Investigation and application of methods for enumerating heterotrophs and Escherichia coli in the air within piggery sheds

Aims: To investigate methods for the recovery of airborne bacteria within pig sheds and to then use the appropriate methods to determine the levels of heterotrophs and Escherichia coli in the air within sheds. Methods and Results: AGI-30 impingers and a six-stage Andersen multi-stage sampler (AMS) were used for the collection of aerosols. Betaine and catalase were added to impinger collection fluid and the agar plates used in the AMS. Suitable media for enumerating E. coli with the Andersen sampler were also evaluated. The addition of betaine and catalase gave no marked increase in the recovery of heterotrophs or E. coli. No marked differences were found in the media used for enumeration of E. coli. The levels of heterotrophs and E. coli in three piggeries, during normal pig activities, were 2Æ2 · 105 and 21 CFU m)3 respectively. Conclusions: The failure of the additives to improve the recovery of either heterotrophs or E. coli suggests that these organisms are not stressed in the piggery environment. The levels of heterotrophs in the air inside the three Queensland piggeries investigated are consistent with those previously reported in other studies. Flushing with ponded effluent had no marked or consistent effect on the heterotroph or E. coli levels. Significance and Impact of the Study: Our work suggests that levels of airborne heterotrophs and E. coli inside pig sheds have no strong link with effluent flushing. It would seem unlikely that any single management activity within a pig shed has a dominant influence on levels of airborne heterotrophs and E. coli

The process for implementation of a Quality Management System within a multi-functional cereal laboratory

The intent of this study was to design, document and implement a Quality Management System (QMS) into a laboratory that incorporated both research and development (R&D) and routine analytical activities. In addition, it was necessary for the QMS to be easily and efficiently maintained to: (a) provide documented evidence that would validate the system's compliance with a certifiable standard, (b) fit the purpose of the laboratory, (c) accommodate prevailing government policies and standards, and (d) promote positive outcomes for the laboratory through documentation and verification of the procedures and methodologies implemented. Initially, a matrix was developed that documented the standards' requirements and the necessary steps to be made to meet those requirements. The matrix provided a check mechanism on the progression of the system's development. In addition, it was later utilised in the Quality Manual as a reference tool for the location of full procedures documented elsewhere in the system. The necessary documentation to build and monitor the system consisted of a series of manuals along with forms that provided auditable evidence of the workings of the QMS. Quality Management (QM), in one form or another, has been in existence since the early 1900's. However, the question still remains: is it a good thing or just a bugbear? Many of the older style systems failed because they were designed by non-users, fiercely regulatory, restrictive and generally deemed to be an imposition. It is now considered important to foster a sense of ownership of the system by the people who use the system. The system's design must be tailored to best fit the purpose of the operations of the facility if maximum benefits to the organisation are to be gained.

Effect of fragmentation, habitat loss and within-patch habitat characteristics on ant assemblages in semi-arid woodlands of eastern Australia.

The reliability of ants as bioindicators of ecosystem condition is dependent on the consistency of their response to localised habitat characteristics, which may be modified by larger-scale effects of habitat fragmentation and loss. We assessed the relative contribution of habitat fragmentation, habitat loss and within-patch habitat characteristics in determining ant assemblages in semi-arid woodland in Queensland, Australia. Species and functional group abundance were recorded using pitfall traps across 20 woodland patches in landscapes that exhibited a range of fragmentation states. Of fragmentation measures, changes in patch area and patch edge contrast exerted the greatest influence on species assemblages, after accounting for differences in habitat loss. However, 35% of fragmentation effects on species were confounded by the effects of habitat characteristics and habitat loss. Within-patch habitat characteristics explained more than twice the amount of species variation attributable to fragmentation and four times the variation explained by habitat loss. The study indicates that within-patch habitat characteristics are the predominant drivers of ant composition. We suggest that caution should be exercised in interpreting the independent effects of habitat fragmentation and loss on ant assemblages without jointly considering localised habitat attributes and associated joint effects.

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.

