Does reduced usage of antibiotics in livestock production mitigate the spread of antibiotic resistance in soil, earthworm guts, and the phyllosphere?

The overuse of antibiotics in animal husbandry is widespread and believed to significantly contribute to the selection of antibiotic resistance genes (ARGs) in animals. Thus, there is a global drive to reduce antibiotic use in the agricultural sector. However, it has not been established whether a reduction in the use of antibiotics in livestock production would be effective in reducing the spread of ARGs. A microcosm approach was used to determine how the addition of manure with either reduced antibiotic levels or with typical antibiotic levels could affect the spread of antibiotic resistance genes between soil, earthworms and the phyllosphere. When compared to the control soil, earthworm and phyllosphere samples had the greater increase in ARG abundance in conventional manure treatments (P < 0.05). Reduced antibiotic manure also enriched the abundance of ARGs in the phyllosphere and soil but not earthworm guts when compared to the control (P < 0.05). In both soil and earthworm guts, the enrichment of ARGs was lower in reduced antibiotic manure than in conventional manure. This study has identified bacterial transfer through the soil-earthworm-phyllosphere system as a potential means to spread ARGs between habitats after fertilization with livestock derived manures

Ascertaining the nature and timing of mire degradation : using palaeoecology to assist future conservation management in Northern England

This study was initiated under contract VT0419 from English Nature (Natural England) to CECQR. In 2015, additional funding was provided by the Yorkshire Peat Partnership, for 210Pb dating, to refine age–depth estimates. We thank the following for fieldwork co-ordination, logistics or field assistance: Miriam Baynes, Mike Sutcliffe, Martin Furness, Gez Marshall, Paul Duncan, Tim Page, Andrew Windrum, Craig Sandham and Jackie Smith. We thank the Editor and two anonymous reviewers for suggestions for improvements.Peer reviewedPublisher PD

Soil nitrate reducing processes drivers, mechanisms for spatial variation, and significance for nitrous oxide production

The microbial processes of denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are two important nitrate reducing mechanisms in soil, which are responsible for the loss of nitrate (NO− 3 ) and production of the potent greenhouse gas, nitrous oxide (N2O). A number of factors are known to control these processes, including O2 concentrations and moisture content, N, C, pH, and the size and community structure of nitrate reducing organisms responsible for the processes. There is an increasing understanding associated with many of these controls on flux through the nitrogen cycle in soil systems. However, there remains uncertainty about how the nitrate reducing communities are linked to environmental variables and the flux of products from these processes. The high spatial variability of environmental controls and microbial communities across small sub centimeter areas of soil may prove to be critical in determining why an understanding of the links between biotic and abiotic controls has proved elusive. This spatial effect is often overlooked as a driver of nitrate reducing processes. An increased knowledge of the effects of spatial heterogeneity in soil on nitrate reduction processes will be fundamental in understanding the drivers, location, and potential for N2O production from soils

Linking biogeochemical variability and microbial community structure in forest soils: Abies pinsapo-fir forests as a study case

Tradicionalmente, los estudios biogeoquímicos consideraban el suelo como una "caja negra", conectada con el resto del ecosistema mediante flujos de entrada y de salida de nutrientes, en la que éstos experimentan transformaciones mediadas en gran parte por microorganismos desconocidos. El desarrollo de nuevas técnicas moleculares ha incrementado el conocimiento sobre las comunidades microbianas del suelo. Estudios recientes han puesto de manifiesto correspondencias entre la variabilidad de la disponibilidad de nutrientes y la estructura de la comunidad microbiana en suelos forestales. A este respecto, hemos empleado bosques de Abies pinsapo como modelo ya que muestran una gran variabilidad biogeoquímica asociada a diferencias litológicas y sucesionales, mientras que la estacionalidad mediterránea provoca una alta variabilidad intra anual en la disponibilidad de nutrientes. Además, existen pinsapares sujetos a limitación o a saturación de nitrógeno. Mediante análisis del polimorfismo de longitud de fragmentos de restricción terminal (T-RFLP), hemos puesto de manifiesto que los patrones de variabilidad biogeoquímica descritos en los pinsapares se corresponden con cambios en la composición y estructura de sus comunidades bacterianas.Traditionally, soil has been considered as a "black box", connected to the rest of the ecosystem by input and output fluxes, where nutrients are transformed mainly through processes controlled by unknown microorganisms. The development of new molecular techniques has increased our knowledge about microbiological soil communities. Recent studies reveal that change in nutrient availability in forest soils is associated with shifts in the soil community structure. We have used Abies pinsapo fir forests as a model case to this regard, since they show a marked among-sites biogeochemical variability due to changes in lithology and successional status, while Mediterranean seasonality determines high intra-year variability in nutrients availability. Moreover, A. pinsapo forests that are either N-limited or N-saturated do exist. Using terminal restriction fragments length polymorphism (T-RFLP) analysis we have demonstrated that these patterns of biogeochemical variability are associated with shifts in the structure of the soil bacterial communities

