Field and laboratory evaluations of fungicides for the control of brown spot (Corynespora cassiicola) and black spot (Asperisporium caricae) of papaya in far north Queensland, Australia

Several chemicals including strobilurins (pyraclostrobin and azoxystrobin), triazoles (difenoconazole and tebuconazole), dithiocarbamates (propineb, metiram, ziram and mancozeb) and the phthalimide chlorothalonil were evaluated in three field experiments in north Queensland, Australia, for the control of brown spot (caused by Corynespora cassiicola) and black spot (caused by Asperisporium caricae) of papaya. Chlorothalonil and pyraclostrobin were shown to be more effective than the industry standard, mancozeb, for the control of brown spot. In the black spot experiments, difenoconazole, pyraclostrobin and chlorothalonil used alone or in spray programs were as effective as, or better than, the industry standards, mancozeb and tebuconazole. Plants treated with pyraclostrobin and difenoconazole had more fruit unaffected by black spot (97% and 99% respectively) than plants treated with tebuconazole (51%), mancozeb (20%) and the untreated controls (1%). Laboratory tests also showed that A. caricae was more sensitive to difenoconazole (EC50 of 2ppm) than tebuconazole (EC50 of 14ppm). In 2007, off-label permits were obtained for chlorothalonil for control of brown spot and difenoconazole and chlorothalonil for the control of black spot of papaya.

Field isolates of Tomato spotted wilt virus overcoming resistance in capsicum in Australia

In 2002 at Virginia, South Australia, capsicum cultivars having the Tsw resistance gene against Tomato spotted wilt virus (TSWV) developed symptoms typical of TSWV infection and several glasshouse-grown crops were almost 100% infected. Samples reacted with TSWV antibodies in ELISA. Virus isolates from infected plants induced severe systemic symptoms, rather than a hypersensitive reaction, when inoculated onto capsicum cultivars and Capsicum chinense genotypes ( PI 152225 and PI 159236) that carry the Tsw resistance gene. Isolates virulent towards the Tsw gene had molecular and biological properties very similar to standard TSWV isolates, including a hypersensitive reaction in Sw-5 (TSWV-resistant) tomato genotypes. Tsw-virulent isolates were found during surveys at Virginia in 2002 and 2004 in both TSWV-resistant and susceptible cultivars of capsicum and tomato.

Identification of two distinct bovine parainfluenza virus type 3 genotypes

The partial gene sequencing of the matrix (M) protein from seven clinical isolates of bovine parainfluenza virus type 3 (BPIV-3), and the complete sequencing of a representative isolate (Q5592) was completed in this study. Nucleotide sequence analysis was initiated because of the failure of in-house BPIV-3 RT-PCR methods to yield expected products for four of the isolates. Phylogenetic reconstructions based on the nucleotide sequences for the M-protein and the entire genome, using all of the available BPIV-3 nucleotide sequences, demonstrated that there were two distinct BPIV-3 genotypes (BPIV-3a and BPIV-3b). These newly identified genotypes have implications for the development of BPIV-3 molecular detection methods and may also impact on BPIV-3 vaccine formulations.

Responses in carcass lean pH at 24 hours post-mortem in lines of Large White pigs selected for growth rate on a fixed ration over a set time

A restricted maximum likelihood analysis applied to an animal model showed no significant differences (P > 0.05) in pH value of the longissimus dorsi measured at 24 h post-mortem (pH24) between high and low lines of Large White pigs selected over 4 years for post-weaning growth rate on restricted feeding. Genetic and phenotypic correlations between pH24 and production and carcass traits were estimated using all performance testing records combined with the pH24 measurements (5.05-7.02) on slaughtered animals. The estimate of heritability for pH24 was moderate (0.29 ± 0.18). Genetic correlations between pH24 and production or carcass composition traits, except for ultrasonic backfat (UBF), were not significantly different from zero. UBF had a moderate, positive genetic correlation with pH24 (0.24 ± 0.33). These estimates of genetic correlations affirmed that selection for increased growth rate on restricted feeding is likely to result in limited changes in pH24 and pork quality since the selection does not put a high emphasis on reduced fatness.

