Isolation of Pseudomonas pseudomallei from clay layers at defined depths

Twelve strains of Pseudomonas pseudomallei were isolated from the soil and water of a sheep paddock over a two-year period. The organism was recovered from the clay layer of the soil profile as well as from water that seeps into this layer during the "wet" season. Five isolates were obtained before the commencement of the "wet" season; environmental factors appear to play an important role in the survival of Ps. pseudomallei during the "dry" season. Lower isolation rates were recorded than those indicated by workers in southeast Asia and Iran.

Influence of environmental and instrumental variables on the non-invasive prediction of Brix in pineapple using near infrared spectroscopy

The Brix content of pineapple fruit can be non-invasively predicted from the second derivative of near infrared reflectance spectra. Correlations obtained using a NIRSystems 6500 spectrophotometer through multiple linear regression and modified partial least squares analyses using a post-dispersive configuration were comparable with that from a pre-dispersive configuration in terms of accuracy (e.g. coefficient of determination, R2, 0.73; standard error of cross validation, SECV, 1.01°Brix). The effective depth of sample assessed was slightly greater using the post-dispersive technique (about 20 mm for pineapple fruit), as expected in relation to the higher incident light intensity, relative to the pre-dispersive configuration. The effect of such environmental variables as temperature, humidity and external light, and instrumental variables such as the number of scans averaged to form a spectrum, were considered with respect to the accuracy and precision of the measurement of absorbance at 876 nm, as a key term in the calibration for Brix, and predicted Brix. The application of post-dispersive near infrared technology to in-line assessment of intact fruit in a packing shed environment is discussed.

Cytochrome P450 aromatase in grey mullet: cDNA and promoter isolation; brain, pituitary and ovarian expression during puberty.

In a study towards elucidating the role of aromatases during puberty in female grey mullet, the cDNAs of the brain (muCyp19b) and ovarian (muCyp19a) aromatase were isolated by RT-PCR and their relative expression levels were determined by quantitative real-time RT-PCR. The muCyp19a ORF of 1515 bp encoded 505 predicted amino acid residues, while that of muCyp19b was 1485 bp and encoded 495 predicted amino acid residues. The expression level of muCyp19b significantly increased in the brain as puberty advanced; however, its expression level in the pituitary increased only slightly with pubertal development. In the ovary, the muCyp19a expression level markedly increased as puberty progressed. The promoter regions of the two genes were also isolated and their functionality evaluated in vitro using luciferase as the reporter gene. The muCyp19a promoter sequence (650 bp) contained a consensus TATA box and putative transcription factor binding sites, including two half EREs, an SF-1, an AhR/Arnt, a PR and two GATA-3s. The muCyp19b promoter sequence (2500 bp) showed consensus TATA and CCAAT boxes and putative transcription binding sites, namely: a PR, an ERE, a half ERE, a SP-1, two GATA-binding factor, one half GATA-1, two C/EBPs, a GRE, a NFkappaB, three STATs, a PPAR/RXR, an Ahr/Arnt and a CRE. Basal activity of serially deleted promoter constructs transiently transfected into COS-7, [alpha]T3 and TE671 cells demonstrated the enhancing and silencing roles of the putative transcription factor binding sites. Quinpirole, a dopamine agonist, significantly reduced the promoter activity of muCyp19b in TE671. The results suggest tissue-specific regulation of the muCyp19 genes and a putative alternative promoter for muCyp19b.

Assessing for genetic and environmental effects on ruminant feed quality in barley ( Hordeum vulgare )

Grain samples from a combined intermediate and advanced stage barley breeding trial series, grown at two sites in two consecutive years were assessed for detailed grain quality and ruminant feed quality. The results indicated that there were significant genetic and environmental effects for “feed” traits as measured using grain hardness, acid detergent fibre (ADF), starch and in-sacco dry matter digestibility (ISDMD) assays. In addition, there was strong genotypic discrimination for the regressed feed performance traits, namely Net Energy (NE) and Average Daily Gain (ADG). There was considerable variation in genetic correlations for all traits based on variance from the cultivars used, sites or laboratory processing effects. There was a high level of heritability ranging from 89% to 88% for retention, 60% to 80% for protein and 56% to 68% for ADF. However, there were only low to moderate levels of heritability for the feed traits, with starch 30–39%, ISDMD 55–63%, ADF 56–68%, particle size 47–73%, 31–48% NE and ADG 44–51%. These results suggest that there were real differences in the feed performance of barleys and that selection for cattle feed quality is potentially a viable option for breeding programs.