The effects of arbuscular mycorrhizal fungal colonisation on nutrient status, growth, productivity, and canker resistance of apple (Malus pumila)

We assess whether arbuscular mycorrhizal fungi (AMF) improve growth, nutritional status, phenology, flower and fruit production, and disease resistance in woody perennial crops using apple (Malus pumila) as a study system. In a fully factorial experiment, young trees were grown for 3 years with or without AMF (Funneliformis mosseae and Rhizophagus irregularis), and with industrial standard fertiliser applications or restricted fertiliser (10% of standard). We use two commercial scions (Dabinett and Michelin) and rootstocks (MM111 and MM106). Industrial standard fertiliser applications reduced AMF colonisation and root biomass, potentially increasing drought sensitivity. Mycorrhizal status was influenced by above ground genotypes (scion type) but not rootstocks, indicating strong interactions between above and below ground plant tissue. The AMF inoculation significantly increased resistance to Neonectria ditissima, a globally economically significant fungal pathogen of apple orchards, but did not consistently alter leaf nutrients, growth, phenology or fruit and flower production. This study significantly advances understanding of AMF benefits to woody perennial crops, especially increased disease resistance which we show is not due to improved tree nutrition or drought alleviation. Breeding programmes and standard management practises can limit the potential for these benefits

Variation in mycorrhizal growth response among a spring wheat mapping population shows potential to breed for symbiotic benefit.

Funder: N8 Agrifood SchemeAll cereal crops engage in arbuscular mycorrhizal symbioses which can have profound, but sometimes deleterious, effects on plant nutrient acquisition and growth. The mechanisms underlying variable mycorrhizal responsiveness in cereals are not well characterised or understood. Adapting crops to realise mycorrhizal benefits could reduce fertiliser requirements and improve crop nutrition where fertiliser is unavailable. We conducted a phenotype screen in wheat (Triticum aestivum L.), using 99 lines of an Avalon × Cadenza doubled-haploid mapping population. Plants were grown with or without a mixed inoculum containing 5 species of arbuscular mycorrhizal fungi. Plant growth, nutrition and mycorrhizal colonisation were quantified. Plant growth response to inoculation was remarkably varied among lines, ranging from more than 30% decrease to 80% increase in shoot biomass. Mycorrhizal plants did not suffer decreasing shoot phosphorus concentration with increasing biomass as observed in their non-mycorrhizal counterparts. The extent to which mycorrhizal inoculation was beneficial for individual lines was negatively correlated with shoot biomass in the non-mycorrhizal state but was not correlated with the extent of mycorrhizal colonisation of roots. Highly variable mycorrhizal responsiveness among closely related wheat lines and the identification of several QTL for these traits suggests the potential to breed for improved crop-mycorrhizal symbiosis

Investigational new drug enabling angiotensin oral-delivery studies to attenuate pulmonary hypertension

Pulmonary arterial hypertension (PAH) is a deadly and uncurable disease characterized by remodeling of the pulmonary vasculature and increased pulmonary artery pressure. Angiotensin Converting Enzyme 2 (ACE2) and its product, angiotensin-(1-7) [ANG-(1-7)] were expressed in lettuce chloroplasts to facilitate affordable oral drug delivery. Lyophilized lettuce cells were stable up to 28 months at ambient temperature with proper folding, assembly of CTB-ACE2/ANG-(1-7) and functionality. When the antibiotic resistance gene was removed, Ang1-7 expression was stable in subsequent generations in marker-free transplastomic lines. Oral gavage of monocrotaline-induced PAH rats resulted in dose-dependent delivery of ANG-(1-7) and ACE2 in plasma/tissues and PAH development was attenuated with decreases in right ventricular (RV) hypertrophy, RV systolic pressure, total pulmonary resistance and pulmonary artery remodeling. Such attenuation correlated well with alterations in the transcription of Ang-(1-7) receptor MAS and angiotensin II receptor AGTRI as well as IL-1β and TGF-β1. Toxicology studies showed that both male and female rats tolerated ~10-fold ACE2/ANG-(1-7) higher than efficacy dose. Plant cell wall degrading enzymes enhanced plasma levels of orally delivered protein drug bioencapsulated within plant cells. Efficient attenuation of PAH with no toxicity augurs well for clinical advancement of the first oral protein therapy to prevent/treat underlying pathology for this disease

Compound driven differences in N2 and N2O emission from soil; the role of substrate use efficiency and the microbial community

