Australian Bat Lyssavirus: Observations of Natural and Experimental Infection in Bats

This conference abstract gives data and conclusions arising from targeted surveillance of wild bats for naturally occuring Australian bat lyssavirus (ABLV) infection and other central nervous system diseases. It also provides data and conclusions arising from experimental infection of 10 Greyheaded flying foxes (Pteropus poliocephalus).

Impact of mirid (Creontiades spp.) (Hemiptera: Miridae) pest management on Helicoverpa spp. (Lepidoptera: Noctuidae) outbreaks: The case for conserving natural enemies

Creontiades spp. (Hemiptera: Miridae) are sucking pests that attack buds, flowers and young pods in mungbeans, Vigna radiata (L.), causing these structures subsequently to abort. If left uncontrolled, mirids can cause 25-50% yield loss. Traditional industry practice has involved prophylactic applications of dimethoate to control mirids at budding and again a week later. The present trial was initiated to highlight the dangers of such a practice, in particular the risk of a subsequent Helicoverpa spp. lepidopteran pest outbreak. A single application of dimethoate halved the population of important natural enemies of Helicoverpa spp., and caused an above-threshold outbreak of Helicoverpa spp. within 11 days. This shows that even a moderate (e.g. 50%) reduction in natural enemies may be sufficient to increase Helicoverpa spp. populations in mungbeans. As a result, prophylactic sprays should not be used for the control of mirids in mungbeans, and dimethoate should be applied only when mirids are above the economic threshold. Indoxacarb was also tested to establish its effect on Helicoverpa spp., mirids and natural enemies. Indoxacarb showed potential for Helicoverpa spp. control and suppression of mirids and had little impact on natural enemies.

Three new Lasiodiplodia spp. from the tropics, recognized based on DNA sequence comparisons and morphology

Botryosphaeria rhodina (anamorph Lasiodiplodia theobromae) is a common endophyte and opportunistic pathogen on more than 500 tree species in the tropics and subtropics. During routine disease surveys of plantations in Australia and Venezuela several isolates differing from L. theobromae were identified and subsequently characterized based upon morphology and ITS and EF1-a nucleotide sequences. These isolates grouped into three strongly supported clades related to but different from the known taxa, B. rhodina and L. gonubiensis, These have been described here as three new species L. venezuelensis sp. nov., L. crassispora sp. nov. and L. rubropurpurea sp. nov. The three could be distinguished easily from each other and the two described species of Lasiodiplodia, thus confirming phylogenetic separations. Furthermore all five Lasiodiplodia spp. now recognized separated from Diplodia spp. and Dothiorella spp. with 100% bootstrap support.

Bats: Important reservoir hosts of emerging viruses

Bats (order Chiroptera, suborders Megachiroptera and Microchiroptera) are abundant, diverse, and geographically widespread. These mammals provide us with resources, but their importance is minimized and many of their populations and species are at risk, even threatened or endangered. Some of their characteristics (food choices, colonial or solitary nature, population structure, ability to fly, seasonal migration and daily movement patterns, torpor and hibernation, life span, roosting behaviors, ability to echolocate, virus susceptibility) make them exquisitely suitable hosts of viruses and other disease agents. Bats of certain species are well recognized as being capable of transmitting rabies virus, but recent observations of outbreaks and epidemics of newly recognized human and livestock diseases caused by viruses transmitted by various megachiropteran and microchiropteran bats have drawn attention anew to these remarkable mammals. This paper summarizes information regarding chiropteran characteristics and information regarding 66 viruses that have been isolated from bats. From these summaries, it is clear that we do not know enough about bat biology, that we are doing too little in terms of bat conservation, and that there remain a multitude of questions regarding the role of bats in disease emergence.

