Nipah virus has caused a major outbreak of disease in pigs and humans in Malaysia in the late 1990s and has also caused human mortalities in Bangladesh annually since 2001. Increased knowledge of fruit bat population dynamics and disease ecology will help improve our understanding of processes driving the emergence of diseases from bats. For this, a transdisciplinary approach is required to develop appropriate host management strategies that both maximise the conservation of bat populations as well as minimise the risk of disease outbreaks in domestic animals and humans.

Recent advances in the molecular biology of Leifsonia xyli subsp. xyli, causal organism of ratoon stunting disease

Twelve years ago our understanding of ratoon stunting disease (RSD) was confined almost exclusively to diagnosis of the disease and control via farm hygiene, with little understanding of the biology of the interaction between the causal agent (Leifsonia xyli subsp. xyli) and the host plant sugarcane (Saccharum spp. hybrids). Since then, research has focused on developing the molecular tools to dissect L. xyli subsp. xyli, so that better control strategies can be developed to prevent losses from RSD. Within this review, we give a brief overview of the progression in research on L. xyli subsp. xyli and highlight future challenges. After a brief historical background on RSD, we discuss the development of molecular tools such as transformation and transposon mutagenesis and discuss the apparent lack of genetic diversity within the L. xyli subsp. xyli world population. We go on to discuss the sequencing of the genome of L. xyli subsp. xyli, describe the key findings and suggest some future research based on known deficiencies that will capitalise on this tremendous knowledge base to which we now have access.

Developing improved supply chains for temperate fruits in transitional Asian economies of Thailand and Vietnam.

Asia's increasing demand for both tropical and temperate fruit is projected to grow significantly. Compared with most developed countries, the production of temperate fruits (peach, nectarine, plum and apple) has expanded rapidly in China over the past 20 years. In contrast, current production of plums and peaches in neighbouring countries (Thailand and Vietnam) is very low but their fruit enters the market earlier. Thailand and Vietnam have enormous potential to satisfy a market window in the northern hemisphere period from March to May inclusive when there is little or no stone fruit on the Asian market. In Vietnam, fruit is harvested in an immature state to avoid disease and fruit fly problems and consequently lacks size and flavour. Approximately 30-40% of locally produced fruit in Vietnam does not reach market due to disease and poor handling during picking and transport. In Thailand, much of the infrastructure needed to transport, store, process and market temperate fruits successfully are now in place. However, there are currently no cool chain management or quality assurance systems to ensure a fresh product reaches the consumer with minimal deterioration. In Vietnam, growing stone fruit under the traditional system with little or minimal inputs, the farmer may receive between AUD3,000-5,000 per ha. In comparison, under higher input systems incorporating fertiliser, irrigation and pest and disease management, net returns can be increased seven-fold. Strengths and weaknesses of the current supply chains in these two countries are discussed.

The genetic diversity of ampeloviruses in Australian pineapples and their association with mealybug wilt disease

Pineapple mealybug wilt-associated virus 1 (PMWaV-1), 2 (PMWaV-2) and -3 (PMWaV-3) have been detected in Australian commercial pineapple crops, along with a previously undescribed ampelovirus, for which the name Pineapple mealybug wilt-associated virus 5 (PMWaV-5) is proposed. Partial sequences extending from open reading frame 1b through to the heat shock protein homologue were obtained for PMWaV-1, -3 and -5. Phylogenetic analyses of selected regions of these sequences indicated that PMWaV-5 is a distinct species and most closely related to PMWaV-1. The amino acid sequence variation observed in the RNA-dependent RNA polymerase region of PMWaV-1 isolates was 95.8–98.4% and of PMWaV-3 isolates was 92.2–99.5%. In surveys of mealybug wilt disease (MWD) affected crops, none of the four viruses was clearly associated with the disease at all survey sites. A statistically significant association (P < 0.001) between the presence of PMWaV-2 and symptoms was observed at one survey site (site 3), but the virus was at a low incidence at the remaining three survey sites. By contrast, although PMWaV-1 and -3 were equally distributed between symptomless and MWD-affected plants at site 3, there was a statistically significant (P < 0.001) association between each of these two viruses and MWD at sites 1 and 4. At site 2, there was a statistically significant (P < 0.001) association only between PMWaV-3 and MWD. PMWaV-1 was the most commonly found of the four viruses and conversely PMWaV-5 was only occasionally found. Australian isolates of PMWaV-1, -2 and -3 were transmitted by the mealybug species Dysmicoccus brevipes.