Isolation and expression analysis of multiple isoforms of putative farnesoic acid O-methyltransferase in several crustacean species

Farnesoic acid O-methyltransferase (FaMeT) is the enzyme responsible for the conversion of farnesoic acid (FA) to methyl farnesoate (MF) in the final step of MF synthesis. Multiple isoforms of putative FaMeT were isolated from six crustacean species belonging to the families Portunidae, Penaeidae, Scyllaridae and Parastacidae. The portunid crabs Portunus pelagicus and Scylla serrata code for three forms: short, intermediate and long. Two isoforms (short and long) were isolated from the penaeid prawns Penaeus monodon and Fenneropenaeus merguiensis. Two isoforms were also identified in the scyllarid Thenus orientalis and parastacid Cherax quadricarinatus. Putative FaMeT sequences were also amplified from the genomic DNA of P. pelagicus and compared to the putative FaMeT transcripts expressed. Each putative FaMeT cDNA isoform was represented in the genomic DNA, indicative of a multi-gene family. Various tissues from P. pelagicus were individually screened for putative FaMeT expression using PCR and fragment analysis. Each tissue type expressed all three isoforms of putative FaMeT irrespective of sex or moult stage. Protein domain analysis revealed the presence of a deduced casein kinase II phosphorylation site present only in the long isoform of putative FaMeT.

Measurement of genetic and environmental variation in barley (Hordeum vulgare) grain hardness

In this study, we assessed a broad range of barley breeding lines and commercial varieties by three hardness methods (two particle size methods and one crush resistance method (SKCS—Single-Kernel Characterization System), grown at multiple sites to see if there was variation in barley hardness and if that variation was genetic or environmentally controlled. We also developed near-infrared reflectance (NIR) calibrations for these three hardness methods to ascertain if NIR technology was suitable for rapid screening of breeding lines or specific populations. In addition, we used this data to identify genetic regions that may be associated with hardness. There were significant (p<0.05) genetic effects for the three hardness methods. There were also environmental effects, possibly linked to the effect of protein on hardness, i.e. increasing protein resulted in harder grain. Heritability values were calculated at >85% for all methods. The NIR calibrations, with R2 values of >90%, had Standard Error of Prediction values of 0.90, 72 and 4.0, respectively, for the three hardness methods. These equations were used to predict hardness values of a mapping population which resulted in genetic markers being identified on all chromosomes but chromosomes 2H, 3H, 5H, 6H and 7H had markers with significant LOD scores. The two regions on 5H were on the distal end of both the long and short arms. The region that showed significant LOD score was on the long arm. However, the region on the short arm associated with the hardness (hordoindoline) genes did not have significant LOD scores. The results indicate that barley hardness is influenced by both genotype and environment and that the trait is heritable, which would allow breeders to develop very hard or soft varieties if required. In addition, NIR was shown to be a reliable tool for screening for hardness. While the data set used in this study has a relatively low variation in hardness, the tools developed could be applied to breeding populations that have large variation in barley grain hardness.

Isolation and characterization of novel microsatellite markers and their application for diversity assessment in cultivated groundnut (Arachis hypogaea)

Background: Cultivated peanut or groundnut (Arachis hypogaea L.) is the fourth most important oilseed crop in the world, grown mainly in tropical, subtropical and warm temperate climates. Due to its origin through a single and recent polyploidization event, followed by successive selection during breeding efforts, cultivated groundnut has a limited genetic background. In such species, microsatellite or simple sequence repeat (SSR) markers are very informative and useful for breeding applications. The low level of polymorphism in cultivated germplasm, however, warrants a need of larger number of polymorphic microsatellite markers for cultivated groundnut. Results: A microsatellite- enriched library was constructed from the genotype TMV2. Sequencing of 720 putative SSR-positive clones from a total of 3,072 provided 490 SSRs. 71.2% of these SSRs were perfect type, 13.1% were imperfect and 15.7% were compound. Among these SSRs, the GT/CA repeat motifs were the most common (37.6%) followed by GA/CT repeat motifs (25.9%). The primer pairs could be designed for a total of 170 SSRs and were optimized initially on two genotypes. 104 (61.2%) primer pairs yielded scorable amplicon and 46 (44.2%) primers showed polymorphism among 32 cultivated groundnut genotypes. The polymorphic SSR markers detected 2 to 5 alleles with an average of 2.44 per locus. The polymorphic information content (PIC) value for these markers varied from 0.12 to 0.75 with an average of 0.46. Based on 112 alleles obtained by 46 markers, a phenogram was constructed to understand the relationships among the 32 genotypes. Majority of the genotypes representing subspecies hypogaea were grouped together in one cluster, while the genotypes belonging to subspecies fastigiata were grouped mainly under two clusters. Conclusion. Newly developed set of 104 markers extends the repertoire of SSR markers for cultivated groundnut. These markers showed a good level of PIC value in cultivated germplasm and therefore would be very useful for germplasm analysis, linkage mapping, diversity studies and phylogenetic relationships in cultivated groundnut as well as related Arachis species.

