Predicting mortality of phosphine-resistant adults of Sitophilus oryzae (L) (Coleoptera: Curculionidae) in relation to changing phosphine concentration

Adults of a phosphine-resistant strain of Sitophilus oryzae (L) were exposed to constant phosphine concentrations of 0.0035-0.9 mg litre(-1) for periods of between 20 and 168 h at 25 °C, and the effects of time and concentration on mortality were quantified. Adults were also exposed to a series of treatments lasting 48, 72 or 168 h at 25 °C, during which the concentration of phosphine was varied. The aim of this study was to determine whether equations from experiments using constant concentrations could be used to predict the efficacy of changing phosphine concentrations against adults of S oryzae. A probit plane without interaction, in which the logarithms of time (t) and concentration (C) were variables, described the effects of concentration and time on mortality in experiments with constant concentrations. A derived equation of the form C^nt = k gave excellent predictions of toxicity when applied to data from changing concentration experiments. The results suggest that for resistant S oryzae adults there is nothing inherently different between constant and changing concentration regimes, and that data collected from fixed concentrations can be used to develop equations for predicting mortality in fumigations in which phosphine concentration changes. This approach could simplify the prediction of efficacy of typical fumigations in which concentrations tend to rise and then fall over a period of days.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy® or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to 1 infected in 800 samples with pepper but never detecting more than 1 infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Bats: Important reservoir hosts of emerging viruses

Bats (order Chiroptera, suborders Megachiroptera and Microchiroptera) are abundant, diverse, and geographically widespread. These mammals provide us with resources, but their importance is minimized and many of their populations and species are at risk, even threatened or endangered. Some of their characteristics (food choices, colonial or solitary nature, population structure, ability to fly, seasonal migration and daily movement patterns, torpor and hibernation, life span, roosting behaviors, ability to echolocate, virus susceptibility) make them exquisitely suitable hosts of viruses and other disease agents. Bats of certain species are well recognized as being capable of transmitting rabies virus, but recent observations of outbreaks and epidemics of newly recognized human and livestock diseases caused by viruses transmitted by various megachiropteran and microchiropteran bats have drawn attention anew to these remarkable mammals. This paper summarizes information regarding chiropteran characteristics and information regarding 66 viruses that have been isolated from bats. From these summaries, it is clear that we do not know enough about bat biology, that we are doing too little in terms of bat conservation, and that there remain a multitude of questions regarding the role of bats in disease emergence.

Production of microalgal concentrates by flocculation and their assessment as aquaculture feeds

A novel technique was developed for the flocculation of marine microalgae commonly used in aquaculture. The process entailed an adjustment of pH of culture to between 10 and 10.6 using NaOH, followed by addition of a non-ionic polymer Magnafloc LT-25 to a final concentration of 0.5 mg L-1. The ensuing flocculate was harvested, and neutralised giving a final concentration factor of between 200- and 800-fold. This process was successfully applied to harvest cells of Chaetoceros calcitrans, C. muelleri, Thalassiosira pseudonana, Attheya septentrionalis, Nitzschia closterium, Skeletonema sp., Tetraselmis suecica and Rhodomonas salina, with efficiencies >=80%. The process was rapid, simple and inexpensive, and relatively cost neutral with increasing volume (cf. concentration by centrifugation). Harvested material was readily disaggregated to single cell suspensions by dilution in seawater and mild agitation. Microscopic examination of the cells showed them to be indistinguishable from corresponding non-flocculated cells. Chlorophyll analysis of concentrates prepared from cultures of Concentrates of T. pseudonana prepared using pH-induced flocculation gave better growth of juvenile Pacific oysters (Crassostrea gigas) than concentrates prepared by ferric flocculation, or centrifuged concentrates using a cream separator or laboratory centrifuge. In follow up experiments, concentrates prepared from 1000 L Chaetoceros muelleri cultures were effective as supplementary diets to improve the growth of juvenile C. gigas and the scallop Pecten fumatus reared under commercial conditions, though not as effective as the corresponding live algae. The experiments demonstrated a proof-of-concept for a commercial application of concentrates prepared by flocculation, especially for use at a remote nursery without on-site mass-algal culture facilities.

