Application of monoclonal antibodies in quantifying fungal growth dynamics during aerobic spoilage of silage

Proliferation of filamentous fungi following ingress of oxygen to silage is an important cause of dry matter losses, resulting in significant waste. In addition, the production of mycotoxins by some filamentous fungi pose a risk to animal health through mycotoxicosis. Quantitative assessment of fungal growth in silage, through measurement of ergosterol content, colony forming units or temperature increase are limiting in representing fungal growth dynamics during aerobic spoilage due to being deficient in either representing fungal biomass or being able to identify specific genera. Here, we conducted a controlled environment aerobic exposure experiment to test the efficacy of a monoclonal antibody-based enzyme linked immunosorbent assay (ELISA) to detect the proliferation of fungal biomass in six silage samples. We compared this to temperature which has been traditionally deployed in such experiments and on-farm to detect aerobic deterioration. In addition, we quantified ergosterol, a second marker of fungal biomass. At 8 d post aerobic exposure, the ergosterol and ELISA methods indicated an increase in fungal biomass in one of the samples with a temperature increase observed after 16 d. A comparison of the methods with Pearson’s correlation coefficient showed a positive association between temperature and ergosterol and both markers of fungal biomass. This work indicates that the technology has potential to be used as an indicator of microbial degradation in preserved forage. Consequently, if developed as an on farm technique this could inform forage management decisions made by farmers, with the goal of decreasing dry matter losses, improving resource and nutrient efficiency and reducing risks to animal health

The stable oxygen isotope ratio of resin extractable phosphate derived from fresh cattle faeces

Phosphorus losses from agriculture pose an environmental threat to watercourses. A new approach using the stable oxygen isotope ratio of oxygen in phosphate (δ18OPO4 value) may help elucidate some phosphorus sources and cycling. Accurately determined and isotopically distinct source values are essential for this process. The δ18OPO4 values of animal wastes have, up to now, received little attention. Methods Phosphate (PO4) was extracted from cattle faeces using anion resins and the contribution of microbial PO4 was assessed. The δ18OPO4 value of the extracted PO4 was measured by precipitating silver phosphate and subsequent analysis on a thermal conversion elemental analyser at 1400°C, with the resultant carbon monoxide being mixed with a helium carrier gas passed through a GC column into a mass spectrometer. Faecal water oxygen isotope ratios (δ18OH2O values) were determined on a dual-inlet mass spectrometer through a process of headspace carbon dioxide equilibration with water samples. Results Microbiological results indicated that much of extracted PO4 was not derived directly from the gut fauna lysed during the extraction of PO4 from the faeces. Assuming that the faecal δ18OH2O values represented cattle body water, the predicted pyrophosphatase equilibrium δ18OPO4 (Eδ18OPO4) values ranged between +17.9 and +19.9‰, while using groundwater δ18OH2O values gave a range of +13.1 to +14.0‰. The faecal δ18OPO4 values ranged between +13.2 and +15.3‰. Conclusions The fresh faecal δ18OPO4 values were equivalent to those reported elsewhere for agricultural animal slurry. However, they were different from the Eδ18OPO4 value calculated from the faecal δ18OH2O value. Our results indicate that slurry PO4 is, in the main, derived from animal faeces although an explanation for the observed value range could not be determined

Response of soil health indicators to dung, urine and mineral fertilizer application in temperate pastures

Healthy soils are key to sustainability and food security. In temperate grasslands, not many studies have focused on soil health comparisons between contrasting pasture systems under different management strategies and treatment applications (e.g. manures and inorganic fertilisers). The aim of this study was to assess the responses of soil health indicators to dung, urine and inorganic N fertiliser in three temperate swards: permanent pasture not ploughed for at least 20 years (PP), high sugar ryegrass with white clover targeted at 30% coverage reseeded in 2013 (WC), and high sugar ryegrass reseeded in 2014 (HG). This study was conducted on the North Wyke Farm Platform (UK) from April 2017 to October 2017. Soil health indicators including soil organic carbon (SOC, measured by loss of ignition and elemental analyser), dissolved organic carbon (DOC), total nitrogen (TN), C:N ratio, soil C and N bulk isotopes, pH, bulk density (BD), aggregate stability, ergosterol concentration (as a proxy for fungi biomass), and earthworms (abundance, mass and density) were measured and analysed before and after application of dung and N fertilizer, urine and N fertiliser, and only N fertiliser. The highest SOC, TN, DOC, ergosterol concentration and earthworms as well as the lowest BD were found in PP, likely due to the lack of ploughing. Differences among treatments were observed due to the application of dung, resulting in an improvement in chemical indicators of soil health after 50 days of its application. Ergosterol concentration was significantly higher before treatment applications than at the end of the experiment. No changes were detected in BD and aggregate stability after treatment applications. We conclude that not enough time had passed for the soil to recover after the ploughing and reseeding of the permanent pasture, independently of the sward composition (HG or WC). Our results highlight the strong influence of the soil management legacy in temperate pasture and the positive effects of dung application on soil health over the short term. In addition, we point out the relevance of using standardised methods to report soil health indicators and some methodological limitations

Elucidating three-way interactions between soil, pasture and animals that regulate nitrous oxide emissions from temperate grazing systems

