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

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

Characterization of north-eastern Australian environments using APSIM for increasing rainfed maize production

Recurring water stresses are a major risk factor for rainfed maize cropping across the highly diverse agro-ecological environments of Queensland (Qld) and northern New South Wales (NNSW). Enhanced understanding of such agro-ecological diversity is necessary to more consistently sample target production environments for testing and targeting release of improved germplasm, and to improve the efficiency of the maize pre-breeding and breeding programs of Qld and New South Wales. Here, we used the Agricultural Production Systems Simulator (APSIM) – a well validated maize crop model to characterize the key distinctive water stress patterns and risk to production across the main maize growing regions of Qld and NNSW located between 15.8° and 31.5°S, and 144.5° and 151.8°E. APSIM was configured to simulate daily water supply demand ratios (SDRs) around anthesis as an indicator of the degree of water stress, and the final grain yield. Simulations were performed using daily climatic records during the period between 1890 and 2010 for 32 sites-soils in the target production regions. The runs were made assuming adequate nitrogen supply for mid-season maize hybrid Pioneer 3153. Hierarchical complete linkage analyses of the simulated yield resulted in five major clusters showing distinct probability distribution of the expected yields and geographic patterns. The drought stress patterns and their frequencies using SDRs were quantified using multivariate statistical methods. The identified stress patterns included no stress, mid-season (flowering) stress, and three terminal stresses differing in terms of severity. The combined frequency of flowering and terminal stresses was highest (82.9%), mainly in sites-soils combinations in the west of Qld and NNSW. Yield variability across the different sites-soils was significantly related to the variability in frequencies of water stresses. Frequencies of water stresses within each yield cluster tended to be similar, but different across clusters. Sites-soils falling within each yield cluster therefore could be treated as distinct maize production environments for testing and targeting newly developed maize cultivars and hybrids for adaptation to water stress patterns most common to those environments.

Identification of a new Polerovirus (family Luteoviridae) associated with cotton bunchy top disease in Australia

Cotton bunchy top (CBT) disease has caused significant yield losses in Australia and is now managed by control of its vector, the cotton aphid (Aphis gossypii). Its mode of transmission and similarities in symptoms to cotton Blue Disease suggested it may also be caused by a luteovirus or related virus. Degenerate primers to conserved regions of the genomes of the family Luteoviridae were used to amplify viral cDNAs from CBT-affected cotton leaf tissue that were not present in healthy plants. Partial genome sequence of a new virus (Cotton bunchy top virus, CBTV) was obtained spanning part of the RNA-dependent-RNA-polymerase (RdRP), all of the coat protein and part of the aphid-transmission protein. CBTV sequences could be detected in viruliferous aphids able to transmit CBT, but not aphids from non-symptomatic plants, indicating that it is associated with the disease and may be the causal agent. All CBTV open-reading frames had their closest similarity to viruses of the genus Polerovirus. The partial RdRP had 90 % amino acid identity to the RdRP of Cotton leafroll dwarf virus (CLRDV) that causes cotton blue disease, while other parts of the genome were more similar to other poleroviruses. The sequence similarity and genome organization of CBTV suggest that it should be considered a new member of the genus Polerovirus. This partial genome sequence of CBTV opens up the possibility for developing diagnostic tests for detection of the virus in cotton plants, aphids and weeds as well as alternative strategies for engineering CBT resistance in cotton plants through biotechnology. © 2012 Australasian Plant Pathology Society Inc.

Morphological and molecular diversity of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C.gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily-primed PCR (ap-PCR) and DNA sequencing studies did not differentiate isolates of C.gloeosporioides from anthracnose and pepper spot lesions. The ap-PCR identified 21 different genotypes of C.gloeosporioides, three of which were predominant. A specific genotype identified using ap-PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and -tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap-PCR, the possibility of a lychee specific group of C.gloeosporioides is discussed.

Development of cat's claw creeper leaf-tying moth Hypocosmia pyrochroma (Lepidoptera: Pyralidae) at different temperatures: Implications for establishment as a biological control agent in Australia and South Africa