Drivers for uptake of environmental management systems by pastoralists in western Queensland

Pastoralists from 37 beef cattle and sheep properties in western Queensland developed and implemented an environmental management system (EMS) over 18 months. The EMS implemented by them was customised for the pastoral industry as part of a national EMS pilot project, and staff from this project encouraged and assisted pastoralists during this trial. The 31 pastoralists surveyed at the end of the pilot project identified few benefits of EMS implementation, and these were largely associated with environmental management and sustainability. In terms of the reasons for uptake of an EMS, these pastoralists identified drivers similar to those reported in other primary industry sectors. These included improving property and environmental management, financial incentives, a range of market benefits, assistance with red tape issues, access to other training opportunities and assistance and support with the development of their EMS. However, these drivers are weak, and are not motivating pastoralists to adopt an EMS. In contrast, barriers to adoption such as the time involved in developing and implementing EMS are tangible and immediate. Given a lack of effective drivers and that pastoralists are under considerable pressure from ongoing rural adjustment processes, it is not surprising that an EMS is a low priority. It is concluded that widespread uptake and on-going use of an EMS in the pastoral industry will not occur unless pastoralists are required or rewarded for this by markets, governments, financiers, and regional natural resource management bodies.

The environmental fate of diuron under a conventional production regime in a sugarcane farm during the plant cane phase

BACKGROUND: Field studies of diuron and its metabolites 3-(3,4-dichlorophenyl)-1-methylurea (DCPMU), 3,4-dichlorophenylurea (DCPU) and 3,4-dichloroaniline (DCA) were conducted in a farm soil and in stream sediments in coastal Queensland, Australia. RESULTS: During a 38 week period after a 1.6 kg ha^-1 diuron application, 70-100% of detected compounds were within 0-15 cm of the farm soil, and 3-10% reached the 30-45 cm depth. First-order t1/2 degradation averaged 49 ± 0.9 days for the 0-15, 0-30 and 0-45 cm soil depths. Farm runoff was collected in the first 13-50 min of episodes lasting 55-90 min. Average concentrations of diuron, DCPU and DCPMU in runoff were 93, 30 and 83-825 µg L^-1 respectively. Their total loading in all runoff was >0.6% of applied diuron. Diuron and DCPMU concentrations in stream sediments were between 3-22 and 4-31 µg kg^-1 soil respectively. The DCPMU/diuron sediment ratio was >1. CONCLUSION: Retention of diuron and its metabolites in farm topsoil indicated their negligible potential for groundwater contamination. Minimal amounts of diuron and DCMPU escaped in farm runoff. This may entail a significant loading into the wider environment at annual amounts of application. The concentrations and ratio of diuron and DCPMU in stream sediments indicated that they had prolonged residence times and potential for accumulation in sediments. The higher ecotoxicity of DCPMU compared with diuron and the combined presence of both compounds in stream sediments suggest that together they would have a greater impact on sensitive aquatic species than as currently apportioned by assessments that are based upon diuron alone.