SSR analysis of cultivated groundnut (Arachis hypogaea L.) germplasm resistant to rust and late leaf spot diseases

Cultivated groundnut (Arachis hypogaea L.) is an agronomically and economically important oilseed crop grown extensively throughout the semi-arid tropics of Asia, Africa and Latin America. Rust (Puccinia arachidis) and late leaf spot (LLS, Phaseoisariopsis personata) are among the major diseases causing significant yield loss in groundnut. The development of varieties with high levels of resistance has been constrained by adaptation of disease isolates to resistance sources and incomplete resistance in resistant sources. Despite the wide range of morphological diversity observed in the cultivated groundnut gene pool, molecular marker analyses have thus far been unable to detect a parallel level of genetic diversity. However, the recent development of simple sequence repeat (SSR) markers presents new opportunities for molecular diversity analysis of cultivate groundnut. The current study was conducted to identify diverse disease resistant germplasm for the development of mapping populations and for their introduction into breeding programs. Twenty-three SSRs were screened across 22 groundnut genotypes with differing levels of resistance to rust and LLS. Overall, 135 alleles across 23 loci were observed in the 22 genotypes screened. Twelve of the 23 SSRs (52%) showed a high level of polymorphism, with PIC values ≥0.5. This is the first report detecting such high levels of genetic polymorphism in cultivated groundnut. Multi-dimensional scaling and cluster analyses revealed three well-separated groups of genotypes. Locus by locus AMOVA and Kruskal-Wallis one-way ANOVA identified candidate SSR loci that may be valuable for mapping rust and LLS resistance. The molecular diversity analysis presented here provides valuable information for groundnut breeders designing strategies for incorporating and pyramiding rust and late leaf spot resistances and for molecular biologists wishing to create recombinant inbred line populations to map these traits.

Absolute quantitation of Marek's disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR

The further development of Taqman quantitative real-time PCR (qPCR) assays for the absolute quantitation of Marek's disease virus serotype 1 (MDV1) and Herpesvirus of turkeys (HVT) viruses is described and the sensitivity and reproducibility of each assay reported. Using plasmid DNA copies, the lower limit of detection was determined to be 5 copies for the MDV1 assay and 75 copies for the HVT assay. Both assays were found to be highly reproducible for Ct values and calculated copy numbers with mean intra- and inter-assay coefficients of variation being less than 5% for Ct and 20% for calculated copy number. The genome copy number of MDV1 and HVT viruses was quantified in PBL and feather tips from experimentally infected chickens, and field poultry dust samples. Parallelism was demonstrated between the plasmid-based standard curves, and standard curves derived from infected spleen material containing both viral and host DNA, allowing the latter to be used for absolute quantification. These methods should prove useful for the reliable differentiation and absolute quantitation of MDV1 and HVT viruses in a wide range of samples.

Cucumber mosaic virus infection of kava (Piper methysticum) and implications for cultural control of kava dieback disease

Cucumber mosaic virus (CMV) was found by reverse transcription polymerase chain reaction (RT-PCR) to be not fully systemic in naturally infected kava (Piper methysticum) plants in Fiji. Twenty-six of 48 samples (54%) from various tissues of three recently infected plants were CMV-positive compared with 7/51 samples (14%) from three long-term infections (plants affected by dieback for more than 1 year). The virus was also found to have a limited ability to move into newly formed stems. CMV was detected in only 2/23 samples taken from re-growth stems arising from known CMV infected/dieback affected plants. Mechanical inoculation experiments conducted in Fiji indicate that the known kava intercrop plants banana (Musa spp.), pineapple (Ananas comosus), peanut (Arachis hypogaea) and the common weed Mikania micrantha are potential hosts for a dieback-causing strain of CMV It was not possible to transmit the virus mechanically to the common kava intercrop plants taro (Colocasia esculenta), Xanthosoma sp., sweet potato (Ipomoea batatas), yam (Dioscorea alata), papaya (Carica papaya) or the weed Momordica charantia. Implications of the results of this research on a possible integrated disease management strategy are discussed.

