Field cases of aflatoxicosis in pigs

Five cases of aflatoxicosis in pigs in southern Queensland are described. One peracute case where aflatoxin concentrations of up to 5000pg aflatoxin B,/kg were demonstrated in stomach contents was presumed to be caused by consumption of mouldy bread. High levels of toxins were also present in the livers. Two cases of acute toxicity were caused by feeding mouldy peanut screenings containing 22000~9 aflatoxin B,/kg. One case of subacute and one of chronic toxicity were caused by sorghum grain based rations with lower aflatoxin levels (4640 and 255 pg/kg). Peracute toxicity caused collapse and deaths within several hours, acute toxicity caused deaths within 12 h and with subacute toxicity deaths occured after 3 weeks on a toxic ration. Anorexia and ill thrift affecting only growing animals were seen with chronic toxicity. Extensive centrilobular liver necrosis and haemorrhage occured with peracute toxicity and in cases of acute poisoning there was hepatic centrilobular cellular infiltration, hepatocyte swelling and bile stasis. With subacute toxicity hepatocyte vacuolation together with bile stasis and bile ductule hyperplasia were seen.

Piggery pond sludge as a nitrogen source for crops. 1. Mineral N supply estimated from laboratory incubations and field application of stockpiled and wet sludge

Piggery pond sludge (PPS) was applied, as-collected (Wet PPS) and following stockpiling for 12 months (Stockpiled PPS), to a sandy Sodosol and clay Vertosol at sites on the Darling Downs of Queensland. Laboratory measures of N availability were carried out on unamended and PPS-amended soils to investigate their value in estimating supplementary N needs of crops in Australia's northern grains region. Cumulative net N mineralised from the long-term (30 weeks) leached aerobic incubation was described by a first-order single exponential model. The mineralisation rate constant (0.057/week) was not significantly different between Control and PPS treatments or across soil types, when the amounts of initial mineral N applied in PPS treatments were excluded. Potentially mineralisable N (No) was significantly increased by the application of Wet PPS, and increased with increasing rate of application. Application of Wet PPS significantly increased the total amount of inorganic N leached compared with the Control treatments. Mineral N applied in Wet PPS contributed as much to the total mineral N status of the soil as did that which mineralised over time from organic N. Rates of C02 evolution during 30 weeks of aerobic leached incubation indicated that the Stockpiled PPS was more stabilised (19-28% of applied organic C mineralised) than the WetPPS (35-58% of applied organic C mineralised), due to higher lignin content in the former. Net nitrate-N produced following 12 weeks of aerobic non-leached incubation was highly correlated with net nitrate-N leached during 12 weeks of aerobic incubation (R^2 = 0.96), although it was <60% of the latter in both sandy and clayey soils. Anaerobically mineralisable N determined by waterlogged incubation of laboratory PPS-amended soil samples increased with increasing application rate of Wet PPS. Anaerobically minemlisable N from field-moist soil was well correlated with net N mineralised during 30 weeks of aerobic leached incubation (R^2 =0.90 sandy soil; R^2=0.93 clay soil). In the clay soil, the amount of mineral N produced from all the laboratory incubations was significantly correlated with field-measured nitrate-N in the soil profile (0-1.5 m depth) after 9 months of weed-free fallow following PPS application. In contrast, only anaerobic mineralisable N was significantly correlated with field nitrate-N in the sandy soil. Anaerobic incubation would, therefore, be suitable as a rapid practical test to estimate potentially mineralisable N following applications of different PPS materials in the field.