Crop and environmental attributes underpinning genotype by environment interaction in synthetic-derived bread wheat evaluated in Mexico and Australia

Synthetic backcrossed-derived bread wheats (SBWs) from CIMMYT were grown in the north-west of Mexico (CIANO) and sites across Australia during 3 seasons. A different set of lines was evaluated each season, as new materials became available from the CIMMYT crop enhancement program. Previously, we have evaluated both the performance of genotypes across environments and the genotype x environment interaction (G x E). The objective of this study was to interpret the G x E for yield in terms of crop attributes measured at individual sites and to identify the potential environmental drivers of this interaction. Groups of SBWs with consistent yield performance were identified, often comprising closely related lines. However, contrasting performance was also relatively common among sister lines or between a recurrent parent and its SBWs. Early flowering was a common feature among lines with broad adaptation and/or high yield in the northern Australian wheatbelt, while yields in the southern region did not show any association with the maturity type. Lines with high yields in the southern and northern regions had cooler canopies during flowering and early grain filling. Among the SBWs with Australian genetic backgrounds, lines best adapted to CIANO were tall (>100 cm), with a slightly higher ground cover. These lines also displayed a higher concentration of water-soluble carbohydrates in the stem at flowering, which was negatively correlated with stem number per unit area when evaluated in southern Australia (Horsham). Possible reasons for these patterns are discussed. Selection for yield at CIANO did not specifically identify the lines best adapted to northern Australia, although they were not the most poorly adapted either. In addition, groups of lines with specific adaptation to the south would not have been selected by choosing the highest yielding lines at CIANO. These findings suggest that selection at CIMMYT for Australian environments may be improved by either trait based selection or yield data combined with trait information. Flowering date, canopy temperature around flowering, tiller density, and water-soluble carbohydrate concentration in the stem at flowering seem likely candidates.

The effect of habitat and environmental history on otolith chemistry of barramundi Lates calcarifer in estuarine populations of a regulated tropical river

We examine the microchemistry of otoliths of cohorts of a fished shed population of the large catadromous fish, barramundi Lates calcarifer from the estuary of a large tropical river. Barramundi from the estuary of the large, heavily regulated Fitzroy River, north eastern Australia were analysed by making transects of 87Sr/86Sr isotope and trace metal/Ca ratios from the core to the outer edge. Firstly, we examined the Sr/Ca, Ba/Ca, Mg/Ca and Mn/Ca and 87Sr/86Sr isotope ratios in otoliths of barramundi tagged in either freshwater or estuarine habitats that were caught by the commercial fishery in the estuary. We used 87Sr/86Sr isotope ratios to identify periods of freshwater residency and assess whether trace metal/Ca ratios varied between habitats. Only Sr/Ca consistently varied between known periods of estuarine or freshwater residency. The relationships between trace metal/Ca and river flow, salinity, temperature were examined in fish tagged and recaptured in the estuary. We found weak and inconsistent patterns in relationships between these variables in the majority of fish. These results suggest that both individual movement history within the estuary and the scale of environmental monitoring were reducing our ability to detect any patterns. Finally, we examined fish in the estuary from two dominant age cohorts (4 and 7 yr old) before and after a large flood in 2003 to ascertain if the flood had enabled fish from freshwater habitats to migrate to the estuary. There was no difference in the proportion of fish in the estuary that had accessed freshwater after the flood. Instead, we found that larger individuals with each age cohort were more likely to have spent a period in freshwater. This highlights the need to maintain freshwater flows in rivers. About half the fish examined had accessed freshwater habitats before capture. Of these, all had spent at least their first two months in marine salinity waters before entering freshwater and some did not enter freshwater until four years of age. This contrasts with the results of several previous studies in other parts of the range that found that access to freshwater swamps by larval barramundi was important for enhanced population productivity and recruitment.

Marine resources, biophysical processes, and environmental management of a tropical shelf seaway: Torres Strait, Australia-Introduction to the special issue

This special issue of Continental Shelf Research contains 20 papers giving research results produced as part of Australia's Torres Strait Co-operative Research Centre (CRC) Program, which was funded over a three-year period during 2003-2006. Marine biophysical, fisheries, socioeconomic-cultural and extension research in the Torres Strait region of northeastern Australia was carried out to meet three aims: 1) support the sustainable development of marine resources and minimize impacts of resource use in Torres Strait; 2) enhance the conservation of the marine environment and the social, cultural and economic well being of all stakeholders, particularly the Torres Strait peoples; and 3) contribute to effective policy formulation and management decision making. Subjects covered, including commercial and traditional fisheries management, impacts of anthropogenic sediment inputs on seagrass meadows and communication of science results to local communities, have broad applications to other similar environments.