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.

Review: Near infrared spectroscopy of faeces to evaluate the nutrition and physiology of herbivores

Near infrared (NIR) spectroscopy, usually in reflectance mode, has been applied to the analysis of faeces to measure the concentrations of constituents such as total N, fibre, tannins and delta C-13. In addition, an unusual and exciting application of faecal NIR [F.NIR] analyses is to directly predict attributes of the diet of herbivores such as crude protein and fibre contents, proportions of plant species and morphological components, diet digestibility and voluntary DM intake. This is an unusual application of NIR spectroscopy insofar as the spectral measurements are made, not on the material of interest [i.e. the diet), but on a derived material (i.e. faeces). Predictions of diet attributes from faecal spectra clearly depend on there being sufficient NIR spectral information in the diet residues present in faeces to describe the diet, although endogenous components of faeces such as undigested debris of micro-organisms from the rumen and Large intestine and secretions into the gastrointestinal tract wilt also contribute spectral information. Spectra of forage and of faeces derived from the forage are generally similar and the observed differences are principally in the spectral regions associated with constituents of forages known to be of low, or of high, digestibility. Some diet components (for example, ureal which are likely to be entirely digested apparently cannot be predicted from faecal NIR spectra because they cannot contribute to faecal spectra except through modifying the microbial and endogenous components. The errors and robustness of F.NIR calibrations to predict the crude protein concentration and digestibility of the diet of herbivores are generally comparable with those to directly predict the same attributes in forage from NIR spectra of the forage. Some attributes of the animal, such as species, gender, pregnancy status and parasite burden have been successfully discriminated into classes based on their faecal NIR spectra. Such discrimination was likely associated with differences in the diet selected and/or differences in the metabolites excreted in the faeces. NIR spectroscopy of faeces has usually involved scanning dried and ground samples in monochromators in the 400-2500nm or 1100-2500nm ranges. Results satisfactory for the purpose have also been reported for dried and ground faeces scanned using a diode array instrument in the 800-1700nm range and for wet faeces and slurries of excreta scanned with monochromators. Chemometric analysis of faecal spectra has generally used the approaches established for forage analysis. The capacity to predict many attributes of the diet, and some aspects of animal physiology, from NIR spectra of faeces is particularly useful to study the quality and quantity of the diet selected by both domestic and feral grazing herbivores and to enhance production and management of both herbivores and their grazing environment.

Application of a model to assess aflatoxin risk in peanuts.

When exposed to hot (22-35 degrees C) and dry climatic conditions in the field during the final 4-6 weeks of pod filling, peanuts (Arachis hypogaea L.) can accumulate highly carcinogenic and immuno-suppressing aflatoxins. Forecasting of the risk posed by these conditions can assist in minimizing pre-harvest contamination. A model was therefore developed as part of the Agricultural Production Systems Simulator (APSIM) peanut module, which calculated an aflatoxin risk index (ARI) using four temperature response functions when fractional available soil water was <0.20 and the crop was in the last 0.40 of the pod-filling phase. ARI explained 0.95 (P <= 0.05) of the variation in aflatoxin contamination, which varied from 0 to c. 800 mu g/kg in 17 large-scale sowings in tropical and four sowings in sub-tropical environments carried out in Australia between 13 November and 16 December 2007. ARI also explained 0.96 (P <= 0.01) of the variation in the proportion of aflatoxin-contaminated loads (>15 mu g/kg) of peanuts in the Kingaroy region of Australia during the period between the 1998/99 and 2007/08 seasons. Simulation of ARI using historical climatic data from 1890 to 2007 indicated a three-fold increase in its value since 1980 compared to the entire previous period. The increase was associated with increases in ambient temperature and decreases in rainfall. To facilitate routine monitoring of aflatoxin risk by growers in near real time, a web interface of the model was also developed. The ARI predicted using this interface for eight growers correlated significantly with the level of contamination in crops (r=095, P <= 0.01). These results suggest that ARI simulated by the model is a reliable indicator of aflatoxin contamination that can be used in aflatoxin research as well as a decision-support tool to monitor pre-harvest aflatoxin risk in peanuts.