Organic C is an important control on the process of denitrification, a process that can result in the production and reduction of the potent greenhouse gas nitrous oxide (N2O). This study identified the influence of different low molecular weight C (LMW-C) compounds on the production of nitrous oxide (N2O) and dinitrogen (N2) and the associated role of the size and structure of the microbial community. We examined this following application of glucose, glutamine or citric acid (250 mg C kg−1 dry soil) and 15N-KNO3 (100 mg N kg−1 dry soil) to a sandy loam soil and measured the production of N2 and N2O by denitrifiers using 15N labeling techniques, changes in the bacterial community as measured by T-RFLP on 16SrDNA fragments and changes in the gene copy number of 16SrDNA, nirK, nirS and nosZ over 144 h. Addition of glucose, citric acid and glutamine all increased emissions of 15N-N2 above that found in the control (P < 0.05) while the addition of glucose and glutamine resulted in higher emissions of 14+15N-N2O (P < 0.001) than the addition of citric acid, resulting in a lower 15N-N2O to 15N-N2 ratio in the citric acid treatment. The 16SrDNA gene copy number increased after addition of citric acid and glutamine, whilst 16SrDNA showed significant shifts in community composition in all C treatments although over different time periods. The gene copy number of nosZ only significantly increased at 120 h in the glutamine treatment (P < 0.05) and nirS at 120 h in the citric acid and glutamine treatments (P < 0.05). This suggests that where C is added as a single input, differences in N2 and N2O emissions between LMW-C compounds were not caused by selection for denitrifiers but likely driven by differences in substrate use efficiency and subsequent differences in C partitioning between growth and respiration. The differing influence of the three selected C compounds on denitrification indicates the potential for lowering net N2O emissions through regulation of C compound availability

Mineral and organic fertilization alters the microbiome of a soil nematode Dorylaimus stagnalis and its resistome

Although the effects of fertilization on the abundance and diversity of soil nematodes have been widely studied, the impact of fertilization on soil nematode microbiomes remains largely unknown. Here, we investigated how different fertilizers: no fertilizer, mineral fertilizer, clean slurry (pig manure with a reduced antibiotic burden) and dirty slurry (pig manure with antibiotics) affect the microbiome of a dominant soil nematode and its associated antibiotic resistance genes (ARGs). The results of 16S rRNA gene high throughput sequencing showed that the microbiome of the soil nematode Dorylaimus stagnalis is diverse (Shannon index: 9.95) and dominated by Proteobacteria (40.3%). Application of mineral fertilizers significantly reduced the diversity of the nematode microbiome (by 28.2%; P < 0.05) but increased the abundance of Proteobacteria (by 70.1%; P = 0.001). Microbial community analysis, using a null hypothesis model, indicated that microbiomes associated with the nematode are not neutrally assembled. Organic fertilizers also altered the diversity of the nematode microbiome, but had no impact on its composition as illustrated by principal coordinates analysis (PCoA). Interestingly, although no change of total ARGs was observed in the nematode microbiome and no significant relationship existed between nematode microbiome and resistome, the abundance of 48 out of a total of 75 ARGs was enriched in the organic fertilizer treatments. Thus, the data suggests that ARGs in the nematode microbiome still had a risk of horizontal gene transfer under fertilization and nematodes might be a potential refuge for ARGs

Testing the activitystat hypothesis: a randomised controlled trial protocol

Background: The activitystat hypothesis proposes that when physical activity or energy expenditure is increased or decreased in one domain, there will be a compensatory change in another domain to maintain an overall, stable level of physical activity or energy expenditure. To date, there has been no experimental study primarily designed to test the activitystat hypothesis in adults. The aim of this trial is to determine the effect of two different imposed exercise loads on total daily energy expenditure and physical activity levels. Methods. This study will be a randomised, multi-arm, parallel controlled trial. Insufficiently active adults (as determined by the Active Australia survey) aged 18-60 years old will be recruited for this study (n=146). Participants must also satisfy the Sports Medicine Australia Pre-Exercise Screening System and must weigh less than 150 kg. Participants will be randomly assigned to one of three groups using a computer-generated allocation sequence. Participants in the Moderate exercise group will receive an additional 150 minutes of moderate to vigorous physical activity per week for six weeks, and those in the Extensive exercise group will receive an additional 300 minutes of moderate to vigorous physical activity per week for six weeks. Exercise targets will be accumulated through both group and individual exercise sessions monitored by heart rate telemetry. Control participants will not be given any instructions regarding lifestyle. The primary outcome measures are activity energy expenditure (doubly labeled water) and physical activity (accelerometry). Secondary measures will include resting metabolic rate via indirect calorimetry, use of time, maximal oxygen consumption and several anthropometric and physiological measures. Outcome measures will be conducted at baseline (zero weeks), mid- and end-intervention (three and six weeks) with three (12 weeks) and six month (24 week) follow-up. All assessors will be blinded to group allocation. Discussion. This protocol has been specifically designed to test the activitystat hypothesis while taking into account the key conceptual and methodological considerations of testing a biologically regulated homeostatic feedback loop. Results of this study will be an important addition to the growing literature and debate concerning the possible existence of an activitystat. Trial registration. Australian New Zealand Clinical Trials Registry ACTRN12610000248066