The leaf-tying moth Hypocosmia pyrochroma Jones (Lepidoptera: Pyralidae), a native of sub tropical South America, has been introduced as a biological control agent for cat’s claw creeper, Dolichandra unguis-cati (L.) Lohman (Bignoniaceae), in Australia and South Africa. So far there has been no evidence of its field establishment in either country. A narrow temperature tolerance is a potential limiting factor for the establishment of weed biological control insects in novel habitats. In this study, we evaluated the effect of seven constant temperatures (12–40 °C) on the survival and development of H. pyrochroma in temperature-controlled cabinets. Temperatures between 20 and 30 °C were the most favorable for adult survival, oviposition, egg hatching, and larval and pupal development. Adult survival (12–40 °C) and egg development (15–35 °C) showed tolerance for wider temperature ranges than oviposition, and larval and pupal development, which were all negatively affected by both high (>30 °C) and low (<20 °C) temperatures. The degree-day (DD) requirement to complete a generation was estimated as 877 above a threshold temperature of 12 °C. Based on DD requirements and an obligatory winter diapause of pupae from mid-autumn to mid-spring, the potential number of generations (egg to adult) the leaf-tying moth can complete in a year in Australia or South Africa range from one to three. A climate-matching model predicted that the inland regions of both Australia and South Africa are less favorable for H. pyrochroma than the coastal areas. The study suggested that H. pyrochroma is more likely to establish in the coastal areas of Australia where most of the cat’s claw creeper infestations occur, than in South Africa where most of the cat’s claw creeper infestations are inland.

The rph2 Gene Is Responsible for High Level Resistance to Phosphine in Independent Field Strains of Rhyzopertha dominica

The lesser grain borer Rhyzopertha dominica (F.) is one of the most destructive insect pests of stored grain. This pest has been controlled successfully by fumigation with phosphine for the last several decades, though strong resistance to phosphine in many countries has raised concern about the long term usefulness of this control method. Previous genetic analysis of strongly resistant (SR) R. dominica from three widely geographically dispersed regions of Australia, Queensland (SRQLD), New South Wales (SRNSW) and South Australia (SRSA), revealed a resistance allele in the rph1 gene in all three strains. The present study confirms that the rph1 gene contributes to resistance in a fourth strongly resistant strain, SR2(QLD), also from Queensland. The previously described rph2 gene, which interacts synergistically with rph1 gene, confers strong resistance on SRQLD and SRNSW. We now provide strong circumstantial evidence that weak alleles of rph2, together with rph1, contribute to the strong resistance phenotypes of SRSA and SR2(QLD). To test the notion that rph1 and rph2 are solely responsible for the strong resistance phenotype of all resistant R. dominica, we created a strain derived by hybridising the four strongly resistant lines. Following repeated selection for survival at extreme rates of phosphine exposure, we found only slightly enhanced resistance. This suggests that a single sequence of genetic changes was responsible for the development of resistance in these insects.

Differential recognition by tick-resistant cattle of the recombinantly expressed Rhipicephalus microplus serine protease inhibitor-3 (RMS-3)

Rhipicephalus micro plus is an important bovine ectoparasite, widely distributed in tropical and subtropical regions of the world causing large economic losses to the cattle industry. Its success as an ectoparasite is associated with its capacity to disarm the antihemostatic and anti-inflammatory reactions of the host. Serpins are protease inhibitors with an important role in the modulation of host-parasite interactions. The cDNA that encodes for a R. microplus serpin was isolated by RACE and subsequently cloned into the pPICZ alpha A vector. Sequence analysis of the cDNA and predicted amino acid showed that this cDNA has a conserved serpin domain. B- and T-cell epitopes were predicted using bioinformatics tools. The recombinant R. microplus serpin (rRMS-3) was secreted into the culture media of Pichia pastoris after methanol induction at 0.2 mg l(-1) qRT-PCR expression analysis of tissues and life cycle stages demonstrated that RMS-3 was mainly expressed in the salivary glands of female adult ticks. Immunological recognition of the rRMS-3 and predicted B-cell epitopes was tested using tick-resistant and susceptible cattle sera. Only sera from tick-resistant bovines recognized the B-cell epitope AHYNPPPPIEFT (Seq7). The recombinant RMS-3 was expressed in P. pastoris, and ELISA screening also showed higher recognition by tick-resistant bovine sera. The results obtained suggest that RMS-3 is highly and specifically secreted into the bite site of R. microplus feeding on tick-resistant bovines. Capillary feeding of semi-engorged ticks with anti-AHYNPPPPIEFT sheep sera led to an 81.16% reduction in the reproduction capacity of R. microplus. Therefore, it is possible to conclude that R. microplus serpin (RMS-3) has an important role in the host-parasite interaction to overcome the immune responses in resistant cattle. (C) 2012 Elsevier GmbH. All rights reserved.