Experimental Studies on Drop Formation at the Tip of Melting Rods

Drop formation at the conical tips of melting rods has been experimentally studied using the transparent wax-alcohol/acetonitrile system. The effects of cone angle, rod diameter, immersion depth, and bath temperature on the detached drop mass have been studied over a wide range, besides recording useful qualitative information based on visual observation. The experimental results suggest that the phenomenon of drop formation at the tip of melting rods has a close parallel with the drop formation at conical tips, at least on a qualitative basis. However, the results could not be quantified owing to difficulties in characterizing the physical properties of the system, despite efforts to minimize them.

An absolute method for the determination of surface tension of liquids using pendent drop profiles

Drop formation at conical tips which is of relevance to metallurgists is investigated based on the principle of minimization of free energy using the variational approach. The dimensionless governing equations for drop profiles are computer solved using the fourth order Runge-Kutta method. For different cone angles, the theoretical plots of XT and ZT vs their ratio, are statistically analyzed, where XT and ZT are the dimensionless x and z coordinates of the drop profile at a plane at the conical tip, perpendicular to the axis of symmetry. Based on the mathematical description of these curves, an absolute method has been proposed for the determination of surface tension of liquids, which is shown to be preferable in comparison with the earlier pendent-drop profile methods.

Carotenoid content and flesh color of selected banana cultivars growing in Australia.

Background: The problems of vitamin A deficiency and chronic diseases have emerged in recent years in some countries in the Micronesian region. These problems are associated with the dietary shift towards imported processed foods and lifestyle changes. Research in the Federated States of Micronesia indicates that yellow- and orange-fleshed banana cultivars contain significant levels of provitamin A carotenoids. Objective: To identify further banana cultivars that may be promoted to alleviate vitamin A deficiency among children and women and chronic disease problems among adults. Methods: Ripe fruit of banana cultivars growing in Australia (sourced mostly from a field research collection) were assessed for carotenoid content and flesh color. Ten cultivars with yellow or yellow/orange flesh color (including common cultivars of Southeast Asia and the Pacific Islands) were selected and compared with two cream-fleshed cultivars, including Williams, of the Cavendish group, the most commonly marketed banana worldwide. Carotenoid content was analyzed by high-performance liquid chromatography (HPLC). Flesh color was analyzed by HunterLab colorimetry. Results: The yellow/orange-fleshed Asupina (a Fe'i banana) contained the highest level (1,412 μg/100 g) of trans β-carotene, the most important provitamin A carotenoid, a level more than 20 times higher than that of Williams. All 10 yellow or yellow/orange-fleshed cultivars (Asupina, Kirkirnan, Pisang Raja, Horn Plantain, Pacific Plantain, Kluai Khai Bonng, Wain, Red Dacca, Lakatan, and Sucrier) had significant carotenoid levels, potentially meeting half or all of the estimated vitamin A requirements for a nonpregnant, nonlactating adult woman within normal consumption patterns. All were acceptable for taste and other attributes. The cream-fleshed cultivars had minimal carotenoid levels. There was a positive significant correlation between carotenoid content and deeper yellow/orange coloration indicators. Conclusions: These yellow- or yellow/orange-fleshed carotenoid-rich banana cultivars should be considered for promotion in order to alleviate vitamin A deficiency and chronic disease in susceptible target communities and to provide variety and enjoyment as exotic fruits in both developing and industrialized countries.