Field evaluation of a plant activator, captan, chlorothalonil, copper hydroxide, iprodione, mancozeb and strobilurins for the control of citrus brown spot of mandarin

Brown spot (caused by Alternaria alternata) is a major disease of citrus in subtropical areas of Australia. A number of chemicals, the strobilurins azoxystrobin, trifloxystrobin, pyraclostrobin and methoxycrylate, a plant activator (acibenzolar), copper hydroxide, mancozeb, captan, iprodione and chlorothalonil/pyrimthanil were tested in the field for its control. Over three seasons, trees in a commercial orchard received 16, 14 and 7 fungicide sprays, respectively, commencing at flowering in the first season, and petal fall in the later seasons. In all experiments, the strobilurins used alone, or incorporated with copper and mancozeb, were as effective as, or better than the industry standard of copper and mancozeb alone. The only exception was trifloxystrobin, which when used alone was less effective than the industry standard. Acibenzolar used alone was ineffective. Applying a mixture of azoxystrobin and acibenzolar was found to reduce the incidence of brown spot compared with applying azoxystrobin alone but, in either case, disease levels were not found to be significantly different to the industry standard. Captan, iprodione and chlorothalonil/pyrimthanil were as effective as the industry standard. The incidence and severity of rind damage were significantly lowest in the azoxystrobin, methoxycrylate, iprodione and chlorothalonil/pyrimthanil treatments. Medium and high rates of trifloxystrobin (0.07 g/L, 0 .15 g/L) and pyraclostrobin (0.8 g/L, 1.2 g/L) applied alone were the only treatments found to be IPM-incompatible as shown by the elevated level of scale infection on fruit.

Field evaluation of micropropagated and conventionally propagated ginger in subtropical Queensland

The growth and performance of micropropagated ginger (Zingiber officinale Roscoe) was compared with 'seed'-derived plants in field trials conducted in south-eastern Queensland. In the first generation ex vitro, micropropagated plants had significantly (P<0.01) reduced rhizome yield with smaller knobs and more roots. Micropropagated plants had a greater (P<0.01) shoot: root (rhizome) ratio compared with seed-derived plants. Shoots from micropropagated plants were also significantly (P<0.01) smaller with a greater number of shoots per plant. The unusual shoot morphology of the micropropagated plants did not appear to be related to the presence of benzylaminopurine, a plant growth hormone added to the multiplication medium, as plants subcultured for 3 cycles on a hormone-free medium also exhibited similar characteristics. Seed collected from the micropropagated plants and seed-derived plants was harvested and, despite the micropropagated seed being significantly (P<0.01) smaller, by the second generation ex vitro there were no significant differences between the treatments. Factors that can improve rhizome size, while reducing production costs, need to be identified before micropropagated plants can be recommended for routine use in the ginger industry as a source of disease and pest-free planting material.

Potential Human Exposure to Australian Bat Lyssavirus, Queensland, 1996-1999

Two human deaths caused by Australian bat lyssavirus (ABL) infection have been reported since 1996. Information was obtained from 205 persons (mostly adults from south Brisbane and the South Coast of Queensland), who reported potential ABL exposure to the Brisbane Southside Public Health Unit from November 1,1996, to January 31, 1999. Volunteer animal handlers accounted for 39% of potential exposures, their family members for 12%, professional animal handlers for 14%, community members who intentionally handled bats for 31%, and community members with contacts initiated by bats for 4%. The prevalence of Lyssavirus detected by fluorescent antibody test in 366 sick, injured, or orphaned bats from the area was 6%. Sequelae of exposure, including the requirement for expensive postexposure prophylaxis, may be reduced by educating bat handlers and the public of the risks involved in handling Australian bats.

Reclassification of [Pasteurella] trehalosi as Bibersteinia trehalosi gen. nov., comb. nov.