An assessment of the genetic relationship between sweet and grain sorghums, within Sorghum bicolor ssp. bicolor (L.) Moench, using AFLP markers

Compared to grain sorghums, sweet sorghums typically have lower grain yield and thick, tall stalks which accumulate high levels of sugar (sucrose, fructose and glucose). Unlike commercial grain sorghum (S. bicolor ssp. bicolor) cultivars, which are usually F1 hybrids, commercial sweet sorghums were selected as wild accessions or have undergone limited plant breeding. Although all sweet sorghums are classified within S. bicolor ssp. bicolor, their genetic relationship with grain sorghums is yet to be investigated. Ninety-five genotypes, including 31 sweet sorghums and 64 grain sorghums, representing all five races within the subspecies bicolor, were screened with 277 polymorphic amplified fragment length polymorphism (AFLP) markers. Cluster analysis separated older sweet sorghum accessions (collected in mid 1800s) from those developed and released during the early to mid 1900s. These groups were emphasised in a principle component analysis of the results such that sweet sorghum lines were largely distinguished from the others, particularly by a group of markers located on sorghum chromosomes SBI-08 and SBI-10. Other studies have shown that QTL and ESTs for sugar-related traits, as well as for height and anthesis, map to SBI-10. Although the clusters obtained did not group clearly on the basis of racial classification, the sweet sorghum lines often cluster with grain sorghums of similar racial origin thus suggesting that sweet sorghum is of polyphyletic origin within S. bicolor ssp. bicolor.

Congruence in QTL for adventitious rooting in Pinus elliottii x Pinus caribaea hybrids resolves between and within-species effects

Targeting between-species effects for improvement in synthetic hybrid populations derived from outcrossing parental tree species may be one way to increase the efficacy and predictability of hybrid breeding. We present a comparative analysis of the quantitative trait loci (QTL) which resolved between from within-species effects for adventitious rooting in two populations of hybrids between Pinus elliottii and P. caribaea, an outbred F1 (n=287) and an inbred-like F2 family (n=357). Most small to moderate effect QTL (each explaining 2-5% of phenotypic variation, PV) were congruent (3 out of 4 QTL in each family) and therefore considered within-species effects as they segregated in both families. A single large effect QTL (40% PV) was detected uniquely in the F2 family and assumed to be due to a between-species effect, resulting from a genetic locus with contrasting alleles in each parental species. Oligogenic as opposed to polygenic architecture was supported in both families (60% and 20% PV explained by 4 QTL in the F 2 and F1 respectively). The importance of adventitious rooting for adaptation to survive water-logged environments was thought in part to explain oligogenic architecture of what is believed to be a complex trait controlled by many hundreds of genes.

Genetic variation within two sympatric spotted gum eucalypts exceeds between taxa variation

Population substructure and hybridization, among other factors, have the potential to cause erroneous associations in linkage disequilibrium (LD) mapping. Two closely related spotted gum eucalypts, Corymbia variegata and C. henryi (Myrtaceae) occur in sympatry in the east coast of Australia and potentially interbreed. They are morphologically similar but are distinguished as separate species based on capsule and foliage size. To determine whether they hybridize in nature and its implications for LD mapping, we investigated the level of molecular divergence between the two species at two sympatric locations separated by 300 kilometres. Very few individuals of intermediate morphology were identified, despite the two species occurring only metres apart. Analysis of genetic structure using 12 microsatellite loci showed that genetic differentiation between populations of the same species at different locations (FST = 0.07 for both species; p = 0.0001) was significantly higher than that observed between species at each location (mean FST = 0.02 and 0.04 for Cherry tree and Bunyaville respectively; p = 0.0001; all Mann-Whitney U-test p ≤ 0.01). No species-specific alleles or significant allele frequency differences were detected within a site, suggesting recurrent local gene flow between the two species. The lack of significant allele frequency differences implies no population stratification along taxonomic lines. This suggested that there is little concern for cryptic hybridization when sampling from sites of sympatry for LD mapping.

DNA amplification fingerprinting analysis of genetic variation within Fusarium oxysporum f.sp zingiberi

Genetic variation among 29 isolates of Fusarium oxysporum f.sp. zingiberi (Foz) collected from diseased ginger rhizome in production regions throughout Queensland was analysed using DNA amplification fingerprinting (DAF). Eight isolates of other Fusarium species and/or formae speciales were included for comparative analysis. Within the Foz isolates, three haplotypes were identified based on 17 polymorphic bands generated with five primers. Two groups showed very little genetic variation (98.6% similarity), whereas the third single isolate was quite distinct in terms of its molecular profile (77.2% similarity). Genetic similarity among the Fusarium solani, F. oxysporum f.sp. lycopersici and F. oxysporum f.sp. cubense races 1, 3 and 4 isolates compared well with the published literature.