Genetic analysis of foot-and-mouth disease virus serotype A of Indian origin and detection of positive selection and recombination in leader protease- and capsid-coding regions

The leader protease (L-pro) and capsid-coding sequences (P1) constitute approximately 3 kb of the foot-and-mouth disease virus (FMDV). We studied the phylogenetic relationship of 46 FMDV serotype A isolates of Indian origin collected during the period 1968-2005 and also eight vaccine strains using the neighbour-joining tree and Bayesian tree methods. The viruses were categorized under three major groups - Asian, Euro-South American and European. The Indian isolates formed a distinct genetic group among the Asian isolates. The Indian isolates were further classified into different genetic subgroups (<5% divergence). Post-1995 isolates were divided into two subgroups while a few isolates which originated in the year 2005 from Andhra Pradesh formed a separate group. These isolates were closely related to the isolates of the 1970s. The FMDV isolates seem to undergo reverse mutation or onvergent evolution wherein sequences identical to the ancestors are present in the isolates in circulation. The eight vaccine strains included in the study were not related to each other and belonged to different genetic groups. Recombination was detected in the L-pro region in one isolate (A IND 20/82) and in the VP1 coding 1D region in another isolate (A RAJ 21/96). Positive selection was identified at aa positions 23 in the L-pro (P<0.05; 0.046*) and at aa 171 in the capsid protein VP1 (P<0.01; 0.003**).

Cultivar affects browning susceptibility of freshly cut star fruit slices

Consumption of freshly-cut horticultural products has increased in the last few years. The principal restraint to using freshly-cut carambola is its susceptibility to tissue-browning, due to polyphenol oxidase-mediated oxidation of phenolic compounds present in the tissue. The current study investigated the susceptibility to browning of star fruit slices (Averrhoa carambola L.) of seven genotypes (Hart, Golden Star, Taen-ma, Nota-10, Malasia, Arkin, and Fwang Tung). Cultivar susceptibility to browning as measured by luminosity (L*) varied significantly among genotypes. Without catechol 0.05 M, little changes occurred on cut surface of any cultivars during 6 hour at 25 degrees C, 67% RH. Addition of catechol led to rapid browning, which was more intense in cvs. Taen-ma, Fwang Tung, and Golden Star, with reduction in L* value of 28.60%, 27.68%, and 23.29%, respectively. Browning was more intense in the center of the slices, particularly when treated with catechol, indicating highest polyphenol oxidase (PPO) concentration. Epidermal browning, even in absence of catechol, is a limitation to visual acceptability and indicates a necessity for its control during carambola processing. Care must be given to appropriate selection of cultivars for fresh-cut processing, since cultivar varied in browning susceptibility in the presence of catechol.

Effects of rootstock and nitrogen fertiliser on postharvest anthracnose development in Hass avocado

These rootstock and nitrogen fertiliser studies confirmed that rootstock race can significantly affect the development of postharvest disease and mineral nutrient accumulation in Hass avocado fruit. When Hass (Guatemalan race) was grafted to seedling Velvick (West Indian race) rootstock, the severity and incidence of anthracnose in fruit were significantly reduced by up to 64 and 37%, respectively, compared with seedling Duke 6 (Mexican race) rootstock. Stem-end rot was also influenced by rootstock in some seasons, and significant reductions (up to 87%) in the severity and incidence of stem-end rot were recorded in Hass fruit from Velvick compared with Duke 6 rootstock trees. These improvements in postharvest diseases were associated with significantly lower concentrations of nitrogen and potassium, higher concentrations of calcium and magnesium, lower ratios of nitrogen:calcium and higher ratios of calcium + magnesium:potassium in Hass leaves and fruit from Velvick compared with Duke 6 rootstock trees. Altering the rate of nitrogen fertiliser had minimal impact on postharvest disease development. However, in one season, reducing the rate of nitrogen fertiliser to nil significantly reduced the concentration of nitrogen in the fruit skin, decreased the nitrogen:calcium ratio and significantly reduced the severity and incidence of anthracnose in Hass fruit from both Velvick and Duke 6 rootstock trees. The form of nitrogen fertiliser (ammonium compared with nitrate) applied to the trees did not significantly affect the postharvest disease susceptibility of Hass avocado fruit on either Velvick or Duke 6 rootstock. The Guatemalan race rootstocks, Anderson 8 and Anderson 10, were also found to be superior to the Mexican race rootstock, Parida 1, for reducing anthracnose severity. This again, was associated with a better balance of mineral nutrients (significantly lower nitrogen:calcium and higher calcium + magnesium:potassium ratios) in the fruit. This rootstock effect, however, was only observed in the first season of a 3-year experiment, possibly because of a better balance between vegetative growth and fruit production in Parida 1 in the latter two seasons. Significant positive correlations between anthracnose severity and fruit skin nitrogen:calcium ratios were evident across all experiments.