High resolution mapping of Dense spike-ar (dsp.ar) to the genetic centromere of barley chromosome 7H

Spike density in barley is under the control of several major genes, as documented previously by genetic analysis of a number of morphological mutants. One such class of mutants affects the rachis internode length leading to dense or compact spikes and the underlying genes were designated dense spike (dsp). We previously delimited two introgressed genomic segments on chromosome 3H (21 SNP loci, 35.5 cM) and 7H (17 SNP loci, 20.34 cM) in BW265, a BC7F3 nearly isogenic line (NIL) of cv. Bowman as potentially containing the dense spike mutant locus dsp.ar, by genotyping 1,536 single nucleotide polymorphism (SNP) markers in both BW265 and its recurrent parent. Here, the gene was allocated by high-resolution bi-parental mapping to a 0.37 cM interval between markers SC57808 (Hv_SPL14)-CAPSK06413 residing on the short and long arm at the genetic centromere of chromosome 7H, respectively. This region putatively contains more than 800 genes as deduced by comparison with the collinear regions of barley, rice, sorghum and Brachypodium, Classical map-based isolation of the gene dsp.ar thus will be complicated due to the infavorable relationship of genetic to physical distances at the target locus.

The diagnosis and management of a case of leishmaniosis in a dog imported to Australia

This case study discusses in detail for the first time the diagnosis and management of a case of leishmaniosis in a dog imported to Australia. The dog presented with epistaxis and a non-regenerative anaemia five years after being imported from Europe. Protozoa were identified within macrophages in bone marrow and splenic cytology. A Leishmania indirect fluorescent antibody test was performed and was positive while an Ehrlichia canis antibody test was negative. Polymerase chain reaction of the ITS-1 and ITS-2 regions of skin, lymph node, spleen and bone marrow were all positive for Leishmania infantum. The dog was treated with amphotericin B with a strong clinical response. The importance of thorough diagnostics in non-endemic areas, particularly Australia, is discussed. Treatment with amphotericin B is discussed. Vigilance, disease reporting and response frameworks are recommended for non-endemic areas. © 2014 Elsevier B.V.

Development of cat’s claw creeper leaf-tying moth Hypocosmia pyrochroma (Lepidoptera: Pyralidae) at different temperatures: Implications for establishment as a biological control agent in Australia and South Africa

The leaf-tying moth Hypocosmia pyrochroma Jones (Lepidoptera: Pyralidae), a native of sub tropical South America, has been introduced as a biological control agent for cat’s claw creeper, Dolichandra unguis-cati (L.) Lohman (Bignoniaceae), in Australia and South Africa. So far there has been no evidence of its field establishment in either country. A narrow temperature tolerance is a potential limiting factor for the establishment of weed biological control insects in novel habitats. In this study, we evaluated the effect of seven constant temperatures (12–40 °C) on the survival and development of H. pyrochroma in temperature-controlled cabinets. Temperatures between 20 and 30 °C were the most favorable for adult survival, oviposition, egg hatching, and larval and pupal development. Adult survival (12–40 °C) and egg development (15–35 °C) showed tolerance for wider temperature ranges than oviposition, and larval and pupal development, which were all negatively affected by both high (>30 °C) and low (<20 °C) temperatures. The degree-day (DD) requirement to complete a generation was estimated as 877 above a threshold temperature of 12 °C. Based on DD requirements and an obligatory winter diapause of pupae from mid-autumn to mid-spring, the potential number of generations (egg to adult) the leaf-tying moth can complete in a year in Australia or South Africa range from one to three. A climate-matching model predicted that the inland regions of both Australia and South Africa are less favorable for H. pyrochroma than the coastal areas. The study suggested that H. pyrochroma is more likely to establish in the coastal areas of Australia where most of the cat’s claw creeper infestations occur, than in South Africa where most of the cat’s claw creeper infestations are inland.

Effect of Increasing Low Dose Simplexin Exposure in Cattle consuming Pimelia trychostachya

Pimelea species (or desert riceflower) are small native plants endemic to the drier inland pastoral regions of Australia, which cause a unique syndrome in grazing cattle characterised by submandibular oedema and oedema in the brisket area as a result of right-sided heart failure attributed to the toxin simplexin. Field evidence suggests that poisoning can occur through minor, inadvertent consumption of Pimelea plant material, but the minimum simplexin intake required to induce Pimelea poisoning is not known. In this study, mild Pimelea poisoning was induced at a daily dose of 12.5 mg Pimelea/kg bodyweight per day, equivalent to 2.5 µg simplexin/kg bodyweight per day, demonstrating the high potential toxicity of these plant species. Effects in all animals diminished with prolonged low dose feeding and we postulate that these animals developed mechanisms for detoxifying simplexin, 1, possibly through rumen bacteria adaptation or activation of liver enzymes.