Molecular and serological detection of A. centrale- and A. marginale-infected cattle grazing within an endemic area

A reverse line blot hybridization (RLB) one-stage nested PCR (nPCR) for Anaplasma centrale and a nested PCR for Anaplasma marginale were used to detect infected cattle grazing within an endemic region in Israel. A novel set of PCR primers and oligonucleotide probes based on a 16S ribosomal RNA gene was designed for RLB detection of both Anaplasma species, and the performance of the molecular assays compared. The immunofluorescent antibody test (IFA) was used to detect antibodies to both Anaplasma species, whereas, a highly sensitive and specific competitive enzyme-linked immunosorbent assay (cELISA) was used to detect antibodies in A. centrale-vaccinated cattle. The RLB and the nested PCR procedures showed bacteremia with sensitivity of 50 infected erythrocytes per milliliter. Up to 93% of the A. centrale vaccinates carried specific antibodies that were detected by cELISA, and up to 71% of the vaccinated cattle were found to be naturally infected with A. marginale according to the PCR and the RLB assays. Nevertheless, no severe outbreaks of A. marginale infection occurred among vaccinated herds in this endemic region. It appears that both, molecular tools and serology are useful for evaluation of the vaccine efficacy. In the light of wide natural field infection with A. marginale, strong recommendations to continue the A. centrale vaccination program regime will continue until a new generation of non-blood-based vaccine will be developed.

The assessment of TGGE for the detection of interspecific and intergeneric DNA-marker polymorphism within Solanaceae

RFLP markers are currently the most appropriate marker system for the identification of uncharacterised polymorphism at the interspecific and intergeneric level. Given the benefits of a PCR-based marker system and the availability of sequence information for many Solanaceous cDNA clones, it is now possible to target conserved fragments, for primer development, that flank sequences possessing interspecific polymorphism. The potential outcome is the development of a suite of markers that amplify widely in Solanaceae. Temperature gradient gel electrophoresis (TGGE) is a relatively inexpensive gel-based system that is suitable for the detection of most single-base changes. TGGE can be used to screen for both known and unknown polymorphisms, and has been assessed here, for the development of PCR-based markers that are useful for the detection of interspecific variation within Solanaceae. Fifteen markers are presented where differences between Lycopersicon esculentum and L. pennellii have been detected by TGGE. The markers were assessed on a wider selection of plant species and found to be potentially useful for the identification of interspecific and intergeneric polymorphism in Solanaceous plants.

Optimising between and within family selection for harvest weight in prawns with restricted harvest size.

Manual grading of prawns restricts the number that can be harvested. A restricted harvest size places a limit on the opposing within family and between family sources of selection pressure. A simulation study with inbreeding constrained at 0.5% per generation, a harvest size of 2000, heritability of 0.3, common family environmental effect of 0.1, indicates that maximum response to selection could be achieved with as few as 40 families. Increasing the number of families above 80 may reduce total selection response. It is important to be aware that increasing the number of families may not always yield a greater genetic response.

Evidence for extensive population structure in the white-spotted eagle ray within the Indo-Pacific inferred from mitochondrial gene sequences

The white-spotted eagle ray Aetobatus narinari is a species complex that occurs circumglobally throughout warm-temperate waters. Aetobatus narinari is semi-pelagic and large (up to 300 cm disc width), suggesting high dispersal capabilities and gene flow on a wide spatial scale. Sequence data from two mitochondrial genes, cytochrome b (cytb) and NADH dehydrogenase subunit 4 (ND4), were used to determine the genetic variability within and among 18 sampling locations in the central Indo-Pacific biogeographical region. Populations in the Indo-Pacific were highly genetically structured with c. 70% of the total genetic variation found among three geographical regions (East China Sea, Southeast Asia and Australia). FST was 0.64 for cytb and 0.53 for ND4, with φST values being even larger, that is, 0.78 for cytb and 0.65 for ND4. This high-level genetic partitioning provides strong evidence against extensive gene flow in A. narinari. The degree of genetic population structuring in the Indo-Pacific was similar to that found on a global scale. Global FST was 0.63 for cytb and 0.57 for ND4, and global φST values were 0.94 for cytb and 0.82 for ND4. This suggests that the A. narinari complex may be more speciose than the two or three species proposed to date. Further sampling and genetic analyses are likely to uncover the ‘evolutionarily significant’ and ‘management’ units that are critical to determine the susceptibilities of individual populations to regional fishing pressures and to provide advice on management options. Network analyses showed a close genetic relationship between haplotypes from the central Indo-Pacific and South Africa, providing support for a proposed dispersal pathway from the possible centre of origin of the A. narinari species complex in the Indo-Pacific into the Atlantic Ocean.