Seed production of Stylosanthes guyanensis. 1. Growth and development.

The pattern of growth and development of seed crops of stylo (Stylosanthes guyanensis) was derived from measurements made on experimental and commercial crops in north Queensland. The three cultivars Cook, Endeavour, and Schofield differed appreciably only in the timetable of their development. Each had distinct successive phases of vegetative and reproductive development culminating in total annual seed production of 700-800 kg ha-1 from a healthy closed canopy, the main recorded cause of reduced production being the disease Botrytis sp. In a healthy crop of Cook, the peak quantity of standing seed represented almost 90 per cent of the total accountable seed, and the rise to and decline from this peak proceeded at rates of the order of 3-4 per cent per day. It is deduced that, although there appears to be little potential for either increase in overall production or improvement in synchronization or retention characteristics beyond that currently attained by a closed canopy of healthy plants, there is scope for an increase in the efficiency of recovery of standing seed. Maximum recovery will be achieved through attention to choice of time of harvest, presentation of a minimum amount of extraneous vegetation to the harvester, and improvement in harvester separation.

Free Range Hens Use the Range More When the Outdoor Environment Is Enriched.

To evaluate the role of using forage, shade and shelterbelts in attracting birds into the range, three trials were undertaken with free range layers both on a research facility and on commercial farms. Each of the trials on the free range research facility in South Australia used a total of 120 laying hens (Hyline Brown). Birds were housed in an eco-shelter which had 6 internal pens of equal size with a free range area adjoining the shelter. The on-farm trials were undertaken on commercial free range layer farms in the Darling Downs in Southeast Queensland with bird numbers on farms ranging from 2,000-6,800 hens. The first research trial examined the role of shaded areas in the range; the second trial examined the role of forage and the third trial examined the influence of shelterbelts in the range. These treatments were compared to a free range area with no enrichment. Aggressive feather pecking was only observed on a few occasions in all of the trials due to the low bird numbers housed. Enriching the free range environment attracted more birds into the range. Shaded areas were used by 18% of the hens with a tendency (p = 0.07) for more hens to be in the paddock. When forage was provided in paddocks more control birds (55%) were observed in the range in morning than in the afternoon (30%) while for the forage treatments 45% of the birds were in the range both during the morning and afternoon. When shelterbelts were provided there was a significantly (p<0.05) higher % of birds in the range (43% vs. 24%) and greater numbers of birds were observed in areas further away from the poultry house. The results from the on-farm trials mirrored the research trials. Overall 3 times more hens used the shaded areas than the non shaded areas, with slightly more using the shade in the morning than in the afternoon. As the environmental temperature increased the number of birds using the outdoor shade also increased. Overall 17 times more hens used the shelterbelt areas than the control areas, with slightly more using the shelterbelts in the afternoon than in the morning. Approximately 17 times more birds used the forage areas compared to the control area in the corresponding range. There were 8 times more birds using a hay bale enriched area compared to the area with no hay bales. The use of forage sources (including hay bales) were the most successful method on-farm to attract birds into the range followed by shelterbelts and artificial shade. Free range egg farmers are encouraged to provide pasture, shaded areas and shelterbelts to attract birds into the free range.