Pimelea trichostachya poisoning (St George disease) in horses

A dense population of Pimelea trichostachya plants (Family Thymelaeaceae) in pasture poisoned a horse herd in southern inland Queensland in October-November 2005. Plant density was 2 to 45 g wet weight/m2 (mean 16 g/m2) from 5 to 69 plants/m2 (mean 38 plants/m2) representing 3 to 20% (mean 9%) of the volume of pasture on offer. Ten of 35 mares, fillies and geldings were affected. Clinical signs were loss of body weight, profound lethargy, serous nasal discharge, severe watery diarrhoea and subcutaneous oedema of the intermandibular space, chest and ventral midline. Pathological findings were anaemia, leucocytopenia, hypoproteinaemia, dilatation of the right ventricle of the heart, dilated hepatic portal veins and periportal hepatic sinusoids (peliosis hepatis), alimentary mucosal hyperaemia and oedema of mesenteric lymph nodes. Cattle grazing the same pasture were affected by Pimelea poisoning simultaneously. Removal of the horses to Pimelea-free pasture initiated recovery. The one other incident of this syndrome, previously only recognised in cattle in Australia, occurred in horses, in South Australia in 2002, with access to a dense Pimelea simplex population.

The mortality of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) neonate larvae in relation to drop-off and soil surface temperature: The dangers of bungy jumping

The first larval instar has been identified as a critical stage for population mortality in Lepidoptera, yet due to the body size of these larvae, the factors that contribute to mortality under field conditions are still not clear. Dispersal behaviour has been suggested as a significant, but ignored factor contributing to mortality in first-instar lepidopteran larvae. The impact that leaving the host plant has on the mortality rate of Helicoverpa armigera neonates was examined in field crops and laboratory trials. In this study the following are examined: (1) the effects of soil surface temperature, and the level of shade within the crop, on the mortality of neonates on the soil after dropping off from the host plant; (2) the percentage of neonates that dropped off from a host plant and landed on the soil; and (3) the effects of exposure to different soil surface temperatures on the development and mortality of neonates. The findings of this study showed that: (1) on the soil, surface temperatures above 43°C were lethal for neonates, and exposure to these temperatures contributed greatly to the overall mortality rate observed; however, the fate of neonates on the soil varied significantly depending on canopy closure within the crop; (2) at least 15% of neonates dropped off from the host plant and landed on the soil, meaning that the proportion of neonates exposed to these condition is not trivial; and (3) 30 min exposure to soil surface temperatures approaching the lethal level (>43°C) has no significant negative effects on the development and mortality of larvae through to the second instar. Overall leaving the plant through drop-off contributes to first-instar mortality in crops with open canopies; however, survival of neonates that have lost contact with a host plant is possible, and becomes more likely later in the crop growing season.