[Pasteurella] trehalosi is an important pathogen of sheep, being primarily associated with serious systemic infections in lambs but also having an association with pneumonia. The aim of the present investigation was to characterize a broad collection of strains tentatively identified as [P.] trehalosi in order to reclassify and rename this taxon to support improvements in our understanding of the pathogenesis and epidemiology of this important organism. The type strain for [P.] trehalosi, strain NCTC 10370T, was included along with 42 field isolates from sheep (21), cattle (14), goats (1), roe deer (3) and unknown sources (3). An extended phenotypic characterization was performed on all 43 strains. Amplified fragment length polymorphism (AFLP) was also performed on the isolates. Two of the field isolates were subjected to 16S rRNA gene sequencing. These sequences, along with five existing sequences for [P.] trehalosi strains and 12 sequences for other taxa in the family Pasteurellaceae, were subjected to a phylogenetic analysis. All the isolates and the reference strains were identified as [P.] trehalosi. A total of 17 out of 22 ovine isolates produced acid from all glycosides, while only four out of 14 bovine isolates produced acid from all glycosides. All 22 ovine isolates were haemolytic and CAMP-positive, while no other isolate was haemolytic and only two bovine isolates were CAMP-positive. Nineteen AFLP types were found within the [P.] trehalosi isolates. All [P.] trehalosi isolates shared at least 70% similarity in AFLP patterns. The largest AFLP type included the type strain and 7 ovine field isolates. Phylogenetic analysis indicated that the seven strains studied (two field isolates and the five serovar reference strains) are closely related, with 98.6% or higher 16S rRNA gene sequence similarity. As both genotypic and phenotypic testing support the separate and distinct nature of these organisms, we propose the transfer of [P.] trehalosi to a new genus, Bibersteinia, as Bibersteinia trehalosi comb. nov. The type strain is NCTC 10370T (=ATCC 29703T). Bibersteinia trehalosi can be distinguished from the existing genera of the family by the observation of only nine characteristics; catalase, porphyrin, urease, indole, phosphatase, acid from dulcitol, (+)-D-galactose, (+)-D mannose and (+)-D-trehalose.

Pasteurella multocida detection by 5' Taq nuclease assay: a new tool for use in diagnosing fowl cholera

A 5′ Taq nuclease assay utilising minor groove binder technology and targeting the 16S rRNA gene was designed to detect Pasteurella multocida (the causative agent of fowl cholera) in swabs collected from poultry. The assay was first evaluated using pure cultures. The assay correctly identified four P. multocida taxonomic type strains, 18 P. multocida serovar reference strains and 40 Australian field isolates (17 from poultry, 11 from pigs and 12 from cattle). Representatives of nine other Pasteurella species, 26 other bacterial species (18 being members of the family Pasteurellaceae) and four poultry virus isolates did not react in the assay. The assay detected a minimum of approximately 10 cfu of P. multocida per reaction. Of 79 poultry swabs submitted to the laboratory for routine bacteriological culture, 17 were positive in the 5′ Taq nuclease assay, but only 10 were positive by culture. The other 62 swabs were negative for P. multocida by both 5′ Taq nuclease assay and culture. The assay is suitable for use in diagnosing fowl cholera, is more rapid than bacteriological culture, and may also have application in diagnosing P. multocida infections in cattle and pigs.

Minimising mortality in field and pot experiments using Heteronyx piceus Blanchard (Coleoptera: Scarabaeidae) eggs and larvae

The establishment of experimental populations of scarab larvae using eggs and early instar larvae has proven to be difficult for many researchers. Despite this, little work has been published examining ways to optimise establishment under artificial conditions. In this experiment, we examined the effect of shade and irrigation on the establishment of Heteronyx piceus Blanchard larvae introduced into pots as eggs and first-, second- and third-instar larvae to optimise artificial infestation techniques. The most important factor affecting larval establishment was the life stage introduced. Establishment of eggs and first instars was very low, with only 21% of eggs and 11% of first-instar larvae establishing. In contrast, 82% of second-instar larvae and 84% of third-instar larvae established successfully. The addition of shade marginally improved overall survival from 45% in the unshaded pots to 53% in the shaded pots. However, most of this increase was in the eggs and first instars. Irrigation did not improve survival. These results suggest that when introducing scarab larvae to field or pot experiments, second- or thirdinstar larvae should be used to maximise establishment. The provision of shade and supplementary irrigation is optional.