Capsicum chlorosis virus infecting Capsicum annuum in the East Kimberley region of Western Australia

Capsicum chlorosis virus (CaCV) was detected in field grown Capsicum annuum from Kununurra in northeast Western Australia. Identification of the Kununurra isolate (WA-99) was confirmed using sap transmission to indicator hosts, positive reactions with tospovirus serogroup IV-specific antibodies and CaCV-specific primers, and amino acid sequence comparisons that showed >97% identity with published CaCV nucleocapsid gene sequences. The reactions of indicator hosts to infection with WA-99 often differed from those of the type isolate from Queensland. The virus multiplied best when test plants were grown at warm temperatures. CaCV was not detected in samples collected in a survey of C. annuum crops planted in the Perth Metropolitan area.

Comparative productivity of irrigated short-term ryegrass (Lolium multiflorum) pasture receiving nitrogen, grown alone or in a mixture with white (Trifolium repens) and Persian (T. resupinatum) clovers

Dairy farms in subtropical Australia use irrigated, annually sown short-term ryegrass (Lolium multiflorum) or mixtures of short-term ryegrass and white (Trifolium repens) and Persian (shaftal) (T. resupinatum) clover during the winter-spring period in all-year-round milk production systems. A series of small plot cutting experiments was conducted in 3 dairying regions (tropical upland, north Queensland, and subtropical southeast Queensland and northern New South Wales) to determine the most effective rate and frequency of application of nitrogen (N) fertiliser. The experiments were not grazed, nor was harvested material returned to the plots, after sampling. Rates up to 100 kg N/ha.month (as urea or calcium ammonium nitrate) and up to 200 kg N/ha every 2 months (as urea) were applied to pure stands of ryegrass in 1991. In 1993 and 1994, urea, at rates up to 150 kg N/ha.month and to 200 kg N/ha every 2 months, was applied to pure stands of ryegrass; urea, at rates up to 50 kg N/ha.month, was also applied to ryegrass-clover mixtures. The results indicate that applications of 50-85 kg N/ha.month can be recommended for short-term ryegrass pastures throughout the subtropics and tropical uplands of eastern Australia, irrespective of soil type. At this rate, dry matter yields will reach about 90% of their potential, forage nitrogen concentration will be increased, there is minimal risk to stock from nitrate poisoning and there will be no substantial increase in soil N. The rate of N for ryegrass-clover pastures is slightly higher than for pure ryegrass but, at these rates, the clover component will be suppressed. However, increased ryegrass yields and higher forage nitrogen concentrations will compensate for the reduced clover component. At application rates up to 100 kg N/ha.month, build-up of NO3--N and NH4+-N in soil was generally restricted to the surface layers (0-20 cm) of the soil, but there was a substantial increase throughout the soil profile at 150 kg N/ha.month. The build-up of NO3--N and NH4+-N was greater and was found at lower rates on the lighter soil compared with heavy clays. Generally, most of the soil N was in the NO3--N form and most was in the top 20 cm.

Carrot as a natural host of Watermelon mosaic virus

Carrot was confirmed as a new natural and experimental host of Watermelon mosaic virus by serology, host reactions and sequence comparisons of the coat protein.

Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms

Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus, caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10(7) copies mu g(-1) DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10(6)-10(8) copies of CyHV-2 mu g(-1) DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22-10 degrees C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (+/- SE) of 7.3 +/- 11 to 394 +/- 55 mu g(-1) DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease.

Variation in acquisition of Fiji disease virus by Perkinsiella saccharicida (Hemiptera: Delphacidae).

Fiji leaf gall, caused the Fiji disease virus (genus Fijivirus, family Reoviridae, FDV), is a serious disease of sugarcane, Saccharum officinarum L., in Australia and several other Asia-Pacific countries. In Australia FDV is transmitted only by the planthopper Perkinsiella saccharicida Kirkaldy (Hemiptera: Delphacidae), in a propagative manner. Successful transmission of FDV by single planthoppers confined to individual virus free plants is highly variable, even under controlled conditions. The research reported here addresses two possible sources of this variation: 1) gender, wing form, and life stage of the planthopper; and 2) genotype of the source plant. The acquisition of FDV by macropterous males, macropterous females, brachypterous females, and nymphs of P. saccharicida from infected plants was investigated using reverse transcription-polymerase chain reaction to diagnose FDV infection in the vector. The proportion of individuals infected with FDV was not statistically related to life stage, gender, or adult wing form of the vector. The acquisition of FDV by P. saccharicida from four cultivars of sugarcane was compared to assess the influence of plant genotype on acquisition. Those planthopper populations reared on diseased 'NCo310' plants had twice as many infected planthoppers as those reared on 'Q110', 'WD1', and 'WD2'. Therefore, variation in FDV acquisition in this system is not the result of variation in the gender, wing form and life stage of the P. saccharicida vectors. The cultivar used as the source plant to rear vector populations does affect the proportion of infected planthoppers in a population.