Development and testing of allometric equations for estimating above-ground biomass of mixed-species environmental plantings

To quantify the impact that planting indigenous trees and shrubs in mixed communities (environmental plantings) have on net sequestration of carbon and other environmental or commercial benefits, precise and non-biased estimates of biomass are required. Because these plantings consist of several species, estimation of their biomass through allometric relationships is a challenging task. We explored methods to accurately estimate biomass through harvesting 3139 trees and shrubs from 22 plantings, and collating similar datasets from earlier studies, in non-arid (>300mm rainfallyear-1) regions of southern and eastern Australia. Site-and-species specific allometric equations were developed, as were three types of generalised, multi-site, allometric equations based on categories of species and growth-habits: (i) species-specific, (ii) genus and growth-habit, and (iii) universal growth-habit irrespective of genus. Biomass was measured at plot level at eight contrasting sites to test the accuracy of prediction of tonnes dry matter of above-ground biomass per hectare using different classes of allometric equations. A finer-scale analysis tested performance of these at an individual-tree level across a wider range of sites. Although the percentage error in prediction could be high at a given site (up to 45%), it was relatively low (<11%) when generalised allometry-predictions of biomass was used to make regional- or estate-level estimates across a range of sites. Precision, and thus accuracy, increased slightly with the level of specificity of allometry. Inclusion of site-specific factors in generic equations increased efficiency of prediction of above-ground biomass by as much as 8%. Site-and-species-specific equations are the most accurate for site-based predictions. Generic allometric equations developed here, particularly the generic species-specific equations, can be confidently applied to provide regional- or estate-level estimates of above-ground biomass and carbon. © 2013 Elsevier B.V.

Molecular phylogenetic analysis reveals six new species of Diaporthe from Australia

Six new species of Diaporthe, D. beilharziae on Indigofera australis, D. fraxini-angustifoliae on Fraxinus angustifolia subsp. oxycarpa, D. litchicola on Litchi chinensis, D. nothofagi on Nothofagus cunninghamii, D. pascoei on Persea americana and D. salicicola on Salix purpurea from Australia are described and illustrated based on morphological characteristics and molecular analyses. Three of the new species no longer produced sporulating structures in culture and two of these were morphologically described from voucher specimens. Phylogenetic relationships of the new species with other Diaporthe species are revealed by DNA sequence analyses based on the internal transcribed spacer (ITS) region, and partial regions of the β-tubulin (BT) and translation elongation factor 1-alpha (TEF). © 2013 Mushroom Research Foundation.

Population dynamics of Thaumastocoris peregrinus in Eucalyptus plantations of South Africa

Thaumastocoris peregrinus is a sap-sucking insect that infests non-native Eucalyptus plantations in Africa, New Zealand, South America and parts of Southern Europe, in addition to street trees in parts of its native range of Australia. In South Africa, pronounced fluctuations in the population densities have been observed. To characterise spatiotemporal variability in T. peregrinus abundance and the factors that might influence it, we monitored adult population densities at six sites in the main eucalypt growing regions of South Africa. At each site, twenty yellow sticky traps were monitored weekly for 30 months, together with climatic data. We also characterised the influence of temperature on growth and survival experimentally and used this to model how temperature may influence population dynamics. T. peregrinus was present throughout the year at all sites, with annual site-specific peaks in abundance. Peaks occurred during autumn (February-April) for the Pretoria site, summer (November-January) for the Zululand site and spring (August-October) for the Tzaneen, Sabie and Piet Retief monitoring sites. Temperature (both experimental and field-collected), humidity and rainfall were mostly weakly, or not at all, associated with population fluctuations. It is clear that a complex interaction of these and other factors (e.g. host quality) influence population fluctuations in an annual, site specific cycle. The results obtained not only provide insights into the biology of T. peregrinus, but will also be important for future planning of monitoring and control programs using semiochemicals, chemical insecticides or biological control agents. © 2014 Springer-Verlag Berlin Heidelberg.

The systematics of Endoraecium in Australia

Endoraecium (Raveneliaceae, Pucciniales) is a genus of rust that infects several species of Acacia (Fabaceae) in Australia, south-east Asia and Hawaii. Thirteen species of Endoraecium have been described, including seven species that are endemic to Australia, one species to south-east Asia and five to Hawaii. This study investigated the systematics of Endoraecium from 50 specimens in Australia and south-east Asia with a combined morphological and molecular approach. Phylogenetic analyses were conducted on combined datasets of the SSU, ITS and LSU regions of rDNA. The recovered phylogeny (i) supported a recent division of Endoraecium digitatum into five separate species based on morphology and host specificity and (ii) found lineages that did not correspond with known species.