Detection of Arcobacter spp. in piggery effluent and effluent-irrigated soils in southeast Queensland

Aims: To investigate the occurrence and levels of Arcobacter spp. in pig effluent ponds and effluent-treated soil. Methods and Results: A Most Probable Number (MPN) method was developed to assess the levels of Arcobacter spp. in seven pig effluent ponds and six effluent-treated soils, immediately after effluent irrigation. Arcobacter spp. levels in the effluent ponds varied from 6.5 × 105 to 1.1 × 108 MPN 100 ml-1 and in freshly irrigated soils from 9.5 × 102 to 2.8 × 104 MPN g-1 in all piggery environments tested. Eighty-three Arcobacter isolates were subjected to an abbreviated phenotypic test scheme and examined using a multiplex polymerase chain reaction (PCR). The PCR identified 35% of these isolates as Arcobacter butzleri, 49% as Arcobacter cryaerophilus while 16% gave no band. All 13 nonreactive isolates were subjected to partial 16S rDNA sequencing and showed a high similarity (>99%) to Arcobacter cibarius. Conclusions: A. butzleri, A. cryaerophilus and A. cibarius were isolated from both piggery effluent and effluent-irrigated soil, at levels suggestive of good survival in the effluent pond. Significance and Impact of the Study: This is the first study to provide quantitative information on Arcobacter spp. levels in piggery effluent and to associate A. cibarius with pigs and piggery effluent environments.

Resistance of selected strawberry cultivars to root-knot nematode species (Meloidogyne spp.)

Strawberry runner production areas in Queensland are assessed for the presence of Pratylenchus vulnus (lesion nematode) and Meloidogyne hapla (root-knot nematode) as part of the approval process for sites used in runner production under the approved runner scheme. M. hapla is known to infest strawberry. The ability of three other Meloidogyne species occurring in Queensland to infest this host was investigated. The species M. arenaria, M. incognita and M. javanica, in addition to M. hapla, were able to reproduce on strawberry roots of the cultivar 'Joy', which sustained higher nematode reproduction rates than 'Jewel' and 'Sweet Charlie'. The ability of species other than M. hapla to infest strawberry needs to be recognised in site selection for runner production, and in screening cultivars for resistance to nematodes.

Evolutionary relationships between bat coronaviruses and their hosts

Recent studies have suggested that bats are the natural reservoir of a range of coronaviruses (CoVs), and that rhinolophid bats harbor viruses closely related to the severe acute respiratory syndrome (SARS) CoV, which caused an outbreak of respiratory illness in humans during 2002-2003. We examined the evolutionary relationships between bat CoVs and their hosts by using sequence data of the virus RNA-dependent RNA polymerase gene and the bat cytochrome b gene. Phylogenetic analyses showed multiple incongruent associations between the phylogenies of rhinolophid bats and their CoVs, which suggested that host shifts have occurred in the recent evolutionary history of this group. These shifts may be due to either virus biologic traits or host behavioral traits. This finding has implications for the emergence of SARS and for the potential future emergence of SARS-CoVs or related viruses.

Utilizing the assassin bug, Pristhesancus plagipennis (Hemiptera: Reduviidae), as a biological control agent within an integrated pest management programme for Helicoverpa spp. (Lepidoptera: Noctuidae) and Creontiades spp. (Hemiptera: Miridae) in cotton

Helicoverpa spp. and mirids, Creontiades spp., have been difficult to control biologically in cotton due to their unpredictable temporal abundance combined with a cropping environment often made hostile by frequent usage of broad spectrum insecticides. To address this problem, a range of new generation insecticides registered for use in cotton were tested for compatibility with the assassin bug, Pristhesancus plagipennis (Walker), a potential biological control agent for Helicoverpa spp. and Creontiades spp. Indoxacarb, pyriproxifen, buprofezin, spinosad and fipronil were found to be of low to moderate toxicity on P. plagipennis whilst emamectin benzoate, abamectin, diafenthiuron, imidacloprid and omethaote were moderate to highly toxic. Inundative releases of P. plagipennis integrated with insecticides identified as being of low toxicity were then tested and compared with treatments of P. plagipennis and the compatible insecticides used alone, conventionally sprayed usage practice and an untreated control during two field experiments in cotton. The biological control provided by P. plagipennis nymphs when combined with compatible insecticides provided significant (P<0.001) reductions in Helicoverpa and Creontiades spp. on cotton and provided equivalent yields to conventionally sprayed cotton with half of the synthetic insecticide input. Despite this, the utilization of P. plagipennis in cotton as part of an integrated pest management programme remains unlikely due to high inundative release costs relative to other control technologies such as insecticides and transgenic (Bt) cotton varieties.