Bats and Emerging Zoonoses: Henipaviruses and SARS.

Nearly 75% of all emerging infectious diseases (EIDs) that impact or threaten human health are zoonotic. The majority have spilled from wildlife reservoirs, either directly to humans or via domestic animals. The emergence of many can be attributed to predisposing factors such as global travel, trade, agricultural expansion, deforestation habitat fragmentation, and urbanization; such factors increase the interface and or the rate of contact between human, domestic animal, and wildlife populations, thereby creating increased opportunities for spillover events to occur. Infectious disease emergence can be regarded as primarily an ecological process. The epidemiological investigation of EIDs associated with wildlife requires a trans-disciplinary approach that includes an understanding of the ecology of the wildlife species, and an understanding of human behaviours that increase risk of exposure. Investigations of the emergence of Nipah virus in Malaysia in 1999 and severe acute respiratory syndrome (SARS) in China in 2003 provide useful case studies. The emergence of Nipah virus was associated with the increased size and density of commercial pig farms and their encroachment into forested areas. The movement of pigs for sale and slaughter in turn led to the rapid spread of infection to southern peninsular Malaysia, where the high-density, largely urban pig populations facilitated transmission to humans. Identifying the factors associated with the emergence of SARS in southern China requires an understanding of the ecology of infection both in the natural reservoir and in secondary market reservoir species. A necessary extension of understanding the ecology of the reservoir is an understanding of the trade, and of the social and cultural context of wildlife consumption. Emerging infectious diseases originating from wildlife populations will continue to threaten public health. Mitigating and managing the risk requires an appreciation of the connectedness between human, livestock and wildlife health, and of the factors and processes that disrupt the balance.

Should the 40-year-old practice of releasing virulent myxoma virus to control rabbits (Oryctolagus cuniculus) be continued?

Release of virulent myxoma virus has been a key component of rabbit-control operations in Queensland, Australia, since the 1960s but its use rests on anecdotal reports. During a routine operation to release virulent myxoma virus we found no evidence to support the continued regular use of the technique in south-west Queensland. Radio-tagged rabbits inoculated with virulent myxoma virus contracted the disease but failed to pass enough virus to other rabbits to spread the disease. Rabbits with clinical signs of myxomatosis that were shot were infected with field strain derived from the original laboratory strain released in 1950 rather than the virulent strain that has been released annually. There was no change in rabbit survival or abundance caused by the release. Nevertheless, the release of virulent virus may be useful against isolated pockets of rabbits mainly because field strains are less likely to be present. Such pockets are more common now that rabbit haemorrhagic disease virus is established in Queensland.

Susceptibility of source plants to sugarcane Fiji disease virus influences the acquisition and transmission of the virus by the planthopper vector Perkinsiella saccharicida

Fiji leaf gall (FLG) caused by Sugarcane Fiji disease virus (SCFDV) is transmitted by the planthopper Perkinsiella saccharicida. FLG is managed through the identification and exploitation of plant resistance. The glasshouse-based resistance screening produced inconsistent transmission results and the factors responsible for that are not known. A series of glasshouse trials conducted over a 2-year period was compared to identify the factors responsible for the erratic transmission results. SCFDV transmission was greater when the virus was acquired by the vector from a cultivar that was susceptible to the virus than when the virus was acquired from a resistant cultivar. Virus acquisition by the vector was also greater when the vector was exposed to the susceptible cultivars than when exposed to the resistant cultivar. Results suggest that the variation in transmission levels is due to variation in susceptibility of sugarcane cultivars to SCFDV used for virus acquisition by the vector.