Role of Native Fish in Integrated Aquatic Weed and Water Quality Management within the Burdekin Irrigation Area

This report summarises work conducted by the QDPI, in partnership with the South Burdekin Water Board (SBWB) and the Burdekin Shire Council (BSC) between 2001 and 2003. The broad aim of the research was to assess the potential of native fish as biocontrol agents for noxious weeds, as part of an integrated program for managing water quality in the Burdekin Irrigation Area. A series of trials were conducted at, or using water derived from, the Sandy Creek Diversion near Groper Creek (lower Burdekin delta). Trials demonstrated that aquatic weeds play a positive role in trapping transient nutrients, until such time that weed growth becomes self-shading and weed dieback occurs, which releases stored nutrients and adversely affects water quality. Transient nutrient levels (av. TN<0.5mg/L; av. TP<0.1mg/L) found in the irrigation channel during the course of this research were substantially lower than expected, especially considering the intensive agriculture and sewage effluent discharge upstream from the study site. This confirms the need to consider the control of weeds rather than complete weed extermination when formulating management plans. However, even when low nutrient levels are available, there is competitive exploitation of habitat variables in the irrigation area leading to succession and eventual domination by certain weed species. During these trials, we have seen filamentous algae, phytoplankton, hyacinth and curled pondweed each hold competitive advantage at certain points. However without intervention, floating weeds, especially hyacinth, ultimately predominate in the Burdekin delta due to their fast propagation rate and their ability to out-shade submerged plants. We have highlighted the complexity of interactions in these highly disturbed ecosystems in that even if the more prevalent noxious weeds are contained, other weed species will exploit the vacant niche. This complexity places stringent requirements on the type of native fish that can be used as biocontrol agents. Of the seven fish species identified with herbivorous trophic niches, most target plankton or algae and do not have the physical capacity to directly eat the larger macrophytes of the delta. We do find however that following mechanical weed harvesting, inoculative releases of fish can slow the rate of hyacinth recolonisation. This occurs by mechanisms in addition to direct weed consumption, such as disturbing growth surfaces by grazing on attached biofilms. Predation by birds and water rats presents another impediment to the efficacy of large-scale releases of fish. However, alternative uses of fish in water quality management in the Burdekin irrigation area are discussed.

Species within Mycosphaerellaceae and Teratosphaeriaceae from eucalypts in eastern Australia

Twenty eight species within Mycosphaerellaceae and Teratosphaeriaceae (includes Mycosphaerella, Teratosphaeria, Pseudocercospora and Sonderhenia) are reported from Eucalyptus and Corymbia in New South Wales and Queensland, Australia, based on field surveys and examination of herbarium specimens and published reports. Teratosphaeria cryptica was the most commonly recorded species, with the widest host range and distribution, followed by Mycosphaerella marksii. Six new species are described: T. keanei, T. coolabuniensis, T. crispata, M. medusae, M. nootherensis and T. praelongispora. New or interesting records for known species are reported, including new records for Australia (T. pluritubularis and T. verrucosiafricana) and new records for Queensland (T. excentrica, T. multiseptata, T. nubilosa, T. suberosa and Ps. pseudoeucalyptorum).

Background to the approved beneficial Cynodon spp. vegetative cultivars within Australia

The growth of the Australian turfgrass industry has significantly expanded over recent decades. One reason for this occurring has been with development of better suited or higher quality turfgrass cultivars for Australia’s harsh climatic conditions. In recent years drought has widely affected the turfgrass industry and as such, greater drought tolerant C4 grasses such as Cynodon spp. have been used. In 2008, as part of the 24th Australian Turfgrass Conference Proceedings, Peter McMaugh wrote an extensive article on the couch grass breeding history in Australia. This paper contains an extension to his work detailing the current (1950s to 2010) Cynodon species found in Australia. Detailed information has been sourced in relation to the origin and development of the grasses which are suitable for turfgrass use. Such detail provides an interesting picture of the source of proliferation of newer cultivars and how the Australian industry has evolved with the introduction of overseas and Australian selected cultivars. The information adds to the preceding work, including morphological and agronomic attributes and how closely each selection or cultivar is related. The cultivars discussed in this article (listed alphabetically) are derived from one of the four classifications identified by the breeder/author, being (i) Cynodon sp. (although the cultivar contained within the taxa fits best being classified as a Cynodon hybrid), (ii) Cynodon dactylon x C. transvaalensis (Cynodon hybrid), (iii) Cynodon dactylon (green couch) and (iv) Cynodon dactylon x C. magenissii.