Resistance of Brassicaceae plants to root-knot nematode (Meloidogyne spp.) in northern Australia

Brassicaceae plants have the potential as part of an integrated approach to replace fumigant nematicides, providing the biofumigation response following their incorporation is not offset by reproduction of plant-parasitic nematodes on their roots. Forty-three Brassicaceae cultivars were screened in a pot trial for their ability to reduce reproduction of three root-knot nematode isolates from north Queensland, Australia: M. arenaria (NQ1), M. javanica (NQ2) and M. arenaria race 2 (NQ5/7). No cultivar was found to consistently reduce nematode reproduction relative to forage sorghum, the current industry standard, although a commercial fodder radish (Raphanus sativus) and a white mustard (Sinapis alba) line were consistently as resistant to the formation of galls as forage sorghum. A second pot trial screened five commercially available Brassicaceae cultivars, selected for their biofumigation potential, for resistance to two nematode species, M. javanica (NQ2) and M. arenaria (NQ5/7). The fodder radish cv. Weedcheck, was found to be as resistant as forage sorghum to nematode reproduction. A multivariate cluster analysis using the resistance measurements, gall index, nematode number per g of root and multiplication for two nematode species (NQ2 and NQ5/7) confirmed the similarity in resistance between the radish cultivar and forage sorghum. A field trial confirmed the resistance of the fodder radish cv. Weedcheck, with a similar reduction in the number of Meloidogyne spp. juveniles recovered from the roots 8 weeks after planting. The use of fodder radish cultivars as biofumigation crops to manage root-knot nematodes in tropical vegetable production systems deserves further investigation.

The influence of planting density on the production of 'Goldfinger' (Musa spp., AAAB) in the subtropics

'Goldfinger', a tetraploid banana produced from the Fundación Hondureña de Investigación Agrícola (FHIA) breeding program, was released to the Australian industry in 1995. It was promoted as an apple-flavoured dessert banana with resistance to Fusarium wilt race 1 and subtropical race 4, as well as resistance to black and yellow Sigatoka (Mycosphaerella fijiensis and M. musicola, respectively). This study was initiated to provide agronomic information to the banana industry, which was under threat from Fusarium wilt, on a new cultivar which could replace 'Williams' (AAA, Cavendish subgroup) or 'Lady Finger' (AAB, Pome subgroup) in those areas affected by Fusarium wilt. Also few studies had reported on the production characteristics of the new tetraploid hybrids, especially from subtropical areas, and therefore two field sites, one a steep-land farm and the other a level, more productive site, were selected for planting density and spatial arrangement treatments. The optimum density in terms of commercial production, taking into account bunch weight, finger size, length of the production cycle, plant height and ease of management, was 1680 plants/ha on the steep-land site where plants were planted in single rows with 2.5 m × 2.5 m spacings. However on the level site a double-row triangular layout with inter-row distances of 4.5 m to allow vehicular access (1724 plants/ha) gave the best results. With this arrangement plants were in an alternate, triangular arrangement along a row and a spacing of 1.5 m between plants at the points of each triangle and between each block of triangles.

Survival of Campylobacter spp. in darkling beetles (Alphitobius diaperinus) and their larvae in Australia

Campylobacter infection is the most frequently reported notifiable food-borne disease in humans in Australia. Our studies investigated the persistence of Campylobacter spp. in or on darkling beetles (Alphitobius diaperinus) and their larvae. Our results in analyses with chickens confirm that, unless very short turnaround times are used (<72 h), beetles colonized in one production cycle (i.e., one batch of chickens) are most unlikely to still be colonized during the next cycle of chickens.