Relocation of Intensive Agriculture to Northern Australia: The Case of the Rice Industry

Development of new agricultural industries in northern Australia is seen as a way to provide food security in the face of reduced water availability in existing regions in the south. This report aims to identify some of the possible economic consequences of developing a rice industry in the Burdekin region, while there is a reduction of output in the Riverina. Annual rice production in the Riverina peaked at 1.7 M tonnes, but the long-term outlook, given climate change impacts on that region and government water buy-backs, is more likely to be less than 800,000 tonnes. Growers are highly efficient water users by international standards, but the ability to offset an anticipated reduction in water availability through further efficiency gains is limited. In recent years growers in the Riverina have diversified their farms to a greater extent and secondary production systems include beef, sheep and wheat. Production in north Queensland is in its infancy, but a potentially suitable farming system has been developed by including rice within the sugarcane system without competition and in fact contributing to the production of sugar by increasing yields and controlling weeds. The economic outcomes are estimated a large scale, dynamic, computable general equilibrium (CGE) model of the world economy (Tasman Global), scaled down to regional level. CGE models mimic the workings of the economy through a system of interdependent behavioural and accounting equations which are linked to an input-output database. When an economic shock or change is applied to a model, each of the markets adjusts according to the set of behavioural parameters which are underpinned by economic theory. In this study the model is driven by reducing production in the Riverina in accordance with relationships found between water availability and the production of rice and replacement by other crops and by increasing ride production in the Burdekin. Three scenarios were considered: • Scenario 1: Rice is grown using the fallow period between the last ratoon crop of sugarcane and the new planting. In this scenario there is no competition between rice and sugarcane • Scenario 2: Rice displaces sugarcane production • Scenario 3: Rice is grown on additional land and does not compete with sugarcane. Two time periods were used, 2030 and 2070, which are the conventional time points to consider climate change impacts. Under scenario 1, real economic output declines in the Riverina by $45 million in 2030 and by $139 million in 2070. This is only partially offset by the increased real economic output in the Burdekin of $35 million and $131 million respectively.

Electrotransformation of Haemophilus parasuis with in vitro modified DNA based on a novel shuttle vector

The objective of the present study was to establish a valid transformation method of Haemophilus parasuis, the causative agent of Glasser's disease in pigs, using a novel H. parasuis-Escherichia coli shuttle vector. A 4.2 kb endogenous plasmid pYC93 was extracted from an H. parasuis field isolate and completely sequenced. Analysis of pYC93 revealed a region approximately 800 bp showing high homology with the defined replication origin oriV of pLS88, a native plasmid identified in Haemophilus ducreyi. Based on the origin region of pYC93, E. coli cloning vector pBluescript SK(+) and the Tn903 derived kanamycin cassette, a shuttle vector pSHK4 was constructed by overlapping PCR strategy. When electroporation of the 15 H. parasuis serovar reference strains and one clinical isolate SH0165 with pSHK4 was performed, only one of these strains yielded transformants with an efficiency of 8.5 x 10(2) CFUhlg of DNA. Transformation efficiency was notably increased (1.3 x 10(5) CFU/mu g of DNA) with vector DNA reisolated from the homologous transformants. This demonstrated that restriction-modification systems were involved in the barrier to transformation of H. parasuis. By utilizing an in vitro DNA modification method with cell-free extracts of the host H. parasuis strains, 15 out of 16 strains were transformable. The novel shuttle vector pSHK4 and the established electrotransformation method constitute useful tools for the genetic manipulation of H. parasuis to gain a better understanding of the pathogen. (C) 2011 Elsevier B.V. All rights reserved.

Estimating Hydrogen Cyanide in Forage Sorghum (Sorghum bicolor) by Near-Infrared Spectroscopy