Review of Nezara viridula (L.) management strategies and potential for IPM in field crops with emphasis on Australia

Nezara viridula (L.) is a cosmopolitan, polyphagous heteropteran that causes economic damage to many crop species. At present, control of N. viridula in Australia and other countries relies heavily upon insecticides, most of which are disruptive to beneficial insects, constituting a constraint on integrated pest management (IPM). Much research has been conducted into non-chemical control methods for N. viridula. This paper reviews the potential for and limitations of sterile insect technique, classical, inundative and conservation biological control, and trap cropping. None of these techniques appear to be adequate for control of N. viridula when used alone but there is scope for these non-chemical approaches to be adopted for use in integrated management of this pest. A proposal is given for one such integrated approach for future development. It includes biopesticides, trap crops and carefully targeted habitat manipulation to enhance arthropod natural enemies as well as area-wide management and grower education.

Limitations of soil microbial biomass carbon as an indicator of soil pollution in the field

The size of the soil microbial biomass carbon (SMBC) has been proposed as a sensitive indicator for measuring the adverse effects of contaminants on the soil microbial community. In this study of Australian agricultural systems, we demonstrated that field variability of SMBC measured using the fumigation-extraction procedure limited its use as a robust ecotoxicological endpoint. The SMBC varied up to 4-fold across control samples collected from a single field site, due to small-scale spatial heterogeneity in the soil physicochemical environment. Power analysis revealed that large numbers of replicates (3-93) were required to identify 20% or 50% decreases in the size of the SMBC of contaminated soil samples relative to their uncontaminated control samples at the 0.05% level of statistical significance. We question the value of the routine measurement of SMBC as an ecotoxicological endpoint at the field scale, and suggest more robust and predictive microbiological indicators.

Detection of Chlamydiaceae DNA in veterinary specimens using a family-specific PCR

Aims: The aim of this work was to develop a rapid molecular test for the detection of the Chlamydiaceae family, irrespective of the species or animal host. Methods and Results: The method described herein is a polymerase chain reaction targeting the 16S rRNA gene of the Chlamydiaceae family, and the results demonstrate that the test reacts with five reference Chlamydiaceae but none of the 19 other bacterial species or five uninfected animal tissues tested. The results also indicate the enhanced sensitivity of this test when compared with conventional culture or serology techniques. This is demonstrated through parallel testing of six real clinical veterinary cases and confirmatory DNA sequence analysis. Conclusions, Significance and Impact of the Study: This test can be used by veterinary diagnostic laboratories for rapid detection of Chlamydiaceae in veterinary specimens, with no restriction of chlamydial species or animal host. The test does not differentiate chlamydial species, and if required, speciation must be carried out retrospectively using alternate methods. However, for the purpose of prescribing therapy for chlamydiosis, this test would be an invaluable laboratory tool.

Baseline responses of Alphitobius diaperinus (Coleoptera : Tenebrionidae) to cyfluthrin and detection of strong resistance in field populations in eastern Australia

Resistance to cyfluthrin in broiler farm populations of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in eastern Australia was suspected to have contributed to recent control failures. In 2000-2001, beetles from 11 broiler farms were tested for resistance by comparing them to an insecticide-susceptible reference population by using topical application. Resistance was detected in almost all beetle populations (up to 22 times the susceptible at the LC50), especially in southeastern Queensland where more cyfluthrin applications had been made. Two from outside southeastern Queensland were found to be susceptible. Dose-mortality data generated from the reference population over a range of cyflutbrin concentrations showed that 0.0007% cyfluthrin at a LC99.9 level could be used as a convenient dose to discriminate between susceptible and resistant populations. Using this discriminating concentration, from 2001 to 2005, the susceptibilities of 18 field populations were determined. Of these, 11 did not exhibit complete mortality at the discriminating concentration (mortality range 2.8-97.7%), and in general, cyfluthrin resistance was directly related to the numbers of cyfluthrin applications. As in the full study, populations outside of southeastern Queensland were found to have lower levels of resistance or were susceptible. One population from an intensively farmed broiler area in southeastern Queensland exhibited low mortality despite having no known exposure to cyfluthrin. Comparisons of LC50 values of three broiler populations and a susceptible population, collected in 2000 and 2001 and recollected in 2004 and 2005 indicated that values from the three broiler populations had increased over this time for all populations. The continued use of cyfluthrin for control of A. diaperinus in eastern Australia is currently under consideration.