Review of bats and SARS

Bats have been identified as a natural reservoir for an increasing number of emerging zoonotic viruses, including henipaviruses and variants of rabies viruses. Recently, we and another group independently identified several horse-shoe bat species (genus Rhinolophus) as the reservoir host for a large number of viruses that have a close genetic relationship with the coronavirus associated with severe acute respiratory syndrome (SARS). Our current research focused on the identification of the reservoir species for the progenitor virus of the SARS coronaviruses responsible for outbreaks during 2002-2003 and 2003-2004. In addition to SARS-like coronaviruses, many other novel bat coronaviruses, which belong to groups 1 and 2 of the 3 existing coronavirus groups, have been detected by PCR. The discovery of bat SARS-like coronaviruses and the great genetic diversity of coronaviruses in bats have shed new light on the origin and transmission of SARS coronaviruses.

Infection with Menangle virus in flying foxes (Pteropus spp.) in Australia

Objective: To examine flying foxes (Pteropus spp.) for evidence of infection with Menangle virus. Design: Clustered non-random sampling for serology, virus isolation and electron microscopy (EM). Procedure: Serum samples were collected from 306 Pteropus spp. in northern and eastern Australia and tested for antibodies against Menangle virus (MenV) using a virus neutralisation test (VNT). Virus isolation was attempted from tissues and faeces collected from 215 Pteropus spp. in New South Wales. Faecal samples from 68 individual Pteropus spp. and four pools of faeces were examined by transmission EM following routine negative staining and immunogold labelling. Results: Neutralising antibodies (VNT titres ≥ 8) against MenV were detected in 46% of black flying foxes (P. alecto), 41% of grey-headed flying foxes (P. poliocephalus), 25% of spectacled flying foxes (P. conspicillatus) and 1% of little red flying foxes (P. scapulatus) in Australia. Positive sera included samples collected from P. poliocephalus in a colony adjacent to a piggery that had experienced reproductive disease caused by MenV. Virus-like particles were observed by EM in faeces from Pteropus spp. and reactivity was detected in pooled faeces and urine by immunogold EM using sera from sows that had been exposed to MenV. Attempts to isolate the virus from the faeces and tissues from Pteropus spp. were unsuccessful. Conclusion: Serological evidence of infection with MenV was detected in Pteropus spp. in Australia. Although virus-like particles were detected in faeces, no viruses were isolated from faeces, urine or tissues of Pteropus spp.

Evaluating new insecticides for the control of Helicoverpa spp. (Lepidoptera: Noctuidae) on capsicum and zucchini

The efficacy of insecticides in controlling Helicoverpa spp., predominantly H. armigera (Hubner), on capsicum and zucchini was tested in small plot trials. Indoxacarb, methoxyfenozide, spinosad, emamectin benzoate and novaluron provided control, as measured by the percentage of damaged fruit, equal to or better than standard treatments of methomyl or methomyl alternated with methamidophos on capsicum. The Helicoverpa nucleopolyhedrovirus gave control equivalent to the standard treatment, as did Bacillus thuringiensis aizawai, but B. thuringiensis kurstaki was ineffective. Helicoverpa armigera larvae were present in zucchini flowers but did little damage to the fruit. None of the insecticides significantly reduced the percentage of damaged zucchini fruit compared with the untreated control. Bifenthrin, spinosad, emamectin benzoate and methoxyfenozide were effective in controlling larvae in flowers, while methomyl, B. thuringiensis aizawai, B. thuringiensis kurstaki and novaluron were not effective. Data indicated that all the insecticides effectively controlled larvae of Diaphania indica (Saunders), cucumber moth, in the zucchini flowers. There has been a limited range of insecticides available to manage Helicoverpa spp. in these vegetable crops, but these trials demonstrate the effectiveness of a number of newer insecticides that could be used and that would be compatible with integrated pest management programs in the crops.

Oidiopsis (Erysiphaceae) on Euphorbia spp. in Australia and Vanuatu

One specimen of powdery mildew on Euphorbia cyathophora from Vanuatu and 11 specimens on E. cyathophora, E. dentata, E. heterophylla and E. leucocephala from Australia were studied. All were shown to represent the Oidiopsis anamorph of Leveillula taurica, which is described. This is the first record of Oidiopsis in Vanuatu. E. leucocephala is a new host record for this powdery mildew.