Characterization of disease resistance gene candidates of the nucleotide binding site (NBS) type from banana and correlation of a transcriptional polymorphism with resistance to Fusarium oxysporum f.sp. cubense race 4

Most plant disease resistance (R) genes encode proteins with a nucleotide binding site and leucine-rich repeat structure (NBS-LRR). In this study, degenerate primers were used to amplify genomic NBS-type sequences from wild banana (Musa acuminata ssp. malaccensis) plants resistant to the fungal pathogen Fusarium oxysporum formae specialis (f. sp.) cubense (FOC) race 4. Five different classes of NBS-type sequences were identified and designated as resistance gene candidates (RGCs). The deduced amino acid sequences of the RGCs revealed the presence of motifs characteristic of the majority of known plant NBS-LRR resistance genes. Structural and phylogenetic analyses grouped the banana RGCs within the non-TIR (homology to Toll/interleukin-1 receptors) subclass of NBS sequences. Southern hybridization showed that each banana RGC is present in low copy number. The expression of the RGCs was assessed by RT-PCR in leaf and root tissues of plants resistant or susceptible to FOC race 4. RGC1, 3 and 5 showed a constitutive expression profile in both resistant and susceptible plants whereas no expression was detected for RGC4. Interestingly, RGC2 expression was found to be associated only to FOC race 4 resistant lines. This finding could assist in the identification of a FOC race 4 resistance gene.

Tan spot: A new disease of pyrethrum caused by Microsphaeropsis tanaceti sp. nov

The isolation frequency of Microsphaeropsis sp. in spring in association with necrotic lesions on leaves in Tasmanian pyrethrum (Tanacetum cinerariifolium) fields has increased substantially since first identification in 2001. Examination of morphological features and sequencing of the internal transcribed spacer region (ITS) resulted in the identification of a new species, herein described as Microsphaeropsis tanaceti sp. nov. The pathogenicity of three M. tanaceti isolates to two pyrethrum cultivars was confirmed by inoculating glasshouse-grown plants in three experiments. No significant differences in the susceptibility of the two cultivars to infection by M. tanaceti were found. Symptoms were tan-coloured spots which coalesced around the margins of the leaves. Therefore, the name 'tan spot' is proposed for this new disease of pyrethrum. The sensitivity of seven M. tanaceti isolates to difenoconazole and azoxystrobin, commonly used fungicides for the management of foliar diseases in spring, was assessed under in vitro conditions. Sensitivity testing for difenoconazole was conducted using a mycelial growth assay on potato dextrose agar, whilst testing for sensitivity to azoxystrobin used a conidial germination assay on water agar. Microsphaeropsis tanaceti was found to be more sensitive to azoxystrobin than difenoconazole, with complete inhibition of conidial germination at concentrations above 0.625 µg a.i. mL-1. By comparison, concentrations of 50 µg a.i. difenoconazole mL-1 or greater were required for significant inhibition of mycelial growth. It therefore appears likely that there is currently some control of tan spot as a result of the use of azoxystrobin and to a lesser extent, difenoconazole, for the control of other diseases.

Distribution and eradication of an exotic tephritid fruit fly in Australia: Relevance of invasion theory

Data from the eradication of the incursion of Bactrocera papayae Drew and Hancock (Dipt.: Tephritidae) in Australia (1995-1998) are used to assess the significance of various aspects of invasion theory, including the influence of towns on establishment, influence of propagule pressure on the pattern of establishment, and the existence of source-sink dynamics. Because there were no sentinel traps in place, considerable spread had occurred before the eradication campaign started. The distribution of fly density around the epicentre in the town of Cairns and a transect along the main traffic routes to the north and south fitted a Cauchy model with a tail having the same slope as a power model with an exponent of -2.4 extending to 160 km. The Cauchy model indicated that 50% of the flies on the transect would have occurred within 3.2 km of the epicentre, 90% within 13.2 km, and 99% within 60 km. The two major satellites at Mareeba (35 km from the epicentre in Cairns) and Mossman (65 km) were not used for the transect data and had respectively 15 and 30 times the density predicted by the model. The proportion of traps that caught flies (a measure of site occupancy) fell with distance from the epicentre. B. papayae was trapped consistently on only three of the 16 rainforest transects that were surveyed and these were relatively close to urban areas where eradication efforts were intense. Despite there being no eradication effort in the rainforest, the trends to extinction were similar to those in adjacent areas. The strategy of initially concentrating eradication efforts on the core and major satellites while maintaining a quarantine barrier at the airport and the boundaries of the infested area appears to be the key to the containment and rapid eradication of the incursion.