Pasture-based livestock farming contributes considerably to global emissions of nitrous oxide (N2O), a powerful greenhouse gas approximately 265 times more potent than carbon dioxide. Traditionally, the estimation of N2O emissions from grasslands is carried out by means of plot-scale experiments, where externally sourced animal excreta are applied to soils to simulate grazing conditions. This approach, however, fails to account for the impact of different sward types on the composition of excreta and thus the functionality of soil microbiomes, creating unrealistic situations that are seldom observed under commercial agriculture. Using three farming systems employing contrasting pasture management strategies at the North Wyke Farm Platform, an instrumented ruminant grazing trial in Devon, UK, this study measured N2O emissions from soils treated with cattle urine and dung collected within each system as well as standard synthetic urine shared across all systems, and compared them against two forms of controls with and without inorganic nitrogen fertiliser applications. Soil microbial activity was regularly monitored through gene abundance to evaluate interactions between sward types, soil amendments, soil microbiomes and, ultimately, N2O production. Across all systems, N2O emissions attributable to cattle urine and standard synthetic urine were found to be inconsistent with one another due to discrepancy in nitrogen content. Despite previous findings that grasses with elevated levels of water-soluble carbohydrates tend to generate lower levels of N2O, the soil under high sugar grass monoculture in this study recorded higher emissions when receiving excreta from cattle fed the same grass. Combined together, our results demonstrate the importance of evaluating environmental impacts of agriculture at a system scale, so that the feedback mechanisms linking soil, pasture, animals and microbiomes are appropriately considered

Space Division Multiplexing in Optical Fibres

Optical communications technology has made enormous and steady progress for several decades, providing the key resource in our increasingly information-driven society and economy. Much of this progress has been in finding innovative ways to increase the data carrying capacity of a single optical fibre. In this search, researchers have explored (and close to maximally exploited) every available degree of freedom, and even commercial systems now utilize multiplexing in time, wavelength, polarization, and phase to speed more information through the fibre infrastructure. Conspicuously, one potentially enormous source of improvement has however been left untapped in these systems: fibres can easily support hundreds of spatial modes, but today's commercial systems (single-mode or multi-mode) make no attempt to use these as parallel channels for independent signals.Comment: to appear in Nature Photonic

Development and validation of a field microphone-in-real-ear approach for measuring hearing protector attenuation

Numerous studies have shown that the reliability of using laboratory measurements to predict individual or even group hearing protector attenuation for occupationally exposed workers is quite poor. This makes it difficult to properly assign hearing protectors when one wishes to closely match attenuation to actual exposure. An alternative is the use of field-measurement methods, a number of which have been proposed and are beginning to be implemented. We examine one of those methods, namely the field microphone-in-real-ear (F-MIRE) approach in which a dual-element microphone probe is used to measure noise reduction by quickly sampling the difference in noise levels outside and under an earplug, with appropriate adjustments to predict real-ear attenuation at threshold (REAT). We report on experiments that validate the ability of one commercially available F-MIRE device to predict the REAT of an earplug fitted identically for two tests. Results are reported on a representative roll-down foam earplug, stemmed-style pod plug, and pre-molded earplug, demonstrating that the 95% confidence level of the Personal Attenuation Rating (PAR) as a function of the number of fits varies from ±4.4 dB to ±6.3 dB, depending on the plug type, which can be reduced to ±3.1 dB to ±4.5 dB with a single repeat measurement. The added measurement improves precision substantially. However, the largest portion of the error is due to the user′s fitting variability and not the uncertainty of the measurement system. Further we evaluated the inherent uncertainty of F-MIRE vs. the putative "gold standard" REAT procedures finding, that F-MIRE measurement uncertainty is less than one-half that of REAT at most test frequencies. An American National Standards Institute (ANSI) working group (S12/WG11) is currently involved in developing methods similar to those in this paper so that procedures for evaluating and reporting uncertainty on all types of field attenuation measurement systems can be standardized. We conclude that the hearing conservationist now has available a portable, convenient, quick, and easy-to-use system that can improve training and motivation of employees, assign hearing protection devices based on noise exposures, and address other management and compliance issues

Identifying risk factors for exposure to culturable allergenic moulds in energy efficient homes by using highly specific monoclonal antibodies

The aim of this study was to determine the accuracy of monoclonal antibodies (mAbs) in identifying culturable allergenic fungi present in visible mould growth in energy efficient homes, and to identify risk factors for exposure to these known allergenic fungi. Swabs were taken from fungal contaminated surfaces and culturable yeasts and moulds isolated by using mycological culture. Soluble antigens from cultures were tested by ELISA using mAbs specific to the culturable allergenic fungi Aspergillus and Penicillium spp., Ulocladium, Alternaria, and Epicoccum spp., Cladosporium spp., Fusarium spp., and Trichoderma spp. Diagnostic accuracies of the ELISA tests were determined by sequencing of the internally transcribed spacer 1 (ITS1)-5.8S-ITS2-encoding regions of recovered fungi following ELISA. There was 100% concordance between the two methods, with ELISAs providing genus-level identity and ITS sequencing providing species-level identities (210 out of 210 tested). Species of Aspergillus/Penicillium, Cladosporium, Ulocladium/Alternaria/Epicoccum, Fusarium and Trichoderma were detected in 82% of the samples. The presence of condensation was associated with an increased risk of surfaces being contaminated by Aspergillus/Penicillium spp. and Cladosporium spp., whereas moisture within the building fabric (water ingress/rising damp) was only associated with increased risk of Aspergillus/Penicillium spp. Property type and energy efficiency levels were found to moderate the risk of indoor surfaces becoming contaminated with Aspergillus/Penicillium and Cladosporium which in turn was modified by the presence of condensation, water ingress and rising damp, consistent with previous literature. (C) 2015 Elsevier Inc. All rights reserved