Hydrogen cyanide (HCN) is a toxic chemical that can potentially cause mild to severe reactions in animals when grazing forage sorghum. Developing technologies to monitor the level of HCN in the growing crop would benefit graziers, so that they can move cattle into paddocks with acceptable levels of HCN. In this study, we developed near-infrared spectroscopy (MRS) calibrations to estimate HCN in forage sorghum and hay. The full spectral NIRS range (400-2498 nm) was used as well as specific spectral ranges within the full spectral range, i.e., visible (400-750 nm), shortwave (800-1100 nm) and near-infrared (NIR) (1100-2498 nm). Using the full spectrum approach and partial least-squares (PLS), the calibration produced a coefficient of determination (R-2) = 0.838 and standard error of cross-validation (SECV) = 0.040%, while the validation set had a R-2 = 0.824 with a low standard error of prediction (SEP = 0.047%). When using a multiple linear regression (MLR) approach, the best model (NIR spectra) produced a R-2 = 0.847 and standard error of calibration (SEC) = 0.050% and a R-2 = 0.829 and SEP = 0.057% for the validation set. The MLR models built from these spectral regions all used nine wavelengths. Two specific wavelengths 2034 and 2458 nm were of interest, with the former associated with C=O carbonyl stretch and the latter associated with C-N-C stretching. The most accurate PLS and MLR models produced a ratio of standard error of prediction to standard deviation of 3.4 and 3.0, respectively, suggesting that the calibrations could be used for screening breeding material. The results indicated that it should be feasible to develop calibrations using PLS or MLR models for a number of users, including breeding programs to screen for genotypes with low HCN, as well as graziers to monitor crop status to help with grazing efficiency.

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

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

The effect of lycopene on the PI3K/Akt signalling pathway in prostate cancer

Prostate cancer is common in men with very high mortality which is one of leading causes of cancer-related deaths in men. The main treatment approaches for metastasized prostate cancer are androgen deprivation and chemotherapeutic agents. Although there are initial responses to castration, the resistance to the treatment will eventually occur, leading to castration-resistant prostate cancer. The common chemotherapeutic agents for the treatment of prostate cancer are docetaxel and taxane but outcomes of using these drugs have not been satisfactory. Therefore, it is necessary to find better treatment approaches for prostate cancer and to search for compounds that are effective in prostate cancer prevention. Lycopene extracted from tomato and other fruits or plants such as Gac, watermelon, pink grapefruit, pink guava, red carrot and papaya has been shown to be effective on prostate cancer prevention and treatment. The advantage of the application of lycopene for its anti-prostate cancer activity is that lycopene can reach much higher concentration in prostate tissue than other tissues. In this review, the effect of lycopene on PI3K/Akt pathway is summarised, which could be one of major mechanisms for anti-cancer activity of lycopene.

Efficacy and phytotoxicity of different rates of oxadiargyl and pendimethalin in dry-seeded rice (Oryza sativa L.) in Bangladesh

A field study was established to evaluate oxadiargyl and pendimethalin during the wet seasons in Bangladesh in 2012 and 2013. The study evaluated the following treatments: oxadiargyl applied at 80, 120, and 160 g ai ha−1; pendimethalin at 800, 1200, and 1600 g ai ha−1; partial weedy; and weed-free. Rice plant density was greatly affected by weed control treatment. Lower density and lower uniformity of the rice plant stand occurred as a result of increased rates of herbicides. Increased rates of pendimethalin were more toxic than increased rates of oxadiargyl. Both herbicides effectively controlled Digitaria ciliaris, Echinochloa colona, and Phyllanthus niruri; however, they were unable to control Murdannia nudiflora. Oxadiargyl controlled Cyperus rotundus across rates by 31–55%, but pendimethalin was completely ineffective on it, and higher rates of both herbicides had no effect in controlling this weed. Both herbicides at higher rates reduced total weed biomass significantly. Among herbicide treatments, the highest yield (3.7–4.0 t ha−1) was recorded in plots treated with oxadiargyl at 160 g ai ha−1 and the lowest yield (2.4–2.8 t ha−1) was in plots treated with pendimethalin at 1600 g ai ha−1. Results from our study suggest that a higher rate of oxadiargyl can increase yield by suppressing weeds in dry-seeded rice systems. Similar to the results of oxadiargyl, pendimethalin at higher rates also greatly suppressed weeds; however, yield decreased due to phytotoxicity to rice seedlings.