Abaca bunchy top virus, a new member of the genus Babuvirus (family Nanoviridae)

Two isolates of a novel babuvirus causing "bunchy top" symptoms were characterised, one from abaca (Musa textilis) from the Philippines and one from banana (Musa sp.) from Sarawak (Malaysia). The name abacá bunchy top virus (ABTV) is proposed. Both isolates have a genome of six circular DNA components, each ca. 1.0-1.1 kb, analogous to those of isolates of Banana bunchy top virus (BBTV). However, unlike BBTV, both ABTV isolates lack an internal ORF in DNA-R, and the ORF in DNA-U3 found in some BBTV isolates is also absent. In all phylogenetic analyses of nanovirid isolates, ABTV and BBTV fall in the same clade, but on separate branches. However, ABTV and BBTV isolates shared only 79-81% amino acid sequence identity for the putative coat protein and 54-76% overall nucleotide sequence identity across all components. Stem-loop and major common regions were present in ABTV, but there was less than 60% identity with the major common region of BBTV. ABTV and BBTV were also shown to be serologically distinct, with only two out of ten BBTV-specific monoclonal antibodies reacting with ABTV. The two ABTV isolates may represent distinct strains of the species as they are less closely related to each other than are isolates of the two geographic subgroups (Asian and South Pacific) of BBTV.

Field assessment of the resistance of timber products to termite (Isoptera) damage: Results can change with the amount of bait wood

Whether or not termites initiate damage to timber via the end grain may determine the need for spot-treating the exposed untreated cut ends of envelope-treated softwood framing material. Australian Coptotermes acinaciformis (Froggatt) were field-tested for their ability to initiate feeding via the end grain of timber (35 × 90 mm) treated with a repellent Tanalith® T envelope. Specimens of commercial radiata pine Pinus radiata D.Don framing timber (untreated) and slash pine Pinus elliottii Englem. (untreated and envelope-treated) were partially clad in fine stainless steel mesh. Clad and unclad specimens were exposed to C. acinaciformis near Townsville, North Queensland, Australia, for four months. Results showed that this species of termite can indeed damage timber via the end grain, including exposed untreated cut ends of envelope-treated material as demonstrated earlier for different populations of C. acinaciformis. Differences between the test conditions in field trials carried out at different times (where C. acinaciformis either did or did not damage timber via the end grain) are discussed. Clearly, outcomes from field studies with preservative-treated materials are dependent upon experimental conditions. Notably, the amount of bait wood (highly termite-susceptible timber substrate) offered in a given method can strongly influence the termite response. Further investigation is required to standardise this aspect of conditions in protocols for the assessment of wood preservatives.

Genetic control of propagation traits in a single Corymbia torelliana x Corymbia variegata family

Genetic control of vegetative propagation traits was described for a second-generation, outbred, intersectional hybrid family (N = 208) derived from two species, Corymbia torelliana (F. Muell.) K.D. Hill & L.A.S. Johnson and Corymbia variegata (F. Muell.) K.D. Hill & L.A.S. Johnson, which contrast for propagation characteristics and in their capacity to develop lignotubers. Large phenotypic variances were evident for rooting and most other propagation traits, with significant proportions attributable to differences between clones (broad-sense heritabilities 0.2-0.5). Bare root assessment of rooting rate and root quality parameters tended to have the highest heritabilities, whereas rooting percentage based on root emergence from pots and shoot production were intermediate. Root biomass and root initiation had the lowest heritabilities. Strong favourable genetic correlations were found between rooting percentage and root quality traits such as root biomass, volume, and length. Lignotuber development on a seedling was associated with low rooting and a tendency to poor root quality in cuttings and was in accord with the persistence of species parent types due to gametic phase disequilibrium. On average, nodal cuttings rooted more frequently and with higher quality root systems, but significant cutting type x genotype interaction indicated that for some clones, higher rooting rates were obtained from tips. Low germination, survival of seedlings, and rooting rates suggested strong hybrid breakdown in this family.