Ruminococcus bromii, identification and isolation as a dominant community member in the rumen of cattle fed a barley diet

Aims: To identify dominant bacteria in grain (barley)-fed cattle for isolation and future use to increase the efficiency of starch utilization in these cattle. Methods and Results: Total DNA was extracted from samples of the rumen contents from eight steers fed a barley diet for 9 and 14 days. Bacterial profiles were obtained using denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified V2/V3 region of the 16S rRNA genes from total bacterial DNA. Apparently dominant bands were excised and cloned, and the clone insert sequence was determined. One of the most common and dominant bacteria present was identified as Ruminococcus bromii. This species was subsequently isolated using traditional culture-based techniques and its dominance in the grain-fed cattle was confirmed using a real-time Taq nuclease assay (TNA) designed for this purpose. In some animals, the population of R. bromii reached densities above 1010R. bromii cell equivalents per ml or approximately 10% of the total bacterial population. Conclusions: Ruminococcus bromii is a dominant bacterial population in the rumen of cattle fed a barley-based diet. Significance and Impact of the Study: Ruminococcus bromii YE282 may be useful as a probiotic inoculant to increase the efficiency of starch utilization in barley-fed cattle. The combination of DGGE and real-time TNA has been an effective process for identifying and targeting for isolation, dominant bacteria in a complex ecosystem.

Modelling environmental effects on phenology and canopy development of diverse sorghum genotypes

The ability to predict phenology and canopy development is critical in crop models used for simulating likely consequences of alternative crop management and cultivar choice strategies. Here we quantify and contrast the temperature and photoperiod responses for phenology and canopy development of a diverse range of elite Indian and Australian sorghum genotypes (hybrid and landrace). Detailed field experiments were undertaken in Australia and India using a range of genotypes, sowing dates, and photoperiod extension treatments. Measurements of timing of developmental stages and leaf appearance were taken. The generality of photo-thermal approaches to modelling phenological and canopy development was tested. Environmental and genotypic effects on rate of progression from emergence to floral initiation (E-FI) were explained well using a multiplicative model, which combined the intrinsic development rate (Ropt), with responses to temperature and photoperiod. Differences in Ropt and extent of the photoperiod response explained most genotypic effects. Average leaf initiation rate (LIR), leaf appearance rate and duration of the phase from anthesis to physiological maturity differed among genotypes. The association of total leaf number (TLN) with photoperiod found for all genotypes could not be fully explained by effects on development and LIRs. While a putative effect of photoperiod on LIR would explain the observations, other possible confounding factors, such as air-soil temperature differential and the nature of model structure were considered and discussed. This study found a generally robust predictive capacity of photo-thermal development models across diverse ranges of both genotypes and environments. Hence, they remain the most appropriate models for simulation analysis of genotype-by-management scenarios in environments varying broadly in temperature and photoperiod.

Effects of environmental heat on conception rates in lactating dairy cows: Critical periods of exposure

Environmental heat can reduce conception rates (the proportion of services that result in pregnancy) in lactating dairy cows. The study objectives were to identify periods of exposure relative to the service date in which environmental heat is most closely associated with conception rates, and to assess whether the total time cows are exposed to high environmental heat within each 24-h period is more closely associated with conception rates than is the maximum environmental heat for each 24-h period. A retrospective observational study was conducted in 25 predominantly Holstein-Friesian commercial dairy herds located in Australia. Associations between weather and conception rates were assessed using 16,878 services performed over a 21-mo period. Services were classified as successful based on rectal palpation. Two measures of heat load were defined for each 24-h period: the maximum temperature-humidity index (THI) for the period, and the number of hours in the 24-h period when the THI was >72. Conception rates were reduced when cows were exposed to a high heat load from the day of service to 6 d after service, and in wk -1. Heat loads in wk -3 to -5 were also associated with reduced conception rates. Thus, management interventions to ameliorate the effects of heat load on conception rates should be implemented at least 5 wk before anticipated service and should continue until at least 1 wk after service. High autocorrelations existed between successive daily values in both measures, and associations between day of heat load relative to service day and conception rates differed substantially when ridge regression was used to account for this autocorrelation. This indicates that when assessing the effects of heat load on conception rates, the autocorrelation in heat load between days should be accounted for in analyses. The results suggest that either weekly averages or totals summarizing the daily heat load are adequate to describe heat load when assessing effects on conception rates in